Federal Register / Vol. 77, No. 172 / Wednesday, September 5, 2012 / Proposed Rules 54499 

and Local Notice to Mariners. Such 

notifications will include the date and 

time that enforcement is suspended as 

well as the date and time that 

enforcement will resume. 

(3) Violations of this regulated 

navigation area should be reported to 

the COTP, at 203–468–4401 or on VHF– 

Channel 16. Persons in violation of this 

regulated navigation area may be subject 

to civil or criminal penalties. 

Dated: August 22, 2012. 

D.B. Abel, 

Rear Admiral, U.S. Coast Guard, Commander, 

First Coast Guard District. 

[FR Doc. 2012–21760 Filed 9–4–12; 8:45 am] 

BILLING CODE 9110–04–P 

ENVIRONMENTAL PROTECTION 

AGENCY 

40 CFR Part 725 

[EPA–HQ–OPPT–2011–0740; FRL–9348–1] 

RIN 2070–AJ65 

Microorganisms; General Exemptions 

From Reporting Requirements; 

Revisions to Recipient Organisms 

Eligible for Tier I and Tier II 

Exemptions 

AGENCY: Environmental Protection 

Agency (EPA). 

ACTION: Proposed rule. 

SUMMARY: EPA received petitions to add 

Trichoderma reesei and Bacillus 

amyloliquefaciens to the list of 

microorganisms that may be used as 

recipient microorganisms in order to 

qualify for the exemption from full 

notification and reporting procedures 

under the Toxic Substances Control Act 

(TSCA) for new microorganisms that are 

being manufactured for introduction 

into commerce. Based on EPA’s 

evaluation of these petitions, EPA has 

made a preliminary determination that 

certain strains of both microorganisms 

will not present an unreasonable risk of 

injury to health or the environment 

when used as a recipient microorganism 

provided that certain criteria for the 

introduced genetic material and the 

physical containment conditions are 

met. Therefore, EPA is proposing to add 

two additional microorganisms to the 

list of recipient microorganisms that are 

eligible for exemptions from full 

reporting for the manufacture (including 

import) of new microorganisms. 

DATES: Comments must be received on 

or before November 5, 2012. 

You may submit a request for an 

opportunity to present oral comments in 

writing on or before October 5, 2012, 

and if a written request is received by 

EPA, an informal public hearing will be 

held on this proposed rule in 

Washington, DC. For further 

information on the informal public 

hearing, see Unit I.C. 

ADDRESSES: Submit your written request 

for an opportunity to present oral 

comments, identified by docket 

identification (ID) number EPA–HQ– 

OPPT–2011–0740, to the mailing or 

hand delivery addresses in this unit. 

Submit your comments, identified by 

docket ID number EPA–HQ–OPPT– 

2011–0740, by one of the following 

methods: 

• Federal eRulemaking Portal: http:// 

www.regulations.gov. Follow the online 

instructions for submitting comments. 

• Mail: Document Control Office 

(7407M), Office of Pollution Prevention 

and Toxics (OPPT), Environmental 

Protection Agency, 1200 Pennsylvania 

Ave. NW., Washington, DC 20460–0001. 

• Hand Delivery: OPPT Document 

Control Office (DCO), EPA East Bldg., 

Rm. 6428, 1201 Constitution Ave., NW., 

Washington, DC. Attention: Docket ID 

Number EPA–HQ–OPPT–2011–0740. 

The DCO is open from 8 a.m. to 4 p.m., 

Monday through Friday, excluding legal 

holidays. The telephone number for the 

DCO is (202) 564–8930. Such deliveries 

are only accepted during the DCO’s 

normal hours of operation, and special 

arrangements should be made for 

deliveries of boxed information. 

Instructions: Direct your comments to 

docket ID number EPA–HQ–OPPT– 

2011–0740. EPA’s policy is that all 

comments received will be included in 

the docket without change and may be 

made available online at http:// 

www.regulations.gov, including any 

personal information provided, unless 

the comment includes information 

claimed to be Confidential Business 

Information (CBI) or other information 

whose disclosure is restricted by statute. 

Do not submit information that you 

consider to be CBI or otherwise 

protected through regulations.gov or 

email. The regulations.gov Web site is 

an ‘‘anonymous access’’ system, which 

means EPA will not know your identity 

or contact information unless you 

provide it in the body of your comment. 

If you send an email comment directly 

to EPA without going through 

regulations.gov, your email address will 

be automatically captured and included 

as part of the comment that is placed in 

the docket and made available on the 

Internet. If you submit an electronic 

comment, EPA recommends that you 

include your name and other contact 

information in the body of your 

comment and with any disk or CD–ROM 

you submit. If EPA cannot read your 

comment due to technical difficulties 

and cannot contact you for clarification, 

EPA may not be able to consider your 

comment. Electronic files should avoid 

the use of special characters, any form 

of encryption, and be free of any defects 

or viruses. 

Docket: All documents in the docket 

are listed in the docket index available 

at http://www.regulations.gov. Although 

listed in the index, some information is 

not publicly available, e.g., CBI or other 

information whose disclosure is 

restricted by statute. Certain other 

material, such as copyrighted material, 

will be publicly available only in hard 

copy. Publicly available docket 

materials are available electronically at 

http://www.regulations.gov, or, if only 

available in hard copy, at the OPPT 

Docket. The OPPT Docket is located in 

the EPA Docket Center (EPA/DC) at Rm. 

3334, EPA West Bldg., 1301 

Constitution Ave. NW., Washington, 

DC. The EPA/DC Public Reading Room 

hours of operation are 8:30 a.m. to 4:30 

p.m., Monday through Friday, excluding 

legal holidays. The telephone number of 

the EPA/DC Public Reading Room is 

(202) 566–1744, and the telephone 

number for the OPPT Docket is (202) 

566–0280. Docket visitors are required 

to show photographic identification, 

pass through a metal detector, and sign 

the EPA visitor log. All visitor bags are 

processed through an X-ray machine 

and subject to search. Visitors will be 

provided an EPA/DC badge that must be 

visible at all times in the building and 

returned upon departure. 

FOR FURTHER INFORMATION CONTACT: For 

technical information contact: Brian 

Lee, Chemical Control Division 

(7405M), Office of Pollution Prevention 

and Toxics, Environmental Protection 

Agency, 1200 Pennsylvania Ave., NW., 

Washington, DC 20460–0001; telephone 

number: (202) 564–6293; email address: 

lee.brian@epa.gov. 

For general information contact: The 

TSCA-Hotline, ABVI-Goodwill, 422 

South Clinton Ave., Rochester, NY 

14620; telephone number: (202) 554– 

1404; email address: TSCAHotline@epa.gov. 

SUPPLEMENTARY INFORMATION: 

I. General Information 

A. Does this action apply to me? 

You may be potentially affected by 

this action if you produce, import, 

process, or use either intergeneric 

Trichoderma reesei or intergeneric 

Bacillus amyloliquefaciens. Potentially 

affected entities may include, but are 

not limited to: 


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• Basic Chemical Manufacturing 

(NAICS code 3251). 

• Pesticide, Fertilizer and other 

Agricultural Chemical manufacturing 

(NAICS code 3253). 

• Other Chemical Product and 

Preparation Manufacturing (NAICS code 

3259). 

This listing is not intended to be 

exhaustive, but rather provides a guide 

for readers regarding entities likely to be 

affected by this action. Other types of 

entities not listed in this unit could also 

be affected. The North American 

Industrial Classification System 

(NAICS) codes have been provided to 

assist you and others in determining 

whether this action might apply to 

certain entities. If you have any 

questions regarding the applicability of 

this action to a particular entity, consult 

the technical person listed under FOR

FURTHER INFORMATION CONTACT. 

B. What should I consider as I prepare 

my comments for EPA? 

1. Submitting CBI. Do not submit this 

information to EPA through 

regulations.gov or email. Clearly mark 

the part or all of the information that 

you claim to be CBI. For CBI 

information in a disk or CD–ROM that 

you mail to EPA, mark the outside of the 

disk or CD–ROM as CBI and then 

identify electronically within the disk or 

CD–ROM the specific information that 

is claimed as CBI. In addition to one 

complete version of the comment that 

includes information claimed as CBI, a 

copy of the comment that does not 

contain the information claimed as CBI 

must be submitted for inclusion in the 

public docket. Information so marked 

will not be disclosed except in 

accordance with procedures set forth in 

40 CFR part 2. 

2. Tips for preparing your comments. 

When submitting comments, remember 

to: 

i. Identify the document by docket ID 

number and other identifying 

information (subject heading, Federal 

Register date and page number). 

ii. Follow directions. The Agency may 

ask you to respond to specific questions 

or organize comments by referencing a 

Code of Federal Regulations (CFR) part 

or section number. 

iii. Explain why you agree or disagree; 

suggest alternatives and substitute 

language for your requested changes. 

iv. Describe any assumptions and 

provide any technical information and/ 

or data that you used. 

v. If you estimate potential costs or 

burdens, explain how you arrived at 

your estimate in sufficient detail to 

allow for it to be reproduced. 

vi. Provide specific examples to 

illustrate your concerns and suggest 

alternatives. 

vii. Explain your views as clearly as 

possible, avoiding the use of profanity 

or personal threats. 

viii. Make sure to submit your 

comments by the comment period 

deadline identified. 

C. Can I request an opportunity to 

present oral comments to the agency? 

You may submit a request for an 

opportunity to present oral comments. 

This request must be made in writing 

and be identified by docket ID number 

EPA–HQ–OPPT–2011–0740. This 

written request must be submitted to the 

mailing or hand delivery addresses 

provided under ADDRESSES. If such a 

request is received on or before October 

5, 2012, EPA will hold an informal 

public hearing on this proposed rule in 

Washington, DC. If such a request is 

received, EPA will announce the 

scheduling of the informal public 

hearing in a subsequent document in 

the Federal Register. If an informal 

public hearing is announced, and if you 

are interested in attending or presenting 

oral and/or written comments at the 

informal public hearing, you should 

follow the instructions provided in the 

subsequent Federal Register document 

announcing the informal public hearing. 

II. Background 

A. What action is the agency taking? 

EPA received petitions to add 

Trichoderma reesei and Bacillus 

amyloliquefaciens to the list of recipient 

microorganisms at § 725.420 that are 

eligible for the regulatory exemptions 

applicable to new microorganisms that 

are manufactured for introduction into 

commerce (Refs. 1–3). EPA has made a 

preliminary determination that both of 

the microorganisms, with certain 

limitations, meet the criteria for 

addition to the list—i.e., they will not 

present an unreasonable risk of injury to 

health or the environment provided that 

the other conditions of the exemptions 

at 40 CFR part 725, subpart G, relating 

to the introduced genetic material, and 

the physical containment of the new 

microorganisms, have been met. 

Therefore, this document proposes to 

grant the exemption petition for these 

two microorganisms. 

EPA is proposing to restrict the 

exemption for Trichoderma reesei to the 

Trichoderma reesei strain QM6a and its 

derivatives (hereafter, T. reesei QM6a). 

In addition, EPA is proposing to restrict 

the T. reesei QM6a exemption to use 

under submerged standard industrial 

fermentation conditions; as described in 

this proposed rule, these conditions are 

typical throughout industry and would 

also meet the existing physical 

containment and control requirements 

for the tiered exemptions under 

§ 725.422. EPA would also restrict the T. 

reesei QM6a exemption to fermentation 

operations in which no solid plant 

material or insoluble substrate is present 

in the fermentation broth. EPA is also 

proposing to require that any 

fermentation of solid plant material or 

insoluble substrate may only be 

initiated after the inactivation of T. 

reesei QM6a by a procedure that meets 

the existing requirements in 

§ 725.422(d), i.e., by a procedure that 

has been demonstrated and documented 

to be effective in reducing the viable 

microbial population by at least 6 logs. 

Additionally, EPA is proposing to 

limit the exemption for B. 

amyloliquefaciens to only industrial 

strains of Bacillus amyloliquefaciens 

that would fall into the subspecies 

Bacillus amyloliquefaciens subsp. 

amyloliquefaciens (hereafter, B. 

amyloliquefaciens). 

B. What is the agency’s legal authority 

for taking this action? 

This action is being taken under the 

authority of TSCA section 5(h)(4) (15 

U.S.C. 2604(h)(4)). 

Section 5(a)(1) of TSCA requires that 

persons notify EPA at least 90 days 

before they manufacture (the term 

‘‘manufacture’’ includes import under 

TSCA) for commercial purposes a 

‘‘new’’ chemical substance, or 

manufacture (including import) or 

process a chemical substance for a 

‘‘significant new use.’’ TSCA defines 

‘‘chemical substance’’ broadly and in 

terms that cover intergeneric 

microorganisms as well as traditional 

chemical substances. Therefore, for the 

purposes of TSCA, a ‘‘new 

microorganism,’’ like a ‘‘new chemical 

substance,’’ is one that is not listed on 

the TSCA Chemical Substances 

Inventory (TSCA Inventory) compiled 

under TSCA section 8(b). Section 5(h)(4) 

of TSCA authorizes EPA, upon 

application and by rule, to exempt the 

manufacturer or importer of any new 

chemical substance from part or all of 

the provisions of TSCA section 5, if EPA 

determines that the manufacture, 

processing, distribution in commerce, 

use, or disposal of the new chemical 

substance will not present an 

unreasonable risk of injury to human 

health or the environment. 

C. Existing EPA Regulatory 

Requirements and Exemption Standard 

Manufacturers are required to report 

certain information to EPA 90 days 


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before commencing the manufacture of 

intergeneric microorganisms that are not 

listed on the TSCA Inventory. EPA 

regulations at 40 CFR part 725 establish 

the mechanisms for reporting this 

information. 

Any manufacturer of a living 

intergeneric microorganism who is 

required to report under TSCA section 

5 must file a Microbial Commercial 

Activity Notice (MCAN) with EPA, 

unless the activity is eligible for one of 

the specific exemptions. The general 

procedures for filing MCANs are 

described in 40 CFR part 725, subpart B. 

EPA regulations establish two 

exemptions for new microorganisms, 

after the research and development 

stage, which are being manufactured for 

introduction into commerce: The Tier I 

and Tier II exemptions. 

Under the Tier I exemption, if three 

criteria are met, manufacturers are only 

required to notify EPA that they are 

manufacturing a new microorganism 

that qualifies for this exemption 10 days 

before commencing manufacture, and to 

keep certain records. 40 CFR 725.400. 

To qualify for the Tier I exemption, a 

manufacturer must use one of the 

recipient organisms listed in § 725.420, 

and must implement specific physical 

containment and control technologies. 

In addition, the genetic material 

introduced into the recipient 

microorganism must be wellcharacterized, limited in size, poorly 

mobilizable, and free of certain 

sequences. 40 CFR 725.421. 

A manufacturer who otherwise meets 

the conditions of the Tier I exemption 

may modify the specified containment 

restrictions, but must submit a Tier II 

exemption notification. 40 CFR 725.428. 

The Tier II exemption requires 

manufacturers to submit an abbreviated 

notification describing the modified 

containment, and provides for a 45 day 

period, during which EPA would review 

the proposed containment. 40 CFR 

725.450 and 725.470. The manufacturer 

may not proceed under this exemption 

until EPA approves the exemption. 40 

CFR 725.470. 

EPA established a petition process at 

§ 725.67 to provide a mechanism for the 

public to propose additional 

microorganisms as candidates for the 

tiered exemptions. 

Section 725.67 directs a petitioner to 

submit information to demonstrate that 

‘‘any activities affected by the requested 

exemption will not present an 

unreasonable risk of injury to health or 

the environment.’’ 40 CFR 725.67(a)(2). 

In addition, a petitioner is responsible 

to provide supporting information for 

this determination in four general 

categories: 

1. The effects of the new 

microorganism on health and the 

environment. 

2. The magnitude of exposure of 

human beings and the environment to 

the new microorganism. 

3. The benefits of the new 

microorganism for various uses and the 

availability of substitutes for such uses. 

4. The reasonably ascertainable 

economic consequences of granting or 

denying the petition, including effects 

on the national economy, small 

business, and technological innovation. 

Section 725.67 also specifies that 

when applying to list a recipient 

microorganism for the tiered exemption 

under § 725.420, petitioners should 

include information addressing six 

specified criteria, which EPA will use to 

evaluate the microorganism for listing. 

40 CFR 725.67(a)(3)(iii). The six criteria 

are: 

• Identification and classification of 

the microorganism using available 

genotypic and phenotypic information. 

• Information to evaluate the 

relationship of the microorganism to 

any other closely related 

microorganisms which have a potential 

for adverse effects on health or the 

environment. 

• A history of safe commercial use for 

the microorganism. 

• Commercial uses indicating that the 

microorganism products might be 

subject to TSCA. 

• Studies which indicate the 

potential for the microorganism to cause 

adverse effects to health or the 

environment. 

• Studies which indicate the survival 

characteristics of the microorganism in 

the environment. 

III. EPA’s Evaluation of Available 

Information on the Proposed 

Microorganisms for the Criteria 

Delineated in § 725.67 

Pursuant to § 725.67, Genencor 

International, Inc., (subsequently 

supported by the Enzyme Technical 

Association (ETA)) and Novozymes 

North America, Inc., submitted Letters 

of Application to EPA requesting that 

Trichoderma reesei and Bacillus 

amyloliquefaciens (Refs. 1 and 2) be 

added to § 725.420 as candidate 

recipient microorganisms for the tiered 

exemptions. The letters of application 

provided information that the 

submitters believed demonstrate that 

activities affected by the requested 

exemptions would not present an 

unreasonable risk of injury to health or 

the environment. Information regarding 

the criteria specified in §§ 725.67(a)(2) 

and 725.67(a)(3)(iii) were addressed in 

these letters of application to list 

Trichoderma reesei and Bacillus 

amyloliquefaciens as recipient 

microorganisms under § 725.420. 

EPA has made a preliminary 

determination based on the information 

provided in the Letters of Application 

(Refs. 1 and 2), supplemental 

information provided by ETA (Refs. 4 

and 5), and other information available 

to EPA that T. reesei QM6a, with certain 

restrictions, and B. amyloliquefaciens 

will not present an unreasonable risk of 

injury to health or the environment 

when used as a recipient microorganism 

provided the existing criteria for the 

introduced genetic material and for 

physical containment conditions at 

§ 725.422 are met. EPA’s Risk 

Assessments for these two 

microorganisms (Refs. 6 and 7) are 

available in the docket. This unit 

presents a summary of EPA’s evaluation 

of the available information pertinent to 

the six criteria delineated in 

§ 725.67(a)(3)(iii) for both 

microorganisms. These criteria follow: 

• Identification and classification of 

the microorganism using available 

genotypic and phenotypic information. 

• Information to evaluate the 

relationship of the microorganism to 

any other closely related 

microorganisms that have a potential for 

adverse effects on health or the 

environment. 

• A history of safe commercial use for 

the microorganism. 

• Commercial uses indicating that the 

microorganism products might be 

subject to TSCA. 

• Studies which indicate the 

potential for the microorganism to cause 

adverse effects to health or the 

environment. 

• Studies which indicate the survival 

characteristics of the microorganism in 

the environment. 

Units V. and VI. summarize EPA’s 

evaluation of the information relating to 

the criteria delineated in § 725.67(a)(2) 

that address hazard, exposure, benefits, 

and economic consequences. 

Specifically: 

• The effects of the new 

microorganism on health and the 

environment. 

• The magnitude of exposure of 

human beings and the environment to 

the new microorganism. 

• The benefits of the new 

microorganism for various uses and the 

availability of substitutes for such uses. 

• The reasonably ascertainable 

economic consequences of granting or 

denying the exemption, including 

effects on the national economy, small 

business, and technological innovation. 

Unit V. provides a summary of EPA’s 

assessments of the risks to health and 


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the environment for both 

microorganisms. EPA’s Risk Assessment 

documents (Refs. 6 and 7) provide more 

detailed information, and supporting 

references, for EPA’s evaluation of the 

available information and the potential 

risks to health and the environment. 

Unit VI. provides a summary of EPA’s 

assessments of the economic benefits 

and consequences of adding both 

microorganisms to § 725.420. 

A. Evaluation of Available Information 

Relevant to the Criteria at § 725.67 for 

T. reesei QM6a as a Recipient 

Microorganism With Specified 

Conditions of Growth 

1. Identification and classification of 

the microorganism using available 

genotypic and phenotypic information. 

T. reesei is a fungus originally isolated 

in the Solomon Islands in 1944. T. 

reesei is a hypercellulolytic fungus 

found on deteriorating military fabrics 

such as tents and clothing. This isolate, 

designated as QM6a, was initially 

named Trichoderma viride. 

Approximately 20 years later, QM6a 

was re-classified as Trichoderma reesei. 

Trichoderma reesei is the species 

name given to the anamorphic form 

(this form reproduces asexually) of the 

fungus whose telemorphic form (this 

form reproduces sexually) is now 

understood to be Hypocrea jecorina. 

Recent taxonomic studies have shown 

that the species T. reesei consists only 

of this single isolate QM6a and its 

derivatives. Many other strains called T. 

reesei isolated elsewhere have now been 

proposed as belonging to a newly 

named species, T. parareesei, based on 

differences in habitat, sporulation, and 

metabolic versatility. T. reesei has been 

shown to belong to a single species now 

referred to as H. jecorina/T. reesei 

(QM6a) which reflects its relationship to 

its teleomorph H. jecorina. The only 

anamorphic strains within the species 

H. jecorina/T. reesei are those of QM6a 

and its derivatives. The petition to add 

T. reesei to the list of microorganisms at 

§ 725.420 requested that EPA include all 

strains of T. reesei. However, given 

these recent taxonomic publications, all 

fungal strains correctly named T. reesei 

are, by definition, QM6a or a derivative. 

Adequate genotypic and phenotypic 

information is available for 

classification of T. reesei QM6a and its 

derivatives. The American Type Culture 

Collection (ATCC) designation for this 

original strain of T. reesei QM6a is 

ATCC 13631. 

2. Information to evaluate the 

relationship of the microorganism to 

any other closely related 

microorganisms that have a potential 

for adverse effects on health or the 

environment. The petition to add T. 

reesei to the list of microorganisms at 

§ 725.420 requested that EPA include all 

strains of T. reesei. Closely related 

members of section Longibrachiatum do 

not have a potential for adverse effects; 

other less closely related Trichoderma 

species have a potential to cause 

adverse effects as pathogens of 

commercially produced mushrooms. 

These less closely related species 

include various species of the 

Harzianum clade, T. aggressivum, T. 

pleuotrophilum, and T. fulvidum that 

are responsible for significant loss of the 

mushroom crops of Agaricus bisporus 

and Pleurotus ostreatus. 

T. reesei/H. jecorina can be 

distinguished from other Trichoderma 

species by a comprehensive approach 

employing criteria of the Genealogical 

Concordance Phylogenetic Species 

Recognition (GCPSR) concept, which 

commonly requires the use of 

genealogies of three or four genes, not 

just the sequences of spacer regions as 

previously utilized for identification. 

Use of the GCPSR protocol will separate 

T. reesei (sensu lato) from the 

opportunistic pathogens within the 

section Longibrachiatum, including T. 

longibrachiatum and T. citronoviridae/ 

H. schweinitzii, as well as the mold 

disease pathogens of mushrooms. 

3. A history of safe commercial use for 

the microorganism. T. reesei QM6a has 

a long history of safe use producing a 

variety of commercial enzymes. T. 

reesei QM6a cellulases, beta-glucanases, 

and xylanases are used by the animal 

feed, baking, beverages, textile 

processing, detergent, pulp and paper, 

industrial chemicals, and biofuels 

industries. 

For industrial enzyme production, T. 

reesei is generally grown in a closed, 

submerged fermentation system. In 

submerged fermentation, growth of the 

microorganism occurs beneath the 

surface of the liquid growth medium. As 

described in this unit, this type of 

fermentation system appears to be 

typical throughout the industry, based 

on EPA’s review of MCAN submissions 

over the years. This type of fermentation 

system would also comply with the 

existing tiered exemption requirements 

relating to physical containment and 

control technologies, which are laid out 

in § 725.422. 

Under this type of fermentation 

system, the fermentation broth is a 

defined mixture of carbon and nitrogen 

sources, minerals, salts, and other 

nutrients, is maintained at optimal pH 

and temperature, and is typically 

aerated and mixed with no solid plant 

material or insoluble substrate present. 

These conditions support the active 

growth and productivity of the 

organisms. Submerged fermentation 

systems reduce the potential for 

exposure of workers to the production 

organism and fermentation broth 

aerosols, reduce the potential for 

contamination of the culture and make 

the collection of extracellular enzyme 

simpler and less costly. The 

fermentation process is terminated 

before the T. reesei QM6a organisms go 

into the stationary growth phase (i.e., 

before secondary metabolism begins). At 

the end of the fermentation process, the 

production organisms are separated 

from the fermentation broth and 

inactivated. Throughout the 

SUPPLEMENTARY INFORMATION section, 

EPA refers to this process as 

‘‘submerged standard industrial 

fermentation.’’ 

The Food and Drug Administration 

(FDA) has determined that several 

enzymes produced by T. reesei QM6a 

are Generally Recognized As Safe 

(GRAS). This determination supports 

the Agency’s preliminary conclusion 

that commercial use of T. reesei QM6a 

as a recipient microorganism for 

commercial enzyme production will not 

present an unreasonable risk of injury to 

health or the environment. T. reesei 

QM6a enzymes used in foods that have 

been granted GRAS status include 

cellulase, hemicellulase, 

transglucosidase, pectin lyase, acid 

fungal protease, and a chymosin enzyme 

preparation. Data supporting the GRAS 

petitions included the results of 

pathogenicity tests for the T. reesei 

QM6a production organisms and 

toxicity tests for the enzyme products. 

The data showed that the production 

strains are not pathogenic and did not 

produce toxins during enzyme 

fermentation. 

4. Commercial uses indicating that 

the microorganism products might be 

subject to TSCA. EPA has reviewed 

several MCANs involving intergeneric 

T. reesei QM6a production organisms. 

More detailed information on MCANs 

submitted to EPA can be viewed on 

EPA’s TSCA Biotechnology Program 

Web page: http://www.epa.gov/oppt/ 

biotech/pubs/submain.htm. 

Intergeneric T. reesei QM6a strains 

could also be used to manufacture 

industrial chemicals other than enzymes 

such as surfactants or specialty 

chemicals. 

5. Studies which indicate the 

potential for the microorganism to cause 

adverse effects to health or the 

environment—a. Human health 

hazards—i. Pathogenicity. Trichoderma 

reesei QM6a is not pathogenic to 

humans. Due to its long history of use 

for production of enzymes used in food 


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applications, the potential for the 

fungus and its products to be pathogenic 

or toxic to humans has been evaluated 

numerous times. Various studies have 

been conducted assessing T. reesei 

QM6a’s pathogenic potential in healthy 

and immunocompromised laboratory 

animals. Most studies have shown a 

lack of pathogenicity of T. reesei QM6a. 

Pathogenicity studies have been 

conducted as part of submissions 

submitted to FDA for GRAS petitions for 

several different enzymes used in the 

food industry. Studies using 

intraperitoneal (ip) injection of T. reesei 

QM6a in rats, using intravenous (IV) 

injection of T. reesei QM6a in both 

healthy and immunosuppressed rats, 

and using ip injection of viable and 

heat-killed cells of T. reesei QM6a in 

rats have all demonstrated a lack of 

potential pathogenicity to humans. 

T. reesei QM6A is not known to 

possess any virulence factors associated 

with colonization or disease such as 

adherence factors, penetration factors, 

necrotic factors, toxins, or the ability to 

grow at human body temperature, 37 °C. 

There are no reports in the literature on 

infection in healthy humans by T. reesei 

QM6A. There are no reports of harmful 

effects associated with the use of or 

exposure to T. reesei QM6A strains 

given decades of commercial use for 

enzyme production. The body of 

evidence indicates that T. reesei QM6A 

does not pose concerns regarding 

human pathogenicity. 

ii. Toxicity. Available data indicate 

that T. reesei QM6a strains used in 

submerged standard industrial 

fermentation operations in which no 

solid plant material or insoluble 

substrate is present in the fermentation 

broth do not present human toxicity 

concerns. A number of studies have 

been conducted assessing the potential 

for T. reesei QM6a to produce toxins 

during submerged fermentation for 

production of enzymes for food, 

pharmaceutical, or industrial uses. A 

cellulase enzyme known as celluclast 

produced by T. reesei QM6a has been 

tested for general oral toxicity and 

inhalation toxicity. Acute oral toxicity 

studies conducted in mice, rats, and 

dogs showed that T. reesei QM6a 

cellulase was not toxic to any of the test 

animals. Subchronic toxicity studies 

showed no evidence of systemic effects 

in dogs or rats. Additional toxicity 

studies have been conducted on other 

enzymes produced by T. reesei QM6a, 

the results of which have been 

presented in various GRAS petitions. 

Acute oral toxicity tests on two 

endoglucanases and a glucoamylase 

showed a lack of toxins. Subchronic 

feeding studies conducted on a 

cellulase, two xylanases, two 

endoglucanases, a protease, and a 

glucoamylase also showed a lack of 

toxicity in rats. 

Industrial strains of T. reesei QM6a 

are routinely checked by the enzyme 

producers to confirm the absence of 

antibiotic activity and toxins including 

aflatoxin B, ochratoxin A, 

sterigmatocystin, T–2 toxin, and 

zearalenone according to the 

recommendations of the Joint Food and 

Agriculture Organization and the World 

Health Organization (FAO/WHO) Expert 

Committee on Food Additives. Relying 

on the data that show T. reesei QM6a 

has a long history of safe use in the 

production of food enzymes where there 

is a need to routinely check for the 

absence of toxins, EPA has preliminarily 

concluded that strains used industrially 

would not be expected to produce these 

compounds under the growth 

conditions used for enzyme 

fermentation. 

iii. Mycotoxins and other secondary 

metabolites. The only health concern 

associated with T. reesei QM6a is its 

ability to produce a secondary 

metabolite called paracelsin, which is a 

peptaibol. Peptaibols are small linear 

peptides of 1,000–2,000 daltons 

characterized by a high content of the 

non-proteinogenic amino acid a-aminoisobutyric acid (Aib), with an Nterminus that is typically acetylated, 

and a C-terminus that is linked to an 

amino alcohol, which is usually 

phenylalaninol, or sometimes valinol, 

leucinol, isoleucinol, or tryptophanol. 

Peptaibols are associated with a wide 

variety of biological activities and have 

antifungal, antibacterial, sometimes 

antiviral, antiparasitic, and neurotoxic 

activity. Paracelsin has been shown to 

have toxicity toward mammalian cells 

such as hemolytic activity on human 

erythrocytes and cytotoxicity to rat 

adrenal medulla PC12 cells. Paracelsin 

showed toxicity to PC12 cells (a cell line 

derived from a pheochromocytoma of 

the rat adrenal medulla) with a CC50

(cytotoxicity concentration of 50%) of 

21.8 micromolar (mM) (Ref. 6). The in 

vitro hemolytic activity of paracelsin 

has been reported to be C50 = 3.7 × 10¥5

mole/liter (mol/L) (Ref. 6). 

Paracelsin has not been detected in 

the use of T. reesei QM6a under 

submerged standard industrial 

fermentation operations in which no 

solid plant material or insoluble 

substrate is present in the fermentation 

broth; numerous toxicity studies on 

enzyme products of T. reesei QM6a 

have demonstrated a lack of toxicity to 

laboratory animals. EPA therefore 

generally expects that paracelsin 

production will be of insignificant 

concern with submerged standard 

industrial fermentation operations in 

which no solid plant material or 

insoluble substrate is present in the 

fermentation broth. 

However, under non-standard 

conditions of fermentation, such as with 

extended duration of fermentation, or 

fermentation in the presence of 

insoluble carbon sources such as 

cellulose or in the presence of solid 

plant material, paracelsin may be 

produced (Ref. 6). Neither the 

information submitted with the petition, 

nor the information that is otherwise 

available is sufficient to allow EPA to 

determine the extent of paracelsin 

formation under these non-standard 

conditions. Consequently, EPA is 

unable to determine whether the use of 

the microbe under these non-standard 

conditions will pose an unreasonable 

risk to human health and/or the 

environment (Ref. 6). 

b. Environmental hazards—i. Hazards 

to animals. T. reesei QM6a is not 

pathogenic to domesticated animals or 

wildlife. However, the secondary 

metabolite paracelsin produced by T. 

reesei QM6a has been shown to exhibit 

toxicity to aquatic species. Twenty-four 

hour exposure of paracelsin to Artemia 

salina (brine shrimp) suggested a lethal 

concentration of 50% (LC50) of 21.26 mM 

(40.84 micrograms per milliliter (mg/ml)) 

which decreased to 9.66 mM (18.56 mg/ 

ml) with a 36-hour (hr) exposure. With 

Daphnia magna, paracelsin was found 

to be moderately toxic, with an LC50 of 

7.70 mM (14.79 mg/ml) with a 24-hr 

exposure, and 5.60 mM (10.76 mg/ml) 

with a 36-hr exposure. 

ii. Hazards to plants. Trichoderma 

reesei QM6a is not a pathogen of plants. 

Although it is capable of degrading 

cellulose and hemicellulose due to the 

copious quantities of the enzymes it can 

produce, it cannot be a primary 

colonizer on plant tissue as genetic 

studies have shown that it does not 

contain any genes for ligninases that are 

required for initial breakdown of plant 

material. This species is known as a 

wood rot fungus, but it apparently 

attacks only decaying plant material, not 

live plants. 

iii. Effects on other organisms. 

Peptaibols are toxic to Gram-positive 

bacteria and various fungi. The 

inhibitory action of peptaibols on 

various fungi is the reason that many 

species of Trichoderma are used as 

biocontrol agents of plant pathogenic 

fungi. T. reesei QM6a, which is known 

to produce only the peptaibol 

paracelsin, has been shown to be 

inhibitory to one particular fungus, 

Phoma destructiva. 


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Some species of Trichoderma, 

specifically T. aggressivum, T. 

pleuotrophilum, and T. fulvidum are 

pathogens of mushrooms. However, T. 

reesei QM6a is not a pathogen of 

mushrooms. 

6. Studies which indicate the survival 

characteristics of the microorganism in 

the environment. The species T. reesei 

is known only from the single original 

isolate QM6a from the Solomon Islands. 

Therefore, there is little information on 

its prevalence or behavior in the 

environment. Microcosm studies have 

been conducted that suggest it would 

survive in the environment if 

inadvertently released in the plant 

rhizosphere and in bulk soils. 

Although T. reesei was originally 

isolated from a tropical climatic region, 

it would be expected to persist in soils 

for extended periods of time, even after 

cold temperatures. 

B. Evaluation of Available Information 

Relevant to the Criteria at § 725.67 for 

B. amyloliquefaciens as a Recipient 

Microorganism 

1. Identification and classification of 

the microorganism using available 

genotypic and phenotypic information. 

Bacillus amyloliquefaciens was initially 

proposed as a unique species in 1943. 

The name Bacillus amyloliquefaciens 

lost standing when it was not included 

on the Approved List of Bacterial Names 

with Standing in Nomenclature in 1980. 

Since classical phenotypic tests could 

not differentiate it as a species unique 

from Bacillus subtilis, it was regarded as 

a subspecies of B. subtilis for several 

decades. However, molecular evidence 

from various subsequent studies led to 

the conclusion that Bacillus 

amyloliquefaciens did indeed deserve 

independent status. The DNA homology 

between B. subtilis and B. 

amyloliquefaciens is only about 15%. In 

addition, there were several phenotypic 

properties that differed between the two 

species. Chemotaxonomic studies 

revealed additional capability of 

separating strains of B. 

amyloliquefaciens from the other related 

species, B. subtilis, B. licheniformis, and 

B. pumilus. The species has remained 

within the genus Bacillus sensu stricto 

since it was last established as a 

separate species. 

Recently, it has been proposed that 

there are two subspecies within the 

species B. amyloliquefaciens, B. 

amyloliquefaciens subsp. 

amyloliquefaciens and B. 

amyloliquefaciens subsp. plantarum. 

The former subspecies includes the type 

strain and likely most, if not all, of the 

industrial strains of B. 

amyloliquefaciens used for enzyme 

production. The latter subspecies 

consists of plant-associated strains used 

as biocontrol agents since they produce 

a number of antifungal lipopeptide and 

antibacterial polyketide toxins. This 

proposed exemption would be restricted 

to the subspecies B. amyloliquefaciens 

subsp. amyloliquefaciens which 

contains the industrial strains used for 

enzyme production. Adequate genotypic 

and phenotypic information is available 

to accurately identify B. 

amyloliquefaciens subsp. 

amyloliquefaciens. 

2. Information to evaluate the 

relationship of the microorganism to 

any other closely related 

microorganisms which have a potential 

for adverse effects on health or the 

environment. There are several species 

in the genus Bacillus that are known 

pathogens. These include B. anthracis, 

which is pathogenic to humans and 

other animals, and B. cereus, which is 

a common cause of food poisoning. B. 

thuringiensis, B. larvae, B. lentimorbus, 

B. popilliae, and some strains of B. 

sphaericus are pathogenic or toxigenic 

to certain insects. The new subspecies 

B. amyloliquefaciens subsp. plantarum 

has been shown to exhibit toxicity 

mainly to plant pathogenic fungi, but 

can also be cytotoxic to mammalian 

cells. It is possible, using polyphasic 

approaches, to differentiate between 

Bacillus amyloliquefaciens subsp. 

amyloliquefaciens and these other 

species and subspecies that have the 

potential to adversely affect humans or 

other organisms. B. amyloliquefaciens 

can be distinguished from the very 

similar B. subtilis by a few phenotypic 

traits and DNA dissimilarity. 

3. A history of safe commercial use for 

the microorganism. Bacillus 

amyloliquefaciens has been used to 

produce commercial enzymes for more 

than 50 years. It produces 

carbohydrases, proteases, nucleases, 

xylanases, and phosphatases that have 

applications in the food, brewing, 

distilling, and textile industries. 

For commercial enzyme production, 

B. amyloliquefaciens is grown in a 

closed, submerged fermentation system. 

In submerged fermentation, growth of 

the microorganism occurs beneath the 

surface of the liquid growth medium. 

The fermentation broth is a defined 

liquid growth medium (with no solid 

plant material or insoluble substrate) of 

carbon and nitrogen sources, minerals, 

salts, and other nutrients that is 

maintained at optimal pH and 

temperature. These conditions support 

the active growth and productivity of 

the organisms. Submerged fermentation 

systems reduce the potential for 

exposure of workers to the production 

organism and fermentation broth 

aerosols, reduce the potential for 

contamination of the culture, and make 

the collection of extracellular enzyme 

simpler and less costly. The 

fermentation process is terminated 

before the B. amyloliquefaciens 

organisms go into the stationary growth 

phase (i.e., before secondary metabolism 

begins). At the end of the fermentation 

process, the production organisms are 

separated from the fermentation broth 

and inactivated. The enzyme 

preparation may also be subjected to 

other purification processes. 

B. amyloliquefaciens has a long 

history of safe use for the production of 

enzymes with both food and industrial 

uses with no incidences associated with 

human pathogenicity. In response to a 

petition from the ETA, FDA affirmed 

that carbohydrase enzyme preparations 

and protease enzyme preparations 

derived from either B. subtilis or B. 

amyloliquefaciens are GRAS for use as 

direct food ingredients. The European 

Food Safety Authority (EFSA) has put B. 

amyloliquefaciens on their list of 

bacteria that have a ‘‘qualified 

presumption of safety’’ (QPS) because of 

a long history of apparent safe use in 

food and feed production. However, it 

was put on the list with a qualifier that 

only strains of B. amyloliquefaciens that 

do not have toxigenic potential be used. 

One strain of B. amyloliquefaciens 

also has been used as a biopesticide. A 

naturally occurring strain of B. 

amyloliquefaciens subsp. plantarum 

was registered in 2000 as a biopesticide 

active ingredient under the Federal 

Insecticide, Fungicide, and Rodenticide 

Act (FIFRA). It can only be used on 

certain ornamental, non-food plants in 

greenhouses and other closed structures. 

4. Commercial uses indicating that 

the microorganism products might be 

subject to TSCA. It is expected that 

intergeneric strains of B. 

amyloliquefaciens will be used to 

produce enzymes and to manufacture 

other industrial chemicals subject to 

TSCA. Many enzymes produced by B. 

amyloliquefaciens, particularly aamylase, are used in laundry detergents 

and in textile processing. B. 

amyloliquefaciens also makes a 

surfactant known as surfactin which 

functions as an antibiotic. 

5. Studies which indicate the 

potential for the microorganism to cause 

adverse effects to health or the 

environment—a. Human health 

hazards—i. Pathogenicity. 

Bacillus amyloliquefaciens is not 

pathogenic to humans. There are no 

reports in the literature associating B. 

amyloliquefaciens with infection or 

disease in humans. B. amyloliquefaciens 


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has been categorized as a Biosafety 1 

microorganism by the Centers for 

Disease Control and Prevention (CDC). 

Biosafety 1 microorganisms are wellcharacterized agents not known to 

consistently cause disease in 

immunocompetent adult humans, and 

which present minimal potential hazard 

to laboratory personnel and the 

environment. Animal toxicity studies 

were performed with B. 

amyloliquefaciens strain FZB24 to 

support its registration as a biopesticide. 

Tests for acute oral toxicity/ 

pathogenicity, acute pulmonary 

toxicity/pathogenicity, and acute 

injection toxicity/pathogenicity showed 

little to no adverse effects, which 

indicated low mammalian toxicity and 

a lack of pathogenicity/infectivity. 

ii. Toxins and other secondary 

metabolites. Although another species 

in the genus Bacillus, B.cereus, has the 

potential to produce food poisoning 

toxins which cause both emetic and 

diarrheal syndromes, and a variety of 

local and systemic infections, the risk of 

food-borne disease caused by bacilli 

other than B. cereus is generally 

considered to be negligible because 

usually only B. cereus has the genes that 

encode food poisoning toxins. Industrial 

strains of Bacillus species belonging to 

the B. subtilis group, which includes B. 

amyloliquefaciens, do not express B. 

cereus toxins. In addition, there are no 

reported cases of food poisoning being 

caused by B. amyloliquefaciens. 

Some strains of B. amyloliquefaciens 

have been shown to produce bioactive 

cyclic lipopeptide metabolites such as 

iturin, surfactin, fengycin, and 

bacillomycin D. These are cyclical 

lipoprotein biosurfactants produced by 

non-ribosomal peptide synthesis. They 

have a low mammalian toxicity as 

demonstrated by a lethal dose of 50% 

(LD50) of >2,500 milligram/kilogram 

(mg/kg) in an acute toxicity test of 

surfactin C, and a No Observed Adverse 

Effect Level (NOAEL) of 500 mg/kg-day 

in a repeat dose oral gavage study. Some 

strains of B. amyloliquefaciens may also 

produce the polyketide toxins 

macrolactin, bacillanene, and difficidin. 

B. amyloliquefaciens also produces the 

protein toxin barnase and the antifungal 

protein baciamin. 

There are several reports of the 

isolation of B. amyloliquefaciens from 

water-damaged buildings in which 

occupants were suffering ill health 

symptoms. Extracts from biomass of 

isolated strains of Bacillus exhibiting 

antifungal properties were assessed for 

the toxicity endpoints. All of the 

isolated B. cereus and B. 

amyloliquefaciens strains studied 

showed cytotoxicity as evidenced by 

inhibition of boar spermatozoa motility; 

however, the B. amyloliquefaciens 

strains affected boar spermatozoa 

differently from the indoor B. cereus 

isolates and the reference foodpoisoning strain. 

The isolation of cytotoxic strains of B. 

amyloliquefaciens from water-damaged 

buildings is of little concern in relation 

to this exemption of B. 

amyloliquefaciens subsp. 

amyloliquefaciens. It is important to 

note that all of the B. amyloliquefaciens 

strains studied in water-damaged 

buildings were specifically selected for 

further study because the isolates 

exhibited antifungal activity. Some of 

the secondary metabolites produced by 

these biocontrol-type strains of B. 

amyloliquefaciens apparently also 

exhibit cytotoxicity to mammalian cells 

(i.e., boar spermatozoa). However, 

industrial strains of B. 

amyloliquefaciens that would fall into 

the classification as B. 

amyloliquefaciens subsp. 

amyloliquefaciens have been shown not 

to produce most, if not all, of the 

antifungal and antibacterial 

lipopeptides and polyketides produced 

by the biocontrol-type strains. The 

genome of the type strain of B. 

amyloliquefaciens DSM 7T (now B. 

amyloliquefaciens subsp. 

amyloliquefaciens) is very similar to the 

genome of the biocontrol strain FZB42 

(B. amyloliquefaciens subsp. 

plantarum). However, the latter 

subspecies had genomic islands 

carrying prophage sequences, 

transposases, integrases, and 

recombinases that the DSM 7T type 

strain did not have. The DSM 7T type 

strain was shown to have a diminished 

capacity to non-ribosomally synthesize 

secondary metabolites with antifungal 

and antibacterial activities. The DSM 7T

type strain could not produce the 

polyketides difficidin or macrolantin, 

and could not produce lipopeptide such 

as iturin, macrolantin, and other 

compounds except for the compound 

surfactin. 

The only other reported instance of 

mammalian toxin production by B. 

amyloliquefaciens was during the 1980s 

with the commercial production of 

tryptophan, by a genetically engineered 

strain of B. amyloliquefaciens, strain 

IAM 1521. The consumption of the 

tryptophan food supplement from 

various retail lots produced by one 

specific company resulted in an 

epidemic of a disease known as 

eosinophilia-myalgia syndrome (EMS) 

in which 1,511 were sickened, and 37 

people died. Although this disease 

incidence was widely studied, the cause 

of the disease was never confirmed. It 

was thought to be due to the 

consumption of a chemical constituent 

that was associated with specific 

tryptophan manufacturing processes. 

This included the combination of using 

reduced quantities of powdered carbon 

for a purification step with the use of a 

‘‘new’’ strain of B. amyloliquefaciens 

called Strain V. There purportedly was 

a chemical substance produced as a 

result of the genetic engineering of this 

certain strain, but the toxin was not 

attributable to the parental strain of B. 

amyloliquefaciens as not all production 

batches were toxic. 

Although there are isolated reports of 

toxin production in several antifungal, 

environmental isolates of B. 

amyloliquefaciens, the larger body of 

studies available on the safety and 

toxicity of B. amyloliquefaciens strains 

used industrially for enzyme production 

(Ref. 6) indicate that these strains are 

safe and non-toxic. For example, the 

toxicity of industrial strains of B. 

amyloliquefaciens, B. subtilis, and B. 

licheniformis used for large-scale 

enzyme production has been studied. 

The industrial strains did not exhibit 

any cytotoxicity in Chinese hamster 

ovary tests. In Europe, the toxicity of 

two strains of B. amyloliquefaciens used 

for the production of a-amylase and 

bacillolysin for the product Kemzyme W 

Dry was assessed by the EFSA’s 

Scientific Panel on Additives and 

Products or Substances used in Animal 

Feed. The panel concluded that the B. 

amyloliquefaciens production strains 

DSM9553 and DSM9554 when used as 

a source of extracellular enzyme do not 

present a toxigenic risk. Given its 

widespread distribution in the 

environment, its long history of safe use 

in industrial fermentation, the absence 

of reports on pathogenicity to humans, 

and the limited reports of cytotoxicity, 

all indicate that the use of B. 

amyloliquefaciens in fermentation 

facilities for production of enzymes or 

specialty chemicals does not present a 

human health concern. 

b. Environmental hazards—i. Hazards 

to animals. There are no reports 

suggesting that B. amyloliquefaciens is 

pathogenic to domesticated animals or 

wildlife. The cytotoxicity of antifungal 

secondary metabolites to mammalian 

cells by biocontrol stains of B. 

amyloliquefaciens is discussed in this 

unit. 

ii. Hazards to plants. B. 

amyloliquefaciens is not pathogenic to 

plants. There are plant-associated 

strains of B. amyloliquefaciens that are 

beneficial to plants because they inhibit 

the growth of fungal plant pathogens. 

Various antifungal and antibacterial 

secondary metabolites produced by 


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strains of B. amyloliquefaciens such as 

various iturins, surfactins, fengycin, 

bacillomycins, and azalomycin have 

been shown to inhibit the growth of 

Rhizoctonia solani, Xanthomonas 

campestris pv. campestris, Alternaria 

brassicae, Botyris cinerea, 

Leptosphaeria maculans, Verticillium 

longisporum, Pythium ultimatum, 

Aspergillus spp., Fusarium spp., 

Bipolaris sorokiniana, and Fusarium 

oxysporum. 

In addition to the ability of B. 

amyloliquefaciens to produce antifungal 

and antibacterial compounds, the 

bacterium is known as a plant growthpromoting rhizobacterium. Some of the 

biological control strains of B. 

amyloliquefaciens produce the 

phytohormone indole-3-acetic acid 

(IAA). 

6. Studies which indicate the survival 

characteristics of the microorganism in 

the environment. Using polymerase 

chain reaction (PCR) techniques, it has 

been found that populations of viable B. 

amyloliquefaciens inoculated at high 

densities to intact soil-core microcosms 

decreased to below the detection limit 

within 1 month. Survival was longer for 

a genetically modified B. 

amyloliquefaciens strain on leaf 

surfaces; vegetative cells were still 

detected for over 2 months in the 

phylloplane. Viable cells were not 

detectable in plant roots after 1 month 

or in soils after a few days. Given that 

the natural habitat for B. 

amyloliquefaciens is typically in soil, on 

plant roots, or as an endophyte within 

the roots or stems of plants, the 

bacterium is likely to survive for a least 

some period of time if inadvertently 

released to the environment. However, 

like other bacilli, survival in soil may 

occur predominately as the resistant 

endospore state, whereas in the 

rhizosphere, it may exist as active 

vegetative cells. 

IV. Physical Containment and Control 

Technologies 

A. Release and Exposure Assessment in 

Support of Proposed TSCA Section 

5(h)(4) Exemption for T. reesei QM6a 

The estimated releases of the 

microorganism from an enzyme 

manufacturing facility and exposures of 

the microorganisms to workers, the 

general population, and the 

environments are based on a generic 

scenario developed by EPA for largescale closed system fermentation. 

Assumptions in the generic scenario are 

that the facility operates 350 days/year, 

produces 100 batches/year, and the 

maximal cell concentration in the 

fermentation broth is 1 × 107 colonyforming units (cfu)/ml, and the volume 

of the fermentation broth is 70,000 L. 

The process consists of the main steps 

of laboratory propagation, fermentation 

and then recovery where filtration 

operations separate out the biomass 

from the concentrated desired product. 

The operations, sources of exposure and 

release are described in more detail in 

EPA’s Release and Exposure 

Assessments (Ref. 8). 

B. Release and Exposure Assessment in 

Support of Proposed TSCA 5(h)(4) 

Exemption for B. amyloliquefaciens 

The estimated releases of the 

microorganism from an enzyme 

manufacturing facility and exposures of 

the microorganisms to workers, the 

general population, and the 

environments are based on a generic 

scenario developed by EPA for largescale closed system fermentation. 

Assumptions in the generic scenario are 

that the facility operates 350 days/year, 

produces 100 batches/year, and the 

maximal cell concentration in the 

fermentation broth is 1 × 1011 cfu/ml 

and the volume of the fermentation 

broth is 70,000 L. The process consists 

of the main steps of laboratory 

propagation, fermentation and then 

recovery where filtration operations 

separate out the biomass from the 

concentrated desired product. The 

operations, sources of exposure and 

release are described in more detail in 

EPA’s Release and Exposure 

Assessments (Ref. 9). 

Additionally, containment and 

control technologies are delineated in 

the § 725.422 for Tier I and Tier II 

exemptions. 

V. Risk Assessment 

A. Risk Assessment for T. reesei QM6a 

There is only one potential concern 

for human health and environmental 

hazards associated with T. reesei QM6a, 

and that is for paracelsin production. 

Paracelsin production is not expected to 

occur in submerged standard industrial 

fermentation operations in which no 

solid plant material or insoluble 

substrate is present in the fermentation 

broth. There is no concern for potential 

pathogenicity of T. reesei QM6a to 

humans, plants, domesticated animals, 

or wildlife. Pathogenicity test data on 

various industrial strains typically do 

not show adverse effects. Toxicity 

testing on a number of enzymes 

produced by T. reesei indicates that the 

fungus does not produce toxins under 

the standard conditions used for 

enzyme production. 

T. reesei has a long history of safe use 

and would be expected to present low 

hazard to workers, the general public, 

and the environment. Although direct 

monitoring data are unavailable, worstcase estimates of potential exposures 

made by EPA in its assessment of 

potential risks (Ref. 6) do not indicate 

high levels of exposure of T. reesei to 

either workers or the public resulting 

from the submerged industrial enzyme 

fermentation operations that are 

standard throughout the industry. 

Standard industrial hygiene 

management practices currently used in 

the fermentation industry reduce the 

potential for adverse health effects in 

the workplace. The standard use of 

engineering controls (closed 

fermentation systems), appropriate work 

practices, personal protective 

equipment, and personal hygiene 

reduce the potential for worker 

exposure. Thus, current practices 

reduce the potential for the dermal and 

respiratory exposures estimated by EPA. 

EPA has made a preliminary 

determination based on worst-case 

exposure scenarios and toxicity of the 

microorganism that the potential risk to 

workers, the general public, and to the 

environment resulting from the use of T. 

reesei QM6a in submerged standard 

industrial fermentation as a recipient 

microorganism is low, provided the 

additional criteria of the tiered 

exemptions for the introduced genetic 

material and the physical containment 

conditions are met (Ref. 6). 

B. Risk Assessment for B. 

amyloliquefaciens 

Industrial strains of Bacillus 

amyloliquefaciens that would fall into 

the subspecies Bacillus 

amyloliquefaciens subsp. 

amyloliquefaciens are not pathogenic to 

humans, plants, domesticated animals, 

or wildlife, and do not produce many of 

the toxic secondary metabolites found 

in biological control strains of B. 

amyloliquefaciens subsp. plantarum. 

The long history of safe use of enzymes 

produced by industrial strains of B. 

amyloliquefaciens in food is evidence 

that the bacterium does not produce 

toxins under standard conditions used 

for enzyme production. 

Current practices in the fermentation 

industry reduce the potential for 

adverse health effects in the workplace. 

The use of engineering controls (closed 

fermentation systems), appropriate work 

practices, personal protective 

equipment, and personal hygiene 

reduce the potential for worker 

exposure. Thus, current practices 

reduce the potential for dermal and 

respiratory exposures. 

Industrial strains of B. 

amyloliquefaciens have a long history of 


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safe use and would be expected to 

present low hazard to workers, the 

general public, and the environment. 

Although direct monitoring data are 

unavailable, worst-case estimates do not 

suggest high levels of exposure of B. 

amyloliquefaciens to either workers or 

the public resulting from the submerged 

industrial enzyme fermentation 

operations that are standard throughout 

the industry. 

EPA has made a preliminary 

determination based on worst-case 

exposure scenarios and toxicity of the 

microorganism, that the potential risk to 

workers, the general public, and the 

environment, associated with the use of 

industrial strains of B. 

amyloliquefaciens subsp. 

amyloliquefaciens in submerged 

standard industrial fermentation as a 

recipient microorganism is low 

provided the additional criteria of the 

tiered exemptions for the introduced 

genetic material and the physical 

containment conditions are met (Ref. 7). 

VI. Economic Impacts 

EPA’s economic assessment (Ref. 10) 

evaluates the potential for significant 

economic impacts as a result of the 

addition of two microorganisms 

(Trichoderma reesei (Strain QM6a) and 

Bacillus amyloliquefaciens subsp. 

amyloliquefaciens) to § 725.420 which 

lists recipient microorganisms eligible 

for Tier I and Tier II exemptions. Over 

the course of the first 10 years after the 

effective date of the final rule, if 

finalized as proposed, EPA estimates 

that the proposed addition of the two 

microorganisms to the list in § 725.420 

would generate a total cost savings to 

society of $5.68 million. Industry would 

save approximately $1.98 million and 

the Agency would save approximately 

$3.68 million. The equivalent, 

annualized cost savings are expected to 

be $552,000 and $535,000 at a 3% and 

7% discount rate, respectively. EPA 

estimates that there will be a net 

decrease in burden to society of 72,500 

hr over this 10-year period. 

VII. Rationale for Proposed Regulatory 

Action 

A. Statutory Background 

Pursuant to TSCA section 5(h)(4), 

EPA is authorized to exempt the 

manufacturer of any new chemical 

substance from all or part of the 

requirements of TSCA section 5 if EPA 

determines that the manufacture, 

processing, distribution in commerce, 

use, or disposal of the chemical 

substance, or any combination of such 

activities, will not present an 

unreasonable risk of injury to human 

health or the environment. Section 26(c) 

of TSCA provides that any action 

authorized under TSCA for an 

individual chemical substance may be 

taken for a category of such chemical 

substances. 

While TSCA does not contain a 

definition of ‘‘unreasonable risk,’’ the 

legislative history indicates that the 

determination of unreasonable risk 

requires a balancing of the 

considerations of both the severity and 

the probability that harm will occur 

against the effect of the final regulatory 

action on the availability to society of 

the benefits of the chemical substance 

(Ref. 11). This analysis can include an 

estimate of factors such as market 

potential, the effect of the regulation on 

promoting or hindering the economic 

appeal of a chemical substance, 

environmental effects, and many other 

factors which are difficult to define and 

quantify precisely. EPA may rely not 

only on data available to it, but also on 

its professional judgment. Congress 

recognized that the implementation of 

the unreasonable risk standard ‘‘will 

vary on the specific regulatory authority 

which the Administrator seeks to 

exercise’’ [Ibid.]. 

B. EPA’s Approach 

In determining whether T. reesei 

QM6a and Bacillus amyloliquefaciens 

subsp. amyloliquefaciens will not 

present an unreasonable risk of injury to 

human health or the environment, the 

Agency considers more than just the 

inherent risks presented by the two 

microorganisms. The Agency also 

considers the full range of societal 

benefits associated with the exemption; 

for example, as discussed in more detail 

in Unit V., EPA considers not only the 

cost savings to the users of the 

microorganism, but also the societal 

benefits that flow from promotion of the 

use of low-risk recipient 

microorganisms, while allowing the 

Agency to direct its resources toward 

higher risk microorganisms. 

EPA is only proposing to revise one 

aspect of the existing tiered exemptions 

at § 725.420; specifically, EPA is 

proposing to expand the exemption to 

apply to two specific microorganisms. 

EPA is not reconsidering or otherwise 

reopening any other aspect of those 

exemptions. The narrow scope of this 

action necessarily affects the scope of 

EPA’s cost-benefit analysis. This means, 

for example, that EPA compares the 

risks and benefits of the two 

microorganisms being considered for an 

exemption with the risks that would 

have resulted if those same two 

microorganisms remained subject to full 

MCAN submission requirements and 

90-day EPA review. But EPA does not 

compare the risks and benefits that 

would result from use of these two 

microorganisms in the absence of any 

regulation. 

It is also significant that the standard 

applicable to this proposed rule is that 

the microorganisms will present ‘‘no 

unreasonable risk,’’ rather than ‘‘no 

risk.’’ It is not possible to eliminate all 

risks associated with the manufacture, 

processing, distribution in commerce, 

use, and disposal of any new 

microorganism nor was this Congress’ 

intent. The standard embodied by a 

TSCA section 5(h)(4) exemption does 

not require the Agency to ensure 

absolute safety from the activities 

associated with an exempted chemical 

substance. 

C. Application of No Unreasonable Risk 

Factors 

The following is an explanation of the 

factors and their analyses relevant to the 

no unreasonable risk finding. 

1. Risks associated with 

microorganisms. EPA’s evaluation of the 

available information concerning T. 

reesei QM6a and B. amyloliquefaciens 

subsp. amyloliquefaciens against these 

criteria is presented in detail in Unit III., 

and is summarized again here for the 

readers’ convenience. 

The Agency developed specific 

criteria in § 725.67 that the Agency uses 

in determining the extent of a potential 

recipient microorganism’s risks, and 

consequently, its eligibility for listing at 

§ 725.420. These criteria were explained 

in detail in the proposed ‘‘biotech’’ rule 

(Ref. 12) and final ‘‘biotech’’ rule (Ref. 

13), and are discussed again in Units II. 

and III. EPA’s conclusions regarding the 

low-risk potential for these two 

microorganisms are based on the 

available data and EPA’s scientific 

professional judgment based on 14 years 

experience reviewing notifications for 

new intergeneric microorganisms 

submitted in accordance with the 

regulations at 40 CFR part 725. 

T. reesei QM6a is not pathogenic to 

humans, plants, domesticated animals, 

or wildlife and the fungus does not 

produce toxins under standard 

industrial conditions used for enzyme 

production. T. reesei QM6a has a long 

history of safe use and is generally 

expected to present low risk to workers, 

the general public, and the environment 

resulting from submerged standard 

industrial enzyme fermentation 

operations that are standard throughout 

the industry. Under non-standard 

conditions of fermentation, such as with 

extended duration of fermentation, or 

fermentation in the presence of 

insoluble carbon sources such as 


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cellulose or other solid surfaces, 

paracelsin may be produced. The risks 

associated with the production of 

paracelsin may be significant due to the 

toxicity of paracelsin to mammalian 

cells, aquatic species, Gram-positive 

bacteria, and various fungi. However, 

the potential risk associated with any 

paracelsin production would be 

significantly reduced by this proposed 

rule, which proposes to limit the 

exemption to fermentation operations 

using submerged standard industrial 

fermentation operations, and in which 

no solid plant material or insoluble 

substrate is present in the fermentation 

broth. 

Industrial strains of Bacillus 

amyloliquefaciens that would fall into 

the subspecies Bacillus 

amyloliquefaciens subsp. 

amyloliquefaciens are not pathogenic to 

humans, plants, domesticated animals, 

or wildlife, and do not produce toxins 

under standard conditions used for 

enzyme production. Industrial strains of 

B. amyloliquefaciens subsp. 

amyloliquefaciens used in fermentation 

facilities for the production of enzymes 

have a long history of safe use and are 

expected to present low hazards to 

human health and the environment 

resulting from standard industrial 

submerged fermentation operations. 

Consistent with the proposed 

restrictions on Trichoderma reesei 

discussed in Unit II.A., only strains of 

Bacillus amyloliquefaciens that would 

fall into the subspecies Bacillus 

amyloliquefaciens subsp. 

amyloliquefaciens were considered as 

the eligible recipient microorganism at 

§ 725.420. EPA is proposing to exclude 

other strains/subspecies of these two 

species for which: 

• The Agency still has insufficient 

data and review experience to find that 

they will not present an unreasonable 

risk of injury or 

• The Agency has found that, under 

certain conditions, based on data on the 

species in question, a strain or 

subspecies may present an unreasonable 

risk, thereby requiring a closer 

examination of the conditions of 

manufacturing, processing, distribution 

in commerce, use, and disposal during 

a full 90-day Premanufacture Notice 

(PMN) review. Consequently, additional 

information would be necessary to make 

an appropriate determination about the 

organisms’ potential risks and benefits. 

The Agency believes that the 

requirement for submission of a MCAN 

followed by a 90-day review period for 

new intergeneric microorganisms that 

use T. reesei QM6a and Bacillus 

amyloliquefaciens subsp. 

amyloliquefaciens as recipient 

microorganisms is not necessary to 

address the risks associated with these 

microorganisms, and would not result 

in any additional protection than would 

be achieved by this proposed rule. In 

part, this conclusion is based on EPA’s 

preliminary findings regarding the 

intrinsically low level of hazard that 

these two organisms pose to human 

health and the environment. In 

addition, the existing requirements of 

the Tier I and Tier II exemptions, taken 

with the proposed restrictions, would 

place sufficient constraints to 

significantly limit the potential risks of 

injury to human health or the 

environment that these two 

microorganisms may present. 

In sum, the Agency believes that the 

criteria set forth in this proposed 

exemption would be sufficient to 

mitigate the identified risks associated 

with these microorganisms. 

2. Costs. This proposed rule expands 

an existing exemption, and as discussed 

in Unit VI., would significantly reduce 

costs to currently regulated entities. The 

proposed rule would not otherwise 

impose any additional cost or other 

burden on currently regulated entities, 

or existing fermentation processes. 

EPA further believes that limiting the 

use of this proposed exemption to the 

identified fermentation conditions 

would impose no burden on affected 

entities. The restriction merely codifies 

existing industrial fermentation 

procedures that are common practices 

for manufacturing operations that 

currently seek to use tiered exemptions. 

Consequently, EPA expects that most, if 

not all, manufacturers currently using 

these microbes will already have the 

measures in place to qualify for the 

exemption. Equally important, this 

limitation would add no burden to any 

existing fermentation processes. 

Currently, fermentation operations with 

either of these microbes are not eligible 

for the tiered exemption, and thus a 

MCAN must be submitted. Any 

company that chooses to use a different 

fermentation process could continue to 

operate under the status quo and simply 

submit a MCAN. This proposed rule 

would simply offer an additional, less 

costly option, to facilities that choose to 

use the fermentation operations 

discussed in this proposed rule. 

3. Benefits. The following discussion 

describes the benefits of this proposed 

rule in a qualitative manner; for a more 

quantitative approach, see the economic 

analysis prepared for this proposed rule 

(Ref. 10). A summary of that economic 

analysis is also provided in Unit VI. 

The benefits analyzed encompass 

more than the direct benefits associated 

with submitting a Tier I or Tier II 

exemption for a new intergeneric 

microorganism rather than a MCAN. 

Rather, EPA’s benefit analysis included 

a consideration of the broader benefits 

to society. EPA’s unreasonable risk 

determination is based on broader 

benefits to society as well as those 

benefits attributable to a reduction in 

the burden associated with submission 

of Tier I and Tier II exemptions rather 

than MCANs. 

EPA believes manufacturers of new 

intergeneric microorganisms based on 

these low-risk microorganisms currently 

bear an unnecessary regulatory burden 

in continuing to file MCANs. By adding 

T. reesei QM6a and B. 

amyloliquefaciens to the list of eligible 

recipient microorganisms in § 725.420, 

the Agency removes unnecessary 

regulatory impediments to the design, 

manufacture, and commercialization of 

these low risk new intergeneric 

microorganisms, and of the chemical 

substances that can be produced by 

these safer microorganisms. This action 

would also substantially reduce the 

costs associated with industry’s 

reporting burden, including the costs 

associated with the preparation of the 

submission, and with the delay in the 

commercial market introduction of the 

new intergeneric microorganism. Some 

of the cost-savings benefits may accrue 

to small businesses, either as developers 

of the exempt microorganisms, as 

producers of fermentation chemicals 

using the live microorganisms, or as 

customers for enzymes or other 

products made using the 

microorganisms. 

There would also be a reduction in 

the Agency review resources currently 

allocated to reviews of MCANs for these 

two microorganisms. These Agency 

resources would be shifted to the review 

of new intergeneric microorganisms or 

chemical substances of greater concern. 

There would be cost savings to both 

the industry and the Agency. The 

proposed rule is expected to positively 

impact the rate of innovation in the 

industry. It is reasonable to assume that 

a new intergeneric microorganism will 

either possess a new function or serve 

an existing function more efficiently or 

less expensively. The reduction in delay 

for that new intergeneric microorganism 

to be introduced into commerce is a 

benefit to both manufacturers and the 

general public who will have access to 

the substance more quickly. The 

expected benefits to innovation have not 

been quantified but include: Reduced 

time to develop and commercialize 

organisms; decreased cost of some 

downstream industrial products, such 

as fuel ethanol; improved consumer 

appeal of some products, such as certain 


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textiles; and reduced costs of some 

consumer products, such as detergent 

and leather goods. 

4. Risk/benefit balance. Determining 

the presence or absence of an 

unreasonable risk requires balancing of 

the benefits and risks posed by a 

regulatory action. EPA has determined 

that the risks are generally low based on 

the inherent properties and intended 

uses of T. reesei QM6a and B. 

amyloliquefaciens, and would be 

adequately managed by the restrictions 

in the proposed rule, combined with the 

existing requirements of the Tier I and 

Tier II exemptions. 

As noted in this unit, EPA believes 

that this proposed rule would impose 

no costs. This proposed rule expands an 

existing exemption, and as such, would 

in fact reduce costs to currently 

regulated entities. This proposed rule 

would not otherwise impose any 

additional cost or other burden on 

currently regulated entities, or existing 

fermentation processes. The limitation 

on the use of the proposed exemption to 

certain fermentation conditions is not a 

cost that would be imposed by this 

proposed rule but rather a limitation on 

the amount of regulatory relief it would 

provide. The proposed conditions 

reflect industrial fermentation 

procedures that are currently common 

practices for the affected industry. 

EPA also believes that the benefits of 

this proposed rule are quite significant. 

This proposed rule would reduce the 

overall regulatory burden for affected 

entities by reducing the reporting 

requirements and by eliminating the 

delay of these products into commerce. 

As a consequence, this would benefit 

both regulated entities and the general 

public by promoting the expedited 

manufacture and use of the chemical 

substances produced using these lowrisk organisms and manufacturing 

processes. There is also the added 

benefit of concentrating limited EPA 

resources on regulation of chemical 

substances which have a greater 

potential to present significant risks, 

rather than on these two 

microorganisms. While this is difficult 

to quantify, it is considered substantial 

nonetheless. 

In sum, the Agency believes that the 

criteria set forth in this proposed 

exemption are sufficient to mitigate the 

low level of potential risks presented by 

these organisms, particularly when 

compared to the benefits, in toto, of this 

proposed exemption, to levels that are 

consistent with the statutory standard 

for an exemption. Consequently, EPA 

has made a preliminary conclusion that 

adding T. reesei QM6a and B. 

amyloliquefaciens as recipient 

microorganisms to the list of recipient 

microorganisms at § 725.420 is 

appropriate, as it would not present an 

unreasonable risk of injury to human 

health or the environment when 

manufactured under the conditions of 

this proposed exemption. 

VIII. Request for Public Comment, 

Rulemaking Process, and Request for 

an Informal Public Hearing 

A. Rulemaking Process and Request for 

an Informal Public Hearing 

EPA is conducting this rulemaking 

under the notice and comment 

rulemaking procedures of section 553 of 

the Administrative Procedure Act 

(APA), 5 U.S.C. 553. Interested persons 

have the opportunity to submit written 

comments by the methods identified 

under ADDRESSES. EPA will carefully 

consider all such comments. 

EPA is also providing an opportunity 

for an informal public hearing on the 

proposed rule. This hearing will be held 

only if EPA receives a timely written 

request for such a hearing. 

As a general matter, EPA is not 

required to hold a public hearing in 

informal notice and comment 

rulemaking conducted under APA 

section 553. However, use of TSCA 

section 5(h)(4) modifies the APA section 

553 rulemaking requirements by 

referencing TSCA section 6(c)(2) and 

(c)(3) rulemaking procedures. Under the 

TSCA section 6 procedures, EPA must 

hold an informal public hearing, if 

requested, and, if properly requested 

and granted by EPA, allow an 

opportunity to present rebuttal 

submissions and conduct crossexaminations related to disputed issues 

of material fact. 

EPA does not anticipate that, even if 

a hearing is held, there will be a need 

for rebuttal submissions and crossexamination, because the TSCA section 

5(h)(4) portion of this proposed 

rulemaking is based primarily on 

matters of science policy that do not 

yield disputed factual issues. 

B. Specific Comment Solicitation 

EPA is seeking public comment 

pertaining to several specific issues 

regarding the proposed rule. 

1. Do the proposed rule and 

supporting documents adequately 

address: 

• The effects of the new 

microorganism on health and the 

environment? 

• The magnitude of exposure of 

human beings and the environment to 

the new microorganism? 

• The benefits of the new 

microorganism for various uses and the 

availability of substitutes for such uses? 

• The reasonably ascertainable 

economic consequences of granting or 

denying the exemption, including 

effects on the national economy, small 

business, and technological innovation? 

2. Does the proposed rule address 

taxonomy adequately (is the Agency 

capturing and excluding the correct 

strains)? 

3. Does the proposed rule address the 

right description of typical conditions 

for enzyme production (eliminating 

plant material/solid surfaces)? 

4. Are the limitations on the use of T. 

reesei QM6a reasonable for preventing 

paracelsin production (i.e., having no 

solid plant material or insoluble 

substrate with the microorganism)? 

IX. References 

As indicated under ADDRESSES, a 

docket has been established for this 

proposed rule under docket ID number 

EPA–HQ–OPPT–2011–0740. The 

following is a listing of the documents 

that have been placed in the docket for 

this proposed rule. The docket includes 

information considered by EPA in 

developing this proposed rule, 

including the documents listed in this 

unit, which are physically located in the 

docket. In addition, interested parties 

should consult documents that are 

referenced in the documents that EPA 

has placed in the docket, regardless of 

whether these referenced documents are 

physically located in the docket. For 

assistance in locating documents that 

are referenced in documents that EPA 

has placed in the docket, but that are 

not physically located in the docket, 

please consult the technical contact 

listed under FOR FURTHER INFORMATION

CONTACT. The docket is available for 

review as specified under ADDRESSES. 

1. Genencor International, Inc. Letter of 

Application to list Trichoderma reesei as 

exempt under subpart G of 40 CFR Part 725— 

Reporting Requirements and Review 

Processes for Microorganisms. March 17, 

2005. 

2. Novo Nordisk BioChem North America, 

Inc. Letter of Application to list 

B.amyloliquefaciens as exempt under subpart 

G of 40 CFR Part 725—Reporting 

Requirements and Review Processes for 

Microorganisms. November 7, 1997. 

3. EPA, OPPT. Email confirming Novo 

Nordisk BioChem North America, Inc.’s letter 

of application to list B.amyloliquefaciens as 

exempt under subpart G of 40 CFR Part 725— 

Reporting Requirements and Review 

Processes for Microorganisms. August 3, 

2009. 

4. ETA. Supplemental information on 

Trichoderma reesei. January 29, 2010. 

5. ETA. Supplemental information on 

Trichoderma reesei. June 16, 2011. 

6. EPA, OPPT. Risk Assessment of 

Trichoderma reesei for Consideration of 

Addition to the List of Eligible Recipient 


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Microorganisms for the Tiered 5(h)(4) 

Exemptions from MCAN Reporting 

Requirements. October 2011. 

7. EPA, OPPT. Risk Assessment of Bacillus 

amyloliquefaciens subsp. amyloliquefaciens 

for Consideration of Addition to the List of 

Eligible Recipient Microorganisms for the 

Tiered 5(h)(4) Exemptions from MCAN 

Reporting Requirements. October 2011. 

8. EPA, OPPT. Release and Exposure 

Assessment in Support of Proposed TSCA 

5(h)(4) Exemption for Trichoderma reesei. 

June 2011. 

9. EPA, OPPT. Release and Exposure 

Assessment in Support of Proposed TSCA 

5(h)(4) Exemption for Bacillus 

amyloliquefaciens. June 2011. 

10. EPA, OPPT. Economic Analysis for the 

Proposed Biotechnology Exemptions Rule for 

Trichoderma reesei and Bacillus 

amyloliquefaciens. September 2011. 

11. Legislative History of the Toxic 

Substances Control Act, pp. 409–423. House 

Report 1341, 94th Congress, 2nd Session. 

1976. 

12. EPA. Microbial Products of 

Biotechnology; Proposed Regulation under 

the Toxic Substances Control Act. Federal 

Register (59 FR 45526; September 1, 1994) 

(FRL–4774–4). 

13. EPA. Microbial Products of 

Biotechnology; Final Regulation under the 

Toxic Substances Control Act. Federal 

Register (62 FR 17910; April 11, 1997) (FRL– 

5577–2). 

X. Statutory and Executive Order 

Reviews 

A. Executive Order 12866: Regulatory 

Planning and Review and Executive 

Order 13563: 

Improving Regulation and Regulatory 

Review 

This action is not a ‘‘significant 

regulatory action’’ under the terms of 

Executive Order 12866 (58 FR 51735, 

October 4, 1993) and is therefore not 

subject to review under Executive 

Orders 12866 and 13563 (76 FR 3821, 

January 21, 2011). EPA prepared an 

analysis of the potential costs and 

benefits associated with this action, 

which is summarized in Unit VI. 

B. Paperwork Reduction Act (PRA) 

According to PRA, 44 U.S.C. 3501 et 

seq., an agency may not conduct or 

sponsor, and a person is not required to 

respond to, a collection of information 

that requires approval by the Office of 

Management and Budget (OMB) under 

PRA, unless it has been approved by 

OMB and displays a valid OMB control 

number. The OMB control numbers for 

EPA’s regulations in title 40 of the CFR, 

after appearing in the Federal Register, 

are listed in 40 CFR part 9, and included 

on the related collection instrument, or 

form, if applicable. 

The information collection 

requirements related to the submission 

of Tier I and Tier II notification are 

already approved by OMB under PRA, 

and have been assigned OMB control 

numbers 2070–0012 and 2070–0038. 

This proposed rule does not impose any 

new requirements, or otherwise increase 

burden such that additional OMB 

review or approval is necessary. Instead, 

this proposed rule is expected to reduce 

the amount of required reporting by 

allowing firms to submit less 

information for qualifying 

microorganisms. 

The PRA requires agencies to estimate 

the potential recordkeeping and 

reporting burden of a proposed rule. In 

this context, the term ‘‘burden’’ is 

defined in 5 CFR 1320.3(b). EPA 

estimates that this proposed rule would 

result in a reduction of industry burden 

by 30,695 hr over 10 years. EPA also 

estimates that the proposed rule would 

cause a total incremental Agency 

savings of 41,869 hr over 10 years. 

Submit any comments related to these 

estimates to EPA. See ADDRESSES for 

submission of comments. 

C. Regulatory Flexibility Act (RFA) 

Pursuant to section 605(b) of the RFA, 

5 U.S.C. 601 et seq., the Agency hereby 

certifies that this proposed rule, if 

promulgated as proposed, would not 

have a significant economic impact on 

a substantial number of small entities. 

Under RFA, small entities include small 

businesses, small organizations, and 

small governmental jurisdictions. For 

purposes of assessing the impacts of this 

action on small entities, small entity is 

defined as: 

1. A small business as defined by the 

Small Business Administration’s (SBA) 

regulations at 13 CFR 121.201 using 

either the number of employees or 

annual receipts for the businesses 

affected by the regulation, which for this 

action includes any business that is 

conducting commercial research and 

development activities or persons 

manufacturing, importing or processing 

products using intergeneric 

microorganisms for biofertilizers; 

biosensors; enzyme, commodity, or 

specialty chemical production; energy 

applications; waste treatment or 

pollutant degradation; and other TSCA 

subject uses. 

2. A small governmental jurisdiction 

that is a government of a city, county, 

town, school district, or special district 

with a population of less than 50,000. 

3. A small organization that is any 

not-for-profit enterprise which is 

independently owned and operated and 

is not dominant in its field. 

In making this determination, the 

impact of concern is any significant 

adverse economic impact on small 

entities because the primary purpose of 

regulatory flexibility analysis is to 

identify and address regulatory 

alternatives ‘‘which minimize any 

significant economic impact of the rule 

on small entities.’’ 5 U.S.C. 603 and 604. 

Thus, an agency may certify under RFA 

when the rule relieves regulatory 

burden, or otherwise has no expected 

economic impact on small entities 

subject to the rule. 

This proposed rule is an exemption, 

and is therefore expected to reduce the 

existing regulatory burden, which will 

benefit all submitters regardless of the 

size of the entity. The factual basis for 

the Agency’s certification under RFA is 

presented in the small entity impact 

analysis prepared as part of the 

Economic Analysis for this proposed 

rule (Ref. 10), and is briefly summarized 

in Unit VI. 

We continue to be interested in the 

potential impacts of the proposed rule 

on small entities and welcome 

comments on issues related to such 

impacts. 

D. Unfunded Mandates Reform Act 

(UMRA) 

EPA has determined that this action 

does not impose any enforceable duty or 

contain any unfunded mandate for 

State, local, or Tribal governments or 

the private sector, and does not 

otherwise have any effect on small 

governments, such that it is subject to 

the requirements of sections 202, 203, 

204, or 205 of UMRA, 2 U.S.C. 1531– 

1538. As indicated previously, this 

action is expected to reduce costs. In 

addition, based on EPA’s experience 

with past MCANs and Tier I and II 

exemptions, State, local, and Tribal 

governments have not been affected by 

these reporting requirements, and EPA 

does not have any reason to believe that 

any State, local, or Tribal government 

will be affected by this particular 

rulemaking. A search of past 

submissions to EPA demonstrated that 

no State, local, or Tribal government 

have ever submitted a MCAN, Tier I or 

Tier II notification to EPA. EPA has no 

information to indicate that any State, 

local, or Tribal government 

commercially manufactures the 

microorganisms covered by this action. 

E. Executive Order 13132: Federalism 

For the same reasons presented in 

Unit X.D., the Agency has determined 

that this action will not have a 

substantial direct effect on State or local 

governments, on the relationship 

between the national government and 

the States or local governments, or on 

the distribution of power and 

responsibilities among the various 


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levels of government. Thus, the Agency 

has determined that Executive Order 

13132 (64 FR 43255, August 10, 1999) 

does not apply to this action. 

F. Executive Order 13175: Consultation 

and Coordination With Indian Tribal 

Governments 

For the same reasons presented in 

Unit X.D., the Agency has determined 

that this action will not have a 

substantial direct effect on tribal 

governments, on the relationship 

between the national government and 

Tribal governments, or on the 

distribution of power and 

responsibilities between the Federal 

Government and Indian tribes. Thus, the 

Agency has determined that Executive 

Order 13175 (65 FR 67249, November 9, 

2000) does not apply to this action. 

G. Executive Order 13045: Protection of 

Children From Environmental Health 

Risks and Safety Risks 

EPA interprets Executive Order 13045 

(62 FR 19885, April 23, 1997) as 

applying only to those regulatory 

actions that concern health or safety 

risks, such that the analysis required 

under section 5–501 of the Executive 

Order has the potential to influence the 

regulation. This action is not subject to 

Executive Order 13045 because it does 

not establish an environmental standard 

intended to mitigate health or safety 

risks, nor is it an ‘‘economically 

significant regulatory action’’ as defined 

by Executive Order 12866. 

H. Executive Order 13211: Actions 

Concerning Regulations That 

Significantly Affect Energy Supply, 

Distribution, or Use 

This action is not subject to Executive 

Order 13211 (66 FR 28355, May 22, 

2001), because it is not a significant 

regulatory action under Executive Order 

12866. 

I. National Technology Transfer and 

Advancement Act (NTTAA) 

Section 12(d) of NTTAA, 15 U.S.C. 

272 note, directs EPA to use voluntary 

consensus standards in its regulatory 

activities unless to do so would be 

inconsistent with applicable law or 

impractical. Voluntary consensus 

standards are technical standards (e.g., 

materials specifications, test methods, 

sampling procedures, etc.) that are 

developed or adopted by voluntary 

consensus standards bodies. This 

proposed rule does not impose any 

technical standards that would require 

EPA to consider any voluntary 

consensus standards. 

J. Executive Order 12898: Federal 

Actions To Address Environmental 

Justice in Minority Populations and 

Low-Income Populations 

This action does not have 

disproportionately high and adverse 

human health or environmental effects 

on minority or low-income populations 

because it does not affect the level of 

protection provided to human health or 

the environment. Therefore, this action 

does not involve special consideration 

of environmental justice-related issues 

as specified in Executive Order 12898 

(59 FR 7629, February 16, 1994). 

List of Subjects in 40 CFR Part 725 

Environmental protection, 

Administrative practice and procedure, 

Biotechnology, Chemicals, Hazardous 

substances, Imports, Labeling, 

Microorganisms, Occupational safety 

and health, Reporting and 

recordkeeping requirements. 

Dated: August 28, 2012. 

James Jones, 

Acting Assistant Administrator, Office of 

Chemical Safety and Pollution Prevention. 

Therefore, it is proposed that 40 CFR 

chapter I be amended as follows: 

PART 725—[AMENDED] 

1. The authority citation for part 725 

continues to read as follows: 

Authority: 15 U.S.C. 2604, 2607, 2613, and 

2625. 

2. In § 725.3, add in alphabetical order 

the definition below to read as follows: 

§ 725.3 Definitions. 

* * * * * 

Submerged standard industrial 

fermentation for purposes of this part, 

means a fermentation system that meets 

all of the following conditions: 

(1) Submerged fermentation (i.e., 

growth of the microorganism occurs 

beneath the surface of the liquid growth 

medium). 

(2) Any fermentation of solid plant 

material or insoluble substrate, to which 

T. reesei fermentation broth is added 

after the standard industrial 

fermentation is completed, may be 

initiated only after the inactivation of 

the microorganism as delineated in 

§ 725.422(d). 

* * * * * 

3. In § 725.420, add new paragraphs 

(k) and (l) to read as follows: 

§ 725.420 Recipient microorganisms. 

* * * * * 

(k) Trichoderma reesei strain QM6a 

used only in submerged standard 

industrial fermentation operations in 

which no solid plant material or 

insoluble substrate is present in the 

fermentation broth, fermentation may 

only be initiated after the inactivation of 

T. reesei as delineated in § 725.422(d). 

(l) Bacillus amyloliquefaciens subsp. 

amyloliquefaciens. 

[FR Doc. 2012–21843 Filed 9–4–12; 8:45 am] 

BILLING CODE 6560–50–P 

FEDERAL COMMUNICATIONS 

COMMISSION 

47 CFR Part 101 

[WT Docket No. 10–153; FCC 12–87] 

Facilitating the Use of Microwave for 

Wireless Backhaul and Other Uses and 

Providing Additional Flexibility To 

Broadcast Auxiliary Service and 

Operational Fixed Microwave 

Licensees 

AGENCY: Federal Communications 

Commission. 

ACTION: Proposed rule. 

SUMMARY: In this document, the 

Commission seeks more detailed 

comments on specific proposals made 

by parties to allow use of smaller 

antennas and wider channels in other 

part 101 microwave bands. We also seek 

comment on a proposal to revise our 

rules to change our treatment of smaller 

antennas in the 10.7–11.7 GHz band (11 

GHz band). We also seek comment on 

additional ways to increase the 

flexibility, capacity, and costeffectiveness of the microwave bands, 

while protecting incumbent licensees in 

these bands. In the Second Notice of 

Inquiry, we seek comment on making 

additional changes to our antenna 

standards to reflect advances in 

technology, accommodate non-parabolic 

antennas, and harmonize our standards 

with international standards. By 

enabling more flexible and cost-effective 

microwave services, the Commission 

can help foster deployment of 

broadband infrastructure across 

America. 

DATES: Submit comments on or before 

October 5, 2012. Submit reply 

comments on or before October 22, 

2012. 

ADDRESSES: Federal Communications 

Commission, 445 12th Street SW., 

Washington, DC 20554. You may submit 

comments, identified by FCC 12–87, or 

by WT Docket No. 10–153, or by any of 

the following methods: 

Federal eRulemaking Portal: http:// 

www.regulations.gov. Follow the 

instructions for submitting comments. 

Federal Communications 

Commission’s Web Site: http:// 


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