Impacts of Applied Genetics:

Micro-Organisms, Plants, and Animals

April 1981

NTIS order #PB81-206609
Library of Congress Catalog Card Number 81-600046

For sale by the Superintendent of Documents,

U.S. Government Printing Office, Washington, D.C. 20402
Foreword

This report examines the application of classical and molecular genetic technologies to micro-organisms, plants, and animals. Congressional support for an assessment in the field of genetics dates back to 1976 when 30 Representatives requested a

study of recombinant DNA technology. Letters of support for this broader study came

from the then Senate Committee on Human Resources and the House Committee on

Interstate and Foreign Commerce, Subcommittee on Health and the Environment.

Current developments are especially rapid in the application of genetic technologies to micro-organisms; these were studied in three industries: pharmaceutical,

chemical, and food. Classical genetics continue to play the major role in plant and

animal breeding but new genetic techniques are of ever-increasing importance.

This report identifies and discusses a number of issues and options for the Congress, such as:

● Federal Government support of R&D,

● methods of improving the germplasm of farm animal species,

. risks of genetic engineering,

 patenting living organisms, and

● public involvement in decisionmaking.

The Office of Technology Assessment was assisted by an advisory panel of scientists, industrialists, labor representatives, and scholars in the fields of law, economics,

and those concerned with the relationships between science and society. Others contributed in two workshops held during the course of the assessment. The first was to

investigate public perception of the issues in genetics; the second examined genetic

applications to animals. Sixty reviewers drawn from universities, Government, industry, and the law provided helpful comments on draft reports. The Office expresses

sincere appreciation to all those individuals.

An abbreviated copy of the summary of this report (ch. 1) is available free of

charge from the Office of Technology Assessment, U.S. Congress, Washington, D. C.,

20510. In addition, the working papers on the use of genetic technology in human and

in veterinary medicine are available as a separate volume from the National Technical

Information Service.

Director
Impacts of Applied Genetics Advisory

J. E. Legates, Chairman

Dean, School of Agriculture and Life Sciences, North Carolina State University

Ronald E. Cape

Cetus Corp.

Nina V. Fedoroff

Department of Embryology

Carnegie Institution of Washington

June Goodfield

The Rockefeller University

Harold P. Green

Fried, Frank, Harris, Shriver and Kampelman

Halsted R. Holman

Stanford University Medical School

M. Sylvia Krekel

Health and Safety Office

Oil, Chemical, and Atomic Workers

International Union

Elizabeth Kutter

The Evergreen State College

Oliver E. Nelson, Jr.

Laboratory of Genetics

University of Wisconsin

David Pimentel

Department of Entomology

Cornell University

Robert Weaver

Department of Agricultural Economics

Pennsylvania State University

James A. Wright

Pioneer Hi-Bred International

Plant Breeding Division

Norton D. Zinder

The Rockefeller University

iv
Applied Genetics Assessment Staff

Joyce C. Lashof, Assistant Director, OTA

Health and Life Sciences Division

Gretchen Kolsrud, Program Manager

Zsolt Harsanyi, Project Director

Project Staff

Marya Breznay, Administrative Assistant

Lawrence Burton, Analyst

Susan Clymer, Research Assistant

Renee G. Ford, * Technical Editor

Michael Gough, Senior Analyst

Robert Grossman,* Analyst

Richard Hutton, * Editor

Geoffrey M. Karny, Legal Analyst

Major Contractors

Benjamin G. Brackett, University of Pennsylvania

The Genex Corp.

William P. O’Neill, Poly-Planning Services

Plant Resources Institute

Anthony J. Sinskey, Massachusetts Institute of Technology

Aladar A. Szalay, Boyce-Thompson Institute

Virginia Walbot, Washington University

OTA Publishing Staff

John C. Holmes, publishing Officer

John Bergling* Kathie S. Boss Debra M. Datcher

Patricia A. Dyson* Mary Harvey * Joe Henson

OTA contract personnel.

v
Contents

Page

Glossary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. .. .viii. . .

Acronyms and Abbreviations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xii

Chapter l.

Chapter 2.

Chapter 3.

Chapter 4.

Chapter 5.

Chapter 6.

Chapter 7.

Summary: Issues and Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .......00 29

Part I :Biotechnology

Genetic Engineering and the Fermentation Technologies . . . . . . . . . . 49

The Pharmaceutical Industry. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59

The Chemical Industry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85

The Food Processing Industry. . . . . . . . . . . . . . . . . . . . . . . . . .. ....107

The Use of Genetically Engineered Micro-Organisms in the

Environment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ......117

Part II: Agriculture

Chapter 8. The Application of Genetics to Plants . . . . . . . . . . . . . . . . . . . . .. ...137

Chapter 9. Advances in Reproductive Biology and Their Effects on

Animal Improvement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167

Chapter l0.

Chapter ll.

Chapter 12.

Chapter 13.

Appendixes

Part III: Institutions and Society

The Question of Risk. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .......199

Regulation of Genetic Engineering. . . . . . . . . . . . . . . . . . . . . .. ....211

Patenting Living Organisms. . . . . . . . . . . . . . . . . . . . . . ..........237

Genetics and Society . . . . . . . . . . . . . . . . . . . . . ................257

I-A. A Case Study of Acetaminophen Production. . . . . . . . . . . . . . . . . . . . . ...269

I-B. A Timetable for the Commercial Production of Compounds Using

Genetically Engineered Micro-Organisms in Biotechnology . . . . ........275

I-C. Chemical and Biological Processes. . . . . . . . . . . . . . . . . . . . . . ..........292

I-D. The Impact of Genetics on Ethanol-A Case Study . . . . . . .............293

II-A. A Case Study of Wheat. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. ...304

11-B. Genetics and the Forest Products Industry Case Study . . . . . . . .........307

II-C. Animal Fertilization Technologies. . . . . . . . . . . . . . . . . . . . . . ..........309

III-A. History of the Recombinant DNA Debate . . . . . . . . . . . . . . . . . . . . .. ...315

III-B. Constitutional Constraints on Regulation. . . . . . . . . . . . . . . . . . . . . .. ...320

111-C. Information on International Guidelines for Recombinant DNA. . ......322

IV. Planning Workshop Participants, Other Contractors and Contributors, and

Acknowledgments. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...329

vii
Glossary

Aerobic.—Growing only in the presence of oxygen.

Anaerobic. —Growing only in the absence of

oxygen.

Alkaloids.—A group of nitrogen-containing organic

substances found in plants; many are pharmacologically active—e.g., nicotine, caffeine, and

cocaine.

Allele.—Alternate forms of the same gene. For example, the genes responsible for eye color (blue,

brown, green, etc.) are alleles.

Amino acids.—The building blocks of proteins.

There are 20 common amino acids; they are’

joined together in a strictly ordered “string”

which determines the character of each protein.

Antibody.—A protein component of the immune

system in mammals found in the blood.

Antigen.—A large molecule, usually a protein or

carbohydrate, which when introduced in the

body stimulates the production of an antibody

that will react specifically with the antigen.

Aromatic chemical.—An organic compound containing one or more six-membered rings.

Aromatic polymer.–Large molecules consisting

of repeated structural units of aromatic chemicals.

Artificial insemination.—The manual placement

of sperm into the uterus or oviduct.

Bacteriophage (or phage).–A virus that multiplies in bacteria. Bacteriophage lambda is commonly used as a vector in recombinant DNA experiments.

Bioassay .–Determination of the relative strength

of a substance (such as a drug) by comparing its

effect on a test organism with that of a standard

preparation.

Biomass.—Plant and animal material.

Biome.—A community of living organisms in a major ecological region.

Biosynthesis. —The production of a chemical compound by a living organism.

Biotechnology .—The collection of industrial processes that involve the use of biological systems.

For some of these industries, these processes involve the use of genetically engineered microorganisms.

Blastocyst.— An early developmental stage of the

embryo; the fertilized egg undergoes several cell

divisions and forms a hollow ball of cells called

the blastocyst.

Callus.—The cluster of plant cells that results from

tissue culturing a single plant cell.

Carbohydrates.— The family of organic molecules

consisting of simple sugars such as glucose and

sucrose, and sugar chains (polysaccharides) such

as starch and cellulose.

Catalyst.—A substance that enables a chemical

reaction to take place under milder than normal

conditions (e.g., lower temperatures). Biological

catalysts are enzymes; nonbiological catalysts include metallic complexes.

Cell fusion.—The fusing together of two or more

cells to become a single cell.

Cell lysis.—Disruption of the cell membrane allowing the breakdown of the cell and exposure of its

contents to the environment.

Cellulase.—An enzyme that degrades cellulose to

glucose.

Cellulose.–A polysaccharide composed entirely of

several glucose units linked end to end; it constitutes the major part of cell walls in plants.

Chimera.—An individual composed of a mixture of

genetically different cells.

Chloroplast.-The structure in plant cells where

photosynthesis occurs.

Chromosomes.—The thread-like components of a

cell that are composed of DNA and protein. They

contain most of the cell’s DNA.

Clone.—A group of genetically identical cells or

organisms asexually descended from a common

ancestor. All cells in the clone have the same genetic material and are exact copies of the original.

Conjugation.—The one-way transfer of DNA between bacteria in cellular contact.

Crossing-over. —A genetic event that can occur

during celluar replication, which involves the

breakage and reunion of DNA molecules.

Cultivar.—An organism developed and persistent

under cultivat ion.
Cytognetics.-A branch of biology that deals with

the study of heredity and variation by the methods of both cytology (the study of cells) and

genetics.

Cytoplasm.—The protoplasm of a cell, external to

the cell’s nuclear membrane.

Diploid.—A cell with double the basic chromosome

number.

DNA (deoxyribonucleic acid).—The genetic material found in all living organisms. Every inherited characteristic has its origin somewhere in

the code of each individual’s complement of DNA.

DNA vector.—A vehicle for transferring DNA from

one cell to another.

Dominant gene.—A characteristic whose expression prevails over alternative characteristics for a

given trait.

Escherichia coli.-A bacterium that commonly inhabits the human intestine. It is a favorite organism for many microbiological experiments.

Endotoxins.-Complex molecules (lipopolysaccharides) that compose an integral part of the cell

wall, and are released only when the integrity of

the cell is disturbed.

Embryo transfer.—Implantation of an embryo

into the oviduct or uterus.

Enzyme.—A functional protein that catalyzes a

chemical reaction. Enzymes control the rate of

metabolic processes in an organism; they are the

active agents in the fermentation process.

Estrogens.—Female sex hormones.

Estrus (“heat").-The period in which the female

will allow the male to mate her.

Eukaryote.—A higher, compartmentalized cell

characterized by its extensive internal structure

and the presence of a nucleus containing the

DNA. All multicellular organisms are eukaryotic.

The simpler cells, the prokaryotes, have much

less compartmentalization and internal structure; bacteria are prokaryotes.

Exotoxins.—Proteins produced by bacteria that are

able to diffuse out of the cells; generally more potent and specific in their action than endotoxins.

Fermentation.—The biochemical process of converting a raw material such as glucose into a

product such as ethanol.

Fibroblast.—A cell that gives rise to connective

tissues.

Gamete.—A mature reproductive cell.

Gene.–The hereditary unit; a segment of DNA

coding for a specific protein.

Gene expression.—The manifestation of the genetic material of an organism as specific traits.

Genetic drift.—Changes of gene frequency in small

population due to chance preservation or extinction of particular genes.

Genetic code.—The biochemical basis of heredity

consisting of codons (base triplets along the DNA

sequence) that determine the specific amino acid

sequence in proteins and that are the same for all

forms of life studied so far.

Genetic engineering.-A technology used at the

laboratory level to alter the hereditary apparatus

of a living cell so that the cell can produce more

or different chemicals, or perform completely

new functions. These altered cells are then used

in industrial production.

Gene mapping-Determining the relative locations of different genes on a given chromosome.

Genome.—The basic chromosome set of an

organism—the sum total of its genes.

Genotype.—The genetic constitution of an individual or group.

Germplasm.-The total genetic variability available

to an organism, represented by the pool of germ

cells or seed.

Germ cell.—The sex cell of an organism (sperm or

egg, pollen or ovum). It differs from other cells in

that it contains only half the usual number of

chromosomes. Germ cells fuse during fertilization.

Glycopeptides.—Chains of amino acids with attached carbohydrates.

Glycoprotein.—A conjugated protein in which the

nonprotein group is a carbohydrate.

Haploid.—A cell with only one set (half of the usual

number) of chromosomes.

Heterozygous.—When the two genes controlling a

particular trait are different, the organism is

heterozygous for that trait.

Homozygous.— When the two genes controlling a

particular trait are identical for a pair of chromosomes, the organism is said to be homozygous

for that trait.

Hormones.—The “messenger” molecules of the

body that help coordinate the actions of various

tissues; they produce a specific effect on the activity of cells remote from their point of origin.

ix
Hybrid.—A new variety of plant or animal that re- Nif genes.—The genes for nitrogen fixation present

suits from cross-breeding two different existing in certain bacteria.

varieties. Nucleic acid.—A polymer composed of DNA or

Hydrocarbon. —All organic compounds that are RNA subunits.

composed only of carbon and hydrogen. Nucleotides.—The fundamental units of nucleic

Immunoproteins.— All the proteins that are part acids. They consist of one of the four bases—

of the immune system (including antibodies, in- adenine, guanine, cytosine, and thymine (uracil

terferon, and cytokines). in the case of RNA)—and its attached sugar-phosIn vitro. -outside the living organism and in an phate group.

artificial environment. Organic compounds.—Chemical compounds

In vivo.—Within the living organism. based on carbon chains or rings, which contain

hydrogen, and also may contain oxygen, nitroLeukocytes.—The white cells of blood. gen, and various other elements.

Lipids.–Water insoluble biomolecules, such as cel- Parthenogenesis.— Reproduction in animals withlular fats and oils. out male fertilization of the egg.

Lipopolysaccharides. —Complex substances com- Pathogen.—A specific causative agent of disease.

posed of lipids and polysaccharides. Peptide. —Short chain of amino acids.

Lymphoblastoid.– Referring to malignant white

blood cells. pH.–A measure of the acidity or basicity of a solution; on a scale of O (acidic) to 14 (basic): for examLymphokines.–The biologically active soluble factor produced by white blood cells.

Maleic anhydride.—An important organic chemical used in the manufacture of synthetic resins,

in fungicides, in the dyeing of cotton textiles, and

to prevent the oxidation of fats and oils during

storage and rancidity.

Messenger RNA.–Ribonucleic acid molecules that

serve as a guide for protein synthesis.

Metabolism.—The sum of the physical and chemical processes involved in the maintenance of life

and by which energy is made available.

Mitochondria.—Structures in higher cells that

serve as the “powerhouse” for the cell, producing

chemical energy.

Monoclinal antibodies.—Antibodies derived

from a single source or clone of cells which

recognize only one kind of antigen.

Mutants.–Organisms whose visible properties with

respect to some trait differ from the norm of the

population due to mutations in its DNA.

pie, lemon juice has a pH of 2.2 (acidic), water has

a pH of 7.0 (neutral), and a solution of baking

soda has a pH of 8.5 (basic).

Phage.—(See bacteriophage.)

Phenotype. —The visible properties of an organism

that are produced by the interaction of the genotype and the environment.

Plasmid.—Hereditary material that is not part of a

chromosome. Plasmids are circular and self-replicating. Because they are generally small and relatively simple, they are used in recombinant DNA

experiments as acceptors of foreign DNA.

Plastid.–Any specialized organ of the plant cell

other than the nucleus, such as the chloroplast.

Ploidy. —Describes the number of sets of chromosomes present in the organism. For example,

humans are diploid, having two homologous sets

of 23 chromosomes (one set from each parent)

for a total of 46 chromosomes; many plants are

haploid, having only one copy of each chromosome.

Mutation.—Any change that alters the sequence of Polymer.—A long-chain molecule formed from

bases along the DNA, changing the genetic ma- smaller repeating structural units.

terial. Polysaccharide. —A long-chain carbohydrate conMyeloma.—A malignant disease in which tumor taining at least three molecules of simple sugars

cells of the antibody producing system synthesize linked together; examples would include celluexcessive amounts of specific proteins. lose and starch.

n-alkanes.—Straight chain hydrocarbons–the Progestogens. —Hormones involved with ovulamain constituents of petroleum. tion.

x
Prostaglandin.-Refers to a group of naturally occurring, chemically related long-chain fatty acids

that have certain physiological effects (stimulate

contraction of uterine and other smooth muscles,

lower blood pressure, affect action of certain

hormones).

Protein.—A linear polymer of amino acids; proteins

are the products of gene expression and are the

functional and structural components of cells.

Protoplasm.—A cell without a wall.

Protoplasm fusion.—A means of achieving genetic

transformation by joining two protoplasts or joining a protoplasm with any of the components of

another cell.

Recessive gene.—Any gene whose expression is

dependent on the absence of a dominant gene.

Recombinant DNA.–The hybrid DNA produced

by joining pieces of DNA from different sources.

Restriction enzyme.—An enzyme within a bacterium that recognizes and degrades DNA from

foreign organisms, thereby preserving the genetic integrity of the bacterium. In recombinant

DNA experiments, restriction enzymes are used

as tiny biological scissors to cut up foreign DNA

before it is recombined with a vector.

Reverse transcriptase.—An enzyme that can synthesize a single strand of DNA from a messenger

RNA, the reverse of the normal direction of processing genetic information within the cell.

RNA (ribonucleic acid).—In its three forms—messenger RNA, transfer RNA, and ribosomal RNA—

it assists in translating the genetic message of

DNA into the finished protein.

Somatic cell.—One of the cells composing parts of

the body (e.g., tissues,

cell.

Tissue culture.—An in

ing healthy cells from

from skin.

organs) other than- a germ

vitro method of propagattissues, such as fibroblasts

Transduction.—The process by which foreign

DNA becomes incorporated into the genetic complement of the host cell.

Transformation.—The transfer of genetic information by DNA separated from the cell.

Vector.—A transmission agent; a DNA vector is a

self-replicating DNA molecule that transfers a

piece of DNA from one host to another.

Virus.–An infectious agent that requires a host cell

in order for it to replicate. It is composed of

either RNA or DNA wrapped in a protein coat.

Zygote.—A cell formed by the union of two mature

reproductive cells.

xi
Acronyms and Abbreviations

AA

ACS

ACTH

AI

AIPL

APAP

ASM

bbl

bbl/d

BOD5

BBM

bu

CaMV

CCPA

CDC

CERB

DHHS

DHI

DNA

DOD

DOD

DOE

DPAG

EOR

EPA

FDA

FMDV

ftz

ft

FTC

g

gal

GH

ha

HEW

hGH

HYV

— amino acids

— American Cancer Society

— adrenocorticotropic hormone

— artificial insemination

— Animal Improvement Programs

Laboratory

— acetaminophen

— American Society for Microbiology

– barrel(s)

– barrels per day

— 5-day biochemical oxygen demand

— Biological Response Modifier Program

– bushel

— cauliflower mosaic virus

— The Court of Customs and Patent

Appeals

— Center for Disease Control

— Cambridge Experimentation Review

Board

— Department of Health and Human

Services (formerly Health, Education,

and Welfare)

— Dairy Herd Improvement

– deoxyribonucleic acid

— Department of Commerce

— Department of Defense

— Department of Energy

— Dangerous Pathogens Advisory Group

— enhanced oil recovery

— Environmental Protection Agency

— Food and Drug Administration

— foot-and-mouth disease virus

— square foot

– foot

— Federal Trade Commission

– gram

– gallon

– growth hormone

— hectares

— Department of Health, Education, and

Welfare

– human growth hormone

— high-yielding varieties

IBCs — Institutional Biosafety Committees

ICI — Imperial Chemical Industries

IND — Investigational New Drug Application

(FDA)

kg – kilogram

1 — liter

lb – pound

mg — milligram

1% — microgram

pm — micrometer (formerly micron)

MUA — Memorandum of Understanding and

Agreement

NCI — National Cancer Institute

NDA — new drug application (FDA)

NDAB — National Diabetics Advisory Board

NDCHIP — National Cooperative Dairy Herd

Program

NIAID — National Institute of Allergy and

Infectious Diseases

NIAMDD — National Institute of Arthritis,

Metabolism, and Digestive Diseases

NIH – National Institutes of Health

NIOSH — National Institute of Occupational

Safety and Health

NSF — National Science Foundation

OECD — The Organization for Economic

Cooperation and Development

ORDA — Office of Recombinant DNA Activities

PD — predicted difference

pH — unit of measure for acidity/ basicity

ppm – parts per million

R&D — research and development

RAC — Recombinant DNA Advisory Committee

rDNA — recombinant DNA

SCP — single-cell protein

T-DNA — a smaller segment of the the plasmid

Ti — tumor inducing

TSCA — Toxic Substances Control Act
chapter 1

Summary: Issues and Options
Chapter 1

Page

Biotechnology . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

The Pharmaceutical industry. . . . . . . . . . . . . . 4

Findings ... ... ..... . . . . . . . . . . . . . . . . 4

The Chemical Industry. . . . . . . . . . . . . . . . . . . 7

Findings . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

Food processing industry. . . . . . . . . . . . . . . . . 8

Findings . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

The Use of Genetically Engineered MicroOrganisms in the Environment . . . . . . . . . . . 8

Findings . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

Mineral Leaching and Recovery . . . . . . . . . . 9

Enhanced Oil Recovery. . . . . . . . . . . . . . . . . 9

Pollution Control . . . . . . . . . . . . . . . . . . . . . 9

Constraints in Using Genetic Engineering

Technologies in Open Environments. . . . . . . 10

Issue and Options—Biotechnology . . . . . . . . . . 10

Agriculture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

The Applications of Genetics to Plants . . . . . . . 11

Findings . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

New Genetic Technologies for Plant Breeding 12

Constraints on Using Molecular Genetics

for Plant Improvements. . . . . . . . . . . . . . . 13

Genetic Variability, CropVulnerability, and

the Storage of Germplasm . . . . . . . . . . . . . 13

Issues and Options—Plants. . . . . . . . . . . . . . . . 14

Advances in Reproductive Biology and Their

Effects on Animal Improvement . . . . . . . . . . 15

Page

Findings . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

Issue and Options-Animals . . . . . . . . . . . . . . . 17

Institutions and Society. . . . . . . . . . . . . . . . . 18

Regulation of Genetic Engineering . . . . . . . . . . 18

Findings . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

Issue and Options—Regulation . . . . . . . . . . . . . 20

Patenting Living Organisms . . . . . . . . . . . . . . . 22

Findings . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

Issue and Options—Patenting Living Organisms 23

Genetics and Society . . . . . . . . . . . . . . . . . . . . 24

Issues and options—Genetics and Society. .. . 24

Table

Table No. Page

1. Containment Recommended by the National

Institutes of Health... . . . . . . . . . . . . . . . . . . 19

Figures

Figure No. Page

1. Recombinant DNA: The Technique of

Recombining Genes From One Species With

Those From Another.. . . . . . . . . . . . . . . . . . . 5

2. The Product Development Process . . . . . . . . . 6

3. The Way the Reproductive Technologies

Interrelate . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Chapter 1

Summary: Issues and Options

The genetic alteration of plants, animals, and

micro-organisms has been an important part of

agriculture for centuries. It has also been an integral part of the alcoholic beverage industry

since the invention of beer and wine; and for

the past century, a mainstay of segments of the

pharmaceutical and chemical industries.

However, only in the last 20 years have powerful new genetic technologies been developed

that greatly increase the ability to manipulate

the inherited characteristics of plants, animals,

and micro-organisms. One consequence is the

increasing reliance the pharmaceutical and

chemical industries are placing on biotechnology. Micro-organisms are being used to manufacture substances that have previously been

extracted from natural sources. Animal and

plant breeders are using the new techniques to

help clarify basic questions about biological

functions, and to improve the speed and efficiency of the technologies they already use.

Other industries–from food processing and pollution control to mining and oil recovery—are

considering the use of genetic engineering to increase productivity and cut costs.

Genetic technologies will have a broad impact

on the future. They may contribute to filling

some of the most fundamental needs of mankind–from health care to supplies of food and

energy. At the same time, they arouse concerns

about their potential effects on the environment

and the risks to health involved in basic and

applied scientific research and development

(R&D). Because genetic technologies are already

being applied, it is appropriate to begin considering their potential consequences.

Congressional concern with applied genetics

dates back to 1976, when 30 Representatives requested an assessment of recombinant DNA

(rDNA) technology. Support for the broader

study reported here came in letters to the Office

of Technology Assessment from the then Senate

Committee on Human Resources and the House

Committee on Interstate and Foreign Commerce, Subcommittee on Health and the Environment. In addition specific subtopics are of sp

interest to other committees, notably those having jurisdiction over science and technology and

those concerned with patents.

This report describes the potentials and problems of applying the new genetic technologies to

a range of major industries. It emphasizes the

present state of the art because that is what

defines the basis for the future applications. It

then makes some estimates of economic, environmental, and institutional impacts—where,

when, and how some technologies might be applied and what some of the results might be.

The report closes with the possible roles that

Government, industry, and the public might

play in determining the future of applied

genetics.

The term applied genetics, as used in this

report, refers to two groups of technologies:

● Classical genetics—natural mating methods

for the selective breeding of organisms

for desired characteristics–e. g., breeding

cows for increased milk production. The

pool of genes available for selection is comprised of those that cause natural differences among individuals in a population

and those obtained by mutation.

● Molecular genetics includes the technologies

of genetic engineering that involve the

directed manipulation of the genetic material itself. These technologies—such as

rDNA and the chemical synthesis of genes

—can increase the size of the gene pool for

any one organism by making available genetic traits from many different populations. Molecular genetics also includes

technologies in which manipulation occurs

at a level higher than that of the gene—at

the cellular level, e.g., cell fusion and in

vitro fertilization.

Significant applications of molecular genetics

to micro-organisms, such as the efforts to manufacture human insulin, are already underway

in several industries. Most of these applications

3
4 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

depend on fermentation—a technology in which plants and animals, which are biologically more

substances produced by micro-organisms can complex and more difficult to manipulate sucbe obtained in large quantities. Applications to cessfully, will take longer to develop.

Biotechnology

Biotechnology-the use of living organisms or

their components in industrial processes—is

possible because micro-organisms naturally produce countless substances during their lives.

Some of these substances have proved commercially valuable. A number of different industries

have learned to use micro-organisms as natural

factories, cultivating populations of the best

producers under conditions designed to enhance their abilities.

Applied genetics can play a major role in improving the speed, efficiency, and productivity

of these biological systems. It permits the manipulation, or engineering, of the micro-organisms’ genetic material to produce the desired

characteristics. Genetic engineering is not in

itself an industry, but a technique used at the

laboratory level that allows the researcher to

modify the hereditary apparatus of the cell. The

population of altered identical cells that grows

from the first changed micro-organism is, in

turn, used for various industrial processes. (See

figure 1.)

The first major commercial effects of the application of genetic engineering will be in the

pharmaceutical, chemical, and food processing

industries. Potential commercial applications of

value to the mining, oil recovery, and pollution

control industries—which may desire to use manipulated micro-organisms in the open environ -

ment —are stll somewhat speculative.

The pharmaceutical industry

FINDINGS

The pharmaceutical industry has been the

first to take advantage of the potentials of applied molecular genetics. Ultimately, it will

probably benefit more than any other, with the

largest percentage of its products depending on

advances in genetic technologies. Already,

micro-organisms have been engineered to produce human insulin, interferon, growth hormone, urokinase (for the treatment of blood

clots), thymosin-a 1 (for controlling the immune

response), and somatostatin (a brain hormone).

(See figure 2.)

The products most likely to be affected by

genetic engineering in the next 10 to 20 years

are nonprotein compounds like most antibiotics,

and protein compounds such as enzymes and

antibodies, and many hormones and vaccines.

Improvements can be made both in the products and in the processes by which they are produced. Process costs may be lowered and even

entirely new products developed.

The most advanced applications today are in

the field of hormones. While certain hormones

have already proved useful, the testing of

others has been hindered by their scarcity and

high cost. Of 48 human hormones that have

been identified so far as possible candidates for

production by genetically engineered microorganisms, only 10 are used in current medical

practice. The other 38 are not, partly because

they have been available in such limited quantities that tests of their therapeutic value have

not been possible.

Genetic technologies also open up new approaches for vaccine development for such intractable parasitic and viral diseases as amebic

dysentery, trachoma, hepatitis, and malaria. At

present, the vaccine most likely to be produced

is for foot-and-mouth disease in animals. However, should any one of the vaccines for human

diseases become available, the social, economic,

and political consequences of a decrease in morbidity and mortality would be significant. Many

of these diseases are particularly prevalent in

less industrialized countries; the developments

of vaccines for them may profoundly affect the

lives of tens of millions of people.
Ch. 1 Summary: Issues and Options ● 5

Figure 1.— Recombinant DNA: The Technique of Recombining Genes

From One Species With Those From Another

Electron micrograph of the DNA, which is the plasmid SPO1

from Bacillus subtilis. This plasm id which has been

sliced open is used for recombinant DNA research

Donor DNA

in this bacterial host

amount of DNA

SOURCE: Office of Technology Assessment.

Expression

produces

protein

For some pharmaceutical products, biotechnology will compete with chemical synthesis

and extraction from human and animal organs.

Assessing the relative worth of each method

must be done on a case-by-case basis. But for

other products, genetic engineering offers the

only method known that can ensure a plentiful

supply; in some instances, it has no competition.

By making a pharmaceutical available, genetic engineering may have two types of effects:

● Drugs that already have medical promise

Photo credits; Professor F. A. Eiserling, UCLA Molecular Biology Institute

Electron micrograph of Bacillus subtilis in the process of

cell division. The twisted mass in the center of each

daughter cell is the genetic material, DNA

Restriction enzymes recognize certain sites along the DNA

and can chemically cut the DNA at those sites. This makes

it possible to remove selected genes from donor DNA molecules and insert them into plasmid DNA molecules to form

the recombinant DNA. This recombinant DNA can then be

cloned in its bacterial host and large amounts of a desired

protein can be produced.

will be available in ample amounts for clinical testing. Interferon, for example, can be

tested for its efficacy in cancer and viral

therapy, and human growth hormone can

be evaluated for its ability to heal wounds.

● Other pharmacologically active substances

for which no apparent use now exists will

be available in sufficient quantities and at

low enough cost to enable researchers to

explore new uses. As a result, the potential

for totally new therapies exists. Regulatory

proteins, for example, which are an entire

76-565 0 - 81 - 2
6 . Impacts of Applied Genetics—Micro-Organ/ims, Plants, and Animals

Figure 2.-The Product Development Process

2. Tissues

Micro-organisms such as E co/I

I 7. cutting 6. Plasmid

14. Process

development

scale-up

16. Industrial

The development process begins by obtaining DNA either through organic synthesis (1) or derived from biological sources such as tissues

(2). The DNA obtained from one or both sources is tailored to form the basic “gene” (3) which contains the genetic information to “code” for a

desired product, such as human interferon or human insulin. Control signals (4) containing instructions are added to this gene (5). Circular DNA

molecules called plasmids (6) are isolated from micro-organisms such as E. coIi; cut open (7) and spliced back(8) together with genes and control signals to form “recombinant DNA” molecules. These molecules are then introduced into a host cell (9).

Each plasmid is copied many times in a cell (10). Each cell then translates the information contained in these plasmids into the desired product, a process called “expression” (11). Cells divide (12) and pass on to their offspring the same genetic information contained in the parent

cell.

Fermentation of large populations of genetically engineered micro-organisms is first done in shaker flasks (13), and then in small fermenters

(14) to determine growth conditions, and eventually in larger fermentation tanks (15). Cellular extract obtained from the fermentation process is

then separated, purified (16), and packaged (17) either for industrial use (18) or health care applications.

Health care products are first tested in animal studies (19) to demonstrate a product’s pharmacological activity and safety, in the United

States, an investigational new drug application (20) is submitted to begin human clinical trials to establish safety and efficacy. Following

clinical testing (21), a new drug application (NDA) (22) is fried with the Food and Drug Administration (FDA). When the NDA has been reviewed

and approved by the FDA the product maybe marketed in the United States (23).

SOURCE: Genentech, Inc.
Ch. 1—Summary: Issues and Options ● 7

class of molecules that control gene activity, are present in the body in only minute

quantities. Now, for the first time, they can

be recognized, isolated, characterized, and

produced in quantity.

The mere availability of a pharmacologically

active substance does not ensure its adoption in

medical practice. Even if it is shown to have

therapeutic usefulness, it may not succeed in

the marketplace.

The difficulty in predicting the economic impact is exemplified by interferon. If it is found to

be broadly effective against both viral diseases

and cancers, sales would be in the tens of billions of dollars annually. If its clinical effectiveness is found to be only against one or two

viruses, sales would be significantly lower.

At the very least, even if there are no immediate medical uses for compounds produced

by genetic engineering, their indirect impact on

medical research is assured. For the first time,

almost any biological phenomenon of medical

interest can be explored at the cellular level.

These molecules are valuable tools for understanding the anatomy and functions of cells.

The knowledge gained may lead to the development of new therapies or preventive measures

for diseases.

The chemical industry

FINDINGS

The chemical industry’s primary raw material, petroleum, is now in limited supply. Coal is

one appealing alternative; another is biomass, a

renewable resource composed of plant and animal material.

Biomass has been transformed by fermentation into organic chemicals like citric acid, ethanol, and amino acids for decades. Other organic

chemicals such as acetone, butanol, and fumaric

acid were at one time made by fermentation until chemical production methods, combined

with cheap oil and gas, proved to be more economical. In theory, most any industrial organic

chemical can be produced by a biological process.

Commercial fermentation using genetically

engineered micro-organisms offers several advantages over current chemical production

techniques.

The use of renewable resources: starches,

sugars, cellulose, and other components of

biomass can serve as the raw material for

synthesizing organic chemicals. With proper agricultural management, biomass can

assure a continuous renewable supply for

the industry.

The use of physically milder conditions:

chemical processes often require high temperatures and extreme pressures. These

conditions are energy intensive and pose a

hazard in case of accidents. Biological processes operate under milder conditions,

which are compatible with living systems.

One-step production methods: micro-organisms can carry out several steps in a synthetic process, eliminating the need for intermediate steps of separation and purification.

Decreased pollution: because biological

processes are highly specific in the reactions they catalyze, they offer control over

the products formed and decrease undesirable side-products. As a result, they produce fewer pollutants that require management and disposal,

The impact of this technology will cut across

the entire spectrum of chemical groups: plastics

and resin materials, flavors and perfumes materials, synthetic rubber, medicinal chemicals,

pesticides, and the primary products from petroleum that serve as the raw materials for the

synthesis of organic chemicals. Nevertheless,

the specific products that will be affected in

each group can only be chosen on a case-by-case

basis, with the applicability of genetics depending on a variety of factors. Crude estimates

of the expected economic impacts are in the billions of dollars per year for dozens of chemicals

within 20 years.

INDUSTRY AND MANPOWER IMPACTS

Although genetic engineering will develop

new techniques for synthesizing many substances, the direct displacement of any current

industry seems doubtful. Genetic engineering

should be considered simply another industrial

tool. Industries will probably use genetic
8 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

engineering to maintain their positions in their

respective markets. This is already illustrated

by the variety of companies in the pharmaceutical, chemical, and energy industries that have

invested in or contracted with genetic engineering firms. Some large companies are already developing inhouse genetic engineering research

capabilities.

Any predictions of the number of workers

that will be required in the production phase of

biotechnology will depend on the expected

volume of chemicals that will be produced. At

present, this figure is unknown. An estimated

$15 billion worth of chemicals maybe manufactured by biological processes. This will employ

approximately 30,000 to 75,000 workers for supervision, services, and production. Whether

this will represent a net loss or gain in the number of jobs is difficult to predict since new jobs

in biotechnology will probably displace some of

those in traditional chemical production.

Food processing industry

FINDINGS

Genetics in the food processing industry can

be used in two ways: to design micro-organisms

that transform inedible biomass into food for

human consumption or into feed for animals;

and to design organisms that aid in food processing, either by acting directly on the food

itself or by providing materials which can be

added to food.

The use of genetics to design organisms with

desired properties for food processing is an

established practice. Fermented foods and

beverages have been made by selected strains

of mutant organisms (e.g., yeasts) for centuries.

Only recently, however, have molecular technologies opened up new possibilities. In particular, large-scale availability of enzymes will

play an increasing role in food processing.

.

The applications of molecular genetics are

likely to appear in the food processing industry

in piecemeal fashion:

● Inedible biomass, human and animal

wastes, and even various industrial effluents are now being transformed into edible

●

micro-organisms high in protein content

(called single-cell protein or SCP). Its present cost of production in the United States

is relatively high, and it must compete with

cheaper sources of protein such as soybeans and fishmeal, among others.

Isolated successes can be anticipated for

the production of such food additives as

fructose (a sugar) and the synthetic sweetener aspartame, and for improvements in

SCP production.

An industrywide impact is not expected in the

near future because of several major conflicting

factors:

●

●

●

The basic knowledge of the genetic characteristics that could improve food has not

been adequately developed.

The food processing industry is conservative in its expenditures for R&D to improve

processes. Generally, only one-third to onehalf as much is allocated for this purpose as

in technologically intensive industries.

Products made by new microbial sources

must satisfy the Food and Drug Administration’s (FDA) safety regulations, which include undergoing tests to prove lack of

harmful effects. It may be possible to

reduce the amount of required testing by

transferring the desired gene into microorganisms that already meet FDA standards.

The use of genetically engineered

micro-organisms in the environment

FINDINGS

Genetically engineered micro-organisms are

being designed now to perform in three areas

(aside from agricultural uses) that require their

large-scale release into the environment:

. mineral leaching and recovery,

. enhanced oil recovery, and

● pollution control.

All of these are characterized by:

● the use of large volumes of micro-organisms,

 decreased control over the behavior and

fate of the micro-organisms,
Ch. 1—Summary: Issues and Options ● 9

the possibility of ecological disruption, and

less development in basic R&D (and more

speculation-) than in the industries in which

micro-organisms are used in a controlled

environment.

MINERAL LEACHING AND RECOVERY

Bacteria have been used to leach metals, such

as uranium and copper, from low-grade ores.

Although there is reason to believe leaching

ability is under genetic control in these organisms, practically nothing is known about the

precise mechanisms involved. Therefore, the

application of genetic technologies in this area

remains speculative. Progress has been slow in

obtaining more information, partly because

very little research has been conducted.

In addition to leaching, micro-organisms can

be used to recover valuable metals or eliminate

polluting metals from dilute solutions such as industrial waste streams. The process makes use

of the ability of micro-organisms to bind metals

to their surfaces and then concentrate them internally.

The economic competitiveness of biological

methods is still unproved, but genetic modifications have been attempted only recently. The

cost of producing the micro-organisms has been

a major consideration. If it can be reduced, the

approach might be useful.

ENHANCED OIL RECOVERY

Many methods have been tried in efforts to

remove oil from the ground when natural

expulsive forces alone are no longer effective.

Injecting chemicals into a reservoir has, in many

cases, aided recovery by changing the oil’s flow

characteristics.

Micro-organisms can produce the necessary

chemicals that help to increase flow. Theoretically, they can also be grown in the wells

themselves, producing those same chemicals in

situ. The currently favored chemical, xanthan,

is far from ideal for increasing flow. Genetic

engineering should be able to produce chemicals with more useful characteristics.

The current research approach, funded by

the Department of Energy (DOE) and independently by various oil companies, is a two-phase

process to find micro-organisms that can function in an oil reservoir environment, and then to

improve their characteristics genetically.

The genetic alteration of micro-organisms to

produce chemicals useful for enhanced oil recovery has been more successful than the alteration of micro-organisms that may be used in

situ. However, rDNA technology has not been

applied to either case. All attempts have employed artificially induced or naturally occurring mutations.

POLLUTION CONTROL

Many micro-organisms can consume various

kinds of pollutants, changing them into relatively harmless materials before they die. These

micro-organisms always have had a role in

“natural” pollution control: nevertheless, cities

have resisted adding microbes to their sewerage

systems. Although the Environmental Protection Agency (EPA) has not recommended addition of bacteria to municipal sewerage systems,

it suggests that they might be useful in smaller

installations and for specific problems in large

systems. In major marine spills, the bacteria,

yeast, and fungi already present in the water

participate in degradation. The usefulness of

added microbes has not been demonstrated.

Nevertheless, in 1978, the estimated market

of biological products for pollution control was

$2 million to $4 million per year, divided among

some 20 companies; the potential market was

estimated to be as much as $20 million per year.

To date, genetically engineered strains have

not been applied to pollution problems. Restricting factors include the problems of liability in

the event of health, economic, or environmental

damage; the contention that added organisms

are not likely to be a significant improvement;

and the assumption that selling microbes rather

than products or processes is not likely to be

profitable.

Convincing evidence that microbes could remove or degrade an intractable pollutant would

encourage their application. In the meantime,

however, these restrictions have acted to inhibit

the research necessary to produce marked improvements.
10 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

CONSTRAINTS IN USING GENETIC ENGINEERING

TECHNOLOGIES IN OPEN ENVIRONMENTS

The genetic data base for the potentially useful micro-organisms is lacking. Only the simplest methods of mutation and selection for desirable properties have been used thus far.

These are the only avenues for improvement

until more is learned about the genetic mechanisms.

Even when the scientific knowledge is available, two other obstacles to the use of genetisystems on a scale large enough to exploit their

biological activity. This will necessitate a continual dialog among microbial geneticists, geologists, chemists, and engineers; an interdisciplinary approach is required that recognizes the

needs and limitations of each discipline.

The second obstacle is ecological. Introducing

large numbers of genetically engineered microorganisms into the environment might lead to

ecological disruption or detrimental effects on

human health, and raise questions of legal liacally engineered

The first is the

micro-organisms will remain. bility.

—

need to develop engineered

Issue and Options—Biotechnology

ISSUE: How can the Federal Government promote advances in biotechnology and genetic engineering?

The United States is a leader in applying

genetic engineering and biotechnology to industry. One reason is the long-standing commitment by the Federal Government to the funding

of basic biological research; several decades of

support for some of the most esoteric basic

research has unexpectedly provided the foundation for a highly useful technology. A second

is the availability of venture capital, which has

allowed the formation of small, innovative companies that can build on the basic research.

The chief argument for Government subsidization for R&D in biotechnology and genetic

engineering is that Federal help is needed in

areas such as general (generic) research or highly speculative investigations not now being developed by industry. The argument against the

need for this support is that industry will develop everything of commercial value on its own.

A look at what industry is now attempting indicates that sufficient investment capital is

available to pursue specific manufacturing objectives. Some high-risk areas, however, that

might be of interest to society, such as pollution

control, may justify promotion by the Government, while other, such as enhanced oil recovery might might not be profitable soon enough

to attract investment by industry.

OPTIONS:

A. Congress could allocate funds specifically for

genetic engineering and biotechnology R&Din

the budget of appropriate agencies.

Congress could promote two types of programs in biotechnology: those with long-range

payoffs (basic research), and those that industry

is not willing to undertake but that might be in

the national interest.

B. Congress could establish a separate Institute

of Biotechnology as a funding agency.

The merits of a separate institute lie in the

possibility of coordinating a wide range of efforts, all related to biotechnology. On the other

hand, biotechnology and genetic engineering

cover such a broad range of disciplines that a

new funding agency would overlap the mandates of existing agencies. Furthermore, the

creation of yet another agency carries with it all

the disadvantages of increased bureaucracy and

competition for funds at the agency level.

C. Congress could establish research centers in

universities to foster interdisciplinary approaches to biotechnology. In addition, a program of grants could be offered to train scientists in biological engineering.

The successful use of biological techniques in

industry depends on a multidisciplinary approach involving biochemists, geneticists, microbiologists, process engineers, and chemists.
Ch. 1—Summary: lssues and Options ● 11

Little is now being done publicly or privately to

develop the expertise necessary.

D. Congress could use tax incentives to stimulate

biotechnology.

The tax laws could be used to stimulate biotechnology by expanding the supply of capital

for small, high-risk firms, which are generally

considered more innovative than established

firms because of their willingness to undertake

the risks of innovation. In addition to focusing

on the supply of capital, tax policy could attempt to directly increase the profitability of

potential growth companies.

A tax incentive could also be directed’ at increasing R&D expenditures. It has been suggested that companies be permitted to take tax

credits: 1) on a certain percentage of their R&D

expenses; and 2) on contributions to universities

for research.

E. Congress could improve the conditions under

which U.S. companies collaborate with academic scientists and make use of the technology developed in universities, which has been

wholly or partly supported by tax funds.

Developments in genetic engineering have

kindled interest in this option. Under legislation

that has recently passed both Houses of Congress, small businesses and universities may retain title to inventions developed under federally funded research. Currently, some Federal

agencies award contractors these exclusive

rights, while others insist on the nonexclusive

licensing of inventions.

F. Congress could mandate support for specific

research tasks such as pollution control using

microbes.

Microbes may be useful in degrading intractable wastes and pollutants. Current research,

however, is limited to isolating organisms from

natural sources or from mutated cultures. More

elaborate efforts, involving rDNA techniques or

other forms of microbial genetic exchange, will

require additional funding.

G. Most efforts could be left to industry and each

Government agency allowed to develop programs in the fields of genetic engineering and

biotechnology as it sees fit.

Generic research will probably not be undertaken by any one company. Leaving all R&D in

industry’s hands would still produce major commercial successes, but does not ensure the development of needed basic general knowledge

or the undertaking of high-risk projects.

Agriculture

The complexity of plants and animals presents a greater challenge to advances in applied

genetics than that posed by micro-organisms.

Nevertheless, the successful genetic manipulation of microbes has encouraged researchers in

the agricultural sciences. The new tools will be

used to complement, but not replace, the wellestablished practices of plant and animal

breeding.

The applications of genetics to plants

FINDINGS

It is impossible to exactly determine the extent to which applied genetics has directly contributed to increases in plant yield because of

simultaneous improvements in farm management, pest control, and cropping techniques

using herbicides, irrigation, and fertilizers.

Nevertheless, the impacts of breeding technologies have been extensive.

The plant breeder’s approach is determined

for the most part by the particular biological

factors of the crop being bred. The new genetic

technologies potentially offer additional tools to

allow development of new varieties and even

species of plants by circumventing current biological barriers to the exchange of genetic

material.

Technologies developed for classical plant

breeding and those of the new genetics should

not be viewed as being competitive; they are

both tools for effectively manipulating genetic
12 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

information. One new technology -e.g., protoplasm fusion, or the artificial fusion of two cells–

allows breeders to overcome incompatibility

between plants. But the plant that may result

still must be selected, regenerated, and evaluated under field conditions to ensure that the

genetic change is stable and that the attributes

of the new variety meet commercial requirements.

In theory, the new technologies will expand

the capability of breeders to exchange genetic

information by overcoming natural breeding

barriers. To date, however, they have not had a

widespread impact on the agricultural industry.

As a note of caution, it must be emphasized

that no plant can possess every desirable trait.

There will always have to be some tradeoff;

often quality for quantity, such as increased

protein content but decreased yield.

NEW GENETIC TECHNOLOGIES FOR

PLANT BREEDING

The new technologies fall into two categories:

those involving genetic transformations

through cell fusion and those involving the insertion or modification of genetic information

through the cloning of DNA and its vectors.

Techniques are available for manipulating

organs, tissues, cells, or protoplasts in culture;

for regenerating plants; and for testing the

genetic basis of novel traits. So far these techniques are routine only in a few species.

The approach to exploiting molecular biology

for plant breeding is similar in some respects to

the genetic manipulation of micro-organisms.

However, there is one major conceptual difA plantlet of Ioblolly pine grown in Weyerhaeuser Co.’s

tissue culture laboratory. The next step in this procedure

is to transfer the plantlet from its sterile and humid

environment to the soil

Photo credits: Weyerhaeuser Co.

A young Douglas fir tree propagated 4 years ago from a

small piece of seedling leaf tissue. Three years ago this was

at the test-tube stage seen in the Ioblolly pine photograph
Ch. 1—Summary: Issues and Options . 13

ference. In micro-organisms, the changes made

on the cellular level are the goals of the

manipulation. With crops, changes made on the

cellular level are meaningless unless they can be

reproduced in the entire plant as well. Therefore, unless single cells in culture can be

selected and grown into mature plants and the

desired traits expressed in the mature plant–

procedures which at this time have had limited

success—the benefits of genetic engineering will

not be widely felt in plant breeding.

Moderate success has been achieved for

growing cells in tissue culture into mature

plants. Tissue culture programs of commercial

significance in the United States include the

asparagus, citrus fruits, pineapples, and strawberries. Breeders have had little success, however, in regenerating mature plants of wide

agronomic importance) such as corn and wheat.

Some success can be claimed for engineering

changes to alter genetic makeup. Both the stable

integration of genetic material into a cell and

the fusion of genetically different cells are still

largely experimental techniques. Technical

breakthroughs have come on a species-byspecies basis, but key discoveries are not often

applicable to all plants. Initial results suggest

that agronomically important traits, such as

disease resistance, can be transferred from one

species to another. Limited success has also

been shown in attempts to create totally new

species by fusing cells from different genera.

Attempts to find both suitable vectors and genes

for transferring one plant genes to another are

only now beginning to show promise.

CONSTRAINTS ON USING MOLECULAR

GENETICS FOR PLANT IMPROVEMENTS

Molecular engineering has been impeded by a

lack of answers to basic questions in molecular

biology and plant physiology owing to insufficient research. Federal funding for plant molecular genetics in agriculture has come primarily

from the U.S. Department of Agriculture

(USDA) and the National Science Foundation

(NSF). In USDA, research support is channeled

primarily through the flexible competitive

grants program (fiscal year 1980 budget of $15

million) for the support of new research directions in plant biology. The total support for the

plant sciences from NSF is approximately $25

million, only $1 million of which is specifically

designated for plant genetics.

The shortage of a trained workforce is a

significant constraint. Only a few universities

have expertise in both plants and molecular biology. In addition, there are only a few people

who have the ability to work with modern molecular techniques related to whole plant problems. As a result, a business firm could easily

develop a capability in this area exceeding that

at any individual U.S. university. However, the

building of industrial laboratories and subsequent hiring from the universities could easily

deplete the expertise at the university level.

With the recent investment activity by many

bioengineering firms, this trend has already

begun; in the long-run it could have serious consequences for the quality and quantity of university research.

GENETIC VARIABILITY, CROP VULNERABILITY,

AND THE STORAGE OF GERMPLASM

Successful plant breeding is based on the

availability of genetically diverse plants for the

insertion of new genes into plants. The number

of these plants has been diminishing for a variety of reasons. However, the rate and extent of

this trend is unknown; the data simply do not

exist. Therefore, it is essential to have an adequate scientific understanding of how much genetic loss has taken place and how much germplasm (the total genetic variability available to a

species) is needed. Neither of these questions

can be answered completely at this time.

Even if-genetic needs can be adequately identified, there is disagreement about the quantity

of germplasm to collect. Furthermore, the extent to which the new genetic technologies will

affect genetic variability, vulnerability, or the

storage technologies of germplasm has not been

determined. As a result, it is currently difficult,

if not impossible, to state how much effort

should be expended by the National Germplasm

System to collect, maintain, and test new gene

resources (in this case as seed).

Finally, even if an adequate level of genetic

variability can be assessed, the real problem of

vulnerability—the practice of planting only a
14 ● Impacts Of Applied Genetics—MicrO.Organ/SrnS, Plants, and Animals

single variety—must be dealt with at an institu- nologies existed, farmers would still select only

tional or social level. Even if no genetic tech- one or a few “best” varieties for planting.

Issues and Options—Plants

ISSUE: Should an assessment be conducted to determine how much

diversity in plant germplasm

needs to be maintained?

An understanding of how much germplasm

should be protected and maintained would

make the management of genetic resources

simpler.

OPTIONS:

A. Congress could commission a study of how

much genetic variability is necessary or desirable to meet present and future needs.

A comprehensive evaluation of the National

Germplasm System’s requirements for collecting, evaluating, maintaining, and distributing

genetic resources for plant breeding and research could serve as a baseline for a further

assessment.

B. Congress could commission a study on the

need for international cooperation to manage

and preserve genetic resources both in natural

ecosystems and in repositories.

This investigation could include an evaluation

of the rate at which genetic diversity is being

lost from natural and agricultural systems along

with an estimate of the effects this loss will

have.

C. Congress could commission a study on how to

develop an early warning system to recognize

the potential vulnerability of crops.

Where high genetic uniformity still exists,

proposals could be suggested to reduce any

risks due to uniformity. Alternatively, the

avenues by which private seed companies could

be encouraged to increase the levels of genetic

diversity could be investigated.

ISSUE: What are the most appropriate

approaches in overcoming the

various technical constraint s

that limit the success of molecular genetics for plant improvement?

Although genetic information has been transferred by vectors and protoplasm fusion, DNA

transformations of commercial value have not

yet been performed. Molecular engineering has

been impeded by the lack of vectors that can

transfer novel genetic material into plants,

by insufficient knowledge about which genes

would be useful for breeding purposes, and by

a lack of understanding of the incompatibility of

chromosomes from diverse sources. Another

impediment has been the lack of researchers

from a variety of disciplines.

OPTIONS:

A.

B.

c.

The level of funding could be increased for

plant molecular genetics research supported

by NSF and USDA.

Research units devoted to plant molecular genetics could be established under the auspices

of the National Institutes of Health (NIH), with

emphasis on potential pharmaceuticals derived from plants.

An institute for plant molecular genetics could

be established under the Science and Education Administration at USDA that would include multidisciplinary teams to consider both

basic research questions and direct applications of the technology to commercial needs

and practices.

The discoveries of molecular plant genetics

will be used in conjunction with traditional

breeding programs. Hence, each of the three

options could require additional appropriations

for agricultural research.
Ch. 1—Summary: Issues adoptions ● 1 5

Advances in reproductive biology and

their effects on animal improvement

FINDINGS

Much improvement can be made in the germplasm of all major farm animal species using existing technology. The expanded use of artificial

insemination (AI) with stored frozen sperm, especially in beef cattle, would benefit both producers and consumers. New techniques for synchronizing estrus should encourage the wider

use of AI. Various manipulations of embryos

will find limited use in producing breeding

stocks, and sex selection and twinning techniques should be available for limited applications within the next 10 to 20 years.

The most important technology in reproductive physiology will continue to be AI. Due in

part to genetic improvement, the average milk

yield of cows in the United States has more than

doubled in the past 30 years, while the total

number of milk cows has been reduced by more

than half. AI along with improved management

and the availability and use of accurate progeny

records on breeding stock have caused this

great increase. (See figure 3.)

The improvement lags behind what is theoretically possible. In practice, the observed increase is about 100 lb of milk per cow per year,

while a hypothetical breeding program using AI

would result in a yearly gain of 220 lb of milk

per cow. The biological limits to this rate of gain

are not known.

In comparison with dairy cattle, the beef cattle industry has not applied AI technology widely. Only 3 to 5 percent of U.S. beef is artificially

inseminated, compared to 60 percent of the

dairy herd. This low rate for beef cattle can be

explained by several factors, including management techniques (range v. confined housing)

and the conflicting objectives of individual

breeders, ranchers, breed associations, and

commercial farmers.

The national calf crop—calves alive at weaning as a fraction of the total number of cows exposed to breeding each year—is only 65 to 81

percent. An improvement of only a few percentage points through AI would result in savings of

hundreds of millions of dollars to producers and

consumers.

Coupled with a technology for estrus-cycle

regulation, the use of AI could be expanded for

both dairy and beef breeding. Embryo transfer

technology, already well-developed but still

costly, can be used to produce valuable breeding stock. Sexing technology, which is not yet

perfected, would be of enormous benefit to the

beef industry because bulls grow faster than

heifers.

In the case of animals other than cows:

●

●

●

●

●

Expanded use of AI for swine production

will be encouraged by the strong trend to

confinement housing, although the poor

ability of boar sperm to withstand freezing

will continue to be a handicap.

The benefits of applied genetics have not

been realized in sheep production because

neither AI nor performance testing has

been used. As long as the use of AI continues to be limited by the inability to

freeze semen and by a lack of agents on the

market for synchronizing estrus, no rapid

major gains can be expected.

Increasing interest in goats in the United

States and the demand for goat products

throughout the world, should encourage

attention to the genetic gains that the use

of AI and other technologies make possible.

Poultry breeders will continue to concentrate on improved egg production, growth

rate, feed efficiency, and reduced body fat

and diseases. The use of frozen semen

should increase as will the use of AI and

dwarf broiler breeders.

Genetics applied to production of fish,

mollusks, and crustaceans in either natural

environments or manmade culture systems

is only at the rudimentary stage.

Breeders must have reliable information

about the genetic value of the germplasm they

are considering introducing. Since farmers do

not have the resources to collect and process

data on the performance of animals other than

those in their own herds, they must turn to outside sources. The National Cooperative Dairy

Herd Improvement Program (NCDHIP) is a mod-
16 . The Impacts of Genetics:Applicatjions to Micro-Organisms, Plants, and Animals

Figure 3.-The Way the Reproductive Technologies Interrelate

Bull

Sperm

Sexed?

Frozen?

Recipient Herd: Synchronized estrus

Embryo transfer

Each get two for twinning

Artificial

insemination

Superovulated

cow

Recovered

embryos

Sexed?

Calves

Photo Credit: Science

These 10 calves from Colorado State University were the

result of superovulation, in vitro culture, and transfer to

the surrogate mother cows on the left. The genetic

mother of all 10 calves is at upper right

SOURCE: Office of Technology Assessment.
Ch. 1—Summary: Issues and Options . 17

el information system and could be adapted to

other species.

Selection—deciding which animals to mate

—is the breeder’s most basic tool. When going

outside his herd to purchase new germplasm,

the breeder needs impartial information about

the quality of the available germplasm. NCDHIP

had recorded 2.8 million of the 10.8 million U.S.

dairy cattle in 1979. In 1978, cows enrolled in

the official plans of NCDHIP outproduced cows

not enrolled by 5,000 lb of milk per cow, representing 52 percent more milk per lactation.

No comparable information system exists for

other types of livestock. Beef bulls, for example,

continue to be sold to a large extent on the basis

of pedigrees, but with relatively little objective

information on their genetic merit. Data on

dairy goats in the United States became available through NCDHIP for the first time in late

1980. No nationwide information systems exist

for other species, although pork production in

the United States would greatly benefit from a

national swine testing program.

The more esoteric methods of genetic manipulation will probably have little impact on the

production of animals or animal products within the next 10 years. other in vitro manipulations, such as cloning, cell fusion, the production of chimeras, and the use of rDNA techniques, will continue to be of intense interest,

especially for research purposes. It is less likely,

however, that they will have widespread practical effects on farm production in this century.

Each technique requires more research and

refinement. Until specific genes of farm animals

can be identified and located, no direct gene

manipulation will be practicable. In addition

this will be difficult because most traits of importance are due to multiple genes.

Issue and Options—Animals

ISSUE: How can the Federal Government improve the germplasm of

major farm animal species?

OPTIONS:

A. Programs like the NCDHIP could have increased governmental participation and funding. The efforts of the Beef Cattle Improvement Federation to standardize procedures

could receive active support, and a similar information system for swine could be established.

The fastest and least expensive way to upgrade breeding stock in the United States is

through effective use of information. Computer

technology, along with a network of local repre-

* *

The wide variety of applications for genetic

engineering is summarized in figure 4. Genetics

can be used to improve or increase the quality

and output of plants and animals for direct use

by man. Alternatively, materials can be extracted from plants and animals for use in food,

chemical, and pharmaceutical industries.

sentatives for data collecting, can provide the

individual farmer or breeder with accurate information on the available germplasm so that he

can make his own breeding decisions.

This option implies that the Federal Government would play such a role in new programs,

and expand its role in existing ones.

B. Federal funding could be increased for basic

research in total animal improvement.

This option, in contrast to option A, assumes

that it is necessary to maintain or expand basic

R&D to generate new knowledge that can be

applied to the production of improved animals

and animal products.

*

Biological materials can also be converted to

useful products. In this process, genetic engineering can be used to develop micro-organisms

that will carry out the conversions. Therefore,

genetic manipulation cannot only provide more

or better biological raw materials but can also

aid in their conversion to useful products.
18 . The Impacts of Genetics: Applications to Micro-Organisms, Plants, and Animals

Figure 4.—AppIications of Genetics

I

AGRICULTURAL

INDUSTRY I

Plants

Animals

(lncrease/Improve, Output)

SOURCE: Office of Technology Assessment.

Institutions and society

Regulation of genetic engineering

FINDINGS

No evidence exists that any unexpectedly

harmful genetically engineered organism has

been created. Yet few experts believe that molecular genetic techniques are totally without

risk to health and the environment. Information

that has proved useful in assessing the risks

from these techniques has come from three

sources: experiments designed specifically to

test the consequences of working with rDNA,

experiments designed for other purposes but

relevant to rDNA, and scientific meetings and

workshops.

A program of risk assessment was established

at NIH in 1979 to conduct experiments and collate relevant information. It assesses one form

of genetic engineering, rDNA. On the basis of

these data, conjectured, inadvertent risk is

generally regarded as less likely today than

originally suspected. Risk due to the manipulation of genes from organisms known to be hazardous is considered to be more realistic. Therefore, microbiological safety precautions that are
Ch. 1—Summary: lssues and Options . 19

appropriate to the use of the micro-organisms

serving as the source of DNA are required. Nevertheless, it has not been demonstrated that

combining those genes in the form of rDNA is

any more hazardous than the original source of

the DNA.

Perceptions of the nature, magnitude, and acceptability of the risk differ. In addition, public

concern has been expressed about possible

long-range impacts of genetic engineering. In

this context, the problem facing the policymaker is how to address the risk in a way that

accommodates the perceptions and values of

those who bear it.

The NIH Guidelines for Research Involving

Recombinant DNA Molecules and existing Federal laws appear adequate in most cases to deal

with the risks to health and the environment

presented by genetic engineering. However, the

Guidelines are not legally binding on industry,

and no single statute or combination will clearly

cover all foreseeable commercial applications of

genetic engineering.

The Guidelines are a flexible evolving oversight mechanism that combines technical expertise with public participation. They cover the

most widely used and possibly risky molecular

genetic technique–rDNA–prohibiting experiments using dangerous toxins or pathogens and

setting containment standards for other potentially hazardous experiments. Although compliance is mandatory only for those receiving NIH

funds, other Federal agencies follow them, and

industry has proclaimed voluntary compliance.

Rare cases of noncompliance have occurred in

universities but have not posed risks to health

or the environment. As scientists have learned

more about rDNA and molecular genetics, the

restrictions have been progressively and substantially relaxed to the point where 85 percent

of the experiments can now be done at the

lowest containment levels, and virtually all

monitoring for compliance now rests with approximately 200 local self-regulatory committees called institutional biosafety committees

(IRCs). (See table 1.)

Under the Guidelines, NIH serves an important oversight role by sponsoring risk assessTable I.—Containment Recommended by the

National institutes of Health

Biological—Any combination of vector and host must be

chosen to minimize both the survival of the system

outside of the laboratory and the transmission of the

vector to nonlaboratory hosts. There are three levels

of biological containment:

HVl– Requires the use of Escherichia coli K12 or

other weakened strains of micro-organisms that

are less able to live outside the laboratory.

HV2– Requires the use of specially engineered strains

that are especially sensitive to ultraviolet light,

detergents, and the absence of certain

uncommon chemical compounds.

HV3– No organism has yet been developed that can

qualify as HV3.

Physical—Special laboratories (P1-P4)

Pl— Good laboratory procedures, trained personnel,

wastes decontaminated

P2– Biohazards sign, no public access, autoclave in

building, hand-washing facility

P3– Negative pressure, filters in vacuum line, class II

safety cabinets

P4– Monolithic construction, air locks, all air

decontaminated, autoclave in room, all

experiments in class Ill safety cabinets (glove

box), shower room

SOURCE: Office of Technology Assessment.

ment programs, certifying new host-vector systems, serving as an information clearinghouse,

and coordinating Federal and local activities.

Limitations in NIH’s oversight are that: it lacks

legal authority over industry; its procedures for

advising industry on large-scale projects have

not incorporated sufficient expertise on largescale fermentation technology; its monitoring

for either compliance or consistent application

of the Guidelines by individuals or institutions is

virtually nonexistent; and it has not systematically evaluated other techniques, such as cell fusion, that might present risks.

Federal laws on health and environment will

cover most commercial applications of genetic

engineering. Products such as drugs, chemicals,

and foods can be regulated by existing laws.

However, uncertainty exists about the regulation of either production methods using engineered micro-organisms or their intentional

release into the environment, when the risk has

not been clearly demonstrated. While a broad

interpretation of certain statutes, such as the

Occupational Safety and Health Act and the

Toxic Substances Control Act, might cover these
20 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

situations, regulatory actions based on such in- substantial regulatory authority over commerterpretations could be challenged in court. In cial genetic engineering have not yet officially

any evident those agencies that could have acted to assert that authority.

Issue and Options—Regulation

ISSUE: How could Congress address the

risks presented by genetic engineering?

OPTIONS:

A. Congress could maintain the status quo by letting NIH and the regulatory agencies set the

Federal policy.

Congress might determine that legislation to

remedy the limitations in current Federal oversight would result in unnecessary and burdensome regulation. No known harm to health or

the environment has occurred under current

regulation. Also the agencies generally have the

legal authority and expertise to adapt to most

new problems posed by genetic engineering.

The disadvantages are the lack of a centralized, uniform Federal response to the problem,

and the possibility that risks associated with

commercial applications will not be adequately

addressed. Conflicting or redundant regulations

of different agencies would result in unnecessary burdens on those regulated.

B. Congress could require that the Federal Interagency Advisory Committee on Recombinant

DNA Research prepare a comprehensive report on its members’ collective authority to

regulate rDNA and on their regulatory intentions.

The Industrial Practices Subcommittee of this

Committee has been studying agency authority

over commercial rDNA activities. Presently,

there is little official guidance on regulatory requirements for companies that may soon market products made by rDNA methods. A congressionally mandated report would ensure full

consideration of these issues by the agencies

and expedite the process. On the other hand,

the agencies are studying the situation, which

must be done before they can act. Also, it is

often easier and more efficient to act on each

case as it arises, rather than on a hypothetical

basis before the fact.

C. Congress could require that all recombinant

DNA activity be monitored for a limited number of years.

This represents a “wait and see” position by

Congress and the middle ground between the

status quo and full regulation. It recognizes and

balances the following factors: 1) the absence of

demonstrated harm to human health or the environment from genetic engineering; 2) the continuing concern that genetic engineering presents risks; 3) the lack of sufficient knowledge

and experience from which to make a final judgment; 4) the existence of an oversight mechanism that seems to be working well, but that

has clear limitations with respect to commercial

activities; 5) the virtual abolition of Federal

monitoring of rDNA activities by recent amendments to the Guidelines; and 6) the expected increase in commercial genetic engineering.

This option would provide a data base that

could be used for: 1) determining the effectiveness of voluntary compliance with the

Guidelines by industry, and mandatory compliance by Federal grantees; 2) determining the

quality and consistency of the local self-regulatory actions; 3) continuing a formal risk assessment program; 4) identifying vague or conflicting provisions of the Guidelines for revision; 5)

identifying emerging trends or problems; and 6)

tracing any long-term adverse impacts on health

or the environment to their sources.

The obvious disadvantage of this option

would be the required paperwork and effort by

scientists, universities, corporations, and the

Federal Government.

D. Congress could make the NIH Guidelines applicable to all rDNA work done in the United

States.

This option would eliminate any concern

about the effectiveness of voluntary compliance

with the Guidelines, and it has the advantage of
Ch. 1—Summary: Issues adoptions ● 2 1

using an existing oversight mechanism. The major changes that would have to be made in the

area of enforcement. Present penalties for noncompliance—suspension or termination of research funds-are obviously inapplicable to industry. In addition, procedures for monitoring

compliance would have to be strengthened.

The main disadvantage of this option is that

NIH is not a regulatory agency. Since NIH has

traditionally viewed its mission as promoting

biomedical research, it would have a conflict of

interest between regulation and promotion.

One of the regulatory agencies could be given

the authority to enforce the Guidelines.

E. Congress could require an environmental impact statement and agency approval before

any genetically engineered organism is intentionally released into the environment.

There have been numerous cases where an

animal or plant species has been introduced into

a new environment and has spread in an uncontrolled and undesirable fashion. Yet in pollution

control, mineral leaching, and enhanced oil

recovery, it might be desirable to release large

numbers of engineered micro-organisms into

the environment.

The Guidelines currently prohibit deliberate

release of any organism containing rDNA without approval of NIH. One disadvantage of this

prohibition is that it lacks the force of law.

Another is that approval may be granted on a

finding that the release would present “no significant risk to health or the environment;” a

tougher or more specific standard may be desirable.

A required study of the possible consequences of releasing a genetically engineered

organism would be an important step in ensuring safety. An impact statement could be filed

before permission is granted to release the

organism. However, companies and individuals

might be discouraged from developing useful

organisms if this process became too burdensome and costly.

F. Congress could pass legislation regulating all

types and phases of genetic engineering from

research through commercial production.

This option would deal comprehensively and

directly with the risks of novel molecular

genetic techniques. A specific statute would

eliminate the uncertainties over the extent to

which present law covers particular applications of genetic engineering and any concerns

about the effectiveness of voluntary compliance

with the Guidelines. Alternatively, the legislation could take the form of amending existing

laws to clarify their applicability to genetic

engineering.

Other molecular genetic techniques, while

not as widely used and effective as rDNA, raise

similar concerns. Of the current techniques, cell

fusion is the prime candidate for being treated

like rDNA in any regulatory framework. No risk

assessment of this technique has been done, and

no Federal oversight exists.

The principal argument against this option is

that the current system appears to be working

fairly well, and the limited risks of the techniques may not warrant the significantly increased regulatory burden that would result

from such legislation.

G. Congress could require NIH to rescind the

Guidelines.

Deregulation would have the advantage of allowing money and personnel currently involved

in implementing the Guidelines at the Federal

and local levels to be used for other purposes.

There are several reasons for retaining the

Guidelines. Sufficient scientific concern exists

for the Guidelines to prohibit certain experiments and to require containment for others.

Most experiments can be done at the lowest,

least burdensome containment levels. NIH is

serving an important role as a centralized oversight and information coordinating body, and

the system has been flexible enough in the past

to liberalize the restrictions as evidence indicated lower risk than originally thought.

H. Congress could consider the need for regulating work with all hazardous micro-organisms

and viruses, whether or not they are genetically engineered.

It was not within the scope of this study to examine this issue, but it is an emerging one that

Congress may wish to consider.

76-565 0 - 81 - 3
22 . Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

Patenting living organisms

On June 16, 1980, in a 5-to-4 decision, the Supreme Court ruled that a human-made microorganism was patentable under Federal patent

statutes. The decision while hailed by some as

assuring this country’s technological future was

at the same time denounced by others as creating Aldous Huxley’s Brave New World. It will do

neither.

FINDINGS

1. Meaning and Scope of the Decision.—The

decision held that a patent could not be denied

on a genetically engineered micro-organism that

otherwise met the legal requirements for patentability solely because it was alive. It was

based on the Court’s interpretation of a provision of the patent law which states that a patent

may be granted on “. . . any new and useful . . .

manufacture, or composition of matter. . . .“ (35

U.S.c. $101)

It is uncertain whether the case will serve as

a legal precedent for patenting more complex

organisms. Such organisms, however, will probably not meet other legal prerequisites to patentability that were not at issue here. In any event,

fears that the case would be legal precedent

sometime in the distant future for patenting human beings are unfounded because the 13th

amendment to the Constitution absolutely prohibits ownership of humans.

2. Impact on the Biotechnology Industry.—The

decision is not crucial to the development of the

industry. It will stimulate innovation by encouraging the dissemination of technical information that otherwise would have been maintained as trade secrets because patents are public documents that fully describe the inventions.

In addition, the ability to patent genetically engineered micro-organisms will reduce the risks

and uncertainties facing individual companies

in the commercial development of those organisms and their products, but only to a limited

degree because reasonably effective alternatives exist. These are: 1) maintaining the organisms as trade secrets; 2) patenting microbiological processes and their products; and 3) patenting inanimate components of micro-organisms, such as genetically engineered plasmids.

3. Impact on the Patent Law and the Patent and

Trademark Office.— Because of the complexity,

reproducibility, and mutability of living organisms, the decision may cause some problems

for a body of law designed more for inanimate

objects than for living organisms. It raises questions about the proper interpretation and application of the patent law requirements of novelty, nonobviousness, and enablement. In addition, it raises questions about how broad the

scope of patent coverage on important microorganisms should be, and about the continuing

need for two statutes, the Plant Patent Act of

1930 and the Plant Variety Protection Act of

1970. These uncertainties could result in increased litigation, making it more difficult and

costly for owners of patents on living organisms

to enforce their rights.

The impact on the Patent and Trademark Office is not expected to be significant in the next

few years. Although the number of patent applications on micro-organisms have almost

doubled during 1980, the approximately 200

pending applications represent less than 0.2

percent of those processed each year by the Office. While the number of such applications is

expected to increase in the next few years

because of of the decision and developments in

the field, the Office should be able to accommodate the increase. A few additional examiners may be needed. ,

4. Impact on Academic Research.—Because the

decision may encourage academic scientists to

commercialize the results of their research, it

may inhibit the free exchange of information,

but only if scientists rely on trade secrecy

rather than patents to protect their inventions

from competitors in the marketplace. In this respect, it is not clear how molecular biology differs from other research fields with commercial

potential.
Ch. 1—Summary: Issues and Options ● 23

Issue and Options—Patenting Living Organisms

ISSUE: To what extent could Congress

provide for or prohibit the patent ing of living organisms?

OPTIONS:

The Supreme Court stated that it was undertaking only the narrow task of determining

whether or not Congress, in enacting the patent

statutes, had intended a manmade micro-organism to be excluded from patentability solely

because it was alive. Moreover, the opinion

specifically invited Congress to overrule the

decision if it disagreed with the Court’s interpretation. Congress can act to resolve the questions left unanswered by the Court, overrule

the decision, or develop a comprehensive statutory approach. Most importantly, Congress can

draw lines; it can decide which organisms, if

any, should be patentable.

A. Congress could maintain the status quo.

Congress could choose not to address the

issue of patentability and allow the law to be

developed by the courts. The advantage of this

option is that issues will be addressed as they

arise, in the context of a tangible, nonhypothetical case.

There are two disadvantages to this option: a

uniform body of law may take time to develop;

and the Federal judiciary is not designed to take

sufficient account of the broader political and

social interests involved.

B. Congress could pass legislation dealing with

the specific legal issues raised by the Court's

decision.

Many of the legal questions are so broad and

varied that they do not readily lend themselves

to statutory resolution, The precise meaning of

the requirements for novelty, nonobviousness,

and enablement as applied to biological inventions will be most readily developed on a caseby-case basis by the Patent Office and the

Federal courts. On the other hand, some questions are fairly narrow and well-defined; thus,

they could be better resolved by statute. The

most important question is whether there is a

continuing need for the two plant protection

acts that grant ownership rights to plant

breeders who develop new and distinct

varieties of plants.

C. Congress could mandate a study of the Plant

Patent Act of 1930 and the Plant Variety Protection Act of 1970.

These Acts could serve as models for studying

the broader, long-term potential impacts of

patenting living organisms. Such a study would

be timely not only because of the Court’s decision, but also because of allegations that the

Acts have encouraged the planting of uniform

varieties, loss of genetic diversity, and increased

concentration in the plant breeding industry.

D. Congress could prohibit patents either on any

living organism or on organisms other than

those already subject to the plant protection

Acts.

By prohibiting patents on any living organisms, Congress would be accepting the

arguments of those who consider ownership

rights in living organisms to be immoral, or who

are concerned about other potentially adverse

impacts of such patents, A total prohibition

would slow but not stop the development of

molecular genetic techniques and the biotechnology industry because there are severa] good

alternatives for maintaining exclusive control of

biological inventions. Development would be

slowed primarily because information that

might otherwise become public would be

withheld as trade secrets. A major consequence

would be that desirable products would take

longer to reach the market.

Alternatively, Congress could overrule the

Supreme Court’s decision by amending the patent law to prohibit patents on organisms other

than the plants covered by the two statutes

mentioned in option C. This would demonstrate

congressional intent that living organisms could

be patented only by specific statute.

E. Congress could pass a comprehensive law covering any or all organisms (except humans).

This option recognizes that Congress can

draw lines where it sees fit in this area. It could

specifically limit patenting to micro-organisms,
24 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

or it could encourage the breeding of agriculturally important animals by granting patent

rights to breeders of new and distinct breeds. In

the interest of comprehensiveness and uniformity, one statute could cover plants and all other

organisms that Congress desires to be patentable.

Genetics and society

FINDINGS

Continued advances in science and technology are beginning to provide choices that strain

human value systems in areas where previously

no choice was possible. Existing ethical and

moral systems do not provide clear guidelines

and directions for those choices. New programs,

both in public institutions and in the popular

media, have been established to explore the

relationships among science, technology, society, and value systems, but more work needs to

be done.

Genetics-and other areas of the biological

sciences—have in common a much closer relationship to certain ethical questions than do

most advances in the physical sciences or

engineering. The increasing control over the

characteristics of organisms and the potential

for altering inheritance in a directed fashion

raise again questions about the relationship of

humans to each other and to other living things.

People respond in different ways to this potential; some see it (like many predecessor developments in science) as a challenging opportunity,

others as a threat, and still others respond with

vague unease. Although many people cannot articulate fully the basis for their concern, ethical,

moral, and religious reasons are often cited.

The public’s increasing concern about the advance of science and impacts of technology has

led to demands for greater participation in decisions concerned with scientific and technological issues, not only in the United States but

throughout the world. The demands imply new

challenges to systems of representative government. In every Western country, new mechanisms have been devised for increasing citizen

participation.

The public has already become involved in

decisionmaking with regard to genetics. As the

science develops, additional issues in which the

public will demand involvement can be anticipated for the years ahead. The question then becomes one of how best to involve the public in

decisionmaking.

Issues and Options—Genetics and Society

Issue: How should the public be involved in determining policy related to new applications of genetics?

Because public demands for involvement are

unlikely to diminish, ways to accommodate

these demands must be considered.

OPTIONS:

A.

B.

Congress could specify that public opinion

must be sought in formulating all major policies concerning new applications of genetics,

including decisions on the funding of specific

research projects. A “Public Participation

Statement” could be mandated for all such

decisions.

Congress could maintain the status quo, allowing the public to participate only when it

decides to do so on its own initiative.

If option A were followed, there would be no

cause for claiming that public involvement was

inadequate (as occurred after the first set of

Guidelines for Recombinant DNA Research was

promulgated). Option A poses certain problems:

How to identify a major policy and at what stage

public involvement would be required. Should

it take place only when technological development and application are imminent, or at the

basic research stage?

Option B would be less cumbersome to effect.

It would permit the establishment of ad hoc

mechanisms when necessary.
—

Ch. 1—Summary: Issues adoptions . 25

ISSUE:

There

too little

How can the level of publi c

knowledge concerning genetics

and its potential be raised?

are some educators who believe that

time is spent on genetics within the

traditional educational system. outside the

traditional school system, a number of sources

may contribute to increased public understanding of science and the relationship between

science and society.

Efforts to increase public understanding

should, of course, be combined with carefully

designed evaluation programs so that the effectiveness of a program can be assessed.

OPTIONS:

A. Programs could be developed to increas e

public understanding of science and the relationship between science, technology, and

society.

Public understanding of science in today's

world is essential, and there is concern about

the adequacy of the public's knwledge.

B. programs could be established to monitor the

level of public understanding of genetics and

of science in general, and to determine whether public concern with decisionmaking in

science and technology is increasing.

Selecting this option would indicate that

there is need for additional information, and

that Congress is interested in involving the

public indeveloping science policy.

C. The copyright laws could be amended to permit schools to videotape television programs

for educational purposes.

Under current copyright law, videotaping television programs as they are being broadcast

may infringe on the rights of the program’s

owner, generally its producer. The legal status

of such tapes is presently the subject of litigation.

In favor of this option, it should be noted that

many of the programs are made at least in part

with public funds. Removing the copyright constraint on schools would make these programs

more available for another public good, education. On the other hand, this option could have

significant economic consequences to the copyright owner, whose market is often limited to

educational institutions.

ISSUE: Should Congress begin preparing now to resolve issues that

have not yet aroused much public debate but which may in the

future?

As scientific understanding of genetics and

the ability to manipulate inherited characteristics develops, society may face some difficult

questions that could involve tradeoffs between

individual freedom and the needs of society.

This will be increasingly the case as genetic

technologies are applied to humans. Developments are occurring rapidly. Recombinant DNA

technology was developed in the 1970’s. In the

spring of 1980, investigators succeeded in the

first gene replacement in mammals; in the fall

of 1980, the first gene substitution in humans

was attempted.

Although this study was restricted to nonhuman applications, many people assume from

these and other examples that what can be done

with lower animals can be done with humans

and will be. Therefore, some action might be

taken to better prepare society for decisions on

the application of genetic technologies to

humans.

OPTIONS:

A.

B.

A commission could be established to identify

central issues, the probable time frame for application of various genetic technologies to

humans, and the probable effects on society,

and to suggest courses of action. The commission might also consider the related area of

how participatory democracy might be combined with representative democracy in decisionmaking.

The life of the President Commission could

be extended for the study of Ethical problems

in Medicine and Biomedical and Behavioral Research, for the purpose of addressing these

issues.
26 . Impacts of Applied Genetics—Micro-Organisms, plants, and Animals

This 1 l-member ‘Commission was established

in November 1978 and terminates on December

31, 1982. It could be asked to broaden its coverage to additional areas. This would require that

the life span of the commission be extended and

additional funds be appropriated.

A potential disadvantage to using the existing

commission to address societal issues associated

with genetic engineering is that a number of

issues already exist, and more are likely to arise

in the years ahead. Yet there are also other

issues in medicine and biomedical and behavioral research not associated with genetic engineering that also need review. Whether all

these issues can be addressed by one commission should be considered. Comments from the

existing commission would assist in deciding the

most appropriate course of action.
Chapter 2

Introduction
Chapter 2

Page

The Origins of Genetics. . . . . . . . . . . . . . . . . , . . 29

(Genetics in the 20th Century. . . . . . . . . . . . . . . . 33

The Riddle of the Gene , . . . . . . . , . . . . . , . . . . . 33

The Genetic Code . . . . . . . . . . . . . . . . . . . . . . . , .37

Developing Genetic “technologies . . . . . . . , . . . . 39

The Basic Issues . . . . . . . . . . . . . . . . . . . . . . . . . 43

How Will Applied Genetics Be Used? . . . . . . . . 43

What Are the Dangers? . . . . . . . . . . . . . . . , . . 43

Figures

Figure No. Page

5. The Inheritance Pattern of Pea Color. . . . . , , 30

6. Chromosomes . . . . . , . . . . . . . . . . . . . . . . . . 32

7. The Griffith Experiment . . . . . . . . . . . . . . . . 34

8. The Structure of DNA . . . . . . . . . . . . . . . . . 36

9. Replication of DNA . . . . . . . . . . . . . . , . . . . 37

10. The Genetic Code . . . . . . . . . . . . . . . . . . . , 38

11. The Expression of Genetic Information in

the Cell . . . . . . . . . . . . . . . . . . . . . . . . . . , 39

12. Transduction: The Transfer of Genetic

Material in Bacteria by Means of Viruses . . . . 39

13. Conjugation: The Transfer of Genetic

Material in Bacteria by Mating. . . . . . . . . . . . 40

14. Recombinant DNA: The Technique of

Recombining Genre From One Species With

Those From Another , ! . . . . . . . , . . . . . . . . . 41

15. An Example of How the Recombinant DNA

Technique May Be Used To Insert New

Genes Into Bacterial Cells. . . . . . . . . , . . . . . 42
Chapter 2

Introduction

Humankind is gaining an increasing understanding of heredity and variation among living

things—the science of genetics. This report examines both the critical issues arising from the

science and technologies that spring from genetics, and the potential impacts of these advances on society. They are the most rapidly

progressing areas of human knowledge in the

world today.

Genetic technologies exist only within the

larger context of a maturing science. The key to

planning for their potential is understanding

not simply a particular technology, or breeding

program, or new opportunity for investment,

but how the field of genetics works and how it

interacts with society as a whole.

The technologies that this report assesses can

be expected to have pervasive effects on life in

the future. They touch on the most fundamental and intimate needs of mankind: health care,

supplies of food and energy, and reproduction.

At the same time, they trigger concerns in areas

The origins of genetics

For the past 10,000 years, a period encompassing less than one-half of 1 percent of man’s

time on Earth, the human race has developed

under the impetus of applied genetics. As techniques for planning, cultivating, and storing

crops replaced subsistence hunting and foraging, the character of humanity changed as well.

From the domestication of animals to the development of permanent settlements, from the rise

of modern science to the dawn of biotechnology, the genetic changes that mankind has

directed have, in turn, affected the nature of his

society.

Applied genetics depends on a fundamental

principle–that organisms both resemble and

differ from their parents. It must have required

great faith on the part of Neolithic man to bury

equally as important: the dwindling supplies of

natural resources, the risks involved in basic

and applied scientific research and development, and the nature of innovation itself.

As always, some decisions concerning the use

of the new technologies will be made by the

marketplace, while others will be made by various institutions, both public and private. In the

coming years, the public and its representatives

in Congress and other governmental bodies will

be called on to make difficult decisions because

of society’s knowledge about genetics and its

capabilities.

This report does not make recommendations

nor does it attempt to resolve conflicts. Rather,

it clarifies the bases for making judgments by

defining the likely impacts of a group of technologies and tracing their economic, societal, legal,

and ethical implications. The new genetics will

be influential for a long time to come. Although

it will continue to change, it is not too early to

begin to monitor its course.

perfectly good grain during one season in the

hope of growing a new crop several months

later–faith not only that the seed would indeed

return, but that it would do so in the form of the

same grain-producing crop from which it had

sprung. This permanence of form from one

generation to the next has been scientifically

understood only within the past century, but

the understanding has transformed vague beliefs in the inheritance of traits into the science

of genetics, and rule-of-thumb animal and plant

breeding into the modern manipulations of

genetic engineering.

The major conceptual boost for the science

of genetics required a shift in perspective,

from the simple observation that characteristics

29
30 . lmpacts of Applied Genetics—Micro-Organisms, Plants, and Animals

passed from parents to offspring, to a study of

the underlying agent by which this transmission

is accomplished. That shift began in the garden

of Gregor Mendel, an obscure monk in mid-19th

century Austria. By analyzing generations of

controlled crosses between sweet pea plants,

Mendel was able to identify the rudimentary

characteristics of what was later termed the

gene.

Mendel reasoned that genes were the vehicle

and repository of the hereditary mechanism,

and that each inherited trait or function of an

organism had a specific gene directing its development and appearance. An organism’s observable characteristics, functions, and measurable

properties taken together had to be based somehow on the total assemblage of its genes.

Mendel’s analysis showed that the genes of

his pea plants remained constant from one generation to the next, but more importantly, he

found that genes and observable traits were not

simply matched one-for-one. There were, in

fact, two genes involved in each trait, with a

single gene contributed by each parent. When

the genes controlling a particular trait are identical, the organism is homozygous for that trait;

if they are not, it is heterozygous.

In the Mendelian crosses, homozygous plants

always retained the expected characteristics.

But heterozygous plants did not simply display a

mixture of their different genes; one of the two

tended to predominate. Thus, when homozy -

gous yellow-seed peas were crossed with homozygous green-seed plants, all the offspring were

now heterozygous for seed color, possessing a

“green” gene from one parent and a “yellow”

from the other. Yet all of them turned out to be

indistinguishable from the yellow-seed parent:

Yellow-seed color in peas was dominant to

green.

But even though the offspring resembled

their dominant parent, they could be shown to

contain a genetic difference. For when the heterozygotes were now crossed with each other, a

certain number of recessive green-seed plant

again appeared among the offspring. This occurred whenever an offspring was endowed

with a pair of genes that was homozygous for

the green-seed trait—and it occurred at a rate

consistent with the random selection of one of

two genes from each parent for passage to the

new generation. (See figure 5.)

Genes were real—Mendel’s work made that

clear. But where were they located, and what

were they? The answer, lay within the nucleus

of the cell. Unfortunately, most of the contents

of the nucleus were unobtainable by biologists

in Mendel’s time, so his published findings were

ignored. Only during the last decades of the

19th century did improved microscopes and

new dyes permit cells to be observed with an

acuity never before possible. And only by the

Figure 5.-The Inheritance Pattern of Pea Color

Y = yellow gene g = green gene

Each parent contributes only one seed-color gene to the offspring. When the two YY and gg homozygotes are crossed,

the genetic composition of all offspring is Yg:

Offspring

All Yg offspring are heterozygous, and all have yellow

seeds, indicating that the Y yellow gene is dominant over

the g green gene.

When these Yg heterozygotes are crossed with each other:

Thus, 3/4 of these offspring will have yellow seeds, but their

individual genetic composition, YY of Yg, maybe different.

SOURCE: Office of Technology Assessment.
Ch. 2—Introduction ● 37

beginning of the 20th century did scientists

rediscover Mendel’s work and begin to appreciate fully the significance of the cell nucleus

and its contents.

Even in the earliest microscopic studies,

however, certain cellular components stood

out; they were deeply stained by added dye. As

a result, they were dubbed ‘(colored bodies, ” or

chromosomes. Chromosomes were seen relatively rarely in cells, with most cells showing

just a central dark nucleus surrounded by an

extensive light grainy cytoplasm. But periodically the nucleus seemed to disappear, leaving

in its place long thready material that consolidated to form the chromosomal bodies. (See

figure 6a.) Once formed, the chromosomes

assembled along the middle of the cell, copied

themselves, and then moved apart while the cell

pinched itself in half, trapping one set of

chromosomes in each of the two halves. Then

the chromosomes themselves seemed to dissolve as two new nuclei appeared, one in each

of the two newly formed cells. (See figure 6b. )

Thus, the same number of chromosomes appeared in precisely the same form in every cell

of an organism except the germ, or sex, cells.

Furthermore, the chromosomes not only remained constant in form and number from one

generation to the next, but were inherited in

pairs. They were, in short, manifesting all the

traits that Mendel had prescribed for genes

almost three decades earlier. By the beginning

of the 20th century, it was clear that chromosomes were of central importance to the life history of the cell, acting in some unspecified manner as the vehicle for the Mendelian gene.

If this conclusion was strongly implied by the

events of cell division, it became obvious when

reproduction in whole organisms was analyzed.

It had been established by the latter part of the

19th century that the germ cells of plants and

animals—pollen and ovum, sperm and egg—actually fuse in the process of fertilization. Germ

cells differ from other body cells in one important respect—they contain only half the usual

number of chromosomes. This chromosome

halving within the cell was apparently done

very precisely, for every sperm and egg contained exactly one representative from each

chromosome pair. When the two germ cells

then fused during fertilization, the offspring

were supplied with a fully reconstituted chromosome complement, half from each parent.

Clearly, chromosomes were the material link

from one generation to the next. Somewhere

locked within them was the substance of both

heredity—the fidelity of traits between generations; and diversity—the potential for genetic

variation and change.
32 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Figure 6.— Chromosomes

6a. An example of

Step 1

Photo credit: Professor Judith Lengyel, Molecular Biology institute, UCLA

Optical micrograph of chromosomal material from the salivary gland of the larva of the

common fruit fly, Drosophila melanogaster

a chromosome body from a higher organism.

Step 3 Step 4

6b. In Step 1, the chromosome bodies are still uncondensed.

Step 5

Step 6

In Steps 2 and 3, the chromosomes condense into thread-like bodies and align themselves near the center of the cell.

In Steps 4 and 5, the chromosomes begin to separate and are pulled to the opposite poles of the cell.

In Step 6, the chromosomes return to an uncondensed state and the cell begins to constrict about the middle to form

two new cells.

SOURCE: Office of Technology Assessment.
Ch. 2—introduction . 33

(kinetics in the 20th century

During the first few decades of the 20th century, scientists searched for progressively

simpler experimental organisms to clarify progressively more complex genetic concepts. First

was Thomas Hunt Morgan’s Drosophila—gnatsized fruit flies with bulbous eyes. These insects

have a simple array of four easily distinguishable chromosome pairs per cell. They reproduce rapidly and in large numbers under the

simplest of laboratory conditions, supplying a

new generation every month or so. Thus, researchers could carry out an enormous number

of crosses employing a whole catalog of different fruit fly traits in a relatively brief time.

It became obvious from the extensive Drosophila data that certain traits were more likely

to be inherited together than others. Yellow

bodies and ruby eyes, for instance, almost always went together, with both in turn, appearing more frequently than expected with the

trait known as “forked bristles. ” All three traits,

however, showed up only randomly with

curved wings. Certain genes thus seemed to be

linked to one another. The entire Drosophila

genome, in fact, fell into four distinct linkage

groups. The physical basis for these groups, not

surprisingly, consisted of the four fruit fly

chromosomes. Linked genes behaved as they

did because they were located on the same

chromosome.

Soon, scientists learned that they could not

only assign particular genes to particular Drosophila chromosomes but could identify the relative locations of different genes on a given

chromosome. This gene mapping was possible

The riddle of the gene _

because linkage itself was not permanent,

linked genes sometimes separated. For instance,

while yellow bodies, ruby eyes, and forked bristles were all linked traits, the first two stayed

together far more frequently than either did

with the third.

The degree of linkage between two genes was

hypothesized to be directly proportional to the

distance between them on the chromosome,

mainly because of a unique event that occurs

during the development of germ cells. Before

the normal chromosome number is halved, the

chromosomes crowd together in the center of

the cell, coiling tightly around each other, practically fusing along their entire length. It is in

this state that crossing-over (or natural recombination)—the actual physical exchange of parts

between chromosomes—occurs. No chromosome emerges from the exchange in the same

condition as before; the lengths of chromosomes are reshuffled before being transferred

to the next generation.

The idea of linkage meant that Mendel’s formulations had to be modified. Clearly, genes

were not completely independent units. Further

work with Drosophila in the 1920’s showed that

genes were also not permanent and could

change over time. Although natural mutations

occurred at a very slow rate, exposing fruit flies

to X-rays accelerated their frequency enormously. Exposure of a parental fly population

led to an array of new traits among their offspring—traits which, if they were neither lethal

nor sterilizing, could be passed from one generation to the next.

With all this research, nobody yet knew what triguing observations made a decade earlier by

the gene was made of. The first evidence that a British physician, Fred Griffith. He had

it consisted of deoxyribonucleic acid (DNA) worked with two types of pneumococcus (the

emerged from the work of Oswald Avery, Colin bacteria responsible for pneumonia) and with

MacLeod, and Maclyn McCarty at the Rockefel- two different bacteria within each type. One

ler Institute in New York in the early 1940’s. bacterium in each type was coated in a polysacAvery’s group took as its starting point some in- charide capsule; the other was bare. Bare bac-
34 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

teria gave rise only to bare progeny, while those

with capsules produced only encapsulated

forms. Only the encapsulated forms of both

types II and III could cause disease; bare bacteria were benign. (See figure 7a.) But when

Griffith took some encapsulated type III bacteria

that had been killed and rendered harmless and

mixed them with bare bacteria of type II, the

presumably safe mixture became virulent: Mice

injected with it died of a massive pneumonia infection, Bacteria recovered from these animals

were found to be of type 11—the only living bacteria the mice had received—now wrapped in

type III capsules. (See figure 7b.)

Avery’s group recognized Griffith’s finding as

a genetic phenomenon; the dead type 111 bacteria must have delivered the gene for making

capsules into the genetic complement of the

living type 11 recipients. By meticulous research,

Avery’s group found that the substance which

caused the genetic transformation was DNA.

It had been in 1868, just 3 years after Mendel

had published his findings, that DNA was discovered by Friedrich Miescher. It is an extremely simple molecule composed of a small sugar

molecule, a phosphate group (a phosphorous

atom surrounded by four oxygen atoms), and

four kinds of simple organic chemicals known

as nitrogenous (nitrogen-containing) bases. Together, one sugar, one phosphate, and one base

form a nucleotide—the basic structural unit of

the large DNA molecule. Because it is so simple,

DNA had appeared to be little more than a

monotonous conglomeration of simple nucleotides to scientists in the early 20th century. It

seemed unlikely that such a prosaic molecule

could direct the appearance of genetic traits

while faithfully reproducing itself so that information could be transferred between generations. Although Avery’s results seemed clear

enough, many were reluctant to accept them.

Those doubts were finally laid to rest in a

brief report published in 1953 by James Watson

and Francis Crick. By using X-ray crystallographic techniques and building complex models—and without ever having actually seen the

molecule itself—Watson and Crick reported that

they had discovered a consistent scientifically

sound structure for DNA.

Figure 7.—The Griffith Experiment

7a. There are two types of pneumococcus, each of which

can exist in two forms:

where R represents the rough, nonencapsulated, benign

form; and

S represents the smooth, encapsulated, virulent

form.

7b. The experiment consists of four steps:

Virulent strain (1)

Mice injected with nonvirulent Rll do not become infected.

Virulent

strain,

heat-killed

(3)

The virulent SIII is heat-killed. Mice injected with it do not

die.

heat-killed II

(4)

When mice are injected with the nonvirulent RII and the

heat-killed SIII, they die. Type II bacteria wrapped in type Ill

capsules are recovered from these mice.

SOURCE: Office of Technology Assessment.
Ch. 2—introduction . 35

The structure that Crick and Watson uncovered solved part of the genetic puzzle. According to them, the phosphates and sugars formed

two long chains, or backbones, with one nitrogenous base attached to each sugar. The two

backbones were held together like the supports

of a ladder by weak attractions between the

bases protruding from the sugar molecules. Of

the four different nitrogenous bases—adenine,

thymine, guanine, and cytosine—attractions existed only between adenine(A) and thymine(T),

and between guanine(G) and cytosine(C). (See

figure 8a) Thus, if a stretch of nucleotides on

one backbone ran:

the other backbone had to contain the directly

opposite complementary sequence:

T-A-G-A-A-T-T. . .

The complementary pairing between bases running down the center of the long molecule was

responsible for holding together the two otherwise independent chains. (See figure 8b. ) Thus,

the DNA molecule was rather like a zipper, with

the bases as the teeth and the sugar-phosphate

chains as the strands of cloth to which each zipper half was sewn. Crick and Watson also found

that in the presence of water, the two poly -

nucleotide chains did not stretch out to full

length, but twisted around each other, forming

what has undoubtedly become the most glorified structure in the history of biology-the double helix. (See figure 8c.)

The structure was scientifically elegant. But it

was received enthusiastically also because it implied how DNA worked. As Crick and Watson

themselves noted:

If the actual order of the bases on one of the

pair of chains were given, one could write down

the exact order of the bases on the other one,

because of the specific pairing. Thus one chain

is, as it were, the complement of the other, and

it is this feature which suggests how the desoxyribonucleic acid molecule might duplicate

itself. ’

When a double-stranded DNA molecule is unzipped, it consists of two separate nucleotide

chains, each with a long stretch of unpaired

bases. In the presence of a mixture of nucleotides, each base attracts its complementary

match in accordance with the inherent affinities

of adenine for thymine, thymine for adenine,

guanine for cytosine, and cytosine for guanine.

The result of this replication is two DNA molecules, both precisely identical to each other and

to the original molecule—which explains the

faithful duplication of the gene for passage from

one generation to the next. (See figure 9.)

Crick and Watson’s work solved a major riddle in genetic research. Because George Beadle

and Edward Tat urn had recently discovered

that genes control the appearance of specific

proteins, and that one gene is responsible for

producing one specific protein, scientists now

knew what the genetic material was, how it replicated, and what it produced. But they had yet

to determine how genes expressed themselves

and produced proteins.

‘James D. Watson and Francis Crick, %enetical Implications of

the Structures of Deoxyribose Nucleic Acid, ” Nature 171, 1953. pp.

737-8.
36 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

A

Adenine

Figure 8.—The Structure of DNA

o

-o”

5

0 .

-o”

8a. The pairing of the four nitrogenous bases of DNA:

Adenine (A) pairs with Thymidine (T)

Guanine (G) pairs with Cytosine (C) -o”

o .

8b. The four bases form the four letters in the alphabet of

the genetic code. The sequence of the bases along the

sugar-phosphate backbone encodes the genetic information.

A schematic diagram of the DNA double helix. A three-dimensional representation of the DNA double helix.

&. The DNA molecule is a double helix composed of two chains. The sugar-phosphate backbones twist around the outside, with the paired bases on the inside serving to hold the chains together.

SOURCE: Office of Technology Assessment.
Ch. 2—introduction . 37

The genetic

Figure 9.—Replication of DNA

Old Old

Old New New Old

When DNA replicates, the original strands unwind and

serve as templates for the building of new complementary

strands. The daughter molecules are exact copies of the

parent, with each having one of the parent strands.

SOURCE: Office of Technology Assessment.

code

Proteins are the basic materials of cells. Some Ironically, proteins are far more complex and

proteins are enzymes, which catalyze reactions diverse than the four nucleotides that help

within a cell. In general, for every chemical re- create them. Proteins, too, are long chains made

action in a living organism, a specific enzyme is up of small units strung together. In this case,

required to trigger the process. Other proteins however, the units are amino acids rather than

are structural, comprising most of the raw ma- nucleotides—and there are 20 different kinds of

terial that forms cells. amino acids. Since an average protein is a few

76-565 0 - 81 - 4
38 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

hundred amino acids in length, and since any

one of 20 amino acids can fill each slot, the numher of possible proteins is enormous. Nevertheless, each protein requires the strictest ordering

of amino acids in its structure. Changing a

single amino acid in the entire sequence can

drastically change the protein’s character.

It was now possible for scientists to move

nearer to an appreciation of how genes functioned. First had come the recognition that DNA

determined protein; now it was evident that the

sequence of nucleotides in DNA determined a

linear sequence of amino acids in proteins.

By the early 1980’s, the way proteins were

manufactured, how their synthesis was regulated, and the role of DNA in both processes

were understood in considerable detail. The

process of transcribing DNA’s message-carrying the message to the cell’s miniature protein

factories and building proteins-took place

through a complex set of reactions. Each amino

acid in the protein chain was represented by

three nucleotides from the DNA. That threebase unit acted as a word in a DNA sentence

that spelled out each protein-the genetic code.

(See figure 10.)

Through the genetic code, an entire gene—a

linear assemblage of nucleotides-could now be

THIRD BASE

Figure 10.—The Genetic Code

THIRD BASE

SECOND THIRD BASE THIRD BASE

● och (ochre); amb (ambert, and end are step signals for translation, i.e.,

signals the end of synthesis of the protein chain.

Amino acid

alanine

arginine

asparagine

aspartic acid

asn and/or asp

cysteine

glutamine

glutamic acid

gin and/or glu

glycine

histidine

isoleucine

Ieucine

Iysine

methionine

phenylaianine

proline

serin e

threonine

tryptophan

tyrosine

valine

symbol

ala

arg

asn

asp

asx

Cys

gln

glu

glx

gly

his

ileu

Ieu

Iys

met

phe

pro

ser

thr

trp

tyr

val

Each amino acid is determined by a

three letter code (A, G, T, or C) along

the DNA. If the first letter in the code

is A, the second is T, and third is A,

the amino acid will be tyrosine (or tyr)

in the complete protein molecule. For

Ieucine (or Ieu), the code is GAT, and

so forth. The dictionary above gives

the entire code,

SOURCE: Office of Technology Assessment.
Ch. 2—introduction ● 3 9

read like a book. By the 1970’s, researchers had Figure 11.—The Expression of Genetic Information

learned to read the code of certain proteins, in the Cell

synthesize their DNA, and insert the DNA into

bacteria so that the protein could be produced.

(See figure 11.)

Meanwhile, other scientists were studying

the genetics of viruses and bacteria. The combination of these studies with those investigating the genetic code led to the innovations of

genetic engineering.

Developing

In the early 1960’s,

SOURCE: Office of Technology Assessment.

genetic technologies

scientists discovered ex- DNA along with it. Thus, when the new virus

actly how genes move from one bacterium to

another. One such mechanism uses bacteriophages-viruses that infect bacteria-as intermediaries. Phages act like hypodermic needles,

injecting their DNA into bacterial hosts, where

it resides before being passed along from one

generation to the next as part of the bacterium’s

own DNA. Sometimes, however, the injected

DNA enters an active phase and produces a crop

of new virus particles that can then burst out of

their host. Often during this process, the viral

DNA inadvertently takes a piece of the bacterial

particles now infect other bacteria, they bring

along several genes from their previous host.

This viral transduction-the transfer of genes

by an intermediate viral vector or vehiclecould be used to confer new genetic traits on

recipient bacteria. (See figure 12. )

Bacteria also transfer genes directly in a process called ’conjugation, in which one bacterium

attaches small projections to the surface of a

nearby bacterium. DNA from the donor bacterium is then passed to the recipient through the

Figure 12.—Transduction: The Transfer of Genetic Material in Bacteria by Means of Viruses

Bacterium

Bacterial

Empty chromosome

viral coat fragments

. New virus

bacterium

In step 1 of viral transduction, the infecting virus injects its DNA into the cell. In step 2 when the new viral particles are

formed, some of the bacterial chromosomal fragments, such as gene A, may be accidently incorporated into these progeny

viruses instead of the viral DNA. In step 3 when these particles infect a new cell, the genetic elements incorporated from the

first bacterium can recombine with homologous segments in the second, thus exchanging gene A for gene a.

SOURCE: Office of Technology Assessment
40 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

projections. The ability to form projections and

donate genes to neighbors is a genetically controlled trait. The genes controlling this trait,

however, are not located on the bacterial chromosomes. Instead, they are located on separate

genetic elements called plasmids-relatively

small molecules of double-stranded DNA, arranged as closed circles and existing autonomously within the bacterial cytoplasm. (See

figure 13.)

Plasmids and phages are two vehicles—or

vectors-for carrying genes into bacteria. As

such, they became tools of genetic engineering;

for if a specifically selected DNA could be introduced into these vectors, it would then be possible to transfer into bacteria the blueprints for

proteins—the building blocks of genetic characteristics.

But bacteria had been confronting the invasion of foreign DNA for millennia, and they had

evolved protective mechanisms that preserved

their own DNA while destroying the DNA that

did not belong. Bacteria survive by producing

restriction enzymes. These cut DNA molecules

in places where specific sequences of nucleotides occur—snipping the foreign DNA, yet leaving the bacteria’s own genetic complement

alone. The first restriction enzyme that was isolated, for instance, would cut DNA only when it

located the sequence:

(G-A-A-)

(C-T-T)

If the sequence occurred once in a circular plasmid, the effect would simply be to open the

circle. If the sequence were repeated several

times along a length of DNA, the DNA would be

chopped into several small pieces.

By the late 1970’s, scores of different restriction enzymes had been isolated from a variety

of bacteria, with each enzyme having a unique

specificity for one specific nucleotide sequence.

These enzymes were another key to genetic engineering: they not only allowed plasmids to be

opened up so that new DNA could be inserted,

but offered a way of obtaining manageable

pieces of new DNA as well. (See figure 14.)

Using restriction enzymes, almost any DNA

molecule could be snipped, shaped, and

trimmed with precision.

Cloning DNA—that is obtaining a large quantity of exact copies of any chosen DNA molecule

by inserting it into a host bacterium-became

technically almost simple. The piece in question

was merely snipped from the original molecule,

inserted into the vector DNA, and provided with

Figure 13.—Conjugation: The Transfer of Genetic Material in Bacteria by Mating

Plasmid

In conjugation, a plasmid inhabiting a bacterium can transfer the bacterial chromosome to a second cell where homologous

segments of DNA can recombine, thus exchanging gene B from the first bacterium for gene b from the second.

SOURCE: Office of Technology Assessment.
Ch. 2—introduction ● 4 1

Figure 14.—Recombinant DNA: The Technique of

Recombining Genes From One Species

With Those From Another

New DNA

amount of DNA protein

Restriction enzymes recognize certain sites along the DNA

and can chemically cut the DNA at those sites. This makes

it possible to remove selected genes from donor DNA molecules and insert them into plasmid DNA molecules to form

the recombinant DNA. This recombinant DNA can then be

cloned in its bacterial host and large amounts of a desired

protein can be produced.

SOURCE: Office of Technology Assessment.

a bacterial host as a suitable environment for

replication. The desired piece of DNA could be

recombined with a plasmid vector, a procedure

that gave rise to recombinant DNA (rDNA), aIso

known as gene splicing. Since bacteria can be

grown in vast quantities, this process could

result in large-scale production of otherwise

scarce and expensive proteins.

Although placing genes inside of bacteria is

now a relatively straightforward procedure, obtaining precisely the right gene can be difficult.

Three techniques are currently available:

● Ribonucleic acid—RNA—is the vehicle

through which the message of DNA is read

and transcribed to form proteins. The RNA

that carries the message for the desired

protein is first isolated. An enzyme, called

‘reverse transcriptase, is then added to the

RNA. The enzyme triggers the formation of

DNA—reversing the normal process of protein production. The DNA is then inserted

●

●

into an appropriate vector. This was the

procedure used to obtain the gene for human insulin in 1979. (See figure 15. )

The gene can also be synthesized, or

created, directly, since the nucleotide sequence of the gene can be deduced from

the amino acid sequence of its protein

product. This procedure has worked well

for small proteins—like the growth regulatory hormone somatostatin—which have

relatively short stretches of DNA coding.

But somatostatin is a tiny protein, only 14

amino acids long. With three nucleotides

coding for each amino acid, scientists had

to synthesize a DNA chain 42 nucleotides

long to produce the complete hormone. For

larger proteins, the gene-synthesis approach rapidly becomes highly impractical.

The third method is also the most controversial. In this “shotgun” approach, the

entire genetic complement of a cell is

chopped up by restriction enzymes. Each

of the DNA fragments is attached next to

vectors and transferred into a bacterium;

the bacteria are then screened to find those

making the desired product. Screening

thousands of bacterial cultures was part of

the technique that enabled the isolation of

the human interferon gene. *

At present, these techniques of recombination work mainly with simple micro-organisms.

Scientists have only recently learned how to introduce novel genetic material into cells of

higher plants and animals. These higher cells

are being ‘engineered’ in totally different ways,

by growing plant or animal cells in ‘tissue culture’ systems, in vitro.

Tissue culture systems work with isolated

cells, with entire pieces of tissue, and to a far

more limited extent, with whole organs or even

early embryos. The techniques make it possible

to manipulate cells experimentally and under

controlled conditions. Several techniques are

available. For example, in one set of experiments, complete plants have been grown from

single cells—a breakthrough that may permit

“Strictly speaking, RNA was transcribed using the shotgun

approach into DNA, which was then cloned into bacteria and

screened.
42 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Figure 15.-An Example of How the Recombinant DNA Technique May Be Used

To Insert New Genes Into Bacterial Cells

The first part of the technique involves the manipula- 11. A bacterial plasmid, which is a small piece of circular

tions necessary to isolate and reconstruct the desired DNA, serves as the vehicle for introducing the new gene

gene from the donor: (obtained in part I above) into the bacterium:

‘a)

b)

c)

The RNA that carries the message (mRNA) for the

. . ,

a) The circular plasmid is cleaved by the appropriate

desired protein product is isolated. restriction enzyme.

The double-stranded DNA is reconstructed from the b) The enzyme terminal transerase extends the DNA

mRNA. strands of the broken circle with identical bases

in the final step of this sequence, the enzyme ter- (four cytosines in this case, to allow complemenminal transferase acts to extend the ends of the

DNA strands with short sequences of identical

bases (in this case four guanines).

I

Messenger RNA

from animal cell

a)

Enzymatic

reconstruction

Double-strand DNA t

b)

I

Terminal

transferase

Ill. The final product, a bacterial plasmid containing the

new gene, is obtained. This plasmid can then be inserted into a bacterium where it can be replicated and

produce the desired protein product:

a)

b)

tary base pairing with the guanines added to the

gene obtained in part i).

The gene obtained in part i and the plasmid DNA

from part ii are mixed together and anneai

because of the complementary base-pairing between them.

Bacterial enzymes fill in any gaps in the circle,

sealing the connection between the plasmid DNA

and the inserted DNA to generate an intact circular plasmid now containing a new gene.

Uptake by cell;

Ill repair by

II

Bacterial plasmid DNA

a)

enzymes

SOURCE: Office of Technology Assessment.
Ch. 2—introduction ● 4 3

hundreds of plants to be grown asexually from

a small sample of plant material. Just as with

bacteria, the cells can be induced to take up

pieces of DNA in a process call transformation.

They can also be exposed to mutation-causing

agents so that they produce mutants with desired properties. in another set of experiments,

two different cells have been fused to form a

new, single-cell “hybrid” that contains the

genetic complements of both antecedents. In

both cases, the success of tissue culture and cell

The basic issues

Applied genetics is like no other

By itself, it may enable tremendous

fusion* can be used to direct efficient, fast genetic changes in plants. (See ch. 8.)

Cell culture techniques, while not strictly genetic manipulation, form a major aspect of modern biotechnology. Combined with genetic approaches, their potential is only on the verge of

being realized.

● A related technique is protoplasm fusion, or the fusion of cells

whose walls have been removed to leave only membrane-bound

cells. The cells of bacteria, fungi, and plants must all be freed of

their walls before they can be fused.

technology.

advances in

conquering diseases, increasing food production, producing new and cheaper industrial substances, cleaning up pollution, and understanding the fundamental processes of life. Because

the technology is so powerful, and because it involves the basic roots of life itself, it carries with

it potential hazards, some of which might arise

from basic research, others of which may stem

from its applications.

As the impacts of genetic technologies are discussed, two fundamental questions must be

kept in mind:

How will applied genetics be used?

Interest in the industrial use of biological

processes stems from a merging of two paths:

the revolution in scientific understanding of the

nature of genetics; and the accelerated search

for a sustainable society in which most industrial processes are based on the use of renewable resources. The new genetic technologies

will spur that search in three ways: they will

provide a means of doing something biologically—with renewable raw materials—that previously required chemical processes using nonrenewable resources; they will offer more efficient, more economical, less polluting ways for

producing both old and new products; and they

will increase the yield of the plant and animal

resources that are responsible for providing the

world’s supplies of food, fibers, and some fuels.

What are the dangers?

Even before scientists recognized the potential power of applied genetics, some questioned

its consequences; for with its benefits, appeared

hypothetical risks. Although most experts today

agree that the immediate hazards of the basic

research itself appear to be minimal, nobody

can be certain about all the consequences of

placing genetic characteristics in micro-organisms, plants, and animals that have never carried them before. There are at least three separate areas of concern:

First, genetically engineered micro-organisms

might have potentially deleterious effects on human health, other living organisms, or the environment in general. Unlike toxic chemicals, organisms may reproduce and spread of their

own accord; if they are released into the envi -

ronment, they may be impossible to control.

Second, some observers question whether

sufficient knowledge exists to allow the extinction of diverse species of “genetically inferior”

plants and animals in favor of a few strains of

“superior” ones. Evolution thus far has depended, in part, on genetic diversity; replacing

in nature diverse inferior strains by genetically

engineered superior strains may increase the

susceptibility of living things to disease and environmental insults.

Finally, this new knowledge affects the understanding of life itself. It is tied to the ultimate
44 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

questions of how humans view themselves and locating and monitoring its benefits and burwhat they legitimately control in the world. dens. That process requires knowledge. The following sections of the report describe the im- Because of the significant and wide-ranging

scope of applied genetics, society as whole must pacts of applied genetics on specific industries,

begin to debate the issues with a view toward al- and assess many of their consequences.
Part I

Biotechnology

Chapter 3-Genetic Engineering in the Fermentation Technologies . . . . . . , . . . . . . . . . 49

Chapter 4-The Pharmaceutical Industry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59

Chapter 5-The Chemical Industry. . . . . . . . . . . . . , . . . . . . . . . . . . . . . . . . . , . . . . . . . 85

Chapter 6-The Food Processing Industry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107

(C hapter 7- The Use of Genetically Engineered Micro-Organisms in the Environment. . 117
Chapter 3

Genetic Engineering and the

Fermentation Technologies
Chapter 3

Page

Biotechnology—An Introduction. . . . . . . . . . . . . 49

Fermentation . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

Fermentation Industries . . . . . . . . . , . . . . . . . 50

Fermentation Using Whole Living Cells . . . . . . 51

The Process of Enzyme Technology. . . . . . . . . 53

Comparative Advantages of Fermentations

Using whole Cells and Isolated Enzymes. . . . 54

The Relationship of Genetics to Fermentation . 54

Fermentation and Industry . . . . . . . . . . . . . , . 55

Table

Figures

Figure No.

16. Diagram of Products Available From Cells . . . 51

17. Features of a Standard Fermenter . . . . . . . . . 52

18. Immobilized Cell System . . . . . . . . . . . . . . . . 52

19. Diagram of Conversion of Raw Material

to Product. . . . . . . . . . . . . . . . . . . . . . . . . .

Table No. Page

2. Enzyme Products . . . . . . . . . . . . . . . . . . . . . . 54
Chapter 3

Genetic Engineering and

the Fermentation Technologies

Biotechnology—an introduction

Biotechnology involves the use in industry of

living organisms or their components (such as

enzymes). It includes the introduction of genetically engineered micro-organisms into a variety

of industrial processes.

The pharmaceutical, chemical, and food processing industries, in that order, are most likely

to take advantage of advances in molecular genetics. Others that might also be affected, although not as immediately, are the mining,

crude oil recovery, and pollution control industries.

Because nearly all the products of biotechnology are manufactured by micro-organisms, fermentation is an indispensable element of biotechnology’s support system. The pharmaceutical industry, the earliest beneficiary of the new

knowledge, is already producing pharmaceuticals derived from genetically engineered

micro-organisms. The chemical industry will

take longer to make use of biotechnology, but

the ultimate impact may be enormous. The food

processing industry will probably be affected

last.

This report examines many of the pharmaceutical industry’s products in detail, as well as

Fermentation

There are several ways that DNA can be cut,

spliced, or otherwise altered. But engineered

DNA by itself is a static molecule. To be anything more than the end of a laboratory exercise, the molecule must be integrated into a system of production; to have an impact on society

at large, it must become a component of an industrial or otherwise useful process.

The process that is central to the economic

some of the secondary impacts that the technologies might have. Because the chemical and

food industries will feel the major impact of biotechnology later, specific impacts are less certain and particular products are less identifiable. The mining, oil recovery, and pollution

control industries are also candidates for the

use of genetic technologies. However, because

of technical, scientific, legal, and economic uncertainties, the success of applications in these

industries is more speculative.

The generalizations made with respect to

each of the industries should be viewed as just

that—generalizations. Because a wide array of

products can be made biologically, and because

different factors influence each instance of production, isolated examples of success may appear throughout the industries at approximately the same time. In almost every case, specific

predictions can only be made on a product-byproduct basis; for while it may be true that biotechnology’s overall impact will be profound,

identifying many of the products most likely to

be affected remains speculative.

success of biotechnology has been around for

centuries. It is fermentation, essentially the

process used to make wine and beer. It can also

produce organic chemical compounds using

micro-organisms or their enzymes.

Over the years, the scope and efficiency of

the fermentation process has been gradually improved and refined. Two processes now exist,

both of which will benefit from genetic engi49
50 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

neering. In fermentation technology, living organisms serve as miniature factories, converting raw materials into end products. In enzyme

technology, biological catalysts extracted from

those living organisms are used to make the

products.

Fermentation industries

The food processing, chemical, and pharmaceutical industries are the three major users of

fermentation today. The food industry was the

first to exploit micro-organisms to produce

alcoholic beverages and fermented foods. Mid16th century records describe highly sophisticated methods of fermentation technology. Heat

processing techniques, for example, anticipated

pasteurization by several centuries.

[n the early 20th century, the chemical industry began to use the technology to produce

organic solvents like ethanol, and enzymes like

amylase, used at the time to treat textiles. The

chemical industry’s interest in fermentation

arose as the field of biochemistry took shape

around the turn of the century. But it was not

until World War I that wartime needs for the

organic solvent acetone—to produce the cordite used in explosives–substantially increased

research into the potential of fermentation.

Thirty years later after World War 11, the pharmaceutical industry followed the chemical industry’s lead, applying fermentation to the production of vitamins and new antibiotics.

Today, approximately 200 companies in the

United States and over 500 worldwide use

fermentation technologies to produce a wide

variety of products. Most use them as part of

production processes, usually in food processing. But others manufacture either proteins,

which can be considered primary products, or a

host of secondary products, which these proteins help produce. For genes can make enzymes, which are proteins; and the enzymes

can help make alcohol, methane, antibiotics,

and many other substances.

Proteins, the primary products, function as:

 enzymes such as asparaginase which are

used in the treatment of leukemia;

●

●

●

●

structural components, such as collagen,

used in skin transplants following burn

trauma;

certain hormones, such as insulin and

human growth hormone;

substances in the immune system, such as

antibodies and interferon; and

specialized functional components, such as

hemoglobin.

Fermentation technologies are so useful for producing proteins partly because these are the

direct products of genes. But proteins (as enzymes) can also be used in thousands of additional conversions to produce practically any

organic chemical and many inorganic ones as

well: (See figure 16. )

Figure 16.-Diagram of Products Available

From Cells

isolated

B

Raw material Product

In (A) DNA directs the formation of a protein, such as insulin, which is itself the desired product. In (B), DNA directs

the formation of an enzyme which, in turn, converts some

raw material, such as sugar, to a product, such as ethanol.

SOURCE: Office of Technology Assessment.
Ch. 3—Genetic Engineering and the Fermentation Technologies . 51

carbohydrates, such as fructose sweeteners;

lipids, such as vitamins A, E, and K;

alcohols, such as ethanol;

other organic compounds, such as acetone;

and

inorganic chemicals, such as ammonia, for

use in fertilizers.

Fermentation is not the only way to manufacture or isolate these products. Some are traditionally produced by other methods. If a change

from one process to another is to occur, both

economic and societal pressures will help determine whether an innovative approach will be

used to produce a particular product. Alan Bull

has identified four stimuli for change and innovation: 1

1. abundance of a potentially useful raw

material;

2. scarcity of an established product;

3. discovery of a new product; and

4. environmental concerns.

And conditions existing today have added a fifth

stimulus:

5. scarcity of a currently used raw material.

Each of these factors has tended to accelerate

the application of fermentation.

1. Abundance of a potentially useful raw material.—The use of a raw material can be

the driving force in developing a process.

When straight chain hydrocarbons (n-alkanes) were produced on a large scale as

petroleum refinery byproducts, fermentation processes were developed to convert

them to single-cell proteins for use in animal feed.

2. Scarcity of an established product.—The

new-found potential for producing human

hormones through fermentation technology is a major impetus to the industry today. Similarly, many organic compounds

once obtained by other processes—like

citric acid, which was extracted directly

‘/\. ‘I”. lhJll, 11. [:. FllWrCM)Ci, and (:. Rat ledge, kficrobia/ Techndo,gv:

(~urrent Sfafe, Future Prospects, Z$lth Symposium ot’ the Society for

(kllf>l’iil hlicrut)iub~y al 1 llll)f~rsitv uf (:iilllt)l’itl~f~, April 1979

((:illllt)l’idg(’, l’;ll~lillld: (kmt)ridge [~l;ilwrsit}~ Press, 1 979), pp. 4-8.

3.

4.

5.

from citrus fruits—are now made by fermentation. As a result of more efficient

technology, products from vitamin B12 to

steroids have come into wider use.

Discovery of a new product.—The discovery

that antibiotics were produced by microorganisms sparked searches for an entirely

new group of products. Several thousand

antibiotics have been discovered to date, of

which over a hundred have proved to be

clinically useful.

Environmental concerns.—The problems of

sewage treatment and the need for new

sources of energy have triggered a search

for methods to convert sewage and municipal wastes to methane, the principal component of natural gas. Because micro-organisms play a major role in the natural cycling of organic compounds, fermentation

has been one method used for the conversion.

Scarcity of a currently used raw materia/.—Because the Earth’s supplies of fossil

fuels are rapidly dwindling, there is intense

interest in finding methods for converting

other raw materials to fuel. Fermentation

offers a major approach to such conversions.

Fermentation technologies can be effective in

each of these situations because of their outstanding versatility and relative simplicity. The

processes of fermentation are basically identical, no matter what organism is selected, what

medium used, or what product formed. The

same apparatus, with minor modifications, can

be used to produce a drug, an agricultural product, a chemical, or an animal feed supplement.

Fermentation using whole living cells

Originally, fermentation used some of the

most primitive forms of plant life as cell factories. Bacteria were used to make yogurt and

antibiotics, yeasts to ferment wine, and the

filamentous fungi or molds to produce organic

acids. More recently, fermentation technology

has begun to use cells derived from higher

plants and animals under growth conditions

known as cell or tissue culture. In all cases,

large quantities of cells with uniform character-
52 . Impacts of Applied Genetics —Micro-Organisms, Plants, and Animals

istics are grown under defined, controlled conditions.

In its simplest form, fermentation consists of

mixing a micro-organism with a liquid broth

and allowing the components to react. More sophisticated large-scale processes require control

of the entire environment so that fermentation

proceeds efficiently and, more importantly, so

that it can be repeated exactly, with the same

amounts of raw materials, broth, and microorganisms producing the same amount of product. Strict control is maintained of such variables as pH (acidity/alkalinity), temperature, and

oxygen supply. (See figure l?. ) The newest models are regulated by sensors that are monitored

by computers. The capacity of industrial-sized

fermenters can reach 50,000 gal or more. The

one-shot system of fermentation is called batch

fermentation—i.e., fermentation in which a

single batch of material is processed from start

to finish.

In continuous fermentation, an improvement

on the batch process, fermentation goes on

without interruption, with a constant input of

Figure 17.- Features of a Standard Fermenter

Exhaust

to atmosphere

unfiltered

Compressed air harvest &

inlet

sampler

fill

Cooling jacket

Agitator

Air sparger

raw materials and other nutrients and an attendant output of fermented material. The most

recent approaches use micro-organisms that

have been immobilized in a supporting structure. (See figure 18.) As the solution containing

the raw material passes over the cells, the

micro-organisms process the material and release the products into the solution flowing out

of the fermenter.

In general, products obtained by fermentation also can be produced by chemical synthesis, and to a lesser extent can be isolated by extraction from whole organs or organisms. A

fermentation process is usually most competitive when the chemical process requires several

Figure 18.-immobilized Cell System

Solution with product out

.

anisms

u immobilized in inert

material

Raw material solution in

Typically, a solution of raw materials is pumped through a

bed of immobilized micro-organisms which convert the

materials to the desired product.

SOURCE: Eli Lilly & Co. SOURCE: Office of Technology Assessment.
Ch. 3—Genetic Engineering and the Fermentation Technologies ● 53

individual steps to complete the conversion. In a

chemical synthesis, the raw material (shown in

figure 19 as a) might have to be transformed to

an intermediate b, which, in turn, might have to

be converted to intermediates c and d before

final conversion to the product e–each step

necessitating the recovery of its products before

the next conversion. In fermentation technology, all steps take place within those miniature

chemical factories, the micro-organisms; the

microbial chemist merely adds the raw material

a and recovers the product e.

A wide variety of carbohydrate raw materials

can be used in fermentation. These can be pure

substances (sucrose or table sugar, glucose, or

fructose) or complex mixtures still in their

original form (cornstalks, potato mash, sugarcane, sugar beets, orcellulose). They can be of

recent biological origin (biomass) or derived

from fossil fuels (methane or oil). The availability of raw materials varies from country to coun3) In the Chemical Conversion of raw material a to final

product e, intermediates b, c, and d must be synthesized. Each intermediate must be recovered and purified

before it can be used in the next step of the conversion.

)) A cell can perform the same conversion of a to e, but

with the advantage that the chemist does not have to

deal with the intermediates: the raw material a k simply

added and the final product e, recovered.

OURCE: Office of Technology Assessment.

try and even from region to region within a

country; the economics of the production process varies accordingly.

The cost of the raw material can contribute

significantly to the cost of production. Usually,

the most useful micro-organisms are those that

consume readily available inexpensive raw materials. For large volume, low-priced products

(such as commodity chemicals), the relationship

between the cost of the raw material and the

cost of the end product is significant. For low

volume, high-priced products (such as certain

pharmaceuticals), the relationship is negligible.

The process of enzyme technology

Although live yeast had been used for several

thousand years in the production of fermented

foods and beverages, it was not until 1878 that

the active agents of the fermentation process

were given the name “enzymes” (from the

Greek, meaning “in yeast”). The inanimate

nature of enzymes was demonstrated less than

two decades later when it was shown that extracts from yeast cells could effect the conversion of glucose to ethanol. Finally, their actual

chemical nature was established in 1926 with

the purification and crystallization of the

enzyme urease.

Fermentation carried out by live cells provided the conceptual basis for designing fermentation processes based on isolated enzymes.

A single enzyme situated within a living cell is

needed to convert a raw material into a product. A lactose-fermenting organism, e.g., can be

used to convert the sugar lactose, which is

found in milk, to glucose (and galactose). But if

the actual enzyme responsible for the conversion is identified, it can be extracted from the

cell and used in place of a living cell. The

purified enzyme carries out the same conversion as the cell, breaking down the raw material

in the absence of any viable micro-organism. An

enzyme that acts inside a cell to convert a raw

material to a product can also do this outside of

the cell.

Both batch and continuous methods are used

in enzyme technology. However, in the batch

method, the enzymes cannot be recovered eco-
—

54 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals .

nomically, and new enzymes must be added for

each production cycle. Furthermore, the enzymes are difficult to separate from the end

product and constitute a potential contaminant.

Because enzymes used in the continuous method are reusable and tend not to be found in the

product, the continuous method is the method

of choice for most processes. Depending on the

desired conversion, the immobilized microorganisms of figure 18 could be replaced by an

appropriate immobilized enzyme.

Although more than 2,000 enzymes have

been discovered, fewer than 50 are currently of

industrial importance. Nevertheless, two major

features of enzymes make them so desirable:

their specificity and their ability to operate

under relatively mild conditions of temperature

and pressure. (The most frequently used. enzymes are listed in table 2.)

Comparative advantages of

fermentations using whole cells

and isolated enzymes

At present, it is still uncertain whether the

use of whole cells or isolated enzymes will be

more useful in the long run. There are advantages and disadvantages to each. The role of genetic engineering in the future of the industry,

Table 2.—Enzyme Products

Commercially Current available before: production

Source/name 1900 1950 1980 tons/yr

Animal

Rennet. . . . . . . . . . . . . . . X 2

Trypsin. . . . . . . . . . . . . . . x 15

Pepsin . . . . . . . . . . . . . . . x 5

Plant

Malt amylase. . . . . . . . . . X 10,000

Papain . . . . . . . . . . . . . . . x 100

Microbial

Koji. . . . . . . . . . . . . . . . . . X

Fungal protease . . . . . . . X 10

Bacillus protease . . . . . . x 500

Amyloglucosidase . . . . . x 300

Fungal amylase. . . . . . . . X 10

Bacterial amylase. . . . . . x 300

Pectinase. . . . . . . . . . . . . x 10

Glucose isomerase. . . . . x 50

Microbial rennet . . . . . . . x 10

SOURCE: Office of Technology Assessment.

however, will be partly determined by which

method is chosen. With isolated enzymes, genetic manipulation can readily increase the supply of enzymes, while with whole organisms, a

wide variety of manipulations is possible in constructing more productive strains.

The relationship of genetics

to fermentation

Applied genetics is intimately tied to fermentation technology, since finding a suitable species of micro-organism is usually the first step in

developing a fermentation technique. Until recently, geneticists have had to search for an

organism that already produced the needed

product. However, through genetic manipulation a totally new capability can be engineered;

micro-organisms can be made to produce substances beyond their natural capacities. The

most striking successes have been in the pharmaceutical industry, where human genes have

been transferred to bacteria to produce insulin,

growth hormone, interferon, thymosin a-1, and

somatostatin, (See ch. 4.)

In general, once a species is found, conventional methods have been used to induce mutations that can produce even more of the desired

compound. The geneticist searches from among

hundreds of mutants for the one micro-organism that produces most efficiently. Most of the

many methods at the microbiologist’s disposal

involve trial-and-error. Newer genetic tech.

nologies, such as the use of recombinant DNA

(rDNA), allow approaches in which useful genet

ic traits can be inserted directly into the micro

organism.

The current industrial approach to fermenta

tion technologies therefore considers two prob

lems: First, whether a biological process car

produce a particular product; and second, wha

micro-organism has the greatest potential for

production and how the desired characteristic;

can be engineered for it. Finding the desired

micro-organism and improving its capability i

so fundamental to the fermentation industry

that geneticists have become important mem

bers of fermentation research teams.
Ch. 3—Genetic Engineering and the Fermentation Technologies ● 55

Genetic engineering can increase an organism’s productive capability (a change that can

make a process economically competitive); but it

can also be used to construct strains with characteristics other than higher productivity. Properties such as objectionable color, odor, or slime

can be removed. The formation of spores that

could lead to airborne spread of the microorganism can be suppressed. The formation of

harmful byproducts can be eliminated or reduced. Other properties, such as resistance to

bacterial viruses and increased genetic stability,

can be given to micro-organisms that lack them.

Applying recent genetic engineering techniques to the production of industrially valuable

enzymes may also prove useful in the future.

For example, a strain of micro-organism that

carries the genes for a desired enzyme may be

pathogenic. If the genes that express (produce)

the enzyme can be transferred to an innocuous

micro-organism, the enzyme can be produced

safely.

CURRENT TECHNICAL LIMITS ON

GENETIC ENGINEERING

Despite the many genetic manipulations that

are theoretically possible, there are several

notable technical limitations:

●

●

●

Genetic maps—the identification of the location of desired genes on various chromosomes have not been constructed for most

industrially useful micro-organisms.

Genetic systems for industrially useful

micro-organisms, such as the availability of

useful vectors, are at an early stage of

development.

Physiological pathways—the sequence of

enzymatic steps leading from a raw material to the desired product, are not known

for many chemicals. Much basic research

will be necessary to identify all the steps.

The number of genes necessary for the conversion is a major limitation. Currently,

rDNA is most useful when only a single

easily identifiable gene is needed. It is more

difficult to use when several genes must be

transferred. Finally, the problems are formidable, if not impossible, when the genes

have not yet been identified. This is the

case with many traits of agronomic importance, such as plant height.

Even if the genes are identified and successfully transferred, methods must be developed to recognize the bacteria that received them. Therefore, the need to develop appropriate selection methods has impeded the application of molecular genetics.

As a consequence of these limitations, genetic

engineering will be applied to the development

of capabilities that require the transfer of only

one or a few identified genes.

Fermentation and industry

Genetic engineering is not in itself an industry, but a technology used at the laboratory

level. It allows the researcher to alter the hereditary apparatus of a living cell so that the cell

can produce more or different chemicals, or

perform completely new functions. The altered

cell, or more appropriately the population of

altered identical cells is, in turn, used in industrial production. It is within this framework that

the impacts of applied genetics in the various industries is examined.

Regardless of the industry, the same three

criteria must be met before genetic technologies

can become commercially feasible. These criteria represent major constraints that industry

must overcome before genetic engineering can

play a part in bringing a product to market.

They include the need for:

1. a useful biochemical product;

2. a useful biological fermentation approach

to commercial production; and

3. a useful genetic approach to increase the

efficiency of production.

The three criteria interrelate and can be met

in any order; the demonstration of usefulness

can begin with any of the three. Insulin, e.g.,

was first found to have value in therapy;

fermentation was then shown to be useful in

its production; and, now genetic engineering

promises to make the fermentation process economically competitive. In contrast, the value of

thymosin a-1, has not yet been proved, although
. Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

the usefulness of genetic engineering and fer- fulness of genetic technologies must be proved.

mentation in its production have been demon- In others, the genetic technologies make prostrated. duction at the industrial level possible, but their

As these examples indicate, the limits on a market has not yet been established. In still

product’s commercial potential vary with the others, the feasibility of fermentation is the maproduct. In some cases, the usefulness of the jor problem.

product has already been shown, and the use-
Chapter 4

The Pharmaceutical Industry
Chapter 4

Page

Background . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59

Past Uses of Genetics. . . . . . . . . . . . . . . . . . . . 59

Potential Uses of Molecular Genetic Technologies 61

Hormones . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61

Insulin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65

Growth Hormone . . . . . . . . . . . . . . . . . . . . . 67

Other Hormones. . . . . . . . . . . . . . . . . . . . . . 67

Immunoproteins . . . . . . . . . . . . . . . . . . . . . . . 68

Antigens (Vaccines). . . . . . . . . . . . . . . . . . . . 68

Interferon . . . . . . . . . .. . . $ . . . . . . . . . . . . 70

Lymphokines and Cytokines . . . . . . . . . . . . . 71

Antibodies . . . . . . . . . . . . . . . . . . . . . . . . . . 71

Enzymes and Other Proteins . . . . . . . . . . . . . . 72

Enzymes . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72

Other Proteins . . . . . . . . . . . . . . . . . . . . . . . 74

Antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75

Nonprotein Pharmaceuticals . . . . . . . . . . . . . . 76

Impacts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77

Technical Notes . . . . . . . . . . . . . . . . . . . . . . . . . 80

Tables

Table No. Page

3. Large Human Polypeptides Potentially

Attractive for Biosynthesis . . . . . . . . . . . . . . . 61

Table No. Page

4. Naturally Occurring Small Peptides of

Potential Medical Interest . . . . . . . . . . . . . . . . 64

5, Summary of Potential Methods for

Interferon Production. . . . . . . . . . . . . . . . . . . 70

6. Immunoassays . . . . . . . . . . . . . . . . . . . . . . . . 73

7. Diseases Amenable to Drugs Produced by

Genetic Engineering in the Pharmaceutical

Industry . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78

8. Major Diseases for Which Vaccines Need To

Be Developed . . . . . . . . . . . . . . . . . . . . . . . . . 78

Figures

.Figure No. Page

20. The Development of a High PenicillinProducing Strain via Genetic Manipulation . . 60

21. The Product Development Process for

Genetically Engineered Pharmaceuticals . . . . 63

22. The Amino Acid Sequence of Proinsulin . . . . 65

23. Recombinant DNA Strategy for Making

Foot-and-Mouth Disease Vaccine . . . . . . . . . . 69
Chapter 4

The Pharmaceutical Industry

,

Background

The domestic sales of prescription drugs by

U.S. pharmaceutical companies exceeded $7.5

billion in 1979. Of these, approximately 20 percent were products for which fermentation

processes played a significant role. They included anti-infective agents, vitamins, and biological, such as vaccines and hormones. Genetics is expected to be particularly useful in the

production of these pharmaceuticals and biological) which can only be obtained by extraction from human or animal tissues and fluids.

Although the pharmaceutical industry was

the last to adopt traditional fermentation technologies, it has been the first industry to make

widespread use of such advanced genetic technologies as recombinant DNA (rDNA) and cell

fusion. Two major factors triggered the use of

genetics in the pharmaceutical industry:

●

●

The biological sources of many pharmacologically active products are micro-organisms, which are readily amenable to genetic engineering.

The major advances in molecular genetic

engineering have been made under an institutional structure that allocates funds

largely to biomedical research. Hence, the

Federal support system has tended to foster studies that have as their ostensible goal

the improvement of health.

Two factors, however, have tended to discourage the application of genetics in the chemcal and food industries. In the former, economc considerations have not allowed biological

production systems to be competitive with the

existing forms of chemical conversion, with

rare exceptions. And in the latter, social and institutional considerations have not favored the

development of foods to which genetic engineering might make a contribution.

Past uses of genetics

Genetic manipulation of biological systems

for the production of pharmaceuticals has two

general goals:

1. to increase the level or efficiency of the

production of pharmaceuticals with proven or potential value; and

2. to produce totally new pharmaceuticals

and compounds not found in nature.

The first goal has had the strongest influence

on the industry. It has been almost axiomatic

that if a naturally occurring organism can produce a pharmacologically valuable substance,

genetic manipulation can increase the output.

The

●

●

●

following are three classic examples.

The genetic improvement of penicillin production is an example of the elaborate longterm efforts that can lead to dramatic

increases. The original strains of Penicillium chrysogenum, NRRL-1951) were treated

with chemicals and irradiation through

successive stages, as shown in figure 20,

until the strain E-15.1 was developed. This

strain had a 55-fold improvement in productivity over the fungus in which penicillin was originally recognized–the Fleming

strain.

Chemically induced mutations improved a

strain of Escherichia coli to the point where

it produced over 100 times more L-asparaginase (which is used to fight leukemia) than

the original strain. This increase made the

task of isolating and purifying the pharmaceutical much easier, and resulted in lowering the cost of a course of therapy from

nearly $15,000 to approximately $300.

Genetic manipulation sufficiently improved

the production of the antibiotic, gentami59
60 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Figure 20.—The Development of a High Penicillin=

Producing Strain via Genetic Manipulation

First [

mutant —

strain

Wis. Q-176

Legend

Methods used to create each

subsequent mutant in line.

S: natural selection

UV: ultraviolet radiation

X: X-ray radiation

NM: nitrogen mustard

[methyl bis

(&chlorethyarnine]

SA: sarcolyeine

(2-chloroethyl)

amlnophenylalanine]

An illustration of the extensive use of genetics to increase

the yield of a commercially valuable substance. A variety

of laboratories and methods were responsible for the successful outcome.

cin, so that Schering-Plough, its producer,

did not have to build a scheduled manufacturing plant, thereby saving $50 million.

Most industry analysts agree that, overall,

genetic manipulation has been highly significant

in increasing the availability of many pharmaceuticals or in reducing their production costs.

The second major goal of genetic manipulation, the production of new compounds, has

been achieved to a lesser degree. A recent new

antibiotic, deoxygentamicin, was obtained by

mutation and will soon be clinically tested in

man. Earlier, an important new antibiotic,

amikacin, was produced through classical molecular genetic techniques. And before that, the

well-known antibiotic, tetracycline, which is

normally not found in nature, was produced by

a strain of the bacterium, Streptomyces, after

appropriate genetic changes had been carried

out in that bacterium.

SOURCE: Adapted by Office of Technology Assessment from R. P. Elander in

Genetics of Industrial Microorganisms, O. K. Sebek and A. 1. Laekin

(eds.) (Washington, D.C.: American Society for Microbiology, 1979),

P. 23.
Ch. 4—The Pharmaceutical Industry ● 61

Potential uses of molecular genetic technologies —

Polypeptides—proteins—are the first abundant end products of genes. They include peptide hormones, enzymes, antibodies, and certain vaccines. Producing them is the goal of

most current efforts to harness genetically

directed processes. However, it is just a matter

of time and the evolution of technology before

complex nonproteins like antibiotics can also be

manufactured through rDNA techniques.

Hormones

The most advanced applications of genetics

today, in terms of technological sophistication

and commercial development, are in the field of

hormones, the potent messenger molecules that

help the body coordinate the actions of various

tissues. (See Tech. Note 1, p. 80.) The capacity to

synthesize proteins through genetic engineering has stemmed in large part from attempts to

prepare human peptide hormones (like insulin

and growth hormone). The diseases caused by

their deficiencies are presently treated with extracts made from animal or human glands.

The merits of engineering other peptide hormones depend on understanding their actions

and those of their derivatives and analogs.

Evidence that they might be used to improve

the treatment of diabetes, to promote wound

healing, or to stimulate the regrowth of nerves

will stimulate new scientific investigations.

Other relatively small polypeptides that influence the sensation of pain, appetite suppression,

and cognition and memory enhancement are

also being tested. If they prove useful, they will

unquestionably be evaluated for production via

fermentation.

While certain hormones have already attained a place in pharmacology, their testing

and use has been hindered to some extent by

their scarcity and high cost. Until recently,

animal glands, human-cadaver glands, and

urine were the only sources from which they

could be drawn. Their use is also limited

because polypeptide hormones must be administered by injection. They are digested if

they are taken orally, a process that curtails

their usefulness and causes side-effects.

There are four technologies for producing

polypeptide hormones and polypeptides:

. extraction from human or animal organs,

serum, or urine;

● chemical synthesis;

● production by cells in tissue culture; and

● production by microbial fermentation after

genetic engineering.

One major factor in deciding which technology is best for which hormone is the length of

the hormone’s amino acid chains. (See table 3.)

Modern methods of chemical synthesis have

made the preparation of low-molecular weight

polypeptides a fairly straightforward task, and

chemically synthesized hormones up to at least

32 amino acids (AA) in length—like calcitonin

Table 3.—Large Human Polypeptides Potentially

Attractive for Biosynthesis

Amino acid Molecular

residues weight

Proactin . . . . . . . . . . . . . . . . . . . . .

Placental Iactogen . . . . . . . . . . . . .

● Growth hormone. . . . . . . . . . . . . .

Nerve growth factor . . . . . . . . . . . .

Parathyroid hormone (PTH). . . . . .

Prokm.din . . . . . . . . . . . . . . . . . . . .

Insulin-like growth factors

(IGF-I & IGF-2) . . . . . . . . . . . . . . .

Epidermal growth factor . . . . . . . .

● insulin . . . . . . . . . . . . . . . . . . . . . .

Thymopoietin . . . . . . . . . . . . . . . . .

Gastric inhibitory polypeptide

(GIP) . . . . . . . . . . . . . . . . . . . . . . .

● Corticotropin (ACTH) . . . . . . . . . .

Cholecystokinin (CCK-39) . . . . . . .

Big gastrin (BG).. . . . . . . . . . . . . . .

Active fragment of PTH . . . . . . . . .

Cholecystokinin (CCK-33) . . . . . . .

● Calcitonin . . . . . . . . . . . . . . . . . . .

Endorphins . . . . . . . . . . . . . . . . . . .

● Glucagon . . . . . . . . . . . . . . . . . . . .

Thymosin-a1 . . . . . . . . . . . . . . . . . .

Vasoactive intestinal peptide (VIP)

● Secretin. . . . . . . . . . . . . . . . . . . . .

● Active fragment of ACTH. . . . . . .

Motilin . . . . . . . . . . . . . . . . . . . . . . .

● Currently used in medical practice.

SOURCE: Office of Technology Assessment.

198

192

191

118

84

82

70,67

51

49

43

39

39

34

34

33

32

31

29

28

28

27

24

22

22,005

13,000

9,562 bovine

7,649,7471

6,100

5,734

5,104 porcine

4,567 porcine

4,109 bovine

3,918 porcine

3,421 human

3,435 salmon

3,465

3,483 porcine

3,108

3,326 porcine

2,698
62 . Impacts o/Applied Genetics—Micro-Organisms, Plants, and Animals

–have become competitive with those derived

from current biological sources. Since fragments of peptide hormones often express activities comparable or sometimes superior to the

intact hormone, a significant advantage of

chemical synthesis for research purposes is that

analogs having slight pharmacological differences from natural hormones can be prepared

by incorporating different amino acids into

their structures. In principle however, genetically engineered biosynthetic schemes can be

devised for most desirable peptide hormones

and their analogs, although the practicality of

doing so must be assessed on a case-by-case

basis. Ultimately, the principal factors bearing

on the practicality of the competing alternatives

are:

●

●

●

●

The cost of raw materials. For genetically

engineered biosynthesis, this includes the

cost of the nutrient broth plus some amortization of the cost of developing the synthetic organism. In the case of chemical

synthesis, it includes the cost of the pure

amino acid subunits plus the chemicals

used as activating, protecting, coupling, liberating, and supporting agents in the process.

The different costs of separating the desired product from the cellular debris and

the culture medium in biological production, and from the supporting resin, byproducts, and excess reagents in chemical

synthesis.

The cost of purification and freedom from

toxic contaminants. The process is more

expensive for biologically produced material than for materials produced by conventional chemistry, although hormones from

any source can be contaminated.

Differences in the costs of labor and equipment. Chemical synthesis involves a sequence of similar (but different) operations

during a time period roughly proportional

to the length of the amino acid chain (three

AA per day) in an apparatus large enough

to produce 100 grams (g) to 1 kilogram (kg)

per batch; biological fermentations use vats

—with capacities of several thousand gallons–for a few days, regardless of the

length of the amino acid chain.

. The cost and suitability of comparable

materials gathered from organs or fluids

obtained from animals or people.

In the past decade, some simpler hormones

have been chemically synthesized and a few are

being marketed. However, synthesizing glycoproteins—proteins bound to carbohydrates—is

still beyond the capabilities of chemists. Data

obtained from companies directly involved in

the production of peptides by chemical synthesis indicate that the cost of chemically preparing

polypeptides of up to 50 AA in length is extremely sensitive to volume (see Tech. Note 2, p.

80,); although the costs are high, the production

of large quantities by chemical synthesis offers

a competitive production method.

Nevertheless, rDNA production, also known

as molecular cloning, has already been used to

produce low-molecular weight polypeptides. In

1977, researchers at Genentech, Inc., a small

biotechnology company in California, inserted a

totally synthetic DNA sequence into an E. coli

plasmid and demonstrated that it led to the production of the 14 AA polypeptide sequence corresponding to somatostatin, a hormone found in

the brain. The knowledge of somatostatin’s

amino acid sequence made the experiment possible, and the existence of sensitive assays allowed the hormone’s expression to be detected.

Although the primary motive for using this particular hormone for the first demonstration was

simply to show that it could be done, Genentech

has announced that it plans to market its

genetically engineered molecule for research

purposes. (See figure 21.)

Somatostatin is one of about 20 recognized

small human polypeptides that can be made

without difficulty by chemical synthesis. (See

table 4.) Unless a sizable market is found for one

of them, it is unlikely that fermentation meth

ods will be developed in the foreseeable future

Some small peptides that may justify the devel

opment of a biosynthetic process of production

are:

● The seven AA sequence known as MSH

ACTH 4-10, which is reputed to influence

memory, concentration, and other psyche

logical-behavioral effects: should such
Ch. 4—The Pharmaceutical Industry ● 63

Figure 21.—The Product Development Process for Genetically Engineered Pharmaceuticals

Micro-organisms such as E. coli

The development process begins by obtaining DNA either through organic synthesis (1) or derived from biological sources such as tissues

(2). The DNA obtained from one or both sources is tailored to form the basic “gene” (3) which contains the genetic information to “code” for a

desired product, such as human interferon or human insulin. Control signals(4) containing plasmids (6) are isolated from micro-organisms such

as E. coli; cut open (7) and spliced back (8) together with genes and control signals to form “recombinant DNA” molecules. These molecules are

then introduced into a host cell (9).

Each plasm id is copied many times in a cell (10). Each cell then translates the information contained in these plasmids into the desired product, a process called “expression” (11). Cells divide (12) and pass on to their offspring the same genetic information contained in the parent

cell.

Fermentation of large populations of genetically engineered micro-organisms is first done in shaker flasks (13), and then in small fermenters

(14) to determine growth conditions, and eventually in larger fermentation tanks (15). Cellular extract obtained from the fermentation process is

then separated, purified (16), and packaged (17) for health care applications.

Health care products are first tested in animal studies (18) to demonstrate a product’s pharmacological activity and safety, In the United

States, an investigational new drug application (19) is submitted to begin human clinical trials to establish safety and efficacy. Following

clinical testing (20), a new drug application (N DA) (21) is filed with the Food and Drug Administration (FDA). When the NDA has been reviewed

and approved by the FDA the product maybe marketed in the United States (22).

SOURCE: Genentech, Inc.
64 . Impacts of Applied Genetics—Micro-0rganisms, Plants, and Animals

Table 4.-Naturally Occurring Small Peptides of

Potential Medical interest

Number of Molecular

amino acids weight

Dynorphin . . . . . . . . . . . . . . . . . . . . 17

Little gastrin (LG) . . . . . . . . . . . . . . 17

Somatostatin. . . . . . . . . . . . . . . . . . 14

Bombesin . . . . . . . . . . . . . . . . . . . . 14

Melanocyte stimulating hormone. 13

Active dynorphin fragment . . . . . . 13

Neurotensin . . . . . . . . . . . . . . . . . . 13

Mini-gastrin (G13) . . . . . . . . . . . . . . 13

Substance? . . . . . . . . . . . . . . . . . . . 11

Luteinizing hormone-releasing

hormone(LNRH). . . . . . . . . . . . . 10

Active fragment of CCK. . . . . . . . . 10

Angiotensin l . . . . . . . . . . . . . . . . . . 10

Caerulein . . . . . . . . . . . . . . . . . . . . . 10

Bradykinin . . . . . . . . . . . . . . . . . . . . 9

● Vasopressin(ADH) . . . . . . . . . . . . 9

● Oxytocin . . . . . . . . . . . . . . . . . . . . 9

Facteur thymique serique(FTH).. 9

Substance P(4-11)octapeptide. . . 8

Angiotensin lI . . . . . . . . . . . . . . . . . 8

Angiotensin Ill. . . . . . . . . . . . . . . . . 7

MSH/ACTH4-10 . . . . . . . . . . . . . . . 7

Enkephalins. . . . . . . . . . . . . . . . . . . 5

Active fragment of thymopoietin

(TP5). . . . . . . . . . . . . . . . . . . . . . . 5

● Thyrotropin releasing hormone

(TRY) . . . . . . . . . . . . . . . . . . . . . . 3

2,178

1,639

1,620

1,655

l,347bovine

1,297

l,252porcine

1,060

1,007

1,046

931

575

362

.Currently used in medical practice.

SOURCE: Office of Technology Assessment

agents prove of value in wider testing, they

have an enormous potential for use.

Both cholecystokinin (33 AA) and bombesin

(10 AA), which have been shown to suppress appetite, presumably as a satiety

signal from stomach to brain: there is a

large market for antiobesity agents–approximately $85 million per year at the

manufacturer’s level.

Several hormones, such as somatostatin,

which are released by nerves in the hypothalamus of the brain to stimulate or inhibit release of hormones by the pituitary

gland: hormones produced by these glands

are crucial in human fertility; analogs of

some are being investigated as possible

contraceptives.

Calcitonin (32 AA), which is currently the

largest polypeptide produced by chemical

synthesis for commercial pharmaceutical

use: it is useful for pathologic bone disorders, such as Paget’s disease, that affect

up to 3 percent of the population over 40

years of age, in WesternEurope.

. Adrenocorticotropic hormone (ACTH) (39

AA), which promotes and maintains the

normal growth and development of the

adrenalglands and stimulates the secretion

of other hormones: in the United States,

ACTH is used primarily as a diagnostic

agent for adrenal insufficiency, but in

principle, ACTH might be used for at least

one-third of the medical indications—like

rheumatic disorders, allergic states, and

eye inflammation—for which about 5 million Americans annually receive corticosteroids.

Within the last 5 years, other small polypeptides have been identified in many tissues and

have been linked to a variety of activities. Some

certainly bind to the same receptor sites as the

pain-relieving opiates related to the morphine

family. These peptides are called endogenous

opiates: the smaller (5 AA) peptides are called

enkephalins and the larger (3 AA), endorphins.

Certain enkephalins produce brief analgesia

when injected directly into the brains of mice.

Synthetic analogs that are less susceptible to enzymatic inactivation produce longer analgesia

even if they are injected intravenously, as does

the larger endorphin molecule. Very recently,

a 17 AA polypeptide, dynorphin, was reported

to be the most potent pain killer yet found—it is

1,200 times more powerful than morphine.

The preparation of new analgesic agents appears a likely outcome of the new research, but

problems similar to those associated with classical opiates must be overcome. Consequently,

unnatural analogs—including some made with

amino acids not found in micro-organisms—

might prove more useful. The value of microbial biosynthesis for these substances is questionable at this time. However, the importance

of genetic technologies in clarifying the

underlying mechanisms should not be underestimated.

Higher molecular weight polypeptides cannot

be made practically by chemical synthesis, and

must be extracted from human or animal tissues or produced in cells growing in culture.
Ch. 4—The Pharmaceutical Industry ● 65

Now they can also be manufactured by fermentation using genetically designed bacteria, as

has been demonstrated by the production of insulin and human growth hormone.

INSULIN

Insulin, is composed of two chains—A and B—

of amino acids. It is initially produced as a

single, long chain called pre-proinsulin, which is

cut into a shorter chain, proinsulin. Proinsulin,

in turn, is cut into the A and B chains when a

piece is cleaved from the middle. (See figure 22.)

Work on the genetic engineering of insulin has

proceeded quickly. A year after one group reported that the insulin gene had been incorporated into E. coli without expression, a second

group managed to grow colonies of E. coli that

actually excreted rat proinsulin. Then, within a

couple of months, workers at Genentech, in collaboration with a group at City of Hope Medical

Center, announced the separate synthesis of the

A (21 AA) and B (30 AA) chains of human insulin.

The synthesis of the DNA sequences depended

on advances in organic chemistry as well as in

genetics. Six months were required simply to

synthesize the necessary building blocks.

Figure 22.—The Amino Acid Sequence of Proinsulin

Connecting peptide

N H

+

3

B chain

Proinsulin is composed of 84 amino acid residues. When the connecting peptide is removed,

the retaining A and B chains form the insulin molecule. The A chain contains 21 amino acids;

the B chain contains 30 amino acids.

SOURCE: Office of Technology Assessment.
66 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

A comparison with the traditional source of

animal insulin is interesting. If 0.5 milligram

(mg) of pure insulin can be obtained from a liter

of fermentation brew, 2,000 liters (1) (roughly

500 gal) would yield 1 g of purified insulin—the

amount produced by about 16 lb of animal pancreas. If, on the other hand, the efficiency of

production could be increased to that achieved

for asparaginase (which is produced commercially by the same organism, E. coli), 2,000

would yield 100 g of purified insulin—the

amount extracted from 1,600 lb of pancreas.

(The average diabetic uses the equivalent of

about 2 mg of animal insulin per day.)

The extent of the actual demand for insulin is

a controversial issue. Eli Lilly & Co. estimates

that there are 60 million diabetics in the world

(35 million in underdeveloped countries, where

few are diagnosed or treated). Of the 25 million

in the developed countries, perhaps 15 million

have been diagnosed; according to Lilly’s estimate, 5 million are treated with insulin. Only

one-fourth of those diabetics treated with insulin live in the United States, but they use 40 to

50 percent of the insulin consumed in the

world. A number of studies indicate that while

the emphasis on diet (alone) and oral antidiabetic drugs varies, approximately 40 percent of

American patients in large diabetes clinics or

practices take insulin injections. In the United

States, diabetes ranks as the fifth most common

cause of death and second most common cause

of blindness. Roughly 2 million persons require

daily injections of insulin.

Today, at least, there is no real shortage of

glands from slaughter houses for the production of animal (principally bovine and porcine)

insulin. A study conducted by the National Diabetes Advisory Board (NDAB) concluded that a

maximum demand and a minimum supply

would lead to shortages in the 1990’s. Eli Lilly’s

projection, presented in that report, also anticipates these shortages. But, Novo Industri, a major world supplier of insulin, told the NDAB that

it estimates that the 1976 free-world consumption of insulin of 51 X 109 units constituted only

23 percent of the potential supply, and the

87X 109 units projected for 1996 would only

equal 40 percent of the supply, assuming that

the animal population stays constant.

For insulin, therefore, the limitation on bringing the fruits of genetic engineering to the

marketplace is not technological but institutional. The drug must first be approved by the

Food and Drug Administration (FDA) and then

marketed as a product as good as or better than

the insulin extracted by conventional means.

Lilly has stated that it anticipates a 6-month

testing period in humans. Undoubtedly, FDA

will examine the evidence presented in the investigational new drug application (INDA) with

special care. Its review will establish criteria

that may influence the review of subsequent applications in at least the following requirements:

evidence that the amino acid sequence of

the material is identical to that of the normal human hormone;

freedom from bacterial endotoxins that

may cause fever at extremely low concentrations—an inherent hazard associated

with any process using E. coli; and

freedom from byproducts, including substances of very similar structure that may

give rise to rare acute or chronic reactions

of the immune system.

Furthermore, as development continues, FDA

might require strict assurances that the molecules produced from batch to batch are not subject to subtle variations resulting from their

genetic origin.

If the insulin obtained from rDNA techniques

manages to pass FDA requirements, it must

overcome a second obstacle—competition in the

marketplace. The clinical rationale for using

human rather than animal insulin rests on the

differences in structure among insulins produced by different species. Human and porcine

insulins for example, differ in a single amino

acid, while human and cattle insulins differ

with respect to three. As far as is known, these

variations do not impair the effectiveness of the

insulin, but no one has ever been in a position to

conduct a significant test of the use of human

insulin in a diabetic population. Many consequences of the disease, such as retinopathy (retinal disease) and nephropathy (kidney disease),

are not prevented by routine injection of animal

insulin. Patients also occasionally respond adversely or produce antibodies to animal insulin,

with subsequent allergic or resistant reaction.
Ch. 4—The Pharmaceutical Industry . 67

It remains to be seen how many patients will

be better off with human insulin. The proof that

it improves therapy will take years. Progress on

the etiology of the disease—especially in identifying it in those at risk or in improving the

dosage form and administration of insulin–may

have far more significant effects than new developments in insulin production. Nevertheless,

as long as private enterprise sees fit to invest in

such developments, and as long as the cost of

treating diabetics who respond properly to animal insulin is not increased, biological production of human insulin may become a kind of insurance for diabetics within the next few

decades.

GROWTH HORMONE

The second polypeptide hormone currently a

candidate for FDA approval is growth hormone

(GH). It is one of a family of closely related, relatively large pituitary peptide hormones—single-chain polypeptides 191- to 198-AA in length.

It is best known for the growth it induces in

many soft tissues, cartilage, and bone, and it is a

requirement for postnatal growth in man.

The growth of an organism is a highly complex process that depends on the correct balance of many variables: The action of GH in the

body for example, depends on the presence of

insulin, whose secretion is stimulated by GH.

Under some circumstances, one or more intermediary polypeptides produced under the influence of GH by the liver (and possibly the

kidneys) may actually be the proximate causes

of some of the effects attributed to GH. In any

case, the biological significance of GH is most

clearly illustrated by the growth retardation

that characterizes its absence before puberty,

and by the benefits of replacement therapy.

In the United States, most of the demand for

human growth hormone (hGH) is met by the National Pituitary Agency, which was created in

the early 1960’s by the College of Pathologists

and the National Institute of Arthritis, Metabolism, and Digestive Diseases (NIAMDD) to collect pituitary glands from coroners and private

donors. Under the programs of the NIAMDD,

hGH is provided without charge to treat children with hypopituitarism, or dwarfism (about

1,600 patients, each of whom receives therapy

for several years), and for research.

While the National Pituitary Agency feels that

it can satisfy the current demand for hGH (see

Tech. Note 3, p. 80.), it welcomes the promise of

additional hGH at relatively low cost to satisfy

areas of research that are handicapped more by

a scarcity of funds than by a scarcity of the hormone. However, if hGH is shown to be therapeutically valuable in these areas, widespread

use could severely strain the present supply. At

present, the potential seems greatest for patients with:

●

●

●

●

●

senile osteoporosis (bone decalcification);

other nonpituitary growth deficiencies such

as Turner’s syndrome (1 in 3,000 live

female births);

intrauterine growth retardation;

bleeding ulcers that cannot be controlled

by other means; and

burn, wound, and bone-fracture healing

Two groups have already announced the

preparation of micro-organisms with the capacity for synthesizing GH. (See Tech. Note 4, p. 80.)

In December 1979, one of these groups—Genentech —requested and received permission from

the National Institutes of Health (NIH), on the

recommendation of the Recombinant DNA Advisory Committee (RAC), to scale-up its process.

Its formation of a joint-venture with Kabi Gen

AB is typical of the kind of alliance that develops

as a result of the different expertise of groups in

the multidisciplinary biomedical field. Kabi has

been granted a New Drug Application (NDA)

under which to market pituitary GH imported

from abroad.

OTHER HORMONES

Additional polypeptide hormones targeted

for molecular cloning (rDNA production) include:

●

●

Parathyroid hormone (84 AA), which may

be useful alone or in combination with calcitonin for bone disorders such as osteoporosis.

Nerve growth factor (118 AA), which influences the development, maintenance, and
68 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

repair of nerve cells and thus could be significant for nerve restoration in surgery.

● Erythropoietin, a glycopeptide that is largely responsible for the regulation of blood

cell development. Its therapeutic applications may range from hemorrhages and

burns to anemias and other hematologic

conditions. (See Tech. Note 5, p. 80.)

Immunoproteins

Immunoproteins include all the proteins that

are part of the immune system—antigens, interferon, cytokines, and antibodies. Since polypeptides, the primary products of every molecular cloning scheme, are at the heart of immunology, developments made possible by recent

breakthroughs will presumably affect the entire

field. There is little doubt that applied genetics

will play a critical role in developing a pharmacology for controlling immunologic functions,

since it provides the only apparent means of

synthesizing many of the agents that will comprise immunopharmacology.

ANTIGENS (VACCINES)

One early dramatic benefit should be in the

area of vaccination, where genetic technologies

may lead to the production of harmless substances capable of eliciting specific defenses

against various stubborn infectious diseases.

Vaccination provides effective immunity by

introducing relatively harmless antigens into

the immune system thereby allowing the body

to establish, in advance, adequate levels of antibody and a primed population of cells that can

grow when the antigen reappears in its virulent

form. Obviously, however, the vaccination itself

should not be dangerous. As a result, several

methods have been developed over the past two

centuries to modify the virulence of micro-organisms used in vaccines without destroying their

ability to trigger the production of antibodies.

(See Tech. Note 6, p. 80.)

Novel pure vaccines based on antigens synthesized by rDNA have been proposed to fight

communicable diseases like malaria, which have

resisted classical preventive efforts. Pure vaccines have always been scarce; if they were

available, they might reduce the adverse effects

of conventional vaccines and change the methods and the dosages in which vaccines are

administered.

Some vaccines are directed against toxic proteins (like the diphtheria toxin produced by

some organisms), preparing the body to neutralize them. Molecular cloning might make it possible to produce inactivated toxins, or better

nonvirulent fragments of toxins, by means of

micro-organisms that are incapable of serving

as disease-causing organisms.

Immunity conferred by live vaccines invariably exceeds that conferred by nonliving antigenic material–possibly because a living microorganism creates more antigen over a longer

period of time, providing continuous “booster

shots.” Engineered micro-organisms might become productive sources of high-potency antigen, offering far larger, more sustained, doses

of vaccine without the side-effects from the contaminants found in those vaccines that consist

of killed micro-organisms.

However, it is clear that formidable Federal

regulatory requirements would have to be met

before permission is granted for a novel living

organism to be injected into human subjects.

Because of problems encountered with live vaccines, the most likely application will lie in the

area of killed vaccines (often using only parts of

micro-organisms).

It is impossible in the scope of this report to

discuss the pros, cons, and consequences of developing a vaccine for each viral disease. However, the most commercially important are the

influenza vaccines, with an average of 20.8 million doses given per year from 1973 to 1975–a

smaller number than the 25.0 million doses per

year of polio vaccine, but more profitable.

Influenza is caused by a virus that has remained uncontrolled largely because of the frequency with which it can mutate and change its

antigenic structures. It has been suggested that

antigenic protein genes for influenza could be

kept in a “gene bank” and used when needed. In

addition, the genetic code for several antigens

could be introduced into an organism such as E.
Ch. 4—The Pharmaceutical Industry ● 69

coli, so that a vaccine with several antigens

might be produced in one fermentation. 1

Two more viral diseases deserve at least brief

comment. Approximately 800 million doses of

foot-and-mouth disease virus (FMDV) vaccine

are annually used worldwide, making it the

largest volume vaccine produced. This vaccine

must be given frequently to livestock in areas

where the disease is endemic, which includes

most of the world outside of North America.

The present methods of producing the vaccine

require that enormous quantities of hazardous

virus be contained. Many outbreaks are attributed to incompletely inactivated vaccine or to

the escape of the virus from factories. (See

figure 23.)

Molecular cloning of the antigen could produce a stable vaccine at considerably less expense, without the risk of the virus escaping. On

the basis of that potential, RAC has approved a

joint program between the U.S. Department of

Agriculture (USDA) and Genentech to clone

pieces of the FMDV genome to produce pure antigen. The RAC decision marked the first exception to the NIH prohibition against cloning DNA

that is derived from a virulent pathogen.2 FMDV.

vaccine made by molecular cloning will probably be distributed commercially by 1985, although not in the United States. It will be the

first vaccine to achieve that status, and illustrates the potential veterinary uses of genetic

technologies.

Hepatitis has also received significant attention. Vaccines against viral hepatitis, which affects some 300,000 Americans each year, may

be produced by molecular cloning. This disease

is second only to tuberculosis as a cause of

death among reportable infectious diseases. R is

extremely difficult to cultivate the causative

agents. Hepatitis A has a good chance of being

the first human viral disease for which the initial preparation of experimental vaccine will involve molecular cloning. A vaccine against hepatitis B, made from the blood of chronic carriers,

I For other aspects of vaccine production see: Office of Technology Assessment, U.S. Congress, working Papers, The impacts of

Generics, vol. 2. (Springfield, Va.: National Technical Information

Service, 1981).

‘Ibid.

Figure 23.-Recombinant DNA Strategy for Making

Foot-and-Mouth Disease Vaccine

I

Growing E. co/i bacteria may produce VP3 for use as vaccine

for foot-and-mouth disease. No virus or infectious RNA is

produced by the harmless bacteria strain.

● VP3 is the protein from the shell of the virus, which can act

as a vaccine for immunizing livestock against foot-andmouth disease. The idea outlined above is to make this VP3

protein without making any virus or infectious RNA.

SOURCE: Office of Technology Assessment.

is in the testing stage, but cloning is being investigated as a better source of an appropriate

antigen. The causative agent for a third form of

hepatitis has not even been identified. Since at

least 16 million U.S. citizens are estimated to be

at high risk of contracting hepatitis, there is

keen interest in the development of vaccines

among academic and industrial researchers.3

31bid.

76-565 0 - 81 - 6
70 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

More hypothetically, molecular cloning may

lead to three other uses of antigens as well: vaccination against parasites, such as malaria and

hookworm (see Tech. Note 7, p. 80.); immunization in connection with cancer treatment; and

counteracting abnormal antibodies, which are

made against normal tissues in the so-called

“autoimmune diseases, ” such as multiple sclerosis. (See Tech. Note 8, p. 81.)

INTERFERON

Interferon are glycoproteins normally made

by a variety of cells in response to viral infection. All interferon (see Tech. Note 9, p. 81) can

induce an antiviral state in susceptible cells. In

addition, interferon has been found to have at

least 15 other biochemical effects, most of

which involve other elements of the immune

system.

promising preliminary studies have supported the use of interferon in the treatment of

such viral diseases as rabies, hepatitis, varicellazoster (shingles), and various herpes infections.

To date, the effect of interferon has been far

more impressive as a prophylactic than as a

therapeutic agent. The interferon produced by

Genentech, for example, has been shown to protect squirrel monkeys from infection by the lethal myocarditis virus. Once interferon is available in quantity, large-scale tests on human populations can be conducted to confirm its efficacy in man.

Several production techniques are being explored. (See Tech. Note 10, p. 81.) Extraction of

interferon from leukocytes (white blood cells),

the current method of choice, may have to compete with tissue culture production as well as

rDNA. (See table 5.)

Recombinant DNA is widely regarded as the

key to mass production of interferon, and

important initial successes have already been

achieved. Each of the four major biotechnology

companies is working on improved production

methods, and all have reported some success.

An enormous amount remains to be learned

about the interferon system. It now appears

that the interferon are simply one of many

families of molecules involved in physiological

regulation of response to disease. Only now

have molecular biology and genetics made their

study—and perhaps their use—possible.

Table 5.—Summary of Potential Methods for Interferon Production

Types of Potential

interferon for Present projected Potential for

Means of Production produced scale-up ($/106 units) Problems improvement

“Buffy coat” leukocytes leukocyte, 95% No 50 –

fibroblast, 5%

Lymphoblastoid cells leukocyte, 80% Yes — =25

fibroblast, 20%

Fibroblasts fibroblast Yes 43-200 = 1-10

Recombinant DNA leukocyte or

fibroblast

Yes — z 1-10

—lack of scale up

—pathogen contamination

—poor yields

—cells derived from tumor

—cell culture

—economic competition

with recombinant DNA

—does not produce

interferon

—in vitro drug stability

—poor yields

—minimal

—improved yields

—expression of

fibroblast

interferon

—improved yields

—improved cellculture

technology

—expression of

leukocyte-type

interferon

—improved yields

—modified

interferon

—drug approval

—possible economic

competition with fibroblast

cell production

SOURCE: Office of Technology Assessment.
Ch. 4—The Pharmaceutical Industry . 71

The interferon are presently receiving attention largely because studies in Sweden and the

United States stimulated the appropriation of

$5.4 million by the American Cancer Society

(ACS) for expanded clinical trials in the treatment of cancer. That commitment by the nonprofit ACS–the greatest by far in its history–

was followed by a boost in NIH funding for interferon research from $7.7 million to $19.9

million for fiscal year 1980. Much of the cost of

interferon research is allotted to procuring the

glycopeptide. Initially, the ACS bought 40 billion

units of leukocyte interferon from the Finnish

Red Cross for $50 per million units. In March

1980, Warner-Lambert was awarded a contract

to supply the National Cancer Institute (NCI)

with 50 billion units of leukocyte interferon

within the next 2 years at an average price of

$18 per million units. NCI is also planning to

purchase 50 billion units each of fibroblast and

lymphoblastoid interferon.

The bulk of the NIH funding is included in

NCI’s new Biological Response Modifier (BRM)

program—interferon accounts for $13.9 million

of the $34.1 million allocated for BRM work in

fiscal year 1980. (NCI expenditures on interferon in 1979 were $2.6 million, 19 percent of

the amount budgeted for 1980.) Other important elements of that BRM program concern

immunoproteins known as lymphokines and

thymic hormones, for which molecular genetics

has major implications. The program is aimed at

identifying and testing molecules that control

the activities of different cell types.

LYMPHOKINES AND CYTOKINES

Lymphokines and cytokines are regulatory

molecules that have begun to emerge from the

obscure fringes of immunology in the past 10

years. (Interferon is generally considered a lymphokine that has been characterized sufficiently

to deserve independent status.)

Lymphokines are biologically active soluble

factors produced by white blood cells. Studied

in depth only within the last 15 years, they are

being implicated at virtually every stage in the

complex series of events that make up the immune response. They now include about 100

different compounds. Cytokines, which have effects similar to lymphokines, include several

compounds associated with the thymus gland,

referred to as thymic hormones.4

In 1979, the BRM subcommittee concluded

that several of these agents probably have great

potential for cancer treatment. Nevertheless,

adequate quantities for laboratory and clinical

testing of many of them will probably not be

available until the problems of producing glycoproteins by molecular cloning are overcome. No

system is currently available for the industrial

production of glycoproteins, although yeasts

may prove to be the most useful micro-organisms.

ANTIBODIES

Antibodies are the best known and most exploited protein components of the immune system. Until recently, all antibodies were obtained

from the blood of humans or animals; and they

were often impure. Within the past 5 years,

however, it has become possible to produce antibodies from cells in culture, and to achieve

levels of purity previously unattainable. As with

previous advances in antibody technology, researchers are examining ways to put this new

level of purity to use. There have been hundreds, if not thousands, of examples of new

diagnostic and research methods, new methods

of purification, and new therapies published

within the first 3 years that the technique has

been available. (See Tech, Note 11, p. 81.)

This high level of purity was attained by the

development of monoclinal antibodies. These

antibodies that recognize only one kind of antigen were the unanticipated fruit of fundamental immunological research conducted by Drs.

Caesar Milstein and Georges Kohler at the Medical Research Council in England in 1975. They

fused two types of cells–myeloma and plasmaspleen cells—to form hybridomas that produce

the monoclinal antibodies. (See Tech. Note 12,

p. 81.) Not only are the antibodies specific, but

because the hybridomas can be grown in mass

culture, a virtually limitless supply is available.

The most immediate medical application for

monoclinal antibodies lies in diagnostic testing.

4

For 40 of the best characterized cytokines, see footnote 1, p.

69.
72 . Impacts of Applied Genetics—Micro-Organ/SmS, Plants, and Animals

Over the past 20 years, large segments of the

diagnostic and clinical laboratory industries

have sprung up to detect and quantify particular substances in specimens. Because monoclonal antibodies are so specific, hybridomas seem

certain to replace animals as the source of antibodies for virtually all diagnosis and monitoring. Their use will not only improve the accuracy of tests and decrease development costs,

but should result in a more uniform product.

Today, such assays are used to:

determine hormone levels in order to

assess the proper functioning of an endocrine gland or the inappropriate production of a hormone by a tumor;

detect certain proteins, the presence of

which has been found to correlate with a

tumor or with a specific prenatal condition;

detect the presence of illicit drugs in a person’s blood, or monitor the blood or tissue

level of a drug to ensure that the dosage

achieves a therapeutic level without exceeding the limits that could cause toxic effects; and

identify microbial pathogens.

The extent of the use of antibodies and the

biochemical properties that they can identify is

suggested by table 6. No one assay constitutes a

major market, and short product lifetime has

been characteristic of this business.

Other applications of monoclinal antibodies

include:

the improvement of the acceptance of kidney (and other organ) transplants by injection of the recipient with antibodies against

certain antigens;

passive immunization against an antigen involved in reproduction, as a reversible immunological approach to contraception.

localizing tumors with tumor-specific antibodies (see Tech. Note 13, p. 81); and

targeting cancer cells with antibodies that

have anticancer chemicals attached to

them.

Enzymes and other proteins

ENZYMES

Enzymes are involved in virtually every biological process and are well-understood. Nevertheless, despite their potency, versatility, and

diversity, they play a small role in the practice

of medicine today. Therapeutic enzymes accounted for American sales of about $70 million

(wholesale) in 1978, but one-half of those sales

involved the blood-plasma-derived coagulation

factors used to treat hemophilia. Although the

figure is difficult to estimate, the total number

of patients receiving any type of enzyme therapy in 1980 probably does not exceed 50,000.

Enzymes cannot be synthesized by conventional chemistry. Almost all those presently

employed in medicine are extracted from

human blood, urine, or organs, or are produced

by micro-organisms. Already the possibility of

using rDNA clones as the source of enzymes—

primarily to reduce the cost of production—is

being explored.

However, problems associated with the use of

nonhuman enzymes (such as immune and febrile responses) and the scarcity of human enzymes, have hindered research, development,

and clinical exploitation of enzymes for therapeutic purposes. Today, the experimental genetic technologies of rDNA and somatic cell fusion and culture open the only conceivable

routes to relatively inexpensive production of

compatible human enzymes.

The genetic engineering of enzymes is probably the best example of a dilemma that hampers the exploitation of rDNA: Without a clinical

need large enough to justify the investment,

there is no incentive to produce a product; yet

without adequate supplies, the therapeutic possibilities cannot be investigated. The substances

that break this cycle will probably be those that

are already produced in quantity from natural

tissue.

The only enzymes administered today are

given to hemophiliacs–and they are actually
Ch. 4—The Pharmaceutical Industry ● 73

Table 6-Immunoassays .

Analgesics and narcotics

Anileridine

Antipyrine

Codeine

Etorphine

Fentanyl

Meperidine

Methadone

Morphine

Pentazocine

Antibiotics

Amikacin

Chloramphenicol

Clindamycin

Gentamicin

Isoniazid

Penicillin

Sisomycin

Tobramycin

Anticonvulsants

Clonazepam

Phenytoin

Primidone

Anti-inflammatory agents

Colchicine

Indomethacin

Phenyibutazone

Antineoplastic agents

Adriamycin

Bleomycin

Daunomycin

Methotrexate

Bronchodilators

Theophylline

Cardiovascular drugs

Cardiac glycosides

Acetylstrophanthidin

Cedilanid

Deslanoside

Digitoxin

Digoxin

Gitoxin

Hallucinogenic drugs

Mescaline

Tetrahydrocannabi nol

Hypoglycemic agents

Butylbiguanide

Glibenclamid

Insecticides

Aldrin

DDT

Dieldrin

Malathion

Narcotic antagonists

Cyclazocine

Naloxone

Peptide hormones

Angiotensin

Anterior pituitary

Bradykinin

Gastric

Hypothalamic

Intestinal

Pancreatic

Parathyroid

Posterior pituitary

Thyroid (calcitonin)

Plant hormones

lndole-3-acetic acid

Gibberelilic acid

Polyamides

Spermine

Prostaglandins

Sedatives and

tranquilizers

Barbiturates

Barbital

Pentobarbital

Phenobarbital

Chlordiazepoxide

Chlorpromazine

Desmethylimipramine

Diazepam and

N-desmethyldiazepam

Methyl digoxin

Ouabain

Proscillaridin

Dihydroergotamine

Propranolol

Quinidine

CNS stimulants

Amphetamine

Benzoyl ecgonine

(cocaine metabolize)

Methamphetamine

Pimozide

Diuretics

Bumetanide

Hallucinogenic drugs

Bile acid conjugates

Cholylglycine

Cholyltaurine

Catecholamines

Epinephrine

Norepinephrine

Tyramine

Fibrinopeptides

Fibrinopeptide A

Fibrinopeptide B

Indolealkylamines

Melatonin

Serotonin

Insect hormones

Ecdysone

Nucleosides and

nucleotides

Cyclic AMP

Cyclic GMP

N

2

-Dimethylguanosine

7-Methylguanosine

Pseudouridine

Thymidine

Glutethimide

Methaqualone

Steroid hormones

Skeletal muscle relaxants

d-Tubocurarine

Synthetic peptides

DDAVP

Saralasin

Synthetic steroids

Anabolic steroids

Trienbolone acetate

Androgens

Fluoxymesterone

Estrogens

Diethylstilbestrol

Ethinylestradiol

Mestranol

Glucocorticoids

Dexamethasone

Methyl prednisolone

Prednisolone

Prednisone

Metyrapone

Progestins

Medroxyprogesterone

acetate

Norethindrone

Norethisterone

Norgestrel

Toxins

Aflatoxin B,

Genistein

Nicotine and metabolizes

Ochratoxin A

Paralytic shellfish poison

Thyroid hormones

Thyroxine

Triodothyronine

Vitamins

Vitamin B12

Vitamin D

SOURCE: ‘“lmmunoassays of Drugs–Comprehensive Immunology,” hrrrnunal Pharmacology, Hadde~ Caffey (cd.) (New York: Plenum Press, 1977), P. 325.

proenzymes, which are converted to active enzymes in the body when needed. The most common agents are called Factor VIII and Factor IX,

which are found in serum albumin and are currently extracted from human blood plasma .

Hemophilia A and Hemophilia B—accounting for

over 90 percent of all major bleeding disorders—are characterized by a deficiency of these

factors. Supplies of the proenzymes will exceed

demand well beyond 1980 if the harvesting and

processing of plasma continues as it has. Nevertheless, the risk of hepatitis associated with the

use of human plasma-derived products is ex -

tremely high. One recent study found chroni c

hepatitis in a significant percentage of asymptomatic patients treated with Factor VIII an d

Factor IX.

The plasma fractionation industry, whic h

produces the proenzymes, is currently face d

with excess capacity, intense competition, high

plasma costs, and tight profit margins. 5 The cost

and availability of any one plasma protein i s

‘For details of the factors governing the industry. see footnote 1 ,

p. 69.
74 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

coupled to the production of the others. Hence,

the industry would still have to orchestrate the

production of the other proteins even if just one

of them, such as Factor VIII, becomes a target

for biological production.

Another enzyme, urokinase, has been targeted for use in removing unwanted blood clots,

which lead to strokes, myocardial infarctions,

and pulmonary emboli. Currently, the drug is

either isolated from urine or produced in tissue

culture. (See Tech. Note 14, p. 81.)

Urokinase is thus far the only commercial

therapeutic product derived from mammalian

cell culture. Nevertheless, some calculations

suggest that production by E. coli fermentation

would have economic advantages. The costs implicit in having to grow cells for 30 days on fetal

calf serum (or its equivalent) or in having to collect and fractionate urine–as reflected in urokinase’s market price ($150/mg at the manufacturer’s level) —should be enough incentive to encourage research into its production. In fact, in

April 1980, Abbott Laboratories disclosed that

E. coli had been induced to produce urokinase

through plasmid-borne DNA.

The availability of urokinase might be guaranteed by the new genetic technologies, but its

use is not. For a variety of reasons, the American medical community has not accepted the

drug as readily as have the European and Japanese communities. Studies to establish the use

of urokinase for deep vein thrombosis, for example, are now being conducted almost exclusively in Europe.6

OTHER PROTEINS

In addition to the proteins and polypeptides

already mentioned, the structural proteins,

such as the collagens (the most abundant proteins in the body), elastins and keratins (the

compounds of extracellular structures like hair

and connective tissue), albumins, globulins, and

a wide variety of others, may also be susceptible

to genetic engineering. Structural proteins are

less likely to be suitable for molecular genetic

manipulations: On the one hand, their size and

eFor additional information about how urokinase came to play a

role in therapy, see footnote 1, p. 69.

complexity exceed the synthetic and analytic

capabilities that will be available in the next few

years; on the other, either their use in medicine

has yet to be established or material derived

from animals appears adequate, as is the case

with collagen, for which uses are emerging.

Plasma, the fluid portion of the blood, contains about 10 percent solids, most of which are

proteins. During World War II, a simple procedure was developed to separate the various

components. It is still used today.

Serum albumin is the smallest of the main

plasma proteins but it constitutes about half of

plasma’s total mass. Its major therapeutic use is

to reverse the effects of shock. p It is a reasonable candidate for molecular cloning, although

its relatively high molecular weight complicates

purification, and its commercial value is relatively low. The market value of normal serum

albumin is approximately $3/g, but the volume

is such that domestic sales exceed $150 million.

Including exports, annual production is in the

range of 100,000 kg.

Normal serum albumin for treating shock is

already regarded as too expensive compared

with alternative treatments, to expand its use

would require a lower price. On the other hand,

the Federal Government—and especially the Department of Defense—might disregard the immediate economic prospects and conclude that

having a source of human serum albumin that

does not depend on payments to blood donors

might be in the national interest. Since many nations import serum albumin, products derived

from molecular cloning could be exported.

Serum albumin is presently the principal

product of blood plasma fractionation, a change

in the way it is manufactured would significantly affect that industry. Because a number of

other products (such as clotting factors) are also

derived from fractionation, a growth in the

need for plasma-derived albumin could have a

significant impact on the availability and the

cost of these byproducts.

7

For a detailed discussion of the costs and benefits of using albumin and the structure of the industry, see footnote 1, p. 69,
Ch. 4—The Pharmaceutical industry . 75

Antibiotics

Antimicrobial agents for the treatment of infectious diseases have been the largest selling

prescription pharmaceuticals in the world for

the past three decades. Most of these agents are

antibiotics—antimicrobials naturally produced

by micro-organisms rather than by chemical

synthesis or by isolation from higher organisms.

However, one major antibiotic, chloramphenicol—originally produced by a micro-organism,

is now synthesized by chemical methods. The

field of antibiotics, in fact, provides most of the

precedent for employing microbial fermentation to produce useful medical substances. The

United States has been prominent in their

development, production, and marketing, with

the result that American companies account for

about half of the roughly $5 billion worth of antimicrobial agents sold worldwide each year.

The American market share has been growing

as new antibiotics are developed and introduced every year.

For 30 years,high-yielding, antibiotic-producing micro-organisms have been identified by

selection from among mutant strains. Initially,

organisms producing new antibiotics are isolated by soil sampling and other broad screening efforts. They are then cultured in the laboratory, and efforts are made to improve their

productivity.

Antibiotics are complex, usually nonprotein,

substances, which are generally the end products of a series of biological steps. While knowledge of molecular details in metabolism has

made some difference, not a single antibiotic

has had its complete biosynthetic pathway elucidated. This is partly because there is no single

gene that can be isolated to produce an antibiotic. However, mutations can be induced within

the original micro-organism so that the level of

production can be increased.

Other methods can also increase production,

and possibly create new antibiotics. Microbial

mating, for example, which leads to natural

recombination, has been widely investigated as

a way of developing vigorous, high-yielding antibiotic producers. However, its use has been

limited by the mating incompatibility of many

industrially important higher fungi, the presence of chromosomal aberrations in micro-organisms improved by mutation, and a number of

other problems. Furthermore, natural recombination is most advantageous when strains of

extremely diverse origins are mated; the proprietary secrets protecting commercial strains

usually prevent the sort of divergent “competitor” strains most likely to produce vigorous

hybrids from being brought together.

The technique of protoplasm or cell fusion

provides a convenient method for establishing a

recombinant system in strains, species, and

genera that lack an efficient natural means for

mating. For example, as many as four strains of

the antibiotic-producing bacterium Streptomyces have been fused together in a single step to

yield recombinant that inherit genes from four

parents. The technique is applicable to nearly

all antibiotic producers. It will help combine the

benefits developed in divergent lines by mutation and selection.

In addition, researchers have compared the

quality of an antibiotic-producing fungus, Cephalosporimn acremonium, produced by mating to

one produced by protoplasm fusion. (See Tech.

Note 15, p. 82.) They concluded that protoplasm

fusion was far superior for that purpose. What

is more, protoplasm fusion can give rise to hundreds of recombinants–including one isolate

that consistently produced the antibiotic cephalosporin C in 40 percent greater yield than the

best producer among its parents–without losing that parent strain’s rare capacity to use inorganic sulfate, rather than expensive methionine, as a source of sulfur. It also acquired the

rapid growth and sporulation characteristics of

its less-productive parent. Thus, desirable attributes from different parents were combined

in an important industrial organism that had

proved resistant to conventional crossing.

Even more significant are the possibilities for

preparation by protoplasm fusion between different species or genera of hybrid strains,

which could have unique biosynthetic capacities. One group is reported to have isolated a

novel antibiotic, clearly not produced by either

parent, in an organism created through fusion

of actinomycete protoplasts, (See Tech. Note 16,
76 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

p. 82.) The value of protoplasm fusion, therefore,

lies in potentially broadening the gene pool.

Protoplasm fusion is genetic recombination on

a large scale, Instead of one or a few genes being transferred across genus and species barriers, entire sets of genes can be moved. Success

is not assured, however; a weakness today is the

inherited instability of the “fused” clones. The

preservation of traits and long-range stability

has yet to be resolved. Furthermore, it seems

that one of the most daunting problems is

screening—determining what to look for and

how to recognize it. (See Tech. Note 17, p. 82.)

Recombinant DNA techniques are also being

examined for their ability to improve strains,

Many potentially useful antibiotics do not reach

their commercial potential because the microorganisms cannot be induced to produce sufficient quantities by traditional methods. The synthesis of certain antibiotics is controlled by

plasmids, and it is believed that some plasmids

may nonspecifically enhance antibiotic production and excretion.

It may also be possible to transfer as a group,

all the genes needed to produce an antibiotic

into a new host. However, increasing the number of copies of critical genes by phage or plasmid transfer has yet to be achieved in antibioticproducing organisms because little is known of

the potential vectors. The genetic systems of

commercial strains will have to be understood

before the newer genetic engineering approaches can be used. Genetic maps have been

published for only 3 of the 24 or more industrially useful bacteria.

Since 2,000 of the 2,400 known antibiotics are

produced by Streptomyces, that is the genus of

greatest interest to the pharmaceutical industry. Probably every company conducting research on Streptomyces is developing vectors,

but little of the industrial work has been revealed to date.

Nonprotein pharmaceuticals

In both sales and quantity, over 80 percent of

the pharmaceuticals produced today are not

made of protein. Instead, they consist of a variety of organic chemical entities. These drugs, except for antibiotics, are either extracted from

some natural plant or animal source or are synthesized chemically.

Some of the raw materials for pharmaceuticals are also obtained from plants; micro-organisms are then used to convert the material to

useful drugs in one or two enzymatic steps.

Such conversions are common for steroid hormones.

In 1949, when cortisone was found to be a

useful agent in the treatment of arthritis, the

demand for the drug could not be met since no

practical method for large-scale production existed. The chemical synthesis was complicated

and very expensive. In the early and mid-1950’s,

many investigators reported the microbial

transformation of several intermediates to compounds that corresponded to the chemical synthetic scheme. By saving many chemical steps

and achieving higher yields, manufacturers

managed to reduce the price of steroids to a

level where they were a marketable commodity. A conversion of progesterone, for example,

dropped the price of cortisone from $200 to

$6/g in 1949. Through further improvements,

the price dropped to less than $1/g. The 1980

price is $0.46/g.

Developments based on genetic techniques to

increase the production and secretion of key enzymes could substantially improve the economics of some presently inefficient processes. Currently, assessments are being carried out by

various companies to determine which of the

many nonprotein pharmaceuticals can be manufactured more readily or more economically

by biological means.

Approximately 90 percent of the pharmaceuticals used in the treatment of hypertension are

obtained from plants, as well as are miscellaneous cardiovascular drugs. Morphine and

important vasodilators are obtained from the

opium poppy, Papaver somniferum. All these

chemical substances are produced by a series of

enzymes that are coded by corresponding genes

in the whole plant. The long-term possibility

(over 10 years) of using fermentation methods

will depend on identifying the important genes.
Ch. 4—The Pharmaceutical Industry . 77

The genes that are transferred from plant to

bacteria must obviously be determined on a

case-by-case basis. The case study on acetaminophen (the active ingredient in analgesics such as

Tylenol) demonstrates the steps in such a feasibility study. (See app. I-A.)

The first step in such a study is to determine

whether and where enzymes exist to carry out

the necessary transformation for a given product. Acetaminophen for instance, can be made

from aniline, a relatively inexpensive starting

material. The two necessary enzymes can be

found in several fungi. Either the enzymes can

be isolated and used directly in a two-step conversion or the genes for both enzymes can be

transferred into an organism that can carry out

the entire conversion by itself.

Given the cost assumptions outlined in the

case study and the assumptions on the efficiency of converting aniline to acetaminophen, the

cost of producing the drug by fermentation

could be 20 percent lower than production by

chemical synthesis.

Impacts

Genetic technologies can help provide a variety of pharmaceutical products, many of which

have been identified in this report. But the technologies cannot guarantee how a product will

be used or even whether it will be used at all.

The pharmaceuticals discussed have illustrated

the kinds of major economic, technical, social,

and legal constraints that will play a role in the

application of genetic technologies.

Clearly, the major direct impacts of genetic

technologies will be felt primarily through the

type of products they bring to market. Nevertheless, each new pharmaceutical will offer its

own spectrum and magnitude of impacts. Technically, genetic engineering may lead to the production of growth hormone and interferon with

equal likelihood; but if the patient population is

a thousandfold higher for interferon, and if its

therapeutic effect is to alleviate pain and lower

the cancer mortality rate, its impact will be significantly greater.

Many hormones and human proteins cannot

be extensively studied because they are still

either unavailable or too expensive. Until the

physiological properties of a hormone are

understood, its therapeutic values remain unknown. Recombinant DNA techniques are being

used to overcome this circular problem. In one

laboratory, somatostatin is being used as a research tool to study the regulation of the hormonal milieu of burn patients. A single experiment may use as much as 25 mg of the hormone, which, as a product of solid state chemical synthesis, costs as much as $l2,000. Reducing its cost would allow for more extensive

research on its physiological and therapeutic

qualities.

By making a pharmaceutical available, genetic engineering can have two types of impacts.

First, pharmaceuticals that already have medical promise will be available for testing. For example, interferon can be tested for its efficacy

in cancer and viral therapy, and human growth

hormone can be evaluated for its ability to heal

wounds. For these medical conditions, the indirect, societal impact of applied genetics could

be widespread.

Second, other pharmacologically active substances that have no present use will be available in sufficient quantities and at a low enough

cost to enable researchers to explore their possibilities, thus creating the potential for totally

new therapies. Genetic technologies can make

available for example, cell regulatory proteins, a

class of molecules that control gene activity and

that is found in only minute quantities in the

body. The cytokines and lymphokines typify the

countless rare molecules involved in regulation,

communication, and defense of the body to

maintain health. Now, for the first time, genetic

technologies make it possible to recognize, isolate, characterize, and produce these proteins.

The potential importance of this class of pharmaceuticals—the new cell regulatory mole-
78 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

cules—is underscored by the fact that half of

the 22 active INDs for new molecular entities

that have been rated by FDA as promising important therapeutic gains are in the Metabolic

and Endocrine Division, which oversees such

drugs. It is reasonable to anticipate that they

will be employed to treat cancer, to prevent or

combat infections, to facilitate transplantation

of organs and skin, and to treat allergies and

other diseases in which the immune system has

turned against the organism to which it belongs.

(See table 7.)

At the very least, even if immediate medical

uses cannot be found for any of these compounds, their indirect impact on medical research is assured. For the first time, almost any

biological phenomenon of medical interest can

be explored at the cellular level by the applicable 7.–Diseases Amenable to Drugs Produced by

Genetic Engineering in the Pharmaceutical Industry

Drug potentially produced by

Disease or condition genetically engineered organism

Diabetese

Atherosclerosis

Virus diseases

Influenza

Hepatitis

Polio

Herpes

Common cold

Cancer

Anovulation

Dwarfism a

Pain

Wounds and burns

Inflammation,

rheumatic diseasesa

Bone disorders, e.g.,

Paget’s diseasea

Nerve damage

Anemia, hemorrhage

Hemophilia

Blood clotsa

Shocka

Immune disorders

Insulin

Platelet-derived growth factor

(PDGF)

Interferon

Interferon

Hodgkin’s disease

Leukemia

Breast cancer

Human chorionic gonadotropin

Human growth hormone

Enkephalins and endorphins

Human growth hormone

Adrenocorticotrophic hormone

(ACTH)

Calcitonin and parathyroid

hormone

Nerve growth factor (NGF)

Erythropoietin

Factor Vlll and Factor IX

Urokinase

Serum albumin

Cytokines

alndicates dlsaaaes currently treated by the drugs listed.

SOURCE: Office of Technology Assessment.

cation of available scientific tools. These new

molecules are valuable tools for dissecting the

structure and function of the cell. The knowledge gained may lead to the development of

new therapies or preventive measures for

diseases.

The increased availability of new vaccines

might also have serious consequences. But the

extent to which molecular cloning will provide

useful vaccines for intractable diseases is still

unknown. For some widespread diseases, such

as amebic dysentery, not enough is known

about the interaction between the micro-organism and the patient to help researchers design

a rational plan of attack. For others, such as

trachoma, malaria, hepatitis, and influenza,

there is only preliminary experimental evidence

that a useful vaccine could be produced. (See

table 8.) To date, the vaccine that is most likely

to have an immediate impact combats foot-andmouth disease in veterinary medicine. There is

little doubt however, that should any one of the

vaccines for human diseases become available,

the societal, economic, and political consequences of a decrease in morbidity and mortality would be significant. Many of these diseases

are particularly prevalent in less-developed

countries. The effects of developing vaccines

Table 8.-Major Diseases for Which Vaccines

Need To Be Developed

Parasitic diseases

Hookworm

Trachoma

Malaria

Schistosomiasis (river blindness)

Sleeping sickness

viruses

Hepatitis

Influenza

Foot-and-mouth disease (for cloven-hoofed animals)

Newcastle disease virus (for poultry)

Herpes simplex

Mumps

Measles

Common cold rhinoviruses

Varicella-zoster (shingles)

8ecterJa

Dysentery

Typhoid fever

Cholera

Traveller’s diarrhea

SOURCE: Office of Technology Assessment.
Ch. 4—The Pharmaceutical industry ● 79

for them will be felt on an international scale

and will involve hundreds of millions of people.

The new technologies may also lower the

risks of vaccine production. For example, the

FMDV vaccine produced by Genentech is constructed out of 17 of the 20 genes in the entire

virus—enough to confer resistance, but too few

to develop into a viable organism.

The new technology may also supply pharmaceuticals with effects beyond therapy. At least

two promise impacts with broad consequences:

MSH/ACTH 4-10 can be expected to be used on a

wide scale if it is shown to improve memory;

and bombesin and cholecystokinin might expand the appetite suppression market. But neither of these compounds has yet been found to

be useful. While genetic technologies may provide large supplies of the drugs, they do not

guarantee their value.

Antibody-based diagnostic tests, developed

through genetic engineering, may eventually include early warning signals for cancer; they

should be able to recognize any one of the

scores of cancers that cause about a half-million

deaths per year in the United States. If antibodies prove successful as diagnostic screening

agents to predict disease, large-scale screening

of the population can occur, accelerating the

trend toward preventive medicine in the United

States.

In addition to drugs and diagnostic agents,

proteins could be produced for laboratory use.

Expensive, complex media such as fetal calf

serum are presently required for growing most

mammalian tissue cells. Genetic cloning could

make it possible to synthesize vital constituents

cheaply, and could markedly reduce the costs of

cell culture for both research and production.

Ironically, genetic cloning could make economically competitive the very technology that offers an alternative production method for many

drugs: tissue culture.

Nevertheless, the mere availability of a pharmacologically active substance does not ensure

its adoption in medical practice. Even if it is

shown to have therapeutic usefulness, it may

not succeed in the marketplace. Consumer resistance limits the use of some drugs. The American aversion to therapies that require frequent

injection, for instance, is illustrated by the opinion of some that a drug like ACTH offers few, if

any, advantages over steroids.

The use of ACTH is somewhat greater abroad

than in the United States. This is due in part

because physicians in other cultures make far

less use of systemic steroids than their American counterparts, and in part because frequent

injections are more acceptable hence more common. Sales of ACTH in Great Britain—with

its much smaller population—equal American

sales.

At present, the need for injection is a far

more likely deterrent to the wider use of ACTH

than the cost of the drug itself. Reports that it

can be applied by nasal spray suggest that its

use may grow. Implantable controlled-release

dosages may also become available within the

next 5 years. This dependence on appropriate

drug delivery mechanisms may lead to another

line of research–increased attempts to develop

technologies for drug-delivery.

As new pharmaceuticals become available,

disruption can be expected to occur in the supply of some old ones. Pharmaceuticals whose

production is tied to the production of others

might become increasingly expensive to produce. Clotting factors, for example, are extracted with other blood components from

plasma. Nevertheless, producing any of the 14

currently approved blood plasma products by

rDNA would reduce the incidence of hepatitis

caused by contamination from natural blood

sources.

Whether new pharmaceuticals are produced

or new production methods for existing pharmaceuticals are devised, future sources for the

drugs may change. Currently, the sources are

diverse, including many different plants, numerous animal organs, various tissue culture

cells, and a wide range of raw materials used

for chemical synthesis. A massive shift to fermentation would narrow the selection. The impacts on present sources can only be judged on

a case-by-case basis. The new sources—microorganisms and the materials that feed them—

offer the guarantee that the raw materials won’t

dry up. If one disappears, another can be found.
80 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Clearly, there is no simple formula to identify Given the assumptions described, the immediall the impacts of applied genetics on the phar- ate direct economic impact of using genetic mamaceutical industry. Even projections of eco- nipulation in the industry, measured as sales,

nomic impacts must remain crude estimates. can be estimated in the billions of dollars, with

Nevertheless, the degree to which genetic engi- the indirect impacts (sales for suppliers, savings

neering and fermentation technologies might due to decreased sick

potentially account for drug production in spe- several times that value.

cific categories is projected in appendix I-B.

days, etc.) reaching

Technical notes

1. Many hormones are simply chains of amino acids (poly -

peptides); some are polypeptides that have been modified by the attachment of carbohydrates (glycopeptides). Hormones usually trigger events in cells remote

from the cells that produced them. Some act over

relatively short distances—between segments in the

brain, or in glands closely linked to the brain, others

act on distant sites in tissues throughout the body.

2. For peptides about 30 AA in length, the cost may ap3.

4.

preach $1 per mg as the volume approaches the kilogram level–a level of demand rarely existing today but

likely to be generated by work in progress. Today, the

cost of the 32 AA polypeptide, calcitonin, which is synthesized chemically and marketed as a pharmaceutical

product by Armour, is probably in the range of $20 per

mg, since the wholesale price in vials containing approximately 0.15 mg is about $85/mg. (That price is an

educated guess, since such costs are closely guarded

secrets and since the price of a pharmaceutical includes so many variables that the cost of the agent

itself is a small consideration. )

In addition to those helped by the National Pituitary

Agency, another 100 to 400 patients are treated with

hGH from comercial sources. The commercial price

is approximately $15 per unit (roughly $30/mg). The

production cost at the National Pituitary Agency is

about $0.75/unit ($1.50/mg). The National Pituitary

Agency produces 650,000 international units (lU)

(about 325 g) of hGH, along with the thyroid-stimulating hormone, prolactin, and other hormones, from

about 60,000 human pituitaries collected each year.

That is enough hGH both for the current demand and

for perhaps another 100 hypopituitary patients.

Workers at the Howard Hughes Medical Institute of

the University of California, San Francisco, isolated

messenger RNA from a human pituitary tumor and

converted it into a DNA-sequence that could be put into

E. coli. The sequence, however, was a mixture of hGH

and non-hGH material. It has been reported that Eli Lilly & Co., which has provided some grant money to the

Institute, has obtained a license to the patents relating

to this work. Grants from the National Institutes of

Health and the National Science Foundation were also

acknowledged in the publication.

At practically the same time, researchers at Genentech, in conjunction with their associates at City of

Hope National Medical Center disclosed the production

of an hGH analog. This was the first time that a human

polypeptide was directly expressed in E. coli in functional form. The work was supported by Kabi Gen AB,

and Kabi has the marketing rights.

The level of hGH production reported in the scientific account of the Genentech work was on the same

order as that reported for the insulin fragments—

approximately 186,000 hGH molecules per cell—a level

that might be competitive even before efforts are made

to increase yield. Genentech stresses the point that design, rather than classical mutation and selection, is the

logical way to improve the system, since the hormone’s

“blueprint” is incorporated in a plasmid that can be

moved between strains of E. coli, between species, or

even from simple bacteria into more complex organisms, such as yeast.

5. Since erythropoietin is a glycoprotein, it may not be

feasible to synthesize the active hormone with presently available rDNA techniques.

6. Antigens are surface components of pathogenic organisms, toxins, or other proteins secreted by pathogenic

micro-organisms. They are also the specific counterparts of antibodies: antibodies are formed by the

body’s immune system in response to their presence.

Antibodies are synthesized by white blood cells and are

created in such a way that they are uniquely structured to bind to specific antigens.

7. Many of the most devastating infectious diseases involve complex parasites that refuse to grow under laboratory conditions. The first cultivation of the most

malignant of the species of protozoa that causes malaria, using human red blood cells, was described in

1976 by a Rockefeller University parasitologist, William

Rager. Experimental immunogens were prepared and

showed promise in monkeys, but concern about the existence of the red blood cell remnants—which could

give rise to autoimmune reactions–curtailed the prospect for making practical vaccines by that route. Several biotechnology firms are currently trying to synthesize malaria antigens by molecular cloning. This effort

may produce technical solutions to such scourges as
Ch. 4—The Pharmaceutical industry ● 81

schistosomiasis (bilharzia), filariasis (onchocerciasis

and elephantiasis), Ieshmaniasis, hookworm, amebic infections, and trypanosomiasis (sleeping sickness and

Chagas’ disease).

8. Another potential use of antigens is suggested by the

experimental treatment of stage I lung cancer patients

with vaccines prepared from purified human lung

cancer antigens, which appears to substantially prolong survival. And the Salk Institute is expanding clinical trials in which a procine myelin protein prepared

by Eli Lilly & Co. is injected into multiple sclerosis patients to mop up the antimyelin antibodies that those

patients are producing. Fifteen to forty-two g of myelin

have been injected without adverse effects, suggesting

a new therapeutic approach to auto-immune diseases.

The protein appears to suppress the symptoms of experimental allergic encephalomyelitis, an animal disease resembling multiple sclerosis. Should this research succeed, the use of molecular clones to produce

human protein antigens seems inevitable.

9. There are at least two distinct kinds of “classical” interferons—leukocyte interferon and fibroblast interferon,

so-called for the types of cells from which they are obtained. A third kind, called lymphoblastoid because it is

produced from cells derived from a Burkitt’s lymphoma, appears to be a mixture of the other two interferon. All produce the antiviral state and are induced

by viruses. A fourth kind, known as “immune” interferon, is produced by lymphocytes. Some evidence indicates that it may be a more potent antitumor agent

than the classical types. Currently, interferon is obtained chiefly from white blood cells (leukocytes) from

the blood bank in Helsinki that serves all of Finland, or

from fibroblasts grown in cell culture.

10. Recently, G. D. Searle & Co. announced that new technology developed at its R&D facility in England has increased the yield of fibroblast interferon by a factor of

60. On the basis of this process, Searle expects to supply material for the first large-scale clinical trial of

fibroblast interferon. Abbott Laboratories also recently

announced plans to resume production of limited

quantities of fibroblast interferon for clinical studies it

plans to sponsor.

Unlike leukocytes and specially treated fibroblasts,

which can be used only once, lymphoblasts derived

from the tumor Burkitt’s lymphoma grow freely in

suspension and produce the least costly interferon

presently obtainable. However, they also produce a disadvantageous mixture of both leukocyte and fibroblast

interferon. The Burroughs-Wellcome Co. produces

lymphoblastoid interferon in 1,000-1 fermenters and

has begun clinical trials in England, but the U.S. FDA

has generally resisted efforts to make use of products

derived from malignant cells. It is used extensively in

research, and FDA is considering evidence from Burroughs-Wellcome that may lead to a relaxation of the

prohibition, under pressure from the National Cancer

Institute.

11. What may be a landmark patent has been issued to

Hilary Koprowski and Carlos Croce of the Wistar Institute (for work done under the then Department of

Health, Education, and Welfare funding) on the production of monoclinal antibodies against tumor cells.

In a number of examples, these researchers demonstrated that an animal can be immunized with tumor

cells, and that hybridomas derived from that animal

will produce antibodies that demonstrate a specificity

for the tumor.

The final sentence of the patent text provides the rationale for the use of antibodies in both cancer and infectious disease therapies: “If the (tumor) antigen is

present, the patient can be given an injection of an antibody as an aid to react with the antigen. ” (U.S.

4,172,124.)

12. Myeloma cells grow vigorously in culture and have the

unique characteristic of producing large quantities of

antibodies. Each spleen cell of the immune type, on the

other hand, produces an antibody that recognizes a

single antigen, but these do not grow well in culture.

When normal immune spleen cells are fused with myeloma cells, the resulting mixture of genetic capacities

forms a cell, called a “hybridoma,” which displays the

desired characteristics of the parent cells: 1) it secretes

the antibody specified by the genes of the spleen cell;

and 2) it displays the vigorous growth, production, and

longevity that is typical of the myeloma cell.

13. The use of high-correlation antibody assays in cancer

studies has only just begun. Antibodies that have been

treated so they can be seen with X-rays and that are

specific for a tumor, can be used early to detect the occurrence or spread of tumor cells in the body. Because

some 785,000 new cancer cases will be detected in

1980 with current diagnostic methods, because cancer

will cause 405,000 deaths, and because early detection

is the major key to improving survival, the implications

are indeed enormous.

14. In the late 1950’s, Lederle Laboratories marketed a

preparation of 95-percent pure streptokinase (a bacterially produced enzyme that dissolves blood clots) for

intravenous administration. They withdrew the product from the market around 1960 because it caused

allergic reactions, which dampened clinical enthusiasm

for its therapeutic potential.

The presence in human urine of urokinase, an enzyme also capable of removing blood clots, was also discovered in the early 1950’s. Urokinase was purified,

crystallized, and brought into clinical use in the mid1960’s. From the beginning it was apparent that “an intense thrombolytic state could be achieved with a

much milder coagulation defect than occurred with

streptokinase; no pyrogenic or allergic reactions were

noted, and no antibodies resulted from its administration . . . There did not appear to be as great variation in

patient responsiveness.” In 1967-68 and 1970-73, the

National Heart and Lung Institute organized clinical

trials that compared urokinase with streptokinase and

heparin, an anticoagulant, in the treatment of pulmonary embolism. The trials indicated that streptokinase and urokinase were equivalent and superior

to heparin over the short term, although their long-
82 ● Impacts of Applied Genetics —Micro-Organisms, Plants, and Animals

term benefits were not established. Since then, clinical

investigation of urokinase has been hampered by

domestic regulatory problems, which have raised the

cost of production and restricted its availability in the

United States.

In January 1978, Abbott Laboratories obtained a

new drug application for urokinase and introduced the

product Abbokinase; by that time, however, the sales

of urokinase in Japan were already pushing $90 million

per year. Recently, Sterling Drug has begun marketing

a urokinase product (Breokinase) manufactured by

Green Cross of Japan: “According to Japanese reports,

urokinase is the first Japanese-made drug formulation

to receive production and sales approval from FDA.

Green Cross estimates that within 3 years of the start

of Sterling’s marketing activities, the value of urokinase exports will reach Yen 500 million ($2.12 million) per month, and considers that its profits from exporting a finished product will probably be better than

those from bulk drug sales or the licensing of technology. ” The Green Cross product is made from human

urine collected throughout Korea and Japan, and takes

advantage of technology licensed from Sterling. Abbott’s product, on the other hand, is derived from

kidney-cell culture.

15. Intergeneric hybrids have extremely interesting possibilities. For example, it would be beneficial to cephaIosporin-process technology to combine in one organism the acyltransferase from Penicillium chrysogenum

and the enzymes of C. acremonium, which does not incorporate side chain precursors onto cephalosporin

like P. chrysogenum does for penicillins.

16. Another example of recombination between species is

that reported for two species of fungi, Aspergillus nidulans and A. rugulosus, subsequent to protoplasm fusion.

The only report of a successful cross between

genera using protoplasm fusion technology has been between the yeasts Candida tropicalis and Saccariomycopsis fibuligera, which took place at low frequency

and gave rise to types intermediate between the

parents.

17. An example of screening is provided by the new

B-lactam (penicillin-like) antibiotics. Using older

screening methods, no new B-lactams were found

from 1956 until 1972 when a new method was devised.

A new series of these antibiotics was thus found.

Within the past year, 6 new B-lactams have been

commercialized and at least 12 more are in clinical

trials around the world. The sales forecasts for these

new agents are estimated to be over $1 billion. 
Chapter 5

The Chemical Industry
Chapter 5

Page

Background . . . . . . . . . . . . . . . . . . . . . . . . . 85

Overview of the Industry . . . . . . . . . . . . . . . . . . 85

Fermentation and the Chemical Industry. . . . . 87

New Process Introduction. . . . . . . . . . . . , , . . . . 89

Characteristics of Biological Production

Technologies. . . . . . . . . . . . . . . . . . . . . . . . . 90

Renewable Resources. . . . . . . . . . . . . . . . . . 90

Physically Milder Conditions. . . . . . . . . . . . . 91

One-Step Production Methods. . . . . . . . . . . . 91

Reduced Pollution... . . . . . . . . . . . . . . . . . 91

Industrial Chemicals That May Be Produced by

Biological Technologies. . . . . . . . . . . . . . . . . 92

Fertilizers, Polymers, and Pesticides. . . . . . . . . 94

Constraints on Biological Production Techniques 96

An Overview of Impacts . . . . . . . . . . . . . . . . . . . 97

Impacts on Other Industries. . . . . . . . . . . . . . . 97

Impacts on University Research. . . . . . . . . . . . 98

The Social Impacts of Local Industrial Activity . 99

Impacts on Manpower. . . . . . . , . . . . . . . . . . . 99

Tables

Table No. Page

9. Data for Commercially Produced Amino

Acids. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88

10.

11.

12.

13.

14.

15.

16.

17.

18.

19.

Summary of Recent Estimates of Primary

U.S. Cost Factors in the Production of 

Lysine Monohydrochloride by Fermentation

and Chemical Synthesis. . . . . . . . . . . .

Some Commercial Enzymes and Their U

Expansion of Fermentation Into the

Chemical Industry, . . . . . . . . . . . . . . .

The Potential of Some Major Polymeric

Materials for Production Using Biotechnology 95

Some Private Companies With

Biotechnology Programs ., . . . . . . . . . . . . , . 98

Distribution of Applied Genetics Activity

in Industry . . . . . . . . . . . . . . . , . . . . . . . . . . 100

Manpower Distribution of a Firm With

Applied Genetics Activity. . . . . . . . . . . . . . . . 100

Index to Fermentation Companies. . . . . . . . . 100

Fermentation Products and Producers . . . . . 101

U.S. Fermentation Companies . . . . . . . . . . . . 103

Figures

Figure No. Page

24. Flow of industrial Organic Chemicals From

Raw Materials to Consumption . . . . . . . . . . . 86

25. Diagram of Alternative Routes to Organic

Chemicals , . . . . . . . . . . . . . . . . . . . . . . . . . . 90
Chapter 5

The Chemical Industry

Background

The organic substances first used by humans

to make useful materials such as cotton, linen,

silk, leather, adhesives, and dyes were obtained

from plants and animals and are natural and renewable resources. In the late 19th century,

coal tar, a nonrenewable substance, was found

to be an excellent raw material for many organic compounds, When organic chemistry developed as a science, chemical technology improved. At about the same time relatively cheap

petroleum became widely available. The industry shifted rapidly to using petroleum as its major raw material.

The chemical industry’s constant search for

cheap and plentiful raw materials is now about

to come full circle. The supply of petroleum,

which presently serves more than 90 percent of

the industry’s needs, is severely threatened by

both dwindling resources and increased costs. It

has been estimated that at the current rate of

consumption, the world’s petroleum supplies

will be depleted in the middle of the next century. Most chemical industry analysts, therefore, foresee a shift first back to coal and then,

once again, to the natural renewable resources

referred to as biomass. The shifts will not

necessarily occur sequentially for the entire

Overview of the industry

The chemical industry is one of the largest

and most important in the world today. The U.S.

market for synthetic organic chemicals alone,

excluding primary products made from petroleum, natural gas, and coal tar, exceeded $35

billion in 1978.

The industry’s basic function is to transform

low-cost raw materials into end-use products of

greater value. The most important raw materials are petroleum, coal, minerals (phosphate,

carbonate), and air (oxygen, nitrogen). Roughly

two-thirds of the industry is devoted to producchemical industry. Rather, both coal and biomass will be examined for their potential roles

on a product-by-product basis. 1

The chemical industry is familiar with the

technology of converting coal to organic chemicals, and a readily available supply exists. Coalbased technologies will be used to produce a

wide array of organic chemicals in the near future. * Nevertheless, economic, environmental,

and technical factors will increase the industry’s

interest in biomass as an alternative source for

raw materials. Applied genetics will probably

play a major role in enhancing the possibilities

by allowing biomass and carbohydrates from

natural sources to be converted into various

chemicals. Biology will thereby take on the dual

role of providing both raw materials and a process for production.

‘For further details see Ener,qv From Biologica/ Processes, ~wl. 1,

OTA.E-124 (Washington, D. C.: office of ‘rechnolo~v Assessment,

July 1980).

*Most important organic intermediiites (chemical compounds

used fol. the illdustl’iitl s.vnthesis of c~n~n]et.ciiil produ(’ts SU(’11 iis

pliist i(:s itnd fihers) can be obtained from coal as an iilterniit i\(l Iiiii

nliiteriiil. (kirrentl. v, methods iire heing dtn’elopecl to (’onirrt {’oiil

into “s.vnthetic gas, ” which (;i]n then h{’ used ils ]’ii}~ l]]i~t[>}”ii]l fo].

turther conivwions.

ing inorganic chemicals such as lime, salt, ammonia, carbon dioxide, chlorine gas, and hydrochloric and other acids.

The other third, which is the target for biotechnology, produces organic chemicals. Its output includes plastics, synthetic fibers, organic

solvents, and synthetic rubber. (See figure 24.)

In general, petroleum and natural gas are first

converted into “primary products” or basic organic chemicals such as the hydrocarbons ethylene and benzene. These are then converted into

a wide range of industrial chemicals. Ethylene

85

76-565 0 - 81 - 7
86 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Figure 24.-Flow of Industrial Organic Chemicals From Raw Materials to Consumption

Organic resources

80% raw material from petroleum/

natural gas

20°/0 raw material from coal, coke, and

renewable resources

SOURCE: U.S. Industrh?l Outlook (W.shlngton, D. C.: Department of Commerce, 1978); Kline Gu/de to Chemioel Industry, Fairfield,

N.J., adapted from Tong, 1979.
Ch. 5—The Chemical industry ● 87

alone serves as the basic chemical for the manufacture of half of the largest volume industrial

chemicals. Each of the steps in a chemical conversion process is controlled by a separate reaction, which is often performed by a separate

company.

Evaluating the competitiveness both of a

process and of the market is critical for the

chemical industry, which is intensive for capital, energy, and raw materials. Its plants use

large amounts of energy and can cost hundreds

of millions of dollars to build, and raw material

costs are generally 50 to 80 percent of a product’s cost. If a biological process can use the

same raw materials and reduce the process cost

by even 20 percent, or allow the use of inexpensive raw materials, it could provide the industry

with a major price break.

Fermentation and

the chemical industry

The production of industrial chemicals by

fermentation is not new. Scores of chemicals

have been produced by micro-organisms in the

past, only to be replaced by chemical production based on petroleum. In 1946, for example,

27 percent of the ethyl alcohol in the United

States was produced from grain and grain products, 27 percent from molasses, a few percent

each from such materials such as potatoes, pineapple juice, cellulose pulp, and whey, and only

36 percent from petroleum. Ten years later

almost 60 percent was derived from petroleum.

Even more dramatically, fumaric acid was at

one time produced on a commercial scale

through fermentation, but its biological production was stopped when a more economical synthesis from benzene was developed. Frequently,

after a fermentation product was discovered,

alternative chemical synthetic methods were

soon developed that used inexpensive petroleum as the raw material.

Nevertheless, for the few chemical entities

still produced by fermentation, applied genetics

has contributed to the economic viability of the

process. The production of citric and lactic

acids and various amino acids are among the

processes that have benefited from genetics.

Lactic acid is produced both synthetically and

by fermentation. Over the past 10 to 20 years,

manufacture by fermentation has experienced

competition from chemical processes.

The organisms used for the production of lactic acid are various species of the bacterium Lactobacillus. Starting materials may be glucose, sucrose, or lactose (whey). The fermentation per

se is efficient, resulting in 90 percent yields, depending on the original carbohydrate. Since

most of the problems in the manufacture of lactic acid lie in the recovery procedure and not in

fermentation, few attempts have been made to

improve the industrial processes through

genetics.

Citric acid is the most important acidulant,

and historically has held over 55 to 65 percent

of the acidulant market for foods. * It is also

used in pharmaceuticals and miscellaneous industrial applications. It is produced commercially by the mold Aspergillus niger. Surprisingly little work has been published on improving citric

acid-producing strains of this micro-organism.

Weight yields of 110 percent have recently been

reported in A. niger mutants obtained by irradiating a strain for which a maximum yield of

29 percent had been reported.

Amino acids are the building blocks of proteins. Twenty of them are incorporated into

proteins manufactured in cells, others serve

specialized structural roles, are important metabolic intermediates, or are hormones and neurotransmitters. All of the amino acids are used

in research and in nutritional preparations,

with most being used in the preparation of

pharmaceuticals. Three are used in large quantities for two purposes: glutamic acid to manufacture monosodium glutamate, which is a fla-

● The other two important acidukmts, or acidifying agents, are

phosphoric acid (20 to 25 percent) and maltc acid (S percent),
88 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

vor enhancer particularly in oriental cooking;*

and lysine and methionine as animal feed additives.

Conventional technology for producing glutamic acid is based on pioneering work that was

subsequently applied to other amino acids. The

production employed microbial strains to produce amino acids that are not within their normal biosynthetic capabilities. This was accom-

“Monosodium glutamate is the sodium salt of glutamic acid. In

1978, about 18,000 tonnes were manufactured in the United

States and about 11,000 tonnes imported. The food industry consumed 97 percent. The fermentation plant of the Stauffer Chemical Co. in San Jose, Calif., is the sole U.S. producer. The microbes

used in glutamic acid fermentation (Corynebacferium glutamicum,

C. Iileum, and Brevibacterium j?avum) produce it in 60 percent of

theoretical yield. Thus, there is some but not great potential for

the use of applied genetics to improve the yield. Many of the genetic approaches have already been thoroughly investigated by industrial scientists.

plished by using two methods: I) manipulating

microbial growth conditions, and 2) isolating

nat urally occurring mutants.

Although microbial production of all the

amino acids has been studied, glutamic acid and

L-lysine* * are the ones produced in significant

quantities by fermentation processes. (See table

9.) The production of L-lysine is an excellent ex-

“ *The lack of a single amino acid can retard protein synthesis,

and therefore growth, in a mammal. The limiting amino acid is a

function of the animal and its feed. The major source of animal

feed in the United States is soybean meal. The limiting amino acid

for feeding swine is lysine, the limiting amino acid for feeding

poultry is methionine. Because of increased poultry demand,

world demand for Iysine is climbing, Eurolysine is spending $27

million to double its production capacity in Amiens, France, to 10

thousand tonnes. The Asian and Mideast markets are ;stimated to

increase to 3 thousand tonnes in 1985. Some bar .eria produce

lysine at over 90 percent of theoretical yield, L:[tle genetic improvement is likely in this conversion yield, however, significant

improvement can be made in the rate and final concentration.

Table 9.—Data for Commercially Produced Amino Acidsa

Price March Potential for application of

1980 (per kg Production 1978 biotechnology (de novo synthesis or

Amino acid pure L) Present source (tonnes) bioconversion; organisms and enzymes)

Alanine. ... .. ... ... ... ... .$ 80

Arginine . . . . . . . . . . . . . . . . . . .

Asparagine. . . . . . . . . . . . . . . . .

Aspartic acid . . . . . . . . . . . . . . .

Citrulline. . . . . . . . . . . . . . . . . . .

Cysteine. . . . . . . . . . . . . . . . . . .

Cystine. . . . . . . . . . . . . . . . . . . .

DOPA (dihydrophenylalanine) .

Glutamic. . . . . . . . . . . . . . . . . . .

Glutamine. . . . . . . . . . . . . . . . . .

Histidine. . . . . . . . . . . . . . . . . . .

Hydroxyproline . . . . . . . . . . . . .

Isoleucine. . . . . . . . . . . . . . . . . .

Leucine. . . . . . . . . . . . . . . . . . . .

Lysine. . . . . . . . . . . . . . . . . . . . .

Methionine. . . . . . . . . . . . . . . . .

Ornithine . . . . . . . . . . . . . . . . . .

Phenylalanine . . . . . . . . . . . . . .

Proline . . . . . . . . . . . . . . . . . . . .

Serine. . . . . . . . . . . . . . . . . . . . .

Threonine. . . . . . . . . . . . . . . . . .

Tryptophan. . . . . . . . . . . . . . . . .

Tyrosine. . . . . . . . . . . . . . . . . . .

Valine . . . . . . . . . . . . . . . . . . . . .

28

50

12

250

50

60

750

4

55

160

280

350

55

350

265

60

55

125

320

150

110

13

60

Hydrolysis of protein; 10-50(J) b 

–

chemical synthesis

Gelatin hydrolysis 200-300 (J) Fermentation in Japan

Extraction 10- 50(J) –

Bioconversion of

fumaric acid 500-1,000 (J) Bioconversion

10-90 (J) Fermentation in Japan

Extract ion 100- 200(J) –

Extraction 100- 200(J) –

Chemical 100- 200(J) —

Fermentation 10,000-100,000 (J) De novo: Micrococcus gutamicus

Extraction 200-300 (J) Fermentation in Japan

— 100-200 Fermentation in Japan

Extraction from collagen 10-50 —

Extraction

—

Fermentation (800A)

Chemical (20%)

Chemical from acrolein

—

Chemical from

benzaldehyde

Hydrolysis of gelatin

—

—

Chemical from indole

Extraction

—

aproduction data largely from Japan because of relative Small U.S. production. L

10-50 (J)

50-100(J) Fermentation in Japan

10,000 (J) (80% by fermentation) De novo:

Corynebacterium glutamicum and

Brevibacterium flavum

17,000(D,L) c —

20,000 (D,L) (J)

10-50 (J) Fermentation in Japan

50-100 (J) Fermentation in Japan

10-50 (J) Fermentation in Japan

10-50 (J) Bioconversion in Japan

50-10 (J) Fermentation in Japan

55 (J)

50- 100(J) -

50-100 (J) Fermentation in Japan

‘Japan.

CD and L forms.

SOURCE: Massachusetts Institute of Technology.
Ch. 5—The Chemical Industry . 89

ample of the competition between chemical and mostly from Japan and South Korea. Recent

biotechnological methods. Fermentation has estimates of primary U.S. cost factors in the

been gradually replacing its production by competing production methods are summarized

chemical synthesis; in 1980, 80 percent of its in table 10. Fermentation costs are lower for all

worldwide production is expected to be by mi- three components of direct operating costs:

crobes. It is not produced in the United States, labor, material, and utilities.

which imported about 7,000 tonnes in 1979,

Table 10.—Summary of Recent Estimates of Primary U.S. Cost Factors in the Production of

L= Lysine Monohydrochloride by Fermentation and Chemical Synthesis

Cost factors in Production of 98% L-lysine monohvdrochloride

By fermentation By chemical synthesis

Requirement Estimated 1976 cost Requirement Estimated 1976 cost

(units per unit per unit product (units per unit per unit product

(product) Cents/lb Cents/kg product) Cents/lb Cents/lb

Total Iaborc. . . . . . . . . . . . . . . . . — 8 18 — 9 20

Materials

Molasses . . . . . . . . . . . . . . . . 44 7 16 —

Soy beanmeal, hydrolyzed. . .

— —

0.462 4 9 —

Cyclohexanol. . . . . . . . . . . . . —

— —

— 0.595 17 37

Anhydrous ammonia. . . . . . . — — — 0.645 6 14

Other chemicalsd. . . . . . . . . . — 7 15 — 4 10

Nutrients and solvents. . . . . — — — — 4 8

Packaging, operating, and

maintenance materials. . . 10 22 — 9 21

Total materials. . . . . . . . . . — 28 62 — 45 90

Total utilities . . . . . . . . . . . . . . — 6 12 — 7 16

Total direct operating cost — 42 92 — 56 126

Plant overhead, taxes,

and insurance . . . . . . . . — 10 21 — 10 21

Total cash cost . . . . . . . . . — 52 11 — 66 147

Depreciation. . . . . . . . . . . — 16 35 — 13 28

Interest on working capital — 1 3 — 1 3

Total costg . . . . . . . . . . . — 69 151 — 80 178

aAggumes a 23-percent yield on molasses.

bAssumes a 65-percent yield on cyclohexanol.

clncludes operating, maintenance, and control laboratory labor.

dFor both the proce~~ of fermentation and chemical synthesis, assumed use of hydrochloric acid (~ percent) and ammonia (n percent). For fermentation includes dSO

potassium diphosphate, urea, ammonium suifate, calcium carbonate, and magnesium sulfate. For chemical synthesis also includes nitrosyl chloride, sulfuric acid,

and a credit for ammonium sulfate byproduct.

~otal utiiities for both processes include cooling water, steam process water, and electricity. For chemical synthesia, natural gas is also included.

fTen percent ~r year of fixed capital costs for a new 20 million lb per year U.S. plant built in 1975 at assumed capital cost of $36.6X 10’ for fOrmOntatiOn and $32.5x 10’

for chemical synthesis exclusive of land costs.

SOURCE: Stanford Research Institute, Chemical Economics Handbook 563:3401, May 1979.

New process introduction

The development of biotechnology should be

viewed not so much as the creation of a new industry as the revitalization of an old one. Both

fermentation and enzyme technologies will

have an impact on chemical process development. The first will affect the transition from

nonrenewable to renewable raw materials. The

second will allow fermentation-derived products to enter the chemical conversion chains,

and will compete directly with traditional chemical transformations. (See figure 25. ) Fermentation, by replacing various production steps,

could act as a complementary technology in the

overall manufacture of a chemical.
90 ● Impacts of Applied Genetics-Micro-Organisms, Plants,and Animals

Figure 25.—Diagram of Alternative Routes to Organic Chemicals

Petroleum Carbohydrates

% followed by number indicatea iength of carbon chain.

SOURCE: (3. E. Tong, “industrial Chemicals From Fermentation Enzymes,” M/croLr. TechrroL, voi. 1,1979, PP. 173-179.

Characteristics of biological

production technologies

The major advantages of using commercial

fermentation include the use of renewable resources, the need for less extreme conditions

during conversion, the use of one-step production processes, and a reduction in pollution. A

micro-organism might be constructed, for example, to transform the cellulose in wood directly into ethanol. * (App. 1-D, a case study of

the impact of genetics on ethanol production,

elaborates these points. )

RENEWABLE RESOURCES

Green plants use the energy captured from

sunlight to transform carbon dioxide from the

“If IWILI(WI I“OIS iil)~)]S()\’i~l 01 SU(SI1 illl it(s(:oilll)lislllllel~l I)$v I’DNA

Icchniqut?s wits sulmlilted to lht> Ik:otl]l)iiliii)l DNA Advisor y

(kmlnlilltw ill the %?pl. 25, 1980 mt?eling.

atmosphere into carbohydrates, some of which

are used for their own energy needs. The rest

are accumulated in starches, cellulose, lignins,

and other materials called the biomass, which is

the foundation of all renewable resources.

The technologies of genetic engineering could

help ease the chemical industry’s dependence

on petroleum-based products by making the use

of renewable resources attractive. All microorganisms can metabolize carbohydrates and

convert them to various end products. Extensive research and development (R&D) has

already been conducted on the possibility of

using genetically engineered strains to convert

cellulose, the major carbohydrate in plants, to

commercial products. The basic building block

of cellulose—glucose—can be readily used as a

raw material for fermentation.
Ch. 5—The Chemical industry ● 91

other plant carbohydrates include cornstarch, molasses, and lignin. The last, a polymer

found in wood, could be used as a precursor for

the biosynthesis of aromatic (benzene-like)

chemicals, making their production simpler and

more economical. Nevertheless, the increase in

the price of petroleum is not a sufficient reason

for switching raw materials, since the cost of

carbohydrates and other biological materials

has been increasing at a relative rate.

PHYSICALLY MILDER CONDITIONS

In general, there are two main ways to speed

chemical reactions: by increasing the reaction

temperature and by adding a catalyst. A catalyst

(usually a metal or metal complex) causes one

specific reaction to occur at a faster rate than

others in a chemical mixture by providing a surface on which that reaction can be promoted.

Even using the most effective catalyst, the conditions needed to accelerate industrial organic

reactions often require extremely high temperatures and pressures—several hundred degrees Celsius and several hundred pounds per

square inch.

Biological catalysts, or enzymes, on the other

hand can speed-up reactions without the need

for such extreme conditions. Reactions occur in

dilute, aqueous solutions at the moderate conditions of temperature, pressure, and pH (a measure of the acidity or alkalinity of a solution) that

are compatible with life.

ONE-STEP PRODUCTION METHODS

In the chemical synthesis of compounds, each

reaction must take place separately. Because

most chemical reactions do not yield pure products, the product of each individual reaction

must be purified before it can be used in the

next step. This approach is time-consuming and

expensive. If, for example, a synthetic scheme

that starts with ethylene (a petroleum-based

product) requires 10 steps, with each step yielding 90 percent product (very optimistic yields in

chemical syntheses), only about one-third of the

ethylene is converted into the final end product.

Purification may be costly; often, the chemicals

involved (such as organic solvents for extractions) and the byproducts of the reaction are

toxic and require special disposal.

In biological systems, micro-organisms often

complete entire synthetic schemes. The conversion takes place essentially in a single step,

although several might occur within the organisms, whose enzymes can transform the precursor through the intermediates to the desired

end product. Purification is not necessary.

REDUCED POLLUTION

Metal catalysts are often nonspecific in their

action; while they may promote certain reactions, their actions are not ordinarily limited to

making only the desired products. Consequently, they have several undesirable features: the

formation of side-products or byproducts; the

incomplete conversion of the starting material(s); and the mechanical and accidental loss of

the product.

The last problem occurs with all types of synthesis. The first two represent inefficiencies in

the use of the raw materials. These necessitate

the separation and recycling of the side-products formed, which can be difficult and costly

because they are often chemically and physically similar to the desired end products. (Most

separation techniques are based on differences

in physical properties—e.g., density, volatility,

and size.)

When byproducts and side-products have no

value, or when unconverted raw material cannot be recycled economically, problems of

waste disposal and pollution arise. Their solution requires ingenuity, vigilance, energy, and

dollars. Many present chemical processes create

useless wastes that require elaborate degradation procedures to make them environmentally

acceptable. In 1980, the chemical industry is expected to spend $883 million on capital outlays

for pollution control, and well over $200 million

on R&D for new control techniques and replacement products. These figures do not include the millions of dollars that have been

spent in recent years to clean up toxic chemical

dumps and to compensate those harmed by

poorly disposed wastes, nor do they include the

cost of energy and labor required to operate

pollution-control systems.

A genetically engineered organism, on the

other hand, is designed to be precursor- and
92 . Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

product-specific, with each enzyme having

essentially 100-percent conversion efficiency.

An enzymatic process that carries out the same

transformation as a chemical synthesis produces no side-products (because of an enzyme’s

high specificity to its substrate) or byproducts

(because of an enzyme’s strong catalytic power).

Consequently, biological processes eliminate

many conventional waste and disposal problems

at the front end of the system-in the fermenter. This high conversion efficiency reduces

the costs of recycling. In addition, the efficiency

of the biological conversion process generally

simplifies product recovery, reducing capital

and operating costs. Furthermore, by their

nature, biologically based chemical processes,

tend to create some waste products that are biodegradable and valuable as sources of nutrients.

Specific comparisons of the environmental

hazards produced by conventional and biological systems are difficult. Data detailing the

pollution parameters for various current chemical processes exist, but much less information

is available for fermentation processes, and few

Industrial chemicals that

by biological technologies

Despite the benefits of producing industrial

chemicals biologically, thus far major fermentation processes have been developed primarily

for a few complex compounds such as enzymes.

(See table 11.) Biological methods have also been

developed for a few of the simpler commodity

chemicals: ethanol, butanol, acetone, acetic

acid, isopropanol, glycerol, lactic acid, and citric

acid.

Two questions are critical to assessing the

feasibility or desirability of producing various

chemicals biologically:

I. Which compounds can be produced biologically (at least theoretically)?

2. Which compounds may be primarily dependent on genetic technology, given the

costs and availability of raw materials?

compounds are produced by both methods.

However, in most beverage distilling operations,

pollution has been reduced to almost zero with

the complete recovery of still slops as animal

feeds of high nutritional value. Such control

procedures are generally applicable to most

fermentation processes. (App. I-C describes the

pollutants that may be produced by current

chemical processes and those expected from

biologically based processes.)

The Environmental Protection Agency has

estimated that the U.S. Government and industry combined will spend over $360 billion to

control air and water pollution in the decade

from 1977 through 1986. The share of the

chemical and allied industries is about $26 billion. Genetic engineering technology may help

alleviate this burden by offering cleaner processes of synthesis and better biological waste

treatment systems. The monetary savings could

be tremendous. As pure speculation, if just 5

percent of the current chemical industry were

affected, spending on pollution could be reduced by about $100 million per year.

may be produced

In principle, virtually all organic compounds

can be produced by biological systems. If the

necessary enzyme or enzymes are not known to

exist, a search of the biological world will probably uncover the appropriate ones. Alternatively, at least in theory, an enzyme can be

engineered to carry out the required reaction.

Within this framework, the potential appears to

be limited only by the imagination of the biotechnologist—even though certain chemicals

that are highly toxic to biological systems are

probably not amenable to production.

Three variables in particular affect the

answer to the second question: the availability

of an organism or enzymes for the desired

transformation; the cost of the raw material;

and the cost of the production process. When

specific organisms and production technologies
Ch. 5—The Chemical Industry ● 93

Table 1 1.—Some Commercial Enzymes and Their Uses

Enzyme Source Industry and application

Amylase . . . . . . . . . . . . . . . . . . .

Bromelin. . . . . . . . . . . . . . . . . . .

Cellulase and hemicellulase . .

Dextransucrase. . . . . . . . . . . . .

Ficin . . . . . . . . . . . . . . . . . . . . . .

Glucose oxidase (plus catalase

or peroxidase) . . . . . . . . . . . .

Invertase. . . . . . . . . . . . . . . . . . .

Lactase. . . . . . . . . . . . . . . . . . . .

Lipase. . . . . . . . . . . . . . . . . . . . .

Papain . . . . . . . . . . . . . . . . . . . . .

Pectinase . . . . . . . . . . . . . . . . . .

Penicillinase . . . . . . . . . . . . . . .

Pepsin . . . . . . . . . . . . . . . . . . . . .

Protease. . . . . . . . . . . . . . . . . . .

Streptodornase . . . . . . . . . . . . .

Animal(pancreas)

Plant(barley malt)

Fungi (Aspergillus niger,A. oryzae)

Bacteria (Bacillus subtilis)

Plant (pineapple)

Fungi (Aspergillus niger)

Bacteria(Leuconostoc mesenteroides)

Plant (fig latex)

Fungi (Aspergillus niger)

Yeast (Saccharomyces cerevisiae)

Yeast (Saccharomyces fragilis)

Fungi (Aspergillus niger)

Plant(papaya)

Fungi (Aspergillus niger)

Bacteria(Bacillus cereus)

Animal (hog stomach)

Animal (pancreas)

Animal (pepsin)

Animal (rennin, rennet)

Animal (trypsin)

Fungi (Aspergillus oryzae)

Bacteria (Bacillus subtilis)

Bacteria (Streptococcus pyrogenes)

Pharmaceutical digestive aids

Textile: desizing agent

Baking: flour supplement

Brewing, distilling, and industrial alcohol mashing

Food: precooked baby foods

Pharmaceutical digestive aids

Textile: desizing agent

Baking: flour supplement

Brewing, distilling, and industrial alcohol mashing

Food: precooked baby foods, syrup manufacture

Pharmaceutical digestive aids

Paper starch coatings

Starch: cold-swelling laundry starch

Food: meat tenderizer

Pharmaceutical digestive aids

Food: preparation of liquid coffee concentrates

Pharmaceutical preparation of blood-plasma

extenders, and dextran for other uses

Pharmaceutical: debriding agent

Pharmaceutical test paper for diabetes

Food: glucose removal from egg solids

Candy: prevents granulation of sugars in soft-center

candies

Food: artificial honey

Dairy: prevents crystallization of lactose in ice cream

and concentrated milk

Dairy: flavor production in cheese

Brewing: stabilizes chill-proof beer

Food: meat tenderizer

Wine and fruit juice: clarification

Medicine: treatment of allergic reaction to penicillin,

diagnostic agent

Food: animal feed supplement

Dairy: prevents oxidized flavor

Food: protein hydrolysates

Leather: bating

Pharmaceutical: digestive aids

Textile: desizing agent

Brewing: beer stabilizer

Dairy: cheese

Pharmaceutical: wound debridement

Baking: bread

Food: meat tenderizer

Baking: modification of cracker dough

Brewing: clarifier

Pharmaceutical: wound debridement

SOURCE: David Perlman, “The Fermentation Industries,” American Society for Microbiology News 39:10, 1973, p. 653.

have been developed, the cost of raw materials

becomes the limiting step in production. If a

strain of yeast, for example, produces 5 percent

ethanol using sugar as a raw material, the process might become economically competitive if

the cost of sugar drops or the price of petroleum rises. Even if prices remain stable, the

micro-organisms might be genetically improved

to increase their yield; genetic manipulation

might solve the problem of an inefficient

organism. Finally, the production process itself

is a factor. After fermentation, the desired product must be separated from the other compounds in the reaction mixture. As an aid to recovery, the production conditions might be

altered and improved to generate more of a desired compound.

More than one raw material can be used in a

fermentation process. If, in the case of ethanol,
94 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

the price of sucrose (from sugarcane or sugar

beets) is not expected to change, the production

technology is being run at optimum efficiency,

and the micro-organism is producing as much

ethanol as it can, the hurdle to economic competitiveness might be overcome if a less expensive raw material—cellulose, perhaps—were

used. But cellulose cannot be used in its natural

state: physical, chemical, or biological methods

must be devised to break it down to its glucose

(also a sugar) components.

The constraints vary from compound to compound. But even though the role of genetics

must be examined on a product-by-product basis, certain generalizations can be made. Overall, genetic engineering will probably have an

impact on three processes:

●

●

●

Aerobic fermentation, which produces enzymes, vitamins, pesticides, growth regulators, amino acids, nucleic acids, and other

speciality chemicals, is already well-established. Its use should continue to grow. Already, complex biochemical like antibiotics, growth factors, and enzymes are made

by fermentation. Amino acids and nucleotides—somewhat less complicated molecules—are sometimes produced by fermentation. Their production is expected to

increase.

Anaerobic fermentation, which produces

organic acids, methane, and solvents, is the

industry’s area of greatest current growth.

Already, 40 percent of the ethanol manufactured in the United States is produced

in this way. The main constraint on the

production of other organic acids and solvents is the need for cheaper methods for

converting cellulose to fermentable sugars.

Chemical modification of the fermentation

products of both aerobic and anaerobic

fermentation, which to date has rarely

been used on a commercial scale, is of

great interest. (See table 12.) Chemical production technologies that employ high temperatures and pressures might be replaced

by biological technologies operating at atmospheric pressure and ambient temperature. A patent application has already been

filed for the biological production of one of

Table 12.—Expansion of Fermentation Into

the Chemical Industry

Examples

Aerobic fermentation

Enzymes . . . . . . . . . . . . . . . . Amylases, proteases

Vitamins . . . . . . . . . . . . . . . . Riboflavin B,z

Pesticides. . . . . . . . . . . . . . . Bacillus thuringiensis

Growth regulators . . . . . . . . Gibberellin

Amino acids . . . . . . . . . . . . . Glutamic, Iysine

Nucleic acids

Acids. . . . . . . . . . . . . . . . . . . Malic acid, citric acid

Anaerobic fermentation

Solvents . . . . . . . . . . . . . . . . Ethanol, acetone, n-butanol

Acids. . . . . . . . . . . . . . . . . . . Acetic, propionic, acrylic

SOURCE: Office of Technology Assessment.

these products, ethylene glycol, by the

Cetus Corp. in Berkeley, Calif. The process

is claimed to be more energy efficient and

less polluting. If it proves successful when

run at an industrial scale, the technology

could become significant to a [J. S. market

totaling $21/2 billion per year.

The chemical industry produces a variety of

likely targets for biotechnology. Tables I-B-27

through 1-B-32 in appendix I-B present projections of the potential economic impacts of applied genetics on selected compounds that

represent large markets, and the time frames

for potential implementation. Table I-B-7 lists

one large group of organic chemicals that were

identified by the Genex Corp. and Massachusetts Institute of Technology (MIT) as amenable

to biotechnological production methods. They

are in agreement on about 20 percent of the

products cited, which underscores the uncertain nature of attempting to predict so far into

the future.

Fertilizers, polymers, and pesticides

Gaseous ammonia is used to produce nitrogen

fertilizers. About 15 billion tonnes of ammonia

were produced chemically for this purpose, in

1978; the process requires large amounts of

natural gas. Nitrogen can also be converted, or

“fixed,” to ammonia by enzymes in micro-organisms; about 175 billion tonnes are fixed per

year. For example, one square yard of land

planted with certain legumes (such as soybeans)

can fix up to 2 ounces of nitrogen, using bac-
Ch. 5—The Chemical Industry ● 95

teria associated with their roots. Currently, microbial production of ammonia from nitrogen is

not economically competitive. Aside from the

difficulties associated with the enzyme’s sensitivity to oxygen and the near total lack of

understanding of its mechanism, it takes the

equivalent of the energy in 4 kilograms (kg) of

sugar to make 1 kg of ammonia. Since ammonia

costs $0. 13/kg and sugar costs $0.22/kg, it is unlikely that the chemical process will be replaced

in the near future. On the other hand, the genes

for nitrogen fixation have now been transferred

into yeast, opening up the possibility that agriculturally useful nitrogen can be made by fermentation.

A large segment of the chemical industry engaged in the manufacture of polymers is shown

in table 13. A total of 4.3 million tonnes of

fibers, 12 million tonnes of plastics, and I. I million tonnes of synthetic rubber were produced

in the United States in 1978. All were derived

from petroleum, with the exception of the less

than 1 percent derived from cellulose fibers.

The most likely ones are polyamides (chemically

related to proteins), acrylics, isoprene-type rubber, and polystyrene, Because most monomers,

the building blocks of polymers, are chemically

simple and are presently available in high yield

from petroleum, their microbial production in

the next decade is unlikely.

While biotechnoloqy is not ready to replace

the present technology, its eventual impact on

polymer production will probably be large.

Biopolymers represent a new way of thinking.

Most of the important constituents of cells are

polymers: proteins (polypeptides from amino

acid monomers), polysaccharides (from sugar

monomers), and polynucleotides (from nucleotide monomers). Since cells normally assemble

polymers with extreme specificity, the ideal industrial process would imitate the biological

production of polymers in all possible respects—

using a single biological machine to convert a

raw material, e.g., a sugar, into the monomer to

polymerize it, then to form the final product. A

more likely application is the development of

new monomers for specialized applications.

Polymer chemistry has largely consisted of the

study of how their properties can be modified.

Table 13.—The Potential of Some Major Polymeric

Materials for Production Using Biotechnology

Domestic production 1978

Product (thousand tonnes)

Plastics

Thermosetting resins

Epoxy. . . . . . . . . . . . . . . . . . . . . . . . .

Polyester. . . . . . . . . . . . . . . . . . . . . .

Urea. . . . . . . . . . . . . . . . . . . . . . . . . .

Melamine . . . . . . . . . . . . . . . . . . . . .

Phenolic . . . . . . . . . . . . . . . . . . . . . .

Thermoplastic resins

Polyethylene

Low density . . . . . . . . . . . . . . . . .

High density. . . . . . . . . . . . . . . . .

Polypropylene . . . . . . . . . . . . . . . . .

Polystyrene. . . . . . . . . . . . . . . . . . . .

Polyamide, nylon type. . . . . . . . . . .

Polyvinyl alcohol . . . . . . . . . . . . . . .

Polyvinyl chloride. . . . . . . . . . . . . . .

Other vinyl resins. . . . . . . . . . . . . . .

Fibers

Cellulosic fibers

Acetate . . . . . . . . . . . . . . . . . . . . . . .

Rayon . . . . . . . . . . . . . . . . . . . . . . . .

Noncellulosic fibers

Acrylic. . . . . . . . . . . . . . . . . . . . . . . .

Nylon. . . . . . . . . . . . . . . . . . . . . . . . .

Olefins. . . . . . . . . . . . . . . . . . . . . . . .

Polyester. . . . . . . . . . . . . . . . . . . . . .

Textile glass . . . . . . . . . . . . . . . . . . .

Other . . . . . . . . . . . . . . . . . . . . . . . . .

Rubbers

Styrene-butadiene. . . . . . . . . . . . . . . .

Polybutadiene . . . . . . . . . . . . . . . . . . .

Butyl . . . . . . . . . . . . . . . . . . . . . . . . . . .

Nitrile. . . . . . . . . . . . . . . . . . . . . . . . . . .

Polychlorophene . . . . . . . . . . . . . . . . .

Ethylene-propylene . . . . . . . . . . . . . . .

Polyisoprene. . . . . . . . . . . . . . . . . . . . .

135

544

504

90

727

3,200

1,890

1,380

2,680

124

57

2,575

88

139

269

327

1,148

311

1,710

418

7

628

170

69

33

72

78

62

SOURCE: Office of Technology Assessment.

Conceivably, biotechnology could enable the

modification of their function and form.

Pesticides include fungicides, herbicides, insecticides, rodenticides, and related products

such as plant growth regulators, seed disinfectants, soil conditioners, and soil fumigants. The

largest market (roughly $500 million annually)

involves the chemical and microbial control of

insects. Although microbial insecticides have

been around for years, they comprise only 5

percent of the market. However, recent successes in developing viruses and bacteria that

produce diseases in insects, and the negative

publicity given to chemical insecticides, have

encouraged the use of microbial insecticides.
96 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

The other two species can be produced by conVentional fermentation techniques. They have

been useful because they form spores that can

be mass-produced easily and are stable enough

to be handled commercially. The actual substances that cause toxicity to the insect are toxins synthesized by the microbes.

Genetic engineering should make it possible

to construct more potent bacterial insecticides

by increasing the dosage of the genes that code

for the synthesis of the toxins involved. Mixtures of genes capable of directing the synthesis

of various toxins might also be produced.

Constraints on biological production techniques ____

The chief impediments to using biological

production technology are associated with the

need for biomass. 2 They include:

●

●

●

●

●

●

●

competition with food needs for starch and

sugar;

cyclic availability;

biodegradability and associated storage

problems;

high moisture content for cellulosics, and

high collection and storage costs;

mechanical processing for cellulosics;

the heterogeneous nature of cellulosics (mixtures of cellulose, hemicellulose, and lignin); and

The need for disposal of the nonfermentable port ions of the biomass.

For food-related biomass sources, such as sugar, corn, and sorghum, few technological barriers exist for conversion to fermentable sugars;

but subsidies are needed to make the fermentation of sugars as profitable as their use as food.

For cellulosic biomass sources such as agricultural wastes, municipal wastes, and wood, technological barriers exist in collection, storage,

pretreatment, fermentation, and waste disposal.

In addition, biomass must always be transformed into sugars by either chemical or enzymatic processes before fermentation can

begin.

‘ljnf~r,qv F’r(ml ~io/(J,gif’if/ PrOct’s.st*,s, 01). (’it.

A second major impediment is associated

with the purification stage of production. Most

chemical products of fermentation are present

in extremely dilute solutions, and concentrating

these solutions to recover the desired product is

highly energy-intensive. Problems of technology

and cost will continue to make this stage an important one to improve.

The developments in genetics show great

promise for creating more versatile micro-organisms, but they do not by themselves produce a

cheaper fuel or plastic. Associated technologies

still require more efficient fermentation facilities and product separation processes; microbes may produce molecules, but they will

not isolate, purify, concentrate, mix, or package

them for human use.

The interaction between genetic engineering

and other technologies is illustrated by the

problems of producing ethanol by fermentation. The case study presented in appendix I-D

identifies those steps in the biomass-to-ethanol

scheme that need technological improvements

before the process can become economical.

Genetic engineering is expected to reduce

costs in many production steps. For certain

ones—such as the pretreatment of the biomass

to make it fermentable—genetics will probably

not play a role: physical and chemical technologies will be responsible for the greatest advances. For others, such as distillation, genetic
Ch. 5—The Chemical Industry . 97

technologies should make it possible to engineer

organisms that can ferment at high temperatures (82° to 85° C) so that the fermentation and

at least part of the distillation can both take

place in the same reactor. Various technol-

“’1’11(~1.lllfjl]llilit (’tllill}ol 1)1’()(111(’(’1’S llil\’f’ illl’(’il(l~r 1)[’[’11 IIescrikd

ill IIW g(~tlus [,’l(jslri(fiunl [i. (’., (;. fhf’rr?~()(:f;ll[~n~). Ill i![ldil ion, gciwl i-

(’ill 11’ wlgilwf’1’(~d ol’~illlisllls, (Ies(’ril)e(i ii!+ il (’1’OSS 1)(’ltt’tWll V(!ilSIS

;111( i I I)ol’lllol)ll” i] it’ I)il(’1 f>l’lil, (’iIll I’pl’lllf)tll il I 700 (:.

An overview of impacts

The cost of raw materials may become cheaper than the petroleum now used—especially if

cellulose conversion technologies can be developed. The source of raw materials would also

be broader since several kinds of biomass could

be interchanged, if necessary. For small quantities of chemicals, the raw material supply

would be more dependable, particularly because of the domestic supply of available biomass. For substances produced in large quantities, such as ethanol, the supply of biomass

could limit the usefulness of biotechnology.

Raw materials, such as organic wastes, could

be processed both to produce products and

reduce pollution. Nevertheless, the impact on

total imported petroleum will be low. Estimates

of the current consumption of petroleum as a

raw material for industrial chemicals is approximately 5 to 8 percent of the total imported.

Impacts on the process include relatively

cheaper production costs for selected compounds. For these, lower temperatures and

pressures can be used, suggesting that the processes might be safer. Chemical pollution from

biotechnology may be lower, although methods

of disposal or new uses must be found for the

micro-organisms used in fermentation. Finally,

the biological processes will demand the development of new technologies for the separation

and purification of the products.

impacts on the products include both cheaper

existing chemicals as well as entirely new products. Since biotechnology is the method of

choice for producing enzymes, new uses for enogies, such as the immobilization of whole cells

in reactor columns, could be developed in parallel with genetic technologies to increase the stability of cells in fermenters.

‘l”lIt? ild\’illltit#?S Ot” SllCh Ilwrnmphilic I“t’l’lll(’lllilt iollS ill’t’ signif’i-

(’illll: I’t?l’lllt%ltiitioll I inw is CollSi&Will)l}” l’t’(111(’tXl; IIIP risk ot (’ollIillllillill ion is llt?ill’1.V f?litllitlille(l: illl(l cooling” t.t’(lltil.(’lllt’tlls ill’(’

I[)w’t?t’ (111(+ 10 llle lli@el. lt?lll~t?l’iitll l’(’ ot” III(’ t’msnlt’nt in~ 1)1’0111.

zymes may expand and drive this sector of the

industry.

Impacts on other industries

Although genetic engineering will develop

new techniques for synthesizing many substances, the direct displacement of any present

industry appears to be doubtful: Genetic engineering should be considered simply another industrial tool. As such, any industry’s response

should be to use this technique to maintain its

positions in its respective markets. The point is

illustrated by the variety of companies in the

pharmaceutical, chemical, and energy industries that have invested in or contracted with

genetic engineering firms. Some large companies are already developing inhouse genetic

engineering research capabilities.

The frequent, popular reference to the small,

innovative “genetic engineering companies” as a

major new industry is somewhat misleading.

The companies (see table 14) arose primarily to

convert micro-organisms with little commercial

use into micro-organisms with commercial potential. A company such as the Cetus Corp. initially used mutation and selection to improve

strains, whereas other pioneers such as Genentech, Inc., Biogen, S. A., and Genex Corp. were

founded to exploit recombinant DNA (rDNA)

technology. Part of their marketing strategy includes the sale or licensing of genetically engineered organisms to large established commercial producers in the chemical, pharmaceutical,

food, energy, and mining industries. Each engi-
98 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Table 14.—Some Private Companies With Biotechnology Programs

Approximate Research capacity

Company Founded employees 1979 Ph. D.s 1979 Recombinant DNA Hybridomas

Atlantic Antibodies. . . . . . . . . . . . . . . . . . 1973 50 2 x

Bethesda Research Laboratories . . . . . . 1976 130 x x

Biogen . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1978 30 (50b

) (18

b

)

(3)(5)

x x

Bens Bio Logicals . . . . . . . . . . . . . . . . . . . 1979 15 10 x x

Centocor. . . . . . . . . . . . . . . . . . . . . . . . . . . 1979 20(1)-10(4) x

Cetus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1972 250 50 x x

Clonal Research . . . . . . . . . . . . . . . . . . . . 1979 6 1 x

Collaborative ResearchC

. . . . . . . . . . . . . . 1961 85 15 x x

(Collaborative Genetics) . . . . . . . . . . . . (1979) (4) (3) x

Genentech . . . . . . . . . . . . . . . . . . . . . . . . . 1976 90 30 x

Genex . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1977 30 12 x

Hybritech . . . . . . . . . . . . . . . . . . . . . . . . . . 1978 3 3

(1) 6 x

Molecular Genetics. . . . . . . . . . . . . . . . . . 1979 6(4) x x

Monoclinal Antibodies. . . . . . . . . . . . . . . 1979 6 x

NewEngland Biolabs . . . . . . . . . . . . . . . . 1974 22(22)-5 (4) x

q=.E~r9t~t~co. estimates.

bEXwtedby~cem~rl~.

ccollaboratlve Research isamajorowner of Collaborative Genetics. Thedivision batweenthem isnotyetdistinct.

SOURCES: (l)Sc/ence206, p.692.693, 1960(52 peopletoexpandto100 by1961).

(2) Science 206, p. 692-693, 19S0 (20 senior peraona).

(3) Science 206, p. 692-693, 1960(16 scientists, 30 employees).

(4) Dun & Bradstreet, Inc.

(5) Chemlcalarrd Engineering News, Mar. 19,1960.

Office of Technology Assessment.

neering firm also intends to manufacture some

products itself. It is likely that the products reserved for inhouse manufacture will be lowvolume, high-priced compounds like interferon.

Genetic engineering by itself is a relatively

small-scale laboratory operation. Consequently,

genetic engineering firms will continue to offer

services to companies that do not intend to

develop this capacity in their own inhouse laboratories. Specifically, a genetic engineering

company may contract with a firm to develop a

biological production method for its products.

At the same time, larger companies might establish inhouse staffs to develop biological methods

for both old and new products. (Several larger

companies already have more inhouse genetic

engineering personnel than some of the independent genetic engineering companies.)

In addition, suppliers of genetic raw materials

may decide to expand into the production of

genetically engineered organisms. Suppliers of

restriction endonuclease enzymes for example,

which are used in constructing rDNA, have

already entered the field. Diagnostic firms could

develop new bioassays for which they themselves would guarantee a market. Finally, companies with byproducts or waste products are

beginning to examine the possibility of converting them into useful products. This approach

(which is somewhat more developed in Europe)

assumes that with the proper technology the

waste materials can become a resource.

Some industries, including manufacturers of

agitators (drives), centrifuges, evaporators, fermenters, dryers, storage tanks and process

vessels, and control and instrumentation systems, might profit by producing equipment

associated with fermentation.

Impacts on university research

From the beginning, genetic engineering

firms established strong ties with universities.

These were responsible for providing most of

the scientific knowledge that formed the basis

for applied genetics as well as the initial scientific workforce:

Cetus Corp. established a pattern by recruiting a prestigious Board of Scientific

Advisors who remain in academic positions.

Genentech, Inc., cofounded by a professor

at the University of California at San Fran-
Ch. 5—The Chemical Industry . 99

●

●

●

cisco, initially depended largely on outside

scientists.

Biogen, S. A., was organized by professors

at Harvard and MIT plus six European scientists, and placed R&D contracts with academic researchers.

Collaborative Genetics has a Nobel prize

winner from MIT as the chairman of its scientific advisory board.

Hybritech, Inc., has as its scientific nucleus

a University of California, San Diego, professor complemented by scientists at the

Salk Institute.

In addition to these companies, others have also

been establishing closer ties with the academic

community.

Much of the research that will be useful to industry will continue to be carried out in university laboratories. At present, it is often difficult

to decide whether a research project should be

classified as “basic” (generally more interesting

to an academic researcher) or “applied” (generally more interesting to industry). E.g., a change

in the genetic code, which increases gene activity, would be just as exciting to a basic scientist

as to an industrial one.

This dialog between the universities and industry—both through formal and informal arrangements –has fostered innovation. Although

the number of patents applied for is not a direct

reflection of the level of innovation, it is still one

indication. By the end of 1980, several hundred

patent applications were filed for genetically

engineered micro-organisms, their products,

and their processes.

University research has clearly affected industrial development, and has in turn been affected by industry. Although the benefits are

easily recognized, some drawbacks have been

suggested. The most serious is the concern that

university scientists will be restrained in their

academic pursuits and in their exchange of information and research material. To date, anecdotal information suggests that some scientists

are being more circumspect about sharing information. Still, secrecy is not new to highly

competitive areas of biomedical research. In addition, scientists in other academic disciplines

useful to industry-such as chemistry and physics—have managed to achieve a balance between secrecy and openness.

The social impacts of local

industrial activity

Despite the extensive media coverage of

rDNA and other forms of genetic engineering,

there is little evidence that people who live near

companies using such techniques are still greatly concerned about possible hazards. This may

be partly owing to a lack of awareness that a

particular company is doing genetic research

and partly because companies thus far have

adhered to the National institutes of Health

(NIH) Guidelines. Some companies have placed

individuals on their institutional biosafety committees who are respected and trusted members of the local community. By involving the

local citizens with no vested corporate interest,

a mechanism for oversight has been provided.

(For a more detailed discussion, see ch. 11.)

Impacts on manpower

Two types of impacts on workers can be expected:

● The creation of jobs that replace those held

by others. E.g., a worker involved in

chemical production might be replaced by

one producing the same product biologically.

● The creation of new jobs.

Workers in three categories would be affected:

● those actually involved in the fermentationproduction phase of the industry;

● those involved in the R&D phase of the industry, particularly professionals; and

. those in support industries.

Projections of manpower requirements are

only as accurate as the projections of the level of

industrial activity. In the past 5 years, about 750

new jobs have been created within the small genetic engineering firms (including monoclinal

antibody producers). Of these, approximately

one-third hold Ph. D. degrees.
100 . Impacts of Applied Genetics—Micro-0rganisms, Plants, and Animals

Data obtained through an (OTA survey of 284

firms indicate that the pharmaceutical industry

employs the major share of personnel working

in applied genetics programs. (See table 15. ) The

average number of Ph. D.s in each industry is

given in table 16. A rough estimate of professional scientific manpower at this level includes:

6 in food, 45 in chemical, 120 in pharmaceutical,

and 18 in specialty chemicals-a total of 189. If

the number of research support personnel is

approximately twice the number of Ph. D.s, the

total rises to about 570. If $165,000 per year is

required to support one Ph. D. in industry, the

total value of such manpower is approximately

$31 million.

Estimates of the number of companies engaged in applied genetics work in 1980 can be

compared with the total number of firms with

fermentation activities. A tabulation of firms on

a worldwide basis in 1977 revealed 145 companies, of which 27 were American. (See table

17. ) These companies produced antibiotics, enzymes, solvents, vitamins and growth factors,

Table 15.—Distribution of Applied Genetics

Activity in Industry

Distribution of applied

genetics activity by Percent

Classification company classaof total

Food . . . . . . . . . . . . . . (6146) 13

Chemical . . . . . . . . . . (9/52) 17

Pharmaceutical . . . . . (12/25) 48

Specialty chemicalb . (6/68) 9

algnoreg small firing specializing in genetic research.

bFood ingredients, reagents, enzymes.

SOURCE: Office of Technology Assessment.

Table 16.—Manpower (Iow.(average)-high) Distribution

of a Firm With Applied Genetics Activity

Ph. D. M.S. Bachelors

Food. . . . . . . . . . . . . . . . . ql)-2

Chemical . . . . . . . . . . . . . 3-(5)-7

Pharmaceutical. . . . . . . 2-(10)-24

Specialty. . . . . . . . . . . . . l-(3)-8

Biotechnology

Genetic engineering. 3-(15)-32

Hybridoma. . . . . . . . . . l-(3)-6

Other . . . . . . . . . . . . . . O-(2)-4

Average . . . . . . . . . .1-(6)-12

0-(1 )-2

0-(1 )-2

l-(4)-9

l-(3)-4

2-(1 1)-20

0-(2)-0

2-(4)-6

l-(4)-6

O-(2)-8

2-(5)-7

1-(8)-20

2-(2)-4

5-(15)-25

0-(20)-0

8-(10)-13

3-(8)-12

SOURCE: Office of Technology Assessment.

Table 17.—lndex to Fermentation Companies

1. Abbott Laboratories, North Chicago, 111.

2. American Cyanamid, Wayne, N.J.

3. Anheuser-Busch, Inc., St. Louis, Mo.

4. Bristol-Myers Co., Syracuse, N.Y.

5. Clinton Corn Processing Co., Clinton, Iowa

6. CPC International, Inc., Argo, Ill.

7. Dairyland Laboratories, Inc., Waukesha, Wis.

8. Dawe’s Laboratories, Inc., Chicago Heights, Ill.

9. Grain Processing Corp., Muscatine, Iowa

10. Hoffman-LaRoche, Inc., Nutley, N.J.

11. IMC Chemical Group, Inc., Terre Haute, Ind.

12. Eli Lilly & Co., Indianapolis, Ind.

13. Merck & Co., Inc., Rahway, N.J.

14. Miles Laboratories, Inc., Elkhart, Ind.

15. Parke, Davis & Co., Detroit, Mich.

16. S. B. Penick & Co., Lyndhurst, N.J.

17. Pfizer, Inc., New York, N.Y.

18. Premier Malt Products, Inc., Milwaukee, Wis.

19. Rachelle Laboratories, Inc., Long Beach, Cal if.

20. Rohm & Haas, Philadelphia, Pa.

21. Schering Corp., Bloomfield, N.J.

22. G. D. Searle & Co., Skokie, Ill.

23. E. R. Squibb& Sons, Inc., Princeton, N.J.

24. Standard Brands, Inc., New York, N.Y.

25. Stauffer Chemical Co., Westport, Corm.

26. Universal Foods Corp., Milwaukee, Wis.

27. The Upjohn Co., Kalamazoo, Mich.

28. Wallerstein Laboratories, Inc., Morton Grove, Ill.

29. Wyeth Laboratories, Philadelphia, Pa.

SOURCE: Office of Technology Assessment.

nucleosides, amino acids, and miscellaneous

products. (See table 18.) The only chemical firm

listed was the Stauffer Chemical Co. Ten firms

are listed as having the ability to produce food

and feed yeast. (See table 19. ) Correcting for

firms listed twice, at least 38 U.S. firms were

engaged in significant fermentation activity for

commercial products, excluding alcoholic beverages, in 1977. Not all have research expertise

in fermentation or biotechnology, much less a

regular genetics program: 10 to 20 were in the

chemical industry; 25 to 40 in fermentation (enzyme, pharmaceutical, food, and specialized

chemicals); and 10 to 15 in biotechnology (genetic engineering)—or about 45 to 75 firms in all.

If average manpower numbers are used, the

total number of professionals involved in commercial applied genetics research is:

Ph. D.s: 300-450

Others: 600-900

900-1,350

The number of workers that will be involved

in the production phase of biotechnology repre-
Ch.5—The Chemical Industry ● 101

Table 18.-Fermentation Products and Producers

Product Some producers Product Some producers

Amino acids

L-alanine. . . . . . . . . . . . . . . . . . . . . . . . . . .

L-arginine. . . . . . . . . . . . . . . . . . . . . . . . . .

L-aspartic acid. . . . . . . . . . . . . . . . . . . . . .

L-citrulline . . . . . . . . . . . . . . . . . . . . . . . . .

L-glutarnic acid . . . . . . . . . . . . . . . . . . . . .

L-glutamine . . . . . . . . . . . . . . . . . . . . . . . .

L-glutathione . . . . . . . . . . . . . . . . . . . . . . .

L-histidine . . . . . . . . . . . . . . . . . . . . . . . . .

L-homoserine. . . . . . . . . . . . . . . . . . . . . . .

L-isoleucine . . . . . . . . . . . . . . . . . . . . . . . .

L-leucine. . . . . . . . . . . . . . . . . . . . . . . . . . .

L-lysine . . . . . . . . . . . . . . . . . . . . . . . . . . . .

L-methionine . . . . . . . . . . . . . . . . . . . . . . .

L-ornithine . . . . . . . . . . . . . . . . . . . . . . . . .

L-phenylalanine. . . . . . . . . . . . . . . . . . . . .

L-proline. .... . . . . . . . . . . . . . . . . . . . . .

L-serine. . . . . . . . . . . . . . . . . . . . . . . . . . . .

L-threonine. .. . . . . . . . . . . . . . . . . . . . . .

L-tryptophan. . . . . . . . . . . . . . . . . . . . . . .

L-tyrosine . . . . . . . . . . . . . . . . . . . . . . . . . .

L-valine. ..... . . . . . . . . . . . . . . . . . . . . .

Miscellaneous products and processes

Acetoin . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Acyloin . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Anka-pigment(red) . . . . . . . . . . . . . . . . . .

Blue cheese flavor. . . . . . . . . . . . . . . . . . .

Desferrioxamine . . . . . . . . . . . . . . . . . . . .

Dihydroxyacetone . . . . . . . . . . . . . . . . . . .

Dextran . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Diacetyl (from acetoin) . . . . . . . . . . . . . . .

Ergocornine . . . . . . . . . . . . . . . . . . . . . . . .

Ergocristine . . . . . . . . . . . . . . . . . . . . . . . .

Ergocryptine . . . . . . . . . . . . . . . . . . . . . . .

Ergometrine . . . . . . . . . . . . . . . . . . . . . . . .

Ergotamine. . . . . . . . . . . . . . . . . . . . . . . . .

Bacillus thuringiensis insecticide. . . . . .

Lysergic acid . . . . . . . . . . . . . . . . . . . . . . .

Paspalic acid . . . . . . . . . . . . . . . . . . . . . . .

Picibanil . . . . . . . . . . . . . . . . . . . . . . . . . . .

Ribose. . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Scteroglucan . . . . . . . . . . . . . . . . . . . . . . .

Sorbose(from sorbitol). . . . . . . . . . . . . . .

Starter cultures . . . . . . . . . . . . . . . . . . . . .

Sterol oxidations.. . . . . . . . . . . . . . . . . . .

Steroid oxidations. . . . . . . . . . . . . . . . . . .

Xanthan . . . . . . . . . . . . . . . . . . . . . . . . . . .

Antibiotics

Adriamycin. . . . . . . . . . . . . . . . . . . . . . . . .

Amphomycin . . . . . . . . . . . . . . . . . . . . . . .

Amphotericin B.. . . . . . . . . . . . . . . . . . . .

Avoparcin . . . . . . . . . . . . . . . . . . . . . . . . . .

Azalomycin F . . . . . . . . . . . . . . . . . . . . . . .

Bacitracin. . . . . . . . . . . . . . . . . . . . . . . . . .

Bambermycins. . . . . . . . . . . . . . . . . . . . . .

Bicyclomycin. . . . . . . . . . . . . . . . . . . . . . .

Blasticidin S. . . . . . . . . . . . . . . . . . . . . . . .

Bleomycin . . . . . . . . . . . . . . . . . . . . . . . . .

Cactinomycin

Candicidin B. . . . . . . . . . . . . . . . . . . . . . . .

Candidin . . . . . . . . . . . . . . . . . . . . . . . . . . .

25

13

7

17,21,28

1

10,17

7,13,14

22,27

21,23,27,29

13,17

23

2

11,16,17

16

Capreomycin . . . . . . . . . . . . . . . . . . . . . . .

Cephalosporins. . . . . . . . . . . . . . . . . . . . .

Chromomycin A3 . . . . . . . . . . . . . . . . . . . .

Colistin . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Cycloheximide. . . . . . . . . . . . . . . . . . . . . .

Cycloserine . . . . . . . . . . . . . . . . . . . . . . . .

Dactinomycin. . . . . . . . . . . . . . . . . . . . . . .

Daunorubicin. . . . . . . . . . . . . . . . . . . . . . .

Destomycin . . . . . . . . . . . . . . . . . . . . . . . .

Enduracidin . . . . . . . . . . . . . . . . . . . . . . . .

Erythromycin . . . . . . . . . . . . . . . . . . . . . . .

Fortimicins. . . . . . . . . . . . . . . . . . . . . . . . .

Fumagiliin . . . . . . . . . . . . . . . . . . . . . . . . .

Fungimycin . . . . . . . . . . . . . . . . . . . . . . . .

Fusidic acid . . . . . . . . . . . . . . . . . . . . . . . .

Gentamicins. . . . . . . . . . . . . . . . . . . . . . . .

Gramicidin A . . . . . . . . . . . . . . . . . . . . . . .

Gramicidin J(S) . . . . . . . . . . . . . . . . . . . . .

Griseofulvin . . . . . . . . . . . . . . . . . . . . . . . .

Hygromycin B . . . . . . . . . . . . . . . . . . . . . .

Josamycin . . . . . . . . . . . . . . . . . . . . . . . . .

Kanamycins. . . . . . . . . . . . . . . . . . . . . . . .

Kasugamycin. . . . . . . . . . . . . . . . . . . . . . .

Kitasatamycin. . . . . . . . . . . . . . . . . . . . . .

Lasalocid . . . . . . . . . . . . . . . . . . . . . . . . . .

Lincomycin. . . . . . . . . . . . . . . . . . . . . . . . .

Lividomycin . . . . . . . . . . . . . . . . . . . . . . . .

Macarbomycin. . . . . . . . . . . . . . . . . . . . . .

Mepartricin. . . . . . . . . . . . . . . . . . . . . . . . .

Midecamycin. . . . . . . . . . . . . . . . . . . . . . .

Mikamycins . . . . . . . . . . . . . . . . . . . . . . . .

Mithramycin. . . . . . . . . . . . . . . . . . . . . . . .

Mitomycin C. . . . . . . . . . . . . . . . . . . . . . . .

Mocimycin . . . . . . . . . . . . . . . . . . . . . . . . .

Monensin . . . . . . . . . . . . . . . . . . . . . . . . . .

Myxin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Neornycins. ... . . . . . . . . . . . . . . . . . . . .

Novobiocin. . . . . . . . . . . . . . . . . . . . . . . . .

Nystatin . . . . . . . . . . . . . . . . . . . . . . . . . . .

Oleandomycin. . . . . . . . . . . . . . . . . . . . . .

Oligomycin. . . . . . . . . . . . . . . . . . . . . . . . .

Paromomycins. . . . . . . . . . . . . . . . . . . . . .

Penicillin G. . . . . . . . . . . . . . . . . . . . . . . . .

Penicillin V . . . . . . . . . . . . . . . . . . . . . . . . .

Penicillins(semisynthetic). . . . . . . . . . . .

Pentamycin . . . . . . . . . . . . . . . . . . . . . . . .

Pimaricin . . . . . . . . . . . . . . . . . . . . . . . . . .

Polymyxins. . . . . . . . . . . . . . . . . . . . . . . . .

Polyoxins . . . . . . . . . . . . . . . . . . . . . . . . . .

Pristinamycins. . . . . . . . . . . . . . . . . . . . . .

Quebemycin. . . . . . . . . . . . . . . . . . . . . . . .

Ribostamycin. . . . . . . . . . . . . . . . . . . . . . .

Rifamycins. . . . . . . . . . . . . . . . . . . . . . . . .

Sagamicin. . . . . . . . . . . . . . . . . . . . . . . . . .

Salinomycin, . . . . . . . . . . . . . . . . . . . . . . .

Siccanin . . . . . . . . . . . . . . . . . . . . . . . . . . .

Siomycin. . . . . . . . . . . . . . . . . . . . . . . . . . .

Sisomicin ... . . . . . . . . . . . . . . . . . . . .

Spectinomycin. . . . . . . . . . . . . . . . . . . . . .

Streptomycin. . . . . . . . . . . . . . . . . . . . . .

Tetracyclines

Clortetracycline. . . . . . . . . . . . . . . . . . . . .

4,12

27

11

13

17,27

21

28

12

4

10

27

17

4

10

16,17,23,27

27

23

17

15

4,12,13,17,23,29

1,4,12,17,23,29

4,13,17,23,29

17

21

27

13,17,29

19

76-565 0 - 81 - 8
102 . Impacts of Applied Genetics—Micro-Organkms, Plants, and Animals

Table 18.-Fermentation Products and Producers

Product Some producers Product Some producers

Demeclocycline. . . . . . . . . . . . . . . . . . . . .

Oxytetracycline. . . . . . . . . . . . . . . . . . . . .

Tetracycline. . . . . . . . . . . . . . . . . . . . . . . .

Tetranactin. . . . . . . . . . . . . . . . . . . . . . . . .

Thiopeptin . . . . . . . . . . . . . . . . . . . . . . . . .

Thiostrepton . . . . . . . . . . . . . . . . . . . . . . .

Tobramycin . . . . . . . . . . . . . . . . . . . . . . . .

Trichomycin. . . . . . . . . . . . . . . . . . . . . . . .

Tylosin . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Tyrothricin ..... . . . . . . . . . . . . . . . . . . .

Tyrocidine . . . . . . . . . . . . . . . . . . . . . . . . .

Uromycin . . . . . . . . . . . . . . . . . . . . . . . . . .

Vaildamycin . . . . . . . . . . . . . . . . . . . . . . . .

Vancomycin . . . . . . . . . . . . . . . . . . . . . . . .

Variotin. . . . . . . . . . . . . . . . . . . . . . . . . . . .

Viomycin. .. ..... . . . . . . . . . . . . . . . . .

Virginiamycin. . . . . . . . . . . . . . . . . . . . . .

Enzymes

Amylases . . . . . . . . . . . . . . . . . . . . . . . . . .

Amyloglucosidase. . . . . . . . . . . . . . . . . . .

Anticyanase. . . . . . . . . . . . . . . . . . . . . . . .

L-asparaginase . . . . . . . . . . . . . . . . . . . . .

Catalase . . . . . . . . . . . . . . . . . . . . . . . . . . .

Cellulase. . . . . . . . . . . . . . . . . . . . . . . . . . .

Dextranase. . . . . . . . . . . . . . . . . . . . . . . . .

‘Diagnostic enzymes’ . . . . . . . . . . . . . . . .

Esterase-lipase . . . . . . . . . . . . . . . . . . . . .

Glucanase . . . . . . . . . . . . . . . . . . . . . . . . .

Glucose dehydrogenase. . . . . . . . . . . . . .

Glucose isomerase . . . . . . . . . . . . . . . . . .

Glucose oxidase . . . . . . . . . . . . . . . . . . . .

Glutamic decarboxylase. . . . . . . . . . . . . .

Hemi-celiulase. . . . . . . . . . . . . . . . . . . . . .

Hespiriginase. . . . . . . . . . . . . . . . . . . . . . .

lnvertase. . . . . . . . . . . . . . . . . . . . . . . . . . .

2

17,19

2,4,17,19,23,27

23

12

12

16,28

12

5,19,20,24,28

5,6,14,28

8,14

6,20,28

28

28

3,5,14,24

8,14

18

14,20,28

24,26,28

Lactase . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Lipase . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Microbial rennet. . . . . . . . . . . . . . . . . . . . .

Naringinase . . . . . . . . . . . . . . . . . . . . . . . .

Pectinase . . . . . . . . . . . . . . . . . . . . . . . . . .

Pentosanase . . . . . . . . . . . . . . . . . . . . . . .

Proteases . . . . . . . . . . . . . . . . . . . . . . . . . .

Streptokinase-streptodornase. . . . . . . . .

Uricase . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Organic acids

Citric acid . . . . . . . . . . . . . . . . . . . . . . . . . .

Comenic acid. . . . . . . . . . . . . . . . . . . . . . .

Erythorbic acid. . . . . . . . . . . . . . . . . . . . . .

Gluconic acid. . . . . . . . . . . . . . . . . . . . . . .

Itaconic acid. . . . . . . . . . . . . . . . . . . . . . . .

2-keto-D-giuconic acid . . . . . . . . . . . . . . .

~-ketoglutaric acid. . . . . . . . . . . . . . . . . . .

Lactic acid . . . . . . . . . . . . . . . . . . . . . . . . .

Malic acid . . . . . . . . . . . . . . . . . . . . . . . . . .

Urocanic acid. . . . . . . . . . . . . . . . . . . . . . .

Solvents

Ethanol . . . . . . . . . . . . . . . . . . . . . . . . . . . .

2,3-butanediol . . . . . . . . . . . . . . . . . . . . . .

Vitamins and growth factors

Gibbereliins . . . . . . . . . . . . . . . . . . . . . . . .

Riboflavin . . . . . . . . . . . . . . . . . . . . . . . . . .

Vitamin B12. . . . . . . . . . . . . . . . . . . . . . . . .

Zearalanol. . . . . . . . . . . . . . . . . . . . . . . . . .

Nucleosides and nucleotides

5-ribonucieotides and nucleosides . . . . .

Orotic acid . . . . . . . . . . . . . . . . . . . . . . . . .

Ara-A-(9-B-D-arabino-furanosyl) . . . . . . ..

6-azauridine . . . . . . . . . . . . . . . . . . . . . . . .

28

20

17,28

28

20,28

20,28

14,17,18,20,28

2

14,17

17

4,17,18

17

17

5

9

1,12,13

13

13

11

15

aBlankm=n~ noU.S.pr~ucer in 1977;therefore, l~produeedbyoneormore foreign flrms(fromatleast 120different firms).

SOURCE: OtticeotTechnology Assessment.

sents a major impact of genetic engineering. To

estimate this number these two calculations

must be made:

● the value or volume of chemicals that

might be produced by fermentation, and

● the number of production workers needed

per unit volume of chemicals produced.

Any prediction of the potential volume of

chemicals is necessarily filled with uncertainties. The approximate market value of organic

chemicals produced in the United States is given

in appendix I-B. Total U.S. sales in 1979 were

calculated to be over $42 billion. On the basis of

the assumptions made, $522 million worth of

bulk organic chemicals could be commercially

produced by genetically engineered strains in

10 years and $7.1 billion in 20 years. Table

I-B-l0 in appendix I-B lists the potential markets

for pharmaceuticals. Excluding methane production, the total potential market for products

obtained from genetically engineered organisms is approximately $14.6 billion.

If the production of chemicals having this

value is carried out by fermentation, it impossible to calculate how many workers will be

needed. Data obtained from industrial sources

reveal that 2 to 5 workers, including those in

supervision, services, and production, are required for $l million worth of product. Hence,

30,000 to 75,000 workers would be required for

the estimated $14.6 billion market.
Ch. 5—The Chemical lndustry ● 103

Table 19.—U.S. Fermentation Companies

Producers of Baker’s yeast and food/feed yeast in

the United States in 1977

Baker’s yeast:

American Yeast Co., Baltimore, Md.

Anheuser-Busch, Inc., St. Louis, Mo.

Federal Yeast Co. (now Diamond Shamrock),

Baltimore, Md.

Fleischmann Yeast Co., New York, N.Y.

Universal Foods Corp., Milwaukee, Wis.

Food/feed yeast:

Amber Laboratories, Juneau, Wis.

Amoco Foods Co., Chicago, Ill.

Boise-Cascade, Inc., Portland, Oreg.

Diamond Mills, Inc., Cedar Rapids, Iowa

Fleischmann Yeast Co., New York, N.Y.

Lakes States Yeast Co., Rhinelander, Wis.

Stauffer Chemical Co., Westport, Corm.

Enzyme producers, 1977

Clinton Corn Processing Co., Clinton, lowa

Miles Laboratories, Inc., Elkhart, Ind.

Premier Malt Products, Inc., Milwaukee, Wis.

SOURCE: Compiled by Perlman, Arner/can Soc/ety for M/crobio/ogy News 43:2,

1977, pp 82-89.

Since the chemicals considered above are

currently being produced, any new jobs in biotechnology will displace the old ones in the

chemical industry. Whether the change will result in a net loss or gain in the number of jobs is

difficult to predict. However, a rough estimate

indicates that approximately the same number

of workers will be required per unit of output.

Estimates of the number of workers are divided into: 1) workers directly involved in the

growth of the organisms; and 2) workers involved in the "recovery" phase, where the

organisms are harvested and the chemical product is extracted, purified, and packaged. Based

on industry data, the number of workers in the

fermentation phase is approximately 30 percent

of the total, and those in recovery approximately 50 percent. Hence, about 9,000 to 22,500

workers might be expected to hold jobs in the

immediate fermentation area, and about 15,000

to 37,500 workers would be involved in handling the production medium (with or without

the organisms).

Estimates of the number of totally new jobs

that would be created are highly speculative;

they should allow for estimates of increases in

the quantity of chemicals currently being produced and the production of totally new compounds. According to estimates by Genex, the

new and growth markets may reach $26 billion

by the year 2000, which would add 52,000 to

130,000 jobs to the present number.
chapter 6

The Food Processing Industry
Chapter 6

Page

Introduction—The Industry . . . . . . . . . . . . . .. .107

Single-Cell Protein. . . . . . . . . . . . . . . . . . . . . .. .107

Genetic Engineering and SCP Production. ... 109

Commercial Production. . ................109

Genetics in Backing, Brewing, and Winemaking .110

Microbial Polysaccharides. ., . ..............111

Enzymes. . . . . . . . . . . . . . . . . . . . . , . ........111

Genetic Engineering and Enzymes in the

Food Processing Industry . . . . . . . . . . . . . . . 112

Sweeteners, Flavors, and Fragrances . . . . . . . . 112

Overview . . . . . . . . . . . . . . . . . . . . ..........113

Tables

21. Classification of Yeast-Related U.S. Patents , , 108

22. Comparison of Selling Price Ranges for

Selected Microbial, Plant, and Animal

Protein Products. . . . . . . . . . ............108

23, Raw Materials Already Tested on a

Laboratory or Small Plant Scale. ... , , .. ...109

Figure

Figure No. Page

26. The Use of Hybridization To Obtain a Yeast

Strain for the Production of LowCarbohydrate Beer . . . . . . . . . . . . ........110

Table No. Page

20. Estimated Annual Yeast Production, 1977...108
Chapter 6

The Food Processing Industry

Introduction—the industry

The food processing industry comprises

those manufacturers that transform or process

agricultural products into edible products for

market. It is distinguished from the production,

or farming and breeding portions of the agricultural industry.

Genetics can be used in the food processing

industry in two ways: to design micro-organisms that transform inedible biomass into food

for human consumption or for animal feed; and

to design organisms that aid in food processing,

either by acting directly on the food itseIf or by

providing materials that can be added to food.

Eight million to ten million people work in the

meat, poultry, dairy, and baking industries; in

canned, cured, and frozen food plants; and in

moving food from the farm to the dinner table.

In 1979, the payroll was over $3.2 billion for the

meat and poultry industries, $2.6 billion for

baking, and $1.9 billion for food processing.

Single-cell protein

The interest in augmenting the world’s supply of protein has focused attention on microbial sources of protein as food for both animals

and humans. * Since a large portion of each

bacterial or yeast cell consists of proteins (up to

72 percent for some protein-rich cells), large

numbers have been grown to supply single-cell

protein (SCP) for consumption, The protein can

be consumed directly as part of the cell itself or

can be extracted and processed into fibers or

meat-like items. By now, advanced food processing technologies can combine this protein

with meat flavoring and other substances to

produce nutritious food that looks, feels, and

tastes like meat.

*AS an example of the potential significance of SCP, the Soviet

IInion, which is one of the largest producers, expects to produce

enough fodder yeast from internally available raw materials to be

self-sufficient in animal protein foodstuffs by 1990.

Traditionally, micro-organisms have been

used to stabilize, flavor, and modify various

properties of food. More recently, efforts have

been made to control microbial spoilage and to

ensure that foods are free from micro-organisms that may be hazardous to public health.

These are the two major ways in which microbiology has been useful.

Historically, most efforts have been devoted

to improving the ability to control the harmful

effects of micro-organisms. The industry recognized the extreme heat resistance of bacterial

spores in the early 20th century and sponsored

or conducted much of the early research on the

mechanisms of bacterial spore heat resistance.

Efforts to exploit the beneficial characteristics

of micro-organisms, on the other hand, have

been largely through trial-and-error. Strains

that improve the quality or character of food

generally have been found, rather than designed.

The idea of using SCP as animal feed or

human food is not new; yeast has been used as

food protein since the beginning of the century.

However, in the past 15 years, there has been a

dramatic increase in research on SCP and in the

construction of large-scale plants for its production, especially for the production of yeast.

(See table 20.) Interest in this material is reflected in the numerous national and international conferences on SCP, the increasing

number of proceedings and reviews published,

and the number of patents issued in recent

years. (See table 21.)

The issues addressed have covered topics

such as the economic and technological factors

influencing SCP processes, nutrition and safety,

and SCP applications to human or animal foods.

Thus far, commercial use has been limited by

107
108 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Table 20.-Estimated Annual Yeast Production, 1977

(dry tonnes)

Baker’s yeast Dried yeasta

Europe. . . . . . . . . . . . . . . . . . 74,000 b

160,000b

North America . . . . . . . . . . . 73,000 53,000

The Orient. . . . . . . . . . . . . . . 15,000 25,000

United Kingdom. . . . . . . . . . 15,500 (c)

South America. . . . . . . . . . . 7,500 2,000

Africa. . . . . . . . . . . . . . . . . . . 2,700 2,500

Totals . . . . . . . . . . . . . . . . 187,700 242,500

aDrigd yeast includes food and fodder yeasts; data for petroleum-grown Yeasts

are not available.

bproduction figures for U.S.S.R. not reported.

cNone reported.

SOURCE: H. J. Peppier and D. Perlman (eds.), Microbial Technology, vol. 1 (London: Academic Press, 1979), p. 159.

Table 21.—Classification of Yeast-Related

U.S. patents (1970 to July 1977)

Category Number issued

Yeast technology (apparatus, processing) . . . . 22

Growth on hydrocarbons. ., . . . . . . . . . . . . . . . . 28

Growth on alcohols, acids, wastes. . . . . . . . . . . 22

Production of chemicals. . . . . . . . . . . . . . . . . . . 14

Use of baking and pasta products . . . . . . . . . . . 24

Condiments and flavor enhancers . . . . . . . . . . . 18

Reduced RNA. . . . . . . . . . . . . . . . . . . . . , . . . . . . 11

Yeast modification of food products . . . . . . . . . 13

Isolated protein . . . . . . . . . . . . . . . . . . . . . . . . . . 5

Texturized yeast protein . . . . . . . . . . . . . . . . . . . 7

Lysates and ruptured cells . . . . . . . . . . . . . . . . . 7

Animal feed supplements . . . . . . . . . . . . . . . . . . 12

Total. ., . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183

SOURCE: H. J. Peppier, “Yeast,” Annual Report on Fermentation Processes, D.

Perlman (cd.), vol. 2 (London: Academic Press, 1978), pp. 191-200.

several factors. For each bacterial, yeast, or

algal strain used, technological problems (from

the choice of micro-organisms to the use of corresponding raw material) and logistical problems of construction and location of plants have

arisen. But the primary limitation so far has

been the cost of production compared with the

costs of competing sources of protein. (The comparative price ranges in 1979 for selected

microbial, plant, and animal protein products

are shown in table 22.)

The costs of manufacturing SCP for animal

feed in the United States are high, particularly

relative to its major competing protein source,

soybeans, which can be produced with little fertilizer and minimal processing. The easy availability of this legume severely limits microbial

SCP production for animal feed or human food.

In fact, according to the U.S. Department of

Table 22.—Comparison of Selling Price Ranges

for Selected Microbial, Plant, and Animal

Protein Products

Crude Price range

protein 1979 Us.

Product, substrate, and quality content dollars/kg

Single cell proteins

Candida utilis, ethanol, food grade 52 1.32-1.35

Kluyveromyces tragilis, cheese

whey, food grade. . . . . . . . . . . . . 54 1.32

Saccharomyces cerevisiae:

Brewer’s, debittered, food grade 52 1.00-1.20

Feed grade. . . . . . . . . . . . . . . . . . 52 0.39-0.50

Plant proteins

Alfalfa (dehydrated) . . . . . . . . . . . . 0.12-0.13

Soybean meal, defatted. . . . . . . . . 0.20-0.22

Soy protein concentrate. . . . . . . . . 70-72 0.90-1.14

Soy protein isolate . . . . . . . . . . . . . 90“ 92 1.96-2.20

Animal proteins

Fishmeal (Peruvian) . . . . . . . . . . . . 65 0.41-0.45

Meat and bonemeal . . . . . . . . . . . . 50 0.24-0.25

Dry skim milk. . . . . . . . . . . . . . . . . . 37 0.88-1.00

SOURCE: Office of Technology Assessment.

Agriculture (USDA), total domestic and export

supply for U.S. soybeans will grow 73 percent

by 1985.

Soybeans are primarily consumed as animal

feed. But while only 4 percent of their annual

production are directly consumed by humans,

the market is growing significantly. The introduction of improved textured soy protein in

cereals, in meat substitutes and extenders, and

in dairy substitutes has increased the use of soy

products. Nevertheless, the market does not demand soy products in particular but protein

supplements, vegetable oils, feed grain supplements, and meat extenders in general. Other

protein and oil sources could replace soybeans

if the economics were attractive enough. Fishmeal, dry beans, SCP, and cereals are all potential competitors. As long as a substitute can

meet the nutritional, flavor, toxicity, and regulatory standards, competition will be primarily

based on price.

The competition between soybeans and SCP

illustrates one of the paradoxes of genetic engineering. While significant research is attempting the genetic improvement of soybeans, genetic techniques are also being explored to increase the production of SCP. Consequently, the

same tool—genetic engineering—encourages

competition between the two commodities.
Ch. 6—The Food Processing Industry ● 109

Genetic engineering and

SCP production

Despite the microbial screening studies that

have been conducted and the wealth of basic

genetic knowledge available about common

yeast (a major source of SCP), genetic engineering has had little economic impact on SCP processes until recently. Today, a variety of substances are being considered as raw materials

for conversion.

● Petroleum-based hydrocarbons.—Until recently, the wide availability and low cost of

petrochemicals have made the n-alkane hydrocarbons (straight chain molecules of

carbon and hydrogen), which are petrochemical byproducts, potential raw materials for SCP production. At British Petroleum, mutants of micro-organisms have

been obtained having an increased protein

content. Mutants have also been found

with other increased nutritive values, e.g.,

vitamin content.

● Methane or methanol-Relatively few genetic studies have been directed at investigating the genetic control of the microbial

use of methane or methanol. However, one

recent application of genetic engineering

has been reported by the Imperia] Chemical Industries (ICI) in the United Kingdom,

where the genetic makeup of a bacterium

(Methylophilus methylotrophus) has been

altered so that the organism can grow

more readily on methanol. The increase in

growth provides increased protein and has

made its production less expensive. The

genetic alteration was accomplished by

transferring a gene from Escherichia coli to

M. methylotrophus.

● Carbohydrates. —Many carbohydrate substrates—from starch and cellulose to beets

and papermill wastes—have been investigated. Forests are the most abundant

source of carbohydrate in the form of cellulose. But before it can be used by microorganisms, it must be transformed into the

carbohydrate, glucose, by chemical or enzymatic pretreatment. Many of the SCP

processes that use cellulose employ organisms that produce the enzyme cellulase,

which degrades cellulose to glucose.

Most of the significant genetic studies on the

production of cellulase by micro-organisms are

just beginning to appear in the literature. The

most recent experiments have been successful

in creating fungal mutants that produce excess

amounts.

Commercial production

Of the estimated 2 million tons of SCP produced annually throughout the world, most

comes from cane and beet molasses, with about

500,000 tons from hydrolyzed wood wastes,

corn trash, and papermill wastes. (See table 23.)

Integrated systems can be designed to couple

the production of a product or food with SCP

production from wastes. E.g., the waste sawdust from the lumber industry could become a

source of cellulose for micro-organisms. ICI’s

successful genetic engineering of a micro-organism to increase the usefulness of one raw

material (methanol) should encourage similar

attempts for other raw materials.

But while SCP can be obtained from a wide

variety of micro-organisms and raw materials,

the nutritional value and the safety of each

micro-organism vary widely, as do the costs of

competing protein sources in regional markets.

Consequently, accurate predictions cannot be

made about the likelihood that SCP will displace

traditional protein products, overall. Displacements have and will continue to occur on a caseby-case basis.

Table 23.-Raw Materials Already Tested on a

Laboratory or Small Plant Scale

Agave juices

Barley straw

Cassava

Citrus wastes

Date carbohydrates

Meatpacking wastes

Mesquite wood

Peat (treated)

Pulpmill wastes

Sawdust

Sunflower seed husks

(treated)

Wastes from chemical

production of maleic

anhydride

Waste polyethylene (treated)

SOURCE: Office of Technology Assessment.
110 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Genetics in baking, brewing, and winemaking

The micro-organism of greatest significance

in the baking, brewing, and winemaking industries is common yeast. Because of its importance, yeast was one of the first micro-organisms to be used in genetic research. Nevertheless, the surge in studies in yeast genetics has

not been accompanied by an increase in its

practical application, for three reasons:

●

●

●

industries already have the desired efficient strains, mainly as a result of trial-anderror studies;

new genetic strains are not easily bred;

they are incompatible for mating and the

genetic characteristics are poorly understood; and

many of the important characteristics of

industrial microbes are complex; several

genes being responsible for each.

Changing technologies in the brewing industry and increased sophistication in the molecular genetics of yeast have made it possible for

researchers to achieve novel goals in yeast

breeding. one strain that has already been constructed can produce a low-carbohydraate beer

suitable for diabetics. (See figure 26. )

The baking industry is also undergoing technological revolution, and yeasts with new properties are now needed for the faster fermentation of dough. New strains with improved biological activity, storage stability, and yield

would allow improvements in the baking process.

In the past, most genetic applications have

come in the formation of hybrid yeasts. The

newel* genetic approaches, which use cell fusion now open up the possibility of hybrids developed from strains of yeast that carry useful

genes but cannot mate normally,

Classical genetic research has also been carFigure 26.—The Use of Hybridization To Obtain

a Yeast Strain for the Production of

Low-Carbohydrate Beer

SOURCE: Office of Technology Assessment.

ried out with wine yeasts. Interestingly, within

the past 10 years, scientists have isolated induced mutants of wine yeasts that have: I) an

increased alcohol tolerance and the capacity to

completely ferment grape extracts of unusually

high sugar content; 2) improved sedimentation

properties, improving or facilitating separation

of yeasts from the wine; and 3) improved performance in the production of certain types of

wines. Hybridization studies of wine yeasts

have been actively pursued only recently.

Progress in developing strains of yeast with

novel properties is limited by the lack of enough

suitable approved systems for using recombinant DNA (rDNA) technology. Eventual approval by the Recombinant DNA Advisory Committee is expected to boost applied research for the

brewing, baking, and winemaking industries.
Ch. 6—The Food Processing Industry ● 177

Microbial polysaccharides

The food processing industry uses polysaccharides (polymeric sugars) to alter or control

the physical properties of foods. Many are incorporated into foods as thickeners, gelling

agents, and agents to control ice crystal formation in frozen foods. They are used in instant

foods, salad dressings, sauces, whips, toppings,

processed cheeses, and dairy products. New

uses are constantly appearing. The annual market in the United States is reported to be over

36,000 tons, not including starches and derivatives of cellulose.

Since many of the polymeric sugars now used

in food processing are derived from plant

sources, microbial polysaccharides have had

limited use. To compete economically, a microbial polysaccharide must offer new properties,

meet all safety requirements, and be readily

available. Very few have reached the level of

commercial applications; the only one in largescale commercial production is xanthan gum. *

● The history of the development of xanthan gum indicates that

the commercially significant organisms resulted from an extensive

screening program for gum producers stored in the Northern UtiEnzymes

Enzymes are produced for industrial, medical, and laboratory use both by fermentation

processes that employ bacteria, molds, and

yeasts and by extraction from natural tissues.

The present world market for industrial enzymes is estimated to be $l50 million to $174

million; the technical (laboratory) market adds

another $20 million to $40 million. Fewer than

so microbial enzymes are of industrial importance today, but patents have been granted for

more than a thousand. This reflects the increasing interest in developing new enzyme prodA wide variety of polysaccharides could theoretically be produced for use in food processing,

Applied genetics may increase their production,

modify those that are produced, eliminate the

degradative enzymes that break them down, or

change the microbes that produce them. However, as with other microbial processes, the application of genetics depends on an understanding of both the biochemical pathway for synthesis of a given polysaccharide and the systems

that control microbial production. For many microbial polysaccharides, this information does

not yet exist; furthermore, little is known about

the enzymes that may be used to modify poly -

saccharides to more useful forms. Progress will

only be able to occur when these information

gaps are filled.

Iization Research and Oeveiopment Dii’ision ol LISL)A’S large nlicrohia! culture co]]ection. Xanthiin gunl produced h+v Xanlhomonas campestris NRRI, B-1459 was found to ha\re chiiracterist ics t hilt

rendered it very promising as a commercial product. in 1960, the

Kelco division of Merck &. (:0., inc., carried out pilot Plii[lt feasihili -

t,v studies, and substantial commercial production t)e~iin in 1964.

Although much Of the work to diit~ hiis I}(+pll (’ii].ri[d OLII wiih

polvsaccharides from one Pitrti[;uliir stritin, t here is increilsing e\idence to suggest thilt ihey could iilso he produced from other

S1 rains.

ucts; it also shows that it is easier to discover a

new enzyme than to create a profitable application for it. *

Most industrial enzymes are used in the detergent industry and the food processing in-

● The enzyme literature is extensive i~tld comprises well over

10,00( papers per year. Although less thitll 5(1 percent of these

publications are concerned with microhial enzymes iind mos[ are

found to have no industrial interest, iI few Ihousand papers per

year are of potential interest for the industriill development of enzymes. Less than 100 papers dealing with industrial processes appear every year, and few desct~be processes of great economic significance.
112 ● Impacts of Applied Genetics —Micro-Organisms, Plants, and Animals

dustry, particularly for starch processing. Enzymes began to be used in quantity only 20

years ago. In the early 1960’s, glucoamylase enzyme treatment began to replace traditional

acid treatment in processing starch; around

1965, a stable protease (an enzyme) was introduced into detergent preparations to help

break down certain stains; and in the 1970’s,

glucose isomerase was used to convert glucose

to fructose, practically creating the high-fructose corn syrup industry.

Genetic engineering and enzymes in

the food processing industry

Biotechnology applied to fermentation processes will make available larger quantities of existing enzymes as well as new ones. (See ch. 5.)

The role of genetic engineering in opening commercial possibilities in the food processing industry is illustrated by the enzyme, pullulanase.

This enzyme degrades pullulan, a polysaccharide, to the maltose or high-maltose syrups that

give jams and jellies improved color and brilliance. They reduce off-color development produced by heat in candies and prevent sandiness

in ice cream by inhibiting sugar crystallization.

Mahose has several unique and favorable characteristics. It is the least water-absorbent of the

maltose sugars and, although it is not as sweet

as glucose, it has a more acceptable taste. It is

also fermentable, nonviscous, and easily soluble. It does not readily crystallize and gives desirable browning reactions.

Pullulanase can also break down another carbohydrate, amylopectin, to produce high amy -

lose starches. These starches are used in industry as quick-setting, structurally stable gels, as

binders for strong transparent films, and as

coatings. Their acetate derivatives are added to

textile finishes, sizing, adhesives, and binders.

In food, amylose starches thicken and give texture to gumdrop candies and sauces, reduce fat

and grease in fried foods, and stabilize the protein, nutrients, colors, and flavors in reconstituted products like meat analogs.

In view of the current shortages of petroleum-derived plastics and the need for a biodegradable replacement, amylose’s ability to form

plastic-like wraps may provide its largest industrial market, although that market has not yet

been developed.

[f applications for the products made by

pullulanase can be developed, genetic engineering can be used to insert this enzyme into industrially useful organisms and to increase its

production. However, the food processing industry is permitted to use only enzymes that are

obtained from sources approved for food use.

Since the chief source of pullulanase is a pathogenic bacterium, KlebsiellaII aerogenes, no significant efforts have been made to apply genetics to

improve its production or quauality. Molecular

genetics could ultimately transfer the pullulanase trait from K. aerogenes to a micro-organism

approved for food use, if approved micro-organisms that manufacture pullulanase cannot be

found.

Sweeteners, flavors, and fragrances

Biotechnology has already had a marked impact on the sweetener industry. The availability

of the enzymes glucose isomerase, invertase,

and amylase has made the production of highfructose corn sweeteners (HFCS) profitable. Production of HFCS in the United States has increased from virtually nothing in 1970 to 10

percent of the entire production of caloric

sweeteners in 1980 (11 lb per capita). The price

advantage of HFCS is expected to cause its continued growth, particularly in the beverage industry. In fact, the Coca Cola Co. announced in

1980 that fructose will soon constitute as much

as 50 percent of the sweetener used in its name

brand beverage.

Biotechnology can be used to produce other

sweeteners as well. While it is unlikely that sucrose will ever be made by micro-organisms (although improvements in sugarcane and sugar

beet yields may result from agricultural genetic

studies, see ch. 8.), the microbial production of

low-caloric sweeteners is a distinct possibility.

Three new experimental sweeteners—aspartame, monellin, and thaumatin—are candidates.

Aspartame is synthesized chemically from

the amino acids, aspartic acid and phenylalanine, which can themselves be made by fermen-
Ch. 6— The Food Processing Industry ● 113

tation. The possibility of using microbes to COUple the two amino acids is being investigated in

at least one biotechnology research firm. Chemical production of aspartame is expensive and

benefits from biotechnology are possible.

Monellin and thaumatin are natural substances—proteins obtained from West African

plants. Both are intensely sweet–up to 100,000

times sweeter than table sugar—and the sensation of sweetness can last for hours. Their

microbial production may be competitive with

their extraction from plants. Since the physical

and biological properties of thaumatin are

known, it might also be produced through genetic engineering. Such an approach would not

only increase the available supply, but would

offer new molecules for investigating the physiology of taste.

Other flavors and fragrances show less promise at present. Although the chemistry of several flavors and aromas has been identified, too

little research into their use has been conducted. *

● KW’elll ttr(}l’k 011 1116? 101’lllilliOll 1)}1 Illi(’1’()-{)l’gilllisllls ot” tliii’01’

illld ill’olllil (’h~llli(’ill S hllol$’11 iiS lil(’lollt>S iill(l terpfmoids tlil S I)(WI1

I’(?p{)l’td. [.il[Ytoll~S 0(:(:111” ilS !]iil’ol’-(’oll~ l’lt)lltlll~” (’Olllpollf?lltS il l

IIUII1.V l’~1’il)t?tltiitiotl produ(’ts, \\’h[’l’(> III(’}I ii[’(~ lk)IvN(d I)v microhial

reiictions. [)it’t’erent ])i~th~iii.vs exist !(N- t heir n~icrohial i“ornlation.

k;.g., galllllla-t)llty l’ola{’toll[” , mfhich is I“ol’llled during J~CilSt ternlentittion, is 1(}uIM1 in sherry, wine, iind hf?er. /IS eiIt.1.v iis Ig3(J, an ot’-

giinism Wiis isolated f]xNll orange leiii,t?s that hii{j a pea(+-lik(+ ~d(]r

iimi Wiis thougll[ to he Sporobdonl.vces resew. The kwtones, ZIdecmolide ii[ld (:is-6-docle(’ell-4-olici(’” Mere found to he responsiI)le.

Overview

The application of genetic engineering will affect the food processing industry in piecemeal

fashion. Isolated successes can be expected for

certain food additives, such as aspartame (not

yet approved by the Food and Drug Administration (FDA) for sale in the United States) and fructose, and for improvements in SCP production.

But an industrywide impact is not expected in

the near future because of several conflicting

forces:

●

●

●

The basic genetic knowledge of characteristics that could improve food has not been

adequately developed.

The food processing industry is conservative in its research and development expenditures for improved processes, generally allocating less than half as much as

more technologically sophisticated industries.

Products made by new microbial sources

must satisfy FDA safety regulations, which

include undergoing tests to prove lack of

harmful effects. * It may be possible to re-

“ fi;.g., all food additites and nli(’1.()-{)1.gar)isr~ls used in food procmsing must he approtwd its geIWriill&V regarded iIs sate.

duce the amount of required testing by

transferring the desired gene into microorganisms that already meet FDA standards.

Nevertheless, the application of new genetic

technologies will probably accelerate. Technologically sophisticated companies are being

drawn into the business. Traditionally capitalintensive companies such as Union Carbide,

ITT, General Electric, Corning Glass, and

McDonnell-Douglas can be expected to introduce automation and more sophisticated engineering to food processing, modernizing the industry’s technology. As

industry observer: 1

You don’t work on a

fish. You try to change

has been noted by one

better way to preserve

the system so that you

no longer catch fish; you “manufacture” them

and, if possible, do it right on top of your market so that you don’t have to preserve them at

all.

IM. L. Kastens,

1980, pp. 215-217.

“The Coming Food Industry,” Chemtech, April
114 ● Impacts of Applied Genetics-Micro-Organisms, Plants, and Animals

You don’t worry about processing bacon Genetic engineering can be expected to aid in

without nitrites, you engineer a synthetic bacon the creation of novel food preparations through

with designed-in shelf life. effects on both the food itself and the additives

You don’t try to educate people to eat a “bal- used for texturizing, flavoring, and preserving.

anced diet;” you create a “whole” food with the

proper balance of nutrients and supplements,

and you make it taste like something people

already like to eat.
Chapter 7

The Use of Genetically

Engineered Micro-Organisms

in the Environment
Chapter 7

Page

Mineral Leaching and Recovery . . . . . . . . . . , . . 117

Microbial Leaching . . . . . . . . . . . . . . . . . . . . . 117

Applied Genetics in Strain Improvement . . , . . 118

Metal Recovery . . . . , . . . . . . . . . . . . . . . . . . . 118

Oil Recovery . . . . . . . . . . . . . . . . . . . . . . . . . .. .119

Enhanced Oil Recovery . . . . . . . . . . . . . . . . . . 119

Microbial Production of Chemicals Used

in EOR . . . . , . . . . . . . . . . . . . , . . . . . , . , , 120

In Situ Use of Micro-Organisms. . . , . . . . . . . 121

EOR and Genetic Engineering . ...........122

Constraints to Applying Genetic

Engineering Technologies in EOR. . ......122

Genetic Engineering of Micro-Organisms

for Use in other Aspects of Oil Recovery

and Treatment. . . . . . . . . . . . . . . . . . . . . . 123

Overview of Genetic Engineering in Mining

and Oil Recovery . . . . . . . . . . . . . , . . . . . . 123

Page

Pollution Control. . . . . . . . . . . . . . . . . . . . . . . . . 123

Enhancing Existing Microbial Degradation

Activity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.24

Adding Microbes to Clean Up Pollution . . . . . . 124

Commercial Applications—Market Size and

Prospects . , . . . . . , . . . . . . . . . . . . . . . . . . . 125

Genetic Research in Pollution Control . . . . . . . 126

Federal Research Support for Engineering

Microbes to Detoxify Hazardous Substances . 127

Summary. . . . . . . , . . . . , . . . . . . . . . . . . . . . . 127

Issue and Options—Biotechnology, . . . . . . . . . . . 128

Figure

Figure No. Page

27. chemical Flooding Process . . . . . . . . . . . . . . 120
Chapter 7

The Use of Genetically Engineered

Micro-Organisms in the Environment

Although most genetically engineered micro- ●

organisms are being designed for contained facilities like fermenters, some are being exam- ●

ined for their usefulness in the open environ- ●

ment for such purposes as mineral leaching and

recovery, oil recovery, and pollution control.

All three applications are characterized by:

● the use of large volumes of micro-organisms;

less control over the behavior and fate of

the micro-organisms;

a possibility of ecological disruption; and

less basic research and development (R&D)

—and a higher degree of speculation—than

the industries previously discussed.

Mineral leaching and recovery

All micro-organisms interact with metals,

Two interactions that are of potential economic

and industrial interest are leaching metals from

their ores, and concentrating metals from

wastes or dilute mixtures. The first would allow

the extraction of metals from large quantities of

low-grade ores; the second would provide methods for recycling precious metals and controlling pollution caused by toxic metals.

Microbial leaching

In microbial or bacterial leaching, metals in

ores are made soluble by bacterial action. Even

before bacterial leaching systems became accepted industrial practice, it was known that

dissolved metals could be recovered from mine

and coal wastes. Active mining operations currently based on this process (such as those in

Rio Tinto, Spain) date back to the 18th century.

Presently, large-scale operations in the United

States use bacterial leaching to recover copper

“rem waste material. Estimates for the contribution of copper leaching to the total annual

J.S. production range from 11.5 to 15 percent.

Leaching begins with the circulation of water

through large quantities–often hundreds of

ins—of ore. Bacteria, which are naturally assoiated with the rocks, then cause the metals to

be leached by one of two general mechanisms:

either the bacteria act directly on the ore to extract the metal or they produce substances,

such as ferric iron and sulfuric acid, which then

extract the metal. It appears that simply adding

acid is not as efficient as using live bacteria.

Although acid certainly plays a role in metal extraction, it is possible that direct bacterial attack

on some ores is also involved. In fact, some of

the bacteria that are known to be involved in

mineral leaching have been shown to bind tenaciously to those minerals.

The application of the leaching process to

uranium mining is of particular interest because of the possibility of in situ mining. Instead

of using conventional techniques to haul uranium ore to the surface, microbial suspensions

can extract the metal from its geological setting.

Water is percolated through underground

shafts where the bacteria dissolve the metals.

The solution is then pumped to the surface

where the metal is recovered. This approach,

also called “underground solution mining, ” is

already used in Canadian uranium mines,

where it began almost by chance. In 1960, after

only 2 years of operation, researchers at the

Stanrock Uranium Mine found that the natural

underground water contained large amounts of

leached uranium. In 1962, over 13,000 kilo117

76-565 0 - 81 - 9
118 ● Impacts of Applied Genetics —Micro-Organisms, Plants, and Animals

grams (kg) of uranium oxide were obtained

from the water. Thereafter, water was circulated through the mines as part of the mining

operation. It has been suggested that extending

this practice to most mines would have significant environmental benefits because of the

minimal disruption of the land surface.

Although the process is slower than the technology currently employed, the operating costs

might be lower because of the simplicity of the

system, since no grinding machinery is needed.

Furthermore, deeper and lower grade deposits

could be mined more readily.

Bacterial leaching can also extract sulfur-containing compounds, such as pyrite, from coal,

producing coal with a lower sulfur content.

Sulfur-containing coals from such areas as Ohio

and the Appalachian Mountains are now less desirable than other coals because of the sulfur

dioxide they release during burning. They often

contain up to 6 percent sulfur, of which 70 percent can be in the form of pyrite. According to

recent data, mixed populations of different bacteria, rather than a single species, are responsible for the most effective removal of sulfur—a

finding that may lead to the genetic engineering

of a single sulfur-removing bacterium in the

future.

Applied genetics in strain improvement

The bacterium most studied for its leaching

properties has been Thiobacillus ferrooxidians

(which leaches copper), but others have also

been identified in natural leaching systems.

Although leaching ability is probably under

genetic control in these organisms, practically

nothing is known about the precise mechanisms. This is largely because little information

exists in two critical areas: the chemistry of interaction between the bacteria and rock surfaces; and the genetic structure of the microorganisms. The finding that mixed populations

of bacteria interact to increase leaching efficiency complicates the investigation.

Because of the lack of genetic and biochemical information about these bacteria, the application of genetic technologies to mineral leaching remains speculative. Progress in obtaining

more information is slow because less than a

dozen laboratories in the Nation are actively

performing research.

But even when the scientific knowledge is

gathered, two obstacles to the use of genetically

engineered micro-organisms will remain. The

first is the need to develop engineered systems

on a scale large enough to exploit their biological activities. A constant interchange must take

place between microbial geneticists, geologists,

chemists, and engineers. E.g., the geneticists

must understand the needs identified by the

geologists as well as the problems faced by the

engineers, who must scale-up laboratory-scale

processes. The complex nature of the problem

can be approached most successfully by an

interdisciplinary group that recognizes the

needs and limitations of each discipline.

The second obstacle is environmental. Introducing large numbers of genetically engineered micro-organisms into the environment

raises questions of possible ecological disruption, and liability if damage occurs to the environment or human health.

In summary, the present lack of sufficient

scientific knowledge, scientists, and interdisciplinary teams, and the concerns for ecological

safety present the major obstacles to the use of

genetic engineering in microbial leaching.

Metal recovery

The use of micro-organisms to concentrate

metals from dilute solutions such as individual

waste streams has two goals: to recover metals

as part of a recycling process; and to eliminate

any metal that may be a pollutant. The process

makes use of the ability of micro-organisms to

bind metals to their surfaces and then concentrate them internally.

Studies at the Oak Ridge National Laboratory

in Tennessee have shown that micro-organisms

can be used to remove heavy metals from industrial effluents. Metals such as cobalt, nickel,

silver, gold, uranium, and plutonium in concen

trations of less than 1 part per million (ppm) car.

be recovered. The process is particularly useful

for recovering metals from dilute solutions 01
Ch. 7—The Use of Genetically Engineered Micro-Organisms in the Environment ● 119

10 to 100 ppm, where nonbiological methods As with other biological systems, genetic

may be uneconomical. Organisms such as the engineering may increase the efficiency of the

common yeast Saccharomyces cerevisiae can ac- extraction process. In the Saccharomyces syscumulate uranium up to 20 percent of their tern, differences in the ability to recover the

total weight. metals have been demonstrated within popula- —

tions of cells. Selection for cells with the genetic The economic competitiveness of biological ability to accumulate large amounts of specific, methods has not yet been proven, but genetic desired metals would be an important step in improvements have been attempted only re- designing a practical system. cently. The cost of producing the micro-organisms has been a major consideration. If it can

be reduced, however, the

useful.

Oil recovery

approach might be

Since 1970, oil production in the United

States has declined steadily. The supply can be

increased by: accelerating explorations for new

oilfields; by mining oil shale and coal and converting them to liquids; and by developing new

methods for recovering oil from existing reservoirs.

In primary methods of oil recovery, natural

expulsive forces (such as physical expansion)

drive the oil out of the formation. In secondary

methods of recovery, a fluid such as water or

natural gas is injected into the reservoir to force

the oil to the well. Approximately 50 percent of

domestic crude in recent years has been obtained through secondary recovery.

Recently, new methods of oil recovery have

been added to primary and secondary methods,

which are called tertiary, improved, or enhanced oil recovery (EOR) techniques. They employ chemical and physical methods that increase the mobility of oil, making it easier for

other forces to drive it out of the ground. The

major target for EOR is the oil found in sandstone and limestone formations. It is here that

applied genetics may play a major role,

engineering micro-organisms to aid in recovery.

Oil susceptible to these processes is localized

in reservoirs and pools at depths ranging from

100 ft to more than 17,000 ft. In these areas, the

oil is adsorbed on grains of rock, almost always

accompanied by water and natural gas. The

physical association of the trapped oil and the

surrounding geological formations varies significantly from site to site. The unknown characteristics of these variations are largely responsible for the economic risk in an attempted EOR.

Enhanced oil recovery

Of the original estimated volume of more

than 450 billion barrels (bbl) of U.S. oil reserves,

about 120 billion bbl have been recovered by

primary and secondary techniques, and another

30 billion bbl are still accessible by these

methods. The remaining 300 billion bbl however, are probably recoverable only by EOR

methods. These figures include the oil remaining in known sandstone and limestone reservoirs and exclude tar sands and oil shale.

Four EOR processes are currently used. All

are designed to dislodge the crude oil from its

natural geological setting:

●

●

In thermal processes, the oil reservoir is

heated, which causes the viscosity of the oil

to decrease, and with the aid of the

pressure of the air introduced, supports

the combustion that forces the petroleum

to the producing well. Thermal processes

will not be improved by genetic technologies.

Various crude oils differ in their viscosity—

ability to flow. Primary and secondary

methods can easily remove those that flow
120 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

as readily as water, but many of the reservoirs contain oil as viscous as road tar.

Miscible processes use injected chemicals

that blend with the crude oil to form mixtures that flow more readily. The chemicals used include alcohols, carbon dioxide,

petroleum hydrocarbons such as propane

and butane-propane mixtures, and petroleum gases. A fluid such as water is generally

used to push a “slug” of these chemicals

through the reservoir to mix with the

crude oil and move it to the surface.

● Chemicals are also used in alkaline flooding, polymer flooding, and combined surfactant/polymer flooding.

In alkaline flooding, sodium hydroxide, sodium carbonate, or other alkaline materials are

used to enhance the flow of oil. Neither natural

nor genetically engineered micro-organisms are

considered useful in this process.

Polymer flooding is a recent apparently successful method of recovery. It depends on the

ability of certain chains of long molecules,

known as polymers, to increase the viscosity of

water. Instead of altering the characteristics of

the crude oil, the aim is to make the injected

water more capable of displacing it.

In the combined surfactant/polymer flooding

technique, a detergent-like material (surfactant)

is used to loosen the oil from its surrounding

rock, while water that contains a polymer to increase its viscosity is used to drive the oil from

the reservoir. (See figure 27.)

● Other EOR methods include many novel

possibilities, such as the injection of live

micro-organisms into a reservoir. These

may produce any of the chemicals used in

miscible and chemical processes, from surfactants and polymers to carbon dioxide.

One target for EOR is the half million stripper wells (producing less than 10 barrels

per day (bbl/d) in the United States.

MICROBIAL PRODUCTION OF CHEMICALS

USED IN EOR

EOR methods that use chemicals tend to be

expensive because of the cost of the chemicals.

Nevertheless, potentially useful polymers were

Figure 27.-Chemical Flooding Process

injection fluids Oil and water

4

Legend I I

zone

SOURCE: Office of Technology Assessment, Enhanced Oil Recovery Potential

in the United States (Washington, D. C.: U.S. Government Printing Office, January 1978).

found in the early 1960’s and have since been

responsible for the recovery of more than 2

million bbl. Polymers such as polyacrylamide

and xanthan gum can increase the viscosity of

water in concentrations as low as one part in a

thousand. Xanthan gum is readily made in large

quantities by micro-organisms. Different strains

of Enterobacler aerogenes produce a wide variety of other polymers. A useful biopolymer—one

formed by a biological process—might be designed specifically to improve oil recovery.

Xanthan gum, produced by Xanthomonas

campestris and currently marketed by the Kelco

division of Merck & Co., Inc., is useful but far

from ideal for oil recovery. While it has excellent viscous properties, it is also very expensive. Furthermore, unless it is exceptionally

pure, it can plug reservoir pores, since the fluid

often has to travel through hundreds of meters

of fine pores. To avoid such plugging, the fluid

must be filtered to remove bacterial debris before it is injected.

Nevertheless, micro-organisms can be selected or genetically engineered to overcome

many obvious difficulties. * With improved

properties, polysaccharides (polymeric sugars)

● A good organism, for example, might have the following

desired properties: nonpathogenic to humans, plants, or animals;

rapid growth on simple, cheap raw materials; ease of separation

from its products; limited detrimental effect on reservoirs, e.g.,

plugging; easy disposal of cells, e.g., byproduct credits; ability to
Ch. 7—The Use of Genetically Engineered Micro-Organisms in the Environment . 121

obtained by microbial fermentation could compete with those obtained from alternative

sources, especially seaweed. Controlled fermentation is not affected by marine pollution and

weather, and production could be geared to

market demand.

Biological processes have disadvantages primarily in the costs of appropriate raw materials

and in the need for large quantities of solvent.

Current efforts to find cheaper raw materials,

such as sugar beet pulp and starch, show promise. The need for solvents to precipitate and concentrate the polymers before shipment from

plant to field can be circumvented by producing

them onsite.

Micro-organisms can also produce substances

like butyl and propyl alcohols that can be used

as cosurfactants in EOR. It has been calculated

that if n-butanol were used to produce crude oil

at a level of 5 percent of U.S. consumption, 2

billion to 4 billion lb per year—or four to eight

times the current butanol production—would

be required. Micro-organisms capable of producing such surfactants have been identified,

and genetically superior strains were isolated

several decades ago at the Northern Regional

Research Laboratories in Illinois. Other chemicals, such as alcohols that increase the rate of

formation and stability of chemical/crude oil

mixtures and the agents that help prevent precipitation of the surfactants, have also been produced by microbial systems.

The uncertainties of the technical and economic parameters are compounded by the lack

of sufficient field experiments. Laboratory tests

cannot be equated with conditions in actual oil

wells. Each oil field has its own set of characteristics—salinity, pH (acidity and alkalinity),

temperature, porosity of the rock, and of the

crude oil itself—and an injected chemical behaves differently in each setting. In most cases,

not enough is known about a well’s characteristics to predict the nature of the chemical/crude

oil interaction and to forecast the efficiency of

oil recovery.

use water available at site; growth under conditions that discourage the growth of unwanted micro-organisms; no major problems

in culturing the bacterium; and genetic stability.

IN SITU USE OF MICRO-ORGANISMS

One alternative to growing micro-organisms

in large fermenters then extracting their chemical products and injecting them into wells, is to

inject the micro-organisms directly into the

wells. They could then produce their chemicals

in situ.

Unfortunately, the geophysical and geochemical conditions in a reservoir seldom favor the

growth of micro-organisms. High temperature,

the presence of sulfur and salt, low oxygen and

water, extremes of pH, and significant engineering hurdles make it difficult to overcome

these limitations. The micro-organisms must be

fed and the microenvironment must be carefully adjusted to their needs at distances of hundreds to thousands of feet. The oil industry has

already had discouraging experiences with

micro-organisms in the past. In the late 1940’s,

for instance, the injection of sulfite-reducing

micro-organisms, along with an inadvertently

high-iron molasses as a carbon source, resulted

in the formation of iron sulfide, which clogged

the rock pores. One oil company developed a

yeast to break down petroleum, but the size of

the yeast cells (5 to 10 micrometers, um) was

enough to clog the 1-um pores.

Nevertheless, information from geomicrobiology suggests that this approach is worth pursuing. Preliminary field tests have also been encouraging. The injection of 1 to 10 gal of Bacillus

or Clostridium species, along with a watersuspended mixture of fermentable raw materials such as cattle feed molasses and mineral

nutrients, has resulted in copious amounts of

carbon dioxide, methane, and some nitrogen in

reservoirs. The carbon dioxide made the crude

less viscous, and the other gases helped to

repressurize the reservoir. In addition, large

amounts of organic acids formed additional carbon dioxide through reactions with carbonate

minerals. The production of microbial surfactants further aided the process.

Although previous assessments have argued

that reservoir pressure is a significant hindrance to the growth of micro-organisms, more

recent studies indicate the contrary. The microorganisms must, however, be selected for increased salt and pH tolerance.
122 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

EOR AND GENETIC ENGINEERING

The current research approach, funded by

the Department of Energy (DOE) and, independently, by various oil companies, is a two-phase

process. The first phase is to find a microorganism that can function in an oil reservoir

environment with as many of the necessary

characteristics as possible. The second is to alter

it genetically to enhance its overall capability.

The genetic alteration of micro-organisms to

produce chemicals used in EOR has been more

successful than the alteration of those that may

be used in situ. * However, recombinant DNA

(rDNA) technology has not been applied in either category. All efforts have employed artificially induced or naturally occurring mutations.

CONSTRAINTS TO APPLYING GENETIC

ENGINEERING TECHNOLOGIES IN EOR

The genetic data base for micro-organisms

that produce useful polysaccharides is weak.

Few genetic studies have been done. Hence, theoretically plausible approaches such as transferring enzyme-coding plasmids (see ch. 2) for

polysaccharide synthesis, cannot be seriously

contemplated at present. Only the crudest

methods of genetic selection for desirable properties have been used thus far. They remain the

only avenue for improvement until more is

learned about the micro-organism’s genetic

mechanisms.

The biochemical data base for the characteristics of both the micro-organisms and their

products is also lacking. The wide potential for

chemical reactions carried out by microbes remains to be explored. At the same time, a system must be devised to allow easy characterization, classification, and comparison of products

derived from a variety of micro-organisms.

The physical data base for oil reservoirs is

limited. The uniqueness of each reservoir suggests that no universal micro-organism or method of oil recovery will be found. Compounding

● Some of the goals have been to: improve polymer properties to

enhance their commercial applicability; improve polymer production (a major mistake has been to reject a micro-organism in the

initial screening because its level of production was too low); improve culture characteristics, e.g., resistance to phage, rapid

growth, ability to use cheaper raw materials; and eliminate enzymes that naturally degrade the polymers.

this problem is the lack of sufficient physical,

chemical, and biological information about the

reservoirs, without which it is difficult to see

how a rational genetic scheme can be constructed for strains. Clearly, the activities of

micro-organisms under specified field conditions cannot be studied unless researchers

know what the appropriate conditions are.

Three institutional obstacles exist. First, publication in this field is limited because most research is carried out in the commercial world

and remains largely confidential. Second, neither the private nor the public sector has been

enthusiastic about the potential role of microorganisms in EOR, The biological approach has

only recently been given consideration as a way

to advance the state of the art of the technology,

and most oil companies still have limited staffs

in microbiology. To date, DOE’s Division of

Fossil Fuel Extraction has conducted the main

Federal effort. Third, any effort to use microorganisms must be multidisciplinary in nature.

Geologists, microbiologists (including microbial

physiologists and geneticists), chemists, and

engineers must interact to evolve successful

schemes of oil recovery. Thus far, such teams

do not exist.

Environmental and legal concerns have also inhibited progress. Microbial EOR methods usually require significant quantities of fresh water

and thus may compete with municipal and agricultural uses. Furthermore, the use of microorganisms introduces concerns for safety. All

strains of Xanthomonas, which produce xanthan

gum polymer, are plant pathogens. Other

micro-organisms with potential, such as Sclerotium rolfii and various species of Aureobasidium

have been associated with lung disease and

wound infections, respectively.

Immediate environmental and legal concerns,

therefore, arise from the potential risks associated with the release of micro-organisms into

the environment. When they naturally cause

disease or environmental disruption, their use is

clearly limited. And when they do not, genetic

engineering raises the possibility that they

might. Such concerns have reduced the private

sector’s enthusiasm for attempting genetic
Ch. 7—The Use of Genetically Engineered Micro-Organisms in the Environment . 123

engineering. (See ch. 10 for a more detailed

discussion of risk.)

GENETIC ENGINEERING OF MICRO-ORGANISMS

FOR USE IN OTHER ASPECTS OF OIL

RECOVERY AND TREATMENT

Two other aspects of microbial physiology

deserve attention: the microbial production of

oil muds or drill lubricants, and the treatment

of oil once it has been recovered. Drilling muds

are suspensions of clays and other materials

that serve both to lubricate the drill and to

counterbalance the upward pressure of oil. Microbially produced polysaccharides have been

developed for this use. Exxon holds a patent on

a formulation based on the production of xanthan gum, from Xanthomonas campestris, while

the Pillsbury Co. has developed a polysaccharide (glucan) from various species of Sclerotium. At least two of the small genetic

engineering firms have begun research programs to develop biologically produced polysaccharides with the desired lubricant qualities.

Interest in the postrecovery microbial treatment of oil after its extraction centers around

the ability of micro-organisms to remove undesirable constituents from the crude oil itself.

As an indication of recent progress, three distinct microbial systems have been developed to

help remove aromatic sulfur-containing material, a major impurity.

Overview of genetic engineering in

mining and oil recovery

The underlying technical problem with the

use of genetically engineered organisms in

either mining or oil recovery is the magnitude

of the effort. In both cases, large areas of land

and large volumes of materials (chemicals, fluids, micro-organisms) must be used. The results

of testing any new micro-organism in a laboratory cannot automatically be extrapolated to

large-scale applications. The change in

magnitude is further complicated by the lack of

rigid controls. Unlike a large fermenter whose

temperature, pH, and other characteristics can

be carefully regulated, the natural environment

cannot be controlled. Nevertheless, despite the

formidable obstacles, the potential value of the

products in these areas assures continuing efforts.

Pollution control

Life is a cycle of synthesis and degradation—

synthesis of complex molecules from atoms and

simple molecules and degradation by bacteria

yeast, and fungi, back to simpler molecules and

atoms when organisms die. The degradation of

complex molecules is an essential part of life.

Without it, “. . . we’d be knee-deep in dinosaurs. ”1 A more quantitative statement is equally thought provoking. Livestock in the united

States produce 1.7 billion tons of manure annually. Almost all of it is degraded by soil microorganisms.

For a long time people have exploited microbial life forms to degrade and detoxify human

sewage. Now, on a smaller scale, science is

‘R. B. Grubbs, “Bacterial Supplementation, What It Can and Cannot Do. ” oral presentation to the Ninth Engineering Foundation on

Environmental Engineering in the Food Processing Industry, 1979

(Available from Flow Laboratories, Inc., Rockville, Md.)

beginning to use micro-organisms to deal with

the pollution problems presented by industrial

toxic wastes. Chemicals in their place can be

useful and beneficial; out of place, they can be

polluting.

Pollution problems can be divided into two

categories: those that have been present for a

long time in the biosphere-e. g., most hydrocarbons encountered in the petroleum industry

and human and animal wastes—and those that

owe their origin to human inventiveness-e. g.,

certain pesticides. Chemicals of both sorts,

through mishap, poor planning, or lack of

knowledge at the time of their application

sometimes appear in places where they are

potentially or actually hazardous to human

health or the environment.

Pollution can be controlled by microbes in

two ways: by enhancing the growth and activity
124 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

of microbes already present at or near the site

of the pollution problem, and by adding more

(sometimes new) microbes to the pollution site.

The first approach does not provide an opportunity for applying genetics, but an example will

indicate how it functions.

Enhancing existing microbial

degradation activity

Sun Oil successfully exploited indigenous

microbes to clean up a 6,000 gal underground

gasoline spill that threatened the water supply

of a town in Pennsylvania. z 3 First, engineers

drilled wells to the top of the water table and

used pumps to skim gasoline from the water

surface. About half the gasoline was removed in

this fashion, but company calculations showed

that dissipating the remaining gasoline would

require about 100 years. To speedup the process, it was decided to encourage the growth of

indigenous bacteria that could degrade the

gasoline.

Pollution-control microbes, like all organisms,

require a number of different elements and

compounds for growth. If the amount of any

nutrient is limited, the microbe will not be able

to metabolize the pollutant at the fastest rate.

The cleanup depended on increasing the

growth rate of the bacteria by supplying them

with additional nutrients. In the case of the

gasoline-degrading bacteria, the gasoline already supplied the hydrocarbon, but the watergasoline environment was deficient in nitrogen,

phosphate, and oxygen. Those three nutrients

were pumped down to the water table, bacterial

growth increased, and the gasoline was metabolized into innocuous chemicals by the bacteria.

As a result, it was degraded in a single year.

Adding microbes to clean up pollution

Genetics may have important applications in

approaches to pollution control that depend on

‘R. L. Raymond, V. W. Jamison, J. O. Hudson, “Beneficial Stimulation of Bacterial Activity in Groundwaters Containing Petroleum

Products,” AIChE symposium series 73:390404, 1976.

W. W. Jamison, R. L. Raymond, J. O. Hudson, “Biodegradation of

High-Octane Gasoline, ” Proceedings of the Third International Biodegradation Symposium, J. M. Sharpley and A. M. Kaplan (eds.)

(City????: Applied Science Publishers, 1976).

adding microbes to the pollution site. Three

firms–Flow Laboratories, Polybac Corp., and

Sybron/Biochemicals Corp.—sell microbes for

such use. Two companies select bacteria for enhanced degradation activity and two mutate

bacteria to the same end, but none of the three

firms currently uses genetic engineering techniques.

Some “formulations” (mixtures) of bacteria

are designed to degrade particular pollutants,

such as one that was used to digest the 800,000

gal of oily water that lay in the bilges of the

Queen Mary. After a 6-week treatment with the

formulation, the water from the bilges was

judged safe for disposal into the Long Beach,

Calif., harbor. It was discharged without causing an oil slick or harming marine life. q Flow

Laboratories markets its services to companies

with industrial pollution problems. It investigates the problem, develops a formulation to

degrade the pollutants, and sells it.

In addition to industrial pollution problems,

Flow markets its products and services for use

in sewerage systems, which collect and hold

human wastes to facilitate degradation and detoxification. Sludge bacteria in sewerage plants

degrade the waste, but they are not present in

the lines that carry wastes to the treatment

plant. As a result, greases and oils from fat discarded through garbage disposals and from cosmetic oils and creams coat the inside of sewerage lines and reduce their carrying capacity.5

Cities have resisted using added microbes in

sewerage systems. Standard textbooks simply

state that the ideal bacteria will establish themselves in a well-planned and well-managed system. The idea that “better” bacteria can be

added to improve the plant operation is not

readily accepted.

The value of adding bacteria to large sewerage sytems has not been adequately tested.

Because of the size of municipal systems (which

already contain tons of sludge bacteria), some

have argued that adding a few additional

4Anon., Environmental Science and Technology 13:1180, 1979.

‘R. E. Kirkup and L. R. Nelson, “City Fights Grease ml odor

problems in Sewer Systems,” Public Works Magazine, October

1977.
Ch. 7—The Use of Genetically Engineered Micro-Organisms in the Environment . 125

pounds of bacteria is unlikely to have any effect.

Thus far, the Environmental Protection Agency

(EPA) has not recommended adding bacteria to

municipal systems; however, EPA suggests that

they might be useful in smaller installations and

for specific problems in large systems.

Dry formulations are available for use in

cleaning drains and pipes in smaller installations, such as restaurants and other food processing facilities. In restaurants, the bacteria are

added to the drain at the end of the workday.

Bacteria have been selected for their inability to

produce hydrogen sulfide, which means that

the degrading process does not produce the unpleasant odors frequently encountered in the

digestion of oils and fats.6

As of November 1979, the pollution control

industry had few plans for the genetic manipulation of bacteria, except for the selection of

naturally occurring better performers. Consumer resistance to “mutants” is a factor that

discourages the move to microbial genetics.

Probably even more important is the high cost

of establishing and maintaining microbial genetics laboratories. It has been estimated that the

cost of carrying a single Ph. D. microbial geneticist is over $100,000 annually.7 This expense is

quite high relative to the $2 million to $4 million

sales of all biological pollution control companies in 1978.8

Resistance to the use of genetically manipulated bacteria is not universal. Many industrial

wastes are oxidized to nontoxic chemicals by

biological treatment in aerated lagoons. The

process depends on the presence of microbes in

the lagoons; over time, those that grow best on

the wastes come to dominate the microbial populations. Three companies now sell bacteria

that they claim outperform the indigenous

strains found in the lagoons. E.g., the Polybac

Corp. has sold its products to all seven Exxon

biological waste treatment plants to treat chemical wastes. One of its formulations has been

used to degrade toxic dioxins from an herbicide

spill. One month’s treatment with the bacterial

formulation reduced the orthochlorophenol

concentration from 600 to 25 ppm in a 20,000 -

gal lagoon.9

Sybron/Biochemical, a division of Sybron

Corp., sells cultures of bacteria that are intended to aid in the biological oxidation of industrial wastewater; this company also lists 20

different cultures for application to specific

wastes. Patent number 4,199,444 was granted

on April 22, 1980, for a process involving the

use of a mutant bacterial culture to decolor

waste water produced in Kraft paper processing.l0 Other patents are pending on a mixture of

two strains that degrade grease and a strain that

degrades “nonbiodegradable” detergents. 11

There is disagreement about the value of adding microbes to decontaminate soils or waters.

One point of view argues that serious spills frequently sterilize soils, and that adding microbes

is necessary for any biodegradation. The other

contends that encouraging indigenous microbes

is more likely to succeed because they are acclimated to the spill environment. Added bacteria have a difficult time competing with the

already-present microbial flora. In the case of

marine spills, bacteria, yeast, and fungi already

present in the water participate in degradation,

no one has been able to demonstrate the usefulness of added microbes.

Commercial applications—market size

and prospects

The estimated market size of pollution-control biological products in 1978 was $2 million to

$4 million, divided among some 20 companies,

‘Anon., Wean That Sewage System With Bugs!” Environmental

Science and 7’echno/ogy 13:1 198-1199, 1979.

7Anon., “Biotechnology DNA Research Expenditures in U.S. May

Reach $500 Million in 1980, With About $150-200 Million for Commercial Products, ” Hill told. Drug Research Reports, “The Blue

Sheet, ” May 28, 1980, p. 22.

aAnon., Business Week, July 5, 1976, p. 280; Chemical Week

121:47, 1977; and Food Engineering 49: 138, 1977, cited in T. Gassner, “Microorganisms for Waste Treatment, ” Microbial Technol-

~gv, 2 cd., vol. II, (London: Academic Press, 1979), pp. 211-222.

9See footnote 6.

IOL. Davis, J. E. Blair, and C, W. Randall, “Communicant ion:

Development of Color Removal Potential in Organisms Treating

Pulp and Paper Wastewater,” J. Water Pollution Control Fed., Februarv 1978, pp. 382-385.

i IF. Spraher and N. Tekeocgak, “Foam Control and Degradation

of Nonionic Detergent,” Industrial Wastes, January/February 1980;

L. David, J. E. Blair, and C. Randall, ‘(Mixed Bacterial Cultures Leak

‘Non-Biodegradable’ Detergent,” industrial Wastes, May/June

1979.
126 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

and the potential market was estimated to be as

much as $200 million.12 These estimates can be

compared to Polybac’s own sales records. In

1976, its first year, its sales totaled $0.5 million

and in 1977, $1.0 million. It expects to reach $5

million in 1981.

To date genetically engineered strains have

not been applied to pollution problems. At least

one prominent genetic engineering company

has decided not to enter the pollution control

field, concluding that it was improbable that

added microbes could compete with indigenous

organisms. More specifically, the possibility of

liability problems make the approach even less

attractive. Pollution control requires that “new”

life forms be released into the environment,

which is already seen as precariously balanced.

Such new forms might cause health, economic,

or environmental problems. The problems of

liability that might arise from such applications

are enough to deter entrepreneurs from contemplating work in the field at this time.

An additional reason for the reluctance of

some companies to engage in this activity is that

the opportunities for making money are limited.

Selling microbes, rather than their products,

may well be a one-shot opportunity. The microbes, once purchased, might be propagated

by the buyer. Nevertheless, at least two small

companies have announced that they are pursuing efforts to use genetic engineering.

The low-key efforts in this field might accelerate quickly if a significant breakthrough occurred, To date, no “new” organism has appeared that will degrade previously intractable

chemicals. The effect of such a development

might be enormous.

Genetic research in pollution control

The Oil and Hazardous Materials Spills

Branch of EPA currently supports research

aimed at isolating organisms to degrade three

specific chemical compounds. The work is being

carried out on contract; as of November 1979,

no field trials of the organisms had been under-

‘

zSee footnote 8.

taken. Two of the toxic chemicals, pentachorophenol and hexachlorocyclopentadiene, are

relatively long-lived compounds and present

long-term problems. A fungus and a bacterium

that can degrade the first compound have been

isolated,13 and Sybron/Biochemical already sells

a culture specifically for pentachlorophenol

degradation. The third toxic compound is methyl parathion. Its inclusion is more difficult to

understand, since it is degraded within a few

days after its application as a pesticide.

Efforts have been made to isolate bacteria

that can degrade (2,4-dichlorophenoxy) acetic

acid (2,4-D) and (2,4,5-trichlorophenoxy) acetic

acid (2,4,5-T), the components of Agent

Orange. 14 Strains of the bacterium Alcaligenes

paradoxus rapidly degrade 2,4-D, and the

genetic information for the degradation activity

has been located on a plasmid. The investigator

who found that strain, while optimistic about

the opportunities for isolating and transferring

other resistance genes, has been unable to find

a bacterium that degrades 2,4,5-T or its very

toxic contaminant, 2,3,7,8-tetrachlorodibenzopara-dioxin (TCDD or dioxin).

By far the best known research in this area is

that of Dr. Ananda M. Chakrabarty who engineered two strains of Pseudomonas, each of

which has the ability to degrade the four classes

of chemicals found in oil spills. Chakrabarty

began with four different strains of Pseudomonas. None of them presented a threat to

human health, and each could degrade one of

the four classes of chemicals. His research

showed that the genes controlling the degrading

activities were located on plasmids. Taking advantage of the relative ease of moving such

genes among bacteria, he produced two recombinant bacteria.

Chakrabarty has presented evidence that his

bacterium degrades complex petroleum mixtures such as crude oil or “Bunker C“ oil, and he

‘SN. K. Thuma, P. E. O’Neill, S. G. Brownlee, and R. S. Valentine,

“Laboratory Feasibility and Pilot Plant Studies: Novel Biodegradation Processes for the Ultimate Disposal of Spilled Hazardous

Materials, ” National Environment Research Center, U.S. Environmental Protection Agency, Cincinnati, Ohio, 1978.

MJ. M< pemberton, “pesticide Degrading plasmids: A Biological

Answer to Environmental Pollution by Phenoxyherbicides,” Arnbio

8:202-205, 1979.
Ch. 7—The Use of Genetically Engineered Micro-Organisms in the Environment ● 127

has proposed a method for using it to clean up

oil spills. The bacteria are to be grown in the

laboratory, mixed with straw, and dried. The

bacteria-coated straw can be stored until

needed, then dropped from a ship or aircraft

onto oil spills. The straw absorbs the oil and the

bacteria degrades it. 15 To completely cleanup a

spill will probably require mechanical efforts in

addition to the biological attack. It was the production of one of Chakrabarty’s strains that led

to the Supreme Court decision on “the patenting

of life. ” (See ch. 12 for further details.)

The essential difference between the wellpublicized Chakrabarty approach and a less

well-known one is that all the desired activities

in Chakrabarty’s approach are combined in a

single organism; while in the other method,

bacteria bearing single activities are mixed

together to yield a desired “formulation.” In yet

another approach, Sybron/Biochemical uses

mutation and selection to produce specialized

degradation activities. It also sells mixed

cultures for some applications.

The single-organism, multiple-enzyme system

has the advantage that every bacterium can attack a number of compounds. The mixed formulations allow the preferential proliferation of

bacteria that feed on the most abundant chemical; then, as that chemical is exhausted, other

bacteria, which flourish on the next most abundant chemical, become dominant. The preferential survival of only one or a few strains in a

mixed formulation might result in no bacteria

being available to degrade some compounds.

The multienzyme bacteria, on the other hand,

can degrade one chemical after another, or

alternatively, more than one at the same time.

Federal research support for

engineering microbes to detoxify

hazardous substances

EPA currently limits its support to research

aimed at selecting indigenous microbes, an area

lspa~ent specification I 436 573, Mav 19, 1976, Patent office,

London, England.

that has already attracted some commercial

research support. Commercial firms are looking

for large-scale markets, such as sewerage systems, or commonly occurring smaller markets,

such as gasoline spills and common industrial

wastes.

Whatever potential exists in identifying,

growing, and using naturally occurring microbes for pollution control pales beside the opportunities offered by engineering new ones,

Unfortunately, the potential risks increase as

well. EPA has taken a preliminary step toward

assessing the risks by soliciting studies to determine what environmental risks may exist from

accidentally or deliberately released engineered

microbes.

Summary

While some unreported efforts may be

underway, genetics has apparently been little

applied to pollution abatement. Nevertheless,

the production of “new” life forms that offer a

significant improvement in pollution control is a

possibility. The constraints are questions of

liability in the event of health, economic, or environmental damage; the contention that added

organisms are not likely to be a significant improvement; and the assumption that selling

microbes rather than products or processes is

not likely to be profitable.

The factors that have discouraged developments in this area would probably become less

deterring if convincing evidence were found

that microbes could remove or degrade an intractable pollutant. In the meantime, the research necessary to produce marked improvements has been inhibited. Overcoming this inhibition may require a governmental commitment to support the research, to buy the

microbes, and to provide for protection against

liability suits. Such a governmental role would

be in keeping with its commitment to protecting

health and the environment from the toxic effects of pollutants.
128 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Issue and Options—Biotechnology

ISSUE: How can the Federal Government promote advances in biotechnology and genetic engineering?

The United States is a leader in applying genetic engineering and biotechnology to industry. One reason is the long-standing commitment by the Federal Government to the funding

of basic biological research; several decades of

support for some of the most esoteric basic research has unexpectedly provided the foundation for a highly useful technology. A second is

the availability of venture capital, which has allowed the formation of small innovative companies that can build on the basic research.

The argument for Government promotion of

biotechnology and genetic engineering is that

Federal help is needed in those high priority

areas not being developed by industry.

The argument against such assistance is that

industry will develop everything of commercial

value without Federal help.

A look at what industry is now attempting indicates that sufficient investment capital is

available to pursue specific manufacturing objectives, such as for interferon and ethanol, but

that some high-risk areas that might be of interest to society, such as pollution control, may

need promotion by the Government. Other

areas, such as continued basic biological research, might not be profitable soon enough to

attract industry’s investment, Specialized education and training are areas in which the Government has already played a major role, although

industry has both supported university training

and conducted its own inhouse training.

OPTIONS:

A. Congress could allocate funds specifically for

genetic engineering and biotechnology R&D in

the budget of appropriate agencies, such as

the National Science Foundation (NSF), the

U.S. Department of Agriculture (USDA), the

Department of Health and Human Services

(DHHS), the Department of Energy (DOE), the

Department of Commerce (DOC), and the Department of Defense (DOD).

Congress has a long history of recognizing

areas of R&D that need priority treatment in

the allocation of funds. Biotechnology has not

been one of these. Even though agencies like

NSF receive congressional funding, its Alternative Biological Sources of Materials program

is one of the few applied programs that is not

congressionally mandated. As a result, the fiscal

year 1980 budget saw a reduction in the allocation of funds, from $4.1 million in 1979 to $2.9

million. A congressionally mandated program,

analogous to the successful NSF Earthquake

Hazard Mitigation program, could be written

into law. other programs, such as the competitive grants program at USDA (or the Office

of Basic Biological Research at DOE), are also

modestly funded.

Increasing the amount of money in an agency’s biotechnology program could bring criticism from other programs within each agency if

their levels of funding are not increased commensurately. The Competitive Grants Program

at USDA has similar problems; those who are

most critical of it argue that it should not take

funds from traditional programs. Nevertheless,

Congress could promote two types of programs:

those with long-range payoffs (basic research),

and those which industry is not willing to undertake but that might be in the national interest.

B. Congress could establish a separate Institute

of Biotechnology as a funding agency.

The merits of a separate institution lie in the

possibility of coordinating a wide range of efforts, all related to biotechnology. Among present organizations, biotechnology and applied genetics cut across several institutes and divisions

within them. Medically oriented research falls

primarily under the domain of the National Institutes of Health (NIH). EPA is concerned with

the prevention of pollution; while NSF’s effort in

biotechnology has been restricted to modest

support scattered through several divisions.
Ch. 7—The Use of Genetically Engineered Micro-Organisms in the Environment ● 129

The creation of an organization such as the National Technology Foundation (H.R. 6910) would

represent the kind of commitment to engineering, in general, that currently does not exist.

Competition for funds within other agencies

would be avoided, since funding would now occur at the level of congressional appropriations.

A separate institute, carrying the stamp of

Government recognition, would make it clear to

the public that this is a major new area with

great potential. This might foster greater academic and commercial interest in biotechnology

and genetic engineering.

On the other hand, biotechnology and genetic

engineering cover such a broad range of disciplines that a single agency would overlap the

mandates of existing agencies. Furthermore, the

creation of yet another agency carries with it all

the disadvantages of increased bureaucracy and

competition for funds at the agency level.

c. Congress could establish research centers in

universities to foster interdisciplinary approaches to biotechnology. In addition, a program of training grants could be offered to

train scientists in biological engineering.

The successful use of biological techniques in

industry depends on a multidisciplinary approach involving biochemists, geneticists, microbiologists, process engineers, and chemists.

Little is now being done, publicly or privately,

to develop expertise in this interdisciplinary

area.

In 1979, President Carter proposed the creation of generic technology centers (useful to a

broad range of industries) as one way to stimulate innovation. The centers would conduct

the kind of research that an individual company

might not consider cost effective, but that might

ultimately benefit several companies. Each center would be jointly funded by Government and

industry, with Government providing the seed

money and industry carrying most of the costs

within 5 years. If the centers were established

at universities, startup costs could be minimized.

Several congressional bills contain provisions

for centers similar to these. For example, on

October 21, 1980, President Carter signed into

law a bill (S. 1250) that would establish Centers

for Industrial Technology to foster research

links between industry and universities. They

would be affiliated with a university or nonprofit institution.

One or more of these centers could be specifically designated to specialize in biotechnology.

In addition, training grants could be used to

support the education of biotechnologists at the

centers or elsewhere. Currently, there is no nationwide training program to train students in

this discipline. Education programs, especially

for the postgraduate and graduate training of

engineers, could further the idea of using biological techniques to solve engineering problems.

D. Congress could use tax incentives to stimulate

biotechnology.

The tax laws could be used to stimulate biotechnology in several ways. First, they could expand the supply of capital for small high-risk

firms, which are generally considered more innovative than established firms, because of

their willingness to undertake the risks of innovation. Much of the pioneering work in the

industrial application of genetic techniques has

been done by such firms. By nature, they are

speculative, high-risk investments. Second, the

tax law could provide special subsidies to new

high-technology firms, which cannot use the

standard investment incentives, such as the investment tax credit, because they usually have

no taxable profits for the first several years

against which to apply the tax credit. Third, tax

incentives could be provided for both established and new firms to make the investment of

money for R&D more attractive.

There are a number of ways to expand the

supply of venture capital. One is to decrease the

tax rate on capital gains or the period an asset

must be held for it to be considered a capital

gain rather than ordinary income. This change

could be limited to stocks in high-technology

firms in order to focus its impact and minimize

revenue loss. Other options involving the stock

of high-technology companies are: a tax credit

to the investor who purchases the stock; defer-
130 ● Impacts of Applied Genetics —Micro-Organisms, Plants, and Animals

ment of capital gains taxes on the sales of these

stocks if the proceeds are reinvested into similarly qualifying stock; and more liberal capital

loss provisions.

In addition to focusing on the supply of capital, tax policy could attempt to directly increase

the profitability of potential growth companies.

Since most are not profitable for several years,

they cannot take full advantage of the investment tax credit—or even the provision for carrying net operating losses back 3 years and forward 7 years to offset otherwise taxable profits.

Two proposals may remedy this situation. First,

the investment tax credit could be refundable to

the extent it exceeded any tax liability of the

firm. A preliminary estimate of the revenue loss

for this proposal was $1 billion for 1979. Second, new companies could be permitted to

carry net operating losses forward for 10 years.

This change would give new firms the same

number of years over which to deduct losses as

established firms.

The final type of tax incentive is directed at

increasing R&D expenditures. Two major proposals would permit companies to take tax credits on a certain percentage of their R&D expenses, and on contributions to universities for

research.

The R&D credit has been advocated for several reasons. First, it would increase the aftertax return on R&D investments, making them

more attractive. Second, it would reduce the

degree of risk on such investments; with a 10-

percent credit, the real after-tax expense of a $1

million investment is $900,000. Finally, it would

give firms maximum flexibility in selecting projects for investment.

Questions have been raised about the cost effectiveness of the credit. For calendar year

1980, the Treasury Department estimated the

cost of a lo-percent R&D credit to be $1.9 million. Since R&D costs average only 10 to 20 percent of the total cost of bringing a new product

or process to the market, the net reduction in

the cost of commercializing an invention would

be 1 to 2 percent. Moreover, the commercial

stage of innovation is thought to be riskier and

costlier than the technical stage. Another problem is that the credit maybe a windfall for firms

that would be investing in R&D anyway. Finally,

the credit would subsidize R&D devoted to

minor product changes or incremental improvements in addition to R&D directed to more fundamental breakthroughs.

One of the provisions of a pending congressional bill (H.R. 5829) provides for a credit of 25

percent for incremental research expenditures

above those for a base period. By limiting the

credit to incremental expenditures, the bill

would create a more cost-effective credit, if

passed.

The final type of tax credit would be for corporate contributions to university research. The

Treasury Department estimated that a 25 percent credit for research in all fields would cost

$40 million in 1980. This credit would be targeted to more fundamental research and not to

the subsidy of short-term, incremental projects

that are usually a significant part of corporate

R&D budgets.

E. Congress could improve the conditions under

which U.S. companies can collaborate with

academic scientists and make use of the technology developed in universities in whole or in

part at the taxpayer expense.

Developments in genetic engineering have

kindled interest in this option. Nevertheless, the

Government’s role in fostering university-academic interaction is far from accepted. Such a

role may limit the flexibility of a cooperative effort. At the very least, disincentives such as patent restrictions could be removed.

The controversy has been summed up as follows.

At the next level of involvement, the Government could identify potential partners, and facilitate negotiations. A more active role would

involve the Government’s providing startup

funds. Finally, the Government could be a third

partner, sharing costs with industry and the

university. In this case, too large a Government

role could lead to Federal intervention in activities that should be the responsibility of business and industry.

‘Dennis Prager, G. S. omenn, Science 207: 379-384, 1980,
Ch. 7—The Use of Genetically Engineered Micro-Organisms in the Environment . 131

Certainly the Government can facilitate communication; in the health field, NIH, for instance, is an effective stimulus for contacts

among scientists.

The possible advantages and disadvantages of

university-industry interaction is illustrated by

a recent case involving a plan by Harvard University to collaborate with a genetic engineering

company. The plan had called for the establishment of a corporation to commercialize the

results of research being done in the laboratory

of a Harvard molecular biologist, who would

have been a principal in the firm. The University would not have been involved in financing

or managing the firm, which would also have

been housed separately from the campus. However, Harvard would have derived substantial

income if the company proved successful

through a gift of 10 to 15 percent of the equity

and a royalty on sales. After much debate

among the Harvard faculty and educators nationwide, the administration decided not to implement the plan because of concerns about

possible adverse impacts on academic values.

proponents of such arrangements argue that

the universities should reap some return from

the commercialization of research conducted

by their staff. In addition, many universities are

pressed for money, and joint ventures or research funding arrangements with industry

provide an attractive source of funds for research programs, especially when Federal support may decline. In return, industry would

gain access to the kind of fundamental research

that is the foundation for innovation and appears to be especially crucial in the field of genetic engineering, where the gap between basic

research and product development is smaller

than for other fields.

Opponents of these arrangements, especially

ones involving significant interaction as in the

Harvard plan, fear that the profit-seeking goals

of industry may be incompatible with academic

values. The following possible adverse impacts,

among others, have been articulated: 1) increase in secrecy, to the detriment of the free

exchange of ideas so important in academia; 2)

discrimination by the university in its hiring and

promotion policies in favor of those doing the

revenue-producing research; and 3) distortion

in the direction of research and in the training

of graduate students.

F. Congress could mandate support for specific

research tasks, such as pollution control using

microbes.

Investment in creating microbes to degrade

pollutants is slow because the potential market

is thought to be small and because of the severe

liability problems that might arise from intentional release of commercially supplied microbes.

But microbes may be useful in degrading intractable waste and pollutants. Genetic determinants for desired degradation activities may

be present in naturally occurring organisms, or

scientists may have to combine genes from different sources into a single organism. Current

research, however, is limited to isolating organisms from natural sources or from mutated

cultures. More elaborate efforts, involving recombinant DNA (rDNA) techniques or other

forms of microbial genetic exchange, will require additional effort.

A decision by the Federal Government to support research and to reduce liability concerns is

probably needed before the potential of microbial control of pollution can be realized. Federal

activity might depend on the results of an evaluation of the technical feasibility of microbial

pollution control, which could be made by

either an interagency task force or a special

commission. If the evaluation is negative, Congress might elect to do nothing to encourage the

technology. If the evaluation is positive, Congress might select from the following suboptions:

1.

2<

Initiate no research support nor any Federal relief from or limit on potential liability claims. This option would not foreclose

private commercial efforts, but it would

limit them because of restricted research

funds and large liability questions. If sufficiently large markets were anticipated or

found, the limitations would be overcome.

Initiate research support programs. Research might be directed at problems

posed by particular pollutants (contract re-
132 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

3.

4.

5.

6.

search). Federal support of biological research is managed by several agencies, and

this course would create few, if any, major

administrative problems.

Guarantee markets for particular products. In addition to patent protection,

which would be of little value in the case of

an organism purposefully disseminated

into the environment, the Government

could offer to buy desirable microbes. This

public sector market might provide enough

incentive to research to make Federal funding unnecessary, or the market incentive

and research support might be used jointly.

Fix a limit on liability and set up liability insurance, funded partly or wholly by the

Government. This option would reduce the

financial risk for entrepreneurs who venture to clean up pollutants with microbes,

Such an insurance scheme would require

that a Federal agency (EPA, for instance) be

satisfied that little risk was attendent in the

use of the microbe.

Arrange a scheme to test micro-organisms

for known and anticipated risks before

they are released. The Federal Government

might have to bear these costs as part of a

research program.

Leave most efforts to industry and allow

each Government agency to develop programs in the fields of genetic engineering

and biotechnology as it sees fit.

This option, currently the status quo, seems

to be favored by some industry officials. If it is

worth doing, they argue, industry will do it. To

a large extent, the availability of venture capital

in the United States has allowed many companies to pursue projects that are deemed practical and economically important. The production of interferon, insulin, ethanol, ethylene

glycol, and fructose are cited as examples of

successful applications that were motivated by

industry.

Generic research, or research that is fundamentally useful to a broad range of companies, will probably not be undertaken by any

one company. When the payoff does not come

soon enough, the Government has traditionally

taken the responsibility for funding the work.

E.g., NIH supported 717 basic research projects

involving rDNA in fiscal year 1980 at a cost of

$91.5 million. Similarly, high-risk research with

high capital costs would be likely targets for

Government support.

Leaving all R&D in industry’s hands would

still produce major commercial successes, but

would not ensure the development of generic

knowledge or the undertaking of high-risk projects.
Part II

Agriculture

Chapter 8—The Application of Genetics to Plants . . . . . . . . . . . . . . . . . . . . . . . . . . . , . . . . . . 137

Chapter 9—Advances in Reproductive Biology and Their Effects on Animal Improvement . . . 167
Chapters

The Application of

Genetics to Plants
Chapters

Page

Perspective on Plant Breeding. . . . . . . . . . . . . . . 137

The Plant Breeder’s Approach to

Commercialization of New Varieties . . . . . . . 138

Major Constraints on Crop Improvement. . . . . 139

Genetic Technologies as Breeding Tools . ......140

New Genetic Technologies for Plant Breeding .141

Phase I: Tissue Culture to Clone Plants ... , . 141

Phase H: Engineering Changes to Alter

Genetic Makeup; Selecting Desired Traits. . 144

Phase 111: Regenerating Whole Plants From

Cells in Tissue Culture . . . . . . . . . . . . . . . . 146

Constraints on the New Genetic Technologies . 149

Technical Constraints. . ................149

Institutional Constraints . . . . , . . . . . . , . . . . 150

Impacts on Generating New Varieties . . . . . . . . . 151

Examples of New Genetic Approaches . ......152

Selection of Plants for Metabolic Efficiency .. .152

Nitrogen Fixation. . . . . . . . . . . . . . . . . . . . . . . 152

Genetic Variability, Crop Vulnerability, and

Storage of Germplasm. . . . . . . . . . . . . . . . . . 154

The Amount of Genetic Erosion That Has

Taken Place . . . . . . . . . . . . . . . . . . . . . . . .. 154

The Amount of Germplasm Needed . ........154

The National Germplasm System . . . . . . . . . . . 155

The Basis for Genetic Uniformity . . . . . . . . . . . 157

Six Factors Affecting Adequate Management

of Genetic Resources. . . . . . . . . . . . . . . . . . . 158

Page

Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160

Issues and Options—Plants . . . . . . . . . . . . . . 161

Technical Notes . . . . . . . . . . . . . . . . . . . . . 162

Tables

Table No. Page

24. Average Yield per Acre of Major Crops in

1930 and 1975. , . . . . . . . . . . . . . . . . . . . . . 137

2.5. Some Plants Propagated Through Tissue

Culture for Production or Breeding. . . . . . . 143

26. Representative List of Tissue Culture

Programs of Commercial Significance in

the United States. . . . . . . , . . . . . . . . . . . . 143

27. Gene Resource Responsibilities of Federal

Agencies . . , . . . . . . . . . . . . . . . . . . . . . 155

28. Estimated Economic Rates of Return From

Germplasm Accessions . . . . . . . . . . . . . . 156

29. Acreage and Farm Value of Major U. S. Crops

and Extent to Which Small Numbers of

Varieties Dominate Crop Average . . . . . . 157

Figures

Figure No. Page

28. The Process of Plant Regeneration From

Single Cells in Culture . . . . . . . . . . . . . . . . 147

29. A Model for Genetic Engineering of Forest

Trees. . . . . . . . . . . . . . . . . . . . . . . . . . . 149
Chapter 8

The Application of Genetics to Plants

Perspective on plant breeding

As primitive people moved from hunting and

gathering to farming, they learned to identify

broad genetic traits, selecting and sowing seeds

from plants that grew faster, produced larger

fruit, or were more resistant to pests and diseases. Often, a single trait that appeared in one

plant as a result of a mutation (see Tech. Note 1,

p. 162.) was selected and bred to increase the

trait’s frequency in the total crop population.

Mendel’s laws of trait segregation enabled

breeders to predict the outcomes of hybridization and refinements in breeding methods. (See

app. II-A.) Consequently, they achieved breeding objectives faster and with more precision,

significantly increasing production. During the

past 80 years classical applied genetics has been

responsible for:

increased yields;

overcoming natural breeding barriers;

increased genetic diversity for specific

uses;

expanded geographical limits where crops

can be grown; and

improved plant quality.

Since the beginning of the 20th century, plant

breeders have helped increase the productivity

(see Tech. Note 2, p. 162.) of many important

crops for food, feed, fiber, and pharmaceuticals

by successfully developing cultivars (cultivated

varieties) to fit specific environments and production practices. Some breeding objectives

have met the needs of the local farmer, while

other genetic improvements have been applied

worldwide. The commercial development of

hybrid corn in the 1920’s and 1930’s and of

“green revolution” wheats in the 1950’s and

1960’s are but two examples of how plant

breeding has affected the supply of food available to the world market. (See Tech. Note 3, p.

162.) A comparison of average yields per acre in

1930 and 1975 in table 24 gives a measure of the

contribution of genetics.1

It is impossible to determine exactly to what

degree applied genetics has directly contributed

to increases in yield, because there have been

simultaneous improvements in farm management, pest control, and cropping techniques

using herbicides, irrigation, and fertilizers. Various estimates, however, indicate that applied

genetics has accounted for as much as 50 percent of harvest increases in this century. The

yield superiority of new varieties has been a major impetus to their adoption by farmers. Historically, the primary breeding objective has been

to maintain and improve crop yields. Other

‘C,. F. Sprague, D. E. Alexander, and J. W. Dudley, “Plant Breeding and Genetic Engineering: A Perspective, ” Bioscience SO(I): 17,

1980.

Table 24.—Average Yield per Acre of Major Crops

in 1930 and 1975

Average yield per acre Percent

1930 1975 Unit increase

Wheat. . . . . . . . . . . . . 14.2

Rye . . . . . . . . . . . . . . . 12.4

Rice. . . . . . . . . . . . . . . 46.5

Corn . . . . . . . . . . . . . . 20.5

Oats . . . . . . . . . . . . . . 32.0

Barley. . . . . . . . . . . . . 23.8

Grain sorghum. . . . . . 10.7

Cotton . . . . . . . . . . . . 157.1

Sugar beets . . . . . . . . 11.9

Sugarcane . . . . . . . . . 15.5

Tobacco. . . . . . . . . . . 775.9

Peanuts . . . . . . . . . . . 649.9

Soybeans. . . . . . . . . . 13.4

Snap beans . . . . . . . . 27.9

Potatoes. . . . . . . . . . . 61.0

Onions . . . . . . . . . . . . 159.0

Tomatoes:

Fresh market. . . . . 61.0

Processing. . . . . . . 4.3

Hops. . . . . . . . . . . . . . 1,202.0

30.6

22.0

101.0

86.2

48.1

44.0

49.0

453.0

19.3

37.4

2,011.0

2,565.0

28.4

37.0

251.0

306.0

166.0

22.1

1,742.0

Bushels

Bushels

Bushels

Bushels

Bushels

Bushels

Bushels

Pounds

Tons

Tons

Pounds

Pounds

Bushels

Cwt

Cwt

Cwt

Cwt

Tons

Pounds

115

77

117

320

188

62

141

159

295

112

33

129

92

172

413

45

SOURCE: U.S. Department of Agriculture, P/ant Genet~c Resources.. Cozwewaflon and Use (Washington, D. C.: USDA, 1979).

137
138 ● Impacts Applied Genetics—Micro-Organisms, Plants, and Animals

breeding objectives are specific responses to the

needs of local growers, to consumer demands,

and to the requirements of the food processing

firms and marketing systems.

Developing new varieties does the farmer little good unless they can be integrated profitably

into the farming system either by increasing

yields and the quality of crops or by keeping

costs down. The three major goals of crop

breeding are often interrelated. They are:

●

●

●

to maintain or increase yields by selecting

varieties for:

—pest (disease) resistance;

—drought resistance;

—increased response to fertilizers; and

—tolerance to adverse soil conditions.

to increase the value of the yield by selecting varieties with such traits as:

—increased oil content;

—improved storage qualities;

—improved milling and baking qualities;

and

—increased nutritional value, such as higher levels of proteins.

to reduce production costs by selecting

varieties that:

—can be mechanically harvested, reducing

labor requirements;

—require fewer chemical protestants or

fertilizers; and

-can be used with minimum tillage systems, conserving fuel or labor by reducing the number of cultivation operations.

The plant breeder’s approach to

commercialization of new varieties

The commercialization of new varieties

strongly depends on the genetic variability that

can be selected and evaluated. A typical plant

breeding system consists of six basic steps:

1. Selecting the crop to be bred.

2. Identifying the breeding goal.

3. choosing the methodological approach

needed to reach that goal.

4. Exchanging genetic material by breeding.

5. Evaluating the resulting strain under field

conditions, and correcting any deficiencies

in meeting the breeding goal.

6. Producing the seed for distribution to the

farmer.

The responsibilities for the different breeding

phases are distributed but interactive. In the

United States, responsibility for crop improvement through plant breeding is shared by the

Federal and State governments, commercial

firms, and foundations,2 Although some specific

genes have been identified for breeding programs, most improvements are due to gradual

selection for favorable combinations of genes in

superior lines. The ability to select promising

lines is often more of an art (involving years of

experience and intuition) than a science.

The plant breeder’s approach is determined

for the most part by the particular biological

characteristics of the crop being bred—e.g., the

breeder may choose to use a system of inbreeding or outbreeding, or the two in combination,

as an approach to controlling and manipulating

genetic variability. The choice is influenced by

whether a particular plant in question naturally

fertilizes itself or is fertilized by a neighboring

plant. To a lesser degree, the breeding objectives influence the choice of methods and the sequence of breeding procedures.

Repeated cycles of self-fertilization reduce

the heterozygosity in a plant, so that after numerous generations, the breeder has homozygous, pure lines that breed true. (See Tech. Note

4, p. 162.) Cross-fertilization, on the other hand,

results in a new mixture of genes or increased

genetic variability. Using these two approaches

in combination produces a hybrid—several lines

are inbred for homozygosity and then crossed

to produce a parental line of enhanced genetic

potential. More vigorous hybrids can be selected for further testing. The effects of hybrid

vigor vary and include earlier germination, increased growth rate or size, and greater crop

uniformity.

A second method for exchanging or adding

genes is achieved through altering the number

of chromosomes, or ploidy (see Tech. Note 5, p.

162.), of the plant. Since chromosomes are

‘National Academy of Sciences, Conservation of Germplasm Resources: An Imperative, Washington, D. C., 1978.
Ch. 8—The Application of Genetics to Plants ● 139

generally inherited in sets, plants whose ploidy

is increased usually gain full sets of new

chromosomes. over one-third of domesticated

species are polyploids. 3 Generally, crop improvement due to increased ploidy corresponds

to an overall enlargement in plant size; leaves

can be broader and thicker with larger flowers,

fruits, or seeds. A well-known example is the

cultivated strawberry, which has four times

more chromosomes than the wild type, and is

much fleshier.

Another technique, called backcrossing can

improve a commercially superior variety by lifting one or more desirable traits from an inferior

one. Generally, this is accomplished by making a

series of crosses from the inferior to the superior plant while selecting for the desired traits in

each successive generation. Self-fertilizing the

last backcrossed generation results in some

progeny that are homozygous for the genes being transferred and that are identical with the

superior variety in all other respects. Single

gene resistance to plant pests and disease-causing agents has been successfully transferred

through backcrossing.

Major constraints on crop

improvement

Two of the many constraints on crop breeding are related to genetics.

Many important traits are determined by several

genes.

The genetic bases for improvements in yield

and other characteristics are not completely

defined, mainly because most biological traits,

such as plant height, are caused by the interaction of numerous genes. Although many—perhaps thousands—of genes contribute to quantitative traits, much variation can be explained

by a few genes that have major impact on

the observable appearance (phenotype) 4—e.g.,

the height of some genetic dwarves in wheat

can be doubled by a single gene. Many other

genes contribute to the general health of the

3W. J. C. Lawrence, Plant Breeding [London: Edward Arnold Ltd.,

1968).

4J. N. Thompson, Jr., “Analysis of Gene Number and Development in Polygenic Systems, ” Stadler Genetics Symposium 9:63.

plant (such as resistance to pests and diseases),

although some of their contributions are small

and difficult to assess. Favorable combinations

of genes result in plants well-adapted to particular growing conditions and agronomic practices. With thousands of genes in a single plant

contributing to overall fitness, the possible combinations are almost infinite.

Most poor combinations of genes are eliminated by selection of the best progeny; initially

favorable combinations are preserved and improved. Literally millions of plants may be examined each year to find particularly favorable

genotypes for development into new breeding

stocks. Increasingly sophisticated field testing

procedures, as well as advanced statistical analyses, are now used to evaluate the success of

breeding efforts. Overall yield is still the most

important criterion for success, although considerable care is taken to test stress tolerance,

pest and disease resistances, mechanical harvestability, and consumer acceptability. Breeding programs with specialized goals often use

rapid and accurate chemical procedures to

screen lines and progeny for improvements.

Because the vigor of the plant depends on the

interaction of many genes, it has been difficult

to identify individual genes of physiological

significance in whole plants. As a result, many

important genes have not been mapped in

major crop species. There is little doubt that

breeders would select traits like photosynthetic

efficiency (the ability to convert light to such

organic compounds as carbohydrates) or mineral uptake if the genes could be identified and

manipulated in the same ways that resistance is

selected for pathogens.

It is uncertain how much genetic variation for improvement exists.

Although the world’s germplasm resources

have not been completely exploited, it has

become more difficult for breeders to improve

many of the highly developed varieties now in

use-e. g., height reduction in wheat has made

enormous contributions to its productivity, but

further improvement on this basis seems to be

limited.5 A parallel condition in the potato crop

‘N. ~. Jensen, “Limits to Growth in World Food Product ion,” Science 201:317, 1978.
140 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Photo credit: U.S. Department of Agriculture

Bundles of wheat showing variance in height

was recognized by the National Research Council’s Committee on Genetic Vulnerability of Major Crops:6

If we bear in mind the fairly recent origin of

modern potato varieties and that they are, for

the most part, derived from the survivors of the

late blight epidemics of the 1840’s in Europe and

North America, it seems likely that the genetic

‘Niitbnal Academy of Sciences, Genefic Vulnerability of Major

Crops, Washington, D.[I., 1972.

base was already somewhat narrow by the time

modern potato breeding got under way. The

five-fold increase in yield resulting from selection during the last 100 years of potato improvement has produced a group of varieties that are

genetically similar and unlikely to respond to

further selection for yield. In the long run response to selection for other characteristics is

also likely to be limited.

As these examples indicate, the level of genetic

homogeneity of some crops may make selection

for higher yields in general more difficult.

Nevertheless, while the genetic basis for overall

crop improvement is poorly understood, refinements in plant breeding techniques may increase the potential for greater efficiency in the

transfer of genetic information for more precise

selection methods, and as a new source of genetic variation.

Besides these two constraints, other pressures and limitations may also affect crop productivity; some are biological (see Tech. Note 6,

p. 162.), requiring technological breakthroughs,

while others are related to environmental,

social, and political factors. (See Tech. Note 7, p.

162.)—e.g., it has been argued that the agricultural rate of growth is declining: In 1976, the

U.S. Department of Agriculture (USDA) estimated that the total-factor productivity of U.S.

agriculture increased by 2 percent per year

from 1939 to 1960, but by only 0.9 percent from

the period of 1960 to 1970.7

7

11. S. Department of Agriculture, Economics, Statistics, and Cooperation Services, Agricultural Productivi?,v: EKpanding the Limits,

Agriculture Information Bulletin 431, Washington, D. C., 1979.

Genetic technologies as breeding tools

The new technologies may provide potentially useful tools, but they must be used in combination with classical plant breeding techniques to be effective. The technologies developed for classical plant breeding and those of

the new genetics are not mutually exclusive,

they are both tools for effectively manipulating

genetic information through methods that have

been adapted from genetic recombination observed in nature. Plant breeders have many

techniques for artificially controlling pollination—some are capable of overcoming natural

barriers such as incompatibility, Yet even

though one new technology-protoplasm fusion

—allows breeders to overcome incompatibility,

the new plant must still be selected, regenerated from single-cell culture, and evaluated

under field conditions to ensure that the genetic

change is stable and the attributes of the new

variety meet commercial requirements. Evacuation is still the most expensive and time-consuming step.
Ch. 8—The Application of Genetics to Plants . 141

New genetic technologies for

plant breeding

The recent breakthroughs in genetic engineering permit the plant breeder to bypass the

various natural breeding barriers that have

limited control of the transfer of genetic information. While the new technologies do not

necessarily offer the plant breeder the radical

changes that recombinant DNA (rDNA) technology provides the microbiologist, they will, in

theory, speedup and perfect the process of genetic refinement.

The new technologies fall into two categories: those involving genetic transformations

through cell fusion, and those involving the insertion or modification of genetic information

through the cloning (exactly copying) of DNA

and DNA vectors (transfer DNA). Most genetic

transformations require that enzymes digest

the plant’s impermeable cell wall, a process that

leaves behind a cell without a wall, or a protoplasm. Protoplasts can fuse with each other, as

well as with other components of cells. In

theory, their ability to do this permits a wider

exchange of genetic information.

The approach exploiting the new technologies is usually a three-phase program.

.

Phase 1. Isolated cells from a plant are established in tissue culture and kept alive.

Phase 11. Genetic changes are engineered in

those cells to alter the genetic makeup

of the plant; and desired traits are

selected at this stage, if possible.

Phase 111. The regeneration of the altered single

cells is initiated so that they grow into

entire plants.

This approach contains similarities to the genetic manipulation of micro-organisms. However,

there is one major conceptual difference. In

micro-organisms, the changes made on the cellular level are the goals of the manipulation.

With crops, changes made on the cellular level

are meaningless unless they can be reproduced

in the entire plant. Therefore, unless single cells

in culture can be grown into mature plants that

have the new, desired characteristics—a procedure which, at this time, has had limited success—the benefits of genetic engineering will

not be widespread. If the barriers can be overcome, the new technologies will offer a new

way to control and direct the genetic characteristics of plants.

PHASE 1: TISSUE CULTURE TO CLONE PLANTS

Tissue culture involves growing cells from a

plant in a culture or medium that will support

them and keep them viable. It can be started at

three different levels of biological organization:

with plant organs (functional units such as

leaves or roots);* with tissues (functioning aggregates of one type of cell, such as epidermal

cells (outermost layer) in a leaf; and with single

cells. Tissue cultures by themselves offer specific benefits to plant breeders; just as fermentation is crucial to microbial genetic technologies,

tissue culture is basic to the application of the

other new genetic technologies for plants.

The idea of growing cells from higher plants

or animals and then regenerating entire plants

from these laboratory-grown cells is not new.

However, a better scientific understanding now

exists of what is needed to keep the plant parts

alive.

In tissue culture, isolated single plant cells are

typically induced to undergo repeated cell divisions in a broth or gel, the resulting amorphous

cell clump is known as a callus. If culture conditions are readjusted when the callus appears, its

cells can undergo further proliferation. As the

resulting cells differentiate (become specialized), they can grow into the well-organized

tissues and organs of a complete normal plant.

The callus can be further subcultured, allowing

mass propagation of a desired plant.

At this time, it is not uncommon to produce as

many as a thousand plants from each gram of

starting cells; 1 g of starting carrot callus routinely produces 500 plants. The ultimate goal of

tissue culturing is to have these plantlets placed

in regular soil so that they can grow and develop into fully functional mature plants. The complete cycle (from plant to cell to plant) permits

production of plants on a far more massive

scale, and in a far shorter period, than is possible by conventional means. (See table 25 for a

*Also referred to as organ culture.
142 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

First stage in plant tissue culturing: inoculation

of plant tissue

cies basis. However, several commercial uses of

tissue culture already exist. (See table 26.)

Storage of Germplasm.—Tissue culture

can be used in the long-term storage of specialized germplasm, which involves freezing cells

and types of shoots. The culture provides stable

genetic material, reduces storage space, and

decreases maintenance costs.

Shows the gradual development of the plant t issue

on an agar medium

list of some p

culture. )

ants propagated through tissue

Each of the four stages of the complete

cycle—establishment in culture, organogenesis,

plantlet amplification, and reestablishment in

soil—requires precise biological environments

that have to be determined on a species-by-speCarrot tissues have been frozen in liquid nitrogen, thawed 2 years later, and regenerated

into normal plants. This technique has also

proved successful with morning glories, sycamores, potatoes, and carnations. Generally, the

technique is most useful for plant material that

is vegetatively propagated, although if it can be

generally applied it could become important for

other agriculturally important crops.

Production of pharmaceuticals and

Other Chemicals From Plant Cells.—Because plant cells in culture are similar to microorganisms in fermentation systems, they can be

engineered to work as “factories” to produce
Ch. 8—The Application of Genetics to Plants . 143

Table 25.—Some Plants Propagated Through

Tissue Culture for Production or Breeding

Agriculture and Poinsettia

horticulture Weeping fig

Vegetable crops Rubber plant

Asparagus Flowers

Beet African violet

Brussels sprouts Anthruium

Cauliflower Chrysanthemum

Eggplant Gerbera daisy

Onion Gloxinia

Spinach Petunia

Sweet potato Rose

Tomato Orchid

Fruit and nut trees Ferns

Almond Australian tree fern

Apple Boston fern

Banana Maidenhair fern

Coffee Rabbitsfoot fern

Date Staghorn fern

Grapefruit Sword fern

Lemon Bulbs

Olive Lily

Orange Daylily

Peach Easter lily

Fruit and berries Hyacinth

Blackberry

Grape

Pharmaceutical

Atropa

Pineapple

Strawberry Ginseng

Pyrethium

Foliage

Silver vase Silviculture (forestry)

Begonia Douglas fir

Cryptanthus Pine

Dieffenbachia Quaking aspen

Dracaena Redwood

Fiddleleaf Rubber tree

SOURCE: Office of Technology Assessment.

Table 26.—Representative List of Tissue Culture Programs of Commercial Significance in the United States

Industry Application Economic benefits

Asparagus industry. . . . . . . . . .

Chemical and pharmaceutical.

Citrus industry. . . . . . . . . . . . . .

Coffee industry . . . . . . . . . . . . .

Land reclamation. . . . . . . . . . . .

Ornamental horticulture. . . . . .

Pineapple industry . . . . . . . . . .

Strawberry industry. . . . . . . . . .

Rapid multiplication of seed stock Improved productivity, earliness, and spear quality

Biosynthesis of chemicals Reduced production costs

Propagation of medicinal plants High volumes of plants for planting

Virus elimination Improved quality, high productivity

Disease resistance breeding Disease resistance

Mass propagation Availability of select clones of wild species for revegetation

Mass propagation Reduced costs of certain species

Virus elimination of certain species

Introduction of new selections

Increased volumes of difficult selections

Mass propagation Improved quality in higher volumes

Mass propagation Rapid introduction of new strains

SOURCE: Office of Technology Assessment.
144 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

plant products or byproducts. In recent years,

economic benefits have been achieved from the

production of plant constituents through cell

culture. Among those currently produced commercially are camptothecin (an alkaloid with

antitumor and antileukemic activity), proteinase

inhibitors (such as heparin), and antiviral substances. Flavorings, oils, other medicinal, and

insecticides will also probably be extracted from

the cells.

The vinca alkaloids—vincristine and vinblastine, for instance—are major chemotherapeutic agents in the treatment of leukemias and

lymphomas. They are derived from the leaves

of the Madagascar periwinkle (Catharanthus

roseus). Over 2,000 kilograms (kg) of leaves are

required for the production of every gram of

vinca alkaloid at a cost of about $250/g. Plant

cells have recently been isolated from the periwinkle, immobilized, and placed in culture. This

culture of cells not only continues to synthesize

alkaloids at high rates, but even secretes the material directly into the culture medium instead

of accumulating it within the cell, thus removing the need for extensive extraction procedures.

Similarly, cells from the Cowage velvetbean

are currently being cultured in Japan as a

source of L-Dopa, an important drug in the

treatment of Parkinson’s disease. Cells from the

opium poppy synthesize both the plant’s normal

alkaloids in culture and, apparently, some alkaloids that have not as yet been found in extracts

from the whole plant.

Another pharmaceutical, diosgenein, is the

major raw material for the production of corticosteroids and sex steroids like the estrogens

and progestins used in birth control pill. The

large tuberous roots of its plant source,

Dioscorea, are still collected for this purpose in

the jungles of Central America, but its cells have

been cultured in the laboratory.

Other plant products, from flavorings and

oils to insecticides, industrial organic chemicals,

and sweeteners, are also beginning to be derived from plants in cell-cultures. Glycyrrhiza,

the nonnutritive sweetener of licorice, has been

produced in cultures of Glycyrrhiza glabra, and

anthraquinones, which are used as dye bases,

accumulate in copious amounts over several

weeks in cultures of the mulberry, Morinda citrifolia.

PHASE II: ENGINEERING CHANGES TO ALTER

GENETIC MAKEUP; SELECTING DESIRED TRAITS

The second phase of the cycle involves the

genetic manipulation of cells in tissue culture,

followed by the selection of desired traits,

Tissue culturing, in combination with the new

genetic tools, could allow the insertion of new

genetic information directly into plant cells.

Several approaches to exchanging genetic information through new engineering technologies

exist:

● culturing plant sex cells and embryos;

● protoplasm fusion; and

● transfer by DNA clones and foreign

tors.

These are then followed by:

. screening for desired traits.

vecCulturing Plant Cells and Embryos.–

Culturing the plant’s sex cells—the egg from the

ovary and the pollen from the anther (pollensecreting organ)—can increase the efficiency of

creating pure plant lines for breeding. Since sex

cells contain only a single set of unpaired

chromosomes per cell, plantlets derived from

them also contain only a single set. Thus, any

genetic change will become apparent in the regenerated plant, because a second paired gene

cannot mask its effect. Large numbers of haploid plants (cells contain half the normal number of chromosomes) have been produced for

more than 20 species. Simple treatment with

the chemical, colchicine, can usually induce

them to duplicate their genomes (haploid set of

chromosomes) —resulting in fully normal, diploid plants. The only major crop that has been

bred by this technique is the asparagus.8

If the remaining technical barriers can be

overcome, the technique can be used to enhance the selection of elite trees and to create

hybrids of important crops. Although still

‘J. G. Torreey, “Cytodifferentiation in Cultured Cells and Tissues, ”

l-/ortScience 12(2):138, 1977.
Ch. 8—The Application of Genetics to Plants . 145

primarily experimental, successful plant sex-cell

cultures have been achieved for a variety of

important cultivars, including rice, tobacco,

wheat, barley, oats, sorghum, and tomato. However, because the technique can lead to bizarre

unstable chromosomal arrangements, it has had

few applications.

Embryo cultures have been used to germinate, in vitro, those embryos that might not

otherwise survive because of basic incompatibilities, especially when plants from different

genera are crossed. Embryos may function as

starting material in tissue culture systems requiring juvenile material. They are being used

to speed up germination in such species as oil

palms, which take up to 2 years to germinate

under natural conditions.

Protoplasm Fusion.—In protoplasm fusion,

either two entire protoplasts are brought together, or a single protoplasm is joined to cell

components—or organelles—from a second protoplasm. When the components are mixed under

the right conditions, they fuse to form a single

hybrid cell. The hybrids can be induced to proliferate and to regenerate cell walls. The functional plant cell that results may often be

cultured further and regenerated into an entire

plant—one that contains a combination of genetic material from both starting plant cell progenitors. When protoplasts are induced to fuse, they

can, in theory, exchange genetic information

without the restriction of natural breeding barriers. At present, protoplasm fusion still has

many limitations, mainly due to the instability of

chromosome pairing.

Organelles are small, specialized components

within the cell, such as chloroplasts and mitochondria. Some organelles, called plastids, carry

their own autonomously replicating genes, as a

result, they may hold promise for gene transfer

and for carrying new genetic information into

protoplasts in cultures, or possibly for influencing the functions of genes in the cell nucleus.

(See Tech. Note 8, p. 163.)

The feasibility of protoplasm fusion has been

borne out in recent work with tobacco–a plant

that seems particularly amenable to manipulation in culture. An albino mutant of Nicotiana

tabacum was fused with a variety of a sexually

incompatible Nicotiana species. The resultant

hybrids were easily recognized by their intermediate light green color. They have now been

regenerated into adult plants, and are currently

being used as a promising source of hornworm

resistance in tobacco plants.

Transfer by DNA Clones and Foreign

Vectors.—Recombinant DNA technology

makes possible the selection and production of

more copies (amplification) of specific DNA

segments. Several basic approaches exist. In the

“shotgun” approach, the whole plant genome is

cut by one or more of the commercially available restriction enzymes. The DNA to be transferred is then attached to a plasmid or phage,

which carries genetic information into the plant

cell.—E.g., a gene coding for a protein (zein) that

is a major component of corn seeds has been

spliced into plasmids and cloned in micro-organisms. It is hoped that the zein-gene sequence

can be modified through this approach to increase the nutritional quality of corn protein

before it is reintroduced into the corn plant.

Foreign vectors are nonplant materials (viruses and bacterial plasmids) that can be used to

transfer DNA into higher plant cells. Transformation through foreign vectors might improve plant varieties or, by amplifying the desired DNA sequence, make it easier to recover a

cell product from culture. In addition, methods

have been discovered that eliminate the foreign

DNA from the transformed mixture, leaving

only the desired gene in the transformed plant.

The most promising vector so far seems to be

the tumor-inducing (Ti) plasmid carried by

Agrobacterium tumefaciens. This bacterium

causes tumorous growths around the root

crowns of plants. It infects one major group of

plants–the dicots (such as peas and beans), socalled because their germinating seeds initially

sprout double leaves. Its virulence is due to the

Ti plasmid, which, when it is transferred to

plant cells, induces tumors. Once inside the cell,

a smaller segment of the Ti plasmid, called TDNA, is actually incorporated into the recipient

plant cell’s chromosomes. It is carried in this

form, replicating right along with the rest of the
146 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

chromosomal DNA as plant cell proliferation

proceeds. Researchers have been wondering

whether new genetic material for plant improvement can be inserted into the T-DNA

region and carried into plant cell chromosomes

in functional form.

Adding foreign genetic material to the T-DNA

region has proved successful in several experiments. Furthermore, it has been found that

one type of plant tumor cell that contains

mutagenized T-DNA can be regenerated into a

complete plant. This new discovery supports

the use of the Agrobacterium system as a model

for the introduction of foreign genes into the

single cells of higher plants.

Many unanswered questions remain before

Agrobacterium becomes a useful vector for

plant breeding. Considerable controversy exists

about exactly where the Ti plasmid integrates

into the host plant chromosomes; some insertions might disrupt plant genes required for

growth. In addition, these transformations may

not be genetically stable in recipient plants;

there is evidence that the progeny of Ti-plasmidcontaining plants do not retain copies of the Ti

sequence. Finally, Agrobacterium does not readily infect monocots (a second group of plants),

which limits its use for major grain crops.

Another promising vector is the cauliflower

mosaic virus (CaMV). Since none of the known

plant DNA viruses has ever been found in plant

nuclear DNA, CaMV may be used as a vector for

introducing genetic information into plant

cytoplasm. Although studies of the structural

organization, transcription, and translation of

the CaMV are being undertaken, information

available today suggests that the system needs

further evaluation before it can be considered

an alternative to the Agrobacterium system.

Although work remains to be done on Tiplasmid and CaMV genetic mechanisms, these

systems have enormous potential. Most immediately, they offer ways of examining basic mechanisms of differentiation and genetic regulation

and of delineating the organization of the

genome within the higher plant cell. If this can

be accomplished, the systems may provide a

way of incorporating complex genetic traits into

whole plants in stable and lasting form.

Screening for Desired Traits.—The benefits of any genetic alteration will be realized

only if they are combined with an adequate system of selection to recover the desired traits. In

some cases, selection pressures can be useful in

recovery. g The toxin from plant pathogens, for

example, can help to identify disease resistance

in plants by killing those that are not resistant.

So far, this method has been limited to identifying toxins excreted by bacteria or fungi and

their analog; after sugarcane calluses were exposed to toxins of leaf blight, the resistant lines

that survived were then used to develop new

commercial varieties. In theory, however, it is

possible to select for many important traits.

Tissue culture breeding for resistance to salts,

herbicides, high or low temperatures, drought,

and new varieties that are more responsive to

fertilizers is currently under study.

Five basic problems must be overcome before

any selected trait can be considered beneficial

(see figure 28):

the trait itself must be identified;

a selection scheme must be found to identify cells with altered properties;

the properties must prove to be due to genetic changes;

cells with altered properties must confer

similar properties on the whole plant; and

the alteration must not adversely affect

such commercially important characteristics as yield.

While initial screens involving are easier to

carry out than screening tests involving entire

plants, tolerance at the cellular level must be

confirmed by inoculations of the mature plants

with the actual pathogen under field conditions.

PHASE III: REGENERATING WHOLE PLANTS

FROM CELLS IN TISSUE CULTURE

New methods are being developed to:

● increase the speed with which crops are

multiplied through mass propagation, and

. create and maintain disease-free plants.

Mass Propagation.—The greatest single

use of tissue culture systems to date has been

for mass propagation, to establish selected

‘J. F. Shepard, D. Bidney, and E. Shahin, “potato Protop]asts in

Crop Improvement,” Science 208:17, 1980.
Ch. 8—The Application of Genetics to Plants ● 147

Figure 28.—The Process of Plant Regeneration From Single Cells in Culture

Virus-free

Field performance tests

Cell multiplication

Tissue

Cell wall

removal

+ Root-promoting

hormones

The process of plant propagation from single ceils in culture can produce plants with selected characteristics. These selections must be tested in the field to evaluate their performance.

SOURCE: Office of Technology Assessment.

Photo credit: Plant Resources Institute

Multiplying shoots of jojoba plant in tissue culture on a

petri dish. These plants may potentiality be selected for

higher oil content

culture because of the increased speed with

sources of improved seed or cutting material.

(See table 26.) In some cases, producing plants

by other means is simply not economically competitive. A classic example is the Boston fern,

which, while it is easy to propagate from runner

tips, is commercially propagated through tissue

which it multiplies and the reduced costs of

stock plant maintenance. A tissue culture stock

of only 2 square feet (ft2

) can produce 20,000

plants per month. 10

Currently, mass production of such cultivars

as strawberries (see Tech. Note 9, p. 163.),

asparagus, oil palms, and pineapples is being

carried out through plant tissue cultures. 11 Very

recently, alfalfa was propagated in the same

way, giving rise to over 200,000 plants, several

thousand of which are currently being tested in

field trials. Also, 1,300 oil palms, selected for

high yield and disease resistance, are being

tested in Malaysia. 12 Other crops not produced

by this method but for which cell culture is an

important source of breeding variation include

‘OD. P. Holdgate, ‘(Propagation of Ornamental by Tissue Culture, ” in Plant Cell, Tissue, and Organ C’ulture, .1. Feinert and Y. P. S.

Bajaj (eds.) (New’ York: Sprin@r-\rerla~, 1977).

“T. Murashige, “Current Status of Plant Cell and organ Cultures, ” FfortScience )2(2):127, 1977.

‘

z

’’ The Second Green Revolution, ” special report, Business Week,

/W~. 25, 1980.
148 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

beets, brussels sprout, cauliflower, tomatoes,

citrus fruits, and bananas. Various horticultural

plants—such as chrysanthemums, carnations,

African violets, foliage plants, and ferns—are

also being produced by in vitro techniques.

Accelerating propagation and selection in

culture is especially compelling for economically important forest species for which traditional

breeding approaches take a century or more.

Trees that reach maturity within 5 years require approximately 50 years to achieve a useful

homozygous strain for further breeding. Species such as the sequoia, which do not flower

until they are 15 to 20 years old, require between 1 and 2 centuries before traits are stabilized and preliminary field trials are evaluated. Thus, tissue culture production of trees

has become an area of considerable interest.

Already, 2,500 tissue-cultured redwoods have

been grown under field conditions for comparison with regular, sexually produced seedlings.

(See app. II-B.) Loblolly pine and Douglas fir are

also being cultured; the number of trees that

can be grown from cells in 100 liters (]) of media

in 3 months are enough to reforest roughly

120,000 acres of land at a 12 x 12 ft spacing.13 To

date, 3,000 tissue-cultured Douglas firs have actually been planted in natural soil conditions.

(See figure 29.)

13D. J. Durzan, “Progress and Promise in Forest Genetics,” in

Proceedings, 50th Anniversary Symposium Paper, Science and

Technology. . . The Cutting Edge (Appleton, Wis: Institute of Paper

Chemistry, 1980).

A plantlet of Ioblolly pine grown in Weyerhaeuser Co.’s

tissue culture laboratory. The next step in this procedure

is to transfer the plant let from its sterile and humid

environment to the soil
Ch. 8—The Application of Genetics to Plants . 149

Figure 29.—A Model for Genetic Engineering

of Forest Trees

virus-free stock often appear as larger flowers,

more vigorous growth, and improved foliage

quality.

a.

f.

a.

b.

c.

d.

e.

Selection of genetic material from germplasm bank

Insertion of selected genes into protoplasts

Regeneration of cells from protoplasts and

multiplication of cell clones

Mass production of embryos from cells

Encapsulation to form ‘seeds’

f. Field germination of ‘seeds’

g. Forests of new trees

SOURCE: Office of Technology Assessment.

Creation and Maintenance of DiseaseFree Plants.—Cultivars maintained through

standard asexual propagation over long periods

often pick up viruses or other harmful path- ogens, which while they might not necessarily

kill the plants, may cause less healthy growth. A

plant’s true economic potential may be reached

only if these pathogens are removed—a task

which culturing of a plant’s meristem (growing

point) and subsequent heat therapy can perform. Not all plants produced through these

methods are virus-free, so screening cells for

viruses must be done to ensure a pathogen-free

plant. In horticultural species, the advantages of

Today, virus-free fruit plants are maintained

and distributed from both private and public

repositories. Work of commercial importance

has been done with such plants as strawberries,

sweet potatoes, citrus, freesias, irises, rhubarbs,

gooseberries, lilies, hops, gladiolus, geraniums,

and chrysanthemums. 14 Over 134 virus-free

potato cultures have also been developed by tissue culture.15

Constraints on the new genetic

technologies

Although genetic information has been transferred by vectors and protoplasm fusion, no DNA

transformations of commercial value have yet

been performed. The constraints on the successful application of molecular genetic technologies are both technical and institutional.

TECHNICAL CONSTRAINTS

Molecular engineering has been impeded by a

lack of understanding about which genes would

be useful for plant breeding purposes, as well as

by insufficient knowledge about cytogenetics.

In addition, the available tools—vectors and

mutants—and methods for transforming plant

cells using purified DNA are still limited.

Cells carrying traits important to crop productivity must be identified after they have

been genetically altered. Even if selection for an

identified trait is successful, it must be demonstrated that cells with altered properties confer similar properties on tissues, organs, and,

ultimately on the whole plant, and that the

genetic change does not adversely affect yield

or other desired characteristics. Finally, only

limited success has been achieved in regenerating whole plants from individual cells. While the

list of plant species that can be regenerated

from tissue culture has increased over the last 5

years, it includes mostly vegetables, fruit and

lqh~. NI isa~~,a, K. Sak~IO, hl. ‘1’iinaka, N1. Havashi, and H. SaIII~-

jima, “Production of Physiologically Acti\w Substances by Plant

(kll Suspension (cultures, ” H. E. SIIYXX (cd.), Tissue Culture and

Plant Science (New York: Academic Press, 1974).

jshlul,ashige, op. Cit.

76-565 0 - 81 - 11
150 “ Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

nut trees, flowers, and foliage crops. Some of

the most important crops—like wheat, oats, and

barley–have yet to be regenerated. In addition,

cells that form calluses in culture cannot always

be coaxed into forming embryos, which must

precede the formation of leaves, shoots, and

roots. Technical breakthroughs have come on a

species-by-species basis; key technical discoveries are not often applicable to all plants. And

even when the new technologies succeed in

transferring genetic information, the changes

can be unstable.

The hope that protoplasm fusion would open

extensive avenues for gene transfer between

distantly related plant species has diminished

with the observation of this instability. However, if whole chromosomes or chromosome

fragments could be transferred in plants where

sexual hybridization is presently impossible, the

possibilities would be enormous.

INSTITUTIONAL CONSTRAINTS

Institutional constraints on molecular genetics include those in funding, in regulation, in

manpower, and in industry.

Federal funding for plant molecular genetics

in agriculture has come from the National

Science Foundation (NSF) and from USDA. Research support in USDA is channeled primarily

through the flexible Competitive Grants Program (fiscal year 1980 budget of $15 million) for

the support of new research directions in plant

biology. The panel on genetic mechanisms (annual budget less than $4 million) is of particular

significance for developing new genetic technologies. The panel’s charter specifically seeks proposals on novel genetic technologies. The remaining three panels concerned with plants—

nitrogen, photosynthesis, and stress—also support projects to define the molecular basis of

fundamental plant properties. The success of

the USDA Competitive Grants Program is hard

to assess after just 2 years of operation; however, its budget over the past 2 years has severely limited expansion of the program into new

areas of research.

Some private institutions16 argue that the

W. walbot, past, present and Future Trends in Cmp f3medin&

Vol. H, Working Papers, Impact o~appiied Genetics, NTIS, 1981.

Competitive Grants Program is shifting support

from ongoing USDA programs to new genetics

research programs that are not aimed at the

important problems facing agriculture today.

There is no opposition to supporting the molecular approaches as long as they do not come at

the expense of traditional breeding programs,

and as long as both molecular biologists and

classical geneticists working with major crop

plants are assured of enough support to foster

research groups of sufficient size.

At present, funds from nine programs at the

NSF—primarily in the Directorate for Biologic],

Behavioral, and Social Sciences—support plant

research. The total support for the plant sciences may be as high as $25 million, of which

only about $1 million is designated specifically

for molecular genetics.

The regulation of the release of genetically

altered plants into the environment has not had

much effect to date. As of November 1980, only

one application [which requested exception

from the NIH Guidelines (see ch. 11) to release

rDNA-treated corn into the environment] has

been filed with the Office of Recombinant DNA

Activities (ORDA). Whether regulation will produce major obstacles is difficult to predict at

present. It is also unclear whether restrictions

will be placed on other genetic activities, such

as protoplasm fusion. Currently, at least one

other nation (New Zealand) includes such restrictions in its guidelines. It is not clear how

much the uncertainty of possible ecological disruption and the attendent liability concerns

from intentional release of genetically engineered plants has prevented the industrial sector from moving toward commercial application

of the new technology.

Only a few universities have expertise in both

plant and molecular biology. In addition, only a

few scientists work with modern molecular

techniques related to whole plant problems. As

a result, a business firm could easily develop a

capability exceeding that at any individual U.S

university. However, building industrial labora

tories and hiring from the universities could

easily deplete the expertise at the university

level. With the recent investment activity in

bioengineering firms, this trend has already
Ch. 8—The Application of Genetics to Plants ● 151

begun; in the long-run it could have serious consequences for the quality of university research.

Despite these constraints, progress in overcoming the difficulties is continuing. At the

prestigious 1980 Gordon Conference, where scientists meet to exchange ideas and recent findings, plant molecular biology was added to the

list of meetings for the first time. In addition,

four other recent meetings have concentrated

on plant molecular biology.17 Up to 50 percent

of the participants at these meetings came from

nonplant-oriented disciplines searching for future research topics. This influx of investigators

from other fields can be expected to enrich the

variety of approaches used to solve the problems of the plant breeder.

I TGenome Organization and Expression in Plants, NATO s.vmposium held in Edinburgh, Scotland, July 1979; Genetic Engineering

of Symbiotic Nitrogen Fi&ation and Conservation of Fi~ed Nitrogent

June 29-Juiy 2, 1980, Tahoe City, Ca]if; ‘“Molecular Biologists Look

al Green Plants,” Si,xfh Annual Symposium, Sepl: 29-oct. 2, lg80,

}k?idelberg, West [;ermany; and Fourth /nferna[ional Symposium

on Nitro,qm F~xation, Dec. 1-.5, 1980, Canberra, Aust ral iii.

Finally, as a general rule, tradeoffs arise in

the use of the new technologies that may interfere with their application. It is impossible to get

something for nothing from nature—e.g., in nitrogen fixation the symbiotic relationship between plant and micro-organism requires energy from the plant; screening for plants that can

produce and transfer the end products of photosynthesis to the nodules in the root more efficiently may reduce inorganic nitrogen requirements but may also reduce the overall yield.

This was the case for the high lysine varieties of

corn. (See Tech. Note 10, p. 163.) Farmers in the

United States tended to avoid them because improving the protein quality reduced the yield,

an unacceptable tradeoff at the market price.

Thus, unless the genetic innovation fits the requirements of the total agricultural industry,

potentials for crop improvement may not be

realized.

Impacts on generating new varieties

Progress in the manipulation of gene expression in eukaryotic (nucleus-containing) cells,

which include the cells of higher plants, has

been enormous. Most of the new methodologies

have been derived from fruit flies and mammalian tissue culture lines; but many should be

directly applicable to studies with plant genes.

There has been great progress in isolating specific RNA from plants, in cloning plant DNA, and

in understanding more about the organization

of plant genomes. Techniques are available for

manipulating organs, tissues, cells, or protoplasts in culture; for selecting markers; for

regenerating plants; and for testing the genetic

basis of novel traits. So far however, these

techniques are routine only in a few species.

Perfecting procedures for regenerating single

cells into whole plants is a prerequisite for the

success of many of the novel genetic technologies. In addition, work is progressing on

viruses, the Ti plasmid of Agrobacterium, and

engineered cloning vehicles for introducing

DNA into plants in a directed fashion. There

have been few demonstrations in which the inheritance of a new trait was maintained over

several sexual generations in the whole plant.

Because new varieties have to be tested

under different environmental conditions once

the problems of plant regeneration are overcome, it is difficult to assess the specific impacts

of the new technologies.—E. g., it is impossible to

determine at this time whether technical and

biological barriers will ever be overcome for

regenerating wheat from protoplasts. Nevertheless, the impact of genetics on the structure

of American agriculture can be discussed with

some degree of confidence.

Genetic engineering can affect not only what

crops can be grown, but where and how those

crops are cultivated. Although it is a variable in

production, it usually acts in conjunction with

other biological and mechanical innovations,

whose deployment is governed by social, economic, and political factors.
152 ● Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

Examples of new genetic approaches

The ways in which the new genetic approaches could aid modern agriculture are

described in the following two examples:

SELECTION OF PLANTS FOR

METABOLIC EFFICIENCY

Because terrestrial plants are immobile, they live and die

according to the dictates of the soil and weather conditions

in which they are planted; any environmental stress can

greatly reduce their yield. The major soil stresses faced by

plants include insufficient soil nutrients and water or toxic

excesses of minerals and salts. The total land area with

these conditions approaches 4 billion hectares (ha), or

about 30 percent of the land area of the Earth.

Traditionally, through the use of fertilizers, lime,

drainage, or freshwater irrigation, environments have

been manipulated to suit the plant. Modern genetic technologies might make it easier to modify the plant to suit the

environment.

Many micro-organisms and some higher plants can tolerate salt levels equal to or greater than those of sea water.

While salt tolerance has been achieved in some varieties of

plants, the classical breeding process is arduous and limited. If the genes can be identified, the possibility of actually transferring those for salt tolerance into plants makes

the adaptation of plants to high salt, semiarid regions with

high mineral toxicities or deficiencies a more feasible prospect. In the future, selecting among tissue cultures for

metabolic efficiency could become important. Tissue

culture systems could be used to select cell lines for

resistance to salts and for responsiveness to low-nutrient

levels or less fertilizer. However, too little is known about

the biochemistry and physiology of plants to allow a more

directed approach at this time. Chances for success would

be increased with a better understanding of plant cell

biology.

Such techniques could be applied to agricultural programs in less developed countries, where, commonly, supplies of fertilizers and lime are scarce, the potential for irrigation is small, and adequate support for technological

innovation is limited. [n addition, the United States itself

contains marginal land that could be exploited for forest

products and biomass. The semiarid lands of the Southwest, impoverished land in the Lake States, and reclaimed

mining lands could become cost-effective areas for production.

NITROGEN FIXATION

It has been known since the early 1800’s that biological

fixation of nitrogen is important to soil fertility. In fixation,

micro-organisms, such as the bacterium Rhizobium,

transform atmospheric nitrogen into a form that plants

can use. In some cases—e.g., with legumes this process occurs through a symbiotic relationship between the microorganism and the plant in specialized nodules on the plant

roots. Unfortunately, the major cereal crops such as

wheat, corn, rice, and forage grasses do not have the

capacity to fix atmospheric nitrogen, thus are largely

dependent on chemically produced nitrogen fertilizers.

Because of these crops, it has been estimated that the

world demand for nitrogen fertilizers will grow from 51.4

million metric tonnes (1979 estimate) to 144 million to 180

million tonnes by the year 2000.1

8 Therefore, geneticists

are looking into the possibility that the genes for nitrogen

fixation present in certain bacteria (called “nif genes”) can

be transferred to the major crops.

Laboratory investigation has focused on the molecular

biology of nitrogen fixation in the free living bacterium,

Klebsiella pneumoniae. A cluster of 15 nif genes has been

successfully cloned onto bacterial plasmids using rDNA

technology. These clones are being used to study the

molecular regulation of nif gene expression and the

physical organization of the nif genes on the Klebsiella

chromosome. In addition they have aided the search for

nitrogen fixation genes in other bacteria.

It is thought that a self-sufficient package of nitrogenfixing genes evolved during the course of plant adaptation,

and that this unit has been transferred in a functional

form to a variety of different bacterial species, including

Klebsiella and Rhizobium. If the right DNA vector can be

found, the nif genes might be transferred from bacteria to

plants. The chloroplasts, the cauliflower mosaic virus, and

the Agrobacterium Ti-plasmid are being investigated as

possible vectors.

The way that Agrobacteria, in particular, infect cells is

similar to the way Rhizobia infect plants and form

nitrogen-fixing nodules. In both cases, the physical attachment between bacterium and plant tissue is necessary for

successful infection. In the case of Agrobacteria, tumors

form when a segment of the Ti-plasmid is inserted into the

nuclear genome of the plant cell. Scientists do not yet

know exactly how a segment of the rhizobial genome is

transferred into the root tissue to induce the formation of

nodules; nevertheless, it is hoped that Agrobacteria will act

as vectors for the introduction and expression of foreign

genes into plant cells, just as Rhizobia do naturally.

Other researchers have been investigating the requirements for getting nif genes to express themselves in

plants. Nif genes from Klebsiella have already been

transferred into common yeast, an organism that can be

grown in environments without oxygen. Unfortunately,

the presence of oxygen destroys a major enzyme for

nitrogen fixation and severely limits the potential applications in higher plants. Nevertheless, it is hoped that nif

gene expression in yeast will be applicable to higher plants.

An approach that does not involve genetic engineering,

uses improved Rhizobia strains that are symbiotic with

soybeans. Through selection, Rhizobia mutants are being

found that out-perform the original wild strains. Further

‘“F. Ausubel, “Biological Nitrogen E’ixation, ” Supporting Papers:

World Food and Nurrition Study (Washington, D. C.: National Academy of Sciences, 1977).
Ch. 8—The Application of Genetics to Plants . 153

testing is needed to determine whether the improvement

can be maintained in field trials, where the improved

strains must compete against wild-type Rhizobia already

present in the soil.

Another way to improve nitrogen fixation is to select

plants that have more efficient symbiotic relationships

with nitrogen-fixing organisms. Since the biological process requires a large amount of energy from the plant, it

may be possible to select for plants that are more efficient

in producing, and then to transfer the end products of

photosynthesis to the nodules in the roots. Also existing

nitrogen-fixing bacterial strains that can interact with crop

plants which do not ordinarily fix nitrogen could be

searched for or developed.

Reducing the amount of chemically fixed nitrogen

fertilizer–and the cost of the natural gas previously used

in the chemical process—would be the largest benefit of

successfully fixing nitrogen in crops. Environmental benefits, from the smaller amount of fertilizer runoff into

water systems, would accrue as well. But is it difficult to

predict when these will become reality. Experts in the field

disagree: some feel the breakthrough is imminent; others

feel that it might take several decades to achieve.

The refinements in breeding methods provided by the new technologies may allow major

crops to be bred more and more for specialized

uses—as feed for specific animals, perhaps, or

to conform to special processing requirements.

In addition, since the populations in less developed countries suffer more often from major

nutritional deficiencies than those in industrialized countries, a specific export market of cereal grains for human consumption, like wheat

with higher protein levels, may be developed.

But genetic methods are only the tools and

catalysts for the changes in how society produces its food; financial pressures and Federal

regulation will continue to direct their course.

E.g., the automation of tissue culture systems

will decrease the labor needed to direct plant

propagation and drastically reduce the cost per

plantlet to a level competitive with seed prices

for many crops. While such breakthroughs may

increase the commercial applications of many

technologies, the effects of a displaced labor

force and cheaper and more efficient plants are

hard to predict.

Although it is difficult to make economic projections, there are several areas where genetic

technologies will clearly have an impact if the

predicted breakthroughs occur:

●

●

●

●

For

Batch culture of plant cells in automated

systems will be enhanced by the ability to

engineer and select strains that produce

larger quantities of plant substances, such

as pharmaceutical drugs.

The technologies will allow development of

elite tree lines that will greatly increase

yield, both through breeding programs

similar to those used for agricultural crops

and by overcoming breeding barriers and

lengthy breeding cycles. Refined methods

of selection and hybridization will increase

the potential of short-rotation forestry,

which can provide cellulosic substrates for

such products as ethanol or methanol.

The biological efficiency of many economically important crops will increase. Advances will depend on the ability of the

techniques to select for whole plant characteristics, such as photosynthetic soil and

nutrient efficiency.19

Besides narrowing breeding goals, the

techniques will increase the potential for

faster improvement of underexploited

plants with promising economic value.

such advances to occur, genetic factors

must be selected from superior germplasm, the

genetic contributions must be integrated into

improved cultural practices, and the improved

varieties must be efficiently propagated for

distribution.

For the soybean and tomato crops, the research area for improved biological efficiency received the highest allotment of

funds in fiscal year 1978. Total funding was $12.9 million for soybeans and $2.1 million for tomatoes. The second largest category

to be funded was control of diseases and nematodes of soybeans at

$5.1 million and for tomato at $1.6 million.
154 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Genetic variability, crop vulnerability, and

storage of germplasm

Successful plant breeding is based on the

amount of genetic diversity available for the insertion of new genes into plants. Hence, it is

essential to have an adequate scientific understanding of how much genetic erosion has taken

place and how much germplasm is needed. Neither of these questions can be satisfactorily answered today.

The amount of genetic erosion that

has taken place

Most genetic diversity is being lost because of

the displacement of vegetation in areas outside

the United States. The demand for increased

agricultural production is a principal pressure

causing deforestation of tropical latitudes (see

Tech. Note 11, p. 163), zones that contain extensive genetic diversity for both plants and

animals.

It has been estimated that several hundred

plant species become extinct every year and

that thousands of indigenous crop varieties

(wild types) have already been lost. However, it

is difficult to measure this loss, not only because

resources are on foreign soil but because erosion must be examined on a species-by-species

basis. In theory, an adequate evaluation would

require knowledge of both the quantity of diversity within a species and the breadth of that

diversity; this process has in practice, just begun. What is known is that the lost material cannot be replaced.

The amount of germplasm needed

Germplasm is needed as a resource for improving characteristics of plants and as a means

for guaranteeing supplies of known plant

derivatives and potential new ones. Even if

plant breeders adequately understand the

amount of germplasm presently needed, it is difficult to predict future needs. Because pests and

pathogens are constantly mutating, there is

always the possibility that some resistance will

be broken down. Even though genetic diversity

can reduce the severity of economic loss, an epidemic might require the introduction of a new

resistant variety. In addition, other pressures

will determine which crops will be grown for

food, fiber, fuel, and pharmaceuticals, and how

they will be cultivated; genetic diversity will be

fundamental to these innovations.

Even if genetic needs can be adequately identified, there is disagreement about how much

germplasm to collect. In the past, its collection

has been guided by differences in morphology

(form and structure), which have not often been

directly correlated to breeding objectives. Furthermore, the extent to which the new genetic

technologies will affect genetic variability, vulnerability, or the storage of germplasm, has not

been determined. (See app. II-A.)

In addition to its uses in plant improvement,

germplasm can provide both old and new products. Recent interest in growing guayule as a

source of hydrocarbons (for rubber, energy

materials, etc.) has focused attention on plants

that may possibly be underutilized. It has been

found that past collections of guayule germplasm have not been adequately maintained,

making current genetic improvements more difficult. In addition, half of the world’s medicinal

compounds are obtained from plants; maintaining as many varieties as possible would ensure

the availability of compounds known to be useful, as well as new, and as yet undiscovered

compounds—e.g., the quinine drugs used in the

treatment of malaria were originally obtained

from the Cinchona plant. A USDA collection of

superior germplasm established in 1940 in

Guatemala was not maintained. As a conse-
Ch. 8—The Application of Genetics to Plants ● 155

quence, difficulties arose during the Vietnam

War when the new antimalarial drugs became

less effective on resistant strains of the parasite

and natural quinines were once again used.

An important distinction exists between preserving genetic resources in situ and preserving

germplasm stored in repositories. Although

genetic loss can occur at each location, evolution will continue only in natural ecosystems.

With better storage techniques, seed loss and

genetic “drift” can be kept to a minimum. Nevertheless, species extinction in situ will continue.

The National Germplasm System

USDA has been responsible for collecting and

cataloging seed (mostly from agriculturally important plants) since 1898. Yet it is important to

realize that other Federal agencies also have

responsibilities for gene resource management.

(See table 27.) Over the past century, over

440,000 plant introductions from more than 150

expeditions to centers of crop diversity have

been cataloged.

The expeditions were needed because the

United States is gene poor. The economically imTable 27.—Gene Resource Responsibilities of Federal Agencies

Type of ecosystems

under Federal

Agency ownership/control Responsibilities

U.S. Department of Agriculture

Animal & Plant Health Inspection

Service . . . . . . . . . . . . . . . . . . . . . . . . —

Forest Service . . . . . . . . . . . . . . . . . . . . Forestlands and

rangelands (U.S.

National Forest)

Science & Education Administration . —

Soil Conservation Service. . . . . . . . . . . —

Controls insect and disease problems of commercially

valuable animals and plants.

Manages forest land and rangeland living resources for

production.

Develops animal breeds, crop varieties, and microbial strains.

Manages a system for conserving crop gene resources.

Develops plant varieties suitable for reducing soil erosion and

other problems.

Department of Commerce

National Oceanic & Atmospheric

Administration. . . . . . . . . . . . . . . . . . . Oceans—between 3 Manages marine fisheries.

Department of Energy. . . . . . . . . . . . . . . .

Department of Health& Human Services

National Institutes of Health . . . . . . . .

Department of the Interior

Bureau of Land Management. . . . . . . .

Fish & Wildlife Service . . . . . . . . . . . . .

National Park Service . . . . . . . . . . . . . .

Department of State . . . . . . . . . . . . . . . . .

Agency for International Development. .

Environmental Protection Agency. . . . . .

National Science Foundation. . . . . . . . . .

and 200 miles off

the U.S. coasts

— Develops new energy sources from biomass.

— Utilizes animals, plants, and micro-organisms in medical

research.

Forest lands, Manages forest, range, and desert living resources for

rangelands, and production.

deserts

Broad range of Manages game animals, including fish, birds, and mammals.

habitats, including

oceans up to 3 miles

off U.S. coasts

Forest lands, Conserves forestland, rangeland, and desert-living resources.

rangelands, and

deserts (U.S.

National Parks)

— Concerned with international relations regarding gene

resources.

— Assists in the development of industries in other countries

including their agriculture, forestry, and fisheries.

— Regulates and monitors pollution.

— Provides funding for genetic stock collections and for research

related to gene resource conservation.

SOURCE: David Kapton, National Assoclatlon for Gene Resource Conservation.
156 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

portant food plants indigenous to the continental United States are limited to the sunflower,

cranberry, blueberry, strawberry, and pecan.

The centers of genetic diversity, found mostly in

tropical latitudes around the world, are believed to be the areas where progenitors of major crop plants originated. Today, they contain

genetic diversity that can be used for plant improvement.

It is difficult to estimate the financial return

from the germplasm that has been collected,

but its impact on the breeding system has been

substantial. A wild melon collected in India, for

instance, was the source of resistance to powdery mildew and prevented the destruction of

California melons. A seemingly useless wheat

strain from Turkey—thin-stalked, highly susceptible to red rust, and with poor milling properties—was the source of genetic resistance to

stripe rust when it became a problem in the

Pacific Northwest. Similarly, a Peruvian species

contributed “ripe rot” resistance to American

pepper plants, while a Korean cucumber strain

provided high-yield production of hybrid cucumber seed for U.S. farmers. And a gene for

resistance to Northern corn blight transferred

to Corn Belt hybrids has resulted in an estimated savings of 30 to 50 bushels (bu) per acre,

with a seasonal value in excess of $200 million.20

(See table 28.)

The effort to store and evaluate this collected

germplasm was promoted by the Agricultural

Marketing Act of 1946, which authorized regional and interregional plant introduction stations (National Seed Storage Centers) run cooperatively by both Federal and State Governments. The federally controlled National Seed

Storage Laboratory in Fort Collins, Colo., was

established in 1958 to provide permanent storage for seed. In the 1970’s, it was recognized

that the system should include clonal material

for vegetatively propagated crops, which cannot be stored as seed. Although their storage requires more space than comparable seed storZOu. s, Department of Agriculture, Agricultural Research SerVice, Introduction, Class[~ication, Maintenance, Evaluation, and Documentation of Plant Germplasm, (ARS) National Research Program

No. 20160 (Washington, D. C., U.S. Government Printing Office,

1976).

Table 28.-Estimated Economic Rates of Return

From Germplasm Accessions

1. A plant introduction of wheat from Turkey was found to

2.

3.

4.

—

have resistance to all known races of common and

dwarf bunts, resistance to stripe rust and flag smut,

plus field resistance to powdery and snow mold. It has

contributed to many commercial varieties, with

estimated annual benefits of $50 million.

The highly successful variety of short-strawed wheat,

‘Gaines’ has in its lineage three plant introductions that

contributed to the genes for the short stature and for

resistance to several diseases. During the 3 years,

1964-66, about 60 percent of the wheat grown in the

State of Washington was with the variety ‘Gaines’. increased production with this variety averaged slightly

over 13 million bu or $17.5 million per year in the 3-year

period.

Two soybean introductions from Nanking and China

were used for large-scale production, because they are

well-adapted to a wide range of soil conditions. All major soybean varieties now grown in t e Southern United

States contain genes from one or both of these introductions. Farm gate value of soybean crop in the

South exceeded $2 billion in 1974.

Two varieties of white, seedless grapes resulted from

crosses of two plant introductions. These varieties

ripen 2 weeks ahead of ‘Thompson Seedless’. Benefits

to the California grape industry estimated to be more

than $5 million annually.

SOURCE: U.S. Department of Aorlculture. Agricultural Research Service. /ntroduct;on, Classitica~ion, Maintenance, Eveiuation, and Documentation of Piant Gerrrrpiasrrr, (ARS) National Research Program No.

20160 (Washington, D. C., U.S. Government Printing Of fice,1976).

age, 12 new repositories for fruit and nut crops

as well as for other important crops, from hops

to mint, were proposed by the National Germplasm Committee as additions to the National

Germplasm System (see Tech. Note 12, p. 163).

(The development of tissue culture storage

methods may reduce storage costs for these

proposed repositories.)

The National Germplasm System is a vital link

in ensuring that germplasm now existing will

still be available in the future. However, the

present system was challenged after the Southern corn blight epidemic of 1970. Many scientists questioned whether it was large enough

and broad enough in its present form to provide

the genetic resources that might be needed.

The devastating effects of the corn blight of

1970 actually led to the coining of the term crop

vulnerability. During the epidemic, as much as

15 percent of the entire yield was lost. Some

fields lost their whole crop, and entire sections

of some Southern States lost 50 percent of their
Ch. 8—The Application of Genetics to Plants . 157

corn. Epidemics like this one are, of course, not

new. In the 19th century, the phylloxera disease

of grapes almost destroyed the wine industry of

France, coffee rust disrupted the economy of

Ceylon, and the potato famine triggered extensive local starvation in Ireland and mass emigration to North America. In 1916, the red rust destroyed 2 million bu of wheat in the United

States and an additional million in Canada. Further epidemics of wheat rust occurred in 1935

and 1953. The corn blight epidemic in the

United States stimulated a study that led to the

publication of a report on the “Genetic Vulnerability of Major Crops’’. 21 It contained two central findings: that vulnerability stems from genetic uniformity, and that some American crops

are, on this basis, highly vulnerable. (See table

29.)

However, genetic variability, is only a hedge

against vulnerability. It does not guarantee that

an epidemic will be avoided. In addition, pathogens from abroad can become serious problems when they are introduced into new environments. As clearly stated in the study, a triangular relationship exists between host, pathogen, and environment, and the coincidence of

their interaction dictates the severity of disease.

z INati~nal Academy of Sciences, Genetic Vulnerability of M@r

Crops, Washington, D. C., 1972.

The basis for genetic uniformity

Crop uniformity results most often from societal decisions on how to produce food. The

structure of agriculture is extremely sensitive to

changes in the market. Some of the basic factors

influencing uniformity are:

the consumer’s demand for high-quality

produce;

the food processing industry’s demand for

harvest uniformity;

the farmer’s demand for the “best” variety

that offers high yields and meets the needs

of a mechanized farm system; and

the increased world demand for food,

which is related to both economic and population growth.

New varieties of crops are bred all the time,

but several can dominate agricultural production—e.g., Norman Borlaug and his colleagues in

Mexico pioneered the “green revolution” by

developing high-yielding varieties (HYV) of

wheat that required less daylight to mature and

possessed stiffer straw and shorter stems. Since

the new varieties (see Tech. Note 13, p. 163)

gave excellent yields in response to applications

of fertilizer, pesticides, and irrigation, the innovation was subsequently introduced into

countries like India and Pakistan. When a single

Table 29.—Acreage and Farm Value of Major U.S. Crops and Extent to Which

Small Numbers of Varieties Dominate Crop Average (1969 figures)

Value

Acreage (millions of Total Major Acreage

Crop (millions) dollars) varieties varieties (percent)

Bean, dry . . . . . . . . . . . . . . . . . . 1.4 143 25 2 60

Bean, snap. . . . . . . . . . . . . . . . . 0.3 99 70 3 76

Cotton. . . . . . . . . . . . . . . . . . . . . 11.2

Coma. . . . . . . . . . . . . . . . . . . . . .

1,200 50 3 53

66.3 5,200 197b 6

C

71

Millet. . . . . . . . . . . . . . . . . . . . . . 2.0 ? 3 100

Peanut . . . . . . . . . . . . . . . . . . . . 1.4 312 15 9 95

Peas . . . . . . . . . . . . . . . . . . . . . . 0.4 80 50 2 96

Potato. . . . . . . . . . . . . . . . . . . . . 1.4 616 82 4 72

Rice. . . . . . . . . . . . . . . . . . . . . . . 1.8 449 14 4 65

Sorghum. . . . . . . . . . . . . . . . . . . 16.8 795 ? ? ?

Soybean . . . . . . . . . . . . . . . . . . . 42.4 2,500 62 6 58

Sugar beet . . . . . . . . . . . . . . . . . 367 16 2 42

Sweet potato . . . . . . . . . . . . . . . 0.13 63 48 69

Wheat. . . . . . . . . . . . . . . . . . . . . 44.3 1,800 269 9 50

acorn k-iclldeg seeds, forage, and sila9e.

bRelea9ed public inbreds only.

cThere were six major public lines used in breeding the major varieties of corn, so the aCtIJal number of ‘farietieS is higher.

SOURCE: National Academy of Sciences, Genetic Vu/nerabi/ity of Major Crops, Washington, D. C., 1972.
I&l . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

variety dominates the planting of a crop, there

is some loss of genetic variability, the resulting

uniformity causes crop vulnerability—and the

displacement of indigenous varieties—a real

problem.

The rate of adoption of HYVs levels off below

100 percent in most countries, mainly because

of the numerous factors affecting supply and

demand: 22

● supply factors:

—the present HYVs are not suitable for all

soil and climatic conditions;

—they require seeds and inputs (such as

fertilizers, water, and pesticides) that are

either unavailable or not fully utilized by

every farmer; and

—in some regions, a strong demand still exists for the longer straw of traditional

varieties.

● demand factors:

—consumers may not prefer the HYVs over

traditional food varieties;

—Government price policies may not encourage the production of HYVs.

For these and other reasons, countries already

using a great deal of HYVs will continue to adopt

them more slowly.

Six factors affecting adequate

management of genetic resources

1. Estimating the potential value of genetic resources is dfficult.

Of the world’s estimated 300,000 species of

higher plants, only about 1 percent have been

screened for their use in meeting the diverse

demands for food, animal feed, fiber, and pharmaceuticals. 23 Genetic resources not yet collected or evaluated are valuable until proven

otherwise, and the efforts to conserve, collect,

and evaluate plant resources should reflect this

assumption. This point of view was strongly reflected in a 1978 recommendation by the National Plant Genetic Resources Board. It’s recommendation was that four major areas of genetic

storage—collection, maintenance, evaluation,

and distribution—be viewed as a “continuum

that sets up a gene flow from source to end

use".24

Z. The management of genetic resources is complex and costly.

The question of how much germplasm to collect is difficult and strongly influenced by cost.

Thus far, only a fraction of the available diversity has been collected. A better scientific understanding of the genetic makeup and previous

breeding history of major crops will help determine just how much germplasm should be collected. Efforts to give priorities for co11ection25

have been hindered by the scientific gaps in

knowledge about what is presently stored

worldwide. And while attempts have been

made to estimate the economic return from introduction of specific plants (see table 28), the

degree to which agricultural production and

stability are dependent on genetic variability

has not been adequately analyzed.

Evaluation of genetic characteristics must be

conducted at different ecological sites by multidisciplinary teams. The data obtained will only

be useful if adequately assessed and made available to the breeding community (see Tech. Note

14, p. 163).

Germplasm must be adequately maintained to

assure viability, “working stocks” must be made

available to the breeding community. The primary objective of storing germplasm is to make

the genetic information available to breeders

and researchers.

3. How much plant diversity can be lost

disrupting the ecological balances of

and agricultural systems is not known.

without

natural

=D. G. Dalrymp]e, Oeve/opment and Spread o~Wgh-k’ielding VWieties of Wheat and t?ice in the Less Developed Nations, 6th ed.

(Washington D. C.: U.S. Department of Agriculture, Office of International Cooperation and Development in cooperation with U.S.

Agency for International Development, 1978).

2SN. Myers, “[conserving Our Global stock, ” Environment

21(9):25, 1979.

z4RepOr[ to the Secretarv of Agriculture, bY the Assistant seeretary for Conservation, R~search, and Education based on the deliberations and recommendations, Nationai Plant Genetic Resources Board, July 1978.

Zssecretaria[, International Board for Plant Genetic Resources,

Annual Report 1978, Rome; Consultative Group on International

Agricultural Research, 1979.
Ch. 8—The Application of Genetics to Plants ● 159

The arguments parallel those previously discussed in Congress for protection of endangered species (see Tech. Note 15, p. 163). The

last decade has shown that modes of production

and development can severely affect the ecological balance of complex ecosystems. What is not

known is how much species disruption can take

place before the quality of life is also affected.

4. The extent to which the new genetic technologies will affect genetic variability, germplasm

storage methodologies, and crop vulnerability

has not been determined.

The new genetic technologies could either increase or decrease crop vulnerability. In theory,

they could be useful in developing early warning systems for vulnerability by screening for

inherent weaknesses in major crop resistance.

However, the relationship between the genetic

characteristics of plant varieties and their pests

and pathogens is not understood (see Tech. Note

16, p. 164).

The new technologies may also enhance the

prospects of using variability, creating new

sources of genetic diversity and storing genetic

material by:

●

●

●

●

increasing variability during cell regeneration,

incorporating new combinations of genetic

information during cell fusion,

changing the ploidy level of plants, and

introducing foreign (nonplant) material

and distantly related plant material by

means of rDNA.

With the potential benefits, however, come

risks. Because genetic changes during the development of new varieties are often cumulative,

and because superior varieties are often used

extensively, the new technologies could increase both the degree of genetic uniformity

and the rate at which improved varieties displace indigenous crop types. Furthermore, it

has not been determined how overcoming natural breeding barriers by cell fusion or rDNA will

affect a crop’s susceptibility to pests and diseases.

5. Because pests and pathogens are constantly

mutating plant resistance can be broken down,

requiring the introduction of new varieties.

Historically, success and failure in breeding

programs are linked to pests and pathogens

overcoming resistance. Hence, plant breeders

try to keep one step ahead of mutations or

changes in pest and pathogen populations; a

plant variety usually lasts only 5 to 15 years on

the market. There is some evidence that pathogens are becoming more virulent and aggressive—which could increase the rate of infection,

enhancing the potential for an epidemic (see

Tech. Note 17, p. 164).

6. Other economic and social pressures affect the

use of genetic resources.

The Plant Variety Protection Act has been

criticized for being a primary cause of planting

uniform varieties, loss of germplasm, and conglomerate acquisition of seed companies. In its

opponents’ view, such ownership rights provide

a strong incentive for seed companies to encourage farmers to buy “superior” varieties that

can be protected, instead of indigenous varieties

that cannot. They also make plant breeding so

lucrative that the ownership of seed companies,

is being concentrated in multinational corporations—e.g., opponents claim that 79 percent of

the U.S. patents on beans have been issued to

four companies and that almost 50 once-independent seed companies have been acquired by

The Upjohn Co., ITT, and others.26 One concern

raised about such ownership is that some of

these companies also make fertilizer and

pesticides and have no incentive to breed for

pest resistance or nitrogen-fixation. For the

above reasons, one public interest group has

concluded :27

[t]hanks to the patent laws, the bulk of the

world’s food supply is now owned and developed by a handful of corporations which alone,

without any public input, determine which

strains are used and how.

Numerous arguments have been advanced

against the above position. Planting of a single

variety, for instance, is claimed to be a function

of the normal desires of farmers to purchase

the best available seed, especially in the com2’P. R. Mooney, Seed of the Earfh (London: International

Coalition tor De\7elopment Action, 1979).

2T~1.1ef for PeoP]eS’ ~usilless (hmm ission as Amicus Curiae ,

Diamond v. Chalmabarr-v, 100 S. Ct. 2204 (1980), p. 9.
160 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

petitive environment in which they operate.

Moreover, hybrid varieties (such as corn), are

not covered by the plant protection laws; yet

they comprise about 90 percent of the seed

trade.

As for the loss of varieties by vegetation

displacement, statutory protection has been too

recent to counter a phenomenon that has occurred over a 30- to 40-year period, and available evidence indicates that some crops are actually becoming more diverse. Since most major

food crops are sexually produced, they have

only been subject to protection since 1970 when

the Plant Variety Protection Act was passed; the

first certificates under that Act were not even

issued until 1972. Moreover, at least in the case

of wheat, as many new varieties were developed in the 7 years after the passage of the Plant

Variety Protection Act as in the previous 17.28

It is clear that large corporations have been

acquiring seed companies. However, the con20H. Rept. No. 96.1115, 96th Cong., 2d S6?SS., p. 5 (June 20J 1980).

nection between this trend and the plant variety

protection laws is disputed. one explanation is

that the takeovers are part of the general takeover movement that has involved all parts of the

economy during the past decade. Since the passage of the 1970 Act, the number of seed companies, especially soybean, wheat, and cereal

grains, has increased.29 While there were six

companies working with soybean breeding

prior to 1970, there are 25 at this time.30

Thus, to date, although no conclusive connection has been demonstrated between the two

plant protection laws and the loss of genetic

diversity, the use of uniform varieties, or the

claims of increasing concentration in the plant

breeding industry; the question is stilI controversial and these complex problems are still

unresolved.

ZgHearin@ on H.R. 2844, supra note 35 (Statement of’ Harold

Loden, Executive Director of the American Seed Trade Association).

JoBrief for pharmace~tica] Manufacturers’ Association as

Amicus Curiae, DiarnorId v. Cha)crabarty, 100 S. Ct. 2204 (1980), p.

26.

Summary

The science and structure of agriculture are

not static. The technical and industrial revolutions and the population explosion have all contributed to agricultural trends that influence

the impacts of the new technologies. Several

factors affect U.S. agriculture in particular:

To some degree, the United States depends

on germplasm from sources abroad, which

are, for the most part, located in less developed countries; furthermore, the amount

of germplasm from these areas that should

be collected has not been determined.

Genetic diversity in areas abroad is being

lost. The pressures of urbanization, industrial development, and the demands for

more efficient, more intensive agricultural

production are forcing the disappearance

of biological natural resources in which the

supply of germplasm is maintained.

● This lost genetic diversity is irreplaceable.

● The world’s major food crops are becoming

more vulnerable as a result of genetic uniformity.

The solutions—examining the risks and evaluating the tradeoffs—are not limited to securing

and storing varieties of seed in manmade

repositories; genetic evolution—one of the keys

to genetic diversity and a continuous supply of

new germplasm—cannot take place on storage

shelves. Until specific gaps in man’s understanding of plant genetics are filled, and until the

breeding community is able to identify, collect,

and evaluate sources of genetic diversity, it is

essential that natural resources providing germplasm be preserved.
Ch. 8—The Application of Genetics to Plants ● 161

Issues and Options—Plants

ISSUE: Should an assessment be conducted to determine how much

plant germplasm needs to be

maintained?

An understanding of how much germplasm

should be protected and maintained would

make the management of genetic resources

simpler. But no complete answers exist; nobody

knows how much diversity is being lost by

vegetation displacement in areas mostly outside

the United States.

OPTIONS:

A. Congress could commission a study on how

much genetic variability is needed or desirable

to meet present and future needs.

A comprehensive evaluation of the National

Germplasm System’s needs in collecting, evaluating, maintaining, and distributing genetic

resources for plant breeding and research could

serve as a baseline for further assessment. This

evaluation would require extensive cooperation

among the Federal, State, and private components linked to the National Germplasm System.

B. Congress could commission a study on the

need for international cooperation to manage

and preserve genetic resources both in natural

ecosystems and in repositories.

This investigation could include an evaluation

of the rate at which genetic diversity is being

lost from natural and agricultural systems, and

an estimate of the effects this loss will have. Until such information is at hand, Congress could:

● Instruct the Department of State to have its

delegations to the United Nations Educational, Scientific, and Cultural Organization

(UNESCO) and United Nations Environmental Program (UNEP) encourage efforts to establish biosphere reserves and other protected natural areas in less developed countries, especially those within the tropical

latitudes. These reserves would serve as a

source for continued natural mutation and

variation.

●

●

●

Instruct the Agency for International Development (AID) to place high priority on,

and accelerate its activities in, assisting less

developed countries to establish biosphere

reserves and other protected natural areas,

providing for their protection, and support

associate research and training.

Instruct the International Bank for Reconstruction and Development (World Bank) to

give high priority to providing loans to

those less developed countries that wish to

establish biosphere reserves and other protected natural areas as well as to promote

activities related to biosphere reserve preservation, and the research and management of these areas and resources.

Make a one-time special contribution to

UNESCO to accelerate the establishment of

biosphere reserves.

Such measures for in situ preservation and

management are necessary for long-term maintenance of genetic diversity. Future needs are

difficult to predict; and the resources, once lost

are irreplaceable.

C. Congress could commission a study on how to

develop an early warning system to recognize

potential vulnerability of crops.

A followup study to the 1972 National Academy of Science’s report on major crop vulnerability could be commissioned. Where high

genetic uniformity still exists, proposals could

be suggested to overcome it. In addition, the

avenues by which private seed companies could

be encouraged to increase the levels of genetic

diversity could be investigated. The study could

also consider to what extent the crossing of

natural breeding barriers as a consequence of

the new genetic technologies will increase the

risks of crop vulnerability.

ISSUE: What are the most appropriate

approaches for overcoming the

various technical constraint s

that limit the success of molecular genetics for plant improvement?
162 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Although genetic information has been transferred by vectors and protoplasm fusion, DNA

transformations of commercial value have not

yet been performed. Molecular engineering has

been impeded by the lack of vectors that can

transfer novel genetic material into plants, by

insufficient knowledge about which genes

would be useful for breeding purposes, and by

a lack of understanding of the incompatibility of

c.

sis on potential pharmaceuticals derived from

plants.

Establish an institute for plant molecular genetics under the Science and Education Administration at USDA that would include multidisciplinary teams to consider both basic research questions and direct applications of the

technology to commercial needs and practices.

chromosomes from diverse sources. Another The discoveries of molecular plant genetics

impediment has been the lack of researchers will be used in conjunction with traditional

from a variety of disciplines. breeding programs. Therefore, each of the

three options would require additional approOPTIONS: priations for agricultural research. Existing

funding structures could be used for all three, A. Increase the level of funding for plant molec- but institutional reorganization would be re- ular genetics through:

1. the National Science Foundation (NSF)) and quired for options B and C. The main argument

Z. the Competitive Grants Program of the U.S. for increasing USDA funding is that it is the lead

Department of Agriculture (USDA). agency for agricultural research, for increasing

NSF and NIH funding, that they currently have

B. Establish research units devoted to plant mo- the greatest expertise in molecular techniques.

lecular genetics

tional Institutes

Technical

under the auspices of the Na- Option C emphasizes the importance of the inof Health (NIH), with empha- terdisciplinary needs of this research.

notes

1. A recent example of such a mutation was the opaque-2

gene in corn, which was responsible for increasing the

corn’s content of the amino acid lysine.

Z. There is disagreement about what is meant by productivity and how it is measured. Statistical field data can

be expressed in various ways—e.g., output per manhour, crop yield per unit area, or output per unit of

total inputs used in production. A productivity measurement is a relationship among physical units of production. It differs from measurements of efficiency,

which relate to economic and social values.

3. Nevertheless, some parts of the world continue to lack

adequate supplied of food. A recent study by the Presidential Commission on World Hunger31 estimates that

“at least one out of every eight men, women, and children on earth suffers malnutrition severe enough to

shorten life, stunt physical growth, and dull mental

ability. ”

4. In theory, pure lines produce only identical gametes,

which makes them true breeders. Successive crossbreeding will result in a mixture of gametes with varying combinations of genes at a given locus on homologous chromosomes.

31 RePO1.t of the pr~sidelltia] (;ornnlission on World Hungfwt ~~v~rcomin$ World Hunger: The L’hallenge Ahead, Washington, D. C.,

March 1980.

5

6

7

Normally, chromosomes are inherited in sets. The

more frequent diploid state consists of two sets in each

plant. Because chromosome pairs are homologous

(have the same linear gene sequence), cells must maintain a degree of genetic integrity between chromosome

pairs during cell division. Therefore, increases in

ploidy involve entire sets of chromosomes—diploid (2-

set) is manipulated to triploid (3-set) or even to

tetraploid (4-set).

The estimated theoretical limit to efficiency of photosynthesis during the growth cycle is 8.7 percent. However, the record U.S. State average (116 bu/acre, Illinois, 1975) for corn, having a high photosynthetic rate

in comparison to other major crops, approaches only 1

percent efficiency. 32 Since a major limiting step in plant

productivity lies in this efficiency for the photosynthetic process, there is potential for plant breeding

strategies to improve the efficiency of photosynthesis

of many other important crops. This would have a tremendous impact on agricultural productivity.

It is difficult to separate social values from the economic structures affecting the productivity of American

agriculture. Social pressures and decisions are complex

iZoffi~.~ of ‘[”~~:hll[]]t)gy /l SSPSSlllPllt, 11.S. Congress, Energv for

Bioiogicai Proresses, Volume 11: Technical Analysis (W’ashinglon,

11.(1,: ~1.S. (;mwrnment Printing office, Ju]y 1980).
Ch. 8—The Application Of Genetics to Plants ● 163

8.

9.

10.

11

and integrated—e.g., conflicts between developing

maximum productivity and environmental concerns

are typified by the removal of effective pesticides from

the market. Applications of existing or new technologies may be screened by the public for acceptable

environmental impact. Conflict also exists between

higher productivity and higher nutritive content in

food, since selection for one often hurts the other.

A critical photosynthetic enzyme (ribulose biphosphate

carboylase) is formed from information supplied by

different genes located independently in the chloroplast (a plastid) and the nucleus of the cell. It is composed of two separate protein chains that must link

together within the chloroplast. The larger of these

chains is coded for by a gene in the chloroplast—and it

is this gene that has been recently isolated and cloned.

The smaller subunit, however, derives from the plant

nucleus itself. This cooperation between the nucleus

and the chloroplast to produce the functional expression of a gene is an interesting phenomenon. Because it

exists, the genetics of the cell could be manipulated so

that cytoplasmically introduced genes can influence

nuclear gene functions. Perhaps most importantly at

this stage, plastid genes are prime candidates to clarify

the basic molecular genetic mechanisms in higher

plants.

The advantages to using mass propagation techniques

for strawberry plants are that those produced from

tissue culture are virus-free, and a plantlet produced in

tissue culture can produce more shoots or runners for

transplanting.

The disadvantages are that during the first year the

fruit tends to be smaller and, therefore, less commercially acceptable; the plants from tissue culture may

have trouble adapting to soil conditions, which can affect their vigor, especially during the first growing season; and the price per plantlet ready for planting from

tissue culture systems may be more expensive than

commercial prices for rooted shoots or runners bought

in bulk.

Wheat protein is deficient in several amino acids, including lysine. Considerable attention has been devoted in the past 5 to 10 years to improving the nutritional properties of wheat. Thousands of lines have

been screened for high protein, with good success, and

high lysine genes with poor success. Some high protein

varieties have been developed, but adoption by the

farmer has been mediocre at best, partly because of

reduced yield levels. There are some “exceptions; —e.g.,

the Variety “Plainsman V“ has maintained both high

protein and yield levels, which indicates that there is no

consistent relationship between low protein and high

yields in some varieties.

Some 42 percent of the total land area in the tropics,

consisting of 1.9 billion hectares, contains significant

forest cover. It is difficult to measure precisely the

amount of permanent forest cover that is being lost;

however, it has been estimated that 40 percent of

“closed” forest (having a continuous closed canopy) has

already been lost, with 1 to 2 percent cleared annually.

12

13

14

15.

If the highest predicted rate of loss continues, half of

the remaining closed forest area will be lost by the year

2000. 33 The significance of this loss is expressed by

Norman Myers in his report, Conversion of Tropical

Moist Forests, prepared for the Committee on Research

Priorities in Tropical Biology of the National Academy

of Science’s National Research Council: “Extrapolation

of figures from well-known groups of organisms suggest that there are usually twice as many species in the

tropics as temperate regions. If two-thirds of the

tropical species occur in TMF (tropical moist forests), a

reasonable extrapolation from known relationships,

then the species of the TMF should amount to some 40

to so percent of the planet’s stock of species—or somewhere between 2 million and 5 million species altogether. In other words, nearly half of all species on Earth

are apparently contained in a biome that comprises

only 6 percent of the globe’s land surface. Probably no

more than 300,000 of these species—no more than 15

percent and possibly much less—have ever been given

a Latin name, and most are totally unknown. "34

In 1975, the Committee estimated that $4 million would

be necessary for capital costs of each repository, with

recurring annual expenses of $1.4 million for salaries

and operations. USDA has allocated $1.16 million for its

share of the construction costs for the first facility to

be constructed at the Oregon State University in Corvallis.

High yielding varieties (HYVs) can be defined as potentially high-yielding, usually semidwarf (shorter than

conventional), types that have been developed in national research programs worldwide. Wheat varieties

were developed by the International Maize and Wheat

Improvement Center and rice varieties by international Rice Research Institute. Many improved varieties

of major crops of conventional height are not currently

considered HYV types, but they have often been incorporated into HYV breeding. HYVs, because of biological

and management factors, rarely reach their full

harvest potential.

Although the National Germplasm System successfully

handles some 500,000 units to meet annual germplasm

requests, many accessions–like the 35,000 to 40,000

wheat accessions stored at the Plant Genetics and

Germplasm Institute at Beltsville, Md.–have yet to be

examined. Furthermore, the varieties released for sale

by the seed companies are not presently evaluated for

their comparative genetic differences.

For comparison, the National Germplasm System functions on less than $10 million annually, whereas the Endangered Species Program had a fiscal year 1980 budget of over $23 million. The funds allocated to the EnjsRep~rt t. the Prmident bv a 11 .S. Interagency Task Force 011

Tropical Forests, 7’he Worldk Tropical Forests: A Policy, S[rategv,

and Program for the C hited Slates, State Departnlent publication

No. 9117, }Vashin#on, [1.(; ., hla~~, 1980.

W,V, ~lver.s, [;onktersjon Of ‘rro~jcal Moist Forests, report for the

Conmitiee on Research Priorities in Tropical Biology of the National Research Council, National Academy of Sciences, Washington, D.(:., 1980.
164 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

dangered Species Program are used for such activities

as listing endangered species, purchasing habitats for

protection, and law enforcement.

16. The uses of pest-resistant wheat and corn cultivars on

a large scale for both diseases and insects are classic

success stories of host- plant resistance. However, recent trends in the Great Plains Wheat Belt are disturbing. The acreage of Hessian fly-resistant wheats in Kansas and Nebraska has decreased from about 66 percent

in 1973 to about 42 percent in 1977. Hessian fly infestations have increased where susceptible cultivars have

been planted. In South Dakota in 1978, in an area not

normally heavily infested, an estimated 1.25 million

acres of spring wheat were infested resulting in losses

of $25 million to $50 million. An even greater decrease

in resistant wheat acreage is expected in the next 2 to 5

years as a result of releases of cultivars that have improved agronomic traits and disease resistance but that

are susceptible to the Hessian fly. Insect resistance has

not been a significant component of commercial

breeding programs.35

~sotfi{:[l ~,f ‘l’(~chno]o~v Assessment, 11. S. (Mgl’ess, Pest Managernenf Sfra~egies in Crop Protection (vol. 1, Washington, D. C.: IJ. S.

(kn’ernnwnt Printing office, octoher 1979), p. 73.

17. Expressed in genetic terms, cases exist ‘(where the introduction of novel sources of major gene resistance

into commercial cultivars of crop plants has resulted in

an increase in their frequency of corresponding virulence genes in the pathogen’ ’.” This has been reported

in Australia with wheat stem rust, barley powdery mildew, tomato leaf mold, and lettuce downy mildew. Evidence suggests that there is considerable gene flow in

the various pathogen populations-e.g., asexual transfer can quickly alter the frequency of virulence genes.

Furthermore, pressures brought about in the evolutionary process have developed such a high degree of

complexity in both resistance and virulence mechanisms, that breeding approaches, especially those only

using single gene resistance, can be easily overcome.

36R, ~;. Shattock, B. ~. Janss~n, R, Whitbread, and D. S. Shaw,

“h Interpretation of the Frequencies of Host Specific Phenotypes

of Phytophthora infestans in North Wales, ” Ann. Appli. Biol. 86:249,

1977.
chapter 9

Advances in Reproductive

Biology and Their Effects

on Animal Improvement
Chapter 9

Page

Background . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167

The Scientific Era in Livestock Production . . . . . 167

Controlled Breeding. . . . . . . . . . . . . . . . . . . . . 168

Scientific Production . . . . . . . . . . . . . . . . . . . . 170

Resistance to Change . , , . . . . . . . . . . . . . . . . . 171

Some Future Trends . . . . . . . . . . . . . . . . . . . . 172

Technologies. . . . . . . . . . . . . . . . . . . . . . . . . . . . 173

Technologies That Are Presently Useful. . . . . . 174

Sperm Storage . . . . . . . . . . . . . . . . . . . . . . . 174

Artificial Insemination . . , . . . . . . . . . . . . . . 174

Estrus Synchronization. . . . . . . . . . . . . . . . . 176

Superovulation , . . . . . . . . . . . . . . . . . . . . . , 176

Embryo Recovery . . , . . . . . . . . . . . . . . . . . . 176

Embryo Transfer . . . . . . . . . . . . . . . . . . . . . 177

Embryo Storage . . . . . . . . . . . . . . . . . . . . . . 177

Sex Selection. . . . . . . . . . . . . . . . . . . . . . . . . 177

Twinning . . . . . . . . . . . . . . . . . . . . . . . . . . . 177

More Speculative Technologies . . . . . . . . . . . . 178

In Vitro Fertilization , . . , , . , , , , . . . . . . . . . 178

Parthenogenesis . . . . . . . . . . . . . . . . . . . , . . 178

Cloning. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178

Cell Fusion . . . . . . . . . . , . . . . . . . . . . . . . . . 179

Chimeras . . . . . . . . . . . . , . . . . . . . . , . . . . . 179

Recombinant DNA and Gene Transfer. . . . . . 179

Genetics and Animal Breeding. . . . . . . . . . . . . . . 179

The National Cooperative Dairy Herd

Improvement Program . . . . . . . . . . . . . . . . . 180

Other Species. . , . . . . . . . . . . . , . . . . . . . . . . . 181

Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . 183

Impacts on Breeding . . . . . . . . . . . . . . . . , . . . 183

Dairy Cattle. . . . . . . . . . . . . . . . . . . . . . . . . . 183

Beef Cattle . . . . . . . . . . . . . . . . . . . . . . . . . . 185

Page

Other Species . . . . . . . . . . . . . . . . . . . . . . . . 187

Other Technologies . . . . . . . . . . . . . . . . . . . . . 188

Aquiculture . . . . . . . . . . . . . . . . . . . . . . . . . 189

Poultry Breeding. . . . . . . . . . . . . . . . . . . . . .189

Issue and Options for Agriculture-Animals . . . . 190

Tables

Table No. Page

30. Heritabilit y Estimates of Some Economically

Important Traits . . . . . . . . . . . . . . . . . . . . . . 169

31. Results of Superovulation in Farm Animals . . 174

32. Experimental Production of Identical

offspring . . . . . . . . . . . . . . . . . . . . . . . . . . 178

33. National Co\v-Year anci A\7erages for All

34

Official Herd Records, by Breed May 1, 1978-

Apr. 30, 1979. . . . . . , . . . . . . . . . . . . . . . . . , 181

P1.e{{i(;te(j Diffc?l’enc[? of klilk Yield of Act i~w

/\l Blllls . . . . . , . . . . . . . . . . . . . . , . . . . . . . 181

Figures

Figure No. Page

30.

31.

32.

33.

34.

Eras in LJ. S. Beef Production . . . . . . . . . . . . 168

Milk Yield/Cow and Cow Population, [Jnit&i

States, 1875 -1975 . . . . . . . . . . . , . . . . . . . . . . 170

Milk Production per Cow in 1958-78 . . . . 17[)

The Way the Reproductive Technologies

Interrelate. , . . . . . . , . . . . . . . . . . . . . . . 175

Change in the Potential Number of ProgcIlj~

per Sire From 1939 to 1979. . . . . . . . . . . . 176
Chapter 9

Advances in Reproductive Biology and

Their Effects on Animal Improvement

Background

During the past 30 years, new technologies

have led to a fundamental shift in the way the

United States produces meat and livestock. One

set of these technologies-the subject of this

section-uses knowledge of the reproductive

process in farm animals to increase production.

‘l-he impacts of existing breeding technologies

have been great, and much progress is still possible through their continued use. The development of new technologies is inevitable as well.

in a market economy like that of the United

States, the factor that most influences the adoption of technology is economics. New technologies in reproductive physiology will be used

widely only if they increase the efficiency of

breeding programs—i.e., only if they provide

greater control over breeding than present

methods do, and only if the economic advantages of the increased control can be recovered. *

But economic factors are not the only ones

that influence technological change—e.g., poultry and livestock production have influenced

and have been influenced by:

● Government regulation such as meat grading standarcls;

. increased awareness of health effects, such

● .\\ (Iis(.ilss(’(1 ill il])j). III-B, \ (’l’)’ (’ill’IJ’ il(lO})lP1’S of il 1(’(’tllloI()~.V

( )1’1(’11 (lo $(1 I 01” ()111(’1’ I Ililll (’(’0110111” ic l’(’il Sl)ll S.

as from the use of antibiotics in livestock

feed;

environmental concerns, such as the problems of waste removal, especially near factory farms;

the growth of knowledge, in-e. g., the reproductive processes of farm animals and

the accuracy of evaluating the genetic

merit of breeding animals; and

complementary technologies such as refrigerated storage and transportation.

New technologies, from breeding to food delivery systems, have reshaped the traditional

American farm into a modern production system that is increasingly specialized, capitalized,

and integrated with off-farm services. Applied

genetics in animal production has been one of

the forces behind these changes. The technologies that have sprung from it include not only

the new, esoteric techniques for cellular manipulation discussed in other parts of this report,

but also more well-known technologies, like artificial insemination. *

“’1’e(.tlll(}l(jgi(>s selected !(w discussion in this part of the report

in~’oh’(? dirf~rl lllillliplllill ion of S(’,X rells. }Iore spwulati\f> twhnologies tol” lllilllil)lllilt ions iit t 11(’ SLlt)(’f>lllllii 1’ 1(’1”~1 il 1’(’ ii SS(>SSP(i }1(>1’[>

iis I\’(?ll. N() (’t’fort t\riis llliid~ [[) (sO\r(’l$ iill te(’hnologies with l)ol~nt iiil

for improving the genetic qualities of Iivestock-e.g., management

techniques like estrus detection and pregnancy diagnosis were

omitted, as wehe various other methods for improving reproduction efficiency.

The scientific era in livestock production

Producing purebred beef livestock has been fenced-in and the longhorn was replaced with

the dominant breeding objective throughout new breeds by the turn of the century—the bemost of the 20th century. The open range of the ginning of the “purebred” era.

American West and Southwest—the “romantic”

era in beef cattle production—lasted until about Pedigree records and visual comparison of

1890. (See figure 30,) Then the range was conformation to breed type were the basic tools

167
168 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Figure 30.— Eras in U.S. Beef Production
Ch. 9—Advances in Reproductive Biology and Their Effects on Animal Improvement ● 169

Table 30.—Heritability Estimates of Some

Economically Important Traits

Trait Heritability

Calving interval (fertility). . . . . . . . . . . . . . . . . . . . . . 10%

Birth weight . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Weaning weight. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

Cow maternal ability . . . . . . . . . . . . . . . . . . . . . . . . . 40

Feedlot gain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

Pasture gain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

Efficiency of gain. . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Final feedlot weight . . . . . . . . . . . . . . . . . . . . . . . . . . 60

Conformation score:

Weaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

Slaughter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Carcass traits:

Carcass grade . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Ribeye area. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70

Tenderness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

Fatthickness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

Retail product (percent) . . . . . . . . . . . . . . . . . . . . 30

Retail product (pounds) . . . . . . . . . . . . . . . . . . . . 65

Susceptibitity to cancer eye . . . . . . . . . . . . . . . . . . . 30

SOURCE: LarryV. Cundiffand KeithE. Gregory, Beet Caft/e Breeding, USDA,

Agriculture Information Bulletin No.2B6, revised November1977, p.9.

may also have economic value,3 but they are

much harder to measure.

The extent to which important economic or

performance traits are genetically determined

and heritable varies from trait to trait and from

animal to animal. (See table 30.) Heritability is

defined as the percentage of the difference

among animals in performance traits passed

from parent to offspring*—e.g~ bulls and

heifers with superior weight at weaning might

average 5 pounds (lb) more than their herdmates. Because weaning weight has an average

heritability estimate of 30 percent, the offspring

of these top performing animals can be expected to average 1.5 1b heavieratweaningthan

their contemporaries (0.30 X 5 = 1.5). This

improvement can normally be expected to be

permanent and cumulative as it is passed onto

the next generation. The improvement accumulates like compound interest in a savings account; gains made in each generation are compounded on the gains of previous generations.

3Nlirhael [. [.erner and H. P. Oonald, &foc/ern Developments in

Anirr?a/ Breeding [New’ }’ork: Academic Press, 1966).

* Heri[ahility and genelir association are important in decisions

iit)oot indi~ idl]iil mat ings. hlost hreeding programs are concerned

~i’ it h spreading geoet ir gain rapidly throughout a population

(Ilt”rd, tlork): thus two other refinements for selection enter the

pirt ur[’-~tvl(>riit ion irlte[’tiil, and selection differential.

Like land, equipment, and cash, breeding

stock represents capital available to the commercial farmer. Because all inputs must be used

efficiently, modern herd or flock managers cannot afford to leave reproduction to chance

mating in the pen or on the range. These pressures for efficient production have been described as follows:4

Where dairymen are judged by the number of

cows milked in an hour, there is no place for the

slow milking cow or the man who will patiently

milk her out. There is no place for the time-consuming hurdle flock of sheep, for the small flock

of chickens maintained under extensive conditions, or for the sow that must be watched

while she farrows. By degrees all classes of

stock are being subjected to selection which

favors animals that need a minimum of individual attention.

The scientific basis for modern breeding has

developed slowly over the last century. Applied

genetics—one part of today’s programs-has

helped modernize livestock and poultry breeding by elaborating on the variation of continuously distributed traits in a population; carrying

over what was known about rapidly reproducing laboratory species, like fruit flies or mice, to

the much slower reproduction of large farm

animals; and developing the statistical techniques for predicting breeding values or merit

and analyzing breeding programs. s

Two examples show the power of breeding

tools and the increased efficiency and productivity of today’s breeders’ stocks.

● Over the past 30 years, the average milk

yield of cows in the United States has more

than doubled. At the same time, the number of dairy cows in the United States has

been reduced by more than 50 percent.

(See figure 31.) Of this increase in output

and efficiency, more than one-fourth can

be attributed to permanent genetic change

for at least one breed (Holsteins) participating in the Dairy Herd Improvement Program. (See figure 32.)

● Poultry production in the United States has

become the most intensive industry among

qlhid., p. 20.

~lbid., p. 126.
170 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Figure 31 .—Milk Yield/Cow and Cow Population,

United States, 1875-1975 -

1875 ’85 ’95 ’05 ’15 ’25 ’35 ’45 ’55 ’65 1975

Year

SOURCE: J. T. Reid, “Progress in Dairy Cattle Production,” Agricultural and

Food Chemistry. Past, Present, and Future, R. Teranishi (cd.)

(Westport, Corm.: Avi Press, 1978).

Figure 32.—Milk Production per Cow (Holsteins) in

1958=78 (New York and New England)

2-year old Holstein cows in DHIA by Al. Sires

+ 4000

+ 3000

+ 2000

+ 1000

Base

1958 1962 1966 1970 1974 1978

Year

SOURCE: R. H. Foote, Department of Animal Science, Cornell University,

Ithaca, N.Y. from unpublished data of R. W. Everett, Corneil

University.

those for farm species. For turkeys, the use

of Al in breeding for breast meat has been

so successful that commercial turkeys can

no longer breed naturally. The bigbreasted male, even when inclined to do so

finds it physically impossible to mount the

female. As a result, a full 100 percent of the

commercial turkey flock in the United

States is replaced each year using Al. In

other species of poultry as well, production

processes have become equally efficient.

As A. W. Nordskog has noted:

Compared with the breeding of other economically important animals, poultry breeding has been the first to leave the farm . . . to

become part of a sophisticated breeding industry. On a commercial level, chickens have

been the first to be commercially exploited

by the application of inbreeding-hybridization techniques, as earlier used in corn, as

well as by methods of selective improvement

using the principles of quantitative genetics.

Thus, the poultry industry, compared to

other animal industries, seems to have been

the quickest to apply modern methods of

genetic improvement, including the employment of formally trained geneticists to handle

breeding technology plus the use of computers and other modern business methods.6

Scientific production

Farm resources include land, labor, capital,

and, increasingly, new knowledge. Today, those

who innovate recapture the costs of innovating

by maintaining output while lowering costs or

by increasing output while holding costs down.

Some results of the drive toward efficiency have

included included increasing specializat ion, intensified

use of capital and land relative to labor and integration of production phases .

Poultry and livestock operations have slowly

become specialized over the past 50 years. The

farmer who used to do his own breeding, raising, feeding and slaughtering is disappearing.

Now, the beef cattle industry in the United

States consists of: the purebred breeder who

provides breeding stock, the commercial producer, the feeder, the packer, and the retailer.

Similar specialization has occurred for most

other species—e.g., less than 15 primary breeders maintain the breeding stock that produces

the 3.7 billion chickens consumed each year in

the United States. The emergence of other specialized services—such as AI providers, manage-

‘A. W. Nordskog, “Success and Failure of Quantitative Genetic

Theory in Poultry” in Prwceedinga of the International Conference

on Quantitative Genetics, Edward Pollacket, et al. (cd.) (Amers,

Iowa: Iowa State University Press, 1977), pp. 47-51.
Ch. 9—Advances in Reproductive Biology and The Effects on Animal lmprovement ● 171

ment consultants, equipment manufacturers—

has accelerated the trend toward specialization,

and has given the commercial operator more

time to concentrate on his specific contribution

to the chain of production.

Intensification is the increasing use of some

inputs to production in comparison to others.

Increasing the use of land and capital relative to

labor describes the development of U.S. agriculture, including livestock raising, in this century.

The “factory” farm typifies this trend. Herds

and flocks are bred, . born, and raised in enclosed areas, never seeing a barnyard or the

open range. The best examples of land- and capital-intensive systems are those of poultry

(layers, broilers, and turkeys), confined hog production, drylot dairy farming, and some veal

production.

The greater use of land has been encouraged

by several factors, including improved corn production for confined hog feeding, programs of

preventive medicine curtailing the spread of

diseases in close spaces, and environmental control (light, temperature, water, humidity) to increase output under closely controlled conditions. However, extensive ranching for beef and

sheep is still common in the United States; the

difficulties associated with detecting estrus

(“heat”) in these species and their relatively slow

rates of reproduction have made it uneconomical to invest in them the capital necessary for

intensive farming. Furthermore, beef and sheep

on extensive systems forage on marginal land

that might otherwise have no use. Beeflot feeding, or the fattening of cattle before slaughter at

a centralized location, is the only aspect of the

beef industry that is land-intensive; in 1977, approximately one-fourth of U.S. beef cattle were

fed "

7

Linking phases of production to eliminate

waste or inefficiencies in the system has progressed with great speed. For some species,

such linkages now extend from breeding to the

supermarket (and, in the case of fast food

chains, to the dinner table). Integration includes

7Lyle P. Schertz, et al., Another Revolution in U.S. Farming?

USDA, ESCS, Agricultural Economic Report No. 441, December

1979.

the linking of supply industries (feeds, medicines, breeding stock) with production and then

with marketing services (slaughtering, dressing,

packaging). Entire industries and the Government in combination have produced a complex

chain of operations that makes use of Government inspectors, the pharmaceutical industry,

equipment manufacturers, the transportation

industry, and the processed feed industry in addition to the traditional commercial farmer.

Because of this complex linkage, meat grades,

cuts, and packaging have become fairly standard in the American supermarket. Shoppers

have come to expect these standards; consumers wanting special services have learned to pay

more for them. Thus, the American farm has

changed radically over the past 30 years. This

change has been described as follows:8

As farming enterprises grow larger, their

management have to equip themselves with information and resort to technologists to help

them reach decisions and plan for more distant

goals. Industrial developments of this kind

widen the range of farming activities, since the

old style farmer, sensitive to local markets and

operating on hunches, remains as a contrast to

those for whom farming is rapidly becoming

more of a programme than a way of life.

Resistance to change

New technologies in U.S. agriculture and new

ways of producing food and fiber have been

both a cause and an effect of the movement

from farms to cities in the 20th century. Commercial farmers, operating on thin or nonexistent profits and under extreme competition,

have had strong reason to innovate. They have

been forced by the availability of new technologies either to do so or to watch their potential

earnings go to the neighboring farmer. Various

policies that have been adopted to soften the impacts of the “technological treadmill,” have

somewhat slowed the exodus from the farms.

They may have been adopted for social reasons,

but they have also become increasingly costly to

society. The taxpayer pays for them; the consumer pays as well for every failure to innovate

on the farms.

Wundiff, et al., op. cit., p. 9.
172 ● Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

Besides a lack of capital or a lack of interest in

innovating, some farmers have resisted applied

genetics because efficiency is not their most important priority. This attitude has been described as follows:

It is easy to see why breeders are unreceptive

to the science of genetics. The business of

breeding pedigree stock for sale is not just a

matter of heredity, perhaps not even predominantly so. The devoted grooming, feeding and

fitting, the propaganda about pedigrees and

wins at fairs and shows, the dramatics of the

auction ring, the trivialities of breed characters,

and the good company of fellow breeders, constitute a vocation, not a genetic enterprise.

Farmers are traditionally an independent

group. Many believe that they may not directly

recapture the benefits of participating in a

breeding program based on genetics; having no

records on one’s animals is often preferable to

discovering proof that one’s herd is performing

poorly. On the other hand, one impact of AI has

been to demonstrate to farmers the value of

adopting new technologies. Furthermore, the

economic reward of production records has increased, since AI organizations purchase only

dairy sires with extensive records on relatives.

Somefuture trends

Applied genetics in poultry and livestock

breeding comprise a group of powerful technologies that have already strongly influenced

prices and profits. Nevertheless, the effect of

genetics is only just beginning to be felt; much

improvement remains to be made in all species.

It has been observed that modern genetics:l0

. . . provides a verifiable starting point for the

development of the complex breeding operation

that many populations now require . . . (which)

are as far removed from simple selection as the

motor car is from the bicycle.

Of these technologies, some are already in

regular use, some are in the process of being applied, and others must await further research

and development before they become generally

available.

Slbid., p. 170.

‘“E. P. Cunningham, “Currem Developments in the Cenetics of

[,iwstock Improvement,” in 15th International Conference on Animal Blood (hwups and Biochemistry, Genelics 7:191, 1976.

Societal pressures are one of the many factors that influence the introduction of these

technologies. Several developments around the

world will have a clear impact on innovation in

general and on genetics in particular:

An expanding population, with its growing

demand for food products of all kinds.

The growth in income for parts of the population, which may increase the demand

for sources of meat protein.

Increasing competition for the consumer’s

dollar among various sources of protein,

which could reduce demand for meat.

Increasing competition for prime agricultural land among agricultural, urban,

and industrial interests. Less-than-prime

land may also be brought back into production as demand rises, and the same pressures may cause land prices to rise high .

enough to encourage greater, or intensified, use of land in livestock production.

Increasing demand for U.S. food and fiber

products ‘from abroad, leading to opportunities for increased profits for successful

producers.

Changes like these will strongly affect the

way American farmers produce food and fiber

products. The economics of efficiency and a

growing world population will continue to place

pressure on the agricultural sector to innovate.

In animals and animal products, efficiencies will

be found in all steps of production. Efforts will

be made to increase the number of live births

and to reduce neonatal calf fertility, presently

one of the costliest steps—in terms of animals

lost–throughout the world. Estimates of the potential monetary benefits of the application of

knowledge obtained from prior research in reproductive physiology range as high as $1 billion per year. Another area for great economies

in production is genetic gain. Much genetic

progress remains to be made in all species.

Certain technologies promise to increase the

ability of farmers to capitalize on the genetic improvement of economically important traits.

Suppliers of genetic material (semen, embryos)

will focus increased attention on the value of

their products for sale both in the United States

and abroad.
Ch. 9—Advances in Reproductive Biology and Their Effects on Animal improvement ● 173

The development and application of certain

key technologies will affect related technologies—e.g., the availability of reliable estrus

detection and estrus synchronization methods

should increase the use of AI and embryo transfer in beef and dairy cattle, thereby spreading

genetic advantage. Further progress in the

freezing of embryos should facilitate the genetic

evaluation of cows and heifers.

research funds, changing consumer tastes, and

growth of regulations (for instance, stricter controls on environmental quality or hormonal

treatments). The expansion of an animal rights

movement may influence the degree to which

confinement housing, and therefore controlled

breeding, is acceptable. And increased energy

costs may either encourage development of the

technologies (through efforts for greater effiOther trends that may influence techno- ciency) or discourage them (through greater use

logical change include the shifting availability of of forage and extensive systems).

Technologies

Sexual reproduction is a game of chance. Because sperm and ova each contain only a random half of the genes of each parent, the number of possible combinations that can result is

nearly infinite. Some progeny are likely to survive and reproduce; others die either before

birth or without producing offspring.

The great variation achieved through sexual

reproduction produces certain animals that

satisfy the needs and desires of the breeder far

more than others. On the other hand, the offspring of these outstanding animals are usually

less so than their parents, although they are

generally still above average.

Animal breeders have invested great effort in

improving succeeding generations of domestic

animals, both by limiting the differences due to

the chance associated with sexual reproduction

and by taking advantage of the favorable combinations that occur. Examples of these efforts include keeping records, establishing progeny

testing schemes, amplifying the reproduction of

outstanding individuals by AI and embryo transfer, and establishing inbred lines to capitalize on

their more reliable ability to transmit characteristics to their offspring.

Because of these efforts, and because dairy

cattle breeders have adopted innovative technologies through the years, far more is known

about reproduction in the cow than in other

farm animals. The demand for milk and beef

has provided an impetus for the speedy introduction of technologies that might prove economically advantageous.

Several observations can be made about the

state of the art for 16 technologies that enhance

the inherited traits of animals. (See also app.

II-C.)

The technologies are at different stages of research and development.

The practice of AI in dairy cattle has had the

greatest practical impact of all the genetic technologies used in the breeding of mammals. In

contrast, not a single farm animal has been successfully raised after a combination of in vitro

fertilization and embryo transplant. The usefulness of several of the technologies for animal

production, such as recombinant DNA (rDNA)

and nuclear transplantation, is purely speculative at this writing.

The usefulness of the technologies differs from

species to species. .

These differences can often be explained by

biological factors-e. g., sperm storage capabilities are currently limited for swine because

freezing kills so many of the sperm. Management techniques are important as well; extensive beef-raising systems have in the past made

estrus detection and synchronization impractical, thereby limiting the use of AL (Fewer than

5 percent of the U.S. beef herd are artificially inseminated, compared with 60 percent of the na-
174 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

tional dairy herd.) And economics can also play

a role; in general, the lower an animal’s value,

the less practical the investment in the technologies, some of which are relatively expensive.

Several technologies are critical to the introduction

of others.

A methodology that could reliably induce

estrus synchronization increases the economic

feasibility of AI and embryo transfer. Likewise,

the refinement of embryo storage and other

freezing techniques would advance the development of those technologies still being developed,

like sex selection and embryo transfer. Advances in in vitro fertilization will be especially

useful to a better understanding of basic reproductive processes and therefore to the development and application of the more speculative

technologies.

The technologies interrelate.

All the technologies combined make possible

almost total control of the reproductive process

of the farm animal: a cow embryo donor may be

superovulated and artificially inseminated with

stored, frozen sperm; the embryos may be recovered, then stored frozen or transferred directly to several recipient cows whose estrous

cycles have been synchronized with that of the

donor to insure continued embryonic development. Before the transfer, a few cells may be

taken for identification of male or female chromosomes as a basis for sex selection. Finally,

two embryos may be transferred to each recipient in an effort to obtain twins. (See figure 33.)

Techniques not yet commercially applicable

all require embryo transfer in order to be useful. They include in vitro fertilization, parthenogenesis, production of identical twins, cloning,

cell fusion, chimeras, and rDNA technology.

The technologies described in this section are

designed to increase the reproductive efficiency

of farm animals, to improve their genetic merit,

and to enhance general knowledge of the reproductive process for a variety of reasons, including concern with specific human medical problems, such as fertility regulation and better

treatments for infertility.

Technologies that are presently useful

SPERM STORAGE

The sperm of most cattle can be frozen to

– 196° C, stored for an indefinite period, and

then used in in vivo fertilization. Although

many of the sperm are killed during freezing,

success rates [or successful conceptions (table

31)] combined with other advantages of the

technologies are enough to ensure widespread

use of the technology. Short-term sperm storage

(for one day or so) is also well-developed and

widely used.

The major advantages of storing sperm are

the increased use of desirable sires in breeding

(see figure 34), the ease of transport and spread

of desirable germplasm throughout the country

and the world, and the savings from slaughtering the bull after enough sperm has been collected. The sperm can also be tested for venereal and other diseases before it is used. Therefore, the use of sperm banks is expected to increase. Little change is anticipated in semen

processing, other than the continued refinement of freezing protocols, which differ for

each species.

ARTIFICIAL INSEMINATION

The manual placement of sperm into the

uterus has played a central role in the dissemination of valuable germplasm throughout the

world’s herds and flocks. Virtually all farm species can be artificially inseminated, although use

of the technology varies widely for different

species—e.g., 100 percent of the Nation’s domestic turkeys are produced via AI compared with

less than 5 percent of beef cattle. Even honeyTable 31.—Results of Superovulation in

Farm Animals

Average number

ovulations normally Number of ovulations

expected with superovulation

cow . . . . . . . . . 1 6-8

Sheep. . . . . . . . 1.5 9-11

Goat. . . . . . . . . 1.5

Pig . . . . . . . . . . 13 30

Horse. . . . . . . . 1 1

SOURCE: George Seidel, Animal Reproduction Laboratory, Colorado State University, Fort Collins, Colo.
Ch. 9—Advances in Reproductive Biology and Their Effects on Animal Improvement ● 175

Figure 33.

Bull

—The Way the

Sperm

Reproductive Technologies Interrelate

Artificial

insemination

Superovulated

cow

Sexed?

Frozen?

Calves

Recovered

embryos

Sexed?

Recipient herd: synchronized estrus

Embryo transfer

Each get two for twinning

Photo Credit: Science

These 10 calves from Colorado State University were the

result of superovulation, in vitro culture, and transfer to

the surrogate mother cows on the left. The genetic

mother of all 10 calves is at upper right

SOURCE: Office of Technology Assessment
176 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Figure 34.—Change in the Potential Number of

Progeny per Sire per Year From 1939 to 1979

Year

SOURCE: R. H. Foote, Department of Animal Science, Cornell University,

Ithaca, N. Y., unpublished data.

bees and fish can now be artificially inseminated.

It permits the widespread use of germplasm

from genetically superior sires. It saves the

farmer the cost of maintaining his own sires and

is valuable in disease control, especially when

germplasm, rather than animals, is imported or

exported. An important barrier to the wider use

of Al, especially in producing beef cattle, is the

need for application of reliable estrus detection

and estrus synchronization technologies.

An expanded role for Al in the future will depend on the availability of accurate information

about the genetic value of sperm available for

insemination. A nationwide information system

for evaluating germplasm presently exists for

only one species, dairy cattle.

ESTRUS SYNCHRONIZATION

Estrus, or “heat,” is the period during which

the female will allow the male to mate with her.

The synchronization of estrus in a herd, using

various drug treatments, greatly enhances Al

and other reproduction programs.

Federal regulations that limit the use of prostaglandins 01* pro#?sto#?ns to inc]uce s~nchronized estrus in horses and nonlactating cows

are the ma jot’ barrier to more 1%’idespread use of

exist ing technology.

SUPEROVULATION

SL1~]el*()\~llliitioll is the hormonal stimulation of

the female, resulting in the release from the

Okr; it’y of a kit>ger number of ova than nornla].

(St?e tiil]]e 31. ) (k)mhined with AI and WNIII*YO

transt’er of’ the i“(?rtilized ok’;i

mothers, sL]l)eI*ol’l]liite(i ova c;iI

production of normal of fsl)ring

rates of succc?ss as thos(? toilowi

kit ion.

into surro#ite

I result in th e

with the sanle

)g Ilol”lllal o\’u-

‘1’he greatest Imrrier to sll~ei’(]il]l:itiotl is that

the degree of success cannot be predicted for an

inciividual aninuil. other barriers incluc]e widely

Varvillg q~l;~]itv of hormone batches fOI’ Ol’Ul~-

tiot; treatmeni, l~()()d and Drug Administration

(FDA) restrictions, and lack of ckita from which

to judge the effects of repeated sllpel’ot~t]llltioll.

In the future, increaseci L]tl(le].stiillciitlg of

t)asic physiological mechanisms will facilitate et’-

forts to imprm~e the technology. It has additional commercial potential for sheep and cattle

hustmnclry, and much current effort is directed

tmvarcls det~eloping and testing a commercial

procedure.

EMBRYO RECOVERY

The ability to collect fertilized ova from the

oviducts or uterus is a necessary step for emt)ryo transfer or storage and for many experiments in reproductive biology. The technology

is especiall.v important for research into producing identical twins, performing embryo biopsies

for sex (ietel*llli]]~itiotl, and other projects. ConlI)ining superovulation, art i ficial insemination,

and embryo rec(n’ery makes it possihle to collect embryos from a young heifer before reaching puberty. When some disorder has damaged

the oviducts or uterus, embryo recovery from a

Va]uilt)]e l~ninlal makes procreation possible.

Both surgical and nonsurgical methods are

currentl.v in use. Surgical recovery is necessary

for sheep, goats, and pigs; such operations are

limited by the development of scar tissue. Non-
Ch. 9—Advances in Reproductive Biology and Their Effects on Animal improvement ● 177

surgical embryo recovery is preferred for the

cow and the single ovulation of the horse. The

approach is especially important in dairy cattle,

since it can be performed on the farm without

interrupting milk production.

No significant advances can be predicted for

the immediate future.

EMBRYO TRANSFER

Embryos can be removed from one animal

and implanted into the oviduct or uterus of

another. Both surgical and nonsurgical methods

are currently in use, though success rates of the

latter are much lower.

The technology can obtain offspring from females unable to support a pregnancy, increasing the number of offspring from valuable females and introducing new genes into pathogen-free herds. Because more offspring can be

obtained from the donor, undesirable recessive

traits can be rapidly detected. The technology is

also used, along with short- or long-term storage

of the embryos, as a means of transporting

germplasm rather than the whole animal. Current barriers to its further use are the costs in

personnel and equipment, especially for surgical procedures, and the provision of suitable

recipients for a successful transfer.

The use of embryo transfer should increase

in the future, especially with animals of high

value. Nonsurgical methods will increasingly

replace surgical ones, especially for cows and

horses. A role for embryo transfer can also be

predicted in progeny testing of females, obtaining twins in beef cows, obtaining progeny from

prepubertal females, and in combination with

in vitro fertilization and a variety of manipulative treatments (production of identical twins,

selfing or combining ova from the same animal,

genetic engineering).

EMBRYO STORAGE

The ability to store embryos increases the

advantages of embryo transfer procedures, lowers the cost of transporting animal germplasm,

and reduces the need to synchronize estrus in

recipients. It will also be important in the study

and control of genetic drift in animals.

Adequate culture systems exist for short-term

storage of embryos. They hai’~? been d[?~~(?kq)ed

by tl*ial-iil]d-el.l’01* and are not optimally defin[?d

for farm species at present. Nmfertheless, COJ%r

embryos ha~’e been stored for 3 cki.vs in the tied

oviduct of a I’al)l]it.

Long-term storage, or freezing of enlbr.yes,

exists, but protoco]s need to be improkfed. As

many as two-thirds of the stored embryos die

with present methods. However, for some uses

embryo freezing is already profitable.

In the future, the de~’elopnwnt of precise enlbr~o culture technology tvoL]ld help the deJwlopment of all technologies in~~olkting the prolonged manipulation of gametes and embryos

outside the reproducti~w tract. Event uall.v, as

freezing technology imprm’es, nearly all embryos taken from cattle in North America will

be stored, rather than transferred inln]ediately.

It appears that embryos successfully stored ~vi]l

survive for several centuries and possibly for

millenia.

SEX SELECTION

The ability to determine the sex of the Llnborn, or of sperm at t’ertilization, will ha\’e numerous practical and experimental applications.

The most reliable method is karyotyping, by

means of which nearly tm’o-thirds of embryos

can be sexed. Another method, which tries to

identif~v sex-specific products of certain genes,

is under development. A reliable method for

separating ma]e-producing sperm from femaleproducing sperm has not been achieved, though

several patents are held on ~’arious tests of this

type.

Before any method has any practical effect on

the production of farm animals it must become

simple, fast, inexpensive, re]iable, and harmless

to the embryo. The present state of the art is

largely a consequence of research in male fertility and in sperm survival after frozen storage.

TWINNING

Twins can be artificially induced by using

either embryo transfer or hormonal treatments.

The first approach is more effective. Selection

among female sheep for natural twin produc-
178 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

tion has been very rewarding, while selection

for twinning in other species has not received

much attention.

Twinning in nonlitter-bearing species would

greatly improve the feed conversion ratio of

producing an extra offspring. The most important barriers, besides the high cost of embryo

transfer techniques, include extra attention

needed for the dam during gestation, parturition, and lactation.

More speculative technologies

IN VITRO FERTILIZATION

The manual joining of egg and sperm outside

the reproductive tract has, for some species,

been followed by successful development of the

embryo through gestation to birth. The species

include, at this writing, the rabbit, mouse, rat,

and human. Consistent and repeatable success

with in vitro fertilization in farm species has not

yet been accomplished. The cases of reported

success of in vitro fertilization, embryo reimplantation, and normal development in man are

beginning to be documented in the scientific

literature.

The in vitro work to date has attempted to develop a research tool so that the physiological

and biochemical events of fertilization could be

better understood. Despite the wide public attention it has received in the recent past, the

technology is not perfected and will have little I

I practical, commercial effect in producing individuals of any species in the near future.

Practical applications would include: a means

of assessing the fertility of ovum and sperm; a

means of overcoming female infertility by embryo transfer into a recipient animal; and, when

coupled with storage and transfer, a means of

facilitating the union of specific ova and sperm

for production of individual animals with predicted characteristics.

Many of the practical applications should become available within the next 10 to 20 years.

Further development, along with the storage of

gametes, should allow fertilization of desired

crosses. This technology may be combined with

genetic engineering and sperm sexing in the

more distant future.

PARTHENOGENESIS

Parthenogenesis, or “virgin birth, ” is the initiation of development in the absence of sperm.

It has not been demonstrated or described for

mammalian species, and the best available information indicates that the maintenance of parthenogenetic development to produce normal

offspring in mammals is presently impossible.

CLONING

The possibility of producing genetically identical individuals has fascinated both scientists

and the general public. As far as livestock are

concerned, there are several ways to obtain

genetically identical animals. The natural way is

through identical twins, although these are rare

in species other than cattle, sheep, and primates. For practical purposes, highly inbred

lines of some mammals are already considered

genetically identical; first generation crosses of

these lines are also considered genetically identical and do not suffer from the depressive effect of inbreeding.

Laboratory methods for producing clones include dividing early embryos. The results of recent experiments in the production of identical

offspring using these techniques are shown in

table 32.

Another methodology involves the insertion

of the nucleus of one cell into another, either

before or after the original genetic complement

of the “receiver” cell is destroyed. Researchers

have found in certain amphibia that nuclear

transplantation from a body cell of an embryo

into a zygote can lead to the development of a

sexually mature frog.

Table 32.—Experimental Production

of Identical Offspring

Methodology Result

Dividing 2-cell embryo in 1 pair identical mouse twins

half

Dividing morulaea in half 8 pairs of identical mouse

twins

Dividing 2-cell embryos in 5 pairs of identical sheep

half twins

Dividing 4-cell embryos in 1 set identical sheep

four parts quadruplets

aAn embryo with 16 to 50 cells; resembles a mulberry.

SOURCE: Benjamin G. Brackett, School of Veterinary Medicine, University of

Pennsylvania, Kennett Square, Pa.
Ch. 9—Advances in Reproductive Biology and Their Effects on Animal improvement . 179

The ideal technique for making genetic copies

of any given adult mammal involves inserting

the nucleus from a body cell (not a sex cell) from

an adult individual into an ovum. Achieving this

will probably take years, if indeed it is possible

at all, since there is some evidence that most

adult body cells are irreversibly differentiated. *

Serious technical barriers must be overcome

before advantages in animal production can be

foreseen.

CELL FUSION

This technology fuses two mature ova or fertilizes one ovum with another. Combining ova

from the same animal is called “selfing.” The

combination of ova has resulted in very early

development of the transferred embryo, but no

further development has been reported.

Cell fusion technology may someday prove

useful for transferring genetic material from a

somatic cell into a fertilized single-cell embryo

for the purpose of cloning. Selfing would rapidly result in pure genetic (inbred) lines for use as

breeding stocks. The technique could also lead

to the rapid identification of undesirable recessive traits that could be eliminated from the

species.

CHIMERAS

The production of chimeras requires the fusion of two or more early embryos or the addi-

● Ii) JanLIary 1981, it was reported that body cells from a very

[~iii’l,v embryo could art as donors of nuclei for cloned mice.

tion of extra cells to blastocysts. These genetic

components may be from closely related but different species.

Live chimeras between two species of mouse

have been produced. However, practical applications of chimera technology to livestock are

not obvious at this stage of development. The

main objective of this research is to provide a

genetic tool for a better understanding of development and maternal-fetal interactions.

RECOMBINANT DNA AND GENE TRANSFER

The mechanics of directly manipulating the

DNA molecules of farm animals have not yet

been worked out. However, cells from mice

have been mixed with pieces of chromosomal

DNA, which became stably associated with the

cells’ own DNA. In addition, on September 3,

1980, the successful introduction of foreign

DNA into mouse embryos was announced. The

embryos were implanted into surrogate mothers who gave birth to mice containing altered

DNA. whether or not the DNA was active is unknown at this writing.

Knowledge of the genetics of farm animals

must improve before rDNA or other gene transfer methods will be of practical benefit in

producing meat and livestock products. Before

genes can be altered they must be identified,

and gene loci on chromosomes must be

mapped. Work toward this goal has begun only

recently and rapid progress cannot be anticipated. Multivariate genetic determinants of

characteristics are anticipated to be the rule.

Genetics and animal breeding

Two characteristics distinguish the reproduction of farm animals from that of single-cell organisms: animal reproduction is sexual–male

and female germ cells must be brought together

to initiate pregnancy and produce offspring;

and animal reproduction is slower (the generation interval is longer), thus the economic benefits of specific gene lines may take years to be

captured. These two characteristics limit the

speed and extent to which genetic improvements can be made. Reliable information about

the genetic value of particular individuals is the

key to overcoming limitations, for it can simplify specific breeding decisions and spread desirable genes throughout the Nations’s herds and

flocks.

The use of applied genetics for farm species is

indirect. Breeders do not work with individual

genes; rather, they must accept a genetic pack-
180 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

age that inclucies both beneficial and harmful

tl’iIitS. i‘ ‘W? bl’eeck?r’s most illlpOI’tiiIlt capital i s

(?mt)odied in the iininliils with which he works.

‘J-o Llf)~]*iid(? this (;iij)itiil, to ii~creiise the genetic

\’iill]~ ot his br[?eding stock, the breeder must

Iliit’t? N?liiiblC? illfol’miitioll” 011 the genetic value

of the ~(?l’ll~[)liis]ll he is considering introducing.

Since iiil incli\’icltiiil tiirnlel. usuiill~ does not

tlii\e the t’esou]x:es to collect iilld process data

on ~]ert’ol*illiill(~e of indi\’iduiils outside his own

herds, he must turn to outside sources of inforllli~tioll when deciding which ne\v germplasm to

int reduce.

‘1’he requirements of such iill information system ii]’(? [?xtensitw. Ill the [Jnited States today,

onl.v om? such system exists. The National Co-

()}]~riitiir[? t)iiir~ HeI*ci Impro\rement Progra m

(N(II)HIP) is ii illoclel piwgraNl thi~t could b e

ii(~iipt E?(l to Ot IIC?l’ species where the benefits

from ii(l\’iiil[~~(l technok)gies ~’ould be enhanced

l)y at’;liliit)ility of populiit ioflwide data.

The NationaZ Cooperative Dairy Herd

Improvement Program

Over the past 50 years, the U.S. dairy industry has used test records of individual animals

to help in breeding decisions. NCDHIP is a nationwide program for collecting, analyzing, and

disseminating information on the performance

of dairy cattle.12 It is the result of a memorandum of understanding among Federal and State

agencies, local dairymen, and industry groups

across the United States.

In NCDHIP, local Dairy Herd Improvement

Association (DHIA) officials go to the dairies to

collect the performance data on individual animals. These data then become part of the @~icial Dairy Recordkeeping Plans. The data are

standard for all participating herds across the

United States. They are sent to the Animal Improvement Programs Laboratory (AIPL) at

USDA in Beltsvi]le, Md., which analyzes them

and incorporates them into the “USDA-DHIA

11 I%il ip Ililll(ll(’1’, i3iologv and the P“uture qf- Man (New York: OxI’ord t lni~(}rsit}’ Press, 1 $)70), pp. .5.5.5 -557.

IZI:O1. il [.oll)PI(~l(! llistort~ of ~](?l’tol’llliitl(:e tesi ing of dairy cattle

in the [ Inil(’({ Stii((~s, sw: ( ;t~l.iild J. King, The Nationa/ Cooperative

/)airy Herd Improtwnent Program, [liiir}~ Herd Improvement 1~ttt>r 49, N(), 4, July 1973, IISI)A, /\ KS.

Sire Summary List, ” published biannually.

These summaries are public information.

In addition to the official plan, NCDHIP also

includes several unofficial plans, which have

less stringent regulations for data collection but

which offer each dairyman a comparison of his

herds with other herds across the Nation. The

results of unofficial plans are not intended to be

used as guidelines for selecting germplasm from

outside one’s herd.

The following characteristics contribute to

NCDHIP’s success:

●

●

●

It is a cooperative program; no group or individual is forced to participate. Nevertheless, it has successfully brought together

individuals, State and Federal agencies,

breed associations, and professional and

scientific societies for the pursuit of a common goal. It is almost totally financed by

the dairymen themselves. In the national

coordinating group, all those with an interest in the industry have a voice in formulating policy for the program.

It is flexible; a dairyman can use the performance records from the unofficial plans

to evaluate the animals within his herd or

he can turn to the official sire summaries

to make comparisons with participating

herds throughout the Nation. These data

are useful both for comparing the performance of one’s herd and breed with others

and for selecting new germplasm for introduction into the herd.

Its data are regarded as impartial; disinterest

on the part o-f the local DHIA official who

collects the data and the high security surrounding the processed information are

central to the program’s success. AIPL’s

analyses and sire summaries are respected

both nationally and internationally, in no

small part because of freedom from commercial pressures.

Approximately 36,000 herds with almost 2.8

million cows were enrolled in the official plans

of NCDHIP in 1979. In each of 18 years recorded

between 1961 and 1978, cows enrolled in the

Official Dairy Recordkeeping Plans in NCDHIP

have outproduced cows not enrolled by over
Ch. 9—Advances in Reproductive Biology and Their Effects on Animal improvement ● 181

4,000 lb of milk per lactation. In the testing year

(1977-78), the superiority surpassed 5,000 lb per

cow. This 5,()()()-lb superiority represents 52

percent more milk per lactation. The increases

in production per cow result from improvement

in both management techniques and genetic

producing ability.

Several factors influence the rates of participation in the NCDHIP from State to State, from

region to region, and from breed to breed. In

some States, expansion of NCDHIP membership

is not a high priority of the State Cooperative

Extension Service. In some areas, the relative

importance of dairying as an enterprise is low;

therefore, a strong local DHIA organization

does not exist. Likewise, in areas where dairying is a part-time operation, dairymen have less

time and initiative for participating in the program (although many participate in NCDHIP’s

unofficial plans). Where dairymen rely on their

own bulls and use little AI in breeding, progeny

testing is extremely limited. No single factor

causes dairymen in some States to take greater

advantage of the superior germplasm available

to them. The importance of strong national

leadership cannot be overemphasized in explaining the great differences among breeds in

participation rates. (See table 33.) Farsighted

leadership played a large role in developing the

genetic gain of Holsteins, which represent 90

percent of the U.S. dairy herd today.

The genetic gains resulting from NCDHIP are

impressive, suggesting a model for spreading

genetic superiority throughout the Nation’s

other herds. NCDHIP also shows the importance

Table 33.-National Cow-Year and Averages for

All Official Herd Records, by Breed

May 1, 1978-Apr. 30,1979

Cow-years

lreed (#) Milk (lb) Fat (VO) Fat (lb)

~yrshire . . . . . . . . 17,135 11,839 3.96’XO 469

iuernsey . . . . . . . 57,577 10,858 4.64 504

Iolstein . . . . . . . . 2,297,684 15,014 3.64 547

ersey. . . . . . . . . . 89,449 10,231 4.90 501

rown swiss. . . . . 24,247 12,368 4.04 500

Iilking shorthorn 2,130 10,451 3.65 381

Iixed and others. 83,139 13,077 3.80 497

)URCE: U.S. Department of Agriculture, Science and Education Administration, Dairy Herd hnprovement Letter 55, #2, December 1979, pp. 5-6.

of combining reliable evaluation of germplasm

with the use of reproductive technologies.

These technologies are of only academic interest when they are used alone; it is when

superior germplasm can be spread throughout

the Nation that the American consumer

benefits.

Other species

Progeny testing schemes for other species are

not as developed as they are for dairy cattle.

There are several reasons for this lack of

testing:

●

●

●

●

Difficulty in establishing a selection objective

around which to design a testing program.

Milk yield and fat content were obvious

traits for selection in dairy cattle. other

species have no such simple traits for selection. It has been observed that, “The lack of

definition of economic selection objectives

in a precise, soundly based manner is one

of the serious weaknesses of much animal

breeding of the past. ”13

Differences in management systems. Artificial insemination is essential to the introduction of superior germplasm; where it is

difficult to practice Al, elaborate testing

schemes are not useful—e.g., in the Nation’s beef herds, progeny testing will have

to await more widespread use of AI.

Though swine are increasingly raised in

confined housing systems, poor fertility of

boar sperm after freezing and thawing and

heat detection difficulties have limited the

use of AI.

Conflicting commercial interests. Beef bulls,

for example, continue to be sold to some

extent on the basis of fancy pedigrees and

lines, with relatively little objective information on their genetic merit. Although

some genetic improvement programs now

exist, the beef breed associations may not

support interbreed comparisons because

some breeds would show up poorly.

Conflicts between short- and long-term gains.

Cross-breeding for the benefits of hybrid-

‘31.. E. A. Rowson, ‘Techniques of l.ivestoch lnlpro~renm~t ,“ Ou[-

/ook on Agricuhure 6:108, 1970.

76-565 0 - 81 - 13
182 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

ization is particularly attractive to owners

of commercial herds and flocks who constantly replace their stocks. This genetic

improvement is noncumulative—the improvement does not continue from generation to generation. At present, no strong

interest exists for improving the Nation’s

beef herd as a whole, and the individual

breeder cannot effectively evaluate the

germplasm available to him.

Swine .—There is no Nationwide testing program for hogs in the United States. * However, a

study of needed research prepared by the

USDA in 1976 noted that the production rate of

approximately 13 pigs marketed per sow per

year in the United States could be significantly

improved. The biological potential is at least 20

to 25 pigs per year. Similarly, a successful

breeding program, along with other managerial

changes, could reduce the fat and increase the

lean content of pork by as much as 10 to 15 lb

per carcass.

The ARS study noted that “. . . an area that

warrants particular attention is the development of a comprehensive national swine testing

program leading to the identification, selection,

and use of genetically superior boars, together

with guidelines for the development and use of

sow productivity and pig performance indexes. ”14 In the case of swine, the increased use

of intensive housing, which allows reproductive

control, should increase the impetus for progeny testing. Likewise, pinpointing areas where

considerable improvement remains to be made

should lead to the identification of selection

objectives.

Beef.—After World War II, a few breeders

became increasingly interested in problems of

inbreeding and the economic costs of dwarfism.

By that time, some had been trained in genetics

and some breed associations and State agencies

initiated localized testing programs for these

traits. In 1967, a “Beef Improvement Federation”

*There are several State programs—in Indiana, North Carolina,

and Tennessee. Some of these programs may test only growth and

not litter size.

14[1 .s. ~epartnlen~ of ~gri~ulture, ~gricultural Research Serv -

ire, AILS Na!ional Research Program, .Sn,ine Production, NRP No.

20370,” ()(’t()hw’ 1976.

of local and breed groups was formed to try to

consolidate the different systems of the State

improvement programs. The Federation is now

involved in: 15

. establishing uniform, accurate records,

. assisting member organizations in developing performance programs,

● Encouraging cooperation among all segments of the industry in using records,

● Encouraging education by emphasizing the

use of records,

● developing confidence in performance testing throughout the industry.

Despite these efforts, only about 3 percent of

beef cattle nationally are recorded. This relatively low participation rate, when compared

with NCDHIP, has both a technological and an

institutional explanation. Under the largely extensive beef raising system in the United States,

AI is difficult as long as estrus detection

technologies are unavailable. Natural stud service is usually more economical. Institutional barriers also prevent the development of a strong

genetic evaluation program–e.g., the breed

associations are not all eager to have their

breeds consistently compared with others. Likewise, some owners of bulls for stud service

would lose business in a strict testing scheme.

Goats .—Though little genetic work has been

done on goats in the past, the dairy goat industry has become more visible in the past few

years. The desire of goat breeders to participate

in NCDHIP led to the formation of a Coordinating Sub-Group for Dairy Goats. A review of

the research performed indicated a great need

for research in almost every area of production.

As a result, AIPL developed a plan for a genetic

improvement program. The leadership in the

dairy goat industry was convinced that it could

attain genetic improvement faster and at a

lower cost via NCDHIP than it could for any

other type of research.

In 1979, AIPL received a $15,000 grant from

the Small Farms Research Funding to supper

the development of genetic evaluation proce

ISR, [,. Willhanl, ‘i(;~neti(: AN k’itv in the 11. S. Beef [lld Ustl’.v,

JOUIYMI Paper No. J-7923 of the low;a Agricultur:tl itnd Home M

nomk!s I’:xperiment Station, Ames, l~wii, project No. 2?,()()(), n.cj.
Ch. 9—Advances in Reproductive Biology and Their Effects on Animal improvement . 183

dures for goats. Genetic evaluations for yield of

dairy goat bucks will be available before the end

of fiscal year 1980. Because limited genetic improvement for yield has occurred in dairy goats

in the past, these evaluations will probably have

a significant impact on the industry. AIPL can

virtually guarantee beneficial results because of

the data available from NCDHIP, its own expertise in genetics, statistics, and computer technology, and the decades of highly effective research on genetic improvement of dairy cattle

that can be adapted for the dairy goat industry.

However, funding for the goat testing program

remains on a year-to-year basis.

CONCLUSION

NCDHIP has shown how important genetic information is to the production of meat and dairy

products. The obstacles to such a program are

also formidable, but every failure to capitalize

on genetic potential is paid for by American

consumers. It has also shown that where selection objectives can be identified and agreed on,

and where conflicting interests can be brought

together to develop a program serving all interests, genetic improvement can become a central objective in breeding programs across the

country. Without reliable, evaluative data on

breeding stock, the Nation’s breeders will have

little interest in adopting new breeding technologies as they become available.

Impacts on breeding

An improvement in germplasm, like an increase in the nutritional content of fertilizer or

new and improved herbicides and pesticides, increases the quality of the physical capital used

on the farm. It is likely that much improvement

can still be made in the germplasm of all major

farm animal species using existing technology.

Selecting for desired characteristics causes a

specific qualitative change; it enhances the efficiency of the information contained within each

cell. The genetic information in each cell of a

farm animal is either more or less desirable or

efficient than information in the cells of another

animal, depending on how it performs on important traits. Superior germplasm can be used

in breeding decisions to upgrade a farmer’s

breeding or producing stock. (DHIA programs

are the best example of how information might

be distributed.)

Resources invested in genetics and in technologies related to genetics will have high payoffs—

e.g.) in a classic study16 of the payoff to research

in hybrid corn and in subsequent studies of

other types of genetic improvement, a high

cost/benefit ratio for such research was found.

The original study also showed that the absolute

market value of a particular product is an important factor influencing the rate of return on

a given research expenditure. In general, the

greater the aggregate value of the product, the

greater the rate of return on a research expenditure.17 Thus, the large expenditures for meat

and animal products in the United States suggest a great payoff in applied genetic research.

Beef purchases alone account for between 2 and

5 percent of the American consumer dollar, and

the total market value for beef is more than

twice that for corn in the United States.

DAIRY CATTLE

Total milk production has been stable for

many years. While milk production per cow has

gone steadily upward, the number of cows

during the past 35 years has decreased proportionately. (See figure 29.) Milk production per

cow should continue to increase, assuming that

no radical changes in present management systerns occur. The increase in production per cow

could continue even if no bulls superior to those

already available are found, simply as a result of

more farms switching to existing technology

and existing bulls. Moreover, bulls produced

from this system are increasing in superiority.

The number of dairy cows calved as of January 1, 1980, was 10,810)000. It has remained relatively stable for the past year, but may deIszvi ~l.iliches, “Research [;osts and Social Returns: H.vbrid {~01’1)

and Related Innovations, “Journal of Politica/ Economy 66:419, october 1958. See also R. E. Flrenson, P. E. Waggoner, and \’. W. Ruttan, “Economic Benefit From Research: An Example From Agriculture, ” Science 205: 1101, Sept. 14, 1979.

IW. Peterson and }’ujino Ha.vami, “Technical Change in Agriculture, ” Staff Papers series No. DP73-20, Depiirtment of Agrkxdture

and Applied klonomics, [ lnitwrsit. v of hlinmsota, St. Paul, Minn.,

July 1973.
184 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

crease to around 10 million in the next decade if

milk production continues to increase.

Artifical Insemination.—An example of

the interaction between technologies and genetic improvement is shown in table 34. The “predicted difference” (PD) in milk production represents the ability of individual bulls to genetically transmit yield—the amount of milk above

or below the genetic base that the daughters of

a bull will produce on average due to the genes

they receive. As indicated in table 34, the predicted difference for milk yield transferred via

the bull shows an improvement from 122 to 908

lb for active AI bulls in the United States over

the past 13 years.

This impressive improvement still lags behind

what is theoretically possible. A hypothetical

breeding program could result in an expected

yearly gain of 220 lb of milk per cow, using AI;

and the biological limits to this rate of gain are

not known. In practice, the observed genetic

trend in the U.S. national dairy herd is about

100 lb—70 lb from the PDs of bulls plus 30 lb or

so from the female, most of which is actually

carryover effect from the previous use of superior bulls.

AI organizations, many of which are cooperatively owned by dairymen, have not rigorously applied the principles of AI. Their efforts

have been limited by reluctance to break with

traditional selection practices, financial constraints for proper testing of young bulls to proTable 34.–Predicted Difference (PD)

of Milk Yield of Active Al Bulls

Year PD milk (lb)

1967. . . . . . . . . . . . . . . . . . . . . . . 122

1968. . . . . . . . . . . . . . . . . . . . . . . 198

1969. . . . . . . . . . . . . . . . . . . . . . . 205

1970. . . . . . . . . . . . . . . . . . . . . . . 276

1971 . . . . . . . . . . . . . . . . . . . . . . . 301

1972. . . . . . . . . . . . . . . . . . . . . . . 346

1973. ........, . . . . . . . . . . . . . 348

1974. ........, . . . . . . . . . . . . . 338

1975. . . . . . . . . . . . . . . . . . . . . . . 425

1976. . . . . . . . . . . . . . . . . . . . . . . 501

1977. . . . . . . . . . . . . . . . . . . . . . . 558

1978. . . . . . . . . . . . . . . . . . . . . . . 748

1979. . . . . . . . . . . . . . . . . . . . . . . 908

SOURCE: Animal Improvement Programs Laboratory, Animal Science Institute,

Beltsville Agricultural Research Center, USDA.

duce sires of cows, and too much emphasis on

nonproductive traits of questionable economic

value. The progress that has been made has resulted from the increased use of AI, the availability of data through NCDHIP, and the actual

use of reliable genetic evaluations. If any of

these three factors had been missing, far less

improvement would have occurred,

Semen Storage. –It is doubtful that major

technological changes in processing semen will

occur, However, since the rate of conception is

as important as the genetic merit of a sire to the

economy of a dairy enterprise, more attention

will be given to selecting sires of high fertility.

Progress should be made in banking semen by

AI studs as a hedge against costs of inflation. In

the future, some of the increased costs of housing and feeding bulls will probably be offset by

semen banking and earlier elimination of many

bulls.

Sexed Semen.—Sexing of semen to produce

heifer calves (for dairymen) or bull calves (for

AI organizations) has been attempted without

success for many years.

Perfect determination of the sex of progeny

could practically double selection intensity in

two ways—with dams to produce bulls for testing in AI and dams to produce replacements. If

sexed semen is used with an AI plan, the theoretical improvement in milk yield would be 33

lb per year, with 23 lb due to selection of dams

for replacements.

The value of this additional amount per year

may not seem great for any individual cow, but

when it is multiplied by a national herd of 7

million cows using AI and is accumulated for 10

years, the economic value, at $0.10/lb, is about

$1.1 billion—an average of$110 million per year

and $231 million during the l0th year. The cost

of sexing semen is not known, since no one has

successfully done it. If a way is found, the cost

would have to be under $10 per breeding unit

for the procedure to be economical.

Embryo Transfer.—The transfer of fertilized eggs from a cow to obtain progeny has

been accomplished with great success, Most

transfers have involved popular or exotic breed-
Ch. 9—Advances in Reproductive Biology and Their Effects on Animal improvement ● 185

ing animals with little regard for genetic potential.

Embryo transfer may never pay for itself in

terms of milk production of the animals produced except indirectly through bulls. Rather, it

is used mostly to produce outstanding cows for

sale. Other commercial applications for cattle

include obtaining progeny from otherwise infertile cows, exporting embryos instead of animals, and testing for recessive genetic traits,

Embryo transfer progeny must be worth

$2,500 each to justify the costs and risks. About

$1,500 of this represents costs due to embryo

transfer and $1,000 the costs of producing

calves normally. If genetic gain from embryo

transfer comes only from dam paths, the expected gain over AI alone is 76 lb/yr. Extra gain

at $0.05/lb above feed cost would have to accumulate for 79 years before added gain would

equal even a $300 embryo transfer cost per

pregnancy. If less semen is needed (allowing

more intensive bull selection), the expected gain

of 129 lb/yr must accumulate for 46 years to

balance an embryo transfer cost of $300 per

pregnancy.

Embryo transfer and perfect sexing of semen

would combine to improve genetic gain (in milk

production) slightly. The use of less semen

might be possible through application of in vitro

fertilization. However, feasibility based on

genetic gain would still require holding all costs

down to around $50 to $90 per conception. The

general conclusion is that costs of embryo transfer must be greatly reduced to be economically

feasible if only genetic gain is considered.

Estrus Synchronization.—The availability

of an effective estrus synchronization method

would provide strong impetus for increased use

of AI and embryo transfer in dairy cattle. The

detection of estrus is an expensive operation; effective control of estrus cycling also requires intensive management, adequate handling facilities, and close cooperation between the producer, veterinarian, and AI technician.

Summary.—

 Proper application of progeny testing with

selection and AI can increase the genetic

gain for milk yield more than two times

faster than is occurring today. Improved

evaluation of cows, proper economic emphasis on other traits, and strict adherence

to selection standards are the keys. Biological limitations to this rate of genetic improvement cannot be anticipated in the

foreseeable future.

AI of dairy cattle, with the present intensity of sire selection, should increase the net

worth or profit of animals (increased value

minus extra costs of the AI program) about

$10.00/head per year. By 1990, 8 million

dairy cows in AI programs would be worth

about $800 million (8 X 106 X 10 X 10

years) more at current market prices as a

result of continued use of AI.

Sexing of semen when used with AI may

pay for itself if the cost per breeding unit

can be kept between $10 and $20.

Embryo transfer is unlikely to pay for itself

genetically unless the cost is reduced to between $50 and $90 per conception. However, despite its high costs, it is used to produce animals of exceptionally high value.

(See app. II-C for an explanation of reasons

other than genetics why embryo transfer is

used. )

Estrus synchronization is now available for

use with heifers, and should increase the

use of AI and consequently the genetic improvement of dairy cattle.

A secondary benefit of all technologies is

the increased number of skilled persons

who can provide technical skills as well as

educate dairymen in all areas. Also, a

unique pool of reproductive and genetic

data has been accumulated.

BEEF CATTLE

There is no single trait of overriding ire-’

portance (like milk production in dairy cows) to

emphasize in the genetic improvement of beef

cattle, the rate of growth is a possibility. * It is

also difficult to select for several traits at once,

● Beef and dairy cattle are usually different breeds in the LJnited

States. In the literature and in research they are often referred to

as dit”ferent species. In other countries, notably in Western Europe

and in Japan, so-called “dual purpose” cattle are used to produce

both beef and milk. In the [Jnited States, old dairy cows usually become hamburger.
186 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

especially when some are incompatible—e.g., it

is desirable to produce large animals to sell, but

undesirable to have to feed large mothers to

produce them. There are also other complications. Growth rate has two genetic components,

for which one can select—the maternal contribution (primarily milk production) and the

calf’s own growth potential. Other traits of interest are efficiency of growth, carcass quality

traits (such as tenderness), calving ease, and

reproductive traits, such as conception rate to

first service with AI,

Genetic improvement programs for beef have

two major advantages over those for dairy cattle traits such as growth rate and carcass quality

can be measured in both sexes (whereas one

cannot measure the milk production of bulls);

and the traits are more heritable than milk

production.

Artificial insemination.—Between 3 a nd

5 percent of the U.S. beef herd is artificially inseminated each year. This low rate is due to several factors, including management techniques

(range v. confined housing), availability of related technologies (especially, until recently,

estrus synchronization), and the conflicting objectives of the individual breeders, ranchers,

and breed associations.

Because little is known about the effectiveness of Al in spreading specific genes throughout the Nation’s beef herds, analysts have concentrated on their reproductive performance.

Calf losses are heavy throughout the Nation.

The calf crop-the number of calves alive at

weaning as a fraction of total number of females

exposed to breeding each year—is estimated to

be between 65 and 81 percent. To put these

data in perspective, USDA18 has estimated that a

5-percent increase in the national calf crop

would yield a savings of $558 million per year in

the supply of U.S.-grown beef. Techniques now

available can produce such an increase when

they are integrated into an adequate management program.

The standardized measure of weaning weight

in beef cattle is the weight at 205 days, adjusted

for sex of calf and age of dam. In a recent study

in West Virginia—the Allegheny Highlands Project—calf weights have averaged an increase of

10 lb per year of participation in the project, via

AI and crossbreeding. Estimates of increased

value of calves statewide, should the same tests

and AI program be expanded, add up to $3.6

million per year when calf prices average $50

per hundredweight.l9 Rapid adoption of AI

could bring about this kind of increase in as little as 40 to 48 months.

The costs and returns of AI vary from farm to

farm and with the number of cattle in estrus. In

general, it becomes more valuable with smaller

herds, more cows in estrus, higher conception

rates, and better bulls. For purebred herds,

even larger benefits have been estimated—e.g.,

in a 1969 study, the estimated increase in value

per calf when AI was used was $30.02 on purebred ranches compared to $3.31 on commercial

ranches in Wyoming.20

A major secondary, or indirect, benefit of the

use of AI is feed saved for other uses. It has

greatly reduced the number of sires necessary

for stud service and, through radically improved milk production, the number of females

as well. These reduced requirements together

are equivalent to more than 1 billion bu of corn

and other concentrates. This situation will be

further enhanced as beef cattle AI expands.

synchronization of Estrus.—Differences

in the rates of application of AI between beef

and dairy herds can be explained partly by the

differing management systems for the two

types of classes of cattle. Dairy herds are kept

close to the barn for milking and are accustomed to being approached by humans. In contrast, beef herds may number a few thousand

head on 100,000 acres of arid pasture land. The

detection of estrus under these conditions is

difficult.

IM[ I s, [)t,,,i,,.l ,,,(,,11 of ~~gl.i[.llltlll.p, t\gricuttural Research Sel~’-

i(.v, “Beet Production,” /\RS Niitio!lil] Research Pi’Og]siIM Repel’t No.

20360” (\ Vilshillgtoll, [).(;.: ( ISI)/\, ()(-t(}hel’ 19761.

MB. s, ~iik~l., AI. R. ~’~llsett, P. k:. Lewis, and F.. K. lnskq) “A

P,.(,gl.:llll RePolq 011 the Allegheny Highlands Project” Uvlorgantown, W. \’a.: West t’irginia University, January-December 19791.

Z(j[). ~l. st[?l,ells and “I’. M~hl’, “Artificial Insemination of Range

(httle in \V~onling: An Economic Analysis, ” Wyoming Agricultural

k;xperiment Stiitk)n Bulletin No. 496, 1969.
Ch. 9—Advances in Reproductive Biology and Their Effects on Animal Improvement ● 187

It has been predicted that the availability of

prostaglindin agents for regulating estrus could

increase the number of beef calves born from

superior bulls by 10 times, and that perhaps 20

percent of the U.S. beef cow herd could receive

at least one insemination artificially by 1990.2

1 If

this lead to a 50-lb increase in weight for 10 percent of the calves born, it should be worth $114

million to $122 million each year, assuming 80

or 85 percent net calf crop and $60 per hundredweight.

The implementation of recently developed

estrus synchronization technology might increase the number of beef cows bred artificially

by 4,000,000 in the United States. Such a program should be successful in advancing the

calving date by one week (by decreasing the

calving interval), and in increasing the quality of

the calves produced. These new calves could be

worth about $100 million annually, less about

$50 million due to extra costs associated with

the synchronization program.

Sex Control.—Sex control would have a

dramatic effect on the beef industry. In 1971, it

was projected that by 1980 sex control could

have an annual potential benefit of $200 million based on 10 million female calves being replaced by male calves produced through the

sexing of semen. 22 At the time of the prediction,

the market value for steers was about $20 more

than for heifers. (Steers wean heavier and gain

more efficiently. ) Now the margin is much

greater–approximately $50. This potential

method of biological control is more attractive

than the use of additives like steroids or implants because of the possible hazards associated with them that preclude their use.

Embryo Transfer.—The possibilities for

genetic improvement in beef cattle using embryo transfer have been analyzed. It appears

that embryo transfer programs can be developed to increase the rate of genetic progress for

ZIH. c). Hafs, “potential Impact of Prostaglandin on prospects for

F’ood f’rom Dairy Cattle,” Proc. L.ufa/vse Symposium, J. w. Lauderdak and J. H. Sokolowski (eds. ) (Kalamazoo, Mich.: Lfpjohn, 19791,

pp. 9-14.

UK. H. F’oot[? ~IMl P. Nli]ler, “what Might Sex Ratio Contro] Mean

in the Animal World,’” S.vmposium, Am. Soc. of Animal Science,

1971, pp. 1-1o.

growth rate; but the programs are much too expensive to be used over the entire population.

One problem is that the economic value of the

product of a beef cow is around 25 percent (or

even less) of that of a dairy cow. Nevertheless,

in populations in which AI is used, embryo

transfer was found to be useful for obtaining

more bulls from top cows. The females produced by embryo transfer would be worth marginally more than females produced conventionally, but the costs and influence of males

could spread over the population through the

use of AI. The extent of this use of embryo

transfer would be very small; only a few hundred bulls would be produced per year for very

large populations, and over 99 percent of the

population would reproduce conventionally.

However, such programs could have considerable economic benefit. Care must be taken to

minimize increased inbreeding of the population with such a breeding scheme.

Summary.—

●

●

●

AI could substantially improve economically important traits in beef herds. However,

because of the diversity of traits considered important by different breed groups

and the lack of a national beef testing and

recording system comparable to NCDHIP,

economic estimates of its value have not

been developed.

A sexing technology to produce mostly

males (they grow faster than heifers) could

be of enormous potential benefit to the

beef industry. However, no successful

technique yet exists.

Estrus cycle regulation could lead to a substantial increase in the number of beef cattle in AI programs. The net benefit of this

technology, coupled with AI, may be as

high as $50 million per year. Similarly, the

availability of reliable progeny records

would add to the beneficial impact of AI in

beef and would probably contribute significantly to its use in beef cattle.

OTHER SPECIES

Swine.—Much progress has been made in

improving the overall biological efficiency of

pork production in the United States. Improved
188 ● Impacts of Applied Genetics —Micro-Organisms, Plants, and Animals

growth rates, feed efficiencies, carcass merit,

and litter sizes have helped keep pork prices

down and improve its quality in the Nation’s

markets. Pork today is leaner and contains more

high-quality protein calories than it was just a

few decades ago.

AI in swine production could expand, although it will be limited by the relatively poor

ability of swine sperm to withstand freezing

and by the problem of detecting estrus. It will

be encouraged by the strong trend toward confinement housing and integration of all phases

of hog production. The industry—especially the

individual, family-farm type units—would benefit by the establishment of a progeny testing

scheme to identify superior boars. Publicly

available information on genetic merit would

decrease dependence on a few corporate breeding organizations.

Embryo transfer in swine will be strictly

limited by difficulties in developing nonsurgical

methods of recovery and transfer, and by the

low economic value per animal in comparison to

cattle and horses. However, embryo transfer is

useful in introducing new genetic material into

breeding herds of specific pathogen-free swine

and in transporting genetic material to various

regions of the world.

Sheep. —The processes of selection and of

crossing specific strains, which have been so effective in poultry and hogs, have been virtually

ignored in sheep. Selection of replacement ewes

from the fastest growing ewe lambs born as

twins and the use of flushing to increase ovulation rates have led to annual increases of 1.8

percent in lambing; in one test the market

weight of lambs ws increased by one lb per year

of cooperation .23

Synchronization of estrus in ewes can be

achieved with prostaglandin and many different progestogens. The technique is used extensively in many countries, but no products for

this purpose are currently marketed in the

United States.

AI rates abroad sometimes approach 100 percent. However, AI will not be used widely on

sheep in the United States until systems for performance and progeny testing are implemented

that will track the number of lambs born and

their growth rate, and until routine freezing of

raw semen is achieved.24

Goats .—The research performed on goats is

largely designed for application to other animals. However, interest in goats in the United

States and the demand for their products

through the world is increasing.

NCDHIP has just started providing sire evaluations to goat breeders. These data, along with

artifical insemination, should increase milk production. The genetic data might be of particular

usefulness in the less developed countries

where most goat raising occurs. Greater use of

all reproductive technologies on valuable Angora goats might be expected.

Other technologies

The use of any reliable twinning or sex selection technologies will be limited until such procedures can be made simple, fast, inexpensive,

and innocuous. No widespread use of these

technologies should be expected within the next

decade.

The more esoteric techniques for manipulating sex cells or the germplasm itself will have

no impact on the production of animals or

animal products within the next 20 years. In

vitro manipulations, including cloning, cell fusion, the production of chimeras, and the use of

rDNA techniques, will continue to be of intense

interest. However, it is unlikely that they will

have practical effects on farm production in the

United States in this century. Each technique

will require more research and refinement. Until specific genes can be identified and located,

no direct gene manipulation will be practicable.

A polygenic basis for most traits of importance

can be expected to be the rule rather than the

exception.

Should such techniques become available,

limited use for producing breeding stock can be

expected. Experience with early users of AI and

z3K;. K. Illskeep, personal Lmllllllllllicatioll, 1980. Z41bid.
Ch. 9—Advances in Reproductive Biology and Their Effects on Animal improvement ● 189

embryo transfer is strong evidence for the predicted use of the technologies, no matter what

their economic justification. (See app. II-C.)

A major, secondary effect of animal research

in reproductive biology is increased understanding leading to the possible solution of

human problems—e.g., the concept, efficacy,

and safety of the original contraceptive pill was

developed and established in animals. It involves the same principle as estrous cycle regulation discussed above.

AQUACULTURE

Aquiculture is the cultivation of freshwater

and marine species (the latter is often referred

to as mariculture). While fish culture is about

6,000 years old, scientific understanding of its

basic principles is far behind that of agriculture.

Aquiculture is slowly being transformed into a

modern multidisciplinary technology, especially

in the industrialized countries. Increasing

awareness of human nutritional needs, overfishing of natural commercial fisheries, and rising worldwide demand for fish and fish products are trends that indicate a growth in interest in aquiculture as a means to meet the food

needs of the world’s population.

As part of the trend toward the high technology and dense culturing of intensive aquaculture systems in the industrialized countries,

problems of reproductive control, hatchery

technology, feeds technology, disease control,

and systems engineering are all being investigated. Reproductive control and genetic selection are important because most commercial

aquiculture operations must now depend on

wild seedstocks. Very little information on the

animals in culture is available.

With all three of the aquiculture genera (fish,

mollusks, and crustaceans), selective breeding

programs have long been established, healthy

gene pools are available, and advantageous hybridizations have been developed. In fish raising, culture systems often demand sterile hybrids, especially of carp and tilapia, Selective

breeding of salmon has been limited by political

pressures. Very little work has been conducted

with catfish, the largest aquiculture industry in

the United States. The use of frozen sperm,

which has been successful, should increase because of the savings in transport costs. Although

culture systems for mollusks are fairly welldefined, little applied genetics work has been

done with these popular marine species. Some

success has been reported in selection for

growth rate and disease resistance of the

American oyster, and selection for growth rate

of the slow-growing abalone is underway. The

crustaceans, of which the Louisiana crayfish is

the largest and most viable industry, are the

least understood. Successful hybrids of lobsters

have been developed.

Aquaculture suffers from an insufficient research base on the species of interest. However,

growing appreciation of and demand for marine species should result in increased support

for basic and developmental work on all aspects

of control, including basic reproductive biology.

POULTRY BREEDING

The quantitative breeding practices of commercial breeders have changed very little over

the last 30 years. Highly heritable traits, such

as growth rate, body conformation, and egg

weight, are perpetuated by mass selection because little advantage is gained from hybrid

vigor. Low heritable traits (egg production, fertility, and disease resistance) are perpetuated by

crossbreeding and identified through progeny

and family testing.

The goals of the industry are to increase egg

production of the layers–both in quality and

quantity–and, with broilers and turkeys, to improve growth rate, feed efficiency, and yield, as

well as to reduce body fat and the incidence of

defects.

The technologies of AI and semen preservation have accelerated the advances made

through quantitative breeding technology. AI is

widely used in commercial turkey breeding because of the inability of modern strains to mate.

It makes breeding tests more efficient, steps up

selection pressure on the male line, reduces the

number of necessary breeder males, and increases the number of females that may be

mated to one male. Semen diluents were introduced to the turkey industry about 10 years ago

to lower the cost of AI. Currently, a little over
190 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

half of the turkeys are inseminated with diluted

semen.

Preservation of poultry semen by freezing is

now practiced by several primary breeders. Although freezing chicken semen causes it to lose

some potency, the practice allows increased genetic advancement and the distribution of genetic material worldwide.

The amount of genetic variation available for

breeding stock is not expected to diminish in the

near future. Ceilings for certain traits will eventually be reached, but certainly not in the

1980’s. Advances in breeding laying chickens

will be less dramatic than in the past, but efforts

will continue to develop new genetic lines and

to improve reserve lines and crosses to meet future needs.

The growth rate of broilers will continue t.

increase at 4 percent a year, which suggests

that birds will be reaching 4.4 lb in 5 weeks by

the 1990’s. Breeding for stress resistance will be

increasingly important, not only because of the

increased use of intensive production systems,

but also to meet the physiological stresses resulting from faster growth and greater weight.

AI will assume increasing importance. Recent

advances in procedures for long-term freezing

of chicken semen will allow breeders to extend

the use of outstanding sires. The sale of frozen

seman may eventually substitute, in part, for

the sale of breeder males.

Dwarf broiler breeders will also assume increasing importance over the new few years.

The dwarf breeder female is approximately 25-

percent smaller than the standard female, and

even though the dwarf’s egg is smaller and the

progeny’s growth rate slightly less than that of

the standard broiler, the lower cost of producing broiler chicks from the dwarf breeder more

than offsets the slight loss in their growth rate.

Dwarf layers and the dwarf breeder hens could

reduce production costs by 20 percent and 2

percent, respectively.

There is some interest among poultry breeders in cloning, gene transfer, and sex control

but progress toward successful technologies is

slow.

Issue and Options for Agriculture—Animals

ISSUE: Should the United States increase support for programs in

applied genetics for animals and

animal products?

Advocates of a strong governmental role in

support of agricultural research and development (R&D) have traditionally referred to the

small size of the production unit: U.S. farms are

too small to support R&D activities. Throughout

this century a complicated and extensive network of Federal, State, and local agricultural

support agencies has been developed to assist

the farmer in applying the new knowledge produced by research institutions. This private/

public sector cooperative network has produced an abundant supply of food and fiber,

sometimes in excess of domestic demand. Socially oriented policies have been adopted to soften

the impacts of new technology and to rescue the

marginally efficient farmer from bankruptcy.

Current projections of U.S. and world population growth show increasing demand for all

food products. Other predictable trends with

implications for agricultural R&D, include:

●

●

●

●

●

growth in income for some populations,

which will probably increase the demand

for sources of meat protein;

increasing competition among various

sources of protein for the consumer’s

dollar;

increasing awareness of nutrition issues

among U.S. consumers;

increasing competition for prime agricultural land among agricultural, urban, and

industrial interests;

increasing demand for U.S. food and fiber
Ch. 9—Advances in Reproductive Biology and Their Effects on Animal improvement ● 191

products from abroad, leading to opportunities for increased profits for successful

producers; and

. increasing demands on agricultural products for production of energy.

OPTIONS:

A. Governmental participation in, and funding of,

programs like the National Cooperative Dairy

Herd Improvement Program (NCDHIP) could

be increased. The efforts of the Beef Cattle

Improvement Federation to standardize procedures could be actively supported, and a

similar information system for swine could be

established.

The fastest, least expensive way to upgrade

breeding stock in the United States is through

effective use of information. Computer technology, along with a network of local representatives for data collecting, can provide the individual farmer or breeder with accurate information on the germplasm available, so that he

can then make his own breeding decisions. In

this way, the Nation can take advantage of population genetics and information handling capabilities to upgrade one of its most important

forms of capital: poultry and livestock. Breed

associations and large ranchers who sell the

semen from their prize bulls based on pedigrees

rather than on genetic merit may act as barriers

to the effectiveness of such an objective information system.

The benefits of such programs would accrue

both to U.S. consumers, in reduced real prices

of meat and animal products, and to producers

who participate in the programs, in increased

efficiency of production. Consumers spend such

a large part of their incomes on red meat that

every increase in efficiency represents millions

of dollars saved. Beef producers too, should

welcome any assistance in upgrading their

stocks. The price of semen has remained relatively stable, and semen from bulls rated

highly on certain economic traits costs only a

few dollars more than that from average bulls.

However, efficiency of production is not the

only value to be upheld in U.S. agriculture—e.g.,

in milk production complex policies have been

designed to maintain constant milk supplies

without large fluctuations in price.

The NCDHIP model program for dairy cattle

has shown that an effective national program

requires the participation by the varied interests in program policymaking in an extension

network, for local collection and validation of

data and for education and of expertise in data

handling and analysis. Also important is a

strong leadership role in establishing the program. This option implies that the Federal Government would play such a role in new programs and expand its role in existing ones.

B. Federal funding of basic research in total animal improvement could be increased.

The option, in contrast with option A,

assumes that it is necessary to maintain or expand basic R&D to generate new knowledge

that can be applied to the production of improved animals and animal products.

Information presented in this report supports

the conclusion that long-term basic research on

the physiological and biochemical events in

animal development results in increasing the efficiency of animal production, both in total

animal numbers and in quality of product. Increased understanding of the interrelationships

among various systems—including reproduction, nutrition, and genetics—gradually leads to

the development of superior animals that efficiently consume food not palatable to humans

and are resistant to disease.

Earlier studies also support the importance of

basic research–e.g., the National Research

Council found in 1977 that”. , . not as much fundamental research on animal problems has

been conducted in recent years . . . it should

receive increased funding. ”25 USDA also found,

in a review of various conference proceedings,

congressional hearings, special studies, and

other published materials on agricultural R&D

priorities, strong support for more research on

the basic processes that contribute to reproduction and performance traits in farm animals:

2sN~~t iollill [~f,$(,ill.(,ll (hllll(;il, Worki FIIod aIId Nutrition .stlJd.Vt

‘/’/If; Polfmlia/ (,’onlrilmlions qf’ Rfksfwrch ILVilShill#Oll, l). (:. illlttlol’,

1977), p. 97.
192 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Specific livestock research areas identified as

having significant potential for increased production both in the United States and developing countries include: 1) control of reproductive

and respiratory diseases, 2) developing genetically superior animals, 3) improving nutrition

efficiency, and 4) increasing the reproductive

performance of all farm animal species.26

z611. S. Department of A#ictdture, Science and Education Administration, Agricuhura/ and Food Research Issues and Priorities

(Washington, D. C.: author, 1978), p. xiii.

Regardless of the effectiveness of present

population control programs or of current

trends in individual decisions about family size,

the output of the Nation’s agricultural activities

must increase over the next decades if sufficient

food is to be available for the world’s population. Basic research is the source from which

new applications to increase productivity arise.
Part III

Institutions and Society

Chapter 10. The Question of Risk . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 197

Chapter 11. Regulation of Genetic Engineering. . . . . . . . . . . . . . . . . , . . . . . . . . . . . . . .211

Chapter 12. Patenting Living Organisms. . . . . . . . . . . . . . . . . . . . . . . , . . . . . . . . . . . . .237

Chapter 13. Genetics and Society . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .257
Chapter 10

The Question of Risk
Chapter 10

Page

Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . 197

The Initial Fear of Harm . . . . . . . . . . . . . . . . . . . 197

Classification of Potential Harm . . . . . . . . . . . . 198

Identification of Possible Harm . . . . . . ., . . . . zoo

Estimates of Harm: Risk. . ................200

The Status of the Current Assessment of

Physical Risk . . . . . . . . . . . . . . . . . . . . . . . 201

Perception of Risk . . . . . . . . . . . . . . . . . . . . . .203

Burden of Proof . . . . . . . . . . . . . . . . . . . . .. 203

Other Concerns . . . . . . . . . . . . . . . . .........204

Concerns Raised by Industrial Applications .. .204

Concerns Raised by the Implications of the

Recombinant DNA Controversy for General

Microbiology. . . . . . . . . . . . . . . . . . . . . .. ..204

Concerns Raised by the Implications of the

Recombinant DNA for Other Genetic

Manipulation . . . . . . . . . . . . . . . . . . . . .. ..206

Page

Ethical and Moral Concerns. , . . . . . . . . . . . . .207

Conclusion . . . . . . . . . . . . . . . . . . . . .........207

Figures

Figure No. Page

35. Flow Chart of Possible Consequences of Using

Genetically Engineered Micro-Organisms , . . 199

36. Flow Chart to Establish Probability of Harm

Caused by the Escape of a Micro-Organism

Carrying Recombinant DNA . .............201

37. Alternative Methods for Transferring DNA From

One Cell to Another. . . . . . . . . . . . . . . . . . . . 206
Chapter 10

The Question of Risk

Introduction

The perception that the genetic manipulation

of micro-organisms might give rise to unforeseen risks is not new. The originators of chemical mutagenesis in the 1940’s were warned that

harmful uncontrolled mutations might be induced by their techniques. In a letter to the

Recombinant DNA Advisory Committee (RAC) of

the National Institutes of Health (NIH) in December of 1979, a pioneer in genetic transformation

at the Rockefeller University, wrote: “. . . I did

in 1950, after some deliberation, perform the

first drug resistance DNA transformations, and

in 1964 and 1965 took part in early warnings

against indiscriminate ‘transformations’ that

were then being imagined. ”1

Yet none of this earlier public concern led to

as great a controversy as has research with recombinant DNA (rDNA). No doubt it was encouraged because scientists themselves raised

questions of potential hazard. The subsequent

open debates among the scientists strengthened

the public’s perception that there was legitimate

cause for concern, This has led to a continuing

attempt to define the potential hazards and the

chances that they might occur.

‘ Rollin 1). Ho[chkiss, Recombinant DNA Research, vol. 5, NIH publication No. 80-2130, March 1980, p. 484.

The initial fear of harm

For the purposes of this discussion, harm (or

injury) is defined as any undesirable consequence of an act. Such a broad definition is warranted by the broad targets for hypothetical

harm that genetic manipulation presents: injury

to an individual’s health, to animals, to the environment.

The inital concern involved injury to human

health. Specifically, it was feared that combining the DNA of simian virus 40, or SV40, with an

Escherichia coli plasmid would establish a new

route for the dissemination of the virus. Although the SV40 is harmless to the monkeys

from which it is obtained, it can cause cancer

when injected into mice and hamsters. And

while it has not been shown to cause cancer in

humans, it does cause human cells to behave

like cancer cells when they are grown in tissue

culture. What effect such viruses might have if

they were inserted into E. coli, a normal inhabitant of the human intestine, was unknown.

This uncertainty, combined with an intuitive

judgment, led to a concern that something

might go wrong. The dangerous scenario went

as follows:

● SV40 causes cells in tissue culture to behave like cancer cells,

● SV40-carrying E. coli might be injected accidently into humans,

● humans would be exposed to SV40 in their

intestines, and

● an epidemic of cancer would result.

This chain of connections, while loose, was

strong enough to raise questions in at least some

people’s minds.

The virus SV40 has never actually been

shown to cause cancer in humans; but the potential hazards led the Committee on Recombinant DNA Molecules of the National Academy of

Sciences (NAS) to call in 1974 for a deferment of

any experiments that attempted to join the DNA

of a cancer-causing or other animal virus to vector DNA. At the same time, other experiments,

197

76-565 0 - 81 - 14
198 ● Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

that were thought to have a potential for harm

–particularly those that were designed to

transfer genes for potent toxins or for resistance to antibiotics into bacteria of a different

species—were also deferred. Finally, one other

type of experiment, in which genes from higher

organisms might have been combined with vectors, was to be postponed. The fear was that latent “cancer-causing genes” might be inadvertently passed on to E. coli.

Throughout the moratorium, one point was

certain: no evidence existed to show that harm

would come from these experiments. But it was

a possibility, The scientists who originally raised

questions wrote in 1975: “. . . few, if any, believe

that this methodology is free from risk. ”2 It was

recognized at that time that “. . . estimating the

risks will be difficult and intuitive at first but

this will improve as we acquire additional

knowledge. ”3 Hence two principles were to be

followed: containment of the micro-organisms

(see table 35, p. 213) was to be an essential part

of any experiment; and the level of containment

was to match the estimated risk. These principles were incorporated into the Guidelines for

Research Involving Recombinant DNA Molecules, promulgated by NIH in 1976.

But the original fears surrounding rDNA research progressed beyond concern that humans

might be harmed. Ecological harm to plants, animals, and the inanimate world were also considered. And other critics noted the possibility of

moral and ethical harm, which might disrupt

both society’s structure and its system of values.

Classification of potential “

physical harm

Some combinations of DNA may be harmful

to man or his environment—e.g., if an entire

DNA copy of the poliovirus genetic material is

combined with E. coli plasmid DNA, few would

argue against the need for careful handling of

this material.

For practical purposes, the potential harm

associated with various micro-organisms is

Viecombimm[ DNA Research, vol. 1, DHEW publication No. (NIH)

76-1138, August 1976, p. 59.

31bid.

shown in figure 35. Each letter (A through L)

represents the consequence of a particular combination of events and micro-organisms. For example, the letters:

A)C

B,D

E, I

F)J

H,L

G)K

represent the intentional release of microorganisms known to be harmful to the

environment or to man—e.g., in biological warfare or terrorism.

represent the inadvertent release of

micro-organisms known to be harmful to

the environment or to man—e.g., in accidents at high-containment facilities

where work is being carried out with

dangerous micro-organisms.

represent the intentional release of microorganisms thought to be safe but which

prove harmful—when the safety of organisms has been misjudged.

represent the intentional release of microorganisms which prove safe as expected—

e.g., in oil recovery, mining, agriculture,

and pollution control.

represent the inadvertent release of

micro-organisms which have no harmful

consequences— e.g., in ordinary accidents

with harmless micro-organisms.

represent the inadvertent release of

micro-organisms thought to be safe but

which prove harmful—the most unlikely

possible consequence, because both an

accident must occur and a misjudgment

about the safety must have been made.

Discussions of physical harm have recognized

the possibility of intentional misuse but have

minimized its likelihood. The Convention on the

Prohibition of the Development, Production,

and Stockpiling of Bacteriological (Biological)

and Toxin Weapons and on their Destruction

which was ratified by both the Senate and the

President in 1975, * states that the signatories

will “never develop . . . biological agents or toxins . . . that have no justification for prophylactic, protective, or other peaceful purposes. ”

Such a provision clearly includes micro-organisms carrying rDNA molecules or the toxins

4[;onvention of the Prohibition of the Development, Production,

and Stockpiling of Bacleriolo~ical (Biological) and ‘1’oxin Weapons

and on Their Destruction, Washin@on, Umdon, and Moscow,

Apr. 10, 1972; entered into force on Mar. 26, 1975 (26 [J. S.”r., 583).

*As of 1980, 80 countries hate ratified the treaty; another 40

have signed but not ratified.
Ch. 10—The Question of Risk . 199

Figure 35.—Flow Chart of Possible Consequences of Using Genetically Engineered Micro-Organisms

SOURCE: Office of Technology Assessment.

produced by them. It must be assumed that

those who signed did so in good faith.

While there is no way to judge the likelihood

of developments in this area, the problems that

would accompany any attempt to use pathogenic micro-organisms in warfare—difficulties in

controlling spread, protection of one’s own

troops and population—tend to discourage the

use of genetic engineering for this purpose. *

Similarly, the danger that these techniques

might be used by terrorists is lessened by the

scientific sophistication needed to construct a

more virulent organism than those that can

* A 11 hou~h storkpil in~ of” biological ma rf’are agents is prohibited,

l’(’S(’ill’(’tl into 11(W’ il#lllS is Ilot.

1

already be obtained—e.g., encephalitis viruses

or toxin-producing bacteria like C. botulinum or

C. tetani.

Some discussions have centered around the

possibility of accidents caused by a break in containment. Construction of potentially harmful

micro-organisms will probably continue to be

prohibited by the Guidelines; exceptions will be

made only under the most extraordinary circumstances. To date, no organism known to be

more harmful than the organism serving as the

source of DNA has been constructed.

However, the biggest controversy has centered around unforeseen harm—that microorganisms thought safe might prove harmful.
200 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Discussion of this kind of harm is hindered by

the difficulty not only of quantifying the probability of an occurrence but also of predicting

the type of damage that might occur. The different types of damage that can be conjured up are

limited only by imagination. The scenarios have

included epidemics of cancer, the spread of oileating bacteria, the uncontrolled proliferation

of new plant life; and infection with hormoneproducing bacteria.

The risk of harm refers to the chance of harm

actually occurring. In the present controversy,

it has been difficult to distinguish the possible

from the probable. It is, for instance, possible

that an individual will be killed by a meteor falling to the ground, but it is not probable. Analogous situations exist in genetic engineering. It is

in this analysis that debate over genetic engineering has some special elements: the uncertainty of what kind of harm could occur, the uncertainty about the magnitude of risk, and the

problem of the perception of risk.

Identification of possible harm

The first step in estimating risk is identifying

the potential harm. It is not very meaningful to

ask: How much risk does rDNA pose? The concept of risk takes on meaning only when harm is

identified. The question should be: What is the

likelihood that rDNA will cause a specific disease such as in a single individual or in an entire

population? The magnitude of the possible harm

is incorporated in the question of risk, but differs in the two cases. A statement about the risk

of death to one person is different than one

about the risk of death to a thousand. The right

questions must be asked about a specific harm.

Since no dangerous accidents are known to

have occurred, their types remain conjectural.

Identifying potential harm rests on intuition and

arguments based on analogy. Even a so-called

risk experiment is an approximation of subsequent genetic manipulations. That is why experts disagree. No incontestable “scientific

method” dictates which analogy is useful or acceptable. By their very nature, all analogies

share some characteristics with the event under

consideration but differ in others. The goal is to

discover the one that is most similar and to

observe it often. This process then forms the

basis for extrapolation.

For example, it has been argued that ecological damage can be caused by the introduction of

plants, animals, and micro-organisms into new

environments. Scores of examples from history

support this conclusion. The introduction to the

United States of the Brazilian water hyacinth in

the late 19th century has led to an infestation of

the Southern waterways. Uncontrolled spread

of English sparrows originally imported to control insects has made eradication programs necessary. Countless other examples are confirmation that biological organisms may, at times,

cause ecological damage when introduced into a

new environment. Yet there is no agreement on

whether such analogies are particularly relevant to assessing potential dangers from genetically engineered organisms. It could be argued-e.g., that a genetically engineered organism (carrying less than 1 percent new genes) is

still over 99 percent the same as the original,

and is therefore not analogous to the “totally

new” organism introduced into an ecosystem.

Some experts emphasize the differences between the situations; others emphasize the similarities.

Other analogies have been raised. New

strains of influenza virus arise regularly. Some

can cause epidemics because the population,

never before exposed to them, carries no protective antibodies. Yet can this analogy suggest

that relatively harmless strains of E. coli might

be transformed into epidemic pathogens? There

is disagreement, and debates continue about

what “could happen” or what is even logically

possible.

Estimates of harm: risk

Assuming that agreement has been reached

on the possibility of a specific harm, what can be

done to ascertain the probability? What is the

likelihood that damage will occur?

Damage invariably occurs as the result of a

series of events, each of which has its own particular chance of occurring. Flow charts have

been prepared to identify these steps. A typical
Ch. 10—The Question of Risk . 201

analysis determines a probability value for each

step-e. g., in figure 36 step II the probability of

escape can be estimated based on the historical

record of experiments with micro- organisms.

Depending on the degree of containment, the

probability varies. It is almost certain that

experiments on an open bench top, using no

precautions, will result in some escape to the

surrounding environment—a much less likely

event in maximum containment facilities. (See

table 35.)

Two points should be noted. First, each probability can be minimized by appropriate control

measures. Second, the probability that the final

event will occur is equal to or less likely than the

least likely link in the chain. Because the probabilities must be multiplied together, if the

probability of any single step is zero, the probability of the final outcome is zero; the chain of

events is broken.

THE STATUS OF THE CURRENT ASSESSMENT

OF PHYSICAL RISK

A successful risk assessment should provide

information about the likelihood and magnitude

of damage that might occur under given circumstances. It is clear that the more types of

damage that are identified, the more risk assessments must be carried out.

Figure 36.— Flow Chart to Establish Probability of

Harm Caused by the Escape of a Micro-Organism

Carrying Recombinant DNA

Event Probability

L Inadvertent incorporation of hazardous gene PI

into micro-organism

11. Escape of micro-organism into environment P,

Ill. Multiplication of micro-organism and P3

establishment in ecological niche

V. Production of factor to cause disease

P*

NOTE: P, will always be smaller than any of the other probabilities.

SOURCE: Office of Technology Assessment.

Although the original charter of RAC underscored the importance of a risk assessment program, it was not until 1979 that the details of a

formal program were published. For 5 years,

risks were assessed on a case-by-case basis

through: 1) experiments carried out under contract from NIH, 2) experiments that were designed for other purposes but which proved to

be relevant to the question of risk, and 3) conferences at which findings were examined.

From the start, it was difficult to design experiments that could supply meaningful information-e.g., how does one test the possibility

that “massive ecological disruptions might

occur?” Or that a new bacterium with harmful

unforseen characteristics will emerge? Still

some experiments were proposed. But because

these experiments had to be approximations of

the actual situation, the applicability of their

findings was debated. Here too, experts could

and did disagree—not about the findings themselves, but about their interpretation.

For example, in an important experiment designed to test a “worst case situation, ” a tumor

virus called polyoma was found to cause no

tumors in test animals when incorporated into

E. coli.5* Since just a few molecules of the viral

DNA are known to cause tumors when injected

directly into animals, it was concluded that

tumor viruses are noninfectious to animals

when incorporated into E. coli. If polyoma virus,

which is the most infective tumor virus known

for hamsters, cannot cause tumors in the rDNA

state in E. coli, it is unlikely that other tumor

viruses will do so. This conclusion has had widespread, but not unanimous, acceptance. It has

been argued that there might be “something

special” about polyoma that prevents it from

causing tumors in this altered state; other

tumor viruses might still be able to do so. At one

meeting of RAC, in fact, it was suggested that

experiments with several other viruses be carried out to confirm the generality of the finding.

But how many more viruses? What is enough?

‘M. A. Israel, H. W. Chan, W. P. Rowe, and M. A. Martin, “Molecular Cloning of Polyoma Virus DNA in Eacherichki Cofi: Plasmid

Vector Systems,” Science 203:883-887, 1979.

*Some combinations of free plasmid and tumor virus DNA did

cause infections.
202 “ Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

For some, one carefully planned experiment

using the most sensitive tests is sufficient to

allay fears. But for others, significant doubt

about safety remains, regardless of how many

viruses are examined. The criteria depend on

an individual’s perception of risk.

Many experiments carried out for purposes

other than risk assessment have provided

evidence that scenarios of doom or catastrophe

are highly unlikely. This is the general consensus of specialists, not only in molecular biology,

but in population genetics, microbiology, infectious diseases, epidemiology, and public health.

Experiments have revealed that the structure

of genes from higher organisms (plants and

animals) differ from those of bacteria. Consequently, those genes are unlikely to be expressed accidentally by a bacterium; the original

fears of “shotgun” experiments have become

less well-founded. Hence, data gathered to date

have made the accidental construction of a new

epidemic strain more unlikely.

Conference discussions have also contributed

to a better understanding of the risks. At one

such conference,6 which was attended by 45 experts in infectious diseases and microbiology, it

was concluded that:

●

●

●

A

E. coli K-12 (the weakened form of E. coli,

used in experiments) does not flourish in

the intestinal tract of man;

the type of plasmid permitted by the Guidelines has not been shown to spread from E.

coli K-12 to other E. coli in the gut; and

E. coli K-12 cannot be converted to a harmful strain even after known virulence factors were transferred to it using standard

genetic techniques.

workshop sponsored by NIH7 provided a

forum for scientists to discuss the risks posed by

viruses in rDNA experiments. They concluded

that the risks were probably fess when a virus

was placed inside a bacterium in rDNA form

‘W’orkshop on Studies for Assessment of Potentiul Risks Associiit(?d With Recombinant DNA Kxperimenlal ion, ” k’iiln]outh, Miiss.,

June W-21 , 1977.

7’(Workshop to Assess Risks for Recomhinilnt DNA Experiments

Itlvoh’ing \’il’ill (k+nornes,” C(MpOIISOIWl h~ Iht? Niltiolliil Institutes

(d’ li[~illth ai~(i the Iiuropeiin Moleculi II . tliolo~v ol’~illliZillioll,”

AS(X)I , h;t~~liind, Jiin. 26-2&J, 1978.

than when it existed freely. * Experts in infectious disease have stressed repeatedly that the

ability of a micro-organism to cause disease

depends on a host of factors, all working together. Inserting a piece of DNA into a bacterium is

unlikely to suddenly transform the organism

into a virulent epidemic strain.

Careful calculations can also allay fears about

the damage a genetically engineered micro-organism might cause. Doomsday scenarios of

escaped E. coli that carry insulin or other

hormone-producing genes were recently examined in another workshop.8 Prior to this workshop, newspaper accounts raised the possibility

that an E. coli carrying the gene for human insulin production might colonize humans and

thus upset the hormonal balance of the body.

The participants calculated how much insulin

could be produced. First, it was assumed that a

series of highly unlikely events would occur—

accidental release, ingestion by humans, stable

colonization of the intestine by E. coli K-12. E.

coli constitutes approximately 1 percent of the

intestinal bacterial population, and it was

assumed that all the normal E. coli would be

replaced by the insulin-producing E. coli. Insulin

is made in the form of a precursor molecule,

proinsulin. It was assumed that 30 percent of all

bacterial protein production would be devoted

to this single protein, another highly unlikely

situation. If so, 30 micrograms (ug)-or 0.6

units—would then be made in the intestine.

Although proteins are very poorly absorbed

from the intestinal cavity, it was assumed for

the sake of argument that 100 percent of the

proinsulin would be absorbed into the circulation. Thus, 0.6 units of insulin would be added

to the normal daily human production of 25 to

30 units—an imperceptible difference.

Calculations like these have been carried out

for several other hormones. Even with the most

implausible series of events, leading to the

greatest opportunity for hormone production,

“(hi the other hiil)d, it hils heen iil’gue[i thiit this has provided

lriruses with ii new IXILIte for IIissell]illilti(]!l. Netw?rl ht?less, thel’e is

110 fn’idence thiit viruses Cilll readily escilpe tll)[ll the t)ilcleriil illld

Sul)sequellll.v CilUS(? intection.

“<Niilk)tliil lnslilute of Allergy iind infectious l)iseiises Workshop

011 Re[:onlhil~i]tlt DNA Risk AssessIIN?Ilt ,“ PilSildellill (Iillif., API..

11-12, 1980.
Ch. 10—The Question of Risk . 203

the conclusion is that normal hormone levels

would change by less than 10 percent. Similar

conditions for interferon production could

release approximately 70ug or the maximum

daily dose currently used in cancer therapy.

Long-term effects of such exposure are currently unknown; therefore, experiments using highproducing strains (106 molecules per cell or

more) are likely to be monitored if such strains

ever become available.

The NIH program of risk assessment, which

was formally started in 1979, continues to identify possible consequences of rDNA research.

Under the aegis of the National Institute of

Allergy and Infectious Diseases, the program

supports research studies designed to elucidate

the likelihood of harm. * In addition, it collates

general data from other experiments that might

be relevant to risk assessment. Other risk assessments are being conducted by European

organizations * * and by the U.S. Environmental

Protection Agency to assess the consequences of

releasing micro-organisms into the environment.

Thus far, there is no compelling evidence that

E. coli K-12 bacteria carrying rDNA will be more

hazardous than any of the micro-organisms

which served as the source of DNA. Nevertheless, all the experiments have dealt with one

genus of bacterium. Unless the conclusions

about E. coli can be extended to other organisms

likely to be used in experiments (such as Bacillus

subtilis and yeast), other assessments maybe appropriate.

“Extramural efforts were first conceived in the summer of 1975

to develop and test safer host-vector systems based on E. coli, the

interagency agreement entered into with the Naval Biosciences

Laboratory tested E. coli systems in a series of simulated

accidental spills in the laboratory. At the University of Michigan

the survival of these systems was tested in mice and in cultural

conditions simulating the mouse gastrointestinal tract. Tufts

University tested these systems in both mice and human

volunteers. Finally, the survival of host-vector systems in sewage

treatment plants was tested at the University of Texas. The peak

year for costs of supporting research contracts was 1978; over a

half-million dollars were required. currently, the cost of

maintaining the high containment facility at Frederick, Md., is

between $200,000 and $250,000 annually.

● ● First Report to the Committee on Genetic Experimentation , a

scientific committee of the International Council of Scientific

Unions, from the Working Group on Risk Assessment, July 1978.

Perception of risk

The probability of damage can be estimated

for various events. The entire insurance industry is based on the fact that unfavorable

events occur on a regular basis. The number of

people dying annually from cancer, or automobile accidents, or homicides can be predicted

fairly accurately. These estimates depend on

the availability of data and the assumptions that

the major determinants do not change from

year to year.

But even if the probability of damage is fairly

well known, a gap often exists between this

“real” probability of occurrence and the “perceived” probability. Two factors that tend to affect perceptions are the magnitude of the possible damage and the lack of individual control

over exposure to the risk. Both of these are significant factors in the fears associated with

rDNA and the manipulation of genes. Because

intuitive evaluations can contradict analytical

evaluations, the question of risk cannot be resolved strictly on an analytical basis. Its resolution will have to come through the political

process.

BURDEN OF PROOF

The possibility of inadvertently creating a

dangerous organism does exist, but its probability is lower than was originally thought.

Nevertheless, an important principle emerges

from the debate. Society must decide whether

the burden of proof rests with those who demand evidence of safety or with those who demand evidence of hazard. The former would

halt experiments until they are proved safe. The

latter would continue experiments until it is

shown that they might cause harm.

A significant theoretical difference exists between the two approaches. Evidence can almost

always be provided to show that something

causes harm—e.g., it can be demonstrated that a

poliovirus causes paralysis, that a Pneumococcus

causes pneumonia, that a rhinovirus causes the

common cold. However, it cannot be demonstrated that a poliovirus can never cause the

common cold, It cannot be demonstrated that

rDNA molecules will never be harmful. It can
204 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

only be demonstrated that harmful events are level of uncertainty it is willing to accept.

unlikely. Hence, society must determine what

Other concerns

Concerns raised by industrial

applications

Originally concerns involved hazards that

might arise in the laboratory. Now that there

are industrial applications of genetic engineering, the concerns include:

. risks associated with the laboratory construction of new strains of organisms,

. risks associated with industrial production

or consumer use of the new strains, and

 risks associated with the products obtained

from the new strains.

Many similar considerations apply to the assessment of the first two kinds of risks. Unless

the organisms used in an industrial production

scheme are thoroughly characterized, conjectured fears about their ability to cause disease

will continue. Even with a recombinant organism that has a well-defined sequence of DNA, a

break in containment would leave its behavior

in the environment questionable. Experience

with substances such as asbestos gives rise to

fears that exposure to the new biological systems might also cause unforseen pathological

conditions at some future time.

Hazards associated with products raise different questions. The growing consensus in

Federal regulatory agencies appears to be that

these products should be assessed like all

others—e,g., human growth hormone (hGH)

produced by genetically engineered bacteria

should be tested for purity, chemical identity,

and biological activity just like hGH from human

pituitary glands. The possibility of product

variation due to mutation of the bacteria,

however, suggests that batch testing and certification might be warranted as well. (For further

discussion see ch. 11.)

Concerns raised by the implications of

the rDNA controversy for general

microbiology

Questions about the potential harm from

genetically engineered micro-organisms have

led to questions about the efforts currently

employed to protect the public from work being

done with micro-organisms known to be hazardous. These viruses, bacteria, and fungi are

handled daily in laboratory experiments, in the

routine isolation of infectious agents from patients, and in the production of vaccines in the

pharmaceutical industry.

Questions have been raised about the efficacy

of regulations established for these various

potentially hazardous agents. A full-scale assessment is not within the scope of this study, but it

is clear that the questions are pertinent. Two

conclusions have been reached.

First, there is a growing belief that the mere

existence of a classification scheme for hazardous agents by the Center for Disease Control

(CDC) is not enough to ensure their safe handling. The Subcommittee on Arbovirus Laboratory Safety was formed recently because of concerns expressed in academic circles. Representatives from universities, the Public Health Service, the U. S. Department of Agriculture, and

the military, who constituted the subcommittee, are preparing a report based on an international survey of laboratory practices and infections. They found wide variation in the ways

different agents were handled. Most of their

recommendations are identical with those applicable to rDNA–that appropriate containment

levels be used with different viruses, that the

health of workers be monitored, and that an Institutional Biosafety Committee be appointed to

serve each institution.
Ch. 10—The Question of Risk . 205

Second, little is known about the health

record of workers involved in the fermentation

and vaccine industries. For most industrial

operations the evidence of harm is almost entirely anecdotal. Most industrial fermentations

are regarded as harmless; representatives of industry characterize it as a “non-problem” that

has never merited monitoring. Comprehensive

information on the potential harmful effects

associated with research using rDNA-carrying

. micro-organisms will not be available because

the Guidelines consider it the responsibility of

each institution or company to “determine, in

connection with each project, the necessity for

medical surveillance of recombinant-DNA research personnel. ” Hence some institutions

might decide to keep records of some or all activities; others might not.

To be sure, some companies have exceeded

the minimal medical standards set by NIH for

fermentation using rDNA-carrying micro-organisms–e.g., Eli Lilly & Co. requires that all

illnesses be reported to supervisors and that any

employees who are ill for more than 5 days

must report to a physician before being allowed

to return to work. Any employee taking antibiotics (which might make it easier for bacteria to

colonize) is restricted from areas where rDNA

research is being done until 5 days after the discontinuance of the antibiotic. At Abbott Laboratories, a physician checks into the illness of any

recombinant worker who is off more than 1

day–a precaution taken only after 5 days off

for workers in other areas. Lilly maintains a

computer listing of all workers involved in

rDNA activities. Lilly, the Upjohn Co., and

Merck, Sharp and Dohme have been in the

process of computerizing the health records of

all their employees over the past several years.

Work with rDNA has focused attention on

biohazards and medical surveillance—an awareness that had arisen in the past but had not been

sustained. * Consequently, several documents

on the subject either have been or will be published:

● As of September 1980, the National Institutes of occupational

Safety and Health and the Ent’ironnlental Protection Agency were

planning to fund assessments of the adequacy of current medical

surveillance technology,

●

●

●

●

CDC is preparing a complete revision of its

laboratory safety manual, which is widely

used as a starting point by other laboratories.

The Classification of Etiologic Agents on the

Basis of Hazard, which was last revised in

1974, has been expanded by CDC in collaboration with NIH into a Proposed Biosafety

Guidelines for Microbiological and Biomedical Laboratories. These guidelines serve the

purpose fulfilled by the Dangerous Pathogens Advisory Group (DPAG) in the United

Kingdom, although they lack any regulatory strength.

A comprehensive program in safety,

health, and environmental protection was

developed in 1979 by and for NIH. It is administered by the Division of Safety, which

includes programs in radiation safety, occupational safety and health, environmental protection, and occupational medicine.

The Office of Biohazard Safety, National

Cancer Institute has just completed a 3-

year study of the medical surveillance programs of its contractors; a report is being

drafted.

Although the academic, governmental, and

industrial communities have shown growing interest in biosafety, * no Federal agency regulates

the possession or use of micro-organisms except

for those highly pathogenic to animals and for

interstate transport. * * Whether such regulations are necessary is an issue that extends beyond the scope of this study. Nevertheless,

other countries—for instance the United Kingdom, with its DPAG—have acted on the issue.

This organization functions specifically to guard

against hazardous micro-organisms, by monitoring and licensing university and industrial

laboratories and meting out penalties when

necessary.

“Curiously, there is no formal society or journal, hut there has

been an annual Biological Safety Conference since 1955, conducted on a round-rohin hasis primarily h~ close associates of the

late Arnold G. Wedum, M. D.—fornler Director of Industrial Health

anci Safet-v at the LI. S. Army Bioiogica] Research Laboratories, Fort

Detrick, Md., who is regarded as the “Father of Microbiological

Safety. ”

● “In some States and cities, licensing is required for all facilities

handling pathogenic micro-organisms.
206 ● Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

Concerns raised by the implications of outside the cell. If the DNA is combined within

the rDNA controversy for other living cells, the Guidelines do not pertain. Figure

genetic manipulation 35 shows several methods that achieve the same

goal–transfering genetic material from one cell

Altering the hereditary characteristics of an to another, bypassing the normal sexual

organism-by using

several methods of

definition of rDNA

combination of the

rDNA is just one of the mechanisms of mating. It is particularly signifigenetic engineering. The cant that DNA from different species can be

refers specifically to the combined by all these mechanisms, only one of

DNA from two organisms which is rDNA. Different species of bacteria,

Figure 37.-Alternative Methods for Transferring DNA From One Cell to Another

A. The two cells are fused in toto

B. A microcell with a fragmented nucleus carries the DNA

C. Free DNA can enter the recipient cell in a number of ways: by direct microinjection, through

calcium-mediated transformation, or by being coated with a phospholipid membrane in

order to fuse with the recipient cell

D. The free DNA can be joined to a plasmid and transferred as recombinant DNA

SOURCE: Office of Technology Assessment.
Ch. 10—The Question of Risk ● 207

fungi, and higher organisms can all be fused or

manipulated. *

Opponents of rDNA have stated that combining genes from different species may disturb an

extremely intricate ecological interaction that is

only dimly understood. Hence, such experiments, it is argued, are unpredictable and therefore hazardous. If so, all the other methods

represented in figure 35 should be included in

the Guidelines. Yet they are not.

The most acceptable explanation for this inconsistency is that rDNA is currently the most

● For example, antibiotic resistant plasmids have been transferred from Staphylococcus aureus to Bacillus subtilis across

species barriers by transformation, not by rDNA. Foreign genes

for the enzyme amylase have also been introduced into B. subtilis.

Conclusion

Thus far, no demonstrable harm associated

with genetic engineering, and particularly

rDNA, has been found, But although demonstrable harm is based on evidence that damage has

occurred at one time or another, it does not

mean that damage cannot occur.

Conjectural hazards based on analogies and

scenarios have been addressed and most have

proved less worrisome than previously assumed. Nevertheless, there is agreement that

certain experiments, such as the transfer of

genes for known toxins or venoms into bacteria,

should still be prohibited because of the real

likelihood of danger. Still other experiments

cannot clearly be shown to be hazardous or

readily dismissed as harmless. Hence, a political

decision is likely to be required to establish

what constitutes acceptable proof and who

must provide it.

Given that potential harm can be identified in

some cases, its probable occurrence and magnitude quantified, and perceived risk taken into

account, a decision to proceed is usually based

on society’s willingness to take the risk. This

triad of the physical (actual risk), psychological

(perception of risk), and political (willingness to

take risk) plays a role in all decisions relating to

genetic engineering.

efficient and successful method of combining

genes from very diverse organisms. It is reasonable to ask, however, what would happen if any

of the other methods become equally successful. Will a profusion of guidelines appear? Will

one committee oversee all genetic experiments

Ethical and moral concerns

The perceived risk associated with genetic

engineering includes ethical and moral hazards

as well as physical ones. It is important to

recognize that these are part of the general

topic of risk. To some, there is just as much risk

to social values and structure as to human

health and the environment. (For further discussion see ch. 13.)

The potential benefits must always be considered along with the risks. Decisions made by

RAC have reflected this view—e.g., when it

approved the cloning of the genetic material of

the foot-and-mouth disease virus. The perceived

benefits to millions of animals outweighed the

potential hazard.

Recombinant DNA techniques represent just

one of several methods to join fragments of

DNA from different organisms. The current

Guidelines do no extend to these other techniques, although they share some of the same

uncertainties. Ignoring the consequences of the

other technologies might be viewed as an inconsistency in policy.

While the initial concerns about the possibility of hazards at the laboratory level appear to

have been overstated, other types of potential

hazards at different stages of the technology

have been identified. Emphasis has shifted

somewhat from conjectured hazards that might

arise from research and development to those

that might be associated with production technologies. As a consequence, there is a clearer

mandate for existing Federal regulatory agencies to play a role in ensuring safety in industrial

settings.
Chapter 11

Regulation of

Genetic Engineering
Chapter 11

Page

[introduction . . , . . . . . , . . . . , . . . . , . . . . . . . . , 211

Framework for the Analysis. . . . . . . . . . . . . . .211

Current Regulation: the NIH Guidelines. . . . . . . , 212

Substantive Requirements . . . . . . . . . . . . . . . .212

Administration . . . . . . . . , . . . . . . . . . . . . . .212

Provisions for Voluntary Compliance . . , . . .215

Evaluation of the Guidelines . , . . . , . . . . . , . . . 216

The Problem of Risk . ..................216

The Decisionmaking Process. . . . . . . . . . . . .221

Conclusion. . . . . . . . . . . . . . . . . . . . . . . . . . . .223

other Means of Regulation . . . . . . . . . . . . . . . . ..224

Federal Statutes. . . . . . . . . . . . . . . . . . . . . . . .224

“T’ort Law and Workmen’s Compensation . . , . .227

State and Local Law . . . . . . . . . . . . . . . . . . . . , 229

Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 230

Issue and options . . . . . . . . . . . . . . . . . . . . . . . . 230

Tables

Table No. Pag e

35. Containment Recommended by National

Institutes of Health . . . . . . . . . . . . . . . . . . . .,213

36. Statutes That Will Be Most Applicable? to

Commercial Genetic Engineering. . . . . . . . . . 224
Chapter 11

Regulation of Genetic Engineering

Introduction

Although no evidence exists that any harmful

organism has been created by molecular genetic

techniques, most experts believe that some

risk* is associated with genetic engineering.

One kind is relatively certain and quantifiable—

that of working with known toxins or pathogens. Another is uncertain and hypothetical–

that of the possible creation of a pathogenic or

otherwise undesirable organism by reshuffling

genes thought to be harmless. These may be

thought of as physical risks because they concern human health or the environment.

Concern has also arisen about the possible

long-range impacts of the techniques—that they

may eventually be used on humans in some

morally unacceptable manner or may change

fundamental views of what it means to be human. These possibilities may be thought of as

cultural risks, since they threaten fundamental

beliefs and value systems. 1

The issue of whether or not to regulate

molecular genetic techniques—and if so, to

what extent—defies a simple solution. Perceptions of the nature, magnitude, and acceptability of the risks differ drastically. Approximately

6 years ago, when the scientific community itself accepted a moratorium on certain classes of

recombinant DNA (rDNA) research, some scientists considered the concern unnecessary. Today, even though the physical risks of rDNA research are generally considered to be less than

originally feared—and the realization of its

benefits much closer–some people would still

prohibit it.

The Federal Government’s approach to this

issue has been the promulgation of the Guidelines for Research Involving Recombinant DNA

Molecules (Guidelines), by the National Institutes of Health (NIH). (See app. III-C for information about what other countries have done

● As used in this chapter, risk means the possibility of harm. ‘I-he

probability ot that harm occurring may he extremely low anckr

highly uncertain.

‘H. “Ilistam, ftngelhardt, Jr., ‘Taking Risks: Some Background

Issues in the I)etxite (k)ncerning Recombinant DNA Research,

Southern (;al(~ornia Law Review .51:6, pp. 1141-1151, 1978.

with respect to guidelines for rDNA.) Three

other available modes of oversight or regulation

are current Federal statutes, tort law, and State

and local law.

Framework for the analysis

In deciding how to address the risks posed by

genetic engineering, some of the important

questions that need to be examined are:

. How broadly the scope of the issue (or

problem) should be defined.

—Who identifies the risks and their magnitude?

—Who proposes the means for addressing

the problem?

● The nature of the procedural, decisionmaking mechanism.

—Who decides?

—Who will benefit from the proposed action and who will bear the risk?

—Will the risk be borne voluntarily or involuntarily?

—Who has the burden of proof?

—Should a risk/benefit analysis, or some

other approach, be used?

● The available solutions and their adequacy.

—Should there be full regulation, no regulation, or something in-between?

—What actions and actors should be cov-‘

ered?

—What is the appropriate means for enforcing a regulatory decision?

–Which agency or other group should do

the regulating?

Underlying these questions is the proposition,

widely accepted by commentators on science

policy, that scientists are qualified to assess

physical risk, since that involves measuring and

evaluating technical data. However, a judgment

of safety (the acceptability of that risk) can only

be made by society through the political process, since it involves weighing and choosing

among values. z 3456 Scientists are not nec-

‘William W. I,owrance, Of Aceep(able Risk: tkience and the Determination of &fety (l,os Altos, (hlit.: wi]lianl Kaufmann, ]n(~.,

1976).

211
212 . impacts of Applied Genetics—Micro-Organisms, Plants, and An/reals

essarily considered to be more qualified to make high value they place on unrestricted research

decisions concerning social values than other and because of possible conflicts of interest.

well-informed persons; they may in fact be less Moreover, according to this view, if society is to

qualified when the decision involves possible bear a risk, it should judge the acceptability of

restrictions on scientific research because of the that risk and give its informed consent to it.7

*

((:f)ntilll:~!fl,f}olll p. 211)

‘tAlvin W. Wcinlwrg, “Scienct~ illl[l ‘1’l’illlS-SCit? ll(X?, ” A4inerva,

10:2, April 1972. 7Engelhardt, op.cit.; Lowrance, op.cit.; and Bazelon, op. cit.

4Allan Mazur, ‘(Disputes Between Kxperts, ” Minerva 11:2, April ● [n practice, it may often he difficult to keep the two kinds of

1973. decisions separate, since the values of individual scientists may in-

‘A1.thur Kantrowitz, ‘

t

’r’he Science Court Experiment, ” Juri- tluence their interpretation of technical data, and since policymetrics ./ourna/, vol. 17, 1977, p. 332. makers may not have the technical competence to understand the

‘David L. Bazelon, “Risk and Responsibility, ” Science, vol. 205, risks sufficiently.e

July 20, 1979, pp. 277-280. ‘Weinberg, op. cit.; and Bazelon, op. cit.

Current regulation: the NIH Guidelines

The Guidelines have been developing in

stages over a period of approximately 6 years as

scientists and policymakers have grappled with

the risks posed by rDNA techniques. (This history, discussed in app. III-A, is crucial to understanding current regulatory issues, and it serves

as a basis for evaluating the Guidelines.) They

represent the only Federal oversight mechanism that specifically addresses genetic engineering.

Substantive requirements

The Guidelines apply to all research involving

rDNA molecules in the United States or its territories conducted at or sponsored by any institution receiving any support for rDNA research from NIH. Six types of experiments are

specifically prohibited: 1) the formation of

rDNA derived from certain pathogenic organisms; 2) the formation of rDNA containing

genes that make vertebrate toxins; 3) the use of

the rDNA techniques to create certain plant

pathogens; 4) transference of drug resistance

traits to micro-organisms that cause disease in

humans, animals, or plants; 5) the deliberate

release of any organism containing rDNA into

the environment; and 6) experiments using

more than 10 liters (1) of culture unless the

rDNA is “rigorously characterized and the

absence of harmful sequences established. ” A

procedure is specified for obtaining exceptions

from these prohibitions. Five types of experiments are completely exempt.

Those experiments that are neither prohibited nor exempt must be carried on in accordance with physical and biological containment levels that relate to the degree of potential

hazard. (See table 35.) Physical containment requires methods and equipment that lessen the

chances that a recombinant organism might escape. Four levels, designated P1 for the least

restrictive through P4 for the most, are defined.

Biological containment requires working with

weakened organisms that are unlikely to survive any escape from the laboratory. Three

levels are specified. Classes of permitted experiments are assigned both physical and biological containment levels. Most experiments

using Escherichia coli K-12, the standard laboratory bacterium used in approximately 80 percent of all experiments covered by the Guidelines, may be performed at the lowest containment levels.

ADMINISTRATION

The Guidelines provide an administrative

framework for implementation that specifies

the roles and responsibilities of the scientists,

their institutions, and the Federal Government.

The parties who are crucial to the effective

operation of the system are: 1) the Director of

NIH, 2) the NIH Recombinant DNA Advisory

Committee (RAC), 3) the NIH Office of Recombi-
Ch. 11—Regulation of Genetic Engineering ● 213

Table 35.–Containment Recommended by

National Institutes of Health

Biological—Any combination of vector and host must be

chosen to minimize both the survival of the system

outside of the laboratory and the transmission of the

vector to nonlaboratory hosts. There are three levels

of biological containment:

HV1 – Requires the use of Escherichia coli K12 or

other weakened strains of micro-organisms that

are less able to live outside the laboratory.

HV2– Requires the use of specially engineered strains

that are especially sensitive to ultraviolet light,

detergents, and the absence of certain

uncommon chemical compounds.

HV3 – No organism has yet been developed that can

qualify as HV3.

Physical—Special laboratories (P1-P4)

P1— Good laboratory procedures, trained personnel,

wastes decontaminated.

P2— Biohazards sign, no public access, autoclave in

building, hand-washing facility.

P 3 – Negative pressure, filters in vacuum line, class II

safety cabinets.

P 4 – Monolithic construction, air locks, all air

decontaminated, autoclave in room, all

experiments in class Ill safety cabinets (glove

box), shower room.

SOURCE: Office of Technology Assessment.

nant DNA Activities (ORDA), 4) the Federal Interagency Advisory Committee on Recombinant

DNA Research (Interagency Committee), 5) the

Institution where the research is conducted, 6)

the Institutional Biosafety Committee (IBC), 7)

the Principal Investigator (PI), and 8) the Biological Safety Officer.

The Director of NIH carries the primary burden for the Federal Government’s oversight of

rDNA activities, since he is responsible for implementing and interpreting the Guidelines, establishing and maintaining RAC (a technical advisory committee) and ORDA (whose functions

are purely administrative), and maintaining the

Interagency Committee (which coordinates all

Federal activities relating to rDNA). Under this

arrangement, all decisions and actions are taken

by the Director or his staff. For major actions,

the Director must seek the advice of RAC, and

he must provide the public and other Federal

agencies with at least 30 days to comment on

proposed actions. Such actions include: 1)

assigning and changing containment levels for

experiments, 2) certifying new host-vector systems, 3) maintaining a list of rDNA molecules exempt from the Guidelines, 4) permitting exceptions to prohibited experiments, and 5) adopting

changes in the Guidelines.

For other specified actions, the Director need

only inform RAC, the IBCs, and the public of his

decision. The most important of these are: 1)

making minor interpretive decisions on containment for certain experiments; 2) authorizing,

under procedures specified by RAC, large-scale

work (involving more than 101 of culture) with

rDNA that is rigorously characterized and free

of harmful sequences; and 3) supporting laboratory safety training programs. Every action

taken by the Director pursuant to the Guidelines must present ‘(no significant risk to health

or the environment. ”

RAC is an advisory committee to the Director

on technical matters. It meets quarterly. Its purpose, as described in its current charter of June

26, 1980 (and unchanged since its inception in

October 1974), is as follows:

The goal of the Committee is to investigate

the current state of knowledge and technology

regarding DNA recombinant, their survival in

nature, and transferability to other organisms;

to recommend guidelines for the conduct of

recombinant DNA experiments; and to recommend programs to assess the possibility of

spread of specific DNA recombinant and the

possible hazards to public health and to the environment. This Committee is a technical committee, established to look at a specfic problem. (Emphasis added.)

The charter and the Guidelines also assign it

certain advisory functions that have changed

over time.

The RAC is composed of not more than 25

members. At least eight must specialize in molecular biology or related fields; at least six must

be authorities from other scientific disciplines;

and at least six must be authorities on law,

public policy, the environment, public or occupational health, or related fields. In addition,

76-565 0 - 81 - 15
214 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

representatives from various Federal agencies

serve as nonvoting members.

ORDA performs administrative functions,

which include reviewing and approving IBC

membership and serving as a national center

for information and advice on the Guidelines

and rDNA activities.

The Interagency Committee was established in

October 1976 to advise the Secretary of the then

Department of Health Education and Welfare

(HEW) [now Health and Human Services

(DHHS)] and the Director of NIH on the coordination of all Federal activities relating to

rDNA. It has thus far produced two reports. Its

first, in March 1977, concluded that existing

Federal law would not permit the regulation of

all rDNA research in the United States to the extent considered necessary and recommended

new legislation, specifying the elements of that

legislation. ’” The second, in November 1977,

surveyed international activities on regulating

the research and concluded that, while appropriate Federal agencies should continue to work

closely with the various international organizations, no formal governmental action was necessary to produce international control by means

of a treaty or convention.11 It is currently considering issues arising from the large-scale industrial applications of rDNA techniques.

Under the Guidelines, essentially all the responsibility for overseeing rDNA “experiments

lies with those sponsoring or conducting the research. The Institution must implement general

safety policies, * establish an IBC, which meets

specified requirements, and appoint a Biological

Safety Officer. The Biological Safety Officer, who

is needed only if the Institution conducts experiments requiring P3 or P4 containment, (see

table 35) oversees safety standards. The initial

responsibility for particular experiments lies

‘Interim Report of the Federal Interagency Committee on Recombinant DNA Research: Su,gested Elements-for Legislation, Mar. 15,

1977, pp. 9-10.

IDlbid., pp. 11-15,

I I Report of the Federal intet-~ency Committee on Recombinant

DNA ftesearch: International Activities, Novendxx 1977, pp. 13-15.

“These include conducting imy heid[h surveillance that it determines to be necessary and ensuring iippr~priiiie tritining for the

IBC, Biological Safety officers, Principal lnv{?sti~iit~rs, and laboratory staff.

with the PI, the scientist receiving the funding.

This person is responsible for determining and

implementing containment and other safeguards and training and supervising staff. In addition, the PI must also submit a registration

document that contains information about the

project to the IBC, and petition NIH for: 1) certification of host-vector systems, 2) exceptions

or exemptions from the Guidelines, 3) and determination of containment levels for experiments not covered by the Guidelines. Furthermore, all of the above have certain reporting requirements designed so that ORDA is eventually

informed of significant problems, accidents, violations, or illnesses.**

The IBC is designed to provide a quasi-independent review of rDNA work done at an institution. It is responsible for: 1) reviewing all

rDNA research conducted at or sponsored by

the institution and approving those projects in

conformity with the Guidelines; 2) periodically

reviewing ongoing projects; 3) adopting emergency plans for spills and contamination; 4)

lowering containment levels for certain rDNA

and recombinant organisms in which the absence of harmful sequences has been established; and 5) reporting significant problems,

violations, illnesses, or accidents to ORDA

within 30 days.*** The IBC must be comprised

of no fewer than five members who can collectively assess the risks to health or the environment from the experiments. At least 20

percent of the membership must not be otherwise affiliated with the institution where the

work is being done, and must represent the interests of the surrounding community in protecting health and the environment. Committee members cannot review a project in

which they have been, or expect to be, involved

or have a direct financial interest. Finally, the

Guidelines suggest that IBC meetings be public;

minutes of the meetings and submitted documents must be available to the public on

request.

● ● The PI is required to report this information within 30 days to

ORDA and his IBC. The Biological Siifet~ officer musl report the

same to the Institution and the IBC unless the PI has clone so. The

institution must report within 30 dii~s to ORDA unless the PI or

IB(: has done so.

*” “It does not have to report if the PI hits done SO.
Ch. n-Regulation of Genetic Engineering ● 215

The requirements imposed on an institution

and its scientists are enforced by the authority

of NIH to suspend, terminate, or place other

conditions on its funding of the offending projects or all projects at the institution. Compliance

is monitored through the requirements for notification mentioned above.

PROVISIONS FOR VOLUNTARY COMPLIANCE

Organizations or individuals who do not receive any NIH funds for rDNA research are not

covered by the Guidelines. These include other

Federal agencies, institutions and individuals

funded by those agencies, and corporations.

Federal agencies other than NIH that conduct

or fund rDNA research have proclaimed their

voluntary compliance with the Guidelines. *

Staff scientists have been so informed by memoranda. As for outside investigators, this policy

has been implemented through the grant application process. Instructions in grants applications contain policy statements regarding

compliance with the Guidelines, and applicants

are sometimes contacted to ascertain their

knowledge of the Guidelines. Information has

been requested for certain experiments, and

IBC membership has been reviewed. From time

to time, the agencies have consulted with NIH

on matters that need interpretation.

Part VI of the Guidelines is designed to encourage voluntary compliance by industry. It

creates a parallel system of project review and

IBC approval analogous to that required for

NIH-funded projects, modified to alleviate industry’s concerns about protection of proprietary information.

The Freedom of Information Act requires

Federal agencies, with certain exceptions, to

make their records available to the public on request.One of the exceptions is for trade secrets

and proprietary information obtained from

others. Part VI contains several provisions for

protecting this information. Perhaps the most

important is a process whereby a corporation

*These agencies are the National Science Foundation, the Department of Agriculture, the Department of Energy, the Veterans

dministration, and the Center for Disease Control. Two other

gencies, which have expressed interest in this research but are

ot currently sponsoring any projects, are the Department of De-

;nse and the National Aeronautics and Space Administration.

may request a presubmission review of the

records needed to register its projects with NIH.

The DHHS Freedom of Information Officer

makes an informal determination of whether

the records would have to be released. If they

are determined to be releasable, the records are

returned to the submitting company. The

Guidelines also require that NIH consult with

any institution applying for an exemption,

exception, or other approval about the content

of any public notice to be issued when the application involves proprietary information. As a

matter of practice, such applications are also

considered by RAC in nonpublic sessions.

Large-scale experiments (more than 10 1 of

culture) with rDNA molecules are prohibited

unless the rDNA is “rigorously characterized

and the absence of harmful sequences established.” Such experiments are actually scale-ups

of potential industrial processes. Those meeting

this standard may be approved by the Director

of NIH under procedures specified by RAC. * At

its September 1979 meeting, RAC adopted procedures for review that require the applicant to

submit information on its laboratory practices

and containment equipment. Subsequently, recommendations were developed for large-scale

uses of organisms containing rDNA. These were

published in the Federal Register on April 11,

1980. Besides setting large-scale containment

levels, they require the institution to appoint a

Biological Safety Officer with specified duties,

and to establish a worker health surveillance

program for work requiring P3 containment. At

its September 1980 meeting, RAC modified its

review procedures so that the application need

only specify the large-scale containment level at

which the work would be done, without providing details on containment equipment. RAC

will continue to review the biological aspects of

the applications in order to determine that

rDNA is rigorously characterized, that the absence of harmful sequences is established, and

that the proposed containment is at the appropriate level.

“It is NIH, not the company proposing the scale-up, that determines if the rDNA to be used is “rigorously characterized and the

absence of harmful sequences established.”. *2

Iz(;uide]ines for Research [nvolving Recombinant DNA Molecules, sec. lk’-E-l-b-(3)-(d).
216 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Evaluation of the Guidelines

Two basic issues must be addressed. The first

is how well the Guidelines confront the risks

from genetic engineering, which may not have a

definitive answer in view of the uncertainty

associated with most of the risks. Consequently,

it is also necessary to consider a second issue—

whether confidence is warranted in the decisionmaking process responsible for the Guidelines.

THE PROBLEM OF RISK

The Guidelines are designed to address the

risks to public health and the environment from

either rDNA molecules or organisms and viruses containing them. The underlying premise

is that research should not be unreasonably

restricted. This is essentially a risk-benefit approach; at the time that the original Guidelines

were drafted, it represented a compromise between the extremes of no regulation and of no

research without proof of safety. physical and

biological containment levels were established

for various experiments based on estimated

degrees of risk. The administrative mechanism

created by the Guidelines is that of a Federal

agency —NIH—advised by a diverse body of

experts— RAC. Scientific advice on the technical

aspects of risk assessment is provided by technical experts on RAC; public input is provided by

experts in nontechnical subjects and by the

right of the public to comment on major actions,

which are published in the Federal Register.

Compliance is accomplished by a combination

of local self-regulation and limited Federal oversight, with the ultimate enforcement resting in

the Federal funding power.

Since their initial appearance, the Guidelines

have evolved. As scientists learned more about

rDNA and molecular genetics, two trends occurred. First, containment levels were progressively lowered. Major revisions were made in

1978 and 1980; minor revisions were often

made quarterly, as proposals were submitted to

the RAC at its quarterly meetings, recommended by RAC, and accepted by the Director.

By now, approximately 85 percent of the permitted experiments can be done at the lowest

physical and biological containment levels. Second, the degree of centralized Federal oversight has been substantially reduced to the point

where almost none remains. Under the 1976

Guidelines, all permitted experiments ultimately

had to be reviewed by the IBC and ORDA before

they could be started; the 1978 Guidelines no

longer required preinitiation review of most

experiments by ORDA, although ORDA continued to maintain a registry of experiments

and to review IBC decisions. Under the

November 1980 revision to the Guidelines, there

will be no Federal registration or review of experiments for which containment levels are

specified in the Guidelines. About 97 percent of

the permitted experiments fall into this

category.

Preinitiation review of experiments by RAC

has been an important part of the oversight

mechanism. Expert review encourages experimental design to be well thought out and provides a means for catching potential problems,

e.g., one application reviewed by RAC never

mentioned that the species to be used as a DNA

donor was capable of manufacturing a potent

neurotoxin; it was turned down after a RAC

member familiar with the species brought this

fact to the Committee’s attention.13

The burdens imposed on rDNA activities by

the Guidelines appear to be reasonable in view

of continuing concerns about risk. Less than 15

percent of permitted experiments require preinitiation approval by the local IBC’s, which usually meet monthly. Preinitiation approval of experiments by NIH is required only for: 1) experiments that have not been assigned containment

levels by the Guidelines; 2) experiments using

new host-vector systems, which must be certified by NIH; 3) certain experiments requiring

case-by-case approval; and 4) requests for exceptions from Guideline requirements. The lowest containment levels place minimal burdens

on the experimenter. (see table 35). For industrial applications, NIH approval must be

received not only when the project is scaled-up

beyond the 10-1 limit, but also for each addi

tional scale-up of the same project. Many repre

sentatives of industry consider these subse

‘3R. M. Henig, “Trouble on the RAC—Committee Splits Ove

Downgrading of E. co/i Containment,” Bioscience, vol. 29, pp. 759

762, December 1979.
Ch. n-Regulation of Genetic Engineering ● 217

quent approvals to be unnecessary and burdensome.

Information about whether the Guidelines

have been a disadvantage for U.S. companies in

international competition is scanty. Examples

include the approximately l-year headstart two

European groups were given while the cloning

of hepatitis B virus was prohibited, the advantage some European companies had in using

certain species of bacteria for cloning under

conditions that were prohibited in the United

States, and the delays some pharmaceutical

companies faced because they had to build better containment facilities.

The present Guidelines are a comprehensive,

flexible, and nonburdensome way of dealing

with the physical risks associated with rDNA research while permitting the work to go forward. That is all they were ever intended to do.

The Scope of the Guidelines.—In many

respects, the Guidelines do not address the full

scope of the risks of genetic engineering. They

cover one technique, albeit the most important;

they do not address the admittedly uncertain,

long-term cultural risks; they are not legally

binding on researchers receiving funds from

agencies other than NIH; and they are not binding on industry.

Other genetic techniques present risks similar to those posed by rDNA, but to a lesser degree. Recombinant DNA is the most versatile

and efficient technique; it uses the greatest

variety of genetic material from the widest

number of sources with reasonable assurance

of expression by the host cell. Cell fusion of

micro-organisms, which also involves the uncertain risk of recombining the genetic material of

different species, is significantly less versatile

and efficient than rDNA but mixes more genetic

material. In addition, the parental cells may contain partial viral genomes that could combine to

form a complete genome when the cells are

fused. Transformation, a technique known for

decades, similarly involves moving pieces of

DNA between different cells. However, it is significantly less versatile and efficient than cell fusion, and it is generally considered to be virtually risk-free. Thus, cell fusion is in a gray area

between the other two techniques; yet no risk

assessment has been done, and no Federal oversight exists.

Another limitation in the scope of the guidelines—and in the process by which they were

formulated—is that long-range cultural risks (as

distinguished from policy issues related to safety) were never addressed. As noted by the Director of NIH:14

NIH has been addressing the policy questions involving the safety of this research, not

the ‘potential future application . . . to the altering of the genetic character of higher forms of

life, including man’ . . .

Perhaps it was inappropriate to do more. Such

ethical issues might be considered premature in

view of the level of the development of the technology. The desire among many molecular biologists to move ahead with the research meant

that experiments were being done; therefore

the immediate potential for harm was to health

and the environment. Thus, it was arguably

necessary to develop a framework to deal with

the risks based on what was known at the time.

On the other hand, the broader questions of

where the research might eventually lead and

whether it should be done at all have been

raised in the public debate. They have not been

formally considered by the Federal Government.

Another limitation in the scope of the Guidelines is their nonapplicability to research

funded or performed by other Federal agencies.

However, agencies supporting such research

are complying with the Guidelines as a matter of

policy. There appears to be little reason for

questioning these declarations of general policy.

In practice, problems might arise if a mission is

perceived to be at odds with the Guidelines or

because of simple bureaucratic defense of territory —e.g., when the 1976 Guidelines were promulgated, two agencies—the Department of Defense (DOD) and the National Science Foundation (NSF) preserved the right to deviate for reasons of national security or differing interpreta1443 F.R. 6o103, Dec. 22, 1978, citing 43 F.R. 33067, JuI.v 28,

1978.
218 ● lmpacts of Applied Genetics—Mlicro-Organisms, Plants, and Animals

tions, respectively. * DOD no longer claims an

exception for national security.l5 NSF took its

position when it approved funding for an experiment using a particular species of yeast that

had not been certified by NIH, relying on an ambiguously worded sectionl6 in the Guidelines to

assert that it could certify the host. subsequent

revisions explicitly stated that these hosts had to

be certified by the Director of NIH1718 a nd

removed many similar ambiguities.

In the final analysis, NIH has indirect leverage

over the actions of other agencies through its

funding. All non-NIH funded rDNA projects at

an institution which also receives NIH funds for

rDNA work must comply with the Guidelines;

otherwise NIH funds may be suspended or terminated.

While the procedures of other agencies for

administering compliance are significantly less

formal than those created by the Guidelines for

NIH, they do rely heavily on NIH for help and

advice, and they coordinate their efforts

through the Interagency Committee and their

nonvoting membership on RAC. So far, this voluntary compliance by the agencies appears to

be working fairly well.

The most significant limitation in the scope of

the Guidelines is their nonapplicability to industrial research or production on other than a

voluntary basis. This lack of legal authority

raises concerns not only about compliance but

also about NIH’s ability to implement a voluntary program effectively.

whether every company working with rDNA

will view voluntary compliance to be in its best

interest depends on a number of factors. In

the past, certain short-sighted actions by even a

few companies in a given industry has led to

*For a statement of the DOD position, see the minutes of the

November 23, 1976, meeting of the Federal Interagency Committee. At that time, DOD had no active or planned rDNA projects.

NSF’s statement of its intention to “preserve some level of independence of decision” was expressed in an internal NIH memorandum dated February 24, 1978, from the Deputy Director for

Science, NIH, to the Director, NIH.

lsDr. John H. Moxley, III., Assistant Secretary of Defense for

Health Affairs, personal communication, Nov. 18, 1980.

lo{’FUngal or similar Lower Eukaryotic Host-Vector Systems, ” 41

F.R. 27902, 27920, July 7, 1976.

1743 F.R. 60108,” Dec. 22, 1978, sec. 111.C.5 of the 1978 Guidelines.

1s45 F*R. 6724, Jan$ 29, 1980, ~~o 111.C.5 of the 1980 Guidelines.

well-documented abuses and a host of Federal

laws to curtail them. However, at least two constraints are operating in the case of the biotechnology industry. First, the possibility of tort

lawsuits is an inducement to comply with the

Guidelines, which would probably be accepted

as the standard of care against which alleged

negligence would be evaluated. (This concept is

discussed in greater detail in the section on Tort

Law and workman’s compensation.) second,

the threat of statutory regulation, which the

companies have sought to avoid, always exists.

Other factors are also at work. Except for the

10-1 limitation, for which cases-by-case exceptions must be sought, the large-scale containment recommendations of April 11, 1980, are

not excessively burdensome, at least for pharmaceutical companies. The requirements are

similar to measures that must currently be

taken to prevent product contamination. In addition, the public debate should have made each

company aware of the problems and the need

for voluntary compliance before it invested substantially in biotechnology; expensive controls

will not have to be retrofitted. However, one

definite concern is that new companies attracted to the field will perceive their interests

differently. Because they did not actually experience the period when legislation seemed inevitable and because they will be late entries in the

race, they may be inclined to take shortcuts.

Besides the concern about whether industry

has sufficient incentive to comply, there are a

number of other reasons for questioning the effectiveness of the voluntary program. First, until

very recently no member of RAC was an expert

in industrial fermentation technology-yet the

committee has been considering applications

from industry for large-scale production since

September 1979. * This drawback was demonstrated at its March 1980 meeting, when the

committee expressed uncertainty over what

Federal or State safety regulations presently

cover standard fermentation technology em-

● At its September 1980 meeting, RAC passed the following resolution, which has been accepted by the Director of NIH: 19

Members should he chosen to provide expertise in fermentation

techmlo~y, engineering, tind other wpcts of large-wide production.

A fermentation technology expert was appointed in January

1981.

]s45 F.R. 77373, Nov. 21, 1980.
Ch. 11-Regulation of Genetic Engineering . 219

ployed by the drug industry. Various members

expressed concern in the March and June 1980

meetings about the Committee’s continuance to

make recommendations on the applications

without a firm knowledge of large-scale production.

second, the provisions in part VI of the Guidelines, which allow prior review of submitted information by the DHHS Freedom of Information

Act Officer, give an industrial applicant the option of withholding potentially important information on the grounds of trade secrecy, even

when DHHS disagrees. Third, because some

RAC members have been opposed to discussing

industrial applications in closed session (needed

to protect proprietary information), they have

chosen not to participate in those sessions.

Thus, some diversity of opinion and expertise

has been lost. Fourth, monitoring for compliance after the scale-up applications are granted

is limited. Some early applications were granted

on the condition that NIH could inspect facilities, and at least one inspection was made.

Under procedures adopted at the September

1980 meeting, a company’s IBC will be responsible for determining whether the facilities meet

the standards for the large-scale containment

level assigned by RAC. A working group of RAC

may visit the companies and their IBCs from

time-to-time but only for information gathering

purposes, rather than for regulatory actions.

Fifth, even if noncompliance were found, no

penalties can be imposed.

The members of RAC, acutely aware of the

problems with voluntary compliance by industry, have been deliberating about them for

almost 2 years. At a meeting in May 1979, they

decided, by a vote of nine to six with six abstentions, to support the principle of mandatory

compliance with the Guidelines by non-NIH

funded institutions. However, the Secretary of

HEW (Joseph Califano) decided to continue with

the development of voluntary compliance provisions20 which were adopted as Part VI of the

Guidelines in January 1980. Actual RAC review

of submissions from the private sector for largescale work began in September 1979. At a meeting in June 1980, RAC debated the effectiveness

ZORAC minutes of sept. 6-7, 1979, p. 16, in Recombinant DNA Research, vol. 5, (Wash., D. C.: HEW, 1980), p. 165.

of NIH’s quasi-regulation of industry. A primary

concern was whether the RAC would be viewed

as giving a “stamp of approval” to industrial projects, when, in fact, it has neither the authority

nor the ability to do so. One member, lawyer

Patricia King, stated:2l

Voluntary compliance is the worst of all possible worlds. . . .You achieve none of the objectives of regulation and none of the benefits of

being unregulated. All you’re saying is ‘I give a

stamp of approval to what I see here before me

without any authority to do anything.’

Most of the speakers expressed the desire that

the various agencies in the Interagency Committee be responsible for such regulation. However, the Interagency Committee, which has

been studying the problem since January 1980,

has yet to decide what it can do. Thus, many of

its members see RAC as filling a regulatory void

until the traditional agencies take action.

Some regulatory agencies have begun to deal

with specific problems within their areas of interest. The Occupational Safety and Health Administration will decide its regulatory policy on

the basis of a study of potential risks to workers

posed by the industrial use of rDNA techniques

being conducted by the National Institute of

Occupational Safety and Health (NIOSH). In a

letter to the Director of NIH dated September

24, 1980, Dr. Eula Bingham, then Assistant Secretary for Occupational Safety and Health of the

Department of Labor, estimated this process

would take approximately 2 years. The Environmental Protection Agency (EPA) has awarded

several contracts and grants to assess the risks

of intentional release of genetically engineered

micro-organisms and plants into the environment. And the Food and Drug Administration

(FDA) has begun to develop policy with respect

to products made by processes using genetically

engineered micro-organisms. (Further details

on agency actions are discussed in the section,

Federal Statutes.)

Compliance.—The primary mechanism in

the Guidelines for enforcing compliance is local

self-regulation, with very limited Federal over21 Susan Wright, Recommended NA Policy: Controlling LargeScale Processing,” Environment, vol. 22, September 1980, pp.

29,32.
220 ● Impacts of Applied Genetlcs—Micro-Organisms, Plants, and Animals

sight. Penalties are based on NIH’s power to restrict or terminate its funding.

The initial responsibility for compliance lies

with the scientist doing the experiments. A researcher’s attitude toward the risks of rDNA

techniques and the necessity for the Guidelines

appear to be an influential factor in the degree

of compliance. A science writer who worked for

3 months in a university lab in 1976 noted sloppy procedures and a cavalier attitude, stating:

“Among the young graduate students and postdoctorates it seemed almost chic not to know

the NIH rules.”22 On the other hand, in the case

of a recent violation of the Guidelines, it appears

as if the investigator’s graduate students were

the first to raise questions.23 24 Competitiveness

is another important factor. Novice scientists

must establish reputations, secure tenure in a

tight job market, and obtain scarce research

funds; established researchers still compete for

grants and certainly for peer recognition. This

competitive pressure could provide strong incentives to bend the Guidelines; on the other

hand, it might be channeled to encourage compliance if it is believed that NIH will in fact

penalize violations by restricting or terminating

funding.

The first level of actual oversight occurs at

the institution. An argument can be made that

reliance on the PI and an IBC (that might be

composed mostly of the PI’s colleagues) provides

too great an opportunity for lax enforcement or

coverups. On the other hand, spreading responsibility among the institution, the PI, the IBC,

and, in the case of more hazardous experiments, the Biological Safety Officer might reduce the chance of violations being overlooked

or condoned. This responsibility is enhanced by

the reporting requirements borne by each of

these parties, designed so that ORDA learns of

“significant” problems, accidents, violations, and

illnesses. What is “significant” is not defined.

Public involvement at the local level acts as an

additional safeguard. Twenty percent of the

~ZJanet L. Hopson, “Recombinant Lab for DNA and MY 95 Days

in It,” Smithsonian, vol. 8, June 1977, p. 62.

Z3D0 Dickson, (, Another Vio]ation of NIH Guide] ines, j) ~ature vol.

286, Aug. 14, 1980, p. 649.

i~D. Dickson, “DNA Recombination Forces Resignation, ” ~tlfUJl?

vol. 287, Sept. 18, 1980, p. 179.

IBCs members must be unaffiliated with the institution. IBC documents, including minutes of

meetings, are publicly available, but meetings

are not required to be held in public. On the

other hand, the probable inability of the members who represent the public to understand

the technical matters might limit their effectiveness.

How successful has compliance been? Three

known violations have occurred. In each, no

threat to health and the environment existed. In

each, there was some confusion as to why the

violations occurred. NIH is presently investigating the third violation. For the first two, it

accepted explanations of misunderstandings

and misinterpretations of the Guidelines. However, a Senate oversight report concluded:25

While undoubtedly most researchers have

observed the guidelines conscientiously, it is

equally clear that others have substituted their

own judgments of safety for those of NIH.

No firm conclusions can be drawn on the question of compliance. The reporting of only a few

violations could be evidence that the compliance

mechanism embodied in the Guidelines has

been working well. Or it could mean that some

violations are not being discovered or reported.

The November 1980 amendments to the

Guidelines substantially changed procedures

designed to monitor compliance by abolishing a

document called a Memorandum of Understanding and Agreement (MUA). It had been required for 15 to 20 percent of all experiments,

those thought to be potentially most risky. The

MUA, which was to be filed with ORDA by an

institution, provided information about each experiment, and it was the institution’s certification to NIH that the experiment complied with

the Guidelines. By having the MUAs, ORDA

could monitor for inconsistencies in interpreting the Guidelines, actual noncompliance, and

the consistency and quality with which IBCs

functioned nationwide. The amendments continued a trend begun in January 1980, when approximately 80 percent of the experiments,

‘

s

’’ Recombinant DNA Research and Its Applications,” Oversight

Report, Subcommittee on Science, Technology, and Space of the

Senate Committee on Commerce, Science and Transportation,

Aug. 1978, p. 17.
Ch. 11-Regulation of Genetic Engineering . 221

those done with E. coli K-12, were exempted

from the MUA requirement.

The abolition of the MUA essentially abolished centralized Federal monitoring of rDNA

experiments. The only current Guideline provision that serves this kind of monitoring function

is the requirement that the institution, the IBC,

or the PI notify ORDA of any significant violations, accidents, or problems with interpretation. Limited monitoring of large-scale activities

continues. Under NIH procedures (which are

not part of the Guidelines) for reviewing applications for exemptions from the 10-1 limit, the

application must include a copy of the registration document filed with the IBC. The manufacturing facilities may also be inspected by NIH,

not for regulatory purposes, but to gather information for updating its recommended largescale containment levels; The abolition of the

MUA is consistent with traditional views that

Government should not interfere with basic scientific research. Whether or not it will reduce

either the incentive to comply with the Guidelines or the likelihood of discovering violations

remains to be seen.

THE DECISIONMAKING PROCESS

Another way to evaluate the Guidelines besides considering their substantive requirements is to look at the process by which they

were formulated. In a situation where there is

uncertainty and even strong disagreement

about the nature, scope, and magnitude of the

risks, it is difficult to judge whether or not a

proposed solution to a problem will be a good

one. Society’s confidence in the decisionmaking

process and in the decisionmakers then becomes the issue. As David L. Bazelon, Chief

Judge of the U. S. Court of Appeals for the District of Columbia, has stated:26

When the issues are controversial, any decision may fail to satisfy large portions of the community. But those who are dissatisfied with a

particular decision wil1 be more likely to acquiesce in it if they perceive that their views and

interests were given a fair hearing. If the decision-maker has frankly laid the competing considerations on the table, so that the public

knows the worst as well as the best, he is unlikeZ@D. L. Bazelon, ‘(coping With Technology Through the Legal

Process, ” 62 Cornell Law Review 817,825, June 1977.

ly to find himself accused of high-handedness,

deceit, or cover-up. We simply cannot afford to

deal with these vital issues in a manner that invites public cynicism and distrust.

The manner in which the Guidelines themselves evolved has been controversial. (For a

detailed discussion see app. III-A.) Initially, the

scope and nature of the problem was defined by

the scientific community; NIH organized RAC

along the lines suggested by the NAS committee

letter referred to in app. III-A. One of the goals

of RAC was to recommend guidelines for rDNA

experiments; it was not charged with considering broader ethical or policy issues or the fundamental question of whether the research should

have been permitted at all. The original Guidelines were produced by a committee having

only one nonscientist.

In late 1978, the Secretary of HEW significantly restructured RAC and modified the

Guidelines in order to increase the system’s

accountability to the public, to “provide the opportunity for those concerned to raise any

ethical issues posed by recombinant DNA research”{ and to make RAC “the principal advisory body . . . on recombinant DNA policy. ”27

However, it has remained in large part a technically oriented body. Its charter was not

changed in this respect; the Guidelines themselves state that its advice is “primarily scientific

and technical, ” and matters presented for its

consideration have continued to be mostly technical. One area where RAC has played a significant policy role, however, is in dealing with the

issue of voluntary compliance by industry.

It could be argued that the system did provide

for sufficient public input into the formulation

of the problem* and that no other formulation

was realistic. The two meetings in 1976 and

1977 of the NIH Director’s Advisory Committee

and the hearing chaired by the general counsel

of HEW in the fall of 1978 provided the opportunity for public comment on the overall FedZTJoSeph A. Ca]ifano, “Notice of Revised Guidelines—Recombinant DNA Research, ” 43 F.R. 60080-60081, Dec. 22, 1978.

● The problem was conceived in terms of how to permit the re.

search to be done while limiting the physical risks to an acceptable

level. Other formulations were possible, the broadest being how

to limit all risks, including cultural ones, to an acceptable level.

Such a formulation could have resulted in a prohibition of the

research.
222 . Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

eral approach to the controversy, including

whether or not the problem had been too narrowly phrased. similarly, Congress had the

opportunity in 1977 to reevaluate the entire

institutional response, taking into account any

moral objections to the research in addition to

those concerning safety. Yet the principal bills

were based on the proposition that the research

continue in a regulated fashion.

A related issue is the one of burden of proof.

Should the proponents of a potentially beneficial technology be required to demonstrate

minimal or acceptable risk even if that risk is

uncertain or even hypothetical? Or should its

opponents be required to demonstrate unacceptable risk? If the proposition is accepted that

those who bear the risks, in this case the public

as well as the scientists, must judge their acceptability, then the burden must be on the proponents. The scientific community clearly accepted this burden. The moratorium proposed

by the NAS committee in July 1974 called for a

suspension of certain types of rDNA experiments until the risks could be evaluated and

procedures for adequately dealing with those

risks could be developed. The Guidelines prohibited some experiments, specified containment levels for others, and required certification of host-vector systems. All actions approved

by the Director of NIH, including the lessening

of the restrictions imposed by the original

Guidelines, have had to meet the requirement of

presenting “no significant risk to health or the

environment. ”

Two other criticisms have been directed

against RAC, particularly in its early days. The

first concerned inherent conflicts of interest.

RAC’s members were drawn from molecular

biology and related fields. One of the early

drafts of the Guidelines was criticized as being

“tailored to fit particular experiments that are

already on the drawing boards. ”28 However,

only a few of the members were actually working with rDNA.29 A more serious criticism was

the lack of a broad range of expertise. Although

the risks had been expressed in terms of potential hazards to human health and the environment, the original RAC had no experts in the

areas of epidemiology, infectious diseases, botany or plant pathology, or occupational health.

It did have one expert in enteric organisms, E.

coli in particular.

These shortcomings were eventually remedied by expanding RAC’s membership to allow

the appointment of other experts, including

some from nontechnical fields such as law and

ethics. In addition to providing knowledge of

other fields, these members served as disinterested advisors, since they had no direct interest in expediting the research. Thus, the Government dealt with the problem of conflicts of

interest by offsetting the interested group with

other groups. In view of the need for the technical expertise of the molecular biologists, this

approach seems reasonable; nevertheless the

matter could probably have been handled more

expeditiously. Although the April 1975 amendment to the RAC charter added experts from

such fields as epidemiology and infectious diseases, the charter did not require plant experts

until September 1976 (shortly after the passage

of the original Guidelines) and occupational

health specialists until December 1978. In addition, while two nontechnical members were added in 1976 (one before and one after passage of

the Guidelines), their number was not increased

until secretary Califano reconstituted the Committee in late 1978.

The present makeup of RAC is fairly diverse.

As of September 1980, nine of its members specialized in molecular biology or related fields,

seven were from other scientific disciplines,

and eight were from the areas of law, public

policy, the environment, and public or occupational health.30 Moreover, since December 1978,

representatives of the interested Federal agencies have been sitting as nonvoting members. In

January 1981, an expert on fermentation was

added.

‘N. Wade, “Recombinant DNA: NIH Sets Strict Rules to Launch

New Technology,” 190 Science 1175,1179, 1975.

zsDr. Elizabeth Kutter, a member of RAC at that time, peI’SOnal

communication, Sept. 11, 1980.

sODr. Bernard Talbot, Special Assistant to the Director, NIH, personal communication, Sept. 18, 1980.
Ch. 11-Regulation of Genetic Engineering ● 223

One conflict of interest not solved by expanding the diversity of the RAC’s membership is institutional in nature. NIH, the agency having primary responsibility for developing and administering the Guidelines, views its mission as one of

promoting biomedical research. Although the

Guidelines are not regulations, they contain

many of the elements of regulations. They set

standards, offer a limited means to monitor for

compliance, and provide for enforcement, at

least for institutions receiving NIH grants to do

rDNA work; thus, they may be considered

quasi-regulatory. Regulation is not only foreign

but antithetical to NIH’s mission. The current

Director stated publicly at the June 1980 RAC

meeting that the role of NIH is not one of a

regulator, a role that must be avoided. Under

these circumstances, perhaps another agency,

or another part of DHHS, might be more appropriate for overseeing the Guidelines, since the

attitudes and priorities of promoters are usually

quite different from that of regulators.

If RAC has always been essentially a technical

advisory body, who then has made the value decisions concerning the acceptability of the risks

presented by rDNA and the means for dealing

with them? The final decisionmaker has been

the Director of NIH, with the notable exception

in the case of the 1978 Guidelines, which contained the significant procedural revisions

needed to meet Secretary Califano’s approval.31

The Director did have access to diverse points

of view through the Director’s Advisory Committee meetings and the public hearings held

before the 1978 Guidelines. (See app. III-A.) In

addition, major actions were always accompanied by a statement discussing the relevant

issues and explaining the basis for the decisions;

after the 1978 revisions, major actions had to be

proposed for public comment before decisions

were made. In theory, it may have been preferable for the public to have been substantially involved in the actual formation of the original

Guidelines rather than simply to have reacted to

a finished product. However, this probably

would have slowed the process at a time when

the, strong desire of the molecular biologists to

3’Califano, op. cit.

●

use the rDNA techniques could have threatened

the notion of self-regulation. Today, there appears to be reasonable opportunity for public

input through the process of commenting on

proposed actions.

Conclusion

The Guidelines are the result of an extraordinary, conscientious effort by a combination of

scientists, the public, and the Federal Government, all operating in an unfamiliar realm. They

appear to be a reasonable solution to the problem of how to minimize the risks to health and

the environment posed by rDNA research in an

academic setting, while permitting as much of

that research as possible to proceed. They do

not in any way deal with other molecular genetic techniques or with the long-term social or

philosophical issues that may be associated with

genetic engineering.

The Guidelines have been an evolving document. As more has been learned about rDNA

and molecular genetics, containment levels

have been significantly lowered. Also, the degree of Federal oversight has been substantially

lessened. Under the November 1980 Guidelines,

virtually all responsibility for monitoring compliance is placed on the IBCs. NIH’s role will involve primarily: 1) continuing interpretation of

the Guidelines, 2) certifying new host-vector

systems, 3) serving as a clearinghouse of information, 4) continuing risk assessment experiments, and 5) coordinating Federal and local activities.

The most significant short-term limitation of

the Guidelines is the way they deal with commercial applications and products of rDNA techniques. Although large-scale containment levels

and related administrative procedures exist,

there are several reasons for questioning the effectiveness of the voluntary compliance concept. The most serious problem has been the

lack of expertise in fermentation technology on

RAC. In addition, since the Guidelines are not

legally binding upon industry, the NIH lacks enforcement authority, although there has been

no evidence of industrial noncompliance. Finally, because of its role as a promoter of bio-
224 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

medical research, NIH cannot be expected to act procedural safeguards designed to accommoaggressively to fill this regulatory void. date social values and to limit conflicts of inAs a model for societal decisionmaking on terest. The only major criticism is that procedural safeguards and public input were not sig- technological risks, the system created by the

Guidelines could serve as a valuable precedent. nificant factors when the rDNA problem was

first addressed. It does a reasonable job of combining substantive scientific evaluation of technical issues with

other means of regulation

There are three other means available for

regulating molecular genetic techniques and

their products—current Federal statutes, tort

law and. workmen’s compensation, and State

and local laws. These all may be used to remedy

some of the limitations of the Guidelines.

Federal statutes

The question of whether existing Federal

statutes provide adequate regulatory authority

first arose with respect to rDNA research. In

March 1977, the Interagency Committee concluded that while a number of statutes* could

provide authority to regulate specific phases of

work with rDNA, no single one or combination

would clearly reach all rDNA research to the extent deemed necessary by the Committee. Furthermore, while some could be broadly interpreted, the Committee believed that regulatory

action taken on the basis of those interpretations would be subject to legal challenge.32 This

was the basis for their conclusion that specific

legislation was needed and was one of the reasons behind the legislative effort discussed in

app. III-A.

With respect to commercial uses and products of rDNA and other genetic techniques, a

much more certain basis for regulation exists.

Many of the Federal environmental, product

safety, and public health laws are directed

toward industrial processes and products. To a

“1’he Committee concentrated on the following statutes: 1) the

Occupational %fety and Health Act (29 IJ.S.C. S651 et. seq.); 2) the

Toxic Sul]stiin~es Control Act (15 IJ.S.(:, $2601 et. seq.); 3) the Hazardous Miiterials ‘1’ransportation Act (49 LJ. S.C. ~ 1801 et, seq.); and

4) sec. 361 of the Public Health Service Act (42 LJ.S.C. $264).

az/nterjm Re/lort Of the Federal Interagency CQnlmittee on Recombinant DNA Research: Suggested Elemenfsfor f.e.gislation, op. cit.

large extent, the genetic technologies will produce chemicals, foods, and drugs—as well as

pollutant byproducts—that will clearly come

within the scope of these laws. ’However, there

may be limitations in these laws and questions

of their interpretation that may arise with respect to the manufacturing process, which

employs large quantities of organisms, and

when there is an intentional release of microorganisms into the environment—e.g., for cleaning up pollution. For a list of pertinent laws, see

table 36.)

The Federal Food, Drug, and Cosmetic Act

(FEDCA) and section 351 of the Public Health

Service Act (42 U.S.C. 262) give FDA authority

over foods, drugs, biological products (such as

vaccines), medical devices, and veterinary medicines. This authority will also apply to those

products when they are made by genetic engiTable 36.—Statutes That Will Be Most Applicable

to Commercial Genetic Engineering

1. Federal Food, Drug, and Cosmetic Act (21 U.S.C. $301

et. seq.)

2. Occupational Safety and Health Act (29 U.S.C. $651 et.

seq.)

3. Toxic Substances Control Act (15 U.S.C. $2601 et.

seq.)

4. Marine Protection, Research, and Sanctuaries Act (33

U.S.C. $1401 et. seq.)

5. Federal Water Pollution Control Act, as amended by

the Clean Water Act of 1977 (33 U.S.C. $1251 et. seq.)

6. The Clean Air Act (42 U.S.C. $7401 et. seq.)

7. Hazardous Materials Transportation Act (49 U.S.C.

$1801 et. seq.)

8. Solid Waste Disposal Act, as amended by the

Resource Conservation and Recovery Act of 1976 (42

U.S.C. $6901 et. seq.)

9. Public Health Service Act (42 U.S.C. $301 et. seq.)

10. Federal Insecticide, Fungicide, and Rodenticide Act (7

U.S.C. $136 et. seq.)

SOURCE: Office of Technology Assessment.
Ch. 11-Regulation of Genetic Engineering . 225

neering methods. However, interpretive questions arising out of the unique nature of the

technologies—such as the type of data necessary to show the safety and efficacy of a new

drug produced by rDNA techniques–will have

to be resolved by the administrative process on

a case-by-case basis.

FDA has not published any statements of official policy toward products made by genetic

engineering. Since it has different statutory authority for different types of products, it is likely that regulation will be on a product-by-product basis through the appropriate FDA bureau.

Substances produced by genetic engineering

will generally be treated as analogous products

produced by conventional techniques with respect to standards for chemistry, pharmacology, and clinical protocols; however, quality controls may have to be modified to assure continuous control of product purity and identity. In

addition, for the time being, the Bureau of

Drugs and the Bureau of Biologics will require a

new Notice of Claimed Investigational Exemption for a New Drug and a new New Drug Application for products made by rDNA technology,

even if identity with the natural substance or

with a previously approved drug is shown. This

policy is based on the position that drugs or

biologics made by rDNA techniques have not

become generally recognized by experts as safe

and effective and therefore meet the statutory

definition of a “new drug. ’’33*

FFDCA also permits regulation of drug, food,

and device manufacturing. Certain FDA regulations, called Good Manufacturing Practices, are

designed to assure the quality of these products.

FDA may have to revise these to accommodate

genetic technologies; it has the authority to do

so. It probably does not have the authority to

use these regulations to address any risks to

workers, the public, or the environment, since

FFDCA is designed to protect the consumer of

the regulated product.

ssMinUteS of the Industrial Practices Subcommittee of the Federal Interagency Advisory Committee on Recombinant DNA Research, Dec. 16, 1980, p. 3.

*Sec. 201(p) of the FFLX:A (21 IJ.S.(:. $321(p)) defines a new drLIg

as “any drug . . . the composition d’ which is such that such drLJg

is not .gen(?rtil]y re(mgnized, i]n](~[l~ t?xperts (Iuiilitied by scientific

tl’ilillill~ and experience . . iIS Silf[? illld Pt”t”(?d i~’e . .”

The statute most applicable to worker health

and safety is the Occupational Safety and Health

Act, which grants the Secretary of Labor broad

power to require employers to provide a safe

workplace for their employees. This power includes the ability to require an employer to

modify work practices and to install control

technology. The statute creates a general duty

on employers to furnish their employees with a

workplace “free from recognized hazards that

are causing or are likely to cause death or serious physical harm, ” and it requires employers

to comply with occupational safety and health

standards set by the Secretary of Labor. According to a recent Supreme Court case, a standard

may be promulgated only on a determination

that it is “reasonably necessary and appropriate

to remedy a significant risk of material health

impairment. ”34 Because these fairly stringent requirements limit the Act’s applicability to

recognized hazards or significant risks, the

statute could not be used to control manufacturing where the genetic techniques presented only hypothetical risks. However, it should be applicable to large-scale processes using known

human toxins, pathogens, or their DNA.

The Secretary of Labor is also directed to account for the “urgency of the need” in establishing regulatory priorities. How the Department of Labor will view genetic technologies within its scale of priorities remains to be

seen. NIOSH, the research organization created

by this statute, has been studying rDNA production methods to determine what risks, if any,

are being faced by workers. It has conducted

fact-finding inspections of several manufacturers, and it is planning a joint project with

EPA to assess the adequacy of current control

technology. In addition, a group established by

the Center for Disease Control (CDC) together

with NIOSH will be making recommendations

on: 1) the medical surveillance of potentially exposed workers, 2) the central collection and

analysis of medical data for epidemiological purposes, and 3) the establishment of an emergency

response team.35

s4/ndustria/ Union Department, AFL-C1O v. American Rtrolellm

Institute, 100 S. Ct. 2844,2863, 1980.

3SMinute~ of the [ndustria] practices Subcommittee of the Federal

Interagency Advisory Committee on Recombinant DNA Research,

Dec. 16, 1980, op. cit., p, 6.
226 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

The Toxic Substances Control Act (TSCA) was

intended by Congress to fill in the gaps in the

other environmental laws. It authorizes EPA to

acquire information on “chemical substances” in

order to identify and evaluate potential hazards

and then to regulate the production, use, distribution, and disposal of those substances.

A "chemical substance” is defined under section 3(2) of this Act as “any organic or inorganic

substance of a particular molecular identity, ” including “any combination of such substances occurring in whole or in part as a result of a chemical reaction or occurring in nature.”* This

would include DNA molecules; however, it is

unclear if the definition would encompass genetically engineered organisms. In promulgating

its Inventory Reporting Regulations under

TSCA on December 23, 1977, EPA took the following position in response to a comment that

commercial biological preparations such as

yeasts, bacteria, and fungi should not be considered chemical substances: 36

The Administrator disagrees with this comment . . . . This definition [of chemical substance] does not exclude life forms which may

be manufactured for commercial purposes and

nothing in the legislative history would suggest

otherwise.

However, in a December 9, 1977, letter responding to a Senate inquiry, EPA Administrator Douglas M. Costle stated:37

[A]lthougkt there is a general consensus that recombinant DNA molecules are “chemical substances” within the meaning of section 3 of

TSCA, it is not at all clear whether a host organism containing recombined DNA molecules

fits—or was intended to fit—that definition . . . .

If such organisms are subject to TSCA on the

grounds that they are a “combination of . . .

substances occurring in whole or in part as a

result of a chemical reaction,” the Agency might

logically have to include all living things in the

definition of “chemical substance’’—an inter-

“Substances subject solely to FFDCA or the Federal Insecticide,

Fungicide, and Rodenticide Act are excluded from this definition.

3642 F. R., 64572, 64584, Dec. 23, 1977.

Sletter to Adlai E. Stevenson, Chairman, Subcommittee on Science, Technology, and Space, U.S. Senate Committee on Commerce, Science, and Transportation, in Overs@t Report, lhxombinant DIVA Reseamh and Its Applications, 95th Cong., 2d sess., August 1978, p.88.

pretation which I am confident the Congress

neither contemplated nor intended.

If EPA were to take the broader interpretation, and if that were to survive any legal challenge, TSCA would have great potential for regulating commercial genetic engineering by regulating the organisms. Under section 4 of this

Act, EPA can adopt rules requiring the testing of

chemical substances that “may present an unreasonable risk”* to health or the environment

when existing data are insufficient to make a

determination. Under section 5, the manufacturer of a new chemical substance is required to notify EPA 90 days before beginning

production and to submit any test data available

on the chemical’s health or environmental effects. If EPA decides that the data are insufficient for evaluating the chemical’s effects and

that it “may present an unreasonable risk” or

will be produced in substantial quantities, the

chemical substance’s manufacture or use can be

restricted or prohibited. Under section 6, EPA

can prohibit or regulate the manufacture or use

of any chemical substance that “presents, or will

present an unreasonable risk of injury to health

or the environment. ”

As with the Occupational Safety and Health

Act, the scientific evidence probably does not

support a finding that most genetically engineered molecules or organisms present an unreasonable risk. On the other hand, the standard in section 5—may present an unreasonable

risk—and the requirement for a premanufacturing notice would permit EPA to evaluate

cases where genetically engineered micro-organisms were proposed to be released into the

environment.

Several other environmental statutes will apply, mainly with respect to pollutants, wastes,

or hazardous materials.** The Marine Protec-

*The term “unreasonable risk” is not defined in the statute.

However, the legislative history indicates that its determination involves balancing the probability that harm will occur and the

magnitude and severity of that harm, against the effect of the proposed regulatory action and the availability to society of the benefits of the substance.sa

3JJH. Rept. W. 1341, ~dth [:ong., 2d sess. 1976, pp. 13-15.

* *Two consumer protection statutes were considered but were

determined to be virtually inapplicable These were: the Federal

Hazardous Sul}stances Act (15 11. S.(;. 31261 et. seq.); and the Consumer Product Safet.v Act [15 IJ. S.C. S2051 et. seq.).
Ch. 11-Regulation of Genetic Engineering . 227

tion, Research, and Sanctuaries Act prohibits

ocean dumping without an EPA permit of any

material that would “unreasonably degrade or

endanger human health, welfare, or amenities,

or the marine environment, ecological systems,

or economic potentialities. ”39 “Material” is defined as “matter of any kind or description, including. . . biological and laboratory waste

. . . and industrial . . . and other waste. ”40 The

Federal Water Pollution Control Act regulates

the discharge of pollutants (which include biological materials) into U.S. waters, and the Solid

Waste Disposal Act regulates hazardous wastes.

The Clean Air Act regulates the discharge of air

pollutants, which includes biological materials.

Especially applicable is section 112 (42 U.S.C. 

7412), which allows EPA to set emission standards for hazardous air pollutants—those for

which standards have not been set under other

sections of the Act and which “may reasonably

be anticipated to result in an increase in mortality or an increase in serious irreversible, or incapacitating reversible, illness. ” The Hazardous

Materials Transportation Act covers the interstate transportation of dangerous articles, including etiologic (disease-causing) agents. The

Secretary of Transportation may designate as

hazardous any material that he finds “may pose

an unreasonable risk to health and safety or

property” when transported in commerce in a

particular quantity and form.41

Section 361 of the Public Health Service Act

(42 U.S.C. $264) authorizes the Secretary of

HEW (now DHHS) to “. . . make and enforce

such regulations as in his judgment are necessary to prevent the introduction, transmission,

or spread of communicable diseases . . . .“ Because of the broad discretion given to the Secretary, it has been argued that this section provides sufficient authority to control all rDNA activities. * Others have argued that its purpose is

to protect only human health; for regulations to

be valid, there would have to be a supportable

finding of a connection between rDNA and

3933 U.s.c. $1412.

to 33 U.S.CO ~ 1402(c).

4149 U.S.C. ~ 1803.

“On Nov. 11, 1976, the Natural Resources Defense Council and

the Environmental Defense Fund petitioned the Secretary of HEW

to promulgate regulations concerning rDNA under this Act.

human disease. In any event, HEW declined to

promulgate any regulations.

The following conclusions can therefore be

made on the applicability of existing statutes.

First, the products of genetic technologies—

such as drugs, chemicals, pesticides,** and

foods—would clearly be covered by statutes

already covering these generic categories of

materials. Second, uncertainty exists for regulating either production methods using engineered micro-organisms or their intentional

release into the environment, when risk has not

been clearly demonstrated. Third, the regulatory agencies have begun to study the situation but, have not promulgated specific regulations. Fourth, since regulation will be dispersed throughout several agencies, there may

be conflicting interpretations unless active efforts are made by the Federal Interagency Committee to develop a comprehensive, coordinated

approach.

Tort law and workmen’s compensation

Statutes and regulations are usually directed

at preventing certain types of conduct. While

tort law strives for the same goal, its primary

purpose is to compensate injuries. (A tort is a

civil wrong, other than breach of contract, for

which a court awards damages or other relief. )

By its nature, tort law is quite flexible, since it

has been developed primarily by the courts on a

case-by-case basis. Its basic principles can easily

be applied to cases where injuries have been

caused by a genetically engineered organism,

product, or process. It therefore can be applied

to cases involving genetic technologies as a

means of compensating injuries and as an incentive for safety-conscious conduct. The most applicable concepts of tort law are negligence and

strict liability. (A related body of law—workmen’s compensation—is also pertinent. )

Negligence is defined as conduct (an act or an

omission) that involves an unreasonable risk of

harm to another person. For the injured party

to be compensated, he must prove in court that:

1) the defendant’s conduct was negligent, 2) the

● ● Pesticides iire sut}ject to the ti’ederal Insecticide Fungicide,

iind Rodt)tlt icidr A(Y, 7 L r. S.(; . ~ 136 et. S(>(l..
228 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

defendant’s actions in fact caused the injury,

and 3) the injury was not one for which compensation should be denied or limited because

of overriding policy reasons.

Because of the newness of genetic technology, legal standards of conduct (e.g., what constitutes unreasonable risk) have not been articulated by the courts. If a case were to arise, a

court would undoubtedly look first to the

Guidelines. Even if a technique other than rDNA

were involved, they would provide a general

conceptual framework for good laboratory and

industrial techniques. Other sources for standards of conduct include: 1) CDC’s guidelines for

working with hazardous agents; 2) specific Federal laws or regulations, such as those under the

Public Health Service Act covering the interstate transportation of biologic products and

etiologic agents; and 3) industrial or professional codes or customary practices, such as

generally accepted containment practices in the

pharmaceutical industry or in a microbiology

laboratory. Compliance with these standards,

however, does not foreclose a finding of negligence, since the courts make the ultimate judgment of what constitutes proper conduct. In

several cases, courts have decided that an entire

industry or profession has lagged behind the

level of safe practices demanded by society. *

Conversely, noncompliance with existing standards almost surely will result in a finding of

negligence, if the other elements are also present.

Causation may be difficult to prove in a case

involving a genetically engineered product or

organism. In the case of injury caused by a pathogenic micro-organism—e.g., it may be difficult

to isolate and identify the micro-organism and

virtually impossible to trace its origin, especially

if it had only established a transitory ecological

niche. In addition, it might be difficult to

reconstruct the original situation to determine

if the micro-organism simply escaped despite

‘For example, see: The T. J. Hooper, 60 F. 2d 737 (2d Cir. 1932),

concerning tugboats; and He/hhg v. Carey, 519 P. 2d 981 (1974),

where the court held that the general practice among ophthalmologists of not performing glaucoma tests on asymptomatic patients

under 40 [because they had only a one in 25,000 chance of having

the disease) would not prevent a finding of negligence when such

a patient developed the disease.

precautions or if culpable human action was involved. On the other hand, if a micro-organism

or toxin is identified, it may be so unique

because of its engineering that it can be readily

associated with a company known to produce it

or with a scientist known to be working with

it. * *

The law recognizes that not every negligent

act or omission that causes harm should result

in liability and compensation—e.g., the concept

of “foreseeable” harm serves to limit a defendant’s liability. The underlying social policy

is that the defendant should not be liable for injuries so random or unlikely as to be not reasonably foreseeable. This determination is made

by the court. In the case of a genetically engineered organism, extensive harm would probably be foreseeable because of the organism’s

ability to reproduce; how that harm could occur

might not be foreseeable.

Unlike negligence, strict liability does not require a finding that the defendant breached

some duty of care owed to the injured person;

the fact that the injury was caused by the defendant’s conduct is enough to impose liability

regardless of how carefully the activity was

done. For this doctrine to apply, the activity

must be characterized as “abnormally dangerous.” To determine this, a court would look

at the following six factors, no one of which is

determinative:42

1. existence of a high risk of harm,

2. great gravity of the harm if it occurs,

3. inability to eliminate the risk by exercising

reasonable care,

● “If several companies were working with the micro-organism,

it could be impossible to prove which company produced the particular ones that caused the harm. A recent California Supreme

Court case, Sindell v. Abbott Laboratories, 26 CaL3d 588, 1980,

could provide a way around this problem if the new theory of

liability that it establishes becomes widely accepted by courts in

other jurisdictions. The Court ruled that women whose mothers

hiid taken diethy]stilbest rol, a drug that allegedly caused cancer in

their daughters, could proceed to trial against manufacturers of

the drug, even though most of the plaintiffs would not be able to

show which particular manufacturers produced the drug. The

Court said that when the defendant manufacturers had a substantial share of the product market, liability, if found, would be apportioned among the defendants on the basis of their market

share. A particular defendant could escape liability only by

proving it could not have made the drug.

dz~es.tatement &cond) of Torts $520 (1976).
Ch. 11—Regulation of Genetic Engineering . 229

4. extent to which the activity is not common,

5. inappropriateness of the activity to the

place where it is done, and

6. the activity’s value to the community.

Given the current consensus about the risks

of genetic techniques, it would be difficult to

argue that the doctrine of strict liability should

apply. However, in the extremely unlikely event

that a serious, widespread injury does occur,

that alone would probably support a court’s determination that the activity was abnormally

dangerous, regardless of its probability. In such

cases, the courts have generally relied on the

principle of “enterprise liability ’’–that those engaged in an enterprise should bear its costs, including the costs of injuries to others.43

For either negligence or strict liability, the

person causing the harm is liable. Under the

legal principle of respondent superior, liability is

also imputed from the original actor to people

or entities who have a special relationship with

him—e.g., employers. Thus, a corporation can

be liable for the torts of its scientists or production workers. Similarly, a university, an IBC, a

Biological Safety Officer, and a PI would probably be liable for the torts of scientists and students under their direction.

Another body of law designed to compensate

injuries deserves brief mention. Workmen’s

compensation is a statutory scheme adopted by

the States and—for specific occupations or circumstances—by the Federal Government to

compensate injuries without a need for showing

fault. The employee need only show that the injury was job-related. He is then compensated by

the employer or the employer’s insurance company. It would clearly apply to genetic engineering.

Tort law and workmen’s compensation will

be available to compensate any injuries resulting from the use of molecular genetic techniques, especially from their commercial application. Tort law may also indirectly prevent

potentially hazardous actions, although the de43R. DWOrkln, “giocatastrophe and the Law: Legal Aspects of Recombinant DNA Research, ” in The Recombinant DNA Debate,

Jackson and Stitch (eds.) (Englewood Cliffs, N. J.: Prentice-Hal], Inc.

1979), pp. 219, 223.

terrent effect of compensation is less efficient

than direct regulation—e.g., the threat of lawsuits will not necessarily discourage high-risk

activities where problems of proof make recovery unlikely, where the harm may be small

and widespread (as with mild illness suffered by

a large number of people), or where profits are

less than the cost of prevention but greater than

expected damage awards and legal costs.

Tort law has two other limitations. First, tort

litigation involves high costs to the plaintiff, and

indirectly to society. Second, it cannot adequately compensate the victims of a catastrophic situation where liability would bankrupt the

defendant.

State and local law

Under the 10th amendment to the Constitution, all powers not delegated to the Federal

Government are reserved for the States or the

people. One of those is the power of the States

and municipalities to protect the health, safety,

and welfare of their citizens. Thus, they can

regulate genetic engineering.

The reasons espoused in favor of local regulation are based on the traditional concept of local

autonomy; those most likely to suffer any

adverse affects of genetic engineering should

control it. Also, local and State governments are

usually more accessible to public input than the

Federal Government. Consequently, judgments

on the acceptability of the risks will more

precisely reflect the will of the segment of the

public most directly affected.

A number of arguments have been made

against local as opposed to Federal regulation.

The primary one is that regulation by States and

communities would give rise to a random patchwork of confusing and conflicting controls. In

addition, States and especially localities may not

have the same access as the Federal Government to the expertise that should be used in the

formulation of rational controls. Finally, any

risks associated with rDNA or other techniques

are not limited by geographic boundaries;

therefore, they ought to be dealt with nationally. The above arguments reflect the position

that regulation of genetic technologies is a na76-565 0 - 81 - 16
230 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

tional issue that can be handled most effectively

at the Federal level.

A few jurisdictions have used their authority

in the case of rDNA, * The most comprehensive

regulation was created by the States of Mary-

● Cambridge, Mass., established a citizens’ study group that rec.

ommended that researchers be subject to some additional re.

striations beyond those of the Guidelines. These were embodied in

an ordinance passed by the City Council on Feb. 7, 1977. Serkeley,

Calif., passed an ordinance requiring private research to conform

to the Guidelines. Similar ordinances or resolutions were passed

by Princeton, N.J., Amherst, Mass., and Emeryville, Calif.

land and New York.4445 Currently, there is little,

if any, effort on the State or local level to pass

laws or ordinances covering rDNA or similar

genetic techniques, and there is little activity

under the existing laws.

4aAnnotated Code of Maryland, art. 43 ~~ 898-910 (SUpp. 19781.

45Mc~jnney’s C“onsoljdated Laws of fVew York, Public Health ~w,

art. 32-A S$3220-3223 (supp. 1980)

Conclusion

The initial question with respect to regulating

genetic engineering is how to define the scope

of the problem. This will depend largely on

what groups are involved in that process and

how they view the nature, magnitude, and

acceptability of the risks. Similarly, the means

of addressing the problem will be determined

by how it is defined and who is involved in the

actual decisionmaking process. For these reasons, it is important that regulatory mechanisms

combine scientific expertise with procedures to

accommodate the values of those bearing the

risk so that society may have confidence in

those mechanisms.

Currently, genetic techniques and their products are regulated by a combination of the

Guidelines, Federal statutes protecting health

and the environment, some State or local laws,

and the judicially created law of torts, which is

available to compensate injuries after they occur. In most cases, this system appears adequate

to deal with the risks to health and the environment. However, there is some concern regarding commercial applications for the following

reasons: 1) the voluntary applicability of the

Guidelines to industry, 2) RAC’s insufficient expertise in fermentation technology, 3) the potential interpretive problems in applying existing law to the workplace and to situations

where micro-organisms are intentionally released into the enviornment, and 4) the absence

of a definitive regulatory posture by the

agencies.

Issue and Options

 ISSUE: How could Congress address the

risks associated with genetic engineering?

A number of options are available, ranging

from deregulation through comprehensive new

regulation. An underlying issue for most of

these options is: What are the constitutional

constraints placed on congressional regulation

of molecular genetic techniques, particularly

when they are used in research? (This is discussed in app. III-B.)

OPTIONS:

A: Congress could maintain the status quo by letting NIH and the regulatory agencies set the

Federal policy.

This option requires Congress to determine

that legislation to remedy the limitations in current Federal oversight would result in unnecessary and burdensome regulation. No known

harm to health or the environment has occurred under the current system, and the agencies generally have significant legal authority
Ch. 11-Regulation of Genetic Engineering . 231

and expertise that should permit them to adapt

to most new problems posed by genetic engineering. The agencies have been consulting

with each other through the Interagency Committee, and the three agencies that will play the

most important role in regulating large-scale

commercial activities—FDA, OSHA, and EPA—

have been studying the situation.

The disadvantages of this option are the lack

of a centralized, uniform Federal response to

the problem, and the possibility that risks

associated with commercial applications will not

be adequately addressed. Certain applications,

such as the use of micro-organisms for oil recovery are not unequivocally regulated by current statutes; broad interpretations of statutory

language in order to reach these situations may

be overturned in court. Conflicting or redundant regulations of different agencies would

result in unnecessary burdens on those regulated. In addition, some commercial activity is

now at the pilot plant stage, but the responsible

agencies have yet to establish official policy and

to devise a coordinated plan of action.

B: Congress could require that the Federal Interagency Advisory Committee on Recombinant

DNA Research prepare a comprehensive report on its members’ collective authority to

regulate rDNA and their regulatory intentions.

The Industrial Practices Subcommittee of this

Committee has been studying agency authority

over commercial rDNA activities. Presently,

there is little official guidance on regulatory requirements for companies that may soon market products made by rDNA methods. -e.g.,

companies are building fermentation plants

without knowing what design or other requirements OSHA may mandate for worker safety.

As was stated by former OSHA head, Dr. Eula

Bingham, it will take at least 2 years for OSHA to

set standards, if the current NIOSH study shows

a need for them.46

A congressionally mandated report would

assure full consideration of these issues by the

agencies and expedite the process. It could in46[,etter from Dr, ~llla Bingham, Assistant %?cretary for ~ccupational Safety and Health, to Dr. Donald Fredrickson, Director, NIH,

Sept. 24, 1980.

elude the following: 1) a section prepared by

each agency that assesses its statutory authority

and articulates what activities and products will

be considered to come within its jurisdiction, 2)

a summary section that evaluates the adequacy

of existing Federal statutes and regulations as a

whole with respect to commercial genetic engineering, and 3) a section proposing any specific legislation considered to be necessary.

The principal disadvantages of this option are

that it maybe unnecessary and impractical. The

agencies are studying the situation, which must

be done before they can act. Also, it is often

easier and more efficient to act on each case as

it arises, rather than on a hypothetical basis

before the fact.

C: Congress could require Federal monitoring of

all rDNA activity for a limited number of

years.

This option represents a “wait and see” position by Congress and the middle ground between the status quo and full regulation. It recognizes and balances the following factors: 1)

the absence of demonstrated harm to human

health or the environment from genetic engineering; 2) the continuing concern that genetic engineering presents risks; 3) the lack of sufficient knowledge from which to make a final

judgment; 4) the existence of an oversight mechanism that seems to be working well, but that

has clear limitations with respect to commercial

activities; 5) the virtual abolition of Federal

monitoring of rDNA activities by the recent

amendments to the Guidelines; and 6) the expected increase in commercial genetic engineering activities.

Monitoring involves the collection and evaluation of information about an activity in order

to know what is occurring, to determine the

need for other action, and to be able to act if

necessary. More specifically, this option would

provide a data base that could be used for: 1) determining the effectiveness of voluntary compliance with the Guidelines by industry and mandatory compliance by Federal grantees, 2) determining the quality and consistency of IBC decisions and other actions, 3) continuing a formal

risk assessment program, 4) identifying vague
232 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

or conflicting provisions of the Guidelines for

revision, S) identifying emerging trends or problems, and 6) tracing any long-term adverse impacts on health or the environment back to

their sources.

The obvious disadvantages of this option are

the increased paperwork and effort by scientists, universities, corporations, and the Federal

Government. Those working with rDNA would

have to gather the required information periodically and prepare reports, which would be

filed by the sponsoring institution with a

designated existing Federal agency. A widerange of information would be required for

each project. The agency would have to process

the reports and take other actions, such as preparing an annual report to Congress, to implement the underlying purposes of this option.

Additional manpower would most likely be

needed by that agency.

A statute implementing this option could include the following elements: 1) periodic collection of information in the form of reports from

all institutions in the United States that sponsor

any work with rDNA, 2) active evaluation of

that information by the collecting agency, 3) annual reports to Congress, and 4) a sunset clause.

Important information would include: 1) the

sponsoring institution’s name; 2) all places

I where it sponsors the research; and 3) a tabular

or other summary that discloses for each project continuing or completed during the reporting period: the culture volume, the source and

identity of the DNA and the host-vector system,

the containment levels, and other information

deemed necessary to effect the purposes of the

act. The statute could also require employers to

institute and report on a worker health surveillance program.

For this option to work, the monitoring agency would have to take an active role in evaluating the data. It should have the authority to

require amendments to the reports when any

part is vague, incomplete, or inconsistent with

another part. It could also be required to notify

the appropriate Federal funding agency of apparent cases of noncompliance with the Guidelines by their grantees. Finally, it should prepare an annual report to Congress on the effectiveness of Federal oversight.

The choice of an agency to administer the

statute would be important. The selection of

NIH would permit the use of an existing administrative structure and body of expertise and experience. On the other hand, one of the regulatory agencies may take a more active monitoring role and be more experienced with

handling proprietary information.

This approach is similar to a bill introduced in

the 96th Congress, S. 2234, but broader in

scope. The latter covered only institutions not

funded by NIH, and did not contain provisions

for requiring amendments to the reports or for

notifying other agencies of possible noncompliance. The bill was broader in one respect

because it would have required information

about prospective experiments. This provision

had been criticized because of the difficulty of

projecting in advance the course that scientific

inquiry will take. The goals of a monitoring program can be substantially reached by monitoring ongoing and completed work.

D. Congress could make the NIH Guidelines applicable to all rDNA work done in the United

States.

The purpose of this option is to alleviate any

concerns about the effectiveness of voluntary

compliance. RAC itself has gone on record as

supporting mandatory compliance with the

Guidelines by non-NIH funded institutions, including private companies.

This option has the advantages of using an existing oversight mechanism, which would simply be extended to industry and to academic research funded by agencies other than NIH. Specific requirements on technical questions such

as containment levels, host-vector systems, and

laboratory practices would continue to be set by

NIH in order to accommodate new information

expeditiously; the statute would simply codify

the responsibilities and procedures of the current system. There would be few transitional

administrative problems, since the expertise

and experience already exist at NIH. However, it

would be necessary to appoint several experts
Ch. 11-Regulation of Genetic Engineering ● 233

in fermentation and other industrial technologies to RAC if production, as well as research, is

to be adequately covered. In addition, the recommendations for large-scale containment procedures would have to be made part of the

Guidelines.

The major changes would have to be made

with respect to enforcement. Present penalties

for noncompliance—suspension or termination

of research funds—are obviously inapplicable to

industry. In addition, procedures for monitoring compliance could be strengthened. Some of

the elements of option C could be used. An

added or alternative approach would be to inspect facilities.

The main disadvantage of this option is that

NIH is not a regulatory agency. Since NIH has

traditionally viewed its mission as promoting

biomedical research, it would have a conflict of

interest between regulation and promotion.

One of the regulatory agencies could be given

the authority to enforce the Guidelines and to

adopt changes therein. NIH could then continue

in a scientific advisory role.

E. Congress could require an environmental impact statement and agency approval before

any genetically engineered organism is intentionally released into the environment.

There have been numerous cases where an

animal or plant species has been introduced into

a new environment and has spread in an uncontrolled and undesirable fashion. One of the

early fears about rDNA was that a new pathogenic or otherwise undesirable micro-organism could establish an environmental niche.

Yet in pollution control, mineral leaching, and

enhanced oil recovery, it might be desirable to

release large numbers of engineered micro-organisms into the environment.

The Guidelines currently prohibit deliberate

release of any organism containing rDNA without approval by the Director of NIH on advice of

RAC. The obvious disadvantage of this prohibition is that it lacks the force of law. The release

of such an organism without NIH approval

would be a prima facie case of negligence, if the

organism caused harm. However, it may be

more desirable social policy to attempt to prevent this type of harm through regulation

rather than to compensate for injuries through

lawsuits. Another possible disadvantage of the

present system is that approval maybe granted

on a finding that the release would present “no

significant risk to health or the environment;” a

tougher or more specific standard than this may

be desirable.

A required study of the possible consequences following the release of a genetically

engineered organism, especially a micro-organism, would be an important step in ensuring

safety. This option could be implemented by requiring those proposing to release the organism

to file an impact statement with an agency such

as NIH or EPA, which would then grant or deny

permission to release the organism. A disadvantage of this option is that companies and individuals might be discouraged from developing

useful organisms if this process became too

burdensome and costly.

F. Congress could pass legislation regulating all

types and phases of genetic engineering from

research through commercial production.

The main advantage of this option would be

to deal comprehensively and directly with the

risks of novel molecular genetic techniques,

rather than relying on the current patchwork

system. A specific statute would eliminate the

uncertainties over the extent to which present

law covers particular applications of genetic engineering, such as pollution control, and any

concerns about the effectiveness of voluntary

compliance with the Guidelines.

Other molecular genetic techniques, while

not as widely used and effective as rDNA, raise

similar concerns. Of the current techniques, cell

fusion is the prime candidate for being treated

like rDNA in any regulatory framework. It permits the recombination of chromosomes of

species that do not recombine naturally, and it

may permit the DNA of latent viruses in the cells

to recombine into harmful viruses. No risk assessment of this technique has been done, and

no Federal oversight exists.

The principal arguments against this option

are that the current system appears to be working fairly well, and that the limited risks of the
234 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

techniques may not warrant the significantly increased regulatory burden and costs that would

result from such legislation. Congress will have

to decide if that system will remain adequate as

commercial activity grows.

If Congress were to decide on this option, the

legislation could incorporate some or all of options C, D, and E. The present mechanism

created by the Guidelines could be appropriately modified to provide the regulatory framework. The modifications could include a registration and licensing system to provide information on what work was actually being done

and a means for continuous oversight. One

important type of information would be health

and safety statistics gathered by monitoring

workers involved in the production of products

from genetically engineered organisms. Another modification could be a sliding scale of

penalties for violations, ranging from monetary

fines through revocation of operating licenses

to criminal penalties for extreme cases.

It would not be necessary to create a new

agency, which would duplicate some of the responsibilities of existing agencies. Instead, Congress could give these agencies clear regulatory

authority by amending the appropriate statutes.

Designating a lead agency would assure a more

uniform interpretation and application of the

laws.

G. Congress could require NIH to rescind the

Guidelines.

This option requires Congress to determine

that the risks of rDNA techniques are so insignificant that no control or oversight is necessary. Deregulation would have the advantage

of allowing funds and personnel currently involved in implementing the Guidelines at the

Federal and local levels to be used for other purposes. In fiscal year 1980, NIH spent approximately $500,000 in administering the Guidelines; figures are not available for the analogous

cost to academia and industry. Personnel hours

spent have not been estimated. Very few people

work full-time on administering or complying

with the Guidelines. NIH employs only six people full-time for this purpose, and some institutions employ full-time biological safety personnel. However, over 1,000 people nationally

devote some effort to implementing the

Guidelines—members of the IBCs and the scientists conducting the rDNA experiments who

must take necessary steps to comply.

There are several reasons for retaining the

Guidelines. First, sufficient scientific concern

about risks exists for the Guidelines to prohibit

certain experiments and require containment

for others. Second, they are not particularly

burdensome, since an estimated 80 to 85 percent of all experiments can be done at the

lowest containment levels and an estimated 97

percent will not require NIH approval. Third,

NIH will continue to serve an important role in

continuing risk assessments, in evaluating new

host-vector systems, in collecting and dispersing

information, and in interpreting the Guidelines.

Fourth, if the Guidelines were abolished, regulatory activity at the State and local levels could

again become active. Finally, the oversight system has been flexible enough in the past to liberalize restrictions as evidence indicated lower

risk.

H. Congress could consider the need for

regulating work with all hazardous microorganisms and viruses, whether or not they

are genetically engineered.

Micro-organisms carrying rDNA, according to

an increasingly accepted view, represent just a

subset of micro-organisms and viruses, which,

in general, pose risks. CDC has published guidelines for working with hazardous agents such as

polio virus. However, such work is not currently subject to legally enforceable Federal regulations. It was not within the scope of this

study to examine this issue, but it is an emerging

one that Congress may wish to consider.
Chapter 12

Patenting Living Organisms
Chapter 12

Page

A Landmark Decision . . , . , . . . . . . . . , . . . . . . .237

Legal Protection of Inventions, , . . . . . . . . . . , .. 237

Trade Secrets. . . . . . . . . . . . . . . . . . . . . . . . . .237

Patents. . . . . . . . . . . . . , . . . . . . . . , . . . . . . . .238

Living organisms. , . . , . . . . . . . . . . . . . . . . . .239

The Chakrabarty Case. . . . . . . . , . . . . . . . . . ...240

Potential Impacts of the Decision and

Related Policy Issues. . . . . . . . . . . . . ......242

lmpacts on Industry. . . . . . . . . . . . . . . . . . .. .242

The Relationship Between Patents and

Innovation. . . . . . . . . . . . . . . . . . . ......242

The Advantages of Patenting Living

Organisms. . . . . . . . . . , . . . . . . . . . . . . . .243

Patent v. Trade Secret Protection . . . . . . . . .245

Impact of the Court’s Decision on the

Biotechnology Industry . ..............246

Impacts of the Court’s Decision on the Patent

Law and the Patent and Trademark Office ..246

Impact of the Court’s Decision on Academic

Research. . . . . . . . . . . . . . . . . . . . . . ......248

Impacts of the Court’s Decision on Genetic

Diversity and the Food Supply. . . . . . . . . . . .249

The Morality of Patenting Living Organisms. . .250

Private Ownership of Inventions From

Publicly Funded Research. . .............250

Issue and Options . . . . . . . . . . . . . . . . . . . . . . . .252
Chapter 12

Patenting Living Organisms

A landmark decision

In a 5 to 4 decision (Diamond v. Chakrabarty,

June 16, 1980), the Supreme Court ruled that a

manmade mice-organism is patentable under

the current patent statutes. This decision was

alternately hailed as having “assured this country’s technology future”l and denounced as creating “the Brave New World that Aldous Huxley

warned of. ”2 However, the Court clearly stated

that it was undertaking only the narrow task of

determining whether or not Congress, in enacting the patent statutes, had intended a manmade micro-organism to be excluded from patentability solely because it was alive. Moreover,

the opinion invited Congress to overturn the

decision if it disagreed with the Court’s interpretation.

‘Prepared Statement of C,enentech, Inc., cited in “Science May

Patent New Forms of Life, Justices Rule, 5 to 4,” The New York

‘rimes, June 17, 1980, p. 1.

‘Prepared statement of the Peoples’ Business Commission, cited

in “Science May Patent New Forms of Life, Justices Rule, 5 to 4,”

The New York Times, June 17, 1980, p. 1.

Congress may want to reconsider the issue of

whether and to what extent it should specifically provide for or prohibit the patentability of

living organisms. While the judiciary operates

on a case-by-case basis, Congress can consider

all the issues related to patentability at the same

time, gathering all relevant data and taking testimony from the interested parties. The issues

involved go beyond the narrow ones of scientific capabilities and the legal interpretations of

statutory wording. They require broader decisions based on public policy and social values;

Congress has the constitutional authority to

make those decisions for society. It can act to resolve the questions left unanswered by the

Court, overrule the decision, or develop a comprehensive statutory approach, if necessary.

Most importantly, Congress can draw lines; it

can specifically decide which organisms, if any,

should be patentable.

Legal protection of inventions

The inherent “right” of the originator of a

new idea to that idea is generally recognized, at

least to the extent of deserving credit for it

when used by others. At the same time, it is also

believed that worthwhile ideas benefit society

when they are widely available. Similarly, when

an idea is embodied in a tangible form, such as

in a machine or industrial process, the inventor

has the “right” to its exclusive posession and use

simply by keeping it secret. However, if he may

be induced to disclose the invention’s details,

society benefits from the new ideas embodied

therein, since others may build upon the new

knowledge. The legal system has long recognized the competing interests of the inventor

and the public, and has attempted to protect

both. The separate laws covering trade secrets

and patents are the mean by which this is done.

Trade secrets

The body of law governing trade secrets recognizes that harm has been done to one person

if another improperly obtains a trade secret and

then uses it personally or discloses it to others.

A trade secret is anything—device, formula, or

information—which when used in a business

provides an advantage over competitors ignorant of it—e.g., improper acquisition includes

a breach of confidence, a breach of a specific

promise not to disclose, or an outright theft.

Trade secrecy is derived from the common law,

237
238 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

as opposed to being specifically created by

statute; the State courts recognize and protect it

as a form of property. The underlying policy is

one of preventing unfair competition or unjust

benefits. The protection lasts indefinitely. Two

well-known examples of long-time trade secrets

are the formulas for Coca Cola and for Smith

Brothers’ black cough drops; the latter is supposedly over 100 years old.

A company relying on trade secrecy to protect an important invention must take several

steps to effect that protection. These include:

permitting only key personnel to have access,

requiring such people to sign complex contracts

involving limitations on subsequent employment, and monitoring employees and competitors for possible breaches of security. Even

so, there are practical limitations to what can be

done and what can be proved to the satisfaction

of a court. Moreover, independent discovery of

the secret by a competitor is not improper, including the discovery of a secret process by an

examination of the commercially marketed

product. Most importantly, once a trade secret

becomes public through whatever means, it can

never be recaptured. Thus, reliance on trade

secrecy for protecting inventions can be risky.

! Patents

I

I In contrast to the common law development

of trade secrecy, patent law is a creation of Congress. The Federal patent statutes (title 35 of the

United States Code) are derived from article

section 8, of the Constitution, which states:

The Congress shall have Power . . . To promote the Progress of Science and useful Arts, by

securing for limited Times to Authors and Inventors the exclusive Right to their respective

Writings and Discoveries.

I,

This clause grants Congress the power to create a Federal statutory body of law designed to

encourage invention by granting inventors a

lawful monopoly for a limited period of time.

Under the current statutory arrangement,

which is conceptually similar to the first patent

statutes promulgated in 1790, a patent gives the

inventor the right to exclude all others from

making, using, or selling his invention within

the United States without his consent for 17

years. In return, the inventor must make full

public disclosure of his invention. The policy behind the law is twofold. First, by rewarding successful efforts, a patent provides the inventor

and those who support him with the incentive

to risk time and money in research and development. Second, and more importantly, the patent

system encourages public disclosure of technical information, which may otherwise have remained secret, so others may use the knowledge. The inducement in both cases is the potential for economic gain through exploitation

of the limited monopoly. Of course, there are

many reasons why this potential may not be

realized, including the existence of competing

products.

To qualify for patent protection, an invention

must meet three statutory requirements: it

must be capable of being classified as a process,

machine, manufacture, or composition of matter; it must be new, useful, and not obvious; and

it must be disclosed to the public in sufficient

detail to enable a person skilled in the same or

the most closely related area of technology to

construct and operate it. Plants that reproduce

asexually may also be patented, but slightly different criteria are used.

Although the categories in the first requirement are quite broad, they are not unlimited. In

fact, the courts have held such things as scientific principles, mathematical formulas, and

products of nature to be unpatentable on the

grounds that they are only discoveries of preexisting things—not the result of the inventive,

creative action of man, which is what the patent

laws are designed to encourage. This concept

was reaffirmed in the Chakrabarty opinion.

The requirement that an invention be useful,

new, and- not obvious further narrows the

range of patentable inventions. Utility exists if

the invention works and would have some benefit to society; the degree is not important. Novelty signifies that the invention must differ from

the “prior art” (publicly known inventions or

knowledge). Novelty is not considered to exist,

—e.g., if: 1) the applicant for a patent is not

the inventor, 2) the invention was previously
Ch. 12—Patenting Living Organisms ● 239

known or used publicly by others in the United

States, or 3) the invention was previously described in a U.S. or foreign patent or publication. The inability to meet the novelty requirement is another reason why products of nature

are unpatentable. Nonobviousness refers to the

degree of difference between the invention and

the prior art. If the invention would have been

obvious at the time it was made to a person with

ordinary skill in that field of technology, then it

is not patentable. The policy behind the dual

criteria of novelty and nonobviousness is that a

patent should not take from the public something which it already enjoys or potentially

enjoys as an obvious extension of current

knowledge.

The final requirement—for adequate public

disclosure of an invention—is known as the enablement requirement. It is designed to ensure

that the public receives the full ,benefit of the

new knowledge in return for granting a limited

monopoly. As a public document, the patent

must contain a sufficiently detailed description

of the invention so that others in that field of

technology can build and use it. At the end of

this description are the claims, which define the

boundaries of the invention protected by the

patent.

The differences between trade secrets and

patents, therefore, center on the categories of

inventions protected, the term and degree of

protection, and the disclosure required. Only

those inventions meeting the statutory requirements outlined above qualify for patents and

then only for a limited time, whereas anything

giving an advantage over business competitors

qualifies as a trade secret for an unlimited time.

A patent requires full public disclosure, while

trade secrecy requires an explicit and often

costly effort to withhold information. The patent law provides rights of exclusion against

everyone, even subsequent independent inventors, while the trade secrecy law protects only

against wrongful appropriation of the secret.

Any organism that both meets the broad definition of a trade secret and may be lawfully

owned by a private person or entity can be protected by that body of law, including microorganisms, plants, animals, and insects. In addition, plants are covered specifically by two Federal statutes, the Plant Patent Act of 1930 and

the Plant Variety Protection Act of 1970. Furthermore, the Supreme Court has now ruled

that manmade micro-organisms are covered by

the patent statutes. Its determination of congressional intent in the Chakrabarty case was

based significantly on an analysis of the two

plant protection statutes.

Patent protection for plants was not available

until Congress passed the Plant Patent Act of

1930, recognizing that not all plants were products of nature because new varieties could be

created by man. This Act covered new and distinct asexually reproduced varieties other than

tuber-propagated plants or those found in nature. * The requirement for asexual reproduction was based on the belief that sexually

reproduced varieties could not be reproduced

true-to-type and that it would be senseless to try

to protect a variety that would change in the

next generation. To deal with the fact that organisms reproduce, the Act conferred the right

to exclude others from asexually reproducing

the plant or from using or selling any plants so

reproduced. It also liberalized the description

requirement for plants. Because of the impossibility of describing plants with the same degree of specificity as machines, their description

need only be as complete as is “reasonably possible.”

By 1970, plant breeding technology had advanced to where new, stable, and uniform varieties could be sexually reproduced. As a result,

Congress provided patent-like protection to

novel varieties of plants that reproduced sexually by passing the. Plant Variety Protection Act

of 1970. Fungi, bacteria, and first-generation

hybrids were excluded. * * Hybrids have a builtLiving organisms

Although the law for protecting inventions is

usually thought of as applying to inanimate objects, it also applies to certain living organisms.

● Approximately 4,500 plant patents have been issued to date,

most for roses, apples, peaches, and chrysanthemums.

● ● Originally, six vegetables—okra, celery, peppers, tomatoes,

carrots, and cucumbers—were also excluded. On Dec. 22, 1980,

President Carter signed legislation (H.R. 999) amending the Plant

Variety Protection Act to include these vegetables, to extend the

term of protection to 18 years, and to make certain technical

changes.
240 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

in protection, since the breeder can control the

inbred, parental stocks and the same hybrid

cannot be reproduced from hybrid seed.

The 1970 Act, administered by the Office of

Plant Variety Protection within the U.S. Department of Agriculture (USDA), parallels the patent

statutes to a large degree. Certificates of Plant

Variety Protection allow the breeder to exclude

others from selling, offering for sale, reproducing (sexually or asexually), importing, or exporting the protected variety. In addition, others

cannot use it to produce a hybrid or a different

variety for sale. However, saving seed for crop

production and for the use and reproduction of

protected varieties for research is expressly

permitted. The term of protection is 18 years.

The Chakrabarty case

In 1972, Ananda M. Chakrabarty, then a research scientist for the General Electric Co., developed a strain of bacteria that would degrade

four of the major components of crude oil. He

did this by taking plasmids from several different strains, each of which gave the original

strain a natural ability to degrade one of the

crude oil components, and putting them into a

single strain. The new bacterium was designed

to be placed on an oil spill to break down the oil

into harmless products by using it for food, and

then to disappear when the oil was gone. Because anyone could take and reproduce the microbe once it was used, Chakrabarty applied for

a patent on his invention. The U.S. Patent and

Trademark Office granted a patent on the process by which the bacterium was developed and

on a combination of a carrier (such as straw)

and the bacteria. It refused to grant patent protection on the bacterium itself, contending that

living organisms other than plant were not

patentable under existing law. On appeal, the

Court of Customs and Patent Appeals held that

the inventor of a genetically engineered microorganism whose invention otherwise met the

legal requirements for obtaining a patent could

not be denied a patent solely because the invention was alive. The Supreme Court affirmed.

The majority opinion characterized the issue

as follows:3

The question before us in this case is a narrow one of statutory interpretation requiring us

3Diamond v. Chakrabarfy, 100 S. Ct. 2204, 2207 (1980).

‘

(Whoever invents or discovers any new and

useful process, machine, manufacture, or composition of matter, or any new and useful improvement thereof, may obtain a patent therefor, subject to the conditions and requirements

of this title. ”

Specifically, we must determine whether respondent’s micro-organism constitutes a “manufacture” or “composition of matter” within the

meaning of the statute.

After evaluating the words of the statute, the

policy behind the patent laws, and the legislative history of section 101 of the patent

statutes and of the two plant protection Acts,

the Court ruled that Congress had not intended

to distinguish between unpatentable and patentable subject matter on the basis of living versus nonliving, but on the basis of “products of

nature, whether living or not, and human-made

inventions. ”4 Therefore, the majority ruled,

“[t]he patentee has produced a new bacterium

with markedly different characteristics from

any found in nature and one having potential

for significant utility. His discovery is not nature’s handiwork, but his own; accordingly it is

patentable subject matter under $l01.”5 The

majority did not see their decision as extending

the limits of patentability beyond those set by

Congress.

The Court found that, in choosing such expansive terms as “manufacture” and “composition of matter”—words that have been in

every patent statute since 1793—Congress plainly intended the patent laws to have a wide

41bid, p, 2,210.

Slbid, p. 2,208.
Ch. 12—Patenting Living Organisms ● 241

scope. Moreover, when these laws were last remodified in 1952, the congressional committee

reports affirmed the intent of congress that patentable subject matter “include anything under

the sun that is made by man. ”6 The Court

acknowledged that not everything is patentable;

laws of nature, physical phenomena, and

abstract ideas are not.

The Court found the Government’s arguments unpersuasive. Specifically, that passing

the Plant Patent Act of 1930 and the Plant Variety Protection Act of 1970, which excluded bacteria, was evidence of congressional understanding that section 101 did not apply to living

organisms; otherwise; these statutes would

have been unnecessary. In disagreeing, the

Court stated that the 1930 Act was necessary to

overcome the belief that even artificially bred

plants were unpatentable products of nature

and to relax the written description requirement, permitting a description as complete as is

“reasonably possible. ” As for the 1970 Act, the

Court stated that it had been passed to extend

patent-like protection to new sexually reproducing varieties, which, in 1930, were believed to

be incapable of reproducing in a stable, uniform

manner. The 1970 Act’s exclusion of bacteria,

which indicated to the Government that Congress had not intended bacteria to be patentable, was considered insignificant for a number of reasons.

The Government had also argued that Congress could not have intended section 101 to

cover genetically engineered micro-organisms,

since the technology was unforeseen at the

time. The majority responded that the very purpose of the patent law was to encourage new,

unforeseen inventions, which was why section

101 was so broadly worded. Furthermore, as

for the “gruesome parade of horribles”7 that

might possibly be associated with genetic engineering, the Court stated that the denial of a

patent on a micro-organism might slow the scientific work but certainly would not stop it; and

the consideration of such issues involves policy

judgments that the legislative and executive

es, R@, N~,, 1979, 82d (:[]ng, , 2d $j~ss. , P. 5, 19,52; H . R. Rq)t . No.

1923, ml IIong., 2d Sess. j p. 6, 1952, cited in Diam~~~ v.

Chakrabarfy, 100” S. Ct. 2204, 2207 [ 1980).

7Diamond t’. Chakrabarty, 100 S. Ct. 2204, 2211 (1980).

branches of Government, and not the courts,

are competent to make. It further recognized

that Congress could amend section 101 to specifically exclude genetically engineered organisms or could write a statute specifically designed for them.

The dissenting Justices agreed that the issue

was one of statutory interpretation, but interpreted section 101 differently. They saw the

two plant protection Acts as strong evidence of

congressional intent that section 101 not cover

living organisms. In view of this, the dissenters

maintained that the majority opinion was actually extending the scope of the patent laws

beyond the limit set by Congress.

The stated narrowness of the Court’s decision

may limit its impact as precedent in subsequent

cases that raise similar issues, although not necessarily. Certainly, the decision applies to any

genetically engineered micro-organism. It is a

technical distinction without legal significance

that most of the work being done on such organisms involves recombinant DNA (rDNA) techniques, which Chakrabarty did not use. The real

question is whether or not it would permit the

patenting of other genetically engineered organisms, such as plants, animals, and insects.

Any fears that the decision might serve as a

legal precedent for the patenting of human beings in the distant future are totally groundless.

Under our legal system, the ownership of humans is absolutely prohibited by the 13th

amendment to the Constitution.

Although the Chakrabarty case involved a

micro-organism, there is no reason that its rationale could not be applied to other organisms.

In the majority’s view, the crucial test for patentability concerned whether or not the microorganism was manmade. Conceptually, there is

nothing in this test that limits it to microorganisms. The operative distinction is between

humanmade and naturally occurring “things,”

regardless of what they are. Thus, the Chakrabarty opinion could be read as precedent for including any genetically engineered organism

(except humans) within the scope of section 101.

Whether a court in a subsequent case will interpret Chakrabarty broadly or narrowly cannot be

predicted.
242 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Even if section 101 were interpreted as covering other genetically engineered organisms,

they probably could not be patented for failure

to meet another requirement of the patent

laws—the enablement requirement. It is generally impossible to describe a living organism in

writing with enough detail so that it can be

made on the basis of that description. Relaxing

this requirement for plants was one reason

behind the Plant Patent Act of 1930. For microorganisms, the problem is solved by depositing a

publicly available culture with a recognized national repository and referring to the accession

number in the patent. * While such an approach

may be theoretically possible for animals and insects, it may be logistically impractical. However, if tissue culture techniques advance to the

point where genetically engineered organisms

can be made from single cells and stored indefinitely in that form, there appears to be no reason to treat them any differently than microorganisms, in the absence of a specific statute

prohibiting their patentability.

● This procedure was accepted by the Court of Customs and Patent Appeals (CCPA) in upholding a patent on a process using microorganisms. Application of Argoudelis,” 434 F.Zd 1390 (CCPA 1970).

This procedure should also be acceptable for patents on microorganisms themselves.

Potential impacts of the decision and

related policy issues

During the 8-year history of the Chakrabarty

case and the surrounding public debate, numerous assertions were made about the potential impacts of permitting patents on genetically

engineered organisms. They ranged from more

immediate effects on the biotechnology industry, the patent system, and academic research

to the long-term impacts on genetic diversity

and the food supply. In addition, two major policy issues that have been raised are the morality

of patenting living organisms; and the propriety

of permitting private ownership of inventions

from publicly funded research.

Impacts on industry

The basic question for industry is the extent

to which permitting patents on genetically engineered organisms will stimulate both their development and the growth of the industries employing them. To ascertain this requires first an

examination of the theory and social policies

underlying the patent system.

THE RELATIONSHIP BETWEEN PATENTS

AND INNOVATION

The patent system is supposed to stimulate innovation—the process by which an invention is

brought into commercial use—because the inventor does not receive financial rewards until

the invention is used commercially. The Constitution itself presumes this, as do the statutes

enacted pursuant to the patent clause in article

I, section 8. Attempts have been made to subject

this presumption to empirical analysis; but innovation is extraordinarily complex and involves interacting factors that are difficult to

separate. In addition, the existence of patents

and trade secrets as alternative means for protection makes it almost impossible to study the

effects of patents alone on invention and innovation. *

● A major reuson for the lack of empirical studies has been the

lack of appropriate data. The information available on the number

of patents applied for and issued does not indicate the importance,

economic benefits, or economic costs of inventions (whether patented or unpatented) that may not have existed at all or may have

been created more slowly if not for the patent system.S In Presi-
Ch. 12—Patenting Living Organisms . 243

Several reasonable arguments have been presented to support the presumption that the patent system stimulates innovation. First, the potential for the exclusive commercialization of a

new product or process creates the incentive to

undertake the long, risky, and expensive process from research through development to marketing. At every stage of innovation—from defining priorities and making initial estimates of

an invention’s value to advertising the finished

product—the inventor and his backers must

spend time, money, and effort, not only to develop a product but to convince others of its

worth. Only a small percentage of new ideas or

inventions survive. If a competitor, particularly

a larger firm with a well-developed marketing

capability, were free to copy a product at this

point, smaller firms would have little incentive

to undertake the process of innovation.

Second, the information and new knowledge

disclosed by the patent allows others to develop

competing, and presumably better, products by

improving on the patented product or “inventing around” it. Third, patents may reduce

unnecessary costs to individual firms, thereby

freeing resources for further innovation, Once

a Patent is issued, competitors can redirect

research and development (R&D) funds into

other areas. For the firm holding the patent,

maintaining control over the technology is

theoretically less expensive, since the costs of

trade secret protection are no longer

required. * *

Anecdotal accounts support the proposition

that patents stimulate innovation; probably the

best known is the story of penicillin. Although

dent Carter’s recent report on industrial innovation, the patent

policy committee, composed of industry representatives having

long experience with the patent system, recommended ways of

enhancing innovation by improving the patent system, including

the patenting of industrially important living organisms. However,

they pro~’ided no hard economic data to support their recommendation.’

‘Carole Kitti, and Charles L. Trozzo, T’he Effects of Patent and

Antifrusl Laws, fiegulations, and Practices on innovation, vol. II (Arlington, t’a., Institute for Defense Aniilyses, 1976), pp. 2,9.

‘U.S. Department of Commerce, Advisory Committee on lndusIria/ Innovation: Final Report, September 1Y79, pp. 148-149.

● *Patent rights can be very expensive to enforce against an in-

!’ringer, however, should litigation be necessary.

Sir Alexander Fleming had discovered a promising weapon against bacterial infection, it took

him, over 10 years to get the money and facilities

he needed to purify and produce penicillin in

bulk. Only World War II and an international effort finally accomplished that task. Sir Howard

Florey, who shared the Nobel prize with Fleming for developing penicillin, attributed the

delay to their not having patented the drug,

which he termed “a cardinal error. “10

Some have claimed that the monopoly power

of a patent can be used to retard innovation. A

corporation can legally refuse to license a patent on a basic invention to holders of patents on

improvements, thus protecting its product from

becoming less attractive or obsolete. On the

other hand, unless the corporation can satisfy

the market for its product, it is usually in its

economic interest to engage in cross-licensing

arrangements with holders of improvement patents; it receives royalties and all parties can

market the improved product. Cross-licensing

has been misused several times by a few dominant firms in an attempt to exclude innovative

new firms from their markets. Such arrangements violate the antitrust laws. Whether or not

that body of law adequately prevents patent

misuse is beyond the scope of this report.

THE ADVANTAGES OF PATENTING

LIVING ORGANISMS

Given the presumed connection between

patents and innovation, the next question is

whether patenting a living organism would add

significant protection for the patent holder, or

whether alternative approaches would be sufficient. In this context, it is necessary to focus on

the present industrial applications—which involve only micro-organisms—to examine alternative forms of patent coverage and to compare

the protection offered by trade secrecy with

that offered by patents.

Opinions vary widely among spokesmen for

the genetic engineering companies on the value

of patenting micro-organisms.11 Spokesmen for

Genentech, Inc., have stated numerous times

‘“Ibid, pp. 170-171.

I ID. Dickson, “patenting Living organisms: HOW to Beat the BugRustlers, ” Nature, vol. 283, Jan. 10, 1980, pp. 128-129.
244 ● Impacts of Applied Genetics —Micro-Organisms, Plants, and Animals

that such patents are crucial to the development

of the industry, while others have stated their

preference for trade secrecy.

Genentech’s friend-of-the-court brief filed in

the Chakrabarty case stated, “The patent incentive did, and doubtless elsewhere it will, prove

to be an important if not indispensable factor in

attracting private support for life-giving research. ”l2 Genentech has also supported increased patent protection because, to attract

top scientists to the company, it had to give

assurances that they would be able to publish

freely .13 This severely curtails any reliance on

trade secrets.

The rationale behind the contrary position is

based on the belief that the industry is moving

so quickly that today’s frontrunner is not necessarily tomorrow’s, and that unique knowledge

translates into competitive advantage. Thus, in a

strategy similar to that of the advanced microelectronics industry, firms may prefer to rely

on trade secrets even for patentable inventions,

coupled with an intense marketing effort once

an invention has reached the commercial stage.

The idea is to get the jump on competitors and

to stay in front. 14

The uncertainty about whether micro-organisms could be patented before the Supreme

Court’s decision does not appear to have hindered the development of the industry. Clearly,

companies did not have any difficulty raising

capital—e.g., before the decision, Cetus Corp.

had a paper value of $250 million without holding a single patent on a genetically engineered

organism. Moreover, products such as insulin,

human growth hormone, and interferon were

being made, albeit in small quantities, by unpatented, genetically modified organisms. (See

ch. 4.)

Before the decision, companies relied either

entirely on trade secrecy for protection, or on a

combination of patents on the microbiological

process and the product and trade secret protection of the mice-organism itself. Considering

iZBrief f~r (;ellelltech as Amicus (hriae, p. ~.

‘Whonlas Kiley, t’ice President and (;eneral [kmnsel for

Genentech, personal communication, Apr. 15, 1980.

j4Di~ks~n, op cit., p. 128.

the existence of such protection, the question is

what the actual advantages are to patenting the

micro-organisms as well.

One advantage results from the ability of a

living organism to reproduce itself. Developing

a new microbe for a specific purpose, such as

the production of human insulin, can be a long,

difficult, and costly procedure. Yet once it is

developed, it reproduces endlessly, and anybody acquiring a culture would have the benefit

of the development process at little or no cost

unless the organism were patented.

Often, a company is able to keep the microbe

a trade secret, since only the product is sold.

However, where the microbe is the product—

such as with Chakrabarty’s oil-eating bacterium—patenting the organism is the best means

of protection. Moreover, even when a microbe

itself can be kept under lock and key, a company desiring to patent the process in which it is

used must place a sample culture in a public

repository to meet the enablement requirement.

A competitor could legally obtain the microorganism. If the competitor were to use it to

make the product for commercial purposes, the

company might suspect infringement but have

difficulty proving it, especially when the product is not patented, The infringing activity

would take place entirely within the confines of

the competitor’s plant. Mere suspicion is not sufficient legal grounds for inspecting the competitor’s plant for evidence of infringement

when the unpatented product could theoretically be made by many different methods

besides the one patented. *

A second, but less certain, advantage provided by patenting the micro-organism is that

even uses and products of the organism not discovered by the inventor would be protected indirectly. That is, while new uses and products

could be patented by their inventors, those patents would be “dominated” by the micro-organism patent. Royalties would have to be paid

“Some would answer this assertion by saying that ii lawsuit

coutd he stiirt~d even on I he hasis of little et’idence; the suing comPiit)~ t~ould ]WIY oil the discovery }) IWC[?SS, which is Iil]eral and

}$’iCi(?-l’illlgillg, to provide iill~ existing evid(?nce of infringenlent.
Ch. 12—Patenting Living Organisms ● 245

whenever the micro-organism was used for

commercial purposes. Whether this would be a

significant advantage in practice is uncertain.

Usually, only one product is optimally produced

by a given micro-organism and only one microorganism is best for a given process. Presumably, the micro-organism’s inventor would

also have discovered and patented its best use

and product.

Another alternative to patenting a man-made

micro-organism, besides trade secrecy, is to patent its manmade components. Examples of

these include a plasmid containing the cloned

gene, a sequence of DNA, or a synthetic gene

made by the reverse transcriptase process.

These components, which are nothing more

than strings of inanimate chemicals, would not

be unpatentable products of nature if they were

made in the laboratory and were not identical to

the natural material. Patenting them would not

be equivalent to patenting the entire organism,

since their function would be affected in varying degrees by the internal environment of their

host. Nevertheless, the inventor of a particularly useful component, such as an efficient

and stable plasmid, might want to patent it regardless of whether or not the organism could

be patented, since it could be used in an indefinite number of different micro-organisms.

Thus, if Congress were to prohibit patenting

of micro-organisms because they are alive, industry could compensate to a large degree by

patenting inanimate components. On the other

hand, if Congress allows the Supreme Court’s

decision to stand, certain components will undoubtedly still be patented. In fact, such patents

nay become more important than patents for

micro-organisms, since the components are the

critical elements of genetic engineering.

PATENT V. TRADE SECRET PROTECTION

Even with the advantages provided by patenting a micro-organism, a company could still

decide to rely on trade secrecy. In choosing between these two options, it would evaluate the

following factors: 15

“K. Saliw’atl(’tlik, “hli(’l>ol)iologi( ’ill lmwltiot)s: Prolwt 1).v l)iit(’llt-

~ 01” hlililllilill ilS il ‘1’l’il(lt’ S(>(’l’t’ll’” f}(:l’t’/f)/Jnft:r?l.s in Indms(rial

icrobiolo<qv, Id. 19, 1978, lq), 273, 277.

●

●

●

●

●

●

●

whether the organism itself or the substance that it makes will be the commercial

product,

whether there is any significant doubt of

its meeting the legal requirements for

patenting,

whether there is the likelihood of others

discovering it independently,

whether it is a pioneer invention,

what its projected commercial life is and

how readily others could improve on it if it

were disclosed in a patent,

whether there are any plans for scientific

publication, and

what the costs of patenting are versus reliance on trade secrecy.

The first two factors make the decision easy.

Obviously, an organism like Chakrabarty’s can

best be protected by a patent. In most instances,

the substance made by the organism is the commercial product, In that case, if there are significant doubts that the organism can meet all

the legal requirements for patentability, the

company would probably decide to rely on

trade secrecy.

The next three factors require difficult decisions to be made on the basis of the characteristics of the new organism, its product, and

the competitive environment. If research to develop a particular product is widespread and intense (as is the case with interferon), the risk of

a competitor developing the invention independently provides a significant incentive for

patenting. On the other hand, reverse engineering (examination of a product by experts to discover the process by which it was made) by

competitors is virtually impossible for products

of micro-organisms because of the variability

and biochemical complexity of microbiological

processes.

Thus, greater protection may often lie in

keeping a process secret, even if the microbe

and the process could be patented. This is especially true for a process that is only a minor

improvement in the state of the art or that produces an unpatentable product already made by

many competitors. The commercial life of the

process might be limited if it were patented because infringement would be difficult to detect

76-565 0 - 81 - 17
246 . Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

and not worth the time and money to prosecute

Reliance on trade secrecy might then extend its

commercial life.

Most companies would patent truly pioneer

inventions, which often provide the opportunity

for developing large markets. Moreover, patents of this sort tend to have long commercial

lives, since it is difficult to circumvent a pioneer

invention and since any improvements are still

subject to the pioneer patent. Furthermore, infringement is easy to detect because of the invention’s trailblazing nature.

The last two factors involve considerations

secondary to a product and its market. Obviously, any publication of the experiments

leading to an invention foreclose the option of

trade secrecy. Also, company must evaluate the

options of protection via either patenting or

trade secrecy in terms of their respective cost

effectiveness.

IMPACT OF THE COURT’S DECISION

ON THE BIOTECHNOLOGY INDUSTRY

The Chakrabarty decision will add some protection for microbiological inventions by providing companies with an additional incentive

for* the commercial development of their inventions, particularly in marginal cases, by lowering uncertainty and risk. A greater effect will

result from the new information disclosed in

patents on inventions that otherwise might have

been kept secret indefinitely. Competitors and

academicians will gain new knowledge as well

as a new organism upon which to build. The

Patent office had deferred action on about 150

applications, while awaiting the Court’s decision; as of December 1980, it was processing approximately 200 applications on micro-organisms. 16*

Depending on the eventual number and importance of patented inventions that would

have otherwise been kept as trade secrets, the

ultimate effect of the decision on innovation in

the biotechnology industry could be significant.

16R[+11[) ‘l’[~~llll~v~l,, ,\SSISlillll (:t)llllllissi[)ll(![’ 1’01” PillPlltS, [ 1.S. Piil -

(1111 illl(l ‘l’1.iidolliii~.k otl’i(x?, }](~t’sotliil (’otllllltllli($iit iou, Jilt]. 8, I !]& 1.

“1’11[’S(’ ill)pli(’iltiol)S i 1l(’lll(lt> ill)()(lt I ()() 011 ~[gllf]t i(’illl}r (’11-

gineerd mi(wtws illl(l iil)ollt 100” 011 (“llltlll’[~S ot” stl-;iil]s is(]lilt(~(l

1’1’0111 Ililt(ll’(’.

Conversely, if the Court had reached the opposite decision, the industry would have been

held back only moderately because of reasonably effective alternative means of protection.

Impacts of the Court's decision on the

patent law and the Patent

and Trademark Office

The key rationale supporting the Court’s

holding Chakrabarty’s microbe to be patentable

was the fact that it was manmade; its status as a

living organism was irrelevant. The Patent Office interprets this decision as also permitting

patents on micro-organisms found in nature but

whose useful properties depend on human intervention other than genetic engineering,l7

e.g., if the isolation of a pure culture of a

microbial strain induces it to produce an antibiotic, that pure culture would be patentable

subject matter.

Because of the complexity, reproducibility,

and mutability of living organisms, the decision

may cause some problems for a body of law designed more for inanimate objects than for living organisms. It raises questions about the

proper interpretation and application of the requirements for novelty, nonobviousness, and

enablement. In addition, it raises questions

about how broad the scope of patent coverage

on important micro-organisms should be and

about the continuing need for the two plant protection Acts. These uncertainties could result in

increased litigation, making it more difficult and

costly for owners of patents on living organism!

to enforce their rights.

The complexity of living matter will make i

difficult for anyone examining the invention to

determine if it meets the requirements for nov

elty, nonobviousness, and enablement. Microorganisms can have different characteristics in

different environments.’ Moreover, microbic

taxonomists often differ on the precise classification of microbial strains. Even after expensive

tests, uncertainty may still exist about whether

a specific micro-organism is distinct from othter

known strains; scientists do not have complete

‘‘1 hid, Ji~ll. 7, 19S 1.
Ch. 12—Patenting Living Organisms ● 247

knowledge of any single organism’s biophysical

and biochemical mechanisms. Consequently,

there may be cases where it is difficult to know

the prior art precisely enough to make a determination of novelty.

Similarly, microbial complexity raises problems in determining nonobviousness because

there are so many different ways of engineering

a new organism with a desired trait—e.g., a

gene could be inserted into a given plasmid at

several different positions. If a microbe with the

gene at one position in the plasmid were

patented, could a patent be denied to an otherwise structurally identical organism with the

gene at a different position because the second

was obvious? Perhaps not. The second organism

would probably not be an obvious invention if it

provided significantly more of the product, a

better quality product under similar fermenting

conditions, or the same product under cheaper

operating conditions.

As to enablement, the major problem has

been discussed previously; placing a culture of

the micro-organism into a repository is the accepted solution. One problem with repositories,

however, is their potential misuse. In a case involving alleged price fixing and unfair competition—e.g., the Federal Trade Commission found

that micro-organisms placed in a public repository pursuant to process and product patents

on the antibiotic Aureomycin did not produce

the antibiotic in commercially significant

amounts; in actual practice, other strains were

being used for production, and the company involved was able to benefit from a patent, while,

in effect, retaining the crucial micro-organism

as a trade secret. 18*

Complexity also raises questions about the appropriate scope of patent coverage. In a patent,

the inventor is permitted to claim his invention

as broadly as possible, so long as the claims

l#Anlerican (:vanamid (:0., ef. al., 6~J k“I’(; 1747, 1905 n. 14 ( 1963),

vara[cd and remanded, 363 *’. zcl 7.57 (6th (:ir. 1966), readopted 72

E-[’(: 623 ( 1967), affirmed 401 F.2d 574 f6th (;ir. 1968), cerf. denied,

394 [ 1.s. 920 ( 1 969).

“’1’he rompany had maintained that sec. I I z simply required it

to deposit ii strain that r(mfnrnwci to the des(x’iption of the one

found in the paterlt application. ti(n~,eier, it is often the ciise with

I)ii[’teriii thiit IWiIly striiins ot” it spw’ies will (wniorm to eten th~

most prcrisel)’ written description.

made do not overlap with any “prior art” or obvious extensions thereof—e.g., a person who

developed a particular strain of Escherichia coli

that produced human insulin through a genetically modified plasmid could be entitled to a patent covering all strains of E. coli that produce

the insulin in the same way. Chakrabarty’s patent application—e.g., claimed “a bacterium from

the genus pseudomonas containing therein at

least two stable energy generating plasmids,

each of said plasmids providing a separate

hydrocarbon degradative pathway.” Several

species and hundreds of strains of Pseudomonas

fit this description. A patent limited to a particular microbial strain is not particularly

valuable because it can easily be circumvented

by applying the inventive concept to a sister

strain; on the other hand, a patent covering a

whole genus of micro-organism (or several) may

retard competition. This problem will probably

be resolved by the Patent Office and the courts

on a case-by-case basis.

Another aspect of the same problem is

whether a patent on an organism would cover

mutants. It would not if the mutation occurred

spontaneously and sufficiently altered the

claimed properties. However, if a new organism

were made in a laboratory with a patented

organism as a starting point, the situation would

be analogous to one where an inventor can patent an improved version of a machine but must

come to terms with the holder of the “dominant” patent before marketing it.

The Chakrabarty decision also raises questions about the scope of section 101 and its relation to the plant protection Acts—e.g., plant

tissue culture is, in effect, a collection of microorganisms; should it be viewed as coming under

section 101 instead of either of the plant protection Acts? Could plants excluded under these

Acts—such as tuber-propagated plants or firstgeneration hybrids—be patented under section

101? Could any plants or seeds be patented

under section 101, and if so, is there still a need

for the plant protection Acts? If there is a need,

would the Acts be administered better by only

one agency? The Senate Committee on Appropriations has directed the Departments of Commerce and Agriculture to submit a report
248 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

within 120 days of the Chakrabarty decision on

the advisability of shifting the examining function to USDA. 19 As of December 1980, this issue

was still under study. These questions could be

resolved by the courts, but they are probably

more amenable to a statutory solution.

Another effect of the decision could be on

patent enforcement. The various uncertainties

discussed above may have to be resolved

through costly litigation. Moreover, in specific

cases, the problems associated with describing a

micro-organism in sufficient detail may increase

the chances that a patent will be declared

invalid. In any event, litigation costs would

probably increase as more expert testimony is

needed.

The fact that organisms mutate might introduce still another complication into infringement actions. A deposited micro-organism is the

standard by which possible infringement would

be judged. If it has mutated with respect to one

of its significant characteristics, a patent holder

who is seeking to prove infringement may have

no case. While this problem does not appear to

be amenable to a statutory solution, the risk of

such a mutation is actually quite small. *

Because a living invention reproduces itself,

the statutory definition of infringement may

have to be changed. Presently, infringement

consists of making, using, or selling a patented

invention without the permission of the patent

holder. Theoretically, someone could take part

of a publicly available micro-organism culture,

reproduce it, and give it away. Arguably, this is

not “making” the invention, and the patent

holder would have the burdensome and expensive task of going after each user. The two plant

protection statutes deal with this problem by

specifically prohibiting unauthorized reproduction of the protected plant. This approach

may be necessary for other living inventions,

How all of these uncertainties will affect the

Patent Office’s processing of applications cannot

be predicted. Currently, the average processing

time for all applications is 22 months; separate

j~s. R~P~. No. 96.251, 96th (king. 1s1 Sess., 1979, P. 46.

“Most nlicro-or#misms can be stored in ii freeze-dried form,

which ~ntilils virtually II(J risk (It’ n]tltiiti[)n.

information on genetic engineering applications

is not available.20 It may take examiners longer

to process applications on micro-organisms than

for those covering only microbiological processes or products because of the interpretive

problems mentioned. Moreover, the Patent Office will have to develop greater expertise in

molecular genetics—a frontier scientific field

that has only recently been the subject of patent

applications. On the other hand, the Office

generally faces this problem for any new area

of technology.

In terms of increased numbers of applications, the decision is not expected to have a significant effect on the Patent Office operations in

the next few years. The Office receives approximately 100,000 applications a year, and it has

about 900 examiners, each processing an average of about 100 applications per year. Figures

on the number of applications on genetically

engineered organisms vary, depending on how

the category is defined, and precise information

has not been tabulated by the Patent Office.

Rough estimates indicate that in February 1980

about 50 applications were pending, and by

December 1980, that number had increased to

about 100. Applications are being filed at the

rate of about 5 per month. Also, just over 100

are pending on microbes that have been isolated

and purified from natural sources, but have not

been genetically engineered. Four examiners

are working on both categories as well as

others. Thus, in view of the total operations of

the office, these applications require only a

small part of its resources. over the next few

years, the number is expected to increase because of the decision and developments in the

field but not to a point where more than a few

additional examiners will be needed.2l

Impact of the Court's decision on

academic research

Many academicians have voiced concerns

about the effects on research of the Chakrabarty

decision and the commercialization of molecular biology in general. They claim that the reZot+elle Tegtfllever, person~l c(~lllnllttli~:ati[)ll, Dti:. 1.’, 198(J.

Zllt)id, , De[; . 151 1980, iind Jiil). 8, 1981.
Ch. 12—Patenting Living Organisms ● 249

suits of rDNA research are not being published

while patent applications are pending, discussion at scientific meetings is being curtailed, and

novel organisms are less likely to be freely exchanged. A related concern is that scientific

papers may not be citing the work of other scientists to avoid casting doubt on the novelty or

inventiveness of the author’s work, should he

decide to apply for a patent. Finally, there is

concern that the granting of patents on basic

scientific processes used in the research laboratory will directly impede basic research—e.g.,

two scientists have recently been granted a patent on the most fundamental process of molecular genetic technology—the transfer of a gene

in a plasmid using rDNA techniques.22 The patent has been transferred to the universities

where they did their work—Stanford and the

University of California at San Francisco (UCSF).

Although both universities have stated they

would grant low-royalty licenses to anyone who

complied with the National Institutes of Health

(NIH) Guidelines, subsequent owners of fundamental process patents may not be so

altruistic.

There are several reasons for believing that

these concerns, although genuinely held, are

somewhat overstated. First, patents on fundamental scientific processes or organisms should

not directly hinder research. The courts have

interpreted patent coverage as not applying to

research; in other words, the patent covers only

the commercial use of the invention.23 Also, it

would be difficult and prohibitively expensive

for a patent holder to bring infringement actions against a large number of geographically

separated scientists. Second, patents ultimately

result in full disclosure. If patents were not

available, trade secrecy could be relied on, with

the result that important information might

never become publicly available. Third, although delays occur while a patent application

is pending, they often happen anyway while experiments are being conducted or while articles

ZZ[ 1s, f~iitt?l]t No. 4,23i’,22LI, issued [](’[’. 2, 198[).

z~Kaz ,Man@act[lrjng (1]. v, Chcsehrough-Ponds, Inc., 211 ~’. SUPP.

815 (S.l). N.Y. 1962) [dictum), +j_irmed 317 F.2d 679 (2cI [:ir. 1963);

(,’hesterj_ie/d t’. (hi[ed States, 159 h-. Supp. 371 [(It. (:1. 1958); fhgan

t’. I,ear A via, 55 F. Supp. 223 (S. [). N.Y. 1944) (dictum); Akro Aga(e

(,’o. t. Afas[er Marb/e (,’o., 18 F. Supp. 305 (N.l). JV. \’a. lY37).

are being prepared for publication because of

the competitive nature of modern science.

Essentially, the issue is the effect of the commercialization of research results on the research process itself. Even if patents were not

available for biological inventions, the inventor

would simply keep his results secret if he were

interested in commercialization. Viewed from

this perspective, it is difficult to see why the

availability of patents should affect the exchange of scientific information in genetic research any more than it does in any other field

of research with commercial potential. The

Chakrabarty decision may inhibit the dissemination of information only if it creates an atmosphere that stimulates academic scientists to

commercialize their findings. However, if it encourages them to rely on patents rather than on

trade secrets, it will ultimately enhance the

dissemination of information.

Impacts of the Court’s decision on

genetic diversity and the food supply

Some public interest groups have claimed

that patenting genetically modified organisms

will adversely affect genetic diversity and the

food supply. The claim is based on an analogy to

a situation alleged to exist for plants. Briefly, the

groups claim that patenting micro-organisms

will irrevocably lead to patents on animals,

which will have the same deleterious effects on

the animal gene pool and the livestock industry

as the two plant protection Acts have had on the

plant gene pool and the plant breeding industry.

The alleged effects are: loss of germplasm resources as a result of the elimination of thousands of varieties of plants; the increased risk of

widespread crop damage from pests and diseases because of the genetic uniformity resulting from using a single variety; and the increasing concentration of control of the world’s food

supply in a few multinational corporations

through their control of plant breeding companics.24

only limited evidence is available, but no conclusive connection has been demonstrated bezd~l.itlf for the peoples’ Blujincss (;(mmission as Am icus (M-he,

pp. 6-13, Iliamondv. Chakrabarf-v, 100 S. [1. 2204 (1980).
250 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

tween the plant protection laws and the loss of

genetic diversity, the encouragement of using a

single variety, and any increased control by a

few corporations of the food supply. (For a detailed discussion, see ch. 8.) Therefore, any connection between patenting micro-organisms

and potential detrimental impacts on the livestock industry appears tenuous at best. The

assumptions that the Chakrabarty decision will

inevitably lead to patenting animals, and that

the consequences will be the same as those

claimed to result from granting limited ownership rights to varieties of plants, are speculative.

The morality of patenting living

organisms

The moral issue is difficult to analyze because

it embodies at least three overlapping questions:

whether it is moral to grant exclusive rights of

ownership to a living species; whether patents

on lower forms of life will inevitably lead to

genetic engineering of humans; and whether

patenting organisms undermines the generally

held belief in the uniqueness and sanctity of life,

especially human life.

It is difficult to assess the extent of the belief

that patenting living organisms is intrinsically

immoral, and no such assessment has been

done. Its extent and intensity will probably be

directly correlated with the complexity of the

organism involved. Fewer people will be disturbed about patenting micro-organisms than

about patenting cattle. A belief in the immorality of patenting a living organism is a value judgment to which Congress may wish to give some

consideration.

The second aspect of the moral issue revolves

around the well-known metaphor of the “slippery slope”-the fear that the first steps along

the path of genetic engineering may irrevocably

lead to man. Technology, at times, appears to

have its own momentum; the aphorism “what

can be done, will be done” has been true in the

past. Thus, some people fear that patenting

micro-organisms may indeed set a dangerous

precedent and encourage the technology to progress to the point of the ultimate dehumanization—the engineering of people as an industrial

enterprise. 25

The Chakrabarty opinion was written in narrow terms. But while its reasoning might be applied to a future case involving an animal or insect, it simply could not be used to justify the

patenting of human beings because of the 13th

amendment to the Constitution, which prohibits

the ownership of humans.

One way to negotiate the slippery slope is to

deal directly with the adverse aspects of the

technology. Barriers can be erected along the

slope; the Constitution already protects

humans. Congress can erect other barriers by

statute, specifically drawing lines as to which

organisms can or cannot be patented.

The third part of the issue is religous or

philosophical in nature. For many, the patenting of a living organism undermines the awe

and deep respect they hold for the unique nature of life. Moreover, it raises apprehensions of

an ultimate threat to concepts of the nature of

humanity and its place in the universe. To these

people, if life can be engineered and patented,

perhaps it is not special or sacred. If this is true

of lower organisms, why would human beings

be different? (This and other aspects of the

morality issue are discussed in greater detail in

ch. 13.)

Private ownership of inventions

from publicly funded research

Much of the basic research in molecular genetics has been funded by Federal grants. Most

of the work leading to the development of rDNA

techniques—e.g., was performed at Stanford

University and UCSF under NIH grants. The

scientists involved have received a patent on

that fundamental scientific process. Some opponents of patenting organisms have argued

that private parties should not be permitted to

own inventions resulting from federally funded

R&D; and in any event, there is something

special about molecular genetics that requires

the Federal Government to retain ownership of

Zslhid., p. 25.
Ch. 12—Patenting Living Organisms ● 251

federally funded inventions and to make them

generally available through nonexclusive

licenses.

Until recently, there had been no comprehensive, governmentwide policy regarding ownership of patents on federally funded inventions.

Some agencies, such as the Department of

Health and Human Services (DHHS), permitted

nonprofit institutional grantees to own patents

on inventions (subject to conditions deemed

necessary to protect the public interest) if they

had formal procedures for administering them.

However, most agencies generally retained title

to such patents, making them available to anyone in the private sector for development and

possible commercialization through nonexclusive licenses.

The rationale behind the policy was simply

that inventions developed by public money

should be available to all—including private industry—on a nonexclusive basis. This arrangement had been criticized as not providing sufficient incentive for industry to take the risks to

develop the inventions. Of the more than 28,000

patents owned by the Government, less than 4

percent have been successfully licensed; on the

other hand, universities, which do grant exclusive licenses on patents that they own, have

been able to license 33 percent of their

patents. 26

On December 12, 1980, President Carter

signed the Government Patent Policy Act of

1980. The Act sets forth congressional policy

that the patent system be used to promote the

utilization of inventions developed under federally supported R&D projects by nonprofit

organizations and small businesses. To this end,

the organization or firm may elect to retain title

to those inventions, subject to various conditions designed to protect the public interest.

Such conditions include retention by the funding agency of a nonexclusive, irrevocable, paidup license to use the invention, and the right of

the Government to act where efforts are not

being made to commercialize the invention, in

cases of health or safety needs, or when the

use of the invention is required by Federal regulations.

z~s, ~q)t N(). ~~.~~(), 961 h (:ong. 1$1 S~SS, I :)~!], l). z

There is still the question of whether patents

on molecular techniques or genetically engineered micro-organisms are sufficiently different to merit exception from any general patent policy decided on by Congress. For some,

the molecular genetic techniques are unique because they are powerful scientific tools that can

manipulate the life processes as never before.

However, in a November 1977 report, NIH took

the following position with regard to patents on

rDNA inventions developed under DHHS-NIH

support: 27*

There are no compelling economic, social, or

moral reasons to distinguish these inventions

from others involving biological substances or

processes that have been patented, even when

partially or wholly developed with public funds.

The report was prompted by the StanfordUCSF patent application. Even though the application was in accord with the funding agreements between the institutions and NIH, the

universities requested a formal NIH opinion on

the issue in view of the intense public interest in

rDNA research. NIH solicited comments from a

group of approximately 67 individuals, ranging

from academic and industrial scientists to

students, lawyers, and philosophers. 28 T h e

review and analysis of the responses were

referred to the Federal Interagency Committee

on rDNA Research, the Public Health Service,

and the Office of the General Counsel of the Department of Health, Education, and Welfare

(now DHHS). A fairly uniform consensus on the

above-quoted finding developed in this process;

the one significant dissenter, the Department of

Justice, contended that the Government should

retain ownership of any invention resulting

from federally funded rDNA research because

of the great public interest in that research.

zTThe patenting Of n~omlhinant IJVA Research Inventions [leveloped under DHEW Support: An Anal.vsis b-v the IIirertor, )Vationai

Institutes of Heahh, November 1977, p. 16.

● “I”he repori further concluded that no change was Iwressary in

the hasic NIH policy permitting nonprotil ()]’~iini~ii[i()lls to own

~illelltS 011 intentions developed Uilder (’olltl’ii(”tS (If’ ~1’illllS !1’0111

the Department of Health, Edll(:iltion, illld tV’[’lt’ar(’ (n(n~ 1)}{}{ S),

subject to several conditions to protect the puhl ic interest. ‘1’he

0111.V rcc’(]llln][~ilcl[!{l (’hii[l~[~ \\Ias I hitt the f(]t.]l]iil i)g]’~?~nl~l)(s IYtwo~]] Nl}l iind the institutions h(> iin~etld[?[l 10 r(quire thitt ii[lv

licens(~es ot” i[lstitutioniil l)iit[~tlt holders COIIIply with the [.~ntiii]lI]l[?nt stiin(iii[-ds 01” th(’ NIH (;ui(l(’lines in iinav production” 01’ LISP of

I’I)N/l IIIOIW’LIIPS L] II(iPI> t h(’ I h’ense ii~r[~(~n}otlt.

zn[t)i(j ,, a])~). I , I)J) 5-8’
252 . Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

Issue and Options

ISSUE: To what extent could Congress

provide for or prohibit the patentability of living organisms?

In its Chakrabarty opinion, the Supreme

Court stated that it was undertaking only the

narrow task of determining whether or not

Congress, in enacting the patent statutes, had

intended a manmade micro-organism to be excluded from patentability solely because it was

alive. Moreover, the opinion specifically invited

Congress to overrule the decision if it disagreed

with the Court’s interpretation.

Congress has several options. It can act to resolve the questions left unanswered by the

Court, overrule the decision, or develop a comprehensive statutory approach. Most importantly, Congress can draw lines; it can decide which

organisms, if any, should be patentable.

OPTIONS

A: Congress could maintain the status quo.

Congress could choose not to address the

issue of patentability and allow the law to be

developed by the courts. The advantage of this

option is that issues will be addressed as they

arise in the context of a tangible, nonhypothetical case. Some of the issues raised in the

debate on patenting may turn out to be irrelevant as the technology and the law develop.

Moreover, many of the uncertainties raised by

the Chakrabarty decision regarding provisions

of the patent law other than section 101 may be

incapable of statutory resolution. The complexity of living organisms and the increase in knowledge of molecular genetics will raise such broad

and varied questions that legal interpretations

of whether a particular biological invention

meets the requirements of novelty, nonobviousness, and enablement will best be done on a

case-by-case basis by the Patent Office and the

Federal courts.

There are two disadvantages to this option.

First, a uniform body of law may take time to

develop, since judicial decisions about new legal

questions by different Federal courts may initially conflict. Second, the Federal judiciary is

not designed to take sufficient account of the

broader political and social interests involved.

B: Congress could pass legislation dealing with

the specific legal issues raised by the Court's

decision.

Many of the legal questions do not readily

lend themselves to statutory resolution. However, three questions are fairly narrow and

well-defined and may therefore be better resolved by statute: 1) Is there a continuing need

for the plant protection Acts if plants can be

patented under section 101? 2) If there is a continuing need for these Acts, could they be administered better by one agency? 3) Should the

definition of infringement be clarified by

amending section 271 of the Federal Patent

Statutes (title 35 U. S. C.) to include reproduction

of a patented organism for the purpose of selling it?

Congressional action to clarify these issues

would provide direction for industry and the

Patent Office, and it would obviate the need for

a resolution through costly, time-consuming litigation. Lessening the chances of litigation or

the chances of a patent being declared invalid

will provide some stimulation for innovation by

lessening the risks in commercial development.

In addition, Congress could determine that the

plant protection Acts could be better administered by one agency or should be incorporated

under the more general provisions of the patent

law; if so, some administrative expenses probably could be saved.

C: Congress could mandate a study of the plant

protection Acts.

Two statutes, the Plant Patent Act of 1930

and the Plant Variety Protection Act of 1970,

grant ownership rights to plant breeders who

develop new and distinct varieties of plants.

They could serve as a model for studying the

broader, long-term potential impacts of patenting living organisms. An empirical study of the

impacts of the plant protection laws has not

been done. Such a study would be timely, not
Ch. 12—Patenting Living Organisms . 253

only because of the Chakrabarty decision, but

also because of allegations that the Acts have encouraged the planting of uniform varieties, loss

of germplasm resources, and increased concentration in the plant breeding industry. In addition, information about the Acts’ affect on innovation and competition in the breeding industry would be relevant to this aspect of the

biotechnology industry. However, it may be extremely difficult to isolate the effects of these

laws from the effects of other factors.

D: Congress could prohibit patents on any living

organism or on organisms other than those

already subject to the plant protection Acts.

By prohibiting patents on any living organisms, Congress would be accepting the

arguments of those who consider ownership

rights in living organisms to be immoral, or who

are concerned about other potentially adverse

impacts of such patents. Some of the claimed

impacts are: 1) patents would stimulate the development of molecular genetic techniques,

which will eventually lead to human genetic engineering; 2) patents contribute to an atmosphere of increasing interest in commercialization, which will discourage the open exchange

of information crucial to scientific research; and

3) plant patents and protection certificates have

encouraged the planting of uniform varieties,

loss of germplasm resources, and increasing

concentration in the plant breeding industry.

Also, by repealing the plant Acts, Congress

would be reversing the policy determination it

made in 1930 and in 1970 that ownership rights

in novel varieties of plants would stimulate

plant breeding and agricultural innovation.

A prohibitory statute would have to deal with

those organisms at the edge of life, such as

viruses. Although there are uncertainties and

disagreements in classifying some entities as

living or nonliving, Congress could be arbitrary

in its inclusions and exclusions, so long as it

clearly dealt with all of the difficult cases.

This statute by itself would slow but not stop

the < development of molecular genetic techniques and the biotechnology industry because

there are several good alternatives for maintaining exclusive control of biological inventions:

maintaining organisms as trade secrets; patenting microbiological processes and their products; and patenting the inanimate components

of a genetically engineered micro-organism,

such as plasmids, which are the crucial elements of the technique anyway. The development would be slowed primarily because information that might otherwise become public

would be kept as trade secrets. A major consequence would be that desirable products would

take longer to reach the market. Also, certain

organisms or products that might be marginally

profitable yet beneficial to society, such as some

vaccines, would be less likely to be developed.

In such cases, the recovery of development

costs would be less likely without a patent to

assure exclusive marketing rights.

Alternatively, Congress could overrule the

Chakrabarty decision by amending the patent

law to prohibit patents on organisms other than

the plants covered by the two statutes mentioned in option C. This would demonstrate

congressional intent that living organisms could

be patented only by specific statute and alleviate

concerns of those who fear the “slippery slope. ”

E: Congress could pass a comprehensive law

covering any or all organisms (except

humans).

This option recognizes the fact that Congress

can draw lines where it sees fit in this area. It

could specifically limit patenting to micro-organisms or encourage the breeding of agriculturally important animals by granting patent

rights to breeders of new and distinct breeds.

Any fears that such patents would eventually

lead to patents on human beings would be unfounded, since the 13th amendment to the constitution, which abolished slavery, prohibits

ownership of human life.

The statute would have to define included or

excluded species with precision. Although there

are taxonomic uncertainties in classifying organisms, Congress could arbitrarily include or

exclude borderline cases.

A statute that permitted patents on several

types of organisms could be modeled after the

Plant Variety Protection Act—e.g., it should

cover organisms that are novel, distinct, and
254 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

uniform in reproduction; such terms would

have to be defined, Infringement should include

the unauthorized reproduction of the organism-although reproduction for research

should be excluded to allow the development of

new varieties. In fact, consideration should be

given to covering in one statute plants and all

other organisms that Congress desires to be patentable. This would provide the advantage of

comprehensiveness and uniform treatment; it

could also address the problems discussed

under option B.

The impact of this law cannot be assessed

precisely. A comprehensive statute would stimulate the development of new organisms and

their products and would encourage dissemination of technical information; however,

such a statute is not essential to the development of the biotechnology industry, since

incentives and alternative means for protection

already exist. The secondary impacts on society

of the legislation are even harder to assess

because of the scarcity of data from which to

draw conclusions. The policy judgments will

have to be made by Congress after it weighs the

opinions of the various interest groups.

Through legislation, Congress has the chance to

balance competing views on this controversial

issue and, if necessary, to alleviate the primary

concerns about the long-term impacts of the

decision–that higher organisms will inevitably

be patented.
Chapter 13

Genetics and Society
Chapter 13

Page

Genetics and Modern Science . . . . ...........257 The “Public” and “Public Participation” . . . . . . . .261

Special Problems Posed by Genetics . .........258 Issues and Options . . . . . . , . . . . . . . . . . . . . . . . 261

science and Society. . . . . . . . . . . . . . . . . . . . .. .259 Bibliography: Suggested Further Reading , . . . . .26.5
Chapter 13

Genetics and Society

Genetics and modern science

In 1979, the organization for Economic Cooperation and Development (OECD)* published a

survey of mechanisms for settling issues involv-

.

ing science and technology in its member countries. I The OECD report noted that:2

Science and technology . . . have a number of

distinguishing characteristics which cause special problems or complications. One is ubiquity:

they are everywhere. They are at the forefront

of social change. They not only serve as agents

of change, but provide the tools for analyzing

social change. They pose, therefore, special 3.

challenges to any society seeking to shape its

own future and not just to react to change or to

the sometimes undesired effects of change.

After surveying member countries, OECD

identified six factors that distinguish issues in

science and technology from other public controversies.

1.

2.

The rapidity of change in science and technology often leads to concern. The science

of genetics is one of the most rapidly expanding areas of human knowledge in the

world todayv. And the technology of genetics

is causing quick and fundamental changes

on a variety of fronts. The news media

have consistently reported developments

in genetics, often with front-page stories.

Consequently, the public has become in- 4. creasingly aware of developments in genetics and genetic technologies and the speed

with which knowledge in the field is gathered and applied.

Many issues in today's science and technology are entirely new. Protoplasm fusion, re-

“’1’he members of’ OE(;D are: Auslra]ia, Austria, Belgium,

(;ilni]da, l)enmark, Finland, France, West Germany, Greece, lceIand, lr~liind, Italy, Japan, I,uxemhourg, the Netherlands, New

Zeiiliind, Norway, Porttl~i\l, Spain, Sweden, Switzerland, Turkey,

[he (fnited Kingdom, iitld the LJnited States.

1( ;uild K. Nichols, ‘1’echnologv on Trial: Public Participation in Decision-Makin~ Related to Science and Technology [Paris: organization for kkwnomic (kmperation and Development, 1979).

‘lt~id., p. 16.

combinant DNA (rDNA), gene synthesis,

chimeras, fertilization of mammalian embryos in vitro, and the successful introduction of foreign genes into mammals were

the subjects of science fiction until a few

years ago. Now they appear in newspapers

and popular magazines. Yet the genera]

public’s understanding of these phenomena is limited. It is difficult for people to

evaluate competing claims about the dangers and benefits of this new technology.

The scale, complexity, and interdependence

among the technologies are greater than

people suspect. As in other fields, applications of biological technology often depend

on parallel developments in areas that provide critical support systems. Breakdowns

in these systems are often as limiting as failures in the new technology itself. In other

parts of this report for example, sophisticated breeding systems in farm animals

and large-scale fermentation processes for

single-cell cultures are described. Besides

the biological technology required to support these systems, precise computerized

operations are required to ensure purity,

safety, and process control in fermentation

and to provide the population statistics

necessary for breeding decisions.

Some scientific and technological achievements may be irreversible in their effects.

Because living organisms reproduce, some

fear that it will be impossible to contain

and control a genetically altered organism

that finds its way into the environment and

produces undesirable effects. Scenarios of

escaping organisms, pandemics, and careless researchers are often drawn by critics

of today’s genetics research. The intentional release of recombinant organisms into

the environment is a related issue that will

need to be resolved in the future.

Another example of irreversibility,

brought about by the demands placed on

257
258 ● Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

5.

6.

world resources, is the accelerating loss of

plant and animal species. Concern over this

depletion of the world’s germplasm arises

because genetic traits that might meet as

yet unknown needs are being lost.

There exist strong public sensibilities about

real or imagined threats to human health.

Mistrust of experts has been stimulated by

such events as the accident at the ThreeMile Island nuclear plant and the burial of

toxic chemical wastes in the Love Canal.

Regardless of the real dangers involved,

the public’s perception of danger can be a

significant factor in decisionmaking. At

present, some perceive genetic technologies as dangerous.

A challenge to deeply held social values is being raised by scientific and technological isSpecial problems posed by

Genetics is just one among several disciplines

of the biological sciences in which major advances are being made. other areas, such as

neurobiology, behavior modification, and sociobiology, arouse similar concerns.

Genetics differs from the physical sciences

and engineering because of its intimate association with people. The increasing control over

the characteristics of organisms and the potential for altering inheritance in a directed fashion

is causing many to reevaluate themselves and

their role in the world. For some, this degree of

control is a challenge, for others, a threat, and

for still others, it causes a vague unease. Different groups have different reasons for embracing or fearing the new genetic technologies.

Religious, political, and ethical reasons have

been advanced to support different viewpoints.

The idea that research in genetics may lead

some day to the ability to direct human evolution has caused particularly strong reactions.

One reason is that such capability brings with it

responsibility for retaining the genetic integrity

of people and of the species as a whole, a responsibility formerly entrusted to forces other

than man.

sues. The increasing control over the inherited characteristics of living things causes

concern in the minds of some as to how

widely that control should be exercised

and who should be deciding about the

kinds of changes that are made. Furthermore, because genetics is basic to all living

organisms, technologies applicable to lower forms of life are theoretically applicable

to higher forms as well, including human

beings. Some wish to discourage applications in lower animals because they fear

that the use of the technologies will progress in increments, with more and more

complex organisms being altered, until human beings themselves become the object

of genetic manipulation.

genetics

Others find the idea of directing evolution exciting. They view the development of genetics

technologies in a positive light, and see opportunities to improve humanity’s condition.

They argue that the capability to change things

is, in fact, a part of evolution.

Religious arguments on both sides of this

challenge have been made. Pope John Paul 11

has decried genetic engineering as running

counter to natural law. On the other hand, one

Catholic philosopher has written:3

. . . We have always said, often without real

belief, that we were and are created by God in

His own image and likeness, “Let us make man

in our image, after our likeness” logically means

that man is by nature a creator, like his Creator.

Or at least a cocreator in a very real, awesome

manner. Not mere collaborator, nor administrator, nor caretaker. By divine command we

are creators. Why, then, should we be shocked

today to learn that we can now or soon will be

able to create the man of the future? Why

should we be horrified and denounce the sci3ROheI’t ‘1’. t+all(!O(?U1’, “We [kin-We MUSI: Reflections on the

‘1’e(~tlll[)l(~~i~ill lnlpentive)” Theological Studies 33:3, September

1972, p. 429 ;Illd ilt toot note 2.
Ch. 13—Genetics and Society . 259

entist or physician for daring to “play God?” IS it

because we have forgotten the Semitic (biblical)

conception of creation as God’s ongoing collaboration with man? Creation is our God-given

role, and our task is the ongoing creation of the

yet unfinished, still evolving nature of man.

Man has played God in the past, creating a

whole new artificial world for his comfort and

enjoyment. Obviously we have not always displayed the necessary wisdom and foresight in

that creation; so it seems to me a waste of time

and energy for scientists, ethicists, and laymen

alike to beat their breasts today, continually

pleading the question of whether or not we

have the wisdom to play God with human nature and our future. It is obvious we do not, and

never will, have all the foresight and prudence

we need for our task. But I am also convinced

that a good deal of the wisdom we lack could

have been in our hands if we had taken seriously our human vocation as transcendent creaScience and society

The public’s increasing concern about the effects of science and technology has led to demands for greater participation in decisions on

scientific and technological issues, not only in

the United States but throughout the world.

The demands imply new challenges to systems

of representative government; in every Western country, new mechanisms have been devised for increasing citizen participation. An increasingly informed population, skilled at exerting influence over policymakers, seems to be a

strong trend for the future. The media has

played an important role in this development,

reporting both on breakthroughs in science and

technology and on accidents, pollution, and the

side-effects of some technologies.

one result has been the growing politicization of science and technology. While perhaps

misunderstanding the nature of science as a

process, the public has become disenchanted by

recent accidents associated with technology, by

experts who openly disagree with one another,

and by the selective use of information by some

scientific supporters to obtain a political objective. The public has seen that technology affects

tures, creatures oriented toward the future

(here and hereafter), a future in which we are

cocreators.

Genetics thus poses social dilemmas that most

other technologies based in the physical sciences do not. Issues such as sex selection, the

abortion of a genetically defective fetus, and in

vitro fertilization raise conflicts between individual rights and social responsibility, and

they challenge the religious or moral beliefs of

many. Furthermore, people sense that genetics

will pose even more difficult dilemmas in the future. Although many cannot fully articulate the

basis for their concern, considerations such as

those discussed in this section are cited. The

strong emotions aroused by genetics and by the

questions of how much and what kind of research should be done are at least partly rooted

in deeply held human values.

the distribution of benefits in society; it can

have unequal impacts, and those who pay or

who are most in need are not necessarily always

those who benefit.

A national opinion survey of a random sample of 1,679 U.S. adults conducted for the National Commission for the Protection of Human

Subjects of Biomedical and Behavioral Research4

made clear that there is public doubt concerning equity. Sixty percent of those polled felt that

new tests and treatments deriving from medical

research are not equally accessible to all Americans. Seventy percent felt that those most likely

to benefit from a new test or treatment of limited availability were those who could pay for it

or who knew an important doctor. This should

be compared with the 85 percent who felt that a

new test or treatment should be available to

those who apply first or who are most in need.

4’4S~)[W’iill SIU(IJ’, IIllpli(’iitioll!i 01” ,ll(lt’illl(’t?$i iil f}ioll)fl(li(’ill illl[l B(’-

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( ‘(mltllissioll t“ol” Illf> l}lx)t(~(liotl ot” Illllllilll Slll)jo(’ls ot” lliolll(~(li(’ill

ilt~(l l\(*ilit\’iol. ii I K(’~f~iit.(’11, I III I“;\\’ I)(ll)li(’iil ion N’(). (OS) 78-()() 15.
260 ● Impacts of Applied Genetics—Micro-organisms, Plants, and Animals

Public concern and demand for involvement

in the policy process is illustrated by the response of communities to plans for laboratories

that would conduct rDNA research. Perhaps the

best known example is Cambridge, Mass.,

where plans were announced for construction

of a moderate containment laboratory at Harvard University. Concern over this facility led to

the formation of the Cambridge Experimentation Review Board (CERB). Composed of nine citizens—all laymen with respect to rDNA research—the CERB spent 6 months studying the

subject and listening to testimony from scientists with opposing points of view. Their final

recommendations did not differ substantially

from the NIH Guidelines; but the process was

crucial. CERB demonstrated that citizens could

acquire enough knowledge about a highly technical subject to develop realistic criteria and apply them. Similar responses to proposed laboratories have occurred in a number of other

American communities, including Ann Arbor,

Mich., and Princeton, N.J.5

These reactions, and similar phenomena surrounding controversies like nuclear power, indicate that the desire for citizen participation is

strong and widespread. Recognizing this, each

Federal agency has its own rules and mechanisms for citizen input. Special ad hoc commissions are sometimes formed to collect information from private citizens before decisions

are made on particular projects. Congressional

hearings held around the country and in Washington, D. C., are perhaps the best known of

these inquiries. While these mechanisms sometimes slow the decisionmaking process, they

help legitimize some decisions, and their role

will probably expand in the future.

In corporate science and technology, public

demands are being felt as well. Present regulations for environmental protection and worker

and product safety have significantly altered

‘Richi~rd Hulton, Bio-l?evolution.’ DNA and Ihe Ethics of ManMade L;fe [New York: New Atllel.it’iill I,ihrary (Me[~tolT), 1978).

corporate research and development efforts.

The public is also becoming more involved in

corporate decisionmaking—e. g., through “’public accountability” campaigns by stockholders to

influence company policies.

With the politicization of science, the process

of research itself is coming under increasing

public scrutiny–most recently in cases of possible biohazards, research with human subjects,

and research on fetuses. Some efforts are underway to require better treatment of research

animals as well.

The relationship between science and society,

between human beings and their tools, is a constantly evolving one. The process that has been

called the “dialogue within science and the dialogue between the scientific community and the

general public"6 will continue to search for

standards of responsibility. It is likely that as

long as science remains as dependent on public

funds as it has over the past 40 years, it will be

held accountable to public values. As has been

noted: 7

The technologies of war, industrialization,

medicine, environmental protection, etc., appear less as the demonstrations of superior

claims of knowledge and more and more as the

symbols of the ethical and political choices underlying the distribution of the power of scientific knowledge among competing social values . . . . This cultural shift of emphasis from the

role of science in the intellectual construction of

reality to the role of science in the ethical construction of society may indicate a profound

transformation in the parameters of the social

assessment of science and its relations to the political order.

6Diitli(d (~iillilhiill, ‘{t:thicid Re$pwlsibili[y in Science in Ihe F’itce

of Llllcel’lilill (~ollse[[llell{;es,’” Ethical and Scientfic Issues Posed by

Human [Jses qf Mo/ecular Genetics, Mitts~ [.appe and Robert S.

Morison” k!ds. ), Atltliils of” the New Y~rsk A(;iidellly of Sciences MS,

Jiii]. 23, 1976, ~). I(k

‘Yiit’oil fikf’iihi, “’1’he Politics (Jt’ the !$~~iill Assessment of !$cience”

in ‘/’he Socia/ Assessment of Science, b;. Mendelson, D. Nelkin, P.

Weit}gilimt (eds. ), (lmlerence Proceedings (Bielefeid, West (;erll~iitl~: A&W opilz, 1978), p. 181.
Ch. 13—Genetics and Society . 261

The “public’) and ‘(public participation”

These are terms with vastly different meanings to different people. Some take “the public”

to mean an organized public interest group;

others consider such groups the “professional”

public and feel they have agendas that differ

from those of the less organized “general” public. As OECD stated:8

Public participation is a concept in search of a

definition. Because it means different things to

different people, agreement on what constitutes

“the public” and what delineates “participation”

is difficult to achieve. The public is not of course

homogeneous; it is comprised of many heterogeneous elements, interests, and preoccupations. The emergence over the last several decades of new and sometime vocal special interest

groups, each with its own set of competing

claims and demands, attests to the inherent difficulty of achieving social and political consensus on policy goals and programmed purporting

to serve the common interest.

‘Nk>hols, op. (’it., p. 7.

Because publics differ with each issue, no definition will be attempted here. It is assumed that

“the public” is demanding a greater role in decisions about science and technology, and that it

will continue to do so. The different publics that

coalesce around different issues vary widely in

their basic interests, their skills, and their

ultimate objectives. They are the groups that

will be heard in the widening debate about

scientific and technological issues, and are part

of what has been called the “social system of

science. ”9

The public has already become involved in

the decisionmaking process involving genetic

research. As the science develops, new issues in

which the public will demand involvement will

arise. The question is therefore: What is the

best way to involve the public in decisionmaking?

“J. Al. Zinl~ln, fublic K/u)w/@e (( ~iitlll)l’idgt~: (~ilt)~l)l.i(l~(l [ ll]i\wrsil.v PIWS, 1 !)68).

Issues and Options

Three issues are considered. The first is an

issue of process, concerning public involvement

in policymaking; the second is a technical issue;

and the third reflects the complexity of some

issues associated with genetics that may arise in

the future.

ISSUE: How should the public be involved in determining policy related to new applications of genetics?

The question as to whether the public should

be involved is no longer an issue. Groups demand to be involved when people feel that their

interests are threatened in ways that cannot be

resolved by representative democracy,

The more relevant questions are whether

current mechanisms are adequate to meet public desires to participate and whether a deliberate effort should be made to increase public knowledge. The last can only be accomplished by educating the public and increasing

its exposure both to the issues and to how people may be affected by different decisions.

OPTIONS:

A.

B.

Congress could specify that the opinion of the

public must be sought in formulating all major

policies concerning new applications of genetics, including decisions on funding of specific research projects. A “public participation

statement” could be mandated for all such

decisions.

Congress could maintain the status quo, allowing the public to participate only when it decides to do so on its own initiative.

If option A were followed, there would be no

cause for claiming that public involvement was

76-565 0 - 81 - 18
262 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

inadequate (as occurred after the first set of

Guidelines for Recombinant DNA Research

were promulgated). However, option A can be

implemented in two ways. In the first, the opportunity for public involvement is always provided, but need not be taken if there is no public

interest in the topic. In the second, public involvement is required. A requirement for public

involvement would pose the problem that if the

public does not wish to participate in a particular decision, then opinion will sometimes be

sought from an uninterested (and therefore

probably uninformed) public simply to meet the

requirement. Option A poses additional problems: What is a “major” policy? At what stage

would public involvement be required—only

when technological development and application are imminent or at the stage of basic

research? Finally, it should be noted that under

option A, if the public’s contribution significantly influences policy, the trend away from decisionmaking by elected representatives (representative democracy) and toward decisionmaking by the people directly (“participatory”

democracy) may be accelerated.

Option B would be less cumbersome and

would permit the establishment of ad hoc mechanisms when necessary. On the other hand, by

the time some issues are raised, strong vested

interests would already be in place. The growing role of single-issue advocates in U.S. politics,

and their skill in influencing citizens and policymakers, might abort certain scientific developments in the future.

Regardless of which option is selected, it

would be desirable to encourage different

forms of structuring public participation and to

evaluate the success of each method. Many different approaches to public participation have

been tried in the United States and Western

Europe in attempts to resolve conflicts over

science and technology. Some have worked better than others, but most have had rather

limited success.l0 Because public demands for

involvement are not likely to diminish, the best

1(J[ )i)l.[lll~v N[?l~ill iII)d h~ i[:hiie! Pollil[!k, “}’1’01)1(?111s iIINi 1%’(){:(?-

dUI’(W in I}W 11(’~iiiiitioll 01 ‘l’t’(sllllolo~ i(sill” Risk ,“ in SOCield Risk AsS~S.WTI~~I, Il. Schwing, illld W. ,III)(YIvJ ((+(1s.1 (N(?w Yot’k: Pl(!I~LII]]

1%’ss, 198(1).

ways to accommodate them need to be identified.

ISSUE: How can the level of public

knowledge concerning genetics

and its potential be raised?

If public involvement is expected, an informed public is clearly desirable. Increasing

the treatment of the subject, both within and

outside the traditional educational system, is the

only way to accomplish this.

Within the traditional educational system, at

least some educators feel that too little time is

spent on genetics. Some, such as members of

the Biological Sciences Curriculum Study Program, are considering increasing the share of

the curriculum devoted to genetics. Because

science and technology cause broad changes in

society, not only is a clearer perception of

genetics in particular needed, but more understanding of science in general. For about half

the U.S. population, high school biology is their

last science course. Educators must focus on

this course to increase public understanding of

science. Because students generally find people

more interesting than rats, and because human

genetics is a very popular topic in the high

school biology course, educators responsible for

the Biological Sciences Curriculum Study Program are considering increasing time spent on

its study in hopes of increasing public knowledge not only of genetics but of science in general.

At the university level, more funds could be

provided to develop courses on the relationships between science, technology, and society,

which could be designed both for students and

for the general public.

Several sources outside the traditional school

system already work to increase public understanding of science and the relationships between science and society. Among them are:

● Three programs developed by the National

Science Foundation to improve public

understanding of and involvement in science: Science for the Citizen; Public Understanding of Science; and Ethics and Values

in Science and Technology.
Ch. 13—Genetics and Society ● 263

●

●

●

●

Science Centers and similar projects specifically designed to present science information in an appealing fashion.

New magazines that offer science information to the lay reader-another indication

of increasing interest in science.

Television programs dealing with science

and technology. Examples are the two PBS

series, NOVA and Cosmos, and the BBC

series, Connections. CBS has also begun a

new series called The Universe.

Television programs dealing with social

issues and value conflicts. Particularly interesting is the concept behind The Baxters.

[n this half-hour prime time show, the network provides the first half of the show,

which is a dramatization of a family in conflict over a social or ethical issue. The second half of the show consists either of a discussion about what has been seen or of

comments from people who call in.

One interesting possibility would be to combine a series of Baxter-type episodes on genetic

issues with audience reaction using the QUBE

system, a two-way cable television system in

Columbus, Ohio (now expanding to other cities).

In this system, television viewers are provided

with a simple device that enables them to

answer questions asked over the television. A

computer tabulates the responses, which can

either be used by the studio or immediately

transmitted back to the audience. QUBE permits

its viewers to do comparison shopping in discount stores, take college courses at home, and

provide opinion to elected officials. It could be

effectively combined with a program like The

Baxters, to study social issues. If several such

programs on genetics were shown to QUBE subscribers, audience learning and interest could

be measured.

Any efforts to increase public understanding

should, of course, be combined with carefully

designed evaluation studies so that the effectiveness of the program can be assessed.

OPTIONS:

A. Programs could be developed to increase

public understanding of science and the rela -

tionships between science, technology, and

society.

Public understanding of science in today’s

world is essential, and there is concern about

the adequacy of the public’s knowledge.

B. Programs could be established to monitor the

level of public understanding of genetics and

of science in general and to determine whether

public concern with decisionmaking in science

and technology is increasing.

Selecting this option would indicate that

there is need for additional information, and

that Congress is interested in involving the public in developing science policy.

C. The copyright laws could be amended to permit schools to videotape television programs

for educational purposes.

Under current copyright law, videotaping

television programs as they are being broadcast

may infringe the rights of the program’s owner,

generally its producer. The legal status of such

tapes is presently the subject of litigation. As a

matter of policy, the Public Broadcasting Service negotiates, with the producers of the programs that it broadcasts, a limited right for

schools to tape the program for educational

uses. This permits a school to keep the tape for a

given period of time, most often one week, after

which it must be erased. otherwise, a school

must rent or purchase a copy of the videotape

from the owner.

In favor of this option, it should be noted that

many of the programs are made at least in part

with public funds. Removing the copyright constraint on schools would make these programs

more available for another public good, education. On the other hand, this option could have

significant economic consequences to the copyright owner, whose market is often limited to

education] institutions. An ad hoc committee of

producers, educators, broadcasters, and talent

unions is attempting to develop guidelines in

this area.
264 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

ISSUE: Should Congress begin preparing now to resolve issues that

have not yet aroused much public debate but that may in the

future?

As scientific understanding of genetics and

the ability to manipulate inherited characteristics develop, society may face some difficult

questions that could involve tradeoffs between

individual freedom and societal need. This will

be increasingly the case as genetic technologies

are applied to humans. Developments are occurring rapidly. Recombinant DNA technology

was developed in the 1970’s. In the spring of

1980, the first application of gene replacement

therapy in mammals succeeded. Resistance to

the toxic effect of methotrexate, a drug used in

cancer chemotherapy, was transferred to sensitive mice by substituting the gene for resistance for the sensitive gene in tissue-cultured

bone marrow cells obtained from the sensitive

mice. Transplanted back into the sensitive mice,

the bone marrow cells now conferred resistance to the drug.11 In the fall of 1980, the first

gene substitution in humans was attempted.l2

Although this study was restricted to nonhuman applications, many people assume from

the above and other examples that what can be

done with lower animals can be done with humans, and will be. Therefore, some action might

be taken to better prepare society for decisions

on the application of genetic technologies to

humans.

OPTIONS:

A. A commission could be established to identify

central issues, the probable time-frame for application of various genetic technologies to

humans, and the probable effects on society,

and to suggest courses of action. The commission might also consider the related area of

how participatory democracy might be combined with representative democracy in decisionmaking.

B. The life of the President's Commission-for the

Study of” Ethical Problems in Medicine and

Biomedical and Behavioral Research could be

extended for the purpose of addressing these

issues.

The n-member Commission was established

by Public Law 95-622 in November 1978 and

terminates on December 31, 1982. Its purpose is

to consider ethical and legal issues associated

with the protection of human subjects in research; the definition of death; and voluntary

testing, counseling, information, and education

programs for genetic diseases as well as any

other appropriate topics related to medicine

and to biomedical or behavioral research.

In July and September 1980, the Commission

considered how to respond to a statement from

the general secretaries of the National Council

of Churches, the Synagogue Council of America,

and the United States Catholic Conference that

the Federal Government should consider ethical

issues raised by genetic engineering. The request was prompted by the Supreme Court decision allowing patents on “new life forms. ” The

general secretaries stated that “no government

agency or committee is currently exercising

adequate oversight or control, nor addressing

the fundamental ethical questions (of genetic

engineering) in a major way, ” and asked that the

President “provide a way for representatives of

a broad spectrum of our society to consider

these matters and advise the government on its

necessary role, ”13

After testimony from various experts, the

Commission found that the Government is already exercising adequate oversight of the “biohazards” associated with rDNA research and industrial production. The Commission decided to

prepare a report identifying what are and are

not realistic problems. It will concentrate on the

ethical and social aspects of genetic technology

that are most relevant to medicine and biomedical research.

The Commission could be asked to study the

areas it identifies and to broaden its coverage to

1‘J(?iill [,. hlil]s~, “(kilt? ‘l’lsitl]sf(?l’ (;h’en ii New Twist,” Scienc e

~()~:~~, ,\pril 1980, p. 386.

‘2(;i!]il l\iii’i Kolii[ii illl[{ Ni($holiis Wit(It?, “t{tlt]liiil (kIM ‘l’ls~iill]l~t~t

Stirs N(IW I)(II);I[(I, ” Science 2 10:24, octolwr 1980, p. 407.

t~slillell)ellt I)v the gellel.il] s~t:l’~t~l.i~s, (1 .S. ‘C:alholic [k)ll~t?l’-

(vM:[?, or”igi[ls, N( I l)()(!tltllelltiilm~ Service, vol. lo, N(.). 7, JLII.V Li,

1 !)/J().
Ch. 13—Genetics and Society . 265

additional areas. This would require that its

term be extended and that additional funds be

appropriated. The Commission operated on $1.2

million for 9 months of fiscal year 1980 and $1.5

million for fiscal year 1981. Given the complexity of the issues involved, the adequacy of this

level of funding should be reviewed if additional

tasks are undertaken.

A potential disadvantage of using the existing

Commission to address societal issues associated

with genetic engineering is that a number of

issues already exist and more are likely to appear in the years ahead. Yet there are also other

issues in medicine and biomedical and behavioral research not associated with genetic

engineering that need review. Whether all

these issues can be addressed by one Commission should be considered. There are obvious

advantages and disadvantages to two Commissions, one for genetic engineering and one for

other issues associated with medicine and biomedical and behavioral research. Comments

from the existing Commission would assist in

reaching a decision on the most appropriate

course of action.

Bibliography: suggested further reading

Dobzhansky, Theodosium, Genetic Diversity and Human Equality (New York: Basic Tools, 1973).

A discussion of conflicts between the findings

of science and democratic social goals. Detailed

coverage of the scientific basis for present debates about intelligence and the misconceptions

often involved in genetic v. environmental determinants of certain human traits.

Francoeur, Robert T., “We Can - We Must: Reflections on the Technological lmperative, ” Theobgical studies 33 (#3): 428-439, 1972,

Argues that man is a creator by virtue of his

special position in nature, and that humans must

participate in deciding the course of their evolution.

Goodfield, June, Plaving God: Genetic Engineering and

the Manipulation of Life (New York: Harper Colophon Books, 1977).

Discusses the benefits, problems and potential

of genetic engineering. Describes the moral

dilemmas posed by the new technology. Suggests

that the “social contract” between science and

society is being “renegotiated.

Harmon, Willis, An Incomplete Guide to the Future

(New York: Simon and Schuster, 1976).

Surveys how social attitudes and values have

changed throughout history and how they may

be changing today. Argues that mankind is in the

midst of a transition to new values that will affect

our world view as profoundly as did the industrial revolution in the 19th century.

Holton, Gerald, and William A. Blanpeid (eds.), Science and Its Public: The Changing Relationship

(Boston: D. Reidel, 1976).

A collection of essays on the way science and

the society of which it is a part interact, and how

that interaction may be changing.

Hutton, Richard, Bio-Revolution: DNA and the Ethics

of Man-Made Life (New York: New American Library (Mentor), 1978).

Reviews the history of the debate about recombinant DNA, discusses the scientific basis for the

new technologies, and discusses the changing

relationship between science and society. Suggests how the controversies might be resolved.

Monod, Jacques, Chance and Necessity (New York:

Alfred Knopf, 1971).

A philosophical essay on biology. Two seemingly contradictor-v laws of science, the constancy of inheritance (“necessity”) and spontaneous

mutation (“chance”) are compared with more

vitalistic and deontological views of the universe.

An affirmation of scientific knowledge as the only “truth” available to man.

Nichols, K. Guild, Technology on Trial: Public Participation in Decision-Making Related to Science and

Technology (Paris: Organization for Economic Cooperation and Development, 1971).

Reviews mechanisms that have been used by

countries in Europe and North America to settle

disputes involving science and technology.
266 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Sinsheimer, Robert, “Two Lectures on Recombinant

DNA Research,” in The Recombinant DNA Debate,

D. Jackson and Stephen Stich (eds.) (Englewood

Cliffs, N. J.: Prentice Hall, 1979), pp. 85-99.

Argues for proceeding slowly and thoughtfully

with genetic engineering, for it potentially has

far-reaching consequences.

Tribe, Laurence, “Technology Assessment and the

Fourth Discontinuity: The Limits of Instrumental

Rationality,” Southern California Law Review 46

(#~): 617-660, June 1973.

An essay on the fundamental task facing mankind in, the late 20th century: the problem of

choice of tools. New knowledge, especially from

biology, will increasingly offer options for technology, the use of which will cause changes in

human values.
Appendixes

I-A. A Case Study of Acetaminophen Production . . . , . . . . . . . . . . . . . . . . . . . .269

I-B. A Timetable for the Commercial Production of Compounds Using

Genetically Engineered Micro-Organisms in Biotechnology . . . . . . . . . . . .275

I-C. Chemical and Biological Processes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 292

I-D. The Impact of Genetics on Ethanol—A Case Study . . . . . . . . . . . , . . . . . .. 293

II-A. A Case Study of Wheat. , . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .304

II-B. Genetics and the Forest Products Industry Case Study. . . . . . . . , . ......307

H-C. Animal Fertilization Technologies. . . . . . . . . . . . . . . . . . . . . . . . , . . . . . . .309

III-A. History of the Recombinant DNA Debate . . . . . . . . . . . . . . . . . . . . .. ...315

HI-B. Constitutional Constraints on Regulation , . . . . . . . . . . . . . . . . . . , . . . . . .320

III-C. Information on International Guidelines for Recombinant DNA . . . . . , .. 322

IV. Planning workshop Participants, Other Contractors and Contributors, and

Acknowledgments. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . , , .329
——

Appendix I-A

A Case Study of

Acetaminophen Production

Summary

‘1’he ol]jeclil’e ot’ Ihis case slud~ is 10 denmnstriite

the [?cononlic t’t?asihilit.v of applying a genetically

engineere{i strain to n]ake a chemical product not

tlmv produced bv t’t?i’lll[;ilt:itio[l.

.

E3ACKGR0UNIl

tl(;[?tiit]~it~()~)ll[~tl (~\P~]II) M’as chosen for the cas e

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on the market. Around 20 million pounds (lb) are

manufactured annually. Mallinckrodt, Inc., produces

60 to 70 percent; the remainder is llliillllf’:l{: tlll’e(~ primaril~~ b-v (;l](~ Itltt?l>lliltioll:iliitld Nlo]lsiinto” (1). A PAP

is sold to l~(?i~ltl~ [;iil’[? (;ol~~l]iitlies, ~i’hich [~~iit’k(?t it to

ret a il(?rs.

‘1’he hfcNeil (k)IlstItl]er Products di~’ision of Johnson &, Johnson, itrhich ll]ill’kt~ts APAP Lllldel’ the

tt.ii(!e IIiIIIM?, ‘J~vl(?IIol, h{is th[~ l:it’~~st shaI’~ of th e

market. (her a dozetl other con~panies in the United

Stilt[?S S[?11 it Lllld(?l’ othel” tl’;id(? Iliill](?!j.

one ch[?n] ical l~]ittlllf~i(;tllt’(?l”s bulk selling price for

,lp,]p is ;,~-{~~~~~{i $~G~~/]l),l BV th(? tin]e the consumer

f)l]l’{:hiis[~s it ilt the [Itsug st[)i’e, the markup results in

ii selling price ot” ill’(llill(i $25 to $50/ltl, (iepen[iing 011

[iosagf? anci package sixes. ‘1’hus, the? total t’alue of

,\Ilt~p to t}l(? ll];lllllf~l(:tlil.f?s k S0111[! $~o Il]i]]bn annu”

illl~, while tilt? totill retail \’alU{? fallS in th(? 1’iill#? O f

$500” million to $1 billion.

●

●

●

●

APPROACH ES

A (:olls[;l’~’;lti~~[? approach }i’as taken, in that onley a

(}otl\’[?Iltiotl;il” batch fermentation process was consi(iermi.

\’arial)les were selectwi i]ertaining to the choice of

the microhial pathway; the nature of the feedstock; con~rersion ei”ficiencies of feedstock t o

~1 PA P; anci the finiii yielci of APAP.

(;osts were basefi on proprietary processes involving startup, large-scale fermentation, and recovery

of /1 PA P.

(k)sts were itemized for materials and supplies;

li~t)or tiistributi on; utilities (hroken down by specific energy l’e(ll]il’[~t~]t;tltsi~(;~ot’ciitl~ to process and

equipment ); equipment (groupe[i according to

●

●

●

●

process); and builciing l’e(l~lil’t~ll~t~l~ts (S]}iiCt? nee[is

allocate~i accorfi i ng 10 process).

CONCLUSIONS

‘1’he projected cost for illiiilllf~i(:tll]’iflg APAP b-v

means of hatch t“(;l”ttl(;llt[itioll,using a genetically

engineered strain, amounts to $ 1.OS/111. This cost is

based on a plant pro(iucing 10 million 11] of APAP

iill Iltliliicv.

A S a I}llle (It’ tht]l~ll), tht? ~1’OSS tl)iirgitl ~01’ lllal~l]fa~-

t~ll’e Of ii (:hellli[:iil SIICI1 ;1S /11]111) ShOUld aJ)Pl’OXimate 50 percent of SiiieS. ‘1’}1[? gross margin represents the profit before general ami ilcifllitlistl’;iti~’e,

marketing an~i selling, anci research an[i (development expenses. ‘I”IIC ~1’OSS margin f“ol” iill of the

prd~]cts miide by hliillii~(:ki’()[it, th(? largest illii[lllfilCt U1’el’ of l~~~~p, iilllollfltd to ~~ iiild 37 p(?l’Cellt

of Siil~S in 1977 itll(i 1978, resfmctil’el. v.z The gross

lllill’~ill fol” NIOilSilIlto, ii lllLICh liil’~t?l’ Colllpiilly

thiin hfiillinckrodt I)llt it sll~iill[?l’ il]iltll]fii~tlll’~1’ of

,][JAI}, ill]l{)~lllt(?~i [() 27 illl[l 26 })(?l’C(~llt (If all SaJeS

in 1976 illl(i 1977, l’t?S~)eCt i\wlv. s It’ ttl(? @’OSS llliil’-

gin f[)r APAP is iis hi@ iis 50- p~?r[:ent of si~les, its

Clll’1’ellt COSt Of lllilllllfiI(lt Llf’[? ShOUlti iilllOLlllt tO

$1.325/11), based On ii bltlk selling price of $2.65/111.

Therefore, its projectmi cost ~then pr(xiuced l~y

t“ernlentiition is around 20 percent I(n%’er than its

estimateci cost Whetl pro(iuce(i b-v Cht:llliCiil s.Vnthesis.

If the selling price of APAP protiuce~i hy fel’llletltiition is l~]iil.kd LIp 100 percent, the bulk selling

price becomes $2.10/ib. This decrease of $0.55/lb

could be transformed into cost savings of around

$5 to $10/lb to the consumer. These economies

would result in an annual cost saving to the consumer of $100 million to $200 million.

(hrrent processes for synthesizing A PAP from

nitrobenzene do not ii~pei~r to pose significant

pollution problems, iilthOU~h ii number Of side

products are formed anti must he renwved.4 5 G

Howe\~er, ii fermentiition process would be even

1 Klilll i 11(’A 1’()(11, I IN’,, ,4/11111;1/ /{ f’/)(Wl, I !)7M.

‘Jll)ll%illllo ( ‘(). , ,,ltl/)t/;l/ Ii f’ixwl, 1 !)77.

it{, (;, 1](,1,11(,111 11),.()(,(IS5 [~)1, ))1(,1~:11.illg ,\illilloj~llf)tlol,”” 1 ‘..S. P;ll(’111

:i,:itKi,4 16, I !)68.

‘l:. J\. I!ill’oil, 1{. (;. Ii(’11111’l’, illl(l \. l’,. \\’(’illl)(’l.g, “1’111’il’i(’ill iotl of

/)-,\lllitlol)ll(ltlol,” “ ( 1,,$. p;~/(,/1( :],(;:)4 ,3(18, I !)72.

“l”. \. I}ill’oll itll(l R. ( ; Ik’1111(’l’, ,11(11.if’i(.;l[iol~ (]1’ /).l\ll]illol)ll(,llol,””

( ‘.s. }’;llrlll :},7 I 7![;so, I !)7:}.

269
270 “ Impacts otApplied Genetics —Micro-Organisms, Plants, and Animals

(~]eiiner. (111].v A PAP W(IU]d iiccL]n~uliit~: ill] ~thel’

llM?tiit)Olitt?S il[’(? lliitlll’illl.’ occurring. Even nlicro-

()]’~iitlit+nls COLIM be c[dl~(:t(d itft~r ~iich biitch iind

I)IYXXSSCX1 into ii ciik~ tot” US~ iis ii high protein

illl illlill feed.

Biological parameters

MICROBIAL PATHWAY

A proposed ~iithwii~ for convertin~ aniline to

A PAP viii the il~et~liiti(}n of iin intermediilte, p-aminophend, is shown in figure l-A-l. !’iiI’iOLIS fungi have

ken identified in which these reiictions occur.7 s 9

7R. \’. Smith atld J. P. Rosii~~a, “hlk]sobiiil Models of Miill~nli{liiitl

hfetiibolisnl, ’” J. Pharmarxw. Sci. 64: 1737 -17s9, 197s.

‘R. t’. Smith iilld J. P. liosit~~a, “M icrol)ial Models of Mammalian

hl(?tiil)olisn): Aron)iilk: H.vtit.l]xvlatiotl, ” Arch. Biochern. Biol]hvs.

16 1:551-.5.58, 1974.

. .

‘\’. R. klunzner, It. hlutschlwl iind h!. Rummel, “Llher die n)ikroI)iologische unwandlung N-hitltiger substrate” K:oncerrling the

Microbiological “1’1’utlslol’nlatiotl of N- containing Substrate), Plant

Medica 1 s:97- i03, I Y67.

Figure l-A-l. Bioconversion of Aniline to APAPa

NHC10CH3

aceticacid?f I

/

.

N H2

I

N H2

/

Ho

I

p-aminophenoi

Aniline

t~ltertliitiv~l.v, ilniline could he ii~et.yliited directl y

forming i[cetiit~ilide, which in turn would be hydrox-

~liit~d to A PAP. ‘“ ‘‘ 12 A l~un]her of Streptomyces species hiiv~ heen f(}u[ld to cotlvei’t iicetitnilide t o

APAP.13 “1’he ~iithwii~ hnrolvhlg p-iiminophenol wa s

chosen simply be(:iiuse the conversion efficiency of

it~et ic ii~id to A PAP would l)e slightly higher if acetic

iicid entered the ~\’erilll l’~il~t ion i]t the second step

[’iith~[. thii[l iit the first step.

HOST MICRO-ORGANISMS

The most suiti~hle ltlicl>()-()l.gi~tlisn~ for production

of APAP in ]iir~e-s(~iil~ fermentiition may not necesSill’iluV he one thilt 1101’ llliill~ nletilbolizes aniline 01’

~)-itl~~il~~~llet~ol. While ii hycterium might serve as ii

Sllitiit)k host for insertion iitld expression of the recluisite genes, it yeiist il~ii~ represent ii hetter choice. [t

will Prot]iit)l.v more closeltv resemble the organism

[IWIN which the genes are isoliited.

Fermentation efficiencies

CONVERSION EFFICIENCIES

The n]olilr iil]d weight conversion efficiencies for

the hioconversion of feedstock to product are projected in tithle 1-A-1. ‘l’he bioconversion of aniline to

Iuslllit h, et al., op. d.

I 1 hfun~n~r, [?t ill., op. rit .

“R. J. ‘1’heriiiult iil~d ‘l’. }1. l,ongtieki, “hlicrohial Conversion of

Acetiinilide to 2’-llydl’oxyii(:etal~i]ide and 4’- Hydl.oxya(:etiil] ili[ie, ”

Ap/. ,\ficrobio/. 15: 1431-1436, 1967.

131hid.

Table I=A-l.—Fermentation Efficiencies to Meet the

Requirements for the Production of Acetaminophen

(APAP) From Aniline

Overali molar conversion efficiency of:

(a) Aniline to APAP. . . . . . . . . . . . . . . . . .

(b) Acetic Acid to APAP. . . . . . . . . . . . . .

Overali weight conversion efficiency of:

(a) Aniline to APAPa . . . . . . . . . . . . . . . . .

(b) Acetic Acid to APAPa . . . . . . . . . . . . .

Utilization of:

(a) Aniline in fermentation broth . . . . . .

(b) Acetic acid in fermentation broth . . .

Production of APAP in broth . . . . . . . . . .

Batch volume . . . . . . . . . . . . . . . . . . . . . . .

Recovery efficiency. . . . . . . . . . . . . . . . . .

Yield of APAPlbatch . . . . . . . . . . . . . . . . .

Number of batches/year . . . . . . . . . . . . . .

Annuat yield of product. . . . . . . . . . . . . . .

90.25Y0

95.0

146.5

239.1

2.28 lb/gal

1 .39)gal

3.34 lb/gal

33,500 gal

90.0 %

100,701 lb

100

10,070,100 lb

aApAp = Npacetyl.p.amlnophenol = acetaminophen = p.acetamidophenoi = Phydroxyacetanilide =Tylenol (trade name of McNeii Laboratories).

SOURCE: Genex Corp.

%erait weight conversion of precursor to APAP =

molecuiar weight of APAP x molar conversion efficiency

molecular weight of precursor of precursor to APAP

SOURCE: Genex Corp.
Appendix l-A—A Case Study of Acetaminophen Production ● 271

~\f}/\ P inlwh’cs two steps. ‘1’he product of the inciividII;II I’t?il(;t ions f’ol” [?it(Yll St(?j) l’(?pl’E!St?lltS the ()\’(?I’iill

con\wrsion (? fficit?ncv. tl Illolill” conversion efficiency .

of !)5 l)eITCIII \t’iis iisslltll[![l tot” ~ii(;h step. This i~iill]~

is l)ilS(?({ 011 ii nlultitud[? of l’t?pOl’t S Clelll(lllStl’ilt ill~

Sillliliil’ Illolill ” colli’crsion efficiencies tot” alliilO~OLIS

I)io(;ht?llli(:ill l’(?it(:tiollS lllld[?l’ ii(~tllill f“(?l’lll~ll tiltioll

conditions. I 4

PRODUCT YIELD

‘1’he yield of /\P/lP projected in tiihl~ l-A-l is I]iisd

011 t;st iil]iitill~ ii rittio of J() pCIYWIIt weight to VOIUtI]~

(i.e., 4(1 II) pt?t’ 10(1 gilllollS (g:Il) Of teI’lllelltiitiO[l bl’Oth)

I)rior to 90 pcr(x?tll r(?(xn(?r~ . Such ii high yield is pern]illcd })[?(;iills[? ()[” th(! poor” soluhi]itv of APAP under

()~)(?t’iit ing condi[ ions. /\s ii IWSLI]I, h@ IC\r~]S of APAP

\\’olll[l llil\’[! 110 il[l\’(?l’S(? t?f’t’(?(;t 011 tht? hOSt llliCl’O-Ol’gilnism. [ lse of insoilihlc s.vstcms in f“f~t’ill[?lltati(~ll hilS in

fact been reported in recent years—e.g., in certain

microbial transformations of steroids, yields of 40

percent may result due to the insolubility of the

product.

III{ J il)l)O II, ‘I I I II II 11)1)11 Izlt(l((lll s,’” ill ,\/i/l{/;~//{fJ/~(jrl,s (m/.’rrlll(vl-

/;i/i(~/t /)l’(l(’(’<s,s( ’.$, \(ll 1, 1) 1)(’t”llllall (I’(l) LY(}lt }ol’h: \(’il(l(’llli(’

l,l!,,~>, 1!)77), ])]) zo.T-2:l:\

Table l-A-2.—Summary of Production Conditions

of APAP

Economics

PROD1JCTION RE[~lJIREMENTS

liow the tiiriolts production t’[?c~llil’[?tlletlts would

I)t? met during th(? Inicl’ohiill tl’illlSfol’lllilti oil” of” illliIine to tlPAP is SLlllllllill’iZ[?d ill tiihlt?s I-A-1 iin(~ 2. AniIin(? ittld ii(:(?ti{: ilci[i WOUl(i not i)[? it(i(id to tilt? f[?l’-

Illt?lltiltioll 1)1’ottl iiii ill ollC(? t)llt l’iitil~l- step-wis[? il(Y-

(x)r(i ing to til(?ir i’iit~s of” coniwrsion. ‘1’he pi;~nt ivoui(i

(X)lltilill tlt’() 50, ()()( )-~ili f(?l’lll(?ll t[?l’S, i%’hictl in th[?

(:0111’S[? of il .Vf?iil’ W’olli(i .vi[?i{i 1() n~iliion lb Of t\ PAP.

PRODUCTION COSTS

‘1’ilt? COStS fO1’ ttl[? illllllliil ~1’OCiLICtiOll iI1’~ SLllllllliiriz(?ci in till)i~ I-A-3. ‘1’he.v iit’[? I)rok[:t) dO\VIl into their

Illiij(ll’ COllll)oll[+lltS iill{i iil’e (? Xpl’[?SSf3[i i)otil ii!$ iillllllill

COStS iilld ii!+ unit COStS. Ik?tiiil[?(i tlUCi#?tS t[)l’ tht? t’ill’i011S COSt (X?llteI’S ill’f? ShOWll in tillll~S 1-A-4 till’ oll~il

l-A-lo. Miiteriiils iiild suppiies iir~ describe(i in tiil)l(?

l-A-4; iilhol” [iistrihut ion in tiil)]~ l-A-5; utility requir (?-

Il?ellts in till)l[?S l-tl-~ tilrOu~h I-t\-/l; equipnlent ill

till)le l-/~-9; i~lld sj)ilc~ Isf;(ll]ii>[?ll][+flts in tiil)l~ I-A-IO.

‘1’ilis iiiliil~sis l’[?~~~iils ii unit (;ost Of APAP [?({iiiii to

$1 .(wit).

Table 1. A.4.—Materials and Supplies for

Production of APAP

Number of fermenters . . . . . . . 2

Size of fermenters. . . . . . . . . . . 50,000 gal

Operating volume . . . . . . . . . . . 33,500 gal

Cycle. . . . . . . . . . . . . . . . . . . . . . 7a

Batches . . . . . . . . . . . . . . . . . . . 100

%-day fermentation, l-day turn around.

SOURGE: Genex Corp.

Materials Cost/batch Cost/year

Fermentation

Fishmeal (l.5% @ $0.155) . . . . . . . $ 648.68 $ 64,868

Glucose (1.5% @ $0.1535). . . . . . . 642.40 64,240

Lard oil (2.5% @ $0.325) . . . . . . . . 2,266.88 226,888

Mineral salts (4,215 lb @ $0.05074) 213.77 21,377

Aniline (76,250 lb@ $0.42). . . . . . . 32,027.52 3,202,752

Acetic acid (46,680 lb@ $0.245) . . 11,436.60 1,143,660

Miscellaneous (1OYO of basic

materials). . . . . . . . . . . . . . . . . . . 377.17 37,717

Subtotal. . . . . . . . . . . . . . . . . . . . $47,613.02 $4,761,302

Table l-A.3.—Summary of Costs of

Production of APAP

Annual cost Costllb

Materials and supplies . . . . . . . $ 6,133,802 $0.6091

Labor. . . . . . . . . . . . . . . . . . . . . . 2,012,140 0.1998

Utilities. . . . . . . . . . . . . . . . . . . . 630,200 0.0626

Equipment . . . . . . . . . . . . . . . . . 1,377,590 0.1368

Building . . . . . . . . . . . . . . . . . . . 439,399 0.0436

Total . . . . . . . . . . . . . . . . . . . . $10,593,131 $1.0511b

Annual production = 10,070,100 lb

Recovery

Filter aid(0.21b/gal @ $13). . . . . . . $ 871.00 $ 87,1OO

Other chemicals and supplies. . . . 1,600.00 $ 160,000

Subtotal . . . . . . . . . . . . . . . . . . . . $2,471.00 $ 247,100

Finishing

Packaging (1,255 bag units

at$0.80) . . . . . . . . . . . . . . . . . . . . $ 1,004.00 $ 100,400

Other (labels, stencils, etc.). . . . . . 1,004.00 $ 100,400

Subtotal . . . . . . . . . . . . . . . . . . . . $2,008.00 $ 200,800

General supplies

Maintenance (4°Aof capital investment) . . . . . . $ 425,900

Other (laboratory office, plant miscellaneous) . 498,700

Total . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . $6,133,802

SOURCE: Genex Corp SOURCE: Genex Corp.
272 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Table l-A-5.-Labor Distribution for Production of APAP

Salary and wage cost

Category Man-hours per week Hourly rate $/week $/year

Supervision

General manager . . . . . . . . . . . . . . . . . . . . . . . . .

Superintendents. . . . . . . . . . . . . . . . . . . . . . . . . .

Managers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Supervisors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Hourly rated employees, services

Laboratory

Level I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level II. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level Ill . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level lV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Maintenance and engineering

Level l . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level lI. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level lIl . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level IV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Hourly rated employees, production

Fermentation department

Level l . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level II . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level Ill . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level lV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Recovery department

Level l . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level II . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level Ill . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Level IV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Subtotal. ......, . . . . . . . . . . . . . . . . . . . . . . . . .

40

80

80

320

80

80

120

40

240

240

240

160

200

240

80

80

320

400

80

120

. . . . . . . .

20

17

15

12

10

8

6

5

10

8

6

5

10

8

6

5

10

8

6

5

...,. . . . .

$ 800

1,360

1,200

3,840

800

640

720

200

2,400

1,920

1,440

800

2,000

1,920

480

400

3,200

3,200

480

600

. . . . . . . . . . . . . . . . . . . . .

Add overtime@ 6% x 1.5 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Subtotal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Add fringe benefits @25%. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Total salaries and wages. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

$ 41,600

70,720

62,400

199,680

41,600

33,280

37.440

10,400

124,800

99,840

74,880

41,600

104,000

99,840

24,960

20,800

166,400

166,400

24,960

31,200

$1,476,800

132,912

$1,609,712

402,428

$2,012,140

SOURCE:GenexCorp.

Table l-A-6.-Steam Requirements for

Production of APAP

Operation Lb/batch

Sterilization, fermenters, and seed tanks:

Heating . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52,100

Holding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20,000

Sterilization, piping, and equipment (other) . . . 20,000

Heating acetaminophen solution (recovery) ... 163,500

Drying, turbo dryer. . ......................200,300

General purpose usage . . . . . . . . . . . . . . . . . . . . 50,000

Total. . . . . . . . . . . . ......................505,900

Cost at $5.00/Mlb:

Per fermenter batch =$ 2,530

Per year (100 batches) =$253,000

SOURCE:GenexCorp.
Appendix l-A—A Case Study of Acetaminophen Production . 273

Table l-A-7.–Electricity Requirements for Production of APAP

Units/batch

Connected load HP kW (hours operation) kWh

Fermenters. . . . . . . . . . . . . . . . . . . . . . ........200

Seed tanks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47.5

Chillers . . . . . . . . . . . . . . . . . ................580

Air compressor . .........................275

Harvest tank.... . ........................10 0

Decanter centrifuge. . .....................120

Process tanks. . . .........................300

Crystallizing tanks . .......................300

Turbo dryer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

Cooling tower.. . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Pumps (est. =6@ 7.5) . . . . . . . . . . . . . . . . . . . . 45

Lighting, instruments and general load. . . . . . . 25

Total kWh . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

@O.05/kWh = $3,469 per batch

@100 batches/yr= $346,900 per year

149

35

433

205

90

224

224

22

30

34

19

144

24

86

11

52

19

23

144

144

144

21,456

840

4,763

17,630

825

4,680

4,256

2,464

4,320

4,896

2,736

69,372

SOURCE:GenexCorp.

Table I-A.8.-WaterRequirements for

Production of APAP

Gal/batch

Fermentation . . . . . . . . . . . . . . . . . . . . . . . . . . 35,000

Tower makeup . . . . . . . . . . . . . . . . . . . . . . . . . 63,000

Process loss . . . . . . . . . . . . . . . . . . . . . . . . . . 100,000

Chilled water makeup. . . . . . . . . . . . . . . . . . . 30,000

Direct cooling . . . . . . . . . . . . . . . . . . . . . . . . . 50,000

General use..... . . . . . . . . . . . . . . . . . . . . . . 25,000

Total . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303,000 gal

Process Watergate =$1.00/M gals

cost = $303/batch

100 batches/yr =$30,300/year

SOURCE:GenexCorp.
274 ● Impacts of Applied Genetics —Micro-Organisms, Plants, and Animals

Table l-A-9.—Equipment Costs for

Production of APAP

Receiving and batching area

320,000 gal steel aniline storage tanks,

insulated and cooled - @ $47,000 . . . . . . . . . . . $ 341,000

220,000 gal aluminum acetic acid storage

tanks, insulated and cooled - @ $71,300 . . . . . .

110,000 gal steel nitrogen storage tank with

controls and instruments. . . . . . . . . . . . . . . . . .

110,000 gal steel lard oil storage tank,

insulated and heated . . . . . . . . . . . . . . . . . . . . .

110,000 gal stainless steel Batch tank with

programmable controller and agitator. . . . . . . . .

21,700 ft3 stainless steel Hopper bins with

conveyors - @ $58,100 . . . . . . . . . . . . . . . . . . . .

1 Electric forklift truck . . . . . . . . . . . . . . . . . . . . . .

Fermentation and seed area

1150 gal stainless steel seed vessel, fully

instrumented. . . . . . . . . . . . . . . . . . . . . . . . . . . .

12,500 gal stainless steel seed vessel, fully

instrumented. . . . . . . . . . . . . . . . . . . . . . . . . . . .

250,000 gal stainless steel fermenters, fully

instrumented with central control room -

@ $399,000 . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Recovery area

150,000 gal stainless steel process tank,

cooled, agitated and insulated . . . . . . . . . . . . .

13,000 gal steel filter aid slurry tank

with agitator . . . . . . . . . . . . . . . . . . . . . . . . . . . .

1 Stainless steel continuous decanter

centrifuge . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

2100,000 gal stainless steel process tanks,

insulated with external steam injection heater,

pump and agitator- @ $333,000 . . . . . . . . . . . .

342,600

47,000

22,300

59,500

116,200

11,400

125,000

169,000

798,000

195,000

11,300

167,000

666,000

120,000 gal stainless steel side-entering surge

tank with agitator . . . . . . . . . . . . . . . . . . . . . . . . $ 56,100

350,000 gal stainless steel crystallizing tanks,

insulated with heavy duty cooling coils and

top-mounted agitator - @ $195,000 . . . . . . . . . .

1 Stainless steel turbo tray dryer. . . . . . . . . . . . . .

23,500 ft3 stainless steel hopper bins-

@ $66,000 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

1 Bagging unit. . . . . . . . . . . . . . . . . . . . . . . . . . . . .

4 Stainless steel finished product conveyors-

@ $12,000 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Auxiliary equipment

31,500 c.f.m. reciprocating air compressors -

@ $168,000 . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Laboratory and office equipment , . . . . . . . . . . . .

Chillers, 500 ton total capacity . . . . . . . . . . . . . . .

1 Cooling tower, 1,500 g.p.m.. . . . . . . . . . . . . . . . .

35 Pumps and motors, various sizes. . . . . . . . . . .

2 Dump trucks -$12,000 . . . . . . . . . . . . . . . . . . . . .

Ventilation, general and spot - @ 7.5%

of equipment. . . . . . . . . . . . . . . . . . . . . . . . . . . .

Piping, general, materials and installation -

@ 7.5% of equipment. . . . . . . . . . . . . . . . . . . . .

Miscellaneous equipment (hand tools, etc.) -

@ 5°A of equipment . . . . . . . . . . . . . . . . . . . . . .

585,000

653,000

132,000

20,000

48,000

498,000

650,000

575,000

210,000

104,700

24,000

583,791

583,790

389,194

Total . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . $7,783,875

Annual charge for capital recovery over 10-year

period, with 12% interest compounded

annually ($7,783,875 x 0.17698) . . . . . . . . . . . . $1,377,590

SOURCE: Genex Corn.

Table I-A-10.–Buiiding Requirements for Production of APAP

Area Gross space ft2

/ft3 Unit valuea

cost

Central office . . . . . . . . . . . . . . . . . . . . 940 41.00 b

$ 38,540

Laboratories. . . . . . . . . . . . . . . . . . . . . 4,500 70.00b

315,000

Warehouse . . . . . . . . . . . . . . . . . . . . . . 2,000/36,000 27.00b

54,000

Batching . . . . . . . . . . . . . . . . . . . . . . . . 1,000/30,000 1.75 52,500

Fermentation (including seed) . . . . . . 6,000/320,000 1.75 560,000

Harvest, filter . . . . . . . . . . . . . . . . . . . . 3,500/169,000 1.75 295,750

Processing, crystallization . . . . . . . . . 8,700/470,000 1.75 822,500

Drying, finishing. . . . . . . . . . . . . . . . . . 5,000/270,000 1.75 472,500

Warehouse, finished product. . . . . . . 11,000/200,000 27.00b

297,000

Auxiliary equipment. . . . . . . . . . . . . . . 4,300/154,000 1.75 269,500

Maintenance, engineering. . . . . . . . . . 11,500/207,000 1.75 362,250

Total. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . $3,539,540

Amortization over 30 years@ 12% compound interest $439,399C

aunit values in cubic feet except where noted by “b.”

bunit value in square feet.

cAmortizatlon = 0.12414 X tOtal.

SOURCE: Genex Corp.
Appendix I-B

A Timetable for the Commercial

Production of Compounds Using

Genetically Engineered MicroOrganisms in Biotechnology

Objectives

● The estimation of the proportions of various

groups of commercial products and processes f o r

which recombinant DNA (rDNA) technology could

be applicable.

● The construction of timetables to indicate plausible sequenccs of commercial developments that

would result from the application of rDNA technology.

Approaches

The following five industries were evaluated:

1. pharmaceutical,

2. agricultural,

3. food,

4. chemical, and

5. energy .

The manufacturing processes that would result

from the application of rDNA technology would be

based on fermentation technology, Therefore, a set

of parameters was developed to serve as a guide to

assess the economics of applying fermentation technology to the manufacture of products currently

manufactured by other means.

The chemical industry generates a large number

of products that could be attributed to (and is in this

study) the other four industries cited, this particular

industry was focused on more closely than the

others. The following factors were considered in

constructing the timetables showing the applicability

of rDNA technoloGY:

●

●

●

●

●

the current state of the art of genetic engineering;

the current economic limitations of fermentat ion technology;

the length of time to progress from a laboratory

process to the pilot plant to large-scale production;

the plant construction time; and

the Government regulatory agency approval re -

quired (of the products and manufacturing

processes, not of the rDNA technology per se).

Sources of information

While much of the information compiled for this

report was obtained from published sources, a considerable amount came from prior proprietary studies performed by Genex Corp. In the latter case, information is used that is not proprietary, although

the sources must remain confidential. In this connection Genex has had numerous discussions with the

technical and corporate management of more than

100 large companies (generally multibillion dollar

companies), concerning research interests, product

lines, and market trends. Production costs are extrapolated for four fermentation plants of various

sizes and capabilities. (See table I-B-1.)

A group of Genex scientists, consisting of a biochemical engineer, two organic chemists, a biochemist, and four molecular geneticists rated the feasibility of devising micro-organisms to produce various

chemicals in accordance with the fermentation conditions specified in table I-B-1. For those chemicls

that appeared to be capable of being produced microbiologically, dates were assigned for the times

when the necessary technology would be achieved in

the laboratory. By combining both technical and economic factors, it then became possible to project a

timetable for commercial production. (See table

I-B-2.)

It should be emphasized that an extremely conservative approach was taken in considering fermentation economics over the next 10 years. only the relatively poor economics of conventional batch fermentation was considered. immobilized cell processes were projected to be 15 years away, and even

then, the incremental cost savings projected (see

table I-B-1) are lower than the incremental cost savings currently obtained with immobilized cell processes. The assumptions made here, however, did include reasonably high product yields and highly effi275
276 ● Impacts of Applied Genetics —Micro-Organisms, Plants, and Animals

Table I-B= I.-Unit Cost Assumptions for the Production of Chemicals by

Fermentation After Various Intervals of Time

Annual ca Unit cost

excluding excluding Complete

Earliest date Size of plant Type of Product yield precursor precursor unit cost

(year) (lb) fermentation (%) ($ millions) ($/lb) Precursor (W/lb)

5 50 Ordinary batch 12 23.5 0.47 Petrochemical 0.66

10 Ordinary batch 40 24.5 0.25 Petrochemical 0.44

200 Immobilized cells 40 25.5 0.13 Petrochemical 0.32 d

20 200 Immobilized cells 40 25.5 0.13 Carbohydrate 0.24 d

aAnnual ~.~t~ fOr ~~di”ary batch fermentation Were estimated from prOpr\@ary data. values obtained for ttre immobilized cell ertarnplea are COrnPutSd at 31.2 PerCent

below the comparable valuea for ordinary batch fermentation.

bAveraQe cost of petrochemical e~ual~ $0.17/lb. At ~ percent conversion efficiency, cost contribution of petrochemical eqUa[a $0.lQ/lb Of prOdUCt.

CAverWe coat of carbohydrate assumed at $o.04/lb of molaaees or $o.02/lb of cellulose-containing pellets from biomass reaidue. For ~ perCent free SUgar COntent Of

molasses, cost of sugar equals SO.06/lb. At 70 percent conversion efficiency from the sugar, cost contribution of molasses equals S0.1 I/lb of product. For 50 percent

cellulose content in the biomass pellets, coat of cellulose equala $0.04/lb. For 50 percent conversion efficiency to free sugar, followed by 70 percent conversion efficiency from the sugar, coat contribution of the pelleta also equals $0.1 lllb of product.

‘%hese unit costs maybe further reduced to S0.26 and $0.17/lb., respectively, for products whose annual U.S. production currently exceeds 1 billion lb. Asaumptlons include reduction in precursor cost by 20 percent (presumably because manufacturer controls SUPPlY of precursor); reduction in unit cost of immobilized cell process by

13 percent (d) and 42 percent (e), respectively; maximum of 60 percent product yield (e); and a nearly 100 percent bioconversion efficiency from the petrochemical

precursor.

SOURCE: Genex Corp.

Table l- B-2.-Basis for Estimating the Timetable for

Manufacture of Chemicals by Means of Microbial

Processes

And if bulk Assuming

If all the selling pricesc unit costsd

(in

Earliest date technologyb

(in 1979 1979 dollars)

for commercial is achieved dollars) equal equal or

production is: by: or exceed: exceed:

5 years 2 years $1.32/lb $0.66/lb

10 7 0.88 0.44

12 0.64 (0.43) 0.32(0.26)

20 17 0.48 (0.28) 0.24 (0.17)

qt IS ~SUrnad that development of the appropriate manufacturing facilltlm

begina at leaat 5 yeare prior to the onset of production.

%echnology refers to both genetic and biochemical engineering. Technology

would be achieved on demonstrating that the chemical could be biologically

produced In the laboratory at commercially desirable yields and reaction efficlenciea. Clt \S assurnad that all bulk selllng prices are rnSrkSd up 100 percent from the

corresponding unit costs, except for chemicals whose annual U.S. production

currently exceeds 1 billion lb. In those ca$es the bulk seillng prices (numbers

in parentheses) are assumed to be marked up only 67 pwcent.

dunit co8t8 were obtained from table l-B-l. See footnote of table I-B-1 for explanation of numbera in parentheses.

SOURCE: Genex Corp.

cient transformations of precursor to product, but

nothing exceptional with respect to current fermentation technology. Indeed, high product yields and

highly efficient reactions would be expected with

genetically engineered micro-organisms.

Two points should be stressed that place these

projections on the low side. First, they exclude certain groups of products, the end products of which

could not be microbially processed, although their

basic constituents could be produced microbiologically (e.g., monomers of microbial origin could form

chemically synthesized polymers). Second, the projections exclude naturally occurring products of

microbial origin, which could be effective or superior substitutes for chemically synthesized products

that could not he manufactured microbiologically. As

examples, dyes of microbial origin, such as prodigiosin, might advantageously replace those synthesized chemically, because their toxicity is lower than

their chemical counterparts. In the case of plastics, a

new generation of plastics of microbial origin, e.g.,

pullulans, would not have to be made from petrochemical feedstocks and would be biodegradable.

Explanation of tables

Tables l-B-3 through l-B-32 present the compounds

from two points of view. Tables l-B-3 to l-B-lo group

the compounds by industry subgrouped by product

category. Tables l-B-l 1 to I-B-32 group the cornpounds by product category irrespective of industry.

The tables based on industry present end use data

for each compound; e.g., in the pharmaceutical industry aspirin is listed as an aromatic used as an

analgesic, whereas in the chemical industry aniline is

listed as an aromatic used as a cyclic intermediate.

Thus, the similarities and differences between compounds of similar origin, i.e., product category, are

revealed. ●

The tables based solely on product category are

divided into two types; one type pertaining to market

data (tables 1-B-lo, 11, and the subsequent odd numbered ones through table l-B-33), and the other pertaining to technical data (the even numbered tables

from I-B-12 through l-B-32.)

The market data were obtained both from published sources and from prior proprietary studies
Appendix l-B—A Timetable for the Commercial Production of Compounds ● 277

Table l-B-3.—Pharmaceuticals: Small Molecules Table l=B=4.—Pharmaceuticais: Large Moiecuies

Product category End use Product category End use

Amino acids

Phenylalanine. . . . . . . . . . . .

Tryptophan. . . . . . . . . . . . . .

Arginine . . . . . . . . . . . . . . . .

Cysteine . . . . . . . . . . . . . . . .

Vitamins

Vitamin E. . . . . . . . . . . . . . . .

Vitamin B12 . . . . . . . . . . . . . .

Aromatics

Aspirin. . . . . . . . . . . . . . . . . .

p-acetaminophenol . . . . . . .

Steroid hormones

Corticoids

Cortisone . . .

Prednisone. .

Prednisolone

Aldosterone .

Androgens

Testosterone

Estrogens

Estradiol . . . .

Antibiotics

Penicillins. . . . .

Tetracycline . .

Cephalosporins

Short peptides

. . . . . . . . . .

. . . . . . . . . .

. . . . . . . . . .

. . . . . . . . . .

Intravenous solutions

Intravenous solutions

Therapeutic: liver disease

and hyperammonemia

Therapeutic: bronchitis and

nasal catarrh

Intravenous solutions,

prophylactic

Intravenous solutions

Analgesic

Analgesic

Therapeutic:

anti-inflammatory agent

Therapeutic:

ant i-inflammatory agent

Therapeutic:

anti-inflammatory agent

Therapeutic: control of

electrolyte imbalance

. . . . . . . . . . Therapeutic: infertility,

hypogonadism, and

hypopituitarism

. . . . . . . . . . Prophylactic, therapeutic:

vaginitis

. . . . . . . . . . Control of infectious diseases

. . . . . . . . . . Control of infectious diseases

. . . . . . . . . . Control of infectious diseases

Glycirie-Histidine-Lysine. . . Manufacturing processes:

tissue culture

SOURCE: Genex Corp.

perforlm?d t)y (k?llex. In the hitter case, data are L]st?d

that are not ~~roprietat”y ahhough the sources must

renuiin [;olli”i(l[?t~tiiil. Market values were estimated

})ov n~ultiplying the market ~olume (total amount of

product sold in 1978) by the bulk cost (unit bulk selling price in 1 980). Except for aromatics anci ali-

~)hatics, iill nmrket data represent worldwide estimates. hlarket data for aromatics and aliphatics are

ix?st rict[?d to the 1 Inited States. Data that could not be

found ~verc marked not available (N/A). Compouncls

\\’it h a high n]arket l’alue were identified, and those

[ha! could be pro(iuced biologically were selecte(i for

this report.

High market \alues \vcre relati~e to the incjustry

1111(1 end 11s(? list i tlg of each compound. For example,

\t’il h rcslxx’t to chemicals, normall.v only cyclic irl -

Peptide hormones

Insulin. . . . . . . . . . . . . . . . . . Control of diabetes

Endorphins. . . . . . . . . . . . . . Analgesics, narcotics,

prophylactics

Enkephalins . . . . . . . . . . . . . Analgesics, narcotics,

ACTHa 

. . . . . . . . . . . . . . . . . .

prophylactics

Diagnostic: adrenal instability

Glucagon . . . . . . . . . . . . . . . Therapeutic: diabetes-induced

hypoglycemia

Vasopressin . . . . . . . . . . . . . Therapeutic: antidiuretic

Human growth hormone. . . Therapeutic: dwarfism

Enzymes

Glucose oxidase . . . . . . . . . Diagnostic: measurement of

blood sugar

Urokinase . . . . . . . . . . . . . . . Therapeutic: antithrombotic

Asparaginase. . . . . . . . . . , . Therapeutic: antineoplastic

Tyrosine hydroxylase . . . . . Therapeutic: Parkinson’s

disease

Viral antigens

Hepatitis viruses . . . . . . . . . Vaccine

Influenza viruses . . . . . . . . . Vaccine

Herpes viruses. . . . . . . . . . . Vaccine

Varicella virus. . . . . . . . . . . . Vaccine

Rubella virus. . . . . . . . . . . . . Vaccine

Reoviruses . . . . . . . . . . . . . . Vaccine: common cold

Epstein-Barr virus . . . . . . . . Vaccine: infectious

mononucleosis,

nasopharyngeal

carcinoma, Burkitt

Iymphoma

Miscellaneous proteins

Interferon . . . . . . . . . . . . . . . Control of infectious diseases

Human serum albumin . . . . Therapeutic: shock and burns

Monoclinal antibodies . . . . Diagnostics: hepatitis, cancer,

etc; therapeutics

Gene preparations

Sickle-cell anemia . . . . . . . . Control of hereditary disorder

Hemophilias, . . . . . . . . . . . . Control of hereditary disorder

Thallasemias . . . . . . . . . . . . Control of hereditary disorder

%drenocofiicotroplc hormone.

SOURCE: Genex Corp.

termediat[?s with production volumes (which differ

from market volumes) exceeding 50 million lb were

selected, hut in the case of flavor iimi pertunw n]aterials, compounds with production values generally

exceeding 1 million lb were selected. In the case of

many pharmaceuticals, clinical importance was

weighed heavily in their selection process.

The technical data were also obtained both from

published and proprietary sources. \Vith respect to

the timetable for commercial production, the stated

Iengt h oft inw is the t i me required to de~elop exist ing

technolo~v (including both genetic and biochemicii]

engineering) to the point w’here it ma-v he applied to

ilppropriate man u fact uri ng faci Ii t ies f-or the klr~rescale pro(iuct ion of the desired compounds. These

time intervilis shoLl]d he sufficient for undertaking

76-565 0 - 81 - 19
278 ● Impacts of Applied Genetics —Micro-Organisms, Plants, and Animals

Table l- B-5.-Food Products Table l- B-6.-Agricultural Products

Product category End use

Amino acids

Glutamate . . . . . . . . . . . .

Cysteine . . . . . . . . . . . . .

Aspartame. . . . . . . . . . . . .

Vitamins

Vitamin C. . . . . . . . .

Vitamin D. . . . . . . . .

Aromatics

Benzoic acid . . . . . .

Aliphatics

Propionic acid. . . . .

Short peptides

Aspartame . . . . . . . .

Enzymes

Rennin

Amyloglucosidase.

a-amylase . . . . . . . .

Glucose isomerase.

Nucleotides

5

1

-IMPa. . . . . . . . . . .

5

1

-GMPb . . . . . . . . . .

,.. .

. . . .

. . . .

,.. .

,.. .

. . . .

. . . .

. . . .

. . . .

Food enrichment agent,

flavoring agent

Food enrichment agent,

manufacturing processes

Flavoring agent

Food additive, food

enrichment agent

Food enrichment agent

Food preservative

Food preservative

Artificial sweetener

Manufacturing processes

Manufacturing processes

Food enrichment agent,

manufacturing processes

Manufacturing processes:

sweetener

Flavoring agent

. . . . Flavoring agent

%’-lnosinic acid.

b51-guanylic acid.

SOURCE: Genex Corp.

all the R&D starting from the current knowledge

base necessary to demonstrate that the desired compounds can be biologically produced first in the

laboratory and then in the pilot plant at commercially desirable yields and reaction efficiencies. The

timetable does not consider delays caused by construction of new facilities nor delays required to

obtain Government regulatory approval of new

products.

It should be noted that in the technical data charts,

when glucose is listed as an alternate precursor by

fermentation, other carbohydrates, e.g., cellulose

and cornstarch, could be used. Moreover, if glucose

were the precursor of choice, the actual feedstock

would probably be a commodity like molasses as opposed to pure glucose.

Summary

Over 100 compounds representing 17 different

product categories that span the five industries

under evaluation are represented in table I-B-10. The

current market value of all these products exceeds

$27 billion. One particular compound, methane, accounts for over $12 billion. The even-numbered

Product category End use

Amino acids

Lysine. . . . . . . . . . . . . . . . . . . . .

Methionine. . . . . . . . . . . . . . . . .

Threonine. . . . . . . . . . . . . . . . . .

Tryptophan. . . . . . . . . . . . . . . . .

Vitamins

Nicotinic acid. . . . . . . . . . . . . . .

Riboflavin (B2

) . . . . . . . . . . . . . .

Vitamin C . . . . . . . . . . . . . . . . . .

Aliphatics

Sorbic acid. . . . . . . . . . . . . . . . .

Antibiotics

Penicillins . . . . . . . . . . . . . . . . .

Erythromycins. . . . . . . . . . . . . .

Peptide hormones

Bovine growth hormone. . . . . .

Porcine growth hormone . . . . .

Ovine growth hormone. . . . . . .

Viral antigens

Foot-and-mouth disease virus.

Feed additive

Feed additive

Feed additive

Feed additive

Feed additive

Feed additive

Feed additive

Feed preservative

Feed additive, prophylactic

Feed additive, prophylactic

Growth promoter

Growth promoter

Growth promoter

Vaccine

Rous sarcoma virus . . . . . . . . . . Vaccine

Avian Ieukemiavirus . . . . . . . . . Vaccine

Avian myeloblastosis virus. . . . Vaccine

Enzymes

Papain . . . . . . . . . . . . . . . . . . . . . Feed additive

Glucose oxidase . . . . . . . . . . . . Feed preservative

Pesticides

Microbial . . . . . . . . . . . . . . . . . . Insecticide

Aromatic. . . . . . . . . . . . . . . . . . . Insecticide

Inorganic

Ammonia . . . . . . . . . . . . . . . . . . Fertilizer

SOURCE: Genex Corp.

tables from 1-B-12 to I-B-32 project that within 20

years all these products could be manufactured

using genetically engineered microbial strains on a

more economical basis than using today’s conventional technologies. In many cases, the time required

to apply genetically engineered strains in commercial

fermentations could be reduced to as little as 5 years.

The impact of genetic engineering on selected

markets is shown in table I-B-33. only five product

categories are considered here, and in one, amino

acids, only a few of the compounds comprising it are

evaluated. The products represented in the five categories currently have a total market value exceeding

$800 million. However, within 20 years this market

value could rise to over $5 billion (in 1980 dollars)

due largely to the application of genetic engineering.

In a number of cases, the desired products would

most likely not be available in significant quantities if

not for the application of genetic engineering technology.
Appendix l-B—A Timetable for the Commercial Production of Compounds ● 279

Table l- B-7.—Chemicals: Aliphatics

Compound End use

Acetic acida . . . . . . . . . . . . . . . .

Acrylic acida

. . . . . . . . . . . . . . . .

Adipic acida 

. . . . . . . . . . . . . . . .

Bis (2-ethylhexyl) adipate. . . . .

Citronella. . . . . . . . . . . . . . . . . .

Citronellol . . . . . . . . . . . . . . . . .

Ethanol a

. . . . . . . . . . . . . . . . . . .

Ethanolamine. . . . . . . . . . . . . . .

Ethylene glycola 

. . . . . . . . . . . .

Ethylene oxidea

. . . . . . . . . . . . . .

Geraniol . . . . . . . . . . . . . . . . . . .

Glycerol a 

. . . . . . . . . . . . . . . . . .

Isobutylene . . . . . . . . . . . . . . . .

Itaconic acid . . . . . . . . . . . . . . .

Linalool. . . . . . . . . . . . . . . . . . . .

Linalyl acetate. . . . . . . . . . . . . .

Methane. . . . . . . . . . . . . . . . . . .

Nerol. . . . . . . . . . . . . . . . . . . . . .

Pentaerythritol. . . . . . . . . . . . . .

Propylene giycola 

. . . . . . . . . . .

Sorbitol. . . . . . . . . . . . . . . . . . . .

a-terpineol . . . . . . . . . . . . . . . . .

a-terpinylacetate . . . . . . . . . . .

Miscellaneous acyclic

Miscellaneous acyclic

Misceilaneous acyclic

Plasticizer

Flavor/perfume material

Flavor/perfume material

Miscellaneous acylic

Miscellaneous acyclic

Miscellaneous acyclic

Miscellaneous acyclic

Flavor/perfume material

Miscellaneous acyclic

Miscellaneous acyciic,

flavor/perfume material

Plastics/resin

Flavor/perfume material

Flavor/perfume material

Primary petroleum product

Flavor/perfume material

Miscellaneous acyclic

Miscellaneous acyiic

Miscellaneous acyclic

Flavor/perfume material

Flavor/perfume material

alndicates compounds also identified by the Massachusetts Institute of

Technology. The following additional chemicals were identified by MIT as

amenable to biotechnological production methods: acetaldehyde, acetoin,

acetone, acetylene, acrylic acid, butadiene, butano~ butyl acetate,

butyraldehyde, dihydroxyacetone, ethyl acetate, ethyl acrylate, ethylene, formaldehyde, isoprene, isopropanol, methanol, methyl ethyl ketone, methyl

acrylate, propylene, propylene oxide, styrene, vinyl acetate.

SOURCE: Genex Corp. and the Massachusetts Institute of Technology.

Table l-B-8.—Chemicals: Aromatics and

Miscellaneous

Product category End use

Aromatics

Aniline . . . . . . . . . . . . . . . . . . . . Cyclic intermediate

Benzoic acid. . . . . . . . . . . . . . . . Cyclic intermediate

Cresols. . . . . . . . . . . . . . . . . . . . Cyclic intermediate

Phenol . . . . . . . . . . . . . . . . . . . . Cyclic intermediate

Phthalic anhydride . . . . . . . . . . Cyclic intermediate

Cinnamaldehyde. . . . . . . . . . . . Flavor/perfume material

Diisodecyl phthalate. . . . . . . . . Plasticizer

Dioctyle phthalate. . . . . . . . . . . Plasticizer

Inorganic

Ammonia . . . . . . . . . . . . . . . . . . Manufacturing processes

Hydrogen . . . . . . . . . . . . . . . . . . Manufacturing processes

Enzymes

Pepsin. . . . . . . . . . . . . . . . . . . . . Manufacturing processes

Bacillus protease. . . . . . . . . . . . Manufacturing processes

Mineral leaching

Transition metals (cobalt, Inorganic intermediates;

nickel, manganese, iron). . . . . . catalysts

Biodegradation . . . . . . . . . . . . . Removal of organic

phosphates, aryl

sulfonates, and

haloaromatics

Table l- B-9.—Energy Products

Product category End use

Enzymes

Ethanol dehydrogenase. Manufacturing processes

Hydrogenate. . . . . . . . . . Manufacturing processes

Biodegradation. . . . . . . . Petroleum byproducts removal

Aliphatics

Methane. . . . . . . . . . . . . . Fuel

Ethanol . . . . . . . . . . . . . . Fuel

Inorganic

Hydrogen. . . . . . . . . . . . . Fuel

Mineral leaching

Uranium. . . . . . . . . . . . . . Fuel

SOURCE: Genx Corp.

Table I- B- 10.—Total Market Values for the

Various Product Categories

Number of Current value

Product category compounds ($ millions)

Amino acids. . . . . . . . . . . . . . . . 9 $ 1,703.0

Vitamins. . . . . . . . . . . . . . . . . . . 6 667.7

Enzymes. . . . . . . . . . . . . . . . . . . 11 217.7

Steroid hormones . . . . . . . . . . . 6 376.8

Peptide hormones. . . . . . . . . . . 9 263.7

Viral antigens. . . . . . . . . . . . . . . 9 N/A

Short peptides. . . . . . . . . . . . . . 2 4.4

Nucleotides. . . . . . . . . . . . . . . . 2 72.0

Miscellaneous proteins . . . . . . 2a 300.0

Antibiotics. . . . . . . . . . . . . . . . . 4

b

4,240.0

Gene preparations. . . . . . . . . . . 3 N/A

Pesticides . . . . . . . . . . . . . . . . . 2

b

100.0

Aliphatics:

Methane. . . . . . . . . . . . . . . . . 12,572.0

Other. . . . . . . . . . . . . . . . . . . . 24c

2,737.5

Aromatics. . . . . . . . . . . . . . . . . . 10c

1,250.9

Inorganic . . . . . . . . . . . . . . . . . 2 2,681.0

Mineral leaching . . . . . . . . . . . . 5 N/A

Biodegradation . . . . . . . . . . . . . N/A N/A

Totals . . . . . . . . . . . . . . . . . 107 $27,186.7d

%nly two of a number of compounds are considered here.

%hese numbers refer to major classes of compounds; not actual numbers of

compounds.

cThege numbers refer only to those compounds rePresenting the k9est

market volume In classes specified in the text.

dcument value excluding methane = $14,614,7W,~.

SOURCE: Genex Corp.

SOURCE: Genex Corp.
280 ● Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

Table I-B-11.-Amino Acids: Market Information

Current market data

Market

volume Bulk cost Market value

Compound 1,000 lb $/lb ($ millions)

Arginine . . . . . . . . 900 12.73 11.46

Aspartate . . . . . . . 3,000 2.86 8.6

Cysteine. . . . . . . . 600 22.75 13.6

Glutamate. . . . . . . 600,000 1.80 1,080.0

Lysine . . . . . . . . . 129,000 2.10 258.0

Methionine. . . . . . 210,000 1.40 294.0

Phertylalanine. . . . 300 38.18 11.46

Threonine. . . . . . . 300 58.18 16.2

Tryptophan. . . . . . 225 43.18 9.71

SOURCE: Compiled by Genex Corp. from data in references 1,2, and 3.

Table l-B-12.—Amino Acids: Technical Information

Is precursor Time to implement

renewable/non- Alternate commercial fermentation by

Typical synthetic renewable precursor by genetically engineered

Compound process Typical precursor limited fermentation strain

Arginine . . . . . . . .

Aspartate . . . . . . .

Cysteine . . . . . . . .

Glutamate. . . . . . .

Lysine. . . . . . . . . .

Methionine. . . . . .

Phenylalanine. . . .

Threonine. . . . . . .

Tryptophan. . . . . .

fermentation

fermentation

extraction

fermentation

fermentation

chemical

chemical

fermentation

chemical

glucose

and NH4

+a

fumaric acid

and ammonia

protein

hydrolysis

glucose

and NH4

+

glucose

and NH4

+

B-methylmercapto

proplonaldehyde

a-acetaminocinnamic acid

glucose

and NH4

+

a-ketoglutaric

phenylhydrazone

renewable

limited

renewable

renewable

renewable

nonrenewable

limited

renewable

nonrenewable

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

5 yrs.

5 yrs.

5 yrs.

5 yrs.

5 yrs.

10 yrs.

5 yrs.

5 yrs.

5 yrs.

5 yrs.

aArnmonlum ion.

SOURCE: Compiled by Genex Corp. from data in references 2,3,4, and 5

Table l- B-13.-Vitamins: Market Information

Current market data

Market

volume Bulk cost Market value

Compound 1,000 lb $/lb ($ millions)

Nicotinic acid. . . . 1,400 1.82 2.5

Riboflavin (B2). . . . 22 15.40 0.34

Vitamin B12 . . . . . . 6,991.60 153.8

Vitamin C . . . . . . . 90,0% 4.50 405.0

Vitamin D . . . . . . . 12 42.50 0.51

Vitamin E . . . . . . . 3,641 29.00 105.6

SOURCE: Compiled by Genex Corp. from data in references 1,6,7,8, and 9.
Appendix l-B—A Timetable for the Commercial production of Compounds . 281

Table l- B-14.—Vitamins: Technical Information

Is precursor Time to implement

renewable/non- Alternate commercial fermentation by

Typical synthetic renewable precursor by genetically engineered

Compound process Typical precursor limited fermentation strain

Nicotinic Acid . . . chemical aikyl a-subst. nonrenewable glucose 10 yrs.

and NH4

+a

Riboflavin (B2). . . . fermentation pyridines renewable — 10 yrs.

glucose

Vitamin B12 . . . . . . fermentation carbohydrates renewable — 10 yrs.

Vitamin C . . . . . . . semisynthetic glucose or renewable 10 yrs.

sorbitol

Vitamin D . . . . . . . fermentation glucose renewable glucose 10 yrs.

Vitamin E . . . . . . . extraction wheat germ oil limited glucose 15 yrs.

aAmmonium ion,

SOURCE: Compiled by Genex Corp. from data in references 4,7,8, 10,11, and 12.

Table I-B-1 5.—Enzymes: Market Information

Current market data

Market

volume Bulk cost Market value

Compound 1,000 lb $/lb ($ millions)

a-amylase. , . . . . . 600 19.33 11.6

Amyloglucosidase 600

Asparaginase. . . . (lnformation not available)

Bacillus protease. 1,000 8.28 8.2

Ethanol

dehydrogenase . (Information not available)

Glucose isomerase 100 40000 400

Glucose oxidase . 0.80

Hydrogenate . . . . (Iformation not available)

Papain. . . . . . . . . . 200 59.00 11.8

Pepsin. . . . . . . . . . 10 360.00 3.8

Rennin. . . . . . . . . . 24 696.00 40.0

Tyrosine

hydroxylase . . . . (Information not available)

Urokinase. . . . . . . 60,000 IUa

89.5

alu = international units.

SOURCE: Compiled by Genex Corp. from data in references 9,13,14,15, and 16.
282 ● Impacts of Applied Genetics—Micro-Organ/sins, Plants, and Animals

Table I- B-16.-Enzymes: Technical Information

Is precursor Time to implement

renewable/non- Alternate commercial fermentation by

Typical synthetic renewable precursor by genetically engineered

Compound process Typical precursor limited fermentation strain

a-amylase. . . . . . .

Amyloglucosidase

Asparaginase. . . .

Bacillus protease.

Ethanol

dehydrogenase

Glucose

isomerase. . . . .

Glucose oxidase .

Hydrogenate . . . .

Papain. . . . . . . . . .

Pepsin. . . . . . . . . .

Rennin. . . . . . . . . .

Tyrosine . . . . . . . .

Urokinase. . . . . . .

fermentation

fermentation

extraction

fermentation

fermentation

fermentation

extraction

fermentation

fermentation

extraction

extraction

molasses

molasses

tissue culture

molasses

(Information not available)

glucose

and NH4

+a

molasses

(Information not available)

papaya

molasses

molasses

tissue culture

tissue culture

renewable

renewable

renewable

renewable

renewable

renewable

renewable

renewable

renewable

renewable

renewable

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

5 yrs.

5 yrs.

5 yrs.

5 yrs.

10 yrs.

5 yrs.

5 yrs.

10 yrs.

5 yrs.

5 yrs.

5 yrs.

5 yrs.

5 yrs.

aArnrnonium ion.

SOURCE: Compiled by Genex Corp. from data in references 4,5,13,14,16,17, and 18.

Table l=B=17.-Sterold Hormones: Market

information

Current market data

Market

volume Bulk cost Market value

Compound 1,000 lb $/lb ($ millions)

Corticoids. . . . ● . . 305.8

Cortisone . . . . . . . N/A 208.84 N/A

Prednisone. . . . . . N/A 467.62 N/A

Prenisolone . . . . . N/A 463.06 N/A

Aldosterone . . . . . N/A N/A N/A

Androgens 10,8

Testosterone . . . . (Information not available)

Estrogens. . . . . . . 60.2

Estradiol . . . . . . . . (Information not available)

SOURCE: Compiled by Genex Corp. from data In references 1 and 4.
Appendix l-B—A Time table for the Commercial Production of Compounds ● 283

Table I-B-18.—Steroid Hormones: Technical Information

Is precursor Time to implement

renewable/non- Alternate commercial fermentation by

Typical synthetic renewable precursor by genetically engineered

Compound process Typical precursor limited fermentation strain

Corticoids

Cortisone

Prednisone. . . . . .

Predisolone

Aldosterone

Androgens

Testosterone . . . .

Estrogens

Estradiol . . . . . . . .

semisynthetic diosgenin or renewable glucose 10 yrs.

stigmasterol

semisynthetic chemical renewable glucose 10 yrs.

modification

of cholesterol

semisynthetic chemical renewable glucose 10 yrs.

modification of

cholesterol

SOURCE: Compiled by Genex Corp. from data in references 4,19,20,21, and 22.

.

Table l- B-19.—Peptide Hormones: Market

Information

Current market data

Compound

ACTHa . . . . . . . . . .

Bovine growth

hormone. . . . . .

Endorphins. . . . . .

Enkephalins . . . . .

Glucagon . . . . . . .

Human growth

hormone. . . . . .

Insulin. . . . . . . . . .

Ovine growth

hormone. . . . . .

Porcine growth

hormone. . . . . .

Vasopressin. . . . .

Market

volume Bulk cost Market value

1,000 lb $/lb ($ millions)

N/A N/A 5.6

0.0 0.0

(lnformation not available)

(Information not available)

(Information not available)

N/A N/A 75.0

N/A N/A 163.1

0.0 0.0 0.0

0.0 0.0

(lnformation not available)

aAdrenoco~icotropic hormone.

SOURCE: Compiled by Genex Corp. from data in reference 4.
284 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Table I- B-20.-Peptide Hormones: Technical Information

Is precursor Time to implement

renewable/non- Alternate commercial fermentation by

Typical synthetic renewable precursor by genetically engineered

Compound process Typical precursor limited fermentation strain

ACTHa. . . . . . . . . .

Bovine growth

hormone. . . . . .

Endorphins. . . . . .

Enkephalins. . . . .

Glucagon . . . . . . .

Human growth

hormone. . . . . .

Insulin. . . . . . . . . .

Ovine growth

hormone. . . . . .

Porcine growth

hormone. . . . . .

Vasopressin. . . . .

extraction

extraction

extraction

extraction

extraction

extraction

extraction

extraction

extraction

extraction

adrenal cortex

anterior pituitary

brain

brain

pancreas

anterior pituitary

pancreas

anterior pituitary

anterior pituitary

posterior pituitary

limited

limited

limited

limited

limited

limited

limited

limited

limited

limited

glucose

and NH4

+b

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose,

5 yrs.

5 yrs.

5 yrs.

5 yrs.

5 yrs.

5 yrs.

5 yrs.

10 yrs.

10 yrs.

5 yrs.

and NH4

+

aArjren~@iCOt@C hormone.

bAmmonium ion.

SOURCE: Compiled by Genex Corp. from data in references 4,23, and 24.

Table l- B-21.—Vjral Antigens: Market Information

Current market data

Market

volume Bulk cost Market value

Compound 1,000 lb $/lb ($ millions)

Avian leukemia

virus. . . . . . . . . . . (Information not available)

Avian myeloblastosis

virus. . . . . . . . . . . (information not available)

Epstein-Barr virus 0.0 0.0 0.0

Hepatitis virus . . . 0.0 0.0 0.0

Herpes virus. . . . . 0.0 0.0 0.0

Hoof and mouth

disease virus . . 0.0

influenza virus . . . (lnformation not available)

Reoviruses . . . . . . 0.0 0.0 0.0

Rous sarcoma

virus. . . . . . . . . . . (Information not available)

Rubella virus. . . . . (Information not available)

Varicella virus . . . (information not available)

SOURCE: Compiled by Genex Corp. from data in reference 4.
Appendix I-B—A Timetable for the Commercial Production of Compounds ● 2 8 5

Table l- B-22.—Viral Antigens: Technical Information

Is precursor Time to implement

renewable/non- Alternate commercial fermentation by

Typical synthetic renewable precursor by genetically engineered

Compound process Typical precursor limited fermentation strain

Avian leukemia. . .

virus

Avian

myeloblastosis.

virus

Epstein-Barr virus

Hepatitis. . . . . . . .

viruses

Herpes . . . . . . . . .

viruses

Hoof and mouth. .

disease virus

Influenza. . . . . . . .

viruses

Reoviruses . . . . . .

Rous sarcoma . . .

virus

Rubella . . . . . . . . .

virus

Varicella . . . . . . . .

virus

(Information not available)

(Information not available)

tissue culture Iymphoblasts

(Information not available)

(Information not available)

(Information not available)

(Information not available)

(Information not available)

(Information not available)

tissue culture duck embryonic

cells

(Information not available)

glucose

and NH4

+a

glucose

and NH4

+

renewable glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

glucose

and NH4

+

renewable glucose

and NH4

+

glucose

NH4

+

5 yrs.

5 yrs.

5 yrs.

5 yrs.

5 yrs.

5 yrs.

10 yrs.

15 yrs.

5 yrs.

5 yrs.

5 yrs.

aAmmonium ion.

SOURCE: Compiled by Genex Corp. from data in references 4 and 25.

Table l-B-23.—Short Peptides, Nucleotides, and

Miscellaneous Proteins: Market Information

Current market data

Market

volume Bulk cost Market value

Product category 1,000 lb $/lb ($ millions)

Short peptidesa

Aspartame . . . . . . 40 110.00 4.4

Glycine-histidineIysine. . . . . . . . . . (Information not available)

Nucleotides b

5

1

-IMP c . . . . . . . . . 4,000 12.00 48.0

5

1

-GMP d . . . . . . . . 2,000 12.00 24.0

Miscellaneous proteinsa

Interferon . . . . . . . N/A N/A 50.0

Human serum

albumin. . . . . . . . 250 1,000.00 250.0

Monoclinal

antibodies. . . . . . (Information not available)

aData from references 4 and 26.

bData from references 4 and 27.

c5’-inosinic acid.

d5’-guanylic acid.

‘Data from reference 4.

SOURCE: Compiled by Genex Corp.
286 ● Impacts of Applied Genet/cs—Micro-Organisms, Plants, and Animals

Table l- B-24.—Short Peptides, Nucleotides, and Miscellaneous Proteins: Technical information

Is precursor Time to implement

renewable/non- Alternate commercial fermentation by

Typical synthetic renewable precursor by genetically engineered

Product category process Typical precursor limited fermentation strain

Short peptidesa

Aspartame . . . . . . chemical phenylalanine & renewable glucose 5 yrs.

aspartic acid and NH4

+

Glycine-histidineIysine. . . . . . . . . . . extraction human serum renewable glucose 5 yrs.

and NH4

+

Nucleotides c

5

1

-lMP d . . . . . . . . . extraction yeast renewable glucose 10 yrs.

and NH4

+

5

1

-GMPe. . . . . . . . . extraction yeast renewable glucose 10 yrs.

and NH4

+

Mlscellaneous proteinsf

Interferon . . . . . . . extraction or leukocytes, renewable glucose 5 yrs.

tissue culture Iymphoblasts, and NH4

+

or fibroblasts

Human serum

albumin . . . . . . . . . extraction human serum renewable glucose 5 yrs.

and NH4

+

Monoclinal

antibodies . . . . . somatic cell various cells renewable glucose 10 yrs.

hybridization and NH4

+

%ata from reference 4 and 27.

bAmmonium ion.

cData from references 4 and ~.

%’-inoalnic acid.

‘5’-guanyiic acid.

‘Data from reference 4.

SOURCE: Compiied by Genex Corp.

Table I- B-25.-Antibiotics, Gene Preparations, and

Pesticides: Market information

Current market data

Market

volume Bulk cost Market value

Product category 1,000 lb $/Ib ($ millions)

Antibiotic@

Penicillins. . . . . . . 49,300 22.11 1,080.0

Tetracycline. . . . 29,300 34.13 1,000.O

Cephalosporins. . 4,210 114.00 460.0

Erythromycins . . . (Information not available)

Gene preperationsb

Sickle cell anemia 0.0 0.0 0.0

Hemophilias. . . . . 0.0

Thallasemias . . . . 0.0 0.0 0.0

Pesticidesc

Microbial. . . . . . . . N/A N/A 25.0

Aromatics . . . . . . . N/A N/A 75.0

aData from references 4,28, and 9.

bData from references 4 and X.

cData from references 4 and ~.

SOURCE: Compiled by Genex Corp.
Appendix I-B—A Timetable for the Commercial Production of Compounds ● 287

Table l- B-26.—Antibiotics, Gene Preparations, and Pesticides: Technical Information

Is precursor Time to implement

renewable/non- Alternate commercial fermentation by

Typical synthetic renewable precursor by genetically engineered

Product category process Typical precursor limited fermentation strain

Antibiotics

Penicillins. . . . .

Tetracycline . .

Cephalosporins

Erythromycins .

. .

. .

. .

Gene preprationsb

Sickle cell anemia

Hemophilias. . . . .

Thallasemias . . . .

Pestlcidesc

Microbial. . . . . . . .

Aromatics. . . . . . .

fermentation lactose &

semisynthetic nitrogenous oils

fermentation lactose &

nitrogenous oils

fermentation lactose &

nitrogenous oils

fermentation lactose &

nitrogenous oils

(No process exists

currently)

(No process exists

currently)

(No process exists

currently)

fermentation molasses &

fishmeal

semi synthetic naphthalene

limited

limited —

limited

limited

glucose

and NH4

+d

glucose

and NH4

+

glucose

and NH4

+

renewable

nonrenewable —

10 yrs.

10 yrs.

10 yrs.

10 yrs.

15 yrs.

20 yrs.

20 yrs.

5 yrs.

10 yrs.

aData from references 4,5,28, 3~t and 32. Coata from references 4 and ~.

bData from reference 4. ‘Ammonium ion.

SOURCE: Compiled by Genex Corp.

Table l= B-27. -AIiphatics: Market Information

Current market data

Market

volume Bulk cost Market value

Compound 1,000 lb $/lb ($ millions)

Acetic acid . . . . . . 823,274

Acrylic acid. . . . . . 46,503

Adipic acid . . . . . . 181,097

Bis (2-ethylehexyl) 43,015

adipate

Citronella . . . . . . . 394

Citronellol. . . . . . . 1,443

Ethanol . . . . . . . . . 1,048,000

Ethanolamine. . . . 320,236

Ethylene glycol . . 3,137,000

Ethylene oxide . . . 525,113

Geraniol . . . . . . . . 2,307

Glycerol . . . . . . . . 116,612

Isobutylene. . . . . . 597,712

Itaconic acid. . . . . 200

Linalool. . . . . . . . . 3,341

Linalyl acetate . . . 1,535

Methane . . . . . . . . 878,000,000

Nerol. . . . . . . . . . . 462

Pentaerythritol. . . 117,085

Propionic acid . . . 62,848

Propylene glycol . 525,527

Sorbic acid . . . . . . 20,000

Sorbitol. . . . . . . . . 160,267

a-terpineol . . . . . . 2,416

a-terpinyl acetate. 1,066

0.23

0.43

0.50

0.49

3.90

4.50

0.24

0.46

0.31

3.25

0.54

0.95

0.83

2.60

3.50

0.013

4.20

0.62

0.21

0.73

2.15

0.36

1.28

1.30

189.4

20.0

90.5

21.1

6.5

251.5

147.3

972.5

189.0

7.5

63.0

567.2

0.2

8.7

5.4

11,573.0

1.9

72.6

13.2

173.4

43.0

57.7

3.0

1.4

SOURCE: Compiled by Genex Corp. from data in references 1,4,9, and 33.
288 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Table l- B-28.—Aliphatics: Technical Information

Is precursor Time to implement

renewable/non- Alternate commercial fermentation by

Typical synthetic renewable precursor by genetically engineered

Compound process Typical precursor limited fermentation strainb

Acetic acid. . . . . . . . .

Acrylic acid . . . . . . . .

Adipic acid. . . . . . . . .

Bis (2-ethylhexyl). . . .

adipate

Citronella . . . . . . . . .

Citronellol . . . . . . . . .

Ethanol. . . . . . . . . . . .

Ethanolamine . . . . . .

Ethylene glycol . . . . .

Ethylene oxide. . . . . .

Geraniol . . . . . . . . . . .

Glycerol . . . . . . . . . . .

Isobutylene . . . . . . . .

Itaconic acid . . . . . . .

Linalool . . . . . . . . . . .

Linalyl acetate . . . . . .

Methane. . . . . . . . . . .

Nerol. . . . . . . . . . . . . .

Pentaerythritol . . . . .

Propionic acid . . . . . .

Propylene glycol . . . .

Sorbic acid. . . . . . . . .

Sorbitol. . . . . . . . . . . .

a-terpineol . . . . . . . . .

a-terpinyl acetate . . .

chemical

chemical

chemical

chemical

chemical

chemical

chemical

chemical

chemical

chemical

chemical

chemical

chemical

fermentation

chemical

chemical

chemical

chemical

chemical

chemical

chemical

chemical

chemical

chemical

chemical

methanol

or ethanol

ethylene

phenol

phenol

isobutylene

isobutylene

ethylene

ethylene

ethylene

ethylene

isobutylene

soap manuf.

petroleum

molasses

isobutylene

isobutylene

natural gas

isobutylene

acetaldehyde &

formaldehyde

ethanol &

carbon monoxide

propylene

crotonaldehyde &

malonic acid

glucose

isobutylene

isobutylene

nonrenewable

nonrenewable

nonrenewable

nonrenewable

nonrenewable

nonrenewable

nonrenewable

nonrenewable

nonrenewable

nonrenewable

nonrenewable

nonrenewable

nonrenewable

renewable

nonrenewable

nonrenewable

nonrenewable

nonrenewable

nonrenewable

limited

nonrenewable

nonrenewable

renewable

nonrenewable

nonrenewable

glucose

glucose

glucose

glucose

glucose

glucose

glucose

glucose

glucose

glucose

glucose

glucose

glucose

. . . 

glucose

glucose

sewage

glucose

glucose

glucose

glucose

glucose

glucose

glucose

10 yrs.

10 yrs.

10 yrs.

20 yrs.

20 yrs.

20 yrs.

5 yrs.

10 yrs.

5 yrs.

5 yrs.

20 yrs.

5 yrs.

10 yrs.

5 yrs.

20 yrs.

20 yrs.

10 yrs.

20 yrs.

10 yrs.

10 yrs.

10 yrs.

15 yrs.

10 yrs.

20 yrs.

20 yrs.

%Vherever glucose ia mentioned, other carbohydrates maybe aubatituted, including starch and cellulose.

bln many Casea these times are bgsed on more readily developed fermentations using nonrenewable or limited hydrocarbons as Precursors.

SOURCE: Compiied by Genex Corp. from data in references 4,33,34, and 35.

Table I= B-29.-Aromatics: Market Information

Current market data

Market

volume Bulk cost Market value

Compound 1,000 lb $/lb ($ millions)

Aniline . . . . . . . . . . . . 187,767 0.42 78.9

Aspirin . . . . . . . . . . . . 32,247 1.41 45.5

Benzoic acid . . . . . . . 36,822 0.47 17.3

Cinnamaldehyde. . . . 1,098 2.10 3.4

Cresols. . . . . . . . . . . . 94,932 0.54 51.2

Diisodecyl

phthalate . . . . . . . . 151,319 0.42 63.6

Dioctyl phthalate. . . . 391,131 0.42 164.3

p-acetaminophenol. . 20,000 2.65 53.0

Phenol . . . . . . . . . . . . 1,431,000 0.36 515.2

Phthalic anhydride . . 646,289 0.40 258.5

SOURCE: Compiled by Genex Corp. from data in references 1 and 9.
Appendix l-B—A Timetable for the Commercial Production of Compounds ● 289

Table l- B-30.-Aromatics: Technical Information

Is precursor

renewable/nonTypical synthetic renewable

Compound process Typical precursor limited

Aniline . . . . . . . . . . . . chemical benzene nonrenewable

Aspirin . . . . . . . . . . . . chemical phenol nonrenewable

Benzoic acid . . . . . . . chemical tar oil nonrenewable

Cinnamaidehyde. . . . chemical benzaldehyde nonrenewable

acetaldehyde

Cresols. . . . . . . . . . . . chemical phthalic nonrenewable

an hydride

Diisodecyl . . . . . . . . . chemical coal tar nonrenewable

phthalate

Dioctyl . . . . . . . . . . . . chemical coal tar nonrenewable

phthalate

p-acetaminophenol. . chemical nitrobenzene nonrenewable

Phenol . . . . . . . . . . . . chemical coal tar nonrenewable

Phthalic . . . . . . . . . . . chemical coal tar nonrenewable

anhydride

Time to implement

Alternate commercial fermentation by

precursor by genetically engineered

fermentation strain

aromatica10 yrs.

aromatic 5 yrs.

aromatic 10 yrs.

aromatic 20 yrs.

aromatic 10 yrs.

aromatic 20 yrs.

aromatic 20 yrs.

aromatic 5 yrs.

aromatic 10 yrs.

aromatic 15 yrs.

aAromatic refers t. benzene or benzene derivative. Eventually it is anticipated that Iignin, a renewable resource, would sewe as a Precursor.

SOURCE: Compiled by Genex Corp. from data in references 4 and 35.

Table I-B-31 .—lnorganics and Mineral Leaching:

Market Information

Current market data

Market

volume Bulk cost Market value

Product category 1,000 lb $/lb ($ millions)

Inorganic

Ammonia . . . . . . . 33,400,000 0.06 2,004.0

Hydrogen . . . . . . . 451,000 0.15 677.0

Mineral leaching

Uranium . . . . . . . . (Information not available)

Transition metals. (Information not avaiiable)

(cobalt, nickel, manganese, iron)

SOURCE: Compiled by Genex Corp. from data in reference 4.

Table l- B-32.-lnorganics and Mineral Leaching: Technical Information

Is precursor Time to implement

renewable/non- Alternate commercial fermentation by

Typical synthetic renewable precursor by genetically engineered

Product category process Typical precursor limited fermentation strain

Inorganic

Ammonia . . . . . . . chemical water and coke nonrenewable nitrogen(air) 15 yrs.

Hydrogen . . . . . . . catalytic petroleum nonrenewable water and air 15 yrs.

reforming

Mineral leaching

Uranium . . . . . . . . (Information not available)

Transition metals. (Information not available)

(cobalt, nickel, manganese, iron)

SOURCE: Compiled by Genex Corp. from data in references 4 and 35.
290 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Table l- B-33.-Projected Growth of Selected Markets

Involving Applications of Genetic Engineering

Projected

market

Current market in 20 yrs.

Product category $ millions $ millions

Amino acidsa

. . . . . . . . . . 300

Miscellaneous proteins. 300 1,000

Gene preparations . . . . . 0

Short peptides . . . . . . . . 5 2,100

Peptide hormones . . . . .

Totals . . . . . . . . . . . . . . 865 5,100

%nly four amino acids are considered here.

SOURCE: Genex Corp.

References

1.

2.

3.

4.

5.

6.

7.

8.

9.

10.

11.

Chemical Marketing Reporter, March and April

1980.

Hirose, Y., and Okada, H., “Microbial Production

of Amino Acids, ” in Microbial Technology: Microbial Processes, vol. 1, H. J. Peppier and D. Perlman

(eds.) (New York: Academic Press, 1979), pp.

211-240.

Hirose, Y., Sane, K., and Shibai, H., “Amino

Acids, ” in Annual Reports on Fermentation Processes, vol. 2, (;. T. Tsao and D. Perlman (eds.)

(New York: Academic Press, 1978), pp. 155-190.

(;enex Corp. proprietary information.

Weinstein, L., “Chemotherap~ of Microbial Diseases, ” in The Pharmacological Basis of Therapeutics, 5th cd., L. S. Goodman and A. Gilman (eds.)

(New York: MacMillan Publishing Co., Inc., 1975),

pp. 1,090-1,247.

Cohn, V. H., “k’at-Soluble Vitamins: Vitamin K and

vitanlin E,” in The Pharmacological Basis of Therapeutics, 5th cd., L. S. Gmodrnan and A. Gilman

(eds.) (New York: MacMillan Publishing Co., Inc.,

1975), PP. 1,591-1,600.

Florent, J., and Nitiet, L., “Vitamin B,z,” in A4icrobial Technology Microbial Processes, vol. 1, H. J.

Peppier and D. Perlman (eds.) (New York: Academic Press, 1979), pp. 497-520.

Perlman, D., “Microbial Process for Riboflavin

Product ion,” in Microbial Technology: Microbial

Processes, vol. 1, H. J. Peppier (eds.) (New York:

Academic Press, 1979), pp. 521-528.

Synthelic Organic Chemicals, March and April

1980.

Burns, J. J., “Water Soluble Vitamins, The Vitamin B Complex, “ in The Pharmacological Basis of

Therapeutics, 5th cd., L. S. Goodman and A.

Gilrnan (eds.) (New York: MacMillan Publishing

Co., Inc., 1975), pp. 1,549-1,563.

Greengard, P., “Water Soluble Vitamins, The Vita12

13

min B Complex, “ in The Pharmacological Basis of

Therapeutics, 5th cd., L. S, Goodman and A.

Gilman (eds.) (New York: MacMillan Publishing

Co., Inc., 1975), pp. 1,549-1,563.

Straw, J. A., “Fat-Soluble Vitamins: Vitamin D,” in

The Pharmacological Basis of Therapeutics, 5th

cd., L. S. Goodman and A. Gilman (eds.) [New

York: MacMillan Publishing Co., Inc., 1975), pp.

1,579-1,590.

Aunstrup, K., “Industrial Approach to Enzyme

Product ion,” in Biotechnological Applications of

Pro(eins and Enzymes, Z. Bohak and N. Sharon

(eds.) (New York: Academic Press, 1977), pp.

39-50.

14. Aunstrup, K., Andresen, O., Falch, E. Z,, and Mielsen, T. K., “Production of Microbial Enzymes, ” in

Microbial Technolo~: Microbial Processes, vol. 1,

H. J. Peppier and D. Perlman (eds.) (New York:

Academic Press, 1979), pp. 282-311.

15. Bernard Wolnak & Associates, in Food Prod., July

1978, p. 41.

16. Solomons, G. L., “The Microbial Production of En17

18

19

20.

zymes, ” in Biotechnological App/ica/ions of Proteins and Enzymes, Z. Bohak and N. Sharon (eds.)

(New York: Academic Press, 1977), pp. 51-61.

Chemical and Engineering News, Apr. 14, 1980, p.

6.

Levine, W. G., “Anticoagulants, Antithrombotic,

and Thrombolytic Drugs, ” in The Pharmacological

Basis of Therapeutics, 5th cd., L. S. Goodman and

A. Gilman (:eds.) (New York: MacMillan Publishing Co., Inc., 1975), p. 1,365.

Gilman, A. G., and Muriad, F,, “Androgens and

Anabolic Steroids, ” in The Pharmacological Basis

of Therapeutics, 5th cd., L. S. Goodman and A.

Gilman (eds.) (New York: MacMillan Publishing

Co., Inc., 1975), pp. 1,451-1,471.

Gilman A. G., an-d- Muriad, F., “Estrogens and Progestins, ” in The Pharmacological Basis of Therapeutics, 5th cd., L. S. Goodman and A. C1ilman

(eds.) (New York: MacMillan Publishing Co., Inc.,

1975), pp. 1,423-1,450.

21. Haynes, R. C., and Lamer, J., “Adrenocorticotropic Hormone: Adrenocorticol Steroids and

Their Synthetic Analogs: inhibitors of Adrenocorticol Steroid Biosynthesis, ” in The Pharmacological Basis of Therapeutics, 5th cd., L. S, Cmodman and A. Gilman (eds. ) (New York: MacMillan

Publishing Co., Inc., 1975), pp. 1,472-1,506.

22. Sebek, O. K., and Perlman, D., “Microbial Transformation of Steroids and Sterols,” in Microbial

Technology: Microbial Processes, vol. 1, H. J. Peppier and D. Perlman (eds.) (New York: Academic

Press, 1979), pp. 483-496.

23. Brazeau P., “Agents Affecting the Renal Conser-
Appendix l-B—A Tjmetable for the Commercial Production of Compounds ● 291

24

25

l~ation of Water, “ in The Pharmacological Basis of

Therapeutics, 5th cd., L. S. Goodman and A.

Gilman (eds.) (New York: MacMillan Publishing

CO., IIw., 1975), PP. 858-859.

Haynes, R. C., and Lamer, J., “Insulin and Oral

Hypogl.vcemic Drugs: Glucagon,” in The Pharmacological Basis of Therapeutics, 5th cd., L. S. Cmodman and A. Gilman (eds. ) (New York: MacMi]]an

Publishing Co., Inc., 1975), pp. 1,507-1)533.

Jawetz, E., Melnick, J. L., and Adelberg, E. Z., Reviews of Medical A4icrobioloqv, 1 Ith” cd., (Los

Altos, Ca]if.: Lange Medical Publications, 1974).

26. Thaler, M. M., Biochem. Bioph-vs. f?es. Comm.

54:562, 1973.

27. Nakiio, Y., “hficrobia] Production of Nucleosides

28,

29<

and Nuckwt ides, ” in Microbial Technology.. Microbial Processes, twl. 1, H. J. Peppier and D. Perlman

(eds.) (Newf York: Academic Press, 1979), pp.

312.3,55.

Perlman, D., “Microbial Production of Antibiotics, “ in Microbial Technology Microbial Processes,

\rol. 1, H. J. Peppier and D. Perlman (eds. ) (New

York: Academic Press, 1979), pp. 241-281.

Weatherall, D. J., and Ciegg, J. B., “Recent De\’elopments in the Molecular (;enetics of Human

Hemoglobin,” Cell 16:467-480, 1979.

30. Ignoffo, C. M., and Anderson, R. F., “Bioinsecticides, ” in Microbial Technology: Microbial Processes, vol. 1, H. J. Peppier and D. Perlman (eds.)

(New York: Academic Press, 1979), pp. 211-.z4o.

31. German, M., and Huber, F. M., “@Lactam Antibiotics, ” in Annua[ Reports on Fermentation Processes, vol. 2, G. T. Tsao and D. Perlman (eds. )

32,

33.

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35.

(New York: Academic Press, 1978), pp. 203-222.

Shibata, M., and [JLVeda, hf., “Microbial Transformation of Antibiotics, ” in Annuai Reports on

Fermentation Processes, Yrol. 2, G. T. Tsao and D.

Perlman (eds.) (New York: Academic Press, 1978),

pp. 267-304.

Lockwood, L. B., “Production of Organic Acids by

Fermentation, ” in Microbial Technology: Microbial

Processes, vol. 1, H. J. Peppier and D. Per]man

(eds.) (New York: Academic pl’ess, 1979), pp.

367-372.

Roberts, J. D., and Caserio, M. C., Basic Principles

oj’ Organic Chemistry (New York: Benjamin, Inc.,

1964), p. 1,096.

tVindholz, M. (cd.), The Merck Inde,x, 9th ed. (Rahway, N. J.: Merck & Co., 1976).
Appendix I-C

Chemical and Biological Processes

A comparison was made of waste stream pollution

for chemical and biological processes. Ideally, the

comparison should be between the two processes

used in the production of the same end product.

Since such data do not currently exist at the industrial level, the comparison was made between the

chemical production of a mixture of chemicals and

the biological production of alcohol and antibiotics.

one noteworthy parameter is the 5-day biochemical

oxygen demand (BOD5) —the oxygen required over a

5-day period by organisms that consume degradable

organics in the waste stream. If the oxygen demand

is too high, the discharge of such a stream into a

body of water will deplete the dissolved oxygen to

the point that it threatens aquatic life. An important

variable that must be considered along with the BOD

is the COD (the chemical oxygen demand). Large differences between the COD and BOD of a waste system can indicate the presence of nonbiodegradable

substances. Although the conventional process

stream shown in table 1-C-1 has less BOD5 than the

biological process stream, its COD content probably

means that non biodegradables are present, and

specialized waste treatment is necessary.

BOD is one area where traditional fermentation

based processes have posed pollution problems.

Batch fermentation processes typically generate

large quantities of dead cells and residual nutrients

that cause a large BOD if they are dumped directly

into a dynamic aquatic environment. (See table I-C-1.)

This difficulty can be circumvented by the use of

spent cell material as an animal feed supplement or

Table I-C-1.— Waste Stream Pollution Parameters:

Current Processes v. Biological Processes

Compounds: Mixed chemicals, including ethylene

oxide, propylene oxide, glycols, amines, and ethers

Current Biological

Pollution parameters processes processes

Alkalinity (mg/l).

BOD5

a

(mg/l). . .

Chlorides (mg/l)

C O D

b

(mg/l). . . .

Oils (mg/l) . . . . .

pH . . . . . . . . . . .

Sulfates (mg/l) .

. . . . . . . . . . 4,060 0

. . . . . . . . . . 1,950 4,000-12,000

. . . . . . . . . . 430-800 0

. . . . . . . . . . 7,970-8540 5,000-13,000

. . . . . . . . . . 547 0

. . . . . . . . . . 9.4-9.8 4-7

. . . . . . . . . . 655 0

Total nitrogen (mg/l). . . . . . . 1,160-1,253 50-200

Phosphates (mg/l) . . . . . . . . 0 50-200

as.day biOk@Cal oxygen demand.

bchemical oxygen demand.

SOURCE: Office of Technology Assessment.

as a fertilizer. These applications have been intensively investigated and have met with success.

Because of the renewed interest generated by the

potentials of genetic engineering, some traditional

fermentation systems are being redesigned. Immobilization allows the reuse of cells that would otherwise be discarded. These systems can be used continuously for several months as compared with the

usual fermentation time in a batch process of about

one week or less. [remobilized operations create

waste cells much less often than batch systems, and

therefore generate less BOD.

292
Appendix I-D

The Impact of Genetics on Ethanol—

A Case Study

Objective

This study examines how genetics can and will affect the utilization of biomass for liquid fuels production. There are two major areas where genetics are

applicable. One is in plant breeding to improve availability (both quantity and quality) of biomass resources (with existing and previously unused land);

the second is in the application of both classical

mutation and selection procedures and the new genetic engineering techniques to develop more efficient microbial strains for biomass conversion. Examples of goals in a plant breeding program would

include improvements in photosynthetic efficiencies,

increased carbohydrate content, decreased or modified lignin content, adaptation of high productivity

Figure l-D-l .—An Overview of Alternative

plants to poor quality land, improved disease resistance, and so forth. However, the focus here is entirely on the second area, the use of genetics to improve

microbial-based conversion to produce ethanol.

In order to assess the type and extent of improvements in micro-organisms that might benefit

ethanol production, its process technology and

economics must first be examined. An overview of

the biomass conversion technology is presented in

figure I-D-1; processes are defined mainly on the

basis of the primary raw material and the type of

pretreatment required to produce mono- or disaccharides prior to fermentation. In addition, there

are several alternative fermentation routes to produce ethanol; these are characterized by the type of

micro-organisms and will be examined with the inRoutes for Conversion of Biomass to Ethanol

SOURCE: Massachusetts Institute of Technology.

76-565 0 - 81 - 20

293
294 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

tent of quantifying the potential impact of genetic improvement on each one. It is interesting to note that

each type of organism has its substrate restrictions, .

and only the anaerobic bacteria such as Clostridium

thermosaccharolyticum and C. thermohydrosulforicum can utilize all of the available substrate.

Substrate pretreatment

Pretreatment refers to the processing that is required to convert a raw material such as sugarcane,

starch, or cellulosic biomass to a product that is

fermentable to ethanol. In most cases, the pretreatment is either extraction of a sugar or hydrolysis of a

polysaccharide to yield a mono-or disaccharide.

EXTRACTION OF SUGAR

Sugar crops such as sugarcane, sugar beets, or

sweet sorghum are highly desirable raw materials

for producing ethanol. These crops contain high

amounts of sugars as sucrose. In addition, the yield

of fermentable material per acre is high; sugarcane

and sugar beets yield 7.5 and 4.1 dry tons of biomass

per acre, respectively. ’

Sugar is extracted from cane or beets with hot

water and then recrystallized. The resulting sugars

are utilized directly by organisms having invertase

activity (to split sucrose to glucose plus fructose).

Molasses, a sugary byproduct of the crystallization of

sucrose, may also contain sucrose although in most

cases it is inverted with acid.

The primary use for sugar crops is food sugar.

Sugar sells for over 20 cents/lb. Molasses, which currently sells for about $100/ton (about 10 cents/lb

sugar) is used extensively as an animal feed. Substantial amounts of both sugar and molasses are imported into the United States for food uses and are

therefore unavailable for ethanol production, There

are proposals to increase sugar production for use as

an energy crop; however, this will require the

development of new land for sugar production.

STARCH

The primary raw material for ethanol fermentation in the United States is cornstarch. Corn processed by wet milling, yields about 36 lb of starch

from each 56 lb bu; this amount of starch will produce 2.5 gal of absolute ethanol. Corn yields are

typically 80 to 120 bu/acre so that 200 to 300 gal of

ethanol can be derived per acre of corn per year.

Pretreatment of starch is initiated by a gelatinization step whereby a starch slurry is heated for 5 min

at 105° C. After cooling to 98° C, œ-amylase is added

‘Paul B. Weisz and John F. Marshall, Science 206:24, 1979.

to break down the starch to about 15DE (dextrose

equivalents). This process of liquefaction reduces the

viscosity such that the solution can be easily mixed.

After further cooling to 30° C, glucoamylase is added

along with a starting culture of yeast so that saccharification and fermentation proceed simultaneously.

The resulting fermentation, to produce typically 8 to

10 percent ethanol (volume per volume), requires 42

to 48 hr for completion. This compares with a 16- to

20-hr fermentation if sugar as molasses or cane juice

is used as the substrate. Thus, the use of starch requires the addition of enzymes prior to and during

fermentation, as well as large fermenter capacity as a

consequence of the slower fermentation time compared with sugar substrates.

Improvement in the economy of ethanol fermentation based on starch is possible by developing a

micro-organism that can produce wamylase and

glucoamylase and thus eliminate the need to add

these enzymes. Since the rate of fermentation depends on the rate of starch hydrolysis, increased levels of glucoamylase may enhance the rate of starch

hydrolysis and thus increase the rate of ethanol production, This would lower the capital requirements

as well as the cost of enzyme addition.

CELLULOSIC BIOMASS

Processes for the utilization of cellulosic biomass

to produce liquid fuels all have three features in common:

1. They employ some means of pretreatment to at

2.

3.

least effect some initial size reduction and, more

often, cause a disassociation of lignin and cellulose;

they involve either acid or enzymatic hydrolysis

of the cellulose and hemicellulose to produce

mono- and disaccharides; and

they employ fermentation to produce ethanol or

some other chemical.

A wide variety of process schemes have been proposed for the conversion of cellulosic biomass to

liquid fuels; a summary of the major steps in two

acid hydrolysis and three enzymatic hydrolysis

schemes in shown in figures I-D-2 and I-D-3. The initial size reduction is required to increase the

amount of biomass surface area that can be contacted with acid, solvent, steam, enzymes, or

chemicals that might be used to disassociate the

cellulose and hemicellulose from the lignin.

Pretreatments that have been investigated to

facilitate the process are summarized in table

I-D-1. The problems with pretreatment are that they

require energy, equipment, and often chemicals;

they result in an irretrievable loss of sugar, and in

undesirable side-reactions and byproduct forma-
Appendix l-D—The Impact of Genetics on Ethanol—A Case Study ● 295

Figure l-D-2.–Alternative Schemes for Acid Hydrolysis of Cellulosic Biomass for Ethanol Production

SOURCE: Massachusetts Institute of Technology.

tion. Furthermore, if acids, alkali, or organic chemicals are used, they must be recycled to minimize

cost or disposed of in order to prevent pollution.

In starch processing, prior to ethanol fermentation, mechanical grinding, steam, and enzymes are

employed. The energy requirements are small and

contribute relatively little to the final ethanol cost.

The objective in the development of cellulose-based

processes should be to minimize both energy and

chemical requirements. The development and scaleup of effective pretreatment technology are under

active investigation and require continued financial

support to better develop several alternative routes.

The most promising routes are: steam treatment, solvent delignification, dilute acid, cellulose dissolution,

and direct fermentation.

Several different acid hydrolysis schemes have

been proposed. However, most appear as in flow

scheme A or B in figure I-D-2. Dilute acid is used to

hydrolyze the hemicellulose to pentose sugars primarily and then stronger acid at higher tempera-

‘Proceedings of 3rd Annual Biomass Energv System Conference, National

Technical Information Set-vice, SfXl~rP-33-285, 1979.

tures is used to cause cellulose hydrolysis (scheme

A). A major problem with this approach is the irreversible loss of sugars to undesirable side-product

formation. After separation of residual solids (mostly

lignin), which can be burned to provide energy for

distillation, the sugar solution is fermented by yeast

to ethanol. The pentose sugars also can be fermented, but by organisms other than the ethanol

producing yeast, to other chemicals, some of which

could be used as fuels (e.g., ethanol, acetic acid,

acetone, butanol, 2,3-butanediol, etc.).

An alternative (scheme B, figure I-D-2) to the above

is to use a solvent, after pentose sugar removal, to

dissolve the cellulose, allowing its separation from

lignin. This cellulose solution is easily and efficiently

hydrolyzed to sugars. The advantage of this approach over the direct acid hydrolysis is that the

yield of sugar is much higher. In the harsh acid hydrolysis, considerable sugar is destroyed. However,

the major disadvantage of both these schemes is that

they require recycling or disposal of acids and

solvents. A second problem is that almost nothing is

known about how to scale-up some of the newly de-
296 . Impacts of Applied Genetics —Micro-Organisms, Plants, and Animals

Figure I-D-3.—Alternative Schemes for Enzymatic Hydrolysis of Cellulosic Biomass for Ethanol Production

SOURCE: Massachusetts Institute of Technology.

Table I- D-1 .-Alternative Pretreatment Methods for

Lignocellulose Materials

Chemical methods Physical methods

Sodium hydroxide (alkali) Steam

Ammonia Grinding and milling

Chemical pulping Irradiation

Ammonium bisulfite Freezing

Sulfite

Sodium chlorite

Organic solvents

Acids

SOURCE: Office of Technology Assessment.

veloped technology, such as that developed by

groups working at Purdue University, New York University, and Dartmouth College. There are several

engineering problems involving both heat and mass

transfer and acid/solvent recycle that need to be evaluated at larger scale. At least some of this work will

be done at the process development unit now being

built at the Georgia Institute of Technology. The

most promising directions that need development

are:

● the scale-up of high rates and high yield laboratory hydrolysis systems, and

● the development of methods for acid and chemical recycle schemes.

There are three types of approaches that have

been employed for enzymatic hydrolysis of cellulosic

biomass. These are summarized in figure I-D-3. They

all involve some initial size reduction to increase the

surface area available for enzymatic attack. In

schemes A and B, the incoming cellulosic biomass is

split into two streams; one is used to grow organisms

that produce cellulolytic enzymes called cellulases,

and the other is used to produce sugar.

In scheme A, the cellulases are recovered and then

added to a separate enzyme hydrolysis reaction.

They hydrolyze both the cellulose and hemicellulose,

and the resulting sugar solution is then passed to an

ethanol fermentation stage where hexoses are converted by a yeast fermentation to ethanol. Utilization

of the pentose requires a separate fermentation. Re-
Appendix l-D—The Impact of Genetics on Ethanol—A Case Study ● 297

sidual lignin, which is removed before (by solvents

extraction) or after hydrolysis, is used to provide

energy for ethanol recovery, Extensive work on this

approach has been done at the University of California, Berkeley, and the U.S. Army Natick Laboratories.

In scheme B, the cellulase is not recovered but

rather, the whole fermentation broth from cellulase

production is added to the cellulosic biomass along

with ethanol-producing yeast. The result is a simultaneous cellulose hydrolysis (saccharification) and

fermentation. (In the production of ethanol from

starch, the starch hydrolyzing enzymes are added at

the same time as the yeast for simultaneous saccharification and fermentation.) This technology has been

demonstrated by the Gulf Oil Co. After fermentation,

the ethanol is recovered and the residual lignin can

again be used for energy for distillation. The problem of unused pentose sugar still remains and will require a separate fermentation step.

A third alternative (scheme C, figure I-D-3) shows a

simpler approach, namely a direct fermentation on

cellulose. This approach has been developed at the

Massachusetts Institute of Technology. It utilizes

bacteria that will produce cellulase to hydrolyze the

cellulose and hemicellulose and ferment both the

hexose and pentose sugars to ethanol in a singlestage reactor. The advantage of this approach is a

minimal requirement for pretreatment, a combined

enzyme production, cellulose hydrolysis and ethanol

fermentation, and simultaneous conversion of both

pentose and hexose sugars to ethanol. This concept

is new and work still needs to be done to increase the

ethanol concentration, minimize side product formation, and increase the rate of ethanol production.

Again, residual lignin will be used to provide the

energy for ethanol distillation.

FERMENTATION OF ETHANOL

An examination of the economics for ethanol production shows that the dominant cost is the process

raw material. As seen in table 1-D-2 the feedstock represents 60 to 70 percent of the manufacturing cost.

Thus, it is clear that any improvement in substrate

utilization efficiency is of substantial benefit. The

theoretical yields of ethanol from glucose, sucrose,

and starch or cellulose are 0.51, 0.54 and 0.57 gram

(g) ethanol/g material, respectively; the differences

result from the addition of a molecule of water on

hydrolysis. There are several approaches to improve

the yield above the typical value of 90 to 95 percent

currentl y achieved. These are:

. increase the ratio of ethanol produced per unit

weight of cells, e.g., through cell recycle,

vacuum fermentation, immobilized cells, or improvement in specific productivity (g ethanol/g

m

Table I-D-2.—A Comparison of the Distribution of

Manufacturing Costs for Several Ethanol

Production Processes

Grain

Substrate Molasses Corn Sorghum

Cost component (%)

Capital. . . . . . . . . . . . . . . . 9

Operating . . . . . . . . . . . . . 20

Feedstock. . . . . . . . . . . . . 71

Total . . . . . . . . . . . . . . . 100

Cost on energy basis

($MMBtu) . . . . . . . . . . . . . 12.5

Cost/gal ethanol ($/gal) . . . . 1.05

Capital investment

($/annual gal) . . . . . . . . . . 1.02

12

26

62

10

30

60

100

14.9

1.25

1.05

100

12.7

1.07

1.75

SOURCE: “Comparative Economic Assessment of Ethanol From Biomass,”

Mitre Corp., report HCP/ET-2854).

cell hr), by increasing the content and/or activity of those enzymes in the pathway to ethanol;

increase the utilization of other materials in the

substrate, e.g., the use of oligosaccharides, especially branched, in starch, and the use of contaminating sugars such as galactose or mannose

for hemicellulose; and

develop a route for the utilization of pentose sugars, especially xylose, present in hemicellulose.

The potential effect of oligosaccharides or contaminating sugar utilization is relatively small, since

they represent typically 1 to 3 percent of the total

sugar content. However, if cellulosic biomass containing 15 to 25 percent hemicellulose is used, then

the impact of pentose conversion to ethanol is great.

Cellulosic biomass is made up primarily of cellulose, hemicellulose (mostly xylan) and lignin. Other

components such as protein, ash, fats, etc., typically

comprise about 10 percent. The composition of biomass can be expressed in terms of the following

equation:

F,. + F,, + F,, = 10 [1]

1 – 1 \

where FC, FH, F~) and FA are the weight fractions of

cellulose, hemicellulose, lignin, and ash, respectively.

Assuming that the ash is 10 percent (FA = 0.1) and

that FC and FH are the only fermentable components

in the biomass, then:

F C = FH = 0.9 – F L (2)

The maximum amount of ethanol from one unit of

biomass (Y~~,~) is;

YWCFC = YE/HFH = Y~jB (3)

Where Yi,,C and Yk;lH are the yield of ethanol for cellulose and hemicellulose, respectively. Equation 2

can be rearranged to relate the fractions of cellulose:
298 . Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

Fc = 0.9 – F L - FH (4)

Substituting this into equation 3 gives:

YE/B + YE/c(0.9 – FL – FH) + YE/HFH (5)

From equation 5, the effect can be calculated of

hemicellulose content and conversion yield on the

overall conversion of biomass to ethanol. Assuming a

lignin content of 15 percent (FL = 0.15) and using YE/c

= 0.57 g/g the following equation is obtained:

Y E / B

= 0.43 + F H(YE/H - 0.57) (6)

The theoretical yield value on hemicellulose,

YE/H, is not well-defined because so little is known

about the biochemistry of anaerobic pentose

metabolism. If one mole of ethanol is produced per

mole of xylose, the yield is 0.3 g ethanol/g xylose. It

two moles of ethanol could be obained, YE/H would

be 0.61; however, neither the mechanism nor the

thermodynamics of the conversion is sufficiently

well-defined to allow one to expect this value. The

maximum observed values are about 0.41 g

ethanol/g xylose.3 The sensitivity of the overall

yield to this value is shown in figure I-D-4. The impact of pentose utilization depends on the amount

%. D. Wang and [:. Cooney, Massachusetts Institute of Technolo~v, unpublished results.

Figure l= D-4.-Effect of Pentose Yieid (YE/H) on

Overall Yield of Ethanol from Cellulosic Biomass

(YE/B) with Varying Fractions of Hemicellulose (FH).

0.5

0 0.2 0.4 0.6

Yield on pentose (g ethanol/g pentose)

SOURCE: Massachusetts Institute of Technology.

of hemicellulose present. From the value in figure

I-D-4 and the observation that 70 percent of the

manufacturing cost is the raw material cost, it is

possible to estimate the economic benefit of pentose utilization. Equation 7 relates the overall

ethanol yield to the manufacturing cost:

C E = CB x 6 . 6

(7)

Y E / B

().7

where CE is the manufacturing cost per gallon of

ethanol, CB is biomass cost (cents/lb), 6.6 is the conversion from pound to gallon of ethanol, and 0.7 is

the 70-percent factor for relative biomass cost to

ethanol cost. For a biomass costing 2 cents/lb and

containing 20 percent hemicellulose, the manufacturing cost is reduced from 59 to 43 cents/gal, when

the yield on pentose goes from zero to 0.6.

At the present time, there are few organisms that

produce more than one mole of ethanol per mole of

pentose and none of the usual alcohol producing

yeasts will ferment pentoses to ethanol. Addition or

improvement of the ability to use pentose will have a

major impact on the economics of ethanol production.

The second major cost in ethanol production relates to the cost of operation. Typically, 20 to 30 percent of the final manufacturing cost is accounted for

by the sum of labor, plant overhead, administration,

chemical supplies, and fuel costs. The chemical supplies represent less than 1 cent/gal ethanol and may

be neglected. The labor, overhead, and marketing

costs vary with plant size, but represent 11 to 7

cents/gal for a 20 to 100 million gal/yr plant, respectively. Any improvement in the reduction of plant

size or complexity will reduce this cost; however, the

economic impact is small. The major component of

the operating cost is the fuel charge for plant operation and for distillation. Plant operations, e.g., mixing, pumping, sterilization, starch gelatinization,

biomass grinding, etc., represent about 20 to 30 percent of the energy cost. The remainder is for ethanol

distillation and residual solids drying. Considerable

effort has been focused on methods to improve the

energy efficiency, of distillation to reduce it from the

160)000 Btu/gal required for beverage alcohol. While

considerable differences in opinion exist as to the

minimum, a reasonable expectation is about 40,000

Btu/gal although current technology requires 69,000

Btu/gal. 4 Forty thousand Btu is about half of the energy content of ethanol per gallon.

A discussion of process improvements relating to

ethanol recovery has two components. The first is

4Report of (he Gasohol Stud-y C,roup of the Energy Research Advisory

Soard, Department of Energv, Washington, D. C., 1980. ‘M. Gibbs, and R,

D. DeMoss, “Ethanol Formation in Pseudomonas /indneri, ” Arc/I. Biochem

Biophys. 34:478-479, 1951.
Appendix l-D—The Impact of Genetics on Ethanol—A Case Study . 299

related to operating costs and the second is related to

energy efficiency. If coal is used to provide energy

for distillation, and it is valued at $30/ton, with

10,500 Btu/lb or $1.50/million Btu, then the energy

cost for distillation (optimistically assuming 40,000

Btu/gal) is $6/gal. If lignin from cellulosic biomass is

used as a fuel, the cost is reduced further. On the

other hand, if oil at $40/bbl (130,000 Btu/gal and 42

gal/bbl) or $7/million Btu is used, then the energy

cost is 28 cents/gal of ethanol.

From a common sense, economic, and political

point of view, it does not seem reasonable to utilize

liquid fuel to produce liquid fuel from biomass.

Therefore, it will be assumed that petroleum will not

be used for distillation and that either coal or biomass will be employed.

In order to assess the impact of process improvements on the energy demand, it is necessary to look

at an overall material balance. This is summarized in

figure 1-D-5. Only a portion of the entering biomass

feedstock is fermented to ethanol and there are two

product streams, one containing ethanol and the

other solids, both must be separated from water. It is

important to note that as the ethanol concentration is

increased, the energy requirement for both ethanol

recovery from the water and for drying will decrease. Therefore, the impact of developing ethanol

tolerant micro-organisms is seen as a reduction in

energy cost.

Figure l-D-5. - Process Schematic for Material and

Energy Balance

Biomass

Process heat

Ethanol

Solids

SOURCE: Massachusetts Institute of Technology.

The third major cost for ethanol manufacturing is

the capital investment, which represents about 4 to

12 percent of the manufacturing cost. The capital investment is determined by the complexity of the

processes and the volumetric productivity of ethanol

production. Thus, the development of a micro-organism that will require a minimum amount of feedstock pretreatment and will produce ethanol at a

higher rate will reduce the net capital investment.

The volumetric productivity (QE

) for ethanol production is given by:

QE

= qpX

where qP is the specific productivity expressed in g

ethanol per g cell hr, and X is the culture density.

Therefore, there are two approaches to obtain high

productivity: first, to choose or create an organism

with a high specific rate of ethanol production and

second, to design a process with high cell density.

The application of genetics can be used to enhance

the intracellular enzyme activity of the enzymes

used for ethanol production. The resulting increase

in qp

, will result in reduced capital investment requirements.

There are four types of ethanol processes based

on different organisms; they are:

1. Saccharomyces cerevisiae and related yeast,

2. Saccharomyces cerevisiae/Trichoderma reesei,

3. Zymomonas mobilis, and

4. Clostridium thermocellum/thermosaccharolyticum, or thermohydrosulfuricum.

The first is the traditional yeast based process using

S. cerevisiae to ferment soluble hexose sugar to ethanol. In the second, the substrate range is extended

to cellulose by the use of cellulase produced by T.

reesei. The third approach utilizes Z. mobilis; this

organism is a particularly fast and high ethanol yielding one. Its range of fermentable substrates, however, is limited to soluble hexose sugars.

In many tropical areas of the Americas, Africa,

and Asia, alcoholic beverages prepared from a mixed

fermentation of plant steeps are popular. Bacteria

from the genus Zymomonas are commonly employed. In the early 1950’s, the genus Zymononas acquired a certain fame among biochemists by the discovery that the anaerobic catabolism of glucose

follows the Enter-Doudoroff mechanism.’ This was

very surprising, since Zymomonas was the first example of an anaerobic organism using a pathway

mainly in strictly aerobic bateria.6

In spite of its extensive use in many parts of the

world, its great social implications as an ethanol pro-

‘M. Gibbs and R. D, de Moss, “Ethanol Formation, in Psuedornonas

Iindneri,” Arch. Biochem. Biophys., 34;478-$79, 1951.

6J. Swings and J. Del.ey, “-rhe Biologv of Zymomonas, ” Bacteriological Reviews 41: 1-46, 1977.
300 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

ducer, and its unique biochemical position, Zymomonas has not been studied extensively. T

The organism most often studied is Zymomonas

mobilis, which can produce up to 1.9 moles of

ethanol per mole of glucose. Recent studies reported

from Australia, have established the Z. mobilis can

ferment high concentrations of glucose rapidly to

ethanol in both batch and continuous culture with

higher specific glucose uptakes rates for glucose and

ethanol production rates than for yeasts currently

used in alcohol fermentations in Australian.89

For example, several kinetic parameters for a Z.

mobilis fermentation were compared with Saccharomyces carlsbergensis10 specially selected for its

sugar and alcohol tolerance.ll Both specific ethanol

productivity and specific glucose uptake rate are several times greater for Z. mobilis. This result is mainly

due to lower levels of biomass formation and glucose

consumption. The lower biomass produced would

seem to be a consequence of the lower energy available for growth with Zymomonas than with yeasts—

the Enter-Doudoroff pathway producing only 1 mole

of adenosine triphosphate (ATP) per mole of glucose,

compared to glycolysis with 2 moles ATP per mole

glucose. In none of the first three examples can ethanol be produced from pentose sugar.

The fourth approach utilizes a mixed culture of

Clostridia, which will utilize cellulose and hemicellulose, hexoses, and pentoses for ethanol production.

The application of genetics for

improving microbial strains

In the previous sections, the process steps have

been identified that are particularly sensitive to the

quality of the microbial strains. The following are improvements of microbial characteristics that are

either now possible or might be so in the future and

that will have an impact on the overall economics of

the process. The effect of new genetic techniques requiring future research is similar for all micro-organisms in two ways.

1. Manipulations could be attempted today with

less effort and greater chance of success if tools

like cell fusion and recombinant DNA (rDNA)

techniques were available for all of the microbes of interest.

——

T;ibbs, et al., op. cit.

‘K. J. Lee, D. E. Tribe, and P. L. Rogers, “Ethanol Production by Zymomonas mobilis in Continuous Culture at High Glucose Concentrations,” Biotechnology Lett. 421-426, 1979.

9P. L. Rogers, K. J. ke, and D. E. Tribe, Biotechnoi. Lett. 1:165-170, 1979.

IOlbid.

I ID. Rose, Proc. Bichem. 11 [2), 1976, pp. IO-12.

2. Manipulations require further knowledge in a

specific area or the development of an entirely

new genetic system in ethanol producing microbes—e.g., there is no genetic system for the

thermophilic anaerobic bacteria. Knowledge on

how to genetically alter ethanol tolerance of

both bacteria and yeast is lacking.

The economics of the fermentation of a substrate

into alcohol is primarily controlled by three factors:

1.

2.

3.

Ethanol yield.—The amount of product produced per unit of substrate determines the major raw materials cost of the fermentation.

Final ethanol concentration.—The cost of separating the ethanol from the fermentation broth is a

function of the ethanol concentration in that

broth.

Productivity. —The amount of ethanol produced

per liter of fermenter volume per hour determines the capital cost of the fermentation step,

once the type of fermenter and the annual output have been chosen. Productivity is not independent of the final ethanol concentration, and

so an optimum compromise between these variables must be chosen.

The impact of genetics on ethanol yield

Most microbes that are chosen for making ethanol

already produce nearly the theoretical maximum

yield. In these cases little improvement can be made.

The yield may be lower when the microbe has

been chosen for its other technical advantages such

as ability to degrade cellulose. Lower yield of a

microbial end product, like ethanol, can result from

the diversion of substrate to cell mass or to an alternative product. Both of these faults can be readily attacked. A number of cell changes (e.g., leaky membranes) can cause the microbe to waste energy, requiring it to metabolize more substrate into alcohol

to make the same cell mass. Where the thermodynamics and redox balance of the fermentation

allow, unwanted waste products can be eliminated

by mutation of the relevant pathways. Only limited

work has been done on this type of research with industrially significant bacteria.

The impact of genetics on final ethanol

concentration

This is amenable to genetic manipulation, both empirical and planned. An improvement in ethanol tolerance decreased both separation costs and fermenter capital cost (through increased productivity).

When traditional distillation is used, the effect on
Appendix I-D—The Impact of Genetics on Ethanol—A Case Study ● 301

the separation cost of increased ethanol tolerance is

smaller once ethanol concentrations have reached

approximately 6 percent. However, the importance

of increased ethanol concentration to fermenter productivity remains.

It is likely that the most important inhibitory action of ethanol takes place at the cell membrane.

Strategies for manipulating the cell membrane composition and properties, and understanding in this

area, are increasing rapidly.

Genetics and ethanol tolerance

The study of ethanol tolerance by micro-organisms has been approached using strains with

altered genetic makeup. Several kinds of Escherichia

coli mutants have been isolated having different

tolerances to ethyl alcohol. 12 Solvent resistant strains

either had larger amounts of total phospholipid (type

III) or had an altered phospholipid and membranebound protein composition (type 11). On the other

hand, mutants with a lesion mapping close to pss

gene (which codes for phosphotidylserine synthetase) were either solvent sensitive or resistant .13

The physiology of an E. coli ethanol resistant mutant has been characterized similarly .14 This strain

had pleiotropic growth defects including abnormal

cell division and morphology. It also had an altered

lac permease that was not due to a mutation in the Y

gene. It was concluded that altered membrane composition was responsible for this abnormal behavior.

More recently, ethanol tolerant mutants have been

isolated from C. thermocellum. 15 Indirect evidence

lead to the conclusion that strain S-4 was defective in

hydrogenate, since this strain produced lower

amounts of acetic acid.l6 A different ethanol resistant

isolate of the same bacterium, strain C9, proved to

have a lower activation energy for growth than the

wild type, a property that has been related to membrane composition.

There are three categories of changes that could

influence the fermentation process:

1. Manipulate the existing controls on metabolism.

Consider an example. In many organisms the

IZI), p. (;l~rk and J, P, Beard, “Aitered phospholipid Composition in Mutants

of’ Escherichia Co/i %msitive or Resistant to organic Solvents, ” J. Gen.

Mirrobio/. 1 13:267-274, 1979.

IJA. ohta and I. Shibuva, “Membrane Phospho]ipid Synthesis and Phenot,vpic (correlation of an ~. Co/ipss Mutant,” J. Bacferio/. 132:434443, 1977.

14L, ,,~, ~,l.ied ~lld A, N~\,i(:k, “organic Soh,ents as Probes for the structure

iind k’unrtion of the Bacterial Membrane: Effects of Ethanol on the Wild

Type and as F,thanol Resistant hfutant of’ Fscherichia Co/i, ” J. Bacteriol.

114:239-248, 1973

‘%. D. Wang, ‘production of P;thanol From (;ellulose by [,loslridiurn Ther.

mocellum, ” M.S. Thesis, Department of Nutrition and Food Science, ,Massa.

chusetts Institute of Technology, ]979.

Idbid.

energy level of the cell, expressed through

adenosine monophosphate (AMP), adenosine diphosphate (ADP), and adenosine triphosphate

(ATP) levels, partially controls the rate of glycolysis. A defective cell membrane would provide an energy sink, to keep glycolysis at its

maximum rate. Strategies such as this could be

attempted now.

2. Increase the amount of each transport and catabolic enzyme in the fermentation pathway. This

requires the ability to isolate the genes of interest and to amplify them with in vivo or in

vitro recombinant techniques in the microbe of

interest. This is not an immediate prospect.

3. Accomplish complete deregulation of the fermentation pathway in the microbe of interest.

Essential catabolic enzymes are difficult to

manipulate, and this is also not an immediate

prospect.

Genetic manipulation of the microbe can influence

fermentation processes in other ways as well. These

are less important than improvements in yield, final

ethanol concentration, and productivity, but they

also affect the cost. Examples are:

type of fermenter used;

nonsubstrate nutrients;

strain stability;

cell separations for byproducts, recycle, or ethanol recovery (i.e., increased size for recovery);

operating conditions, i.e., higher growth temperatures for yeast and mesophilic bacteria; and

range and efficiency of substrate utilization (i.e.,

complete utilization of all sugars).

More detailed examples are:

● Type of fermenter. —If the organism, whether it

be a yeast or a bacterium, can be made to grow

under conditions of pH, ethanol concentration, temperature, etc., that preclude contamination, inexpensive lined basins can be used instead of tanks, since

steam sterilization of the fermenter is not required.

In this case, some operating and capital costs associated with sterilization are avoided as well.

A type of continuous beer fermenter requires

growth in the form of fast-settling pellets. In other

fermenters, fast-settling particles (such as mycelia)

present problems that are best avoided by agglomeration of the cell mass. This type of control over the

growth form of micro-organisms is amenable to

genetic manipulations.

● Nonsubstrate medium costs. —In addition to the

carbon-energy substrate and water, growing cells

must be supplied with other nutrients. Some organisms can make all of their biochemical from quite

simple sources of nitrogen, phosphorus, sulfur,

magnesium and trace metals. Others require more
302 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

complex molecules, ready-made, such as amino acids

and vitamins.

The more cheaply these nutrient needs can be

provided, the better. Whenever an organism can be

given genes from another source by applied biotechnology techniques, there is a possibility that complex

nutrient requirements can be obviated. However,

this requires that all the genes in a given pathway be

located in the source and be made to function in the

new microbes. The feasibility of this is uncertain, but

solutions would decrease the cost of producing ethanol with yeast as well as clostridia.

● Stain stability.—Many of the suggested ethanol

processes propose to employ continuous culture.

Although this offers several advantages over batch

culture, it is somewhat vulnerable to deleterious mutations of the microbe used, particularly if the microbe has been extensively altered in ways that make

it less competitive.

These deleterious genetic changes are almost entirely catalysed by biological systems in the microbe.

Alteration of these systems, so that the frequency of

unwanted genetic changes is decreased, could greatly extend the period of operation that is possible

before having to shut down and restart the fermentation. So far, this is a possibility only in microbes

that have a highly developed genetics. It may be that

strain stabilization of this sort would not be possible

in other microbes until their genetics are highly developed.

It is also possible to design strategies using current

strain development techniques that might lead to

genetically stable strains, but these are unproven.

● Cell separations. —Many fermentation schemes

incorporate cell recycle to boost productivity. This

requires that cells be separated from effluent broth.

Others need to separate cells from other residue as a

byproduct. In addition, some of the low-energy alternatives to distillation, such as adsorption, could require separation of the cells from the broth prior to

ethanol recovery.

In these cases, microbes that can be made to flocculate and redisperse, or that can be made to reversibly change their morphology would allow cheap

gravity separations (settling or flotation).

● Operating conditions.—An increase in the

temperature an organism will tolerate is advantageous for heat removal and in situ ethanol removal

schemes. The feasibility of accomplishing this is

uncertain.

The extreme of productivity improvement via cell

recycle is an immobilized cell reactor, It is conceivable that cells could be made less prone to

degradation under the conditions of immobilization,

by modifying sensitive components and degradation

systems, and by adding protective systems. This is

not at all a near-term possibility.

● Range and efficiency of substrate utilization.-A

single-step conversion of a substrate to ethanol is

highly desirable. This often requires that the ethanol

fermenting organism possess a degradation capability it does not have.

As an example, consider ligno-cellulose. It consists

of hexosans, pentosans, and lignin, All of these components should be used. Assume that one cellulaseproducing candidate does not use pentoses, while a

related noncellulase producing organism does, this is

exactly the situation with clostridia. If the second

organism can be given the cellulase genes of the first,

a microbe better-suited to direct conversion could be

created. The pace at which such a manipulation

could be developed cannot be predicted with confidence, although this is not necessarily a long-term

prospect.

Another obvious area that merits attention is the

enhancement of cellulase activity. Classical genetic

manipulations, employing mutation and selection or

screening, should result in micro-organisms better

equipped to degrade cellulose. E.g, it should be possible to isolate strains that are deregulated in cellulase

production (hyperproducers) as well as those in

which the cellulase is not subject to product inhibition. In addition, it is tempting to think about the

possibilities of amplifying cellulase genes by means

of DNA technology and cloning. However, this latter

approach must await further understanding of the

biochemistry and genetics of the cellulase system as

well as the development of the appropriate genetic

systems in cellulolytic micro-organisms,

Utilization offermentation byproducts

Presently for each gallon of ethanol produced, approximately 14 liters of stillage is formed. 17 If ethanol

is mixed with gasoline to make gasohol (10 percent

ethanol), the total stillage produced annually in the

United States would be in the billions of liters. Surely

a problem of this magnitude deserves serious attention. The utilization of stillage or fermentation byproducts could be greatly improved by genetic

means in several ways, In actuality, only a rational

long-range genetic approach can increase the value

of such a fermentation byproduct. Value can be increased in two main ways. The first is to increase the

nutritive value of the fermentation byproduct followed by developing economical processing technoli~w. E. Tyner, “The Potentjal of Obtaining Energy From Agriculture, ~Wposium on Biotechnology The Energy Production and Conservation, Gatlinberg, Term., 1979.
Appendix l-D—The Impact of Genetics on Ethanol—A Case Study ● 303

ogies that stabilize and preserve nutritive value. The

second approach is to increase the functionality of

the byproducts so that more useful products can be

developed.

For this one can envisage clever and novel ways to

utilize mutants to increase the value in a manner

similar to those described. 18 l9 20 Ethanol production

is not compatible with producing a valuable byproduct. E.g., a filamentous yeast may be useful for direct

texturization or fortification of an animal food but

production of ethanol may not be suitable with such

an organism. A possible solution to this type of conflict involves the development and engineering of

two-stage fermentation processes. In the first stage,

ethanol producing organisms are propagated under

optimal economic conditions for ethanol production.

After the production phase is over, the organisms

are then transferred to a second-stage reactor,

where desirable phenotypic properties are then expressed. Signals for expression of phenotypic properties can be extrinsic environmental parameters,

such as temperature, or levels of oxygen or carbon

dioxide, or intrinsic parameters, such as specific

nutrient requirements.

Thus the large-scale utilization of fermentation

byproducts as feed or other materials will then

become more valuable when genetic engineering can

decrease processing costs and increase product

laA. J, Sinskey, J. Boudrant, C. Lee, J. DeAngelo, Y. Miyasaka, (;. Rha, and S.

R. Tannenbaum, “Applications of Temperature-Sensitive Mutants for SingleCell Protein Production,” in Proceedings of U.S./U.S.S.R. Conference on Mechanisms and Kinetics of Uptake and Utilization of substrates in Processes for

the Production of Substances by Microbiological Means, Moscow-Pushchino,

p. 362, June 4-11, 1977. PB. 283-330-T.

19J, Boudrant, J. DeAngelo, A. J. Sinskey, and S. R. Tannenbaum, “process

Characteristics of Cell Lysis Mutants of Saccharomyces cerviciae,” Biotech.

Bioeng. 21:659, 1979.

ZOY, Mivasaka, A. J. Sinskey, J. DeAngelo, and C. Rha, “Characterization of

a Morphological Mutant of saccharomyces cervisiae for Single-Cell Protein

Production,” ./. Food Science 45:558 ;563, 1980.

quality. Most of these types of studies remain to be

done. However, the potential for innovative applications is great, but such applications may not result

because of the current lack of any Government agency that has a sound program for funding biotechnology research.

Recommendations and areas in which

applied genetics should have an impact

There has been little published research done in

the United States on the genetic improvement of

ethanol production processes with bacteria such as

Zymomonas and clostridia, and only limited studies

with yeast. In light of previous discussion, the following points have been identified as being the most important and relevant in the application of genetics

for improving ethanol-producing processes:

●

●

●

●

●

●

●

●

●

improvements on ethanol yield;

increased ethanol tolerance to achieve higher

final ethanol concentrations in the fermentation

broth;

increased rates of ethanol production;

elimination of unwanted products of anaerobic

catabolism, that is, direction of catabolism

towards ethanol;

enhanced cellulolytic and/or saccharolytic capabilities to improve rates of conversion of

cellulose and/or starch to fermentable sugars;

incorporation of pentose catabolic capabilities

into ethanol producers;

development of strains capable of hydrolyzing

cellulose and starch as well as of producing

ethanol from pentoses and hexoses;

improved temperature stability of micro-organisms and/or their enzymes; and

improved harvesting properties of cellular biomass produced during fermentation.
Appendix II-A

A Case Study of Wheat

Wheat is a major food staple in the diet of a large

percentage of the world’s population. Wheat grain in

the United States is used almost exclusively for

human consumption, although temporary localized

oversupply may result in some wheat feeding to livestock.

Attempts to improve wheat plant populations by

selection began several thousand years ago. The desirable attributes selected included the ability to

withstand severe environmental stresses such as

heat, cold, and drought and the stability of the seed

head (which tends to disarticulate in wild forms).

Wheat seeds moved from country to country

along with explorers and colonists. New varieties

played major roles in the establishment of many

productive wheat cultures—e.g., the Mennonite settlers introduced hard red winter (Turkey Red) wheat

into the Kansas area from Russia in the late 19th

century. And two private breeders—E. G. Clark of

Sedgenick, Kans., and Danne of Elreno, Okla.–developed varieties that set new levels of productivity

and straw strength in hard winter wheats which

were sought by millers for their excellent flour

recovery.

Breeding programs expanded during the first half

of the 20th century. At first, the U.S. Department of

Agriculture (USDA) played a lead role; hut the

emergence of the Land Grant System and the establishment of the State experiment station concept

prompted individual States to launch breeding programs designed to address the particular production

problems faced by farmers within their respective

boundaries.

As the State experiment stations began to assume

more responsibility, USDA programs and personnel

began to concentrate in central locations to assemble

the optimal number of personnel for the greatest interaction and productive output. If the present trend

continues, there will be virtually no USDA scientists

engaged in actual wheat breeding. Instead they will

have assumed the roles of basic researchers and regional coordinators supplying information to the

public and private breeders.

Disease and insect resistance have been the primary breeding goals of many programs. The dramatic losses associated with severe pest problems have

focused the attention of producers, researchers, and

legislators on these areas of need. Other traditional

breeding objectives have included improved use

properties, tolerance to environmental stresses such

as cold, wheat, wind dessication, and excessive moisture, and inherent yield capacity in the absence of

significant production limitations.

The quality of wheat’s end products has been

improved significantly through breeding. Varieties

have been tailored to meet the demands of various

industries. The bread bakeries needed a higher protein and more gluten strength to make a lighter,

larger loaf, while the cookie producer needed a lowprotein flour with desirable dough-spreading properties.

Wheat productivity and management

The pattern of wheat productivity (yield per acre)

in developed countries is remarkably similar. When

yields are plotted over the centuries, there is a long

period of barely perceptible increases in yield, from

the time of first records of production to the end of

the first third of this century (the period of 1925-35).

Since around 1935, yield has increased sharply. Recent data suggest that yield increases may be leveling

off. Why increases have been so substantial after

generations of little success, is a complex question involving genetic resources, economic development,

social interaction, and adoption of mechanical and

biological innovations.

Until recently, the U.S. commercial seed companies, with one or two exceptions, have not been interested in wheat breeding programs as a profitmaking venture. Since wheat has a perfect flower and

can fertilize itself, the farmer can purchase seed of a

new variety and reproduce it from generation to

generation. However, the discovery of cytoplasmic

male sterility and nuclear restorer genes has stimulated industry interest in the possibility of developing hybrid wheat. The farmer would purchase the

hybrid seed each year; the inbred lines used to make

the hybrid would be the exclusive property of the

originating company. Although progress has been

good, problems exist with the sterility and restorer

systems, the ability to produce adequate amounts of

hybrid seed, and the identification of economic levels

of hybrid vigor. The next 5 years should reveal the

potential for success in hybrid wheat.

Several milestones of progress have been set in

wheat. Yield has risen dramatically. Genetic protection against pests and other hazards has been a major contributor to increased yields. In addition, recent advances using semidwarf genes have been as304
Appendix II-A—A Case Study of Wheat . 305

sociated with significant yield improvement. The

shorter, stiffer stems of the semidwarf plants allow

maximization of resources without yield reductions.

Improvement in the inherent yield components of

stems per unit area, kernels per stem, and kernel

weight has also contributed extensively to yield improvement.

The use of applied genetics in wheat improvement

occurs in close harmony with total wheat management systems. The farmer must integrate a huge assortment of alternatives in each decision—e.g., an individual producer may be deciding on a nitrogen

program. If the farm is irrigated, the producer

selects nitrogen amounts and application timing

based on soil tests, intended crop and variety, the

end use of that crop, and watering schedules. [f the

farm is rainfed, the producer takes into account soil

tests, crop considerations, and rainfall probabilities.

In both cases product prices at the time of sale

must be predicted since they govern potential gross

return, which in turn affects the costs of maintaining

a profit margin. Genetic interaction in this system is

intricate. The farmer must first select the variety

most likely to produce at the maximum economic

level. For irrigated land, it may be a short highyielding semidwarf either for the cookie trade or the

export market. The farmer knows that part of the

value of his product is dependent on low protein.

However, inappropriately high levels of nitrogen,

which greatly improve yield, will also raise the protein of the crop beyond acceptable levels. If the export market is strong and the total U.S. supply reduced, the higher protein may be of little economic

consequence.

In the case of the dryland farmer, the variety

selected may be taller with lower yield potential but

with much better levels of adaptation and tolerance

to adverse environments. It may be designed for the

bread industry or the export market. Part of the

value is related to high-protein content. Since moisture conservation and use is critical, nitrogen applications and amounts must be selected so that the

plants do not waste their moisture reserve. However,

nitrogen applied too late may not receive enough

rain to penetrate the soil and become available to the

plants. If the plants “burn up” because of unwise

water use early in the season, the seeds will be high

in protein but low in yield. If inadequate nitrogen is

available, the crop will generally be low in protein.

The abbreviated protein story is but one of many

examples of farm management interaction with applied genetics in wheat production. Recent changes

in energy price and availability, environmental restraints, marketing structures, and technology development are producing a new array of complex problems.

Genetic vulnerability in wheat

Genetic vulnerability is defined as a high degree of

genetic uniformity in a crop grown over a wide acreage. Wheat, which is produced on about 62 million

acres annually in the United States, has a relatively

high level of uniformity and genetic vulnuerability.

In 1974,102 hard red winter wheat varieties were

grown on 36.6 million acres, with four varieties occupying 40 percent of the acreage. Hard red spring

wheat varieties totaled 80 percent on 14.7 million

acres, with three varieties occupying 52 percent of

the acreage. Similar situations occurred with other

classes of wheat. Plant pests, including diseases and

insects, have periodically caused moderate to servere

wheat crop losses in years favorable to the development of strains capable of attacking current forms of

resistance.

Incorporating genetic resistance to pests has traditionally been the responsibility of public breeders.

Wheat is a self-fertilized plant that can be faithfully

reproduced from generation to generation. Private

industry has been reluctant to invest R&D money in

improvements since the farmer, following the initial

seed purchase, can reproduce the crop without returning to the seed company. Thus, public breeders

have been the main source of new varieties and have

had the responsibility of delivering genetic improvements to the producer. Wheat breeding programs are generally designed to respond to State production needs. Goals and objectives are established

by technical advisory groups that include breeders

and scientists, growers, use industry representatives, and extension workers.

Genetic variability is available to the breeder from

naturally occurring sources and artificially induced

mutations. Naturally occurring variability has been

collected from native plant populations throughout

the world and is maintained in the World Wheat C ollection by the Science and Education Administration

of USDA located in Beltsville, Md. Currently, about

37,000 accessions are contained in the collection.

Breeders use the collection as a reservoir from which

to draw exotic genes needed to improve the value of

their breeding programs. In addition to variability

within wheat varieties, the breeders can use special

genetic techniques to draw valuable genes from related species such as rye and various forage grasses.

This approach, while time-consuming and costly, has

been used in a number of variety development programs. Mutations induced by artificial means have
306 ● lmpacts of Applied Genetics-Micro-Organisms, Plants, and Animals

not been used extensively by the breeders, since ly evaluated, characterized, and documented, forcdesired mutations without detrimental effects are ing breeders to spend time and resources carrying

very difficult to obtain. Enough natural genetic out their own evaluation work. The committee has

variability seems to exist to satisfy needs in the proposed a standard set of descriptors for all accesforeseeable future. sions in the collection, as well as an information

The National Wheat Improvement Committee has management system to efficiently bring the informastated that the World Wheat Collection is inadequate- tion to the breeders.
Appendix II-B

Genetics and the Forest Products

Industry Case Study

The Weyerhaeuser Co.

The Weyerhaeuser CO., which has its main headquarters in Centralia, Wash., is the largest forest

products company in the United States. In 1970,

Weyerhaeuser initiated a program to research the

mass propagation of Douglas fir trees by tissue

culture. Douglas firs are the main species in many of

the Nation’s forests, over $3.1 billion (or about 8.5

billion board feet) worth were harvested in 1979.

While they are normally propagated by seed in the

field, the classical development of improved seed

does not adequately satisfy the criteria of the rapid

availability of trees of superior quality.

Specially selected clones have the potential to double the productivity of forestlands; each year that

unimproved trees are planted is another year of

“suboptimum” harvests 40 years from now. With the

steadily increasing demand for forest products,

planting substantially improved trees as soon as possible is of great economic importance.

Weyerhaeuser’s tissue culture research began in

1974 with a project at the Institute of Paper Chemistry to produce Douglas firs. The project was expanded with a contract for additional research at the

Oregon Graduate Center. Although the intention was

to propagate select strains of mature trees, the main

focus of the program, in 1974 to 1978, was to develop a basic, consistent system for propagation. From

1978 to the present, Weyerhaeuser has been conducting most of its applied research into Douglas firs

at its own research facilities in Centralia, Wash. Basic

research is still being funded at the Institute of Paper

Chemistry, which services the entire forest industry.

While specific figures for the tissue culture systems

research have not been made available, the annual

research and development budget at Weyerhaeuser

specifically for biological work with forest species is

on the order of $7 million to $8 million. ’

The project in mass propagation of Douglas fir by

tissue culture was initiated to establish a reliable,

economic means for mass production of superior

trees. The cloning of these trees could bring higher

1 llRX M(’(kdlough, The \V(>}ffll-llat?lls(>l’ (k)., persomd (sonln~lllli(’ali(]il [Ma.v

1 !)80) with the I%inl Resourc[ls Institute in the working report, Commercia/

[ Isf.s Of planf I’issup (:1~/fure and Rjfenfial [mparf of Genetic Engineering on

l.’orf-.sl~y, prfy)iirfd lilldf?l’ (’olltl’il(’t 1[) () ”l’,\, 1980

yields and shorter harvest cycles, as well as rapid

production of tree stands for seed production.

The immediate results of 10 years of research are

not overly impressive at first glance. To date, 3,000

tissue-cultured Douglas firs have been planted for

comparison analysis and research of handling techniques, transfer procedures, etc.

The cost effectiveness of a tissue culture program

is determined by several factors, of which labor intensity varies the most. The more streamlined the

system can be made, the fewer labor-requiring steps

that are needed—the less direct costs will be incurred. Ideally, cells would be cultured in sterile conditions and then planted for the direct embryogenesis of plantlets that are ready for the field. Steps that

involve cutting shoots and rooting them on another

media or repeated subculturing procedures are costly and cumbersome. The major problem affecting

cost so far is the difficulty of achieving high volume

plant regeneration from the tissue cultures. Efficient

systems with more successful regeneration will reduce the labor and materials involved in culturing

and result ultimately in a lower cost per plant.

In addition to problems of cost, Weyerhaeuser has

run into the classic difficulty with woody species—

the inability to obtain required results from plants

more than 1 year old. In addition, the risk of induced

genetic variability increases with every subculture of

the tissues. The triggering techniques for effective

manipulation of mature versus embryonic and immature tree tissues are not well understood, and unlocking the Douglas fir system may well provide insight into some basic physiological questions.

Some commercial companies do not want to get

deeply involved in basic research because it is extremely expensive and time-consuming. However, it

has been up to the major forestry companies, such as

Weyerhaeuser, to independently fund essentially

basic research into the biological triggers for organogenesis and embryogenesis of Douglas fir.

By comparison, no other plant has been as intensely researched for mass propagation purposes and

proved so unyielding. Among other things, this indicates that questions of basic plant cell physiology

will have to be addressed before major breakthroughs can be expected. The goals of the Weyerhaeuser program are exacting and demand the refinement of present techniques into a precise in307
308 ● Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

dustrial science. While it may seem that the investment has been disproportional to the returns at this

point, it must be remembered that they are the forerunners of a new technology, both in terms of working with mature tree tissues of an especially intricate

species and in terms of imposing stringent industrial

standards on a mass biological production system.

Simpson Timber Co.

The Simpson Timber Co., whose central headquarters are in Seattle, Wash., is a large producer of redwood and other forest products, and has been involved over the past 5 years in a program to develop

a mass production system for the coast redwoods

through tissue culture. Approximately $250,000 has

been invested in research performed at the University of” California, Irvine, by Dr. Ernest Ball, a recognized authority in the field of tissue-cultured redwoods. z

Coastal redwoods are normally a field-seeded crop

and have a production cycle of around 50 years. The

major reason for consideration of tissue culture over

seed is the greater speed with which superior trees

might be developed through tissue culture as compared to using seed stock. Simpson Timber Co.,

which has been involved in a controlled breeding

program along conventional lines as well, and is approaching the creation of homozygous strains. Since

a sequoia seedling does not reach sexual maturity

before it is 15 to 20 years old, and since about 1 0

generations are normally required to produce a true

homozygous strain,3 the classical process is time-con.

suming and contains no guarantees that the end

products will be better than the clones selected

through tissue culture.

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Elite trees are selected from wild stands for

straightness of trunks, height, specific gravity of

wood, and proper branch drop (branches that drop

without tearing the stem). There are no major pests

in redwoods, so pest and disease resistance have not

been a concern. Two methods of selection are used.

Clones of special trees are produced by rooting the

uppermost branches of the tree, a process that takes

up to 1 year. Although the rooting percentage may

be as low as 10 percent, this method has the advantage of producing mature cloned plants that can continue to produce flowers and seed. Simpson is using

roughly 200 elite trees for these clones.

Mite trees can also provide clones through tissue

cultures of their needles, a process that is less timeconsuming but which produces seed very slowly

because of the time involved in maturation. Simpson

Timber Co. has planted out 2,500 tissue cultured redwoods for field comparisons with seedling material.

The results so far have been encouraging, but it may

take another 10 to 15 years before definite conclusions can be drawn. The major factors being analyzed are field growth rates and outplanting survival

percentages. Clones of elite varieties will also have to

be compared to the parent trees for the traits originally selected for, such as wood quality. Since the

operation] cost of tissue-cultured plantlets is about

twice that of seedlings, the quality of tissue cultured

plants must be markedly superior if the program is

to be cost effective,

Dr. Ball is confident that the tissue culture system

which has been developed for the rapid multiplication of elite redwood trees is ready for: implementation at a commercial production facility.4 Simpson

Timber (:0. is planning the construction of a tissue

culture lab at their California headquarters within

the next 2 years, The pilot plant is expected to cost

$250,000 and produce upwards of 200,000 plantlets

in its first year of production. As mass production

techniques are perfected, the company plans to expand the facility to a production capacity of over 1

million plantlets per years

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Appendix II-C

Animal Fertilization Technologies

Sperm storage

DEFINITION

The freezing of semen to – 196° C, storage for an

indefinite time, followed by thawing and successful

insemination.

STATE OF THE ART

Conception rates at first insemination with frozen

sperm average between 30 to 65 percent for most

species. This technology is not a key to the success of

artificial insemination (AI), but because of the convenience it is now an essential ingredient. Current

operational procedures are adequate for the dairy industry.

ADVANTAGES

1. Greater use of selected bulls as AI studs.

2. Elimination of the need to maintain expensive and

dangerous bulls on dairy farms.

3. Sperm can be tested for disease and treated for

venereally transmitted diseases.

4. Ease of transport and therefore of increasing potential offspring.

FUTURE

Little change is anticipated in semen processing.

Freeze-dried semen is unlikely to be successful

enough to use. Sperm banking can be expected to increase, especially on AI studs. Banking provides

cheap storage while bulls (slaughtered) are being

progeny tested, and insurance against loss of bulls

through natural causes. For preservation of semen

from bulls of less populous breeds, banking can be

completed in about ayear, after

be slaughtered.

Artificial insemination

DEFINITION

which the-bull can

Manual placing of sperm into the uterus.

STATE OF THE ART

Highly developed for most species. Representative

use rates in the United States are: dairy cattle, 60 percent; beef cattle, 5 percent; turkeys, 100 percent.

The major limitation to the use of AI is the low national average conception rate at first service,

around 50 percent. The success or failure of AI is

determined by a multiplicity of factors including

estrus detection, quality of semen, timing of insemination, and semen handling.

DISADVANTAGES

1. With increased herd size, estrus detection has

become a major problem.

2. Inexperienced dairymen are buying semen and inseminating their own cows, resulting in lowered

fertility and no feedback on semen fertility.

ADVANTAGES

1. Widespread use of genetically superior sires.

2. Services of proven sires at a lower cost.

3. Elimination of cost and danger of keeping bulls on

the farm.

4. Control of certain diseases.

S. Use of other breeding techniques including crossbreeding.

6. Continued use of valuable sire after his death.

FUTURE

Greater use of AI in beef cattle will depend on the

availability of successful and inexpensive estrus synchronization technology, on relaxed restrictions of

the various breed associations, and on accurate progeny records,

Estrus synchronization

DEFINITION

Estrus (“heat”), is the period during which the

female will allow the male to mate her. This sexual

behavior is subtle and varies widely among individuals. Thus the synchronization of estrus in a herd,

using various drug treatments, greatly enhances AI

and other reproduction programs.

STATE OF THE ART

Effective methods for synchronization of estrus

periods for large numbers of animals have been

available for more than two decades, and several approaches are now available which result in normal

fertility. Several schemes involve use of prostaglandin F2 (PGF2

) for the cow and ewe. However, FDA

approves usage only for controlled breeding in beef

cows and heifers, nonlactating dairy heifers, and in

mares.

ADVANTAGES

1. Time a heifer’s entry into a milking stream.

2. Increase productivity by breeding heifers earlier

in life.

309

76-565 0 - 81 - 21
310 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

3.

4.

to

in

Ability to breed large numbers of cattle over a

shorter calving interval.

Increase use of AI, especially in beef cattle, sheep,

and swine.

FUTURE

Estrus cycle regulation should allow selected sires

be more widely used to improve important traits

beef cattle. It should also gain widespread and

rapid acceptance among dairymen as well. -

Superovulation

DEFINITION

Superovulation is the hormonal stimulation of

multiple ovarian follicles resulting in release from

the ovary of a larger number of oocytes (ova) than

normal.

STATE OF THE ART

Superovulation with implantation into surrogate

mothers increases the number of offspring, usually

from highly selected dams. Adequate procedures are

presently available for superovulation of laboratory

and domestic animal species, except the horse. The

drugs used to induce superovulation are the gonadotropins, pregnant mare’s serum gonadotropin

(PMSG) and follicle stimulating hormone (FSH), in

some instances followed by other treatments to stimulate ovum maturation and ovulation. Superovulated

ova result in normal offspring with the same success

rates as achieved with normally ovulated ova.

DISADVANTAGES

1. Greatest drawback is that degree of success cannot be predicted for an individual animal.

2. Batches of hormones for ovulation treatment vary

widely in quality.

3. PMSG is scarce, and has been declared a drug by

the Food and Drug Administration (FDA). Thus,

most use of PMSG is now illegal.

4. There is insufficient data to judge the effect of

repeated superovulation.

FUTURE

Methods for superovulation will improve consistency of results. Additional understanding of basic

physiological mechanisms will facilitate such efforts.

New work in superovulatory technology involves active immunization against adrostenedione (a hormone involved in regulation of follicular development). This treatment prevents atresia and reliably

increases the frequency of multiple ovulations. The

technology has definite commercial potential for cattle husbandry and limited potential for sheep husbandry, and much current effort is directed towards

developing and testing a commercial procedure.

Embryo recovery

DEFINITION

The collection of the fertilized ova from the

oviducts or uteri. Collection of embryos is a

necessary step for embryo transfer or storage, and

for many experiments in reproductive biology. Both

surgical and nonsurgical methods are used.

STATE OF THE ART

Surgical. -Methods are available for recovering

40 to 80 percent of ovulations from cattle, sheep,

goats, swine, and horses. The development of adhesions and scar tissue following surgery limits these

techniques. Surgical recovery is the only method for

sheep, goats, and pigs. It is presently practiced

almost exclusively when a suspected pathology of the

oviducts renders an individual subfertile, or when

embryos must be recovered before the individual

reaches puberty.

Nonsurgical.- Non-surgical embryo recovery

techniques are preferred for the cow and horse.

Fifty to eighty percent of cow ovulations can be

recovered, and 40 to 90 percent of the operations on

horses to recover the single ovulation are successful.

ADVANTAGES

1. Nonsurgical embryo transfer can be performed an

unlimited number of times.

2. Requirements for equipment, personnel, and time

are low in nonsurgical recovery. This is especially

important in milk cattle: since the nonsurgical procedure is performed on the farm, milk production

is not interrupted.

3. A single embryo can be obtained between superovulation treatments.

4. Embryos can be obtained from a young heifer

before it reaches puberty.

5. The technology is especially important for research, e.g., in efforts to produce identical twins,

embryo biopsies for sex determination, etc.

FUTURE

Methods of collecting embryos have not changed

appreciably since about 1976, nor are significant advances predicted for the future.

Embryo transfer

DEFINITION

Implantation of an embryo into the oviduct or

uterus.
Appendix II-C—Anim81 Fertilization Technologies ● 311

STATE OF THE ART

Surgical.—Pregnancy rates of 50 to 75 percent

are achievable in cows, sheep, goats, pigs, and

horses. Surgical transfer is the only practical method

in sheep, goats, and pigs, and is the predominant

method for cows and horses. A number of factors

determine the success of surgical transfer: age and

quality of embryos, site of transfer, degree of synchrony between estrous cycles of the donor and recipients, number of embryos transferred, in vitro

culture conditions, skill of personnel, and management techniques. The 50- to 60-percent success rate

in cattle compares with AI success rates at first service. (Pregnancy rates should not be confused with

survival rates, which may be much lower.)

Nonsurgical. —This method is an adaptation of

AI. Reported success rates are much lower than

those with surgical transfer. Nonsurgical transfer is

not used in sheep, goats, or pigs.

1.

2.

3.

4.

5.

6.

7.

1.

2.

ADVANTAGES

Obtaining offspring from females unable to support pregnancy,

Obtaining more offspring from valuable females.

With a homozygous donor, undesirable recessive

traits among animals used for AI can be rapidly

detected.

Introducing new genes into specific pathogen-free

swine herds.

Coupled with short- or long-term embryo storage,

transportation of animals as embryos.

Increasing the population base of rare or endangered breeds of animals by use of closely related

breeds for recipients.

Separation of embryonic and maternal influences

in research.

DISADVANTAGES

Personnel requirements in surgical transfer account for a large share of high costs and thus limit

applicability in animal agriculture.

Provision of suitable recipients is the greatest

single cost in embryo transfer.

FUTURE

Surgical transfers will remain the method of

choice for sheep, goats, and pigs in the foreseeable

future. For cows and horses, however, nonsurgical

methods will be increasingly used rather than surgical techniques (and this will be apparent) within

the next year or two. It is likely that half of the commercial transfer pregnancies in cattle in North America in 1980 will be done nonsurgically, even if success rates are only 60 to 80 percent of those obtainable with surgical transfer. Among future applications, a role for embryo transfer can be predicted

in progeny testing of females, obtaining twins in beef

cows, obtaining progeny from prepubertal females,

and in combination with in vitro fertilization and a

variety of manipulative treatments (e.g., production

of identical twins, selfing, genetic engineering, etc.)

Embryo storage

DEFINITION

Maintenance of embryos for several hours or days

(short-term) or for an indefinite length of time (freezing).

STATE OF THE ART

Short-term.-The requirement for embryos

from farm animal species has not been defined,

although adequate culture systems for the short interval between recovery and transfer have been

developed by trial and error. Whereas the important

parameters of culture systems have been identified

(e.g., temperature, pH, etc.), optimal conditions have

not been determined. Cow embryos may be stored

for three days in the ligated oviduct of the rabbit.

Long-term (freezing).-No completely adequate

protocol exists for freezing embryos of farm species.

One-third to two-thirds of embryos are killed using

present methods. Pregnancy rates of 32 to 50 percent for cattle, sheep, and goats have been reported

after freezing. No successful freezing of swine or

horse embryos followed by development to term has

been reported. Despite disadvantages (one-half of

embryos are often killed) advantages are such that in

some’ situations embryo freezing, and embryo selling, are already profitable.

ADVANTAGES

1. Amplification of advantages of embryo transfer.

2. Elimination of requirements for large recipient

herds when embryo transfer is being used.

3. Reduction of costs in animal transport.

4. Control of genetic drift in animals over prolonged

time intervals.

FUTURE

Anticipated development of embryo culture technology would be of significance in efforts toward in

vitro maturation of gametes, in vitro fertilization, sex

determination, cloning, and genetic engineering, all

of which involve prolonged manipulation of gametes

and embryos outside of the reproductive tract.

As freezing rates improve, nearly all embryos recovered from cattle in North America will be frozen.

Probably as many as half of the embryos will be

deep-frozen for 2 to 3 years. It is unlikely that success rates will ever approach 90 percent of those
312 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

without freezing. However, 70- to 80-percent success

rates may be attainable within several years. It appears that embryos can be stored indefinitely with

little deterioration.

Sex selection

DEFINITION

Tests to determine the sex of the unborn or determination of sex at fertilization by separating x- bearing from y-bearing sperm.

STATE OF THE ART

Sexing of embryos.—Through karyotyping

nearly two-thirds of embryos can be sexed. Techniques using identification of the condensed X chromosomes are unreliable. A third method, identification of sex-specific gene products, is under development.

Sexing of sperm.—A 100-percent method has

not been achieved in any mammalian species; and no

standard protocol for farm species exists.

FUTURE

Before this technology can be applied commercially, it must be simple, fast, inexpensive, reliable, and

nonharmful for embryos. Such techniques could undoubtedly be developed. There would be numerous

medical and experimental applications.

There is much interest in research in this area

because of its use in understanding male fertility

with AI in humans, and in enhancing sperm survival

after frozen storage.

Twinning

DEFINITION

Artificial production of twins, either using embryo

transfer or hormone treatments,

STATE OF THE ART

Currently, embryo transfer is the most effective

method for inducing twin pregnancies in cattle,

resulting in pregnancy rates of between 67 to 91 percent, of which 27 to 75 percent deliver twins. Other

methods include transferring one embryo into a cow

which has been artificially inseminated, and hormonal induction of twinning, which is a modification

of superovulation. This latter method is not reliable.

ADVANTAGE

The advantage of twinning in nonlitter-bearing

species is the improved feed conversion ratio of producing the extra offspring.

DISADVANTAGE

The major disadvantage of twinning is intensive

management necessary for periparturient complications, unpredictable gestation periods, depressed lactation, etc.

FUTURE

Technical feasibility for twinning, in conjunction

with embryo transfer, management adjustments,

and selection for good recipients, can be predicted. A

reliable procedure for twinning in sheep can also be

expected. The technology would most likely be first

used in Europe and Japan, where there are shortages

of calves to fatten for beef.

In vitro fertilization

DEFINITION

The union of egg and sperm outside the reproductive tract. For some species, the technology includes

successful development of the embryo to gestation

and birth.

STATE OF THE ART

In vitro fertilization has been accomplished in

several laboratory animal species, including the rabbit, mouse, rat, hamster, and guinea pig and nine

other mammalian nonlaboratory species, including

man, cat, dog, pig, sheep, and cow. However, normal

development following in vitro fertilization and embryo transfer has only been accomplished in the rabbit, mouse, rat, and human. Consistent and repeatable success with in vitro fertilization in farm species

has not yet been accomplished.

None of the cases of reported success of in vitro

fertilization, embryo transfer, and normal development in man is well documented.

Most of the in vitro fertilization work to date has

concentrated on the development of a research tool

so that the physiological and biochemical events in

fertilization and early development could be better

understood. More practical application of in vitro

fertilization techniques would include:

1. a means for assessing the fertility of ovum

and/or sperm;

2. a means to overcome female infertility with embryo transfer into a recipient animal; and

3. when coupled with ovum and/or embryo storage and transfer, a means to facilitate combination of selected ova with selected sperm for production of individuals with predicted characteristics at an appropriate time.
Appendix /l-C—Animal Fertilization Technologies ● 313

FUTURE

Rapid progress in research is anticipated and

many of the potential applications of in vitro fertilization to animal breeding should become practical

within the next 10 to 20 years. With further development of in vitro fertilization methodology, along with

storage of unfertilized oocytes (gamete banking), fertilization of desired crosses should become possible.

In the more distant future, genetic engineering and

sperm sexing along with in vitro fertilization may

become possible.

Parthenogenesis

DEFINITION

The initiation of development in the absense of

sperm.

STATE OF THE ART

Parthenogenesis has not been satisfactorily demonstrated or described for mammalian species. The

best available information leads to the conclusion

that maintenance of parthenogenetic development to

produce normal offspring in mammals approximates

impossibility.

Cloning: production of identical twins

DEFINITION

The production, using a variety of methods, of

genetically identical individuals.

STATE OF THE ART

There are several ways to obtain genetically identical livestock. The natural way is identical twins,

although these are rare in species other than cattle

and primates. Both natural and laboratory methods

depend on the fact that the blastomeres of early embryos are totipotent (i.e., each cell can develop into a

complete individual if separated from the others.)

For practical purposes, highly inbred lines of some

mammals are already considered genetically identical; F1 crosses of these lines are also considered

genetically identical and do not suffer from the

depressive effect of inbreeding.

ADVANTAGE

An advantage of identical twins is the experimental control provided by one animal through which

two sets of environmental conditions can be compared for effects on certain end points, e.g., native v.

surrogate uterine environments for gestational development, nutrition on milk production, etc.

Cloning: nuclear transplantation

DEFINITION

The production of genetically identical mammals

by inserting the nucleus of one cell into another,

before or after destroying the original genetic complement. These occur by separation of embryos or

parts of embryos early in development but well after

fertilization has occurred.

STATE OF THE ART

Experimentalists have found in certain amphibia

that transplantation of a nucleus from a body cell of

an embryonic (tadpole) stage into a zygote following

destruction or removal of the normal nucleus can

lead to development of a sexually mature frog.

FUTURE

The ideal technique for making genetic copies of

any given outstanding adult mammal would involve

inserting somatic (body) cell nuclei into ova, which

may take years of work to perfect if indeed it is possible, There is some evidence that adult body cells are

irreversibly differentiated.

How identical will clones be? They can be expected to be fairly similar in appearance. They would

be less similar than identical twins, however, which

share ooplasm and uterine and neonatal environments. Furthermore, certain components are inherited exclusively from the mother, e.g., the mitochondrial genome and perhaps the genome of centrioles.

The random inactivation of one or the other of the X

chromosomes may also limit similarities. Other differences among clones would result from the prenatal environment: in litter-bearing species even

uterine position can affect offspring. In single-bearing species the maternal effect may be pronounced.

Environmental differences in later life may greatly

affect certain traits, even if those traits have a strong

genetic component.

Serious technical barriers must be overcome

before realistic speculation of possible advantages in

animal production can be foreseen.

cell fusion

DEFINITION

The fusion of two mature sex cells or the fertilization of one ovum with another. An analogous scheme

for the male would be accomplished by microsurgical removal of the female pronucleus and substitution of nuclei from two sperm. Combining sex cells

from the same animal is called “selfing.”
314 . Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

STATE OF THE ART

Combination of ova has led to early development

to the blastocyst stage in the mouse but no further

development following transfer has been reported.

Initial success in experimentation with manipulation

of pronuclei has been reported.

FUTURE

Cell fusion technology may someday prove useful

for getting genetic material from a somatic cell into a

fertilized l-cell embryo for the purpose of cloning. In

conjunction with tissue culture technology the technology would have a role in gene mapping of chromosomes for the cow and perhaps other species.

Combining ova of the same animal, selfing,

would rapidly result in pure genetic (inbred) lines for

use as breeding stocks. The technique would also

lead to rapid identification of undesirable recessive

traits which could be eliminated from the species.

Chimeras

DEFINITION

A chimera is an animal comprised of cell lines

from a variety of sources. They can be formed by

fusing two or more early embryos or by adding extra

cells to blastocysts.

STATE OF THE ART

Live chimeras between two species of mouse have

been produced. Such young have four parents instead of two; hexaparental chimeras have also been

produced.

FUTURE

Practical applications of chimera technology to

livestock are not obvious at this stage of development. The main objective of this research is to provide a genetic tool for better understanding of development, and maternal-fetal interactions.

Recombinant DNA

DEFINITION

The introduction of foreign DNA into the germplasm.

STATE OF THE ART

The mechanics of changing the DNA molecules of

farm animals directly have not yet been worked out.

The plasmid methods used in bacteria may not be applicable,

FUTURE

None of these techniques, no matter how great the

potential, will be of any use in animal breeding until

knowledge of genetics is greatly advanced. Before

one can alter genes, they must be identified,

Prior to exploitation of recombinant DNA technology in animal breeding, it is necessary to identify

gene loci on chromosomes, i.e., genetic mapping.

Work toward this goal has only recently been initiated and rapid progress cannot be anticipated. Multivariate genetic determinants of characteristics of

economic importance are anticipated to be the rule.
Appendix III-A

History of the

Recombinant DNA Debate

The history of the debate over the risks from

rDNA techniques and the Government’s response

may be divided into four phases. * Phase I covered

the period from the first awareness of risks to

human health from experiments involving recombinant DNA (rDNA) in the summer of 1971 to the end

of the Conference at the Asilomar Center in February 1975, which resulted in prototype guidelines

covering the research, Phase 11 covered the period

from Asilomar through the development by the National Institutes of Health (NIH) of the Guidelines of

June 1976. In this period, the public first became

significantly involved in the debate and most, if not

all, of the policy issues were clearly framed. Phase

111, from mid-1976 through mid-1978, involved congressional consideration of the issues in an atmosphere that went from almost imminent passage of

legislation to the cessation of such efforts. Phase IV

covers the postlegislative period, when NIH and its

organizational parent, the Department of Health,

Education, and Welfare (HEW) (now the Department

of Health and Human Services) undertook to develop

satisfactory voluntary standards in areas over which

they had no legal authority and to accommodate

growing pressure for public involvement, while

avoiding a full regulatory role.

Phase I began in the summer of 1971, when several scientists became concerned about the safety of

a proposed experiment to insert DNA from SV40

virus, a monkey tumor virus that also transforms human cells into tumor-like cells, into a type of bacteria

naturally found in the human intestine. After

months of discussion, the scientist who had proposed the experiment decided to defer it. Meanwhile,

as rDNA techniques became more refined, debates

about safety increased; at the June 1973 Gordon

Research Conference, safety issues were discussed.

The participants voted: to send a letter to the National Academy of Sciences (NAS) and the National In-

“For a detailed history through 1977, see footnote 1. For a history and a discussion of the broader issues, see footnotes 2 and 3.

‘J. Swazey, J. Sorenson, and C. Wong, ‘(Risks and Benefits,

Rights and Responsibilities: A Historv of the Recombinant DNA Research Controversy,” Southern California Law Review SI:1019,

September 1978.

‘C. Grobstein, A Double Image of the Double Helig (San Francisco: W. H. Freeman Co., 1979).

3D. Jackson, and S. Stich (eds. ), The Recombinant DNA Debate

(Englewood Cliffs, N. J.: Prentice-Hall, Inc., 1979).

stitute of Medicine> requesting the appointment of

committees to study potential hazards to laboratory

workers and the public; and by a narrow majority4

to arrange for the letter to be published in the widely

read journal, Science, to alert the broader scientific

community.5

NAS appointed a committee of prominent scientists involved in rDNA research. In July 1974, the

panel asked for a temporary worldwide moratorium

on certain types of experiments, and called for an international conference on potential biohazards of

the research through a letter published in Science

and its British counterpart, Nature.6 This letter also

requested the Director of NIH to consider establishing an advisory committee to develop an experimental program to evaluate potential hazards and

establish guidelines for experimenters.

In response, the Director of NIH, after authorization by the Secretary of HEW, established the Recombinant DNA Molecule Program Advisory Committee

(later renamed the Recombinant DNA Advisory Committee, RAC) on October 7, 1974, along the lines suggested by the NAS Committee. The Committee’s

charter described its purpose as:7

The goal of the Committee is to investigate the current state of knowledge and technology regarding

DNA recombinant, their survival in nature, and

transferability to other organisms; to recommend

programs of research to assess the possibility of

spread of specific DNA recombinant and the possible

hazards to public health and to the environment; and

to recommend guidelines on the basis of the research

results. This Committee is a technical committee, establishedto look at a specific problem. (Emphasis added.)

The international conference called for by the

NAS Committee letter was held at the Asilomar Conference Center, Pacific Grove, Calif., in February

1975. The organizing committee made it clear that its

purpose was to focus on scientific issues rather than

to become involved in considering ethical and moral

questions. However, in one session the few lawyers

4Swazey, et al., op. cit., p. 1,023.

‘Letter from Maxine Singer and Dieter Soil to the National

Academy of Sciences (NAS) and the National Institute of Medicine,

reprinted in Science, vol. 181, 1973, p. 1114.

%etter from Paul Berg, et al. to the editor, reprinted in Science,

VO]. 185, 1974, p. 303.

The charter of the Recombinant DNA Molecule Program Advisory Committee, Oct. 7, 1974.

315
316 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

invited confronted the scientists with some of these

questions.’ The conference report concluded that although a moratorium should continue on some experiments, most work involving rDNA could continue with appropriate safeguards in the form of

physical and biological containment,

In Phase II, the debate widened to encompass

broader social and ethical issues, such as the relationship between scientific freedom of inquiry and

the protection of society’s interests, in whatever

manner those were defined. Such issues led naturally to questions about who makes the decisions and

the role of the public in that process. Finally, decisionmaking mechanisms were developed. Issues

raised and actions taken during this phase in many

respects controlled the subsequent development of

the Federal response to the debate, and created

problems that continue to the present. At this stage,

participation in the debate went beyond the scientific community.

Questions of ethics and public’ policy had been

raised earlier, but they now received much wider attention. On April 22, 1975, Sen. Edward M. Kennedy,

Chairman of the Subcommittee on Health of the

Senate Committee on Labor and Public Welfare, held

a half-day hearing on science policy issues arising

from rDNA research. In May 1975, a 2-day conference on “Ethical and Scientific Issues Posed by

Human Uses of Molecular Genetics” was held under

the joint sponsorship of the New York Academy of

Sciences and the Institute of Society, Ethics, and the

Life Sciences. In addition to molecular biologists, participants included lawyers, sociologists, psychiatrists,

and philosophers.

The issue of public participation arose as decisionmaking mechanisms were developed. RAC was originally composed of 12 members from “the fields of

molecular biology, virology, genetics and microbiolOgy.”9 Critics first noted the need for more expertise

in the fields of epidemiology and infectious diseases,

since most molecular biologists were trained as

chemists. * RAC’s membership was increased to 16

and the range of expertise was widened to include

the fields of epidemiology, infectious diseases, and

the biology of enteric organisms, by amendment to

the charter on April 25, 1975.

Since some members were conducting the research in question, critics claimed that a conflict of

interests existed. They also noted that the Committee

%3wazey, et al., op. cit., p. 1,034.

Whe charter of the Recombinant DNA Molecule Program Advisory Committee, Oct. 7, 1974, op. cit.

● One of the members of the original RAC (Stanley Falkow) did

have substantial expertise with enteric organisms and E. coli in

particular.

advised the Director of NIH, an agency whose mission was to foster biomedical research, not to stop or

otherwise regulate it. These issues were brought out

in a petition to NIH signed by 48 biologists in August

1975. Criticizing a proposed draft of the guidelines as

setting substantially lower safety standards than

those accepted at Asilomar, the petition argued for

broader representation on RAC from other fields of

scientific expertise and from the public-at-large. RAC

itself had been sensitive to these limitations; in the

summer of 1975, an attempt was made to recruit

nonscientists.l0 One nonscientist was added in

January 1976, and another was added in August

1976.

In December 1975, RAC submitted revised draft

guidelines to the Director of NIH, Dr. Donald

Fredrickson, Although they were stricter than those

drafted at Asilomar, some criticized them as being

“tailored to fit particular experiments that are already on the drawing boards.”11 The consensus of

RAC, on the other hand, was that the guidelines were

excessively strict, but that it was necessary to be

overly cautious because of its limited expertise in

public health.l2 In any event, Dr. Fredrickson arranged for public hearings on the proposed guidelines at a 2-day meeting in February 1976 of the Advisory Committee to the Director, a diverse group of

scientists, physicians, lawyers, philosophers

) and

others. A similarly diverse group of scientists and

public interest advocates were invited to attend.

Some modifications to the Guidelines proposed by

Dr. Fredrickson as a result of that meeting were

adopted and others were rejected by RAC in April

1976. 13

The final major issue arising during this period

concerned NIH’s lack of authority to set conditions

on research funded by other Federal agencies or by

the private sector. In a June 2, 1976, meeting between Dr. Fredrickson and some 30 representatives

of industry, including pharmaceutical and chemical

companies, it became clear that some rDNA research

was being done; however, the representatives appeared hesitant to commit themselves to voluntary

compliance with the proposed guidelines.l4 The priIODr+ E]izabeth Kutter, a former RAC member, persona] cot’nmunication, Sept. 11, 1980.

1 IN, wade, ~,Re~ombinant DNA: NIH sets strict Rules to ~unch

New Technology,” Science, vol. 190, 1975, pp. 1175, 1179.

‘zKutter, op. cit.

‘sIbid.

ltsubcommittee on science, Research and Technology of the

House Committee on Science and Technology, Genetic Engineering

Human Genetics, and Cell Biology: DNA Recombinant Molecule Research (Supp. Report 11) 94th Cong,, Zd sess,, 1976, p, 51.
Appendix III-A—History of the Recombinant DNA Debate ● 317

mary reason was their concern over protection of

proprietary information.l5

Phase 11 culminated with the promulgation on

June 23, 1976, of the Guidelines for Research Involving Recombinant DNA Molecules (“1976 Guidelines”)

covering institutions and individuals receiving NIH

funds for this research.

Phase III was characterized by attempts to remedy

the limited applicability of the Guidelines. Soon after

their publication, Senators Kennedy and Javits sent a

letter to President Ford, calling his attention to the

Guidelines. They noted that any risk was not limited

to federally funded research, and urged him to

take necessary steps to implement the Guidelines

throughout the research community. In October

1976, the Secretary of HEW, with the approval of the

President, formed the Federal Interagency Advisory

Committee under the chairmanship of the Director

of NIH to determine the extent to which the Guidelines could be applied to all research and to recommend necessary executive or legislative actions to

ensure compliance.l6 In March 1977, the Committee

concluded that existing Federal law would not permit the regulation of all rDNA research in the United

States to the extent deemed necessary;17 it further

recommended new legislation, specifying the elements of that legislation, is

During 1977, several bills to deal with this and

other problems were introduced in Congress. They

addressed in different ways the issues of the extent

of regulatory coverage, the mechanisms for regulation, and Federal preemption of State and local regulation. The major bills were those of Rep. Paul

Rogers, H.R. 7897 (and its substitute, H.R. 11192) and

of Sen. Edward Kennedy, S. 1217. *

While hearings were being held, three developments occurred which, by the end of 1977, had

dissipated much of the impetus for legislation. The

first was the expanded role of RAC. On September

24, 1976, its charter had been amended once more to

provide for additional expertise in the areas of

botany, plant pathology, and tissue culture. Moreover, its membership was increased from 16 to 20 so

that four members would be “from other disciplines

or representatives of the general public. ” This was

the first official provision for public representation

‘

s

Ibid., pp. 52.

Iafnterim Repofl of the Federal Interagency Committee On flec0l7?-

binant DNA Research: Suggested Elements for Legislation, Mar. 15,

1977, pp. 3.4.

“[bid., pp. 9-10.

‘“Ibid., pp. 11-15.

● For a more complete discussion of the legislation, see footnote

19.

‘

g

.’’ Recombinant DNA Molecule Research, ” Congressional Research Service, issue brief No. IB 77024, update of Jan, 2, 1979.

although two nonscientists were already members.

The number of nonscientists remained the same

until December 1978.20 Also, RAC’s responsibilities

were defined in greater detail, including the responsibility for reviewing large-scale experiments. Nevertheless, RAC continued formally at least to be “a technical committee, established to look at a specific

problem.”

The second development was a growing belief

among scientists that the risks of the research were

less than originally feared. This was based on the following: 1) a letter from Roy Curtiss at the University

of Alabama to the Director of NIH, explaining risk

assessment experiments using Escherichia coli, from

which he concluded that the use of E. coli K-12 hostvectors posed no danger to humans; 2) the conclusions of a committee of experts in infectious diseases

assembled by NIH in June 1977 in Falmouth, Mass.,

that the alleged hazards of the research were unsubstantiated; and 3) a prepublication report on experiments showing that genetic recombination occurs naturally between lower and higher life forms,

and suggesting that the rDNA technique was not as

novel as presumed.

The third development affecting the legislation

was a concerted lobbying effort by scientists against

what they considered to be some of the overly

restrictive provisions of the bills, especially S.

1217.21 22 23 The efforts included wide circulation of

reports (including some in draft form) as soon as

available, which supported the conclusion that

the research was less hazardous than originally

supposed.

By the end of 1977, the legislation was in limbo.

This situation continued in early 1978, although

some hearings were held. On June 1, 1978, Senators

Kennedy, Javits, Nelson, Stevenson, Williams, and

Schweiker addressed a letter to HEW Secretary

Joseph Califano, which acknowledged the likelihood

that legislation would not pass and urged that deficiencies in the regulatory system be addressed

through executive action based on existing authority,

if that were to be the case.

During Phase IV, NIH and its parent organization,

HEW (now DHHS), have attempted to operate in the

regulatory vacuum left by the lack of legislation. In

response to the consensus that developed in 1977 on

~William Gartland, Director of the Office of Recombinant DNA

Activities, NIH, personal communication, June 19, 1980.

ZIB. Cu]]iton, “Recombinant DNA Bills Derailed: COngre55 Still

Trying to Pass Law,” Science, vol. 199, Jan. 20, 1978. pp. 274-277.

“D. Dickson, “Friends of DNA Fight Back,” Nature, vol. 272,

April 1978, pp. 664-665.

ZSR. ~win, ,, Recombinant DNA as a political pa~n,~’ ~e~ ~ien.

fist, VO]. 79, Sept. 7, 1978, pp. 672-674.

76-565 0 - 81 - 22
318 ● Impacts of Applied Genetics-Micro-Organisms, Plants, and Animals

the question of risk, RAC proposed revisions to the

Guidelines, which placed most experiments at a

lower containment level. They were published for

public comment in September 1977. * As with the

original Guidelines, public hearings were held in the

course of a 2-day meeting of the Advisory Committee

to the Director in December 1977, in which a diverse

group of individuals and organizations were permitted to comment. However, at this point, HEW took a

much more active role in a situation that had been

handled almost entirely by NIH.24

When RAC’s charter was renewed on June 30,

1978, Secretary Califano reserved the power to appoint its members instead of delegating it to the

Director of NIH as in the past.** And the new proposed Guidelines, published in the Federal Register

on July 28, 1978, were accompanied by an introductory statement by Secretary Califano announcing a

60 day public comment period to be followed by a

public hearing before a departmental panel chaired

by HEW General Counsel Peter Libassi. * * * The

Secretary was particularly interested in comments

on: new mechanisms to provide for future discretionary revision of the Guidelines; and the composition of the various advisory bodies, especially the

RAC and the local Institutional Biosafety Committees

(IBCS).25

The public hearing called for by Secretary Califano and held on September 15, 1978, was a significant event in the history of Federal actions on the

rDNA issue. Testimony was heard from representatives of industry, labor, the research community,

and public interest groups; more than 170 letters of

comment were received and subsequently reviewed.

As a result, the revised final Guidelines of December

22, 1978, were significantly rewritten to increase

public participation in the decisionmaking process:26

● Twenty percent of the members of the IBCs had

to represent the general public and could have

no connection with the institution.

● Most of the records of the IBCs had to be publicly available.

● Shortly thereafter, in October 1977, the Final Environmental

Impact Statement for the 1976 Guidelines was published.

Z4D. Fredrickson, “A History of the Recombinant DNA Guidelines in the United States,” Hecornbinmt DNA Technical Bulletin,

vol. 2, Ju]y 1979. pp. 87, 90.

● ● The statement providing for delegation of authority that accompanied the updated Charter was not signed by Califano. See

also, footnote 24.

* ● ● The other members of the HEW panel were Dr.

Fredrickson, Julius Richmond, who was the Assistant Secretary

for Health, and Henry Aaron, who was the Assistant Secretary for

Planning and Evaluation.

=43 F.R. 33042, July 28, 1978.

~estatement of Secretav Califano accompanying the revised

Guidelines, 43 F.R. 60080, Dec. 22, 1978.

● Major actions, such as decisions to except otherwise prohibited experiments on a case-by-case

basis or to change the Guidelines, could be made

only on the advice of RAC and after public and

Federal agency comment.

The increased public responsiveness of the IBC’s

was crucial, since the revised Guidelines placed major responsibility for compliance on them. This had

been proposed in the July version and had not been

changed by the hearings. * Califano also announced

he would appoint 14 new members to the RAC, including people knowledgeable in fields such as law,

public policy, ethics, the environment, and public

health. Z’* ● All of these changes were envisioned to

“provide the opportunity for those concerned to

raise any ethical issues posed by recombinant DNA

research” and to change the role of the RAC to “serve

as the principal advisory body to the Director of NIH

and the Secretary of HEW on recombinant DNA

Policy .’’28* * *

In addition to broadening public participation,

Califano attempted to deal with a major limitation of

the Federal response—the Guidelines did not cover

private research, He directed the Food and Drug Administration (FDA) to take steps to require that any

firm seeking approval of a product requiring the use

of rDNA techniques in its development or manufacture, demonstrate compliance with the Guidelines

for the work done on that product; an FDA notice of

its intention to propose such regulations accompanied the revised Guidelines in the Federal Register.

In addition, he requested the Environmental Protection Agency (EPA) to review its regulatory authority

in that area. He believed if both agencies could

regulate research on products within their jurisdiction, “virtually all recombinant DNA research in this

country would be brought under the requirements

of the revised guidelines. ’’29* In the meantime, the

● As part of the revision process, HEW held a meeting in October

1978 for IBC chairpersons in order to exchange information and

experiences gained under the 1976 Guidelines.

Z71bid.

● ● This was implemented by an amendment to the RAC Charter

on Dec. 28, 1978, which increased the membership to 25 and

changed the composition to the following categories: 1) at least

eight specialists in molecular biology or rDNA research; 2) at least

six specialists in other scientific fields; and 3) at least six persons

knowledgeable in law, public policy, the environment, and public

or occupational health. In addition, the Charter was amended to

grant nonvoting representation to representatives of various Federal agencies.

ZsIbid.

● * ● The Charter was never amended to change or delete the

final sentence of the “Purpose” section, which states, “This Corn.

mittee is a technical committee, established to look at a specific

problem.”

‘“Ibid., p. 60081.
Appendix I//-A—History of the Recombinant DNA Debate ● 319

revised Guidelines provided, for the first time, for

voluntary registration of projects with NIH, in which

the registrant would agree to abide only by the containment standards of the Guidelines.31

Other major changes were embodied in the new

Guidelines. Because of the consensus that the experiments posed lower risks than originally thought,

some types of experiments were exempted, while

containment levels were lowered for almost all

others. In order to provide greater flexibility, these

Guidelines permitted exceptions on a case-by-case

basis, and included procedures for their change on a

piecemeal basis without going through the whole internal process at HEW. For major changes, the procedure was: 1) publication of the proposed changes

in the Federal Register at least 30 days prior to a RAC

meeting; 2) RAC consideration of the proposed

changes; and 3) publication in the Federal Register of

the final decision of the Director, NIH. The standard

for all actions of the Director under the Guidelines

was “no significant risk to health or to the environmerit. ”32 Lastly, the new Guidelines delegated project

approval to the IBCs.

The problems posed by voluntary compliance and

commercialization have continued to be addressed

by NIH. In a second major revision to the Guidelines

on January 29, 1980, a section (Part VI ) was added to

specify procedures for voluntary compliance. * ● On

(cominuedfmm p. 318)

● Subsequently, Califano sent similar letters to the Secretaries of

Agriculture [February 1979) and Labor (July 1979) requesting

them to consider how their agencies’ authorities could be used to

require private sector rDNA research to comply with the

Guidelines. qo

aoMinutes of the Interagency Committee on Recombinant DNA

Research, p. 3, July 17, 1979, preprinted in Recombinant DNA Research, vol. 5, p. 132, et. seq.

alsec. IV.F.3, 1978 Guidelines.

3zSec. IV-E-l-b.

● “Several responses to the FDA notice had questioned the agen.

cy’s legal authority to regulate private rDNA research. Conse.

quently, Dr. Fredrickson and Dr. Donald Kennedy, then Commissioner of Food and Drugs, developed a draft supplement to the

Guidelines, specifying procedures for voluntary compliance by in.

dustry. It was published for comment on Aug. 3, 1979 (44 F.R.

45868) and incorporated as part of the proposed revised Guide.

lines of November 30, 1979. (44 F.R. 69210, 69247).

April 11, 1980, NIH published Physical Containment

Recommendations for Large Scale Uses of Organisms

Containing Recombinant DNA Molecules in the form

of Draft Part VII to the Guidelines.33 Besides setting

large scale containment levels, this document recommends that the institution: appoint a biological safety

officer with specified duties; and establish a worker

health surveillance program for work requiring a

high (P3) containment level. Finally, a more ad hoc requirement has been used since October 1979 for approvals of industrial requests for cultures up to 750

liters (1); the approvals were conditioned on NIH

designated observers being permitted by the companies to inspect their facilities.34 At least one inspection has taken place.

On November 21, 1980, NIH adopted the third major revision to the Guidelines.35 It contained these

significant changes: institutions sponsoring the

research are no longer required to register their

projects with NIH pursuant to an informational document called a Memorandum of Understanding (MUA)

whenever the containment levels are specified in the

Guidelines; and NIH will no longer review IBC decisions on experiments for which containment levels

are specified in the Guidelines.

On November 21, 1980, NIH also promulgated

revised application procedures for large-scale proposals. The application must include the following information: 1) the registration document submitted to

the local IBC; 2) the reason for wanting to exceed the

10-1 limit; 3) evidence that the rDNA to be used was

rigorously characterized and free of harmful sequences; and 4) specification of the large-scale containment level proposed to be used as defined in the

NIH Physical Containment Recommendations of

April 11, 1980.

In addition to adding part VI to the Guidelines, the most significant change in the January 1980 Guidelines was the addition of

sec. 111-0, which permitted most experiments using E. coli K-12

host-vector systems to be done at the lowest containment levels.

s345 F.R. 24%8, Apr. 11, 1980.

3444 F.R. 69251, NOV. 30, 1979.

3345 F.R. 77372, Nov. 21, 1980.
Appendix III-B

Constitutional Constraints

on Regulation

Under the checks and balances of our system of

government, the Constitution, as ultimately interpreted by the Supreme Court, requires certain procedural and substantive standards to be met by statutory or other regulation imposed upon an activity.

These requirements depend on the nature of the activity involved. In the present case, it will be useful to

consider first the regulation of basic research and

then the regulation of technological applications,

such as the production of pharmaceuticals by using

genetic engineering methods.

Research

With respect to research, the fundamental question is what limitations, if any, may be placed on the

search for scientific knowledge. The primary applicable constitutional provision is the first amendment,

which has been broadly interpreted by the Supreme

Court to severely limit intrusion by the Government

on all forms of expression.123 Another constitutional

safeguard, known as equal protection, is secondarily

involved.

If the Supreme Court were to recognize a right of

scientific inquiry, its boundaries would not exceed

those for freedom of expression.4 There is disagreement among commentators on this issue concerning

the boundaries of the first amendments and certainly disagreement on the application of generally accepted principles to particular cases. Moreover,

there have been no judicial decisions dealing with the

precise issue at hand. However, it is possible to outline general principles derived from judicial decisions interpreting the first amendment, and indicate

how they might be applied by the courts to attempts

to regulate genetic research.

There are very few limitations on the written or

spoken word. The prohibitions against obscenity or

“fighting words”* clearly would be inapplicable here.

‘Harold P. Green, “The Soundaries of Scientific Freedom” Regulation of

Scientt~ic Inquiry: ticietal Concerns With Research, Keith M. Wulff (cd.)

(Washington, D. C.: AAAS, 1979), pp. 139-143.

aThomas 1. Emerson, “The Constitution and Regulation of Research,” ftegu/ation o~scientt~ic inquiry: Societal Concerns With Reseamh, Keith M. Wulff

(cd.) (Washington, D. C.: AAAS, 1979), pp. 129-137.

‘John A. Robertson, “The Scientists’ Right to Research: A Constitutional

Analysis,” Soufhern Ca/~j’ornia Law Review 5 1:1203, September 1978.

4Green, op. cit., p. 140.

‘Emerson, op. cit., pp. 131-134.

“’Fighting words” are those provoking violent reaction or imminent

disorder.

For many years, the Supreme Court has conceptualized the right of free expression in terms of a marketplace of ideas—through the open and full discussion

of all ideas and related information, the valuable,

valid, or useful ones will be accepted by society,

while the ridiculous or even dangerous ones will be

so demonstrated and discarded. This is a consensual

process; no person, group, or institution has sufficient wisdom to prejudge ideas and deny them

admittance to that intellectual marketplace, even if

they threaten fundamental cultural values, for such

values, if worthwhile, will survive. Under this concept, scientists would certainly have virtually unrestrained freedom to think, speak, and write.

Difficulties arise with actions, such as experimentation, which may be essential to the implementation

of freedom of expression. Recent Supreme Court

cases have recognized a limited protected interest of

the media to gather information as an essential adjunct to freedom of publication. By analogy, it may

be argued that scientists would also be protected in

their research, as a necessary adjunct to freedom of

expression. On the other hand, the information

gathering cases usually involve access to Government facilities, such as courtrooms or prisons. They

are based on the principle that actions by the Government should be open to public scrutiny—a concept not directly applicable to the present issue.

More importantly, the Court has long recognized

that actions related to expression can be regulated

and that regulation may increase with the degree of

the action’s impact on people or the environment.

The Court would probably apply what has been

called a structured balancing test;6 i.e., regulation

would be deemed valid only when the Government

sustains the burden of proving: 1) that there are

“compelling reasons” for the regulation; and 2) that

the objective cannot be achieved by “less drastic

means, ” i.e., by more narrowly drafted regulations

having less impact on first amendment rights.

The second part of the test is fairly straightforward. Governmental restrictions must be kept to a

minimum. E.g., where possible, they should be regulatory rather than prohibitory, temporary rather

than permanent, involve the least burden, and so on.

The difficult part of this test lies in determining

‘Ibid., p. 134.

320
Apppendix III-B—Constitutional Constraints on Regulation ● 321

what is a compelling reason. The protection of health

or the environment is the most clearly acceptable

reason for regulation. In addition, the protection of

individual rights and personal dignity is generally

considered an acceptable reason. E.g., the National

Research Act7 requires that all biomedical and behavioral research involving human subjects supported under the Public Health Service Act be reviewed by an Institutional Review Board in order to

protect the rights and welfare of the subjects.

The above discussion relates to protection from

physical risks due to the process of research. Could

the Government regulate or forbid experimentation

solely because the product (knowledge) threatens

cultural values or other intangibles such as the genetic inheritance of mankind? Religious or philosophical

objections to research, based solely on the rationale

that there are some things mankind should not

know, conflict with the basic principles of freedom

of expression and would not be sufficient reason on

constitutional grounds to justify regulation. Even if

the rationale underlying this objection were expanded to include situations where knowledge threatens

fundamental cultural values about the nature of

man, control of research for such a reason probably

would not be constitutionally permissible. The rationale would again conflict with the marketplace of

ideas concept that is central to freedom of expression. However, what if the knowledge were to provide the means to alter the human species in such a

way that the physical, psychological, and emotional

essence of what it is to be human could be changed?

No precedent exists to provide guidance in determining an answer. Were the situation to arise, the

Supreme Court might fashion another limitation on

the concept of free expression in the same way it

developed the obscenity or “fighting words” doctrines.

The discussion thus far has had as its premise a

direct regulatory approach to research. There is a

more indirect approach, which would be constitutionally permissible and could accomplish much of

what direct regulation might attempt, including prevention of the acquisition of some forms of knowledge. This is the use of the funding power. The

lifeblood of modern science in the United States is

the Federal grant system. Yet it is generally agreed

that Government has no constitutional duty to fund

scientific researches This is a benefit voluntarily provided to which many kinds of conditions may be attached. The only constitutional limitation on such an

approach would be the concept of equal protection—

any restrictions must apply to all or must not be apTPublic Law 93-348 (1974), 42 U.S. C. S289 1-3

‘Green, op. cit., p. 141.

plied in a discriminatory way without compelling

reasons.

Congress could therefore, mandate by law that

certain kinds of research not be funded or be conducted in certain ways. An example is the National

Research Act, discussed previously. However, this

approach may have some serious practical limitations because of the difficulty of determining which

molecular biological research might lead to the proscribed knowledge. Much discretion would have to

be left to the funding agency, which is likely to be unsympathetic or even hostile to such an approach, if it

views its primary mission as fostering research.

Applications and products

Although fears have been expressed that current

genetic technologies may lead to applications that

would be detrimental, no one can reasonably conclude, at the present time, that this will actually occur. For this reason, the most constitutionally permissible approach in all probability will be to regulate the applications of the science. In such situations, whatever harms occur tend to be more tangible and the governmental interests, therefore, more

clearly defined. Moreover, since fundamental constitutional rights are generally not involved, statutes

and regulations are subjected to a lower level of

scrutiny by the Federal courts.

The constitutional authority for Federal regulation

of the applications of technologies such as genetic engineering lies in the commerce clause, article I, section 8 of the Constitution, which grants Congress the

power “To Regulate Commerce with foreign Nations,

and among the Several States. ” In contrast to situations involving fundamental rights, the Supreme

Court has interpreted this clause as giving Congress

extremely broad authority to regulate any activity in

any way connected with commerce. It has been virtually impossible for Congress not to find some connection acceptable to the courts between commerce

and the goals of a particular piece of legislation. * The

standard of review of such legislation by the Federal

courts is to determine if it bears a rational relationship to a valid legislative purpose. If so, the

Court will uphold the legislation and will not second

guess the legislators. This standard of review recognizes that a statute results from the balancing of

competing interests and policies by the branch of

Government created to function in that manner.

*See Wickard v. Filburn, 317 U.S. Ill 0942) in which the Supreme Court upheld civil penalties for violation of acreage allotments established by the

Agricultural Adjustment Act of 1938, covering the amount of wheat that individual farmers could plant, even if the wheat was intended for selfconsumption. The rationale was that even though the individual farmer’s wheat

had no measurable impact on interstate commerce, Congress could properly determine that all wheat of this category, if exempted from regulation,

could undercut the purpose of the Act, which was to increase the price

farmers received for their various crops.
Appendix III-C

Information on International

Guidelines for Recombinant DNA

The following information is based largely on international surveys undertaken by The Committee

on Genetic Experimentation of the International

Council of Scientific Unions reported as of July

1979.’

I. Nations that had established guidelines for conduct

of rDNA research or were using the guidelines of

other nations:

Australia

Belgium

Brazil

Bulgaria

Canada

Czechoslovakia

Denmark

German Democratic

Republic

Federal Republic of

Germany

Finland

France

Hungary

Israel

Italy

Japan

Mexico

Netherlands

New Zealand

Norway

Poland

South Africa

Sweden

Switzerland

Taiwan

United Kingdom

United States

U.S.S.R.

Yugoslavia

II. Nations that had not established guidelines or had

not responded with updated information:

Country Yes No

Austria

Ghana

India

Iran

Jamaica

Korea

Nigeria

Singapore

Sri Lanka

Sudan

Turkey

x

x

x

x

x

x

x

x

x

x

x

111. Nations that had drafted their own guidelines:

Canada Japan

Federal Republic of United Kingdom

‘Report to COGENE ftmm the working gruup on Recombinant DNA Guidelines, May 1980.

Germany United States

France U.S.S.R.

Italy

IV. Nations that had modified the guidelines of other,

indicated, countries:

Australia (UK, U. S.)

Belgium (UK, U. S.)

Brazil (U. S.)

Bulgaria (U. S. S. R., U. S.)

Czechoslovakia (U. S. S. R.,

U. S., Fed. Rep. Ger.)

Denmark (UK)

East German Democratic

Republic (UK, U. S.,

Netherlands)

Finland (U.S. mainly)

Hungary (U. S.)

Mexico (U. S.)

Netherlands (U. S.)

New Zealand

Norway (U. S.)

Poland (U. S.)

South Africa (U. S.)

Sweden (U.S)

Switzerland (U. S.)

Taiwan (U. S., UK)

Yugoslavia

(European Science

Foundation)

V. Nations in which entirely voluntary guidelines have

been adopted:

Finland

VI. Nations with guidelines that are enforceable

through control of research funding:

Australia” Japan

Canada Netherlands

Czechoslovakia Norway

Denmark South Africa

Federal Republic of Sweden

Germany’ Switzerland

France Taiwane

German Democratic United Kingdomf

Republic United States

a’isubmissions may be made directly to the Academy of Science or

through a granting agency. In the latter case, it is a requirement for the applicant to observe the recommendations of the Academy’s Standing Committee if the agency makes a grant for the work. Otherwise, the guidelines

are voluntary with the worker required to make an annual report on progress, or more frequently if conditions of the experiment (such as volumes)

are changed appreciably.”

b’~ntrol through Academy of Sciences and Ministry of Health.”

c~veral research organ~tions require MCeiVers of grants to apply ‘he

NIH guidelines until their own national guidelines are completed.

dThe Netherlands Organization for the Advancement of Pure Research

will only subsidize projects which have been given the committee’s conaent.

e!(waiting for re5pon5e from National Advisory CCJmmittee. ”

f“Notification of proposals to GMAG became compulsory August 1, 1978.

In addition, funding bodies require, as a condition of funding, GMAG’s advice to be sought and followed. ”

322
Appendix I//-C—Information on International Guidelines for Recombinant DNA ● 323

VII. Nations in which guidelines are legally enforceable:

Hungary

U.S.S.R.

Finland. . . . . . . “At present, the guidelines a r e

South Africa. .

entirely voluntary, but in the near

future, the intention is to include

them in the law of infectious diseases when they will become legally

enforceable. ”

“At present the guidelines are not

legally enforceable. They will only

become so if regulations under the

existing Health Act of 1977 and the

Animal Diseases and Parasites Act of

1956 are promulgated; and none are

intended at present. ”

United Kingdom “The regulation to notify GMAG

does not strictly mean that the

Williams Guidelines themselves are

legally enforceable. But, under the

Health and Safety at Work Act

(within which the Regulations were

introduced), it is expected that account will be taken of the relevant

Codes of Practice and the advice

given by GMAG.”

VIII. Nations in which observance of the guidelines is

monitored by a nationally-directed mechanism:

Australia Norway

Czechoslovakia South Africa

German Democratic Sweden

Republic United Kingdom

France United States

Hungary U.S.S.R.

Japan Yugoslavia

IX. Nations in which a license or other authorization

for recombinant DNA activity is granted:

—to an institution: U.S.S.R.

—to an indivdual laboratory: Hungary, Czechoslovakia

—to an individual scientist: Australia, Canada, German Democratic Republic, Federal Republic of

Germany, Finland, France, Japan, Norway, South

Africa, Sweden, United Kingdom”, United States

and U.S.S.R.

Netherlands: “There are gentlemen’s agreements,

signed by the individual scientist, the institution

and the Committee.” The reports of the Committee

also recommend legislation that will require registration of research projects in this field and make

binding the guidelines and supervision of their

observance. (Report of the Committee in Charge of

the Control on Genetic Manipulation, Amsterdam,

March 1977, p. 54.)

Bulgaria, Switzerland: None of the above.

Taiwan: No response.

them in the context of information about the ‘centre’ in which the work is to

go on.”

X. Nations in which special provisions for agriculture

and/or industrial research and applications have

been made:

Czechoslovakia. “10 liter maximum volume of the

culture containing recombinant

DNA”

German

Democratic

Republic. . . . . “The GDR Guidelines will be compulsory for industrial and agricultural applications. 10-liter maximum

deviations may be allowed by the

Minister of Health if suggested by

the Committee. ”

Federal Republic

of Germany . . “Specification of containment of

plants”

France . . . . . . . “Industry, maximum volume of cell

culture is set at 10 liters”

Norway . . . . . . “The Guidelines cover both agriculture and industry. Application of

recombinant DNA research outside

an approved laboratory is prohibited. Otherwise the Committee follows the NIH Guidelines.”

United Kingdom “Agriculture, industry; see Williams

Report, paragraphs 1.3, 2.7, 5.13

and appendix II, section 34. ”

United States . . “Agriculture. NIH Guidelines proU.S.S.R. . . . . . .

Other

respondents, .

vide containment levels for cloning

total plant DNA, plant virus DNA

and plant organelle DNA in E. coli

K-12, and provide general guidance

for the use of plant host-vector systems. 10 liter maximum. A proposed

Supplement to the Guidelines for

voluntary compliance by the private

sector is under consideration by

RAC. Development of a monograph

for large-scale applications has been

proposed.”

“Guidelines are compulsory for industrial and agricultural applications. 10 liter maximum. Deviation

is allowed by the Recombinant DNA

Commission. ”

No
324 ● impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

XI. Number of laboratories currently engaged in recombinant DNA activities:

Country Any labs? How many?

Australia. . . . . . . . . . . .

Austria . . . . . . . . . . . . .

Belgium. . . . . . . . . . . . .

Brazil. . . . . . . . . . . . . . .

Bulgaria. . . . . . . . . ... ,

Canada . . . . . . . . . . . . .

Czechoslovakia ., . . . . .

Denmark. . . . . . . . . . . .

German Democratic

Republic . . . . . . . . . . .

Federal Republic

of Germany. . . . . . . . .

Finland ., . . . . . . . . . . .

France. ., . . . . . . . . . . .

Ghana . . . . . . . . . . . . . .

Hungary . . . . . . . . . . . .

India . . . . . . . . . . . . . . .

Iran. . . . . . . . . . . . . . . .

Israel. . . . . . . . . . . . . . .

Jamaica. . . . . . . . . . . . .

Japan . . . . . . . . . . . . . .

Korea . . . . . . . . . . . . . .

Netherlands . . . . . . . . .

New Zealand. . . . . . . . .

Nigeria . . . . . . . . . . . . .

Norway . . . . . . . . . . . . .

Philippines . . . . . . . . . .

Poland . . . . . . . . . . . . . .

Singapore . . . . . . . . . . .

South Africa . . . . . . . . .

Sri Lanka. . . . . . . . . . . .

Sudan . . . . . . . . . . . . . .

Sweden. . . . . . . . . . . . .

Switzerland. ., . . . . . . .

Taiwan . . . . . . . . . . . . .

Turkey . . . . . . . . . . . . .

United Kingdom . . . . . .

United States.. . . . . . . .

U.S.S.R . . . . . . . . . . . . . .

Yugoslavia. . . . . . . . . . .

yes

noa

yesa

yes

yesa

yes

yes

yesa

yes

yes

yes

yesa

n o

a

yesa

n o

a

n o

a

yesa

n o

a

yes

n o

a

yes

yes

no

yes

n o

a

yesa

n o

a

yesa

n o

a

n o

a

yesa

yesa

yes

n o

a

yes

yesa

yesa

yesb

16

6

5

no response

10-15

3

several

5

10-20

3(3-4 planned)

12

1-2

1

35

7

2

not stated

3

3

2

18

2

45

50

6

4

XII.Countries in which specific training for workers

and safety officers in recombinant DNA activities is

required by the guidelines:

Country Yes No Other

Australia . . . . .

Bulgaria. . . . . .

Canada . . . . . . .

Czechoslovakia.

German

Democratic

Republic . . . . .

Federal Republic

of Germany . .

Finland. .,....

France . . . . . . .

Hungary. . . . . .

Japan. ..., . . .

Netherlands. . .

Norway . . . . . .

South Africa. . .

Sweden . . . . . .

Switzerland . . .

Taiwan. . . . . . .

United Kingdom

United States . .

U.S.S.R. . . . . . .

Yugoslavia . . . .

a

x

x

X

b

x

c

X

d

x

X

e

x

X

g

x

x

x

x

X

f

x

“recommended”

“recommended”

X

h

no response

Other respondents: no or no response to question.

aAustra]ia: URequireexpefii~ through Biosafety COmmiWW.”

bczechoslovakia: ~spific training is recommended.”

c~rman Democratic Republic: “Training courses are organized by the

Committees in cooperation with Akademie fur Arztliche Fortbildungder

DDR.”

dFederal Republic oftkrmany: “Experience as required by law on the

control ofcommunicable diseases.”

eNether]ands: “Thescientistss hould be trained in microbiology.”

fNorway: The Committee certifies training and expertise of personnel are

adequate. ”

gunited I(ingdorn: “Detai]S of training are required; the employer is legally

obliged to provide suitable training. ”

hunited States: ‘“SFific training not required. However, local biohazards

committees are required to certify to the NIH that the training and expertise

of the personnel are adequate. ”

a~wd on replies from previous Questionnaires.

bln preparation.
Appendix III-C—lnformation on international Guidelines for Recombinant DNA ● 325

X111. Countries in which the guidelines are applicable

only to biological agents containing recombinant

DNA, or also cover the recombinant DNA molecules themselves:

Only to

biological Also recombinant

Country agents DNA molecules

Australia. . . . . . . .

Bulgaria . . . . . . . .

Canada. . . . . . . . . (a)

Czechoslovakia . .

German

Democratic

Republic. . . . . . . x

Federal Republic of

Germany. . . . . . . x

Finland. . . . . . . . .

France . . . . . . . . . x

Japan . . . . . . . . . . x

Netherlands . . . . .

New Zealand. . . . .

Norway . . . . . . . .

South Africa. . . . .

Sweden. . . . . . . . .

Switzerland . . . . .

Taiwan. . . . . . . . . x

United Kingdom . .

United States . . . .

U. S. S. R.. . . . . . . . .

x

(a)

x

x

x

x

x

x

x

x

x

X

b

x

a(;uidelines appty to a]i, but containment is not required for naked DNA.

b’The [guidelines-apply to recombinant DNA experiments that are not exempt under Section I-E of the Guidelines. Recombinant DNA molecules that

are not in organisms or viruses are exempt from the Guidelines (I-E-l ).”

XIV. Groups/Committees responsible for carrying out

monitoring of containment procedures:

Country Group

Australia. . . . . . . . Institutional Biosafety Committees.

Bulgaria . . . . . . . . National Committee

Canada . . . . . . . . . “University and Medical Research Council Biohazards Committees”

Czechoslovakia. . . “Under consideration of the National Institutes of Public Health.”

German

Democratic

Republic . . . . . . . “Monitoring is carried out by

local Biosafety Officers, who are

representatives of the Committee

in their institutions. ”

Federal Republic of

Germany. . . . . . . Officers for Biological Safety

monitor the health of employees

and compliance at laboratories;

ZKBS (Zentrale Commission fur

die biologische Sicherheit) has

overall responsibility.

France . . . . . . . . . “Local safety committees”

Hungary. ... , . . . “National Institutes of Public

Health”

Japan ... , . . . . . . “Principal Investigator and Safety

Officer”

Netherlands . . . . . “Site Inspection Commission”

New Zealand. . . . . “Local controlling Committees

are charged with monitoring

observance of Guidelines.

Biological Safety Officers are appointed to take immediate

responsibility.”

Norway . . . . . . . . “Physical containment: Norwegian National Institute of Public

Health. Biological containment:

Committee. ”

South Africa. . . . . “Above P3, Biosafety Committee

of Institute involved and

SAGENE. Below P3, SAGENE

only.”

Sweden. . . . . . . . . Not applicable.

Switzerland . . . . . “At the responsibility of either

the individual investigator or a

local biohazards committee.”

Taiwan. . . . . . . . . No response

United Kingdom . . The Health and Safety Executive

United States . . . . “Observance of containment is to

be monitored by biohazards committees located in institutions in

which the research is conducted.

Effectiveness of containment

procedures is to be monitored by

the principal investigator who is

to report problems to the NIH.”

U. S. S. R.. . . . . . . . . “Local biosafety commission,

State Sanitary Inspection control

group of Recombinant DNA Commission.
326 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

xv. Countries in which the guidelines apply to all gene

combinations instructed by cell-free methods, or

only to molecules containing combinations of

genes from different species:

Molecules

All gene com- containing

binations con- combinations of

structed by cell- genes from

Country free methods different species

Australia. . . . . . . .

Canada. . . . . . . . .

Czechoslovakia. . .

German

Democratic

Republic. . . . . . .

Federal Republic of

Germany. . . . . . .

Finland. . . . . . . . .

France . . . , . . . . .

Japan . . . . . . . . . .

Netherlands . . . . .

New Zealand. . . . .

Norway . . . . . . . .

South Africa . . . . .

Sweden. . . . . . . . .

Switzerland . . . . .

Taiwan. . . . , . . . .

United Kingdom . .

United States . . . .

U. S. S. R.. . . . . . . . .

x

x

x

x

X

a

x

x

x

X

b

x

x

x

x

x

x

aFederal Republic of Germany Selfwloning experiments involving nonpathogenic donors and hosts shall be reported to ZKBS.

bNetherlands “The definition of recombinant DNA has recedy been

modified and includes the insertion of chemically synthesized DNA molecules into a vector. ”

cunited Kingdom “The G~u@s provision] interpretation Of their Own remit is that they are concerned with work involving genetic manipulation,

defined for these purposes as: the formation of new combinations of heritable materials by the insertion of nucleic acid molecules, produced by

whatever means outside the cell, into any virus, bacterial plasmid, or other

vector sbystem so as to allow their incorporation into a host organism in

which they do not naturally occur but in which they are capable of continued propagation.”

XVI. Countries in which the guidelines restrict the intentional dissemination into the environment of

biological agents containing recombinant DNA:

All respondents . .

Australia. . . . . . . .

German

Democratic

Republic . . . . . . .

Yesa

Not explicity so

“Exceptions have to be discussed

by the Committee and require

special permission by the Minister of Health.”

New Zealand. . . . . “Yes, with the approval of the

National Committee.”

United Kingdom . . “The question has not arisen.”

Other respondents No

aAr@ there anv circumstances under which such dissemination can be carried out?

XVII. Countries in which the guidelines are restricted

to recombinant DNA activities or also cover

other areas of genetic experimentation:

Recombinant Other areas of

DNA genetic

Country activities experimentation

Australia. ... , ., .

Bulgaria . . . . . . . .

Canada . . . . . . . . .

Czechoslovakia. . .

German

Democratic

Republic . . . . . . .

Federal Republic of

Germany. . . . . . .

Finland. . . . . . . . .

France . . . . . . . . .

Hungary. . . . . . . .

Japan . . . . . . . . . .

Netherlands . . . . .

New Zealand. . . . .

Norway . . . . . . . .

South Africa. . . . .

Sweden. . . . . . . . .

Switzerland ... , .

United Kingdom . .

United States . . . .

U. S. S. R.. . . . . . . . .

x

a

x

X

b

x

x

x

x

x

x

x

x

x

x

x

x

x

x

X

C

X

d

a“At present, the terms of reference of the Academy Committee refer

only to in vitru experiments (i.e., the use of restriction enzymes and ligases).

An ad hoc Academy Committee is about to investigate in vivo experimentation, with the following terms of reference:

1. Examine whether, other than by using the technique of in vitru recombinant DNA construction, new hybrid nucleic acid molecules can be

produced that are potentially dangerous to humans, animals, or plants.

In so doing, the committee should give particular attention to the following possibilities:

—The use of mixed infections involving human or animal viruses, or the

use of bacteria or fungi.

–The introduction of foreign DNA into plants and the production of

new plant pathogens.

2. Consider whether there are certain classes of viral pathogens (e.g.,

polio) on which experimentation should not be carried out unless a special need is demonstrated.” ‘

b“work with animal viruses and cells”

C“i.e., cell fusion with approval of National Committee”

d“other c]osely related areas are also covered.”
Appendix //l-C—Information on International Guidelines for Recombinant DNA . 327

XVIII. Countries in which the recombinant DNA advisory committee includes public representatives as well as scientists:

Country Yes No

Australia. . . . . . . .

Bulgaria . . . . . . . .

Canada . . . . . . . . .

Czechoslovakia. . .

Denmark . . . . . . .

German

Democratic

Republic . . . . . . .

Federal Republic of

Germany. .,

Finland. . . . .

France . . . .,

Hungary. . . .

Italy . . . . . . .

Japan . . . . . .

Netherlands .

New Zealand.

Norway . . . .

South Africa .

Sweden. . . . .

Switzerland .

Taiwan. . . . .

. . . .

. . . .

. . . .

. . . .

... ,

. . . .

.,. .

. . . .

. . . .

. . . .

. . . .

. . . .

. . . .

United Kingdom . .

United States . . . .

U. S. S. R.. . . . . . . . .

x

x

x

x

x

Finland. . . . . . . . .

x

x

x

x

x

x

x

x

x

x

x

x

x

x

x

x

x

Composition of DNA advisory committees is as follows:

Australia. . . . . . . .

Canada . . . . . . . . .

Czechoslovakia. . .

Denmark . . . . . . .

German

Democratic

Republic . . . . . . .

8 scientists

5 laymen (1 lawyer, 1 businessman, 3 generalists); 6 scientists (2

M.D.s, 3 virologists/cancer specialists, 1 recombinant DNA specialist)

6 members representing molecular biology, genetics, microbiology, medicine

9 scientists and administrative

representatives.

3 geneticists, 1 biochemist, 2 bacteriologists, 2 virologist, 1 jurist,

1 representative of trade union

of GDR.

Federal Republic of

Germany. . . . . . . 4 experts working in the field of

recombinant DNA research; 4 experts who, though not working

in the field of recombinant DNA

France . . . . . . . . .

Hungary. , . . . . . .

Italy . . . . . . . . . . .

Japan . . . . . . . . . .

Netherlands . . . . .

New Zealand. . . . .

Norway . . . . . . . .

South Africa . . . . .

Sweden. . . . . . . . .

research, possess specific knowledge in the implementation of

safety measures in biological research work, particularly however in microbiology, cytobiology, or hygiene and, in addition, 4

outstanding individuals, for example from the trade unions, industry, and the research-promoting organizations.

27 members: 6 molecular biology, 3 genetics, 3 microbiology, 1

virology, 1 plant physiology, 3 infectious diseases, 3 epidemiology,

2 enteric bacteria, 1 cell cultures,

3 public health, 1 occupational

health.

13 members, 4 observers, 1 secretary

Scientists

8 molecular biologists, 4 microbiologists, 1 civil servant (Health

Ministry).

(Combines both Steering Committee and Advisory Group): 7 recombinant DNA scientists, 7 scientists in other fields, 6 specialists in medicine and biohazards, 2

lawyers, 2 specialists in physical

containment, 3 public representatives.

14 scientists representing genetics, molecular biology, bacteriology, virology, botany, medicine,

ethics and social aspects of health

and health-care. To be added: a

committee composed of scientists

and representatives of industry

and trade unions.

1 molecular biologist, 1 microbial

geneticist, 1 virologist, 1 botanist

(molecular biologist), 1 human

geneticist (medically qualified).

3 biochemists, 2 medicine, 1 veterinary medicine, 1 lawyer, 1

artist.

One each from: Council for Scientific and Industrial Research,

Medical Research Council, Department of Health, Department

of Agricultural Technical Services. Three from universities,

public and legal professions.

No response
328 ● Impacts of Applied Genetics— Micro-Organisms, Plants, and Animals

Switzerland ... , . 12 members representing medi- ogy: 2, Plant Genetics: 2, Law: 2,

cine, microbiology, molecular Environmental Concerns, Labbiology, antibiotics, industry, oratory Technician, Infectious

university management, and 7 Diseases, Occupational Health,

governmental departmental Education: 1 each.

assessors. U. S. S. R.. .. ......8 scientists

United States . . . . Molecular biology: 6, Molecular Yugoslavia .. ....3 geneticists

Genetics: 5, Ethics: 3, Microbiol-
Appendix IV

Planning Workshop Participants,

Other Contractors and Contributors,

and Acknowledgments

Planning workshop participants

Philip Bereano

University of Washington

Ralph Hardy

E. L Du Pent de Nemours & Co., Inc.

Patricia King

Georgetown University Law Center

Charles Lewis

U.S. Department of Agriculture

Herman Lewis

National Science Foundation

Pamela Lippe

Friends of the Earth

Victor McKusick

Johns Hopkins University Medical School

Elena O. Nightingale

Institute of Medicine

Gilbert Omenn

Office of Science and Technology Policy

Donna Parratt

Congressional Research Service

Walter Shropshire

Smithsonian Radiation Biology Laboratory

Leroy Walters

The Kennedy Institute

Other contractors and contributors

Betsy Amin-Arsala

Richard J. Auchus

Massachusetts Institute of Technology

Fred Bergmann

National Institutes of Health

Charles L. Cooney

Massachusetts Institute of Technology

Richard Curtin

Robert A. Cuzick

Massachusetts Institute of Technology

Roslyn Dauber

Arnold L. Demain

Massachusetts Institute of Technology

Richard B. Emmitt

F. Eberstadt & Co.

Emanuel Epstein

University of California, Davis

Robert F. Fleischaker

Massachusetts Institute of Technology

Odelia Funke

George E. Garrison

F. Eberstadt & Co.

Reinaldo F. Gomez

Massachusetts Institute of Technology

John Hamilton

Neal F. Jensen

Scott A. King

F. Eberstadt & Co.

Harvey Lodish

Massachusetts Institute of Technology

L. D. Nyhart

Massachusetts Institute of Technology

ChoKyun Rha

Massachusetts Institute of Technology

William Scanlon

Andrew Schmitz

University of California, Berkeley

Michael J. Shodell

The Sterling-Hobe Corp.

David Tse

Massachusetts Institute of Technology

James Welsh

Montana State University

George Whiteside

Massachusetts Institute of Technology

Bernard Wolnak and Associates

Nancy Woods

OTA intern

Acknowledgments

A large number of individuals provided valuable

advice and assistance to OTA during this assessment.

In particular, we would like to thank the following

people:

John Adams

Pharmaceutical Manufacturers Association

Rupert Amann

Colorado State University

William Amen, Jr.

Cetus Corp.

Daniel Azarnoff

Searle Laboratories

A. L. Barr

West Virginia University

K. J. Betteridge

Animal Diseases Research Institute

329
330 ● Impacts of Applied Genetics—Micro-Organisms, Plants, and Animals

Jerome Birnbaum

Merck, Sharp & Dohme Research Laboratories

Gerald Bjorge

U.S. Patent and Trademark Office

Hugh Bollinger

Plant Resources Institute

G. Eric Bradford

University of California

Robert Brackett

Parke, Davis & Co.

Robert Byrnes

Genentech, Inc.

Daniel Callahan

The Hastings Center

Alexander M. Capron

President’s Commission for the Study of Ethical

Problems in Medicine and Biomedical and

Behavioral Research

Robert Church

University of Calgary

H. Wallis Clark

University of California

Gail Cooper

Environmental Protection Agency

Joseph P. Dailey

Revlon Health Care Group

Frank Dickinson

Agricultural Research Center

Donald R. Dunner

Finnegan, Henderson, Farabow, Garrett &

Dunner

Roger B. Dworkin

Indiana University School of Law

Richard P. Elander

Bristol-Myers Co.

Peter Elsden

Colorado State University

Haim Erder

University of Pennsylvania

James F. Evans

Pennsylvania Embryo Transfer Service

Kenneth Evans

Plant Variety Protection Office

Richard Faust

Hoffman-La Roche, Inc.

Herman Finke

Sterling Systems

Robert H. Foote

Cornell University

Orrie M. Friedman

Collaborative Research, Inc.

William J. Gartland, Jr.

National Institutes of Health

Kenneth Goertzen

Seed Research, Inc.

Michael Goldberg

Food and Drug Administration

Maxwell Gordon

Bristol Laboratories

Lorance L. Greenlee

Keil and Witherspoon

Ralph Hardy

E. I. Du Pent de Nemours and Co,, Inc.

W. C. D. Hare

Animal Diseases Research Institute

Paul Harvey

U.S. Department of Agriculture

Harold W. Hawk

Agricultural Research Center

Richard L. Hinman

Pfizer, Inc.

Peter Barton Hutt

Covington & Burling

E. Keith Inskeep

West Virginia University

Irving Johnson

tiny Research Laboratories

Charles Kiddy

Agricultural Research Center

Thomas D. Kiley

Genentech, Inc.

Carole Kitti

National Science Foundation

Duane C. Kraemer

Texas A&M University

Sheldon Krimsky

Tufts University

W. W. Lampeter

Lehn und Versuchtsgut

Earl Lasley

Monsanto Farmers Hybrid

F. Douglas Lawrason

Schering Corp.

Bernard Leese

Plant Variety Protection Office

Stanley Leibo

Oak Ridge National Laboratory

Morris Levin

Environmental Protection Agency

Herman Lewis

National Science Foundation

Peter Libassi

Verner, Lipfert, Bernhard & McPherson

Paul J. Luckern

U.S. Department of Justice

Clement Markert

Yale University

Ralph R. Maurer

U.S. Meat Animal Research Center
Appendix IV—Planning Workshop Participants, Other Contractors and Contributors, and Acknowledgments ● 331

Robert McKinnell

University of Minnesota

Edward Mearns, Jr.

Case Western Reserve University School

of Medicine

Alan S. Michaels

Stanford University

Elizabeth Milewski

National Institutes of Health

Henry I, Miller

Food and Drug Administration

Paul Miller

W. R. Grace

A. V. Nalbandov

University of Illinois

Claude H. Nash

Smith Kline & French Laboratories

Dorothy Nelkin

Resources for the Future

Gordon Niswender

Colorado State University

Elena Ottolenghi-Nightingale

National Academy of Medicine

David Padwa

Agrigenetics, Inc.

Seth Pauker

National Institute of Occupational Safety

& Health

J. B. Peters

West Virginia University

Nancy Pfund

Stanford University School of Medicine

James Punch

The Upjohn Co.

Neils Reimers

Stanford University

Ira Ringler

Abbott Laboratories

Roman Saliwanchik

The Upjohn Co.

Robert B. Samuels

Beckman Instruments

George E. Seidel, Jr.

Colorado State University

Sarah M. Seidel

Colorado State University

Thomas J. Sexton

U.S. Department of

Brian F. Shea

Ralph Silber

Stanford University

Elizabeth L. Singh

Charles G. Smith

Agriculture

Revlon Health Group

Animal Diseases Research Institute

Davor Solter

Wistar Institute of Anatomy and Biology

Mark Sorrells

Cornell University

G. F. Sprague

University of Illinois

Richard Staples

Cornell University

Gerald G. Still

U.S. Department of Agriculture

Charles W. Stuber

North Carolina State University

Bernard Talbot

National Institutes of Health

Rene Tegtmeyer

U.S. Patent and Trademark Office

Clair E. Terrill

U.S. Department of Agriculture

Robin Tervit

Colorado State University

Stephen Turner

Bethesda Research Laboratories, Inc.

L. D. Van Vleck

Cornell University

Robert Walton

W. R. Grace

Charles Weiner

Massachusetts Institute of Technology

Ray W. Wright, Jr.

Washington State University

Susan Wright

University of Michigan

Oskar R. Zaborsky

National Science Foundation

M. S. Zuber

University of Missouri

o