Marine Mammal Health and 

Stranding Response I'rogram: 

Program Devellopment Plan 

Prepared by 

Paul R. Becker, Dean Wilkinson, and Ted I. Lillestolen 

National Marine Fisheries Service 

Office of Protected Resources 

July 1994 

U.S. Department Of Comnnerce 

National Oceanic and Atmospheric A dminis tration~ 

Na tional Marine Fisheries Se~wict? 
Marine Mammal Health and 

Stranding Response Program: 

Program Development Plan 

Paul I<. Beclter, Dean Wilkinson, and 

Ted I. Lillestolen 

National Marine Fisheries Service 

Office of Protected Resources 

NOA,A Technical Memorandum 

NMFS-OPIY-94-2 

July 1994 

U.S. Department of Commerce 

Ronald H. Brown, Secretary $+'*' O' c, 

s '%; National Oceanic and Atmospheric Administration 

D. James Baker, Under Secretary for Oceans and A,tm~os;phere 

C 

'~*~~50vr*' National Marine Fisheries Service 

Rolland! A. Schn~itten, Assistant Administrator for Fisheries 
Abstract 

The Marine Mammal Health and Stranding Response Program (Mh/IHSR.P) was estatbl~ished under 

the authority of Title I11 of the Marine Mammal Protection Act to faci1itat.e the collection and 

di:;semination of reference data on health trends in marine mammal populations, correl,ate the 

health of marine mammals with environmental parameters, and cc~ordintate c:ffective r1espna.s to 

un,usual mortality events. The components of the Program are: the National Marine Marlnmal Tissue 

Bank (NMMTB), the Stranding Network, Monitoring, and Quality Assurance. The N'MMTB was 

established to provide a resource of research materials for future reb:ospective i3nijdy~s. The 

NlLlMTB provides for the long-term cryogenic preservation of tissues usiing standard plrotoicols for 

collection and archival.The Stranding Network consist of regiolnal teams that respond to the 

stranding of marine mammals and are equipped to collect biological information and samples that 

can be used to understand the healtih, population dynamics, and life histories of marine mammals. 

The Monitoring Component conducts routine analysis of tissues in order to determine the lbaseline 

concentrations of chemical contanninants, biochemical componcmts, and biotoxins in marine 

mammals. The Quality Assurance Component is a program that insures a.ccuriicy, precision, level of 

detection, and intercomparability of data resulting from chemical analyses of marine mammals 

tissues. This program consists of interlaboratory comparison exercises and the plroduction and 

distribution of control and standard reference materials for chemicz~l analysis.. 

The MMHSRP includes marine manunal species from all marine waters of the United States. In the 

case of Alaska, the Program is being conducted through the Alaska Marine Ivlarnmal Tissue 

Archival Project, which is sponsored by the Minerals Management Service, U. S. Delpartment of the 

Interior. In order to provide a mechanism for consultation ancl coo~rdination ~ritl.~ recogmized 

scientists in the fields of research pertinent to the MMHSRP, the program has two scientific 

working groups who provide input during program planning: the National Marine Mammal Tissue 

B,ank Team of Scientists and the Task Group on Unusual Marine Mammal Mortalities. 

Because of the costs associated with collection and archival of specimens in the NMMTB and the 

increasing value of these specimens after years of storage, a formil Tissue Access Policy lhas been 

established to guide the release of specimens for research. A computer-based Informafiion and Data 

Management System is in the pla~nning stage. It will consist of several components including: 

sample, analytical data, and biological information tracking, as well as report bibliography and a 

specimen banks database. Additional avenues of research and development which are being 

clonsidered for the Program are a blcwd serum bank, the development of biotoxin methodologies, and 

tlne evaluation of biomarker technialues for marine mammals. 
Preface 

This Program Development Plan is the basic document setting forth a multi--ye,ar program to 

develop the organizaitional and physical resources and a database that can be used to facilitate a 

better understanding of the effects of human activity on the health of marine maxr~mals and their 

environment. Presented is the basis of planning and implen3entation strategy Ibr the Marine 

Mammal Health and Stranding Response Program conducted by the National Marine Fisheries 

Service (NMFS) Office) of Protected Resources. Acronyms used in this document are listed below: 

AMMTAP - Alasksl Marine Mammal Tissue Archival Project 

ECD - Enviro~nmental Conservation Division (of Northwest Fisheries Scienccsb Center) 

MMHSRP - Marine Mammal Health and Stranding Response I'rogram 

MMS - Minenals Management Service 

NBSB - National Biomonitoring Specimen Bank 

NIST - National Institute of Standards and Technology 

NMFS - Natilonal Marine Fisheries Service 

NMMTB - National Marine Mammal Tissue Bank 

NMMTB&SN - National Marine Mammal Tissue Bank and Stranding Network 

NOAA - National Oceanic and Atmospheric Administriition 

NWFSC - Northwest Fisheries Science Center 
Introduction 

Background 

Hundreds of marine marnmals are stranded each year on ithe bleaches of the United States. 

Apparently most of these are animals that die at sea and then are carried onshorc:~ by wilnds and 

currents. In the conliguou~s United Sates most of the stranded cetaceans are bolttlenose dolphins, 

Tursiops truncatus; pygmy sperm whales, Kogia breviceps; harbor porpoises, Phocoena phocotpna; 

and cornmon dolphins, Delphinus delphis. Stranded pimipetis consist mostly of' California sea 

lions, Zalophus californiarnus; harbor seals, Phoca vitulina; and northern elephaint seals, Mirorrnga 

angustirostris (Wilkinson, 1991). 

NOAA's National Marine Fisheries Service (NMFS) has established a marine nnarnmal st~anding 

network in each coastal region. The network consists of volunteers who respond to strandings of both 

dead antd live animals to gather basic biological information and, in the case of live stranclings, to 

attempt to rehabilitate and return the animals to the sea. At a minimum, the stranding network 

has beelm required to obtain info~mation on location of stranding, species, length, condition, and sex 

of the stranded animals. 'This data is contributing to the developmerit of a baseline1 of infc~rmation 

for detecting unusual mortality events. To coordinate stranding network activilries and ensure 

consistency among the Regions, a National Marine Mammal Stranding Network C:oordinator has 

been appointed with~ln the Office of Protected Resources. 

In 1987-88 a massive mortality of bottlenose dolphins occurried on the east coast of the Unuted 

States. Although the red tide organism, Gyrnnodiniurn breve, was implicated as a factor in causing 

this die-off, high levels olf organochlorine contaminants (PCB!j, chlorinated pe!jticides, etc.) were 

also found in these anim~ls (Geraci, 1989). It was not possiblc to determine whetlher or not these 

contaminants had any role in the die-off. Since no comparalble data on chemical contaminant 

concentrations in marine inlammals existed for the United States and there was no way to determine 

whether those levels foun~d in the bottlenose dolphins were "normal". It was alpparent that there 

was a real need for establishing the baseline levels of anthropogenic contaminants in marineh 

mamm,als in the Unitedl States and world-wide. It was also acknowledged that additional 

biological data should be collected by the stranding network., beyond that usually required andl 

that stranding network procedlures should be refined and standardized for all regions. This 

additional data should provide information which can be useful in monitoring the health of 

populaltions, including conitaminant burdens. 

The molnitoring of contaminant burdens in marine mammals, if carried out over many years, requires 

that one consider the possibility of the appearance of "new" presently unknown contaminants and 

the future developmend of new and better analytical techniques. Analytical ct- emi is try, 

particularly analytical organic chemistry, is a rapidly developing fl~eld. Methods are comtinwally . improving in accuracy, precision, and the ability to identify and separate closely related 

compounds. 

Data from samples analyzed today may be obsolete and have limited usefulness for future 

considerations. It is therefore necessary that during the design of a contaminant monitoring 

program or a baseline da~tabase, some system for carefully collecting and storing environmental1 

samples for future retrospective analysis be included. Environmental specimen banks establishetl 

for such purposes are in operation in Canada, Europe, and the United States, and many other 

countries are plamilng such systems. The German environmental specimen banlk s!ystem clonsists of 

two banks, one for human samples located at the University of Muenster and the second for otheLr 

types of environmental samples located at the Nuclear Research Center (KFA) in Jiilich 

(Boehringer, 1988). Botlh banks have operated for many years. The Canadian Wildlife Service 
maintains a large specimen bank in Hull, Quebec. This system has been in operation for many years 

and has been an integral part of contaminant monitoring in Canadian wildlife (Elliott et al., 1988). 

The present Swedish system is for wildlife research and a bank for human toxicological research is 

being plamed. Other banks areb located in Denmark, Finland, and Norway, and the International 

Atomic Energy Agency is evalu~ating what role it might play in establishing similar facilities for 

use by developing countries in e,astem Europe, the former Soviet Union states and countries in ,4sia 

and South America. 

Within the United States, the National Biomonitoring Specimen Bank (NBSB), which is llocated 

at the National Institute of Standards and Technology (NIST), is designed to archive and store a 

wide variety of environmental samples under cryogenic conditions for long periods of time 

(decades) The NBSB has supported environmental research and monitoring programs of several 

agencies including: the Food and Drug Administration (FDA), Environmental Protection ,Agency 

(EPA), National Cancer Institute (NCI), National Oceanic and Atmospheri~c Administra~tion 

(NOAA), and the Minerals Management Service (MMS) (Wise et al., 1988) 

Prograim Development 

In 1989, the NMFS Office of Protected Resources initiated the development of the National Marine 

Mammal Tissue Bank (NMMTB) at the NBSB, NIST. The NMMTB is designed to contain selevcted 

marine mammal tissues that have been collected and processed using rigorous well-documented 

protocols. These tissue samples are stored under the best conditions currently available (-150 "C:) for 

maintaining sampk? integrity. 

Enhanced support for the Marine Mammal Stranding Network also began in 1989. As part c11f its 

activities, the stranding network collects tissues for anthropogenic contaminant analyses andl the 

NMMTB. Additional sources of anirrials are those caught incidentally during comn~ercial fishing 

and animals taken for subsistence by Alaska native hunters. 

Early in tihe development of the program, NMFS established a Team of Scientists, with expertise 

in the fields of marine mammal biology, toxicology, chemical contaminants, and specimen banking, 

to serve an advisory role to the NMMTB. Through the efforts of the Teanil of Scientists a prol~ocol 

for collection, preparation and storage of tissue samples was developed based on an existing 

protocol that had been developed far a sirnilar program in Alaska, the kilaska~ Marine Mammal 

Tissue Archival Project (AMMTAP) (Ikcker et al., 1988,1991). In addition, 'the Team recornn~erided 

(1) the tissues most appropriate for specimen banking (liver and blubber), (2) the selection of sptxies 

to be sampled, (3) items to consider in a tissue access policy, (4) the development of a staindlard, 

"general" protocol which could be used by recognized research facilit,ies involved in mairine 

mammal research, (5) an inventory and location of tissues collected (samplle tracking systern), and 

(6) the development of a database that would include information on the tissues collected for the 

NMMTB. 

A demonstration phase (Pilot Program) was initiated in 1990 to evaluate the practical aspects of 

obtaining suitable samples for the NMMTB from both incidental catches and strandings. This 

operation was conducted on the northeast coast of the United States in cooperation with scientists 

at the New England Aquarium. The species sampled were harbor porpoiscb, Phocoenu phocoena (a 

coastal species), from incidental catches,, and pilot whale, Globicephnla mrelaena (a pelagic 

species), lfrom mass strandings. Based on the successful sampling of these two species in the 

northeast, it was decided that the operation in this region should continue with the addition of the 

Atlantic white-sided dolphin, L.agenorhynchus acutus, from mass strandings as a target sperk~s. It 

was also decided to expand the program to the southeast coast of the United States and the Pacific 

coast. The NMMTB presently contains 30 samples of blubber and liver tissues collected frorr~ six 
harbor porpoise and nine pilot whales. 

In 1991 NMFS combined the NMMTB and the Marine Mammal Stranding Neltwork into1 a single 

broader program anid added both a contaminant monitoring and a quality assurance component. 

This larger program was named the National Marine Mammal Tissue Bank. and Stranding 

Network Program (NMMTB & SN Program). The monitoring component, conducted by the 

Environmental Conwrvation Division (ECD) of the Northwest Fishmies Science Center (IWFSC), 

NMFS, includes real-time analyses to determine current levels (based on curnE1nt methods) of 

biotoxins and chemical contaminants in marine mammals, as well as research on ways to enhance 

methods and procedures used by the program. The quality assurance component, conducted by the 

Chemical Science antd Technology Laboratory, MST, includes the dlevelopment arrd application of 

quality assurance procedures and materials (such as SRMs) for chemical anz~lysis of marine 

mammal tissues. 

The NMMTB & SN Program has coordinated its work in Alaska with the AMMTAP, a project 

funded by the MMS, U.S. Department of the Interior. AMMTAP ha:; been archiving samples at the 

NBSB from marine lmammals taken in Alaska Native subsistence hunts. During these collections, 

the AMMTAP periodically has provided subsamples to the contaminant monitorin~g component of 

the Program. A closer coordination of AMMTAP with the Prograrn came about iin Janu,ary 1992, 

when the management of AMMTAP was transferred from the Office of Olcean Resources 

Conservation and Pissessment, National Ocean Service, NOAA, to the NMFS Office of ]Protected 

Resources. 

On January 3, 1992, U.S. Congress passed the Oceans Act of' 1992. This legislation amends the 

Marine Mammal Pr~otection Act by adding Title I11 which estatblishes the Marine ]Mammal Health 

and Stranding Response1 Program the purposes of which areb to: (1) facilitate the collection and 

dissemination of reference data on the health of marine mammals and health t~rends in marine 

mammal populations in the wild; (2) correlate the health of nnarine marnm~als and marine 

populations, in the wild,, with available data on physical, chemical, and biologici31l envinonmental 

parameters; and (3) coordlinate effective responses to unusual mortality events. Most of the items 

required in Title I11 are already in place in the NMMTB & SN Program. However, some additional 

requirements were stated, particularly regarding information /data1 management and contingency 

planning for major strantlings. The ability to respond to these additional requiremt:nts is uncertain, 

since funding for the program has not been appropriated. 

In response to Title 111, the NMMTB & SN Program has now been tlesignatecl as thle Marine 

Mammal Health and Stranding Response Program (MMHSRI'). This documen~t outlines the basic 

program within limitations of present funding. Appropriatilons will be required to expand this 

basic program to cover adlequately all requirements of Title 111. 

Authority -. 

The authority for NOA14, NMFS, to conduct the MMHSKP is provided in Marine Mammal 

Protection Act of 19172, 16 U.S.C. 1361-1407, as amended (Title 111) bly the Ocearls Act of 1992. The 

primary purpose of this legislation is to protect and preserve marine mammals in order to maintain 

the health and stability a~f the marine ecosystem as a whole. NOPiA's National Marine Fisheries 

Service has the responsi~bility for enforcing this act, except for manatees, polar bears, sea otters, 

and walrus, which are the responsibility of the U.S. Fish ancl Wildlife Service. 
Program Components 

The MMHSRP consists of four components: 1) Specimen Bank, 2) Stranding Network, 3) Monitoring, 

and 4) Quality Assu~rance. The goals of each of these components are discussed below. 

Specimen Bank 

Goal: Provide a resource that can be used 

for future retrospective analyses and 

documenlation of long-term trends in environmental quality 

The National Marine Mammal Tissue Bank (NMMTB) is maintained at the N,ational 

Biomonitoring Specimen Bank (NBSB), NIST, Gaithersburg, MD. The purpose of the NMMTl3 is 

the long-term preservation of specimens that are representative of the state of the animal 

immediat~ely prior to death. It is necessary that samples be collected, processed: and storedl under 

 condition,^ that avolid or minimize contamination or other changes in the chemical compositialn of 

the specimen. 

The sample collection and archival protocols for the NMMTB (see Appendix I[, pages 25-32) are 

consistent with the standard procedures of the NBSB which focus on collection^ of duplicate 

samples, use of m,aterials that minimize c~ontamination from any contact with the1 sample, and 

freezing samples in liquid nitrogen (LN2) as soon as possible after collection. A sample amount of 

about 150 g was selected as sufificient quantity of material to allow multiple anlalyses over a long 

period of time. The collection of two equivalent samples of 150 g provides one specimen for analiyses 

as required and onei for more permanent storage. Materials such as titanium1 and 'Teflon are used for 

all contact with the sample to minimize potential sample contamination. During sarnple 

preparation, contact with the sptximen is generally limited to clean, dust-free Teflon surfaces. The 

samples are stored at the NBSB facility, NIST, in LN2 vapor phase freezers, at -150 OC'. 'This 

temperature appears to be the best for long-term storage (Wise et al., 1989). 

Liver and blubber are the two principal tissues chosen for routine banking. Blublber, clue to its high 

lipid content, conct!ntrates several organic toxicants to relatively high levells. The liver is a ni~ajor 

site for detoxifying chemical contaminants and has a relatively high 111pid content, thus, this 

tissue also accumulates chemical contaminants and may retain relatively high levels of lheir 

metabolites. 

The archival of a representative collection of well preserved and documented marine m~annmal 

tissues will provide scientists with samples that have been collected and stored using standardiized 

procedures so that results for samples collected at various times can be compared to deternnine 

whether environmental trends exist. Scientists will also have the apporltunity to conduct 

retrospeciive analyses using improved analytical techniques or to measlure currently unknown 

contarniniints. 

The NMhITl3 is not intended to serve as a substitute for activities that are currcmtly being carried 

out by independent contaminants researchers. It is assumed that other tissues will continue flo be 

collected, banked, and analyzed for angoing monitoring and research. The NMMTB will not Itlave 

the capacity to displace such activities and should not be viewed as a primary source of samlple!; for 

real-time analyses. However, by providing protocols and quality assurance measures, the hlMlMTB 

can improve ongoing activities and provide valuable material for future retrospective study. 
Stranding Network 

Goal: Upgrade the capacity 

of the regional strancling networks 

Research conducted on stranded marine mammals has been the source of a significant part of our 

knowledge on marine mannrnals. Data collected from stranded animals can serve ar a baseiline from 

which comparisons can be made. Marine Mammal Stranding Networks have been set up within 

each of the NMFS Regions. The Networks are primarily composed of volunteers who hold LeHers 

of Authorization issued by the NMFS Regional Offices. They inclutde marine mammal scientists, 

aquarium persomeli, wildlife rehabilitation specialists, veterinarians, and wildlife management 

specialists. Network mernbers are required to collect basic biological information including the 

species, sex, length, and evidence of human interaction. Many of the Network members collect 

additional information and provide tissues to scientists for research. Informaltion collected from 

stranded animals can conlribute to an understanding of marine mammal life histories, population 

dynamics, and diseases nffectrng marine mammal populations, to name just few avenues of 

biological research. 

A program review of stranding network operations concluded that with a lilttleb effort, baseline 

data could be impro~red and that stranded animals could be the source of even more information. To 

realize the full potential of the Stranding Networks, an effort needs to be made to ensure that 

Network members know how to collect tissues for various ana1:yses and that the!{ have the required 

equipment. 

Stranded animals are currently the most accessible source of tissues from marine mamnnals, and 

numerous researchers utilize the tissues using a variety of protocols. In an effort to improve baseline 

data collected from stranded animals, NMFS has published lone technical document far use by 

Stranding Network rnernbers an~d is preparing a second. The first is a handbook for marine manimal 

strandings (Geraci and I,ounsbury, 1993). It contains a species identification key, treatment 

protocols for live stranded animals, and basic information on tissue collection. 

The second documeint is a forenisc laboratory manual for dolphins. This manual, which is being 

prepared by the NMFS 5;outhc:ast Fisheries Science Center, will address a variety of scientific 

disciplines inchding methodologies for life history, gross necropsy, histopathology, pathogen 

detection, and collection of tissues for contaminant and toxicological analysis. 

To enhance the capabilities of the Networks, the Program funcling will provide equipment, 

supplies, and protocol training. 

Goal: Detect and determine the cause 

of unusual marine mlamnxal mortalities 

To respond adequately to unusual mortality events, a systematic approach aovering a rang,e of 

possible causes needs to be developed. Available resources and gaps in either knowledge or 

Network coverage need to be determined in advance. As a first step in tlhis process, NMFS 

established a Task Group on Unusual Marine Mammal Mortalities in 1991. This gnoup is composed 

of experts from a numbt?r of scientific disciplines. They have esta~blished a series of criteria for 

determining when am unusual mortality is occurring based on information collected from marine 

mammal strandings. The 'Task Group was consulted five times between April 1991, and April 1992. 

Funds are required to respond adequately to such events. In the past,, it has been necessary to 

reprogram funds from base programs. In order to avoid repragramlming, it is tlie goal of NMFS to 

establish a permanent conltingency fund that can be used when needed. 
Goal: Maxiimize the use of 

the Regional Stranding Network 

data for management decisions 

As the collection of baseline data improves, information from strandings can be ulsed to understand 

the cause of mortalities, detect unlusual mortalities, and understand fisheries interaction. 

Additional research can help to determine stock definitions, population dyna~mics, and genetic 

information. In order to make use of silch data, the data collected must be more neliable, consiistent, 

and comparable between regions. Upgading the Regional Stranding Networks and combining the 

Network E'rogram with the National Marine Mammal Tissue Bank Prograrn should help improve 

this data collection. 

Monitoring 

Goal: Determine the baseline concentrations 

of chemical1 contaminants, bioche!mical 

componeniia and biotoxins in mafine maimmals 

High concentrations of persistent toxic substances in marine mammals is a phenomenon that is of'ten 

reported in the literature. These substances include both naturally occurring toxicants, sucl-L as 

heavy metals, as well as anthropogenic substances, such as polychlorinated biphenyls (PCBs) imd 

chlorinated hydrocarbon pesticides. Evidence to support the relatiionshilp between high 

contaminant levels in body tissues and detrimental effects to these animals has bet?n shown in a few 

cases (DeLong et al., 1973; Helle, 1980, 1981; Bergrnan et al., 1981; Reijnders, 1980, 1984, 1986). 'I'he 

tendency of marine mammals to bioac:cumulate contaminants can be explained by several ifaclrors 

including: relative position in the food web, tendency to accumulate large energy reserves in the 

form of body fat, relatively long life span, and relative ability to metabolize and excrete toxic 

substances (Tanabe et al., 1988; VValker, 1983). 

Information on the baseline levels of environmental contaminants and toxins in marine mammal 

tissues is necessary in order to determine environmental trends related to the health of' these 

animals. The results of the real-time analyses conducted by the Monitoring C:omponent of the 

Program will add to the needed database on levels of various contaminants and toxins in different 

species of marine mammals. R.eal-time analyses will also help to answer questions abalut the 

effects of chemical contaminants andl toxias on different species as well as on the dispositio~n of 

contaminants and toxins among tissues. 

Marine mtmrnal sampling by the stranding rretworks, observer program, and others associated vvith 

the Progriam will be coordinated to ccollect tissues for banking and monitoring from the same 

animals. Following this approach, analytical results will be available for all the samples within 

the NMMTB, either as determined on actual subsamples from the bank (e.g., bly NIST as part of 

their analyses) or on samples from tlhe sarne animal as part of the monitoring component. As 

questions arise concerning long-term trends, new pollutants, or the validity of previous results, 

tissues frolm the NMMTB, which were collected years earlier, can be withdrawn to address these 

issues. 

The NMFlS Northwest Fisheries Science Center (NWFSC), Environmental Conservation Division 

(ECD), conducts the Monitoring Component of the Program. In addition to liver and blubber 

routinely collected for the NMM'IB, the ECL) analyzes other tissue and fluid1 samlnles including, but 

not restricted to: bile, kidney, and brain tissue. Analytes include: I) organic chlemici~ls and lipids 

for blubber, liver, and brain; 2) rnetalls for liver, kidney, and brain tissue; 3) estimate of meta~bolites 

of polycyclic aromatic hydrocarbon:;; and 4) xenobiotic-DNA adducts for liver tissue. Certain 
compounds appear to be particularly important contaminants in marine mamnqals. These include 

methylmercury, coplanar PCBs, chlordane isomers, and polychlorinated camphenes (P(3Cs). As 

part of the Monitoring (Zomponent, the ECD will evaluate analytical methods used for routine 

monitoring to detemnine concentrations of these substances as well as others of importance. 

Goal: Develop guidedines for 

minimizing tissue sample variability 

Selective partitioning of environmental contaminants within the tissues of mirine marrrmals 

can affect the variability of the resulting analytical data and may be important in determining 

what sample locations are best for estimating body burdens. Such bias nnust be identified and 

steps taken to design sampling protocols to minimize and remove such bias and ,variability from 

the analytical results. Studies, will be conducted to estimate the distribution of cheirnical 

constituents in marine mammal tissues and its effect on sampling ~(ariability and error, and to 

recommend tissue sampling procedures that provide the best samples for accuratc2ly determining 

the concentration of environmental contaminants in these tissues and providing the best 

estimate of body burdens. 

Before stranded animals can be fully utilized as a resource for contaminant analysis, a basic 

question that needs to be ,answered is how long an animal may be dlead before it is considered 

unsuitable for sampling for contaminants and other chemical ~on~stituents. Wlhat che~micall 

changes occur in tissues auld organs between the time of death and tissue collect lion^, and d~uring 

various conditions of storage prior to analysis? Such changes probably depend (on the chemical 

constituent in question, type of tissue, the physiological state of the aniimal at time of death, 

and environmental conditions. Studies will be conducted to determi,ne the tempored changes in1 

selected chemical constih~ents of marine mammal tissues following death and to dletermir~e the 

maximum time that can elapse between animal death, sample removal, and saniple freezing: 

before change in selected chemical constituents become significimt and do not reflect the oniginal 

conditions in the animal at time of death. 

Quality Assurance 

Goal: Insure the accuracy, precision, level 

of detection, and intercom~parability of data 

resulting from chemical ar~alyses 

of marine mammal tilssue samples 

As part of the MMI-ISW, analytical results for chemical contaminants in marine mammal tissues 

will be generated by labo~ratories in both the Monitoring and the Specimen Bank C:omponents. To 

provide accurate alnalytical data, a quality assurance (QA) program has b'een developed for 

assessing the accura~cy and comparability of results among the llaboratorie+s for b'oth the organic and' 

inorganic analyses. This QA program, which is administered by the Chemi~cal Science and 

Technology Laboratiory, IVIST, includes the following: 

1) Interlaboratory comparison exercises among 1aboratoric.s involved in NOAA praljects 

related to marine mammal tissue analyses. 

2) Preparation, analysis, and distribution of marine nlammal tissue control materials. 

3) Development (of Standard Reference Materials (SRM's) for use in the ar~alysis of marine 

mammal1 tissues. 

Following a comparison and evaluation of analytical methods for organic contaminants 

(particularly for lipid-rich tissue matrices) between NIST and the ECD, an interlaboratory 
comparison exercise was conducted. Participants included NIST, the ECD, and several other 

laboratories involved in measurements of chemical contaminants in marine marrunal tissue!:. 'I'his 

exercise consisted of the analyses of two "control" materials (whale blubber fc~r organic analysis 

and whal~e liver for inorganic analysis) prepared by NIST from tissues obtained from two pilot 

whales. Approximately 2 kg of each tissue type were prepared as frozen tissucc! hornogenates for 

distribution as subsamples of 15-20 g, each. The list of analytes for the first interlaboratory 

comparison exercise was based on input from researchers involved in chemical contaimir~ant 

measurerr~ents in marine mammal tissues. 

Recommended reference values will be determined through continued analyses of these maiterials, 

which will then be distributed for use as control material during routine analyses of ma~rine 

mammal l5ssues. The experience gained from the analyses of the control materialls is being used by 

NIST to clevelop both blubber end liver Standard Reference Materials (SRMs) for use in marine 

mammal tissue analysis. An SEW is a homogeneous material that has been analyzed by two or 

more indc2pendent analytical methocis to certify concentrations of selected organic' or inorganic 

constituei~ts. These SRM's will then1 be available to any researchers involved in the analysis of 

marine mammal tissue for use ir~ the validation of analytical methods. 
Target Species 

The NMMTB protocol is designed to collect a few selected tissues from a limited number of 

representative, "inclicator" species. Since each region of tlhe United States is unique in the 

potential availability of acceptable marine mammal samples, the specific species to be sampled 

for the Tissue Bank are determined on a regional basis. The selection of the "indicator"species has 

been based on the following considerations: 

1) Available source of a large number of animals on a regular (ideally amrrual) basis; 

2) Representatives of both coastal and pelagic species are included; 

3) Representatives of both cetaceans and pinnipeds are included; 

4) Potentiall for accumulating anthropogenic contamirlants to relatively high levels; 

5) Trophic levels should be considered (bottom feeder, pelagic fish feeder, pelagic 

plankton feeder); and 

6) Relative value as fad or subsistence species for humans (applicable to Alaska). 

Indicator species selected for the NMMTB based on these consiclerations are summarized irr Table 1. 

This species list may expand in the future, since Title I11 of the Marine Mammzrl Protection Act 

requires coordir~ation with the WSFWS. Four marine mammal spec~~es are under the management 

authority of the USFWS manatee, Trichechus manatus; walrus, Odobenus rosrnarus; sea otter, 

Enhydra lutris; and polar bear, Ursus maritirnus. The manatee could functron as an i~ndicator species 

in the southeast United States, walrus and polar bear in Allaska, and sea otter in Alaska and 

Pacific Coast. Manatee, walrus, and polar bear are of particular interest since they represent 

trophic levels not represented in the species given in Table 1. 
Table 1.-Indicator species for the National Marine Mammal Tissue Bank.1 

Region Coastal Pelagic 

Northeast 

Southeast 

Pacific 

Alaska 

Harbor porpoise, 

Pfrocoena phocoena (I) 

Pilot whale, 

Globicephala malaena i(S) 

H<arbor seal, 

Pilloca aitul ina (I) 

*Atlantic white-sided dolphin, 

Lagenorhynchus acutus (S) 

Bo ttlennse dolphin, 

Tursiops: truncatus (S) 

Harbor porpoise, 

Pliocoena phocoena (I) 

Harbor seal, 

Plzoca vitulina (I, N) 

Califorr~ia sea lion, 

Zalophus californianus (I) 

Ringed seal, 

Plzoca hispida (N) 

Pilot whale, 

Globicephala malaena r(S) 

Dall porpoise, 

Phocoe~oides dalli (I) 

Northern fur seal, 

Callorhinus zrrsinus (N) 

Northelm fur seal, 

Callorhinus ursinllts (N) 

Beluga whale, Bowhead whale, 

Dmelphin,apterus leucas (N) Balaenra mys; ticetus (N) 

lNorthea,st = northeastern U.S., Souttheast = southeastern U.S. (Atlantic andl Gulf of Me:~:ico), 

Pacific (including the Gulf of AJaska), Alaska = Bering Sea and Arctic Ocean; I[ = incidental catch, 

S = stran'ding, N = Native subiiistencce harvest. * = considered to be both a ca~astal, and a pelagic 

species. 
Management Plan 

Organization 

The diverse expertise and resources required by the MMHSRP has made it necessary that several 

NMFS organizationls as well as organizations outside of NMFS be involved in the conduct of the 

Program. The primiary organizations involved in the Program are: the NMFS Office of Protected 

Resources, the NMFS Environmental Conservation Division (EClD) of the Northwest Fisheries 

Science Center (W'FSC), the NMFS regional offices and their associated strandimg nehvorks and 

observer programs, and Chemical Science and Technology Lalboratory of the National Institute of 

Standards and Technolo&y (NIST). The responsibilities of each of these prima~y organizations are 

reflected in the Program structure which is organized to atldress six basic functions: program 

management, response to mortality events, necropsies and sampling, contamini+nl and biochemical 

monitoring, quality assurance, and specimen banking (Table 2). 

Program management is the responsibility of the NMFS Office of l'rotected l7:esc)urces. Program 

management includles establishing program policy, overall planning, budget formulations and 

allocations, coordinatioin of program activities including international coordination and 

cooperation, establisihing and maintaining a system for informtation and data management, and the 

dissemination of prclgrarn information and results to the scientific co~rnmunity, govcrrnment: agencies, 

educational organiziations, and the general public. The Office of Protected Resourres will maintain 

a Program Manager and associated staff for facilitating these activi~ties. 

Response to morta~lity elvents is the responsibility of the NMFS regional offices through their 

regional stranding networks. The regional stranding networks include particlipa~nts from various 

universities and non-governmental organizations. The interface between the Program and the 

regional stranding networks is provided through the Regional I3tranding Network Coordinators and 

the National Stranding Network Coordinator located in the program^ Office. On~e alf the mLajor goals 

of the Program is to upgra~de the capacity of the regional strandling networks to respond tai mortality 

events. Therefore, a very important responsibility of Program management will be to provide 

resources and trainir~g necessary for the stranding networks to srlccessfully perform their function. 

Necropsies and sampling for the NMMTB and the monitoring component of the Progam is ]performed 

through arrangements made with the NMFS Observer Prc~gram~ and several indivicluals and 

organizations opera~ting under the authority of the regional strantding network.^. The interface 

between the Progra~m Manager and the regional stranding networks and observer programs is 

provided by the Regional Stranding Network Coordinators andl Observer Progre~m Coordi~nators. In 

the case of Alaska,, other agencies, local researchers, and management organ~izations provide 

additional necropsy and sampling support for animals taken during Alaska native subsistence 

harvests. These organizations presently include the MMS Alaska Office, ithe NMFS Western 

Alaska Field Office, the North Slope Borough Department of Wildlife Management, the Eskimo' 

Walrus Commission, and Kawerak, Inc. 

Contaminant and biochemical monitoring is conducted by the NMFS, NWFSC, ECD. 'This work 

requires careful coc~rdination and cooperation with the specimen banking and quality assurance 

activities of NIST. Collitboration with similar monitoring programs being condlucted lby marine 

mammal investigators from other countries is encouraged in order to insure comparability of 

analytical data and to develop comprehensive research approaches. 

Quality assurance for chemical analyses is provided by NIS'T, Chemical Sciemct! and Technlology 

Laboratory, through interlaboratory comparison exercises with the ECD and other investigators 
involved in research on contami~nants in marine mammals, as well as the production and distriblution 

of standard reference materials :specifically designed for marine mammal tissue analyses. 

Specimen banking for the Program is the responsibility of the National Biomonitoring Slpecimen 

Bank (NEISB) at NIST. The NEISB uses standard and state-of-the-art prcceduires for maintaining 

environmental samples under cryogenic conditions for long periods of time and routinely monitol-s the 

conditions of the samples during; storage. 

Table 2.--Organization structure of the National Marine Mammal Tissue Bank and Stranding 

Network Program, 

Functions Performing organization 

Program ]Management 

Response to Mortality Events 

Necropsies and Sampling 

Contamirrant and Biochemical 

Monitoring 

Quality lissurance 

Specimen Banking 

National Marine Fisheries Service 

Office of Protected Resources' 

Silver Spring, Maryland 

National Marine Fisheries Service 

Regional Offices throu:gh their 

Regional Stranding Neltw0rk.s 

& 

National Marine Fishelries Service 

Office of Protected Resources; 

National Stranding Network 

Coordinator, Silver Spring, jMaryland 

National Marine Fisheries Service 

Regional Offices through their 

Regional Stranding Neitworks and 

Fishery Observer Programs; 

Other organizations 

National Marine Fisheries !Service 

Northwest Fisheries *ience Center 

Environmental Consenration, 

Division, Seattle, Washington 

National Institute of Standards and 

Technology, Chemical Science and 

Technology Laboratory, Gaiithersburg 

Maryland 

National Institute of S~tandards and 

Technology, National Biomonitoring 

Specimen Bank, Gaithersburg, 

Maryland 
Program Planning and Review 

An important part of the Program management is consultation and coordination of these efforts with 

recognized scientists in the fields of research pertinent to the MMHSRI'. PL mechanism for 

achieving this ,coordination is the formation of scientific worlting groups to provide input during 

program planning. The IJrogram has two such groups: the National Marine bIanunal Tissue Bank 

Team of Scientists and the Task Group on Unusual Marine Marnmal Mortalities. 

The National Marine Mammal Tissue Bank Team of Scientists was formed early irii the development 

of the Program (19119) to insure peer review of all Program activities. The Team, m~~eets on an annual 

basis to review program activities of the previous year and to discuss proposed wolrk for the coming 

year. The Team consists of scientists in marine mammal biological research and rnamagemelnt, marine 

mammal physiology, to~xicology, chemical contaminant research, ecology, anallytical chemistry, 

and environmental ,specinlen banking (Table 3). 

The Task Group on Unnsual Marine Mammal Mortalities, which was set up ]in 11991, is consulted 

when an unusual nnortallity event is suspected. An effort was madle to includ~e in the Task Group 

scientists from a nu~mber of different specialties including: epidemiology., veterinary pathology, life 

history, toxicology, anti detection and treatment of diseases in marine marnmals (Table 4). 

Specialists for both pinnipeds and cetaceans and individuals familiar with strandings are included. 

The Task Group has established a series of criteria for determining whether an un~usual mortality is 

occurring. They provide suggestions and guidance during the course of investigations and review the 

results of studies. 

Tissue Access Policy 

Because of the costs associated with the collection and archival of specimens in the NMMTB and 

the increasing value of these specimens after years of storage, an access policy has been established 

that addresses the disposition of the banked tissues and the procedures, justific;rtion, a~nd review 

process required for access to this resource. The "NMMTB Acciess Policy" is provided in Appendix I, 

pages 33-35. 

Information and 

Data Management 

A computer-based lhforrrration and Data Management System will be established for the 1MMHSRP. 

Before the passageb of Title 111, the following were identified as requiring suclh a system to be 

established anti maintained by the NMFS Office of Protectecl Resources in support of the Tissue 

Bank: sample tracking, analytical data tracking, supplemental information tracking, repor4 

bibliography, and specinnen banks database. 

Sample tracking would be facilitated through a continuously updated database which in'dicates the 

location and fate of each sample collected by the Program folr ard~ival. This database would also 

track those samples collected from the same animals for other research purposes. 

Analytical data tracking, would include the identification and descriptions a~f e~nalyses that have 

been conducted on samlples collected by the Program and would provide information that would 

enable one to access this data. (e.g., identified reports, publications and journal akrticles, or 

individuals and organizations having this data, if unpublished). 
Table 3. -Members of the National Marine Mammal Tissue Bank Team of Scientists. 

Member Affiliation 

Howard 7N. Braham 

John Calambokidis 

Greg Early 

Sylvia G,alloway 

Joseph R. Geraci 

Romona I-laebler 

Robert J. Ilofman 

Linda Lowenstine 

James Mead 

Daniel K. Ode11 

Usha Varanasi, Chair 

Stephen ,4. Wise 

National Marine Mamrnal Liaboratory 

National Marine Fisheries Service 

Seattle, WA 

Cascadia Research Collective 

Olympia, WA 

New England Aquariurn Central Wharf 

Boston, MA 

Southeast Region 

National Marine Fisheries Sbervice 

Charleston, SC 

Ontario Veterinary Colllege 

University of Guelph 

Guelph, Ontario, Canada 

USEPA 

Narrangansett, RI 

Marine Mammal Commissior~ 

Washington, DC 

School of Veterinary M:edicime 

University of California - Davis 

Davis, CA 

National Museum of Natural History 

Smithsonian Institutiori 

Washington, DC 

Sea World of Florida 

Orlando, FL 

Environmental Consenration Division 

Northwest Fisheries Science Center 

National Marine Fisheries !iervice 

Seattle, WA 

Chemical Science & Te:hnolc)gy lab. 

National Institute of Standards & Technology 

Gaithersburg, MD 
Table 4. -Members of tl~e Task Gmup on Unusual Marine Mammal Mortalities. 

.- ------- 

Member Affiliation 

---- .- 

Gregory D. Bossart Miami Seaquarium, Miami, F1, 

Leslie Dierauf Washington, DC 

Joseph R. Geraci, Chair Ontario Veterinary College, University of Guelph 

Guelph, Ontario, Canada 

Romona Haebler USEPA, Narrangansett, RI 

Robert J. Hofrnan Marine Mammal Commission, W,ashington, LC 

Jeff Horwathl USFWS, Arlington(, VA 

Thomas Lipscomb 

James Mead 

Thomas O'Sh~ea 

John Reif 

Thornas Roffe 

Armed Forces Institute of Path~ology, 

Walter Reed Arm37 Medical Center 

Washington, DC 

National Mu:jeum of Natural History 

Smithsonian Institution, Wasl~in;gton, D1.C. 

National Ecalogy Kesearch Genber, USFWS 

Fort Collins, CO 

Department of Environmental Health 

Colorado State Un~iversity, Fort Collins, CO 

National Wildlife Health Research, Center 

USFWS, Madison, WI 

Linda Schlater National Veterinary Services Labs, Ames, Vi 

Randall Wells Mote Marine Laboratory, Sarasota, FL 

Dean Wilkinson,l Exealtive Secretary NMFS, Office of Pirotected Res;ou~rces 

Silver Spring; MD 

Thomas Williams Carmel, CA 

Kate Wynne University of Alaska Sea Grant Program 

Marine Advisory ]Program, Fa~irbanks, AK 

Nina M. Young Center for Marine Conservation, washing ton^, DC 

--- ___---- 

1 Nonmembe~r agency contad, 
Supplemental information tracking ~rould be facilitated through a database that identified all1 

supplemental information collected on the individual animals during sampling, including necr~opsy 

reports and other information obtained by stranding networks or researchers. 

Report bibliography would consist of abstract:; of all reports generated by th~e MMHSRP and oth~er 

research conclucted on the animals stampled by the Program. 

Specimen banks database would contain information on all applicable holdingls of tissue banks other 

than the NMIMTB. This informiltion would be useful to investigators interested in acquiring acccas to 

such samples, many of which do not require the stringent archival procedures used by the NMMTB. 

With the passage of Title 111, the i,nforntation and database requirements have been expanded 1.0 

include reference data on the health of nurine mammals and populations of marine mammals, arrd 

data on species of marine m~amrnals that are subject to unusual mortali~ty events. Witlhout 

additional f~tnding for the Progra~m, the Information and Data Management Sysltem can not I:,e 

expanded at this time. 

Internatior~al Coordination 

and Cooperation 

Although the1 MMHSRP is concerned witlh marine mammals of U.S. coastal waters, it is recognized 

that most, if not all, of the species of comcern are distributed throughout the world's oceans aritd 

many of the ]populations that occur in the' U.S. coastal waters migrate through the waters of several 

nations. Incidents of unusual Inass standings, die-offs, and occasions of pollu~tant incidents arc! n'ot 

occurring only within one national air even regional boundary, but is a world-wide phenomenon. It is, 

therefore, important that the Program actively pursue intemational coordina~tion ,3nd cooperation 

in the various avenues of monitoring and research conducted by the Program. 

The Program is presently andl will1 continue to be involved in the International Biomonitorirrg 

Specimen Banking Symposia and the International Committee on Environmental Specimen Banlung. 

This commitlee is involved in planning, organizing, and sponsoring the bian~nual symposia and is 

attempting tcl standardize technical requirements for environmental specimen banks. 

The Program, is also pursuing avenues of cooperative studies and information exchange with tt11e 

International Arctic Community through mechanisms established by the Arctic Research Policy Act 

(ARPA) of 1984 and the International Arctic Science Committee, as well as cooperative exchanges of 

samples and data set up by the AIVIMTAP with the Department of Fisheries and Oceans Canada. 

The Arctic is the recipient of nmny kinds of airborne contaminants from the lower latitudes. Die 

community of Arctic nations, which includes the United States, has many shared natural resources, 

specific populations of marine mammals, and common sociological and eco~r~omic consideratiorrs, 

including a iunique subsistence lifestyle that relies on certain species of marine mammals as a 

supplemental food source. It is therefore mandatory that, if the MMHSRI' includes the Arctic, 

collaboration and cooperation with the work of other Arctic nations be an integral part of' the 

Program. 

The Program will be pursuing means of cooperation and coordination with researchers in the 

Antarctic through the National Science Foundation's Polar Programs. Another avenue of 

intemational cooperation is through the Program's quality assurance (QA) ac~tivities. Investigators 

from Canada and Germany are already involved in this Program component. The exparrsion of the 

QA component will continue with the intent that eventually most of {.he marine marnm(al 

researchers of the world invo'lved in cl-remical contaminant analyses will be participants in th~e 

MMHSRP QA activities. 
The discussion presented above indicates only a few avenues for building intennational connections. 

Other avenues will undoubtedly appear as the Program progresses. 

Program Relports 

In addition to special technical reports that will be producedl by t:he program, (e.g., Handbook on 

Marine Mammal S!trandli~gs), the following reports will be pr~oduced by the Prog;:ram on an annual 

basis: 1) Annual Program Report, 2) Tissue Bank Report,, 3) Stranding Welbvork 'Report, 4) 

Contaminant Moniitoring Report, and 5) Quality Assurance Relport. 

The Annual Progriam Report will be produced by NMFS, Office of ]Protected Resources. 'Ihis report 

will contain a surnrnary of the results of other program components, descriptions of plamt?d projects, 

a description of any interagency and international cooperation or collabora~tion, a discussion of 

outreach activities, an updated bibliography of reports and publications generated by the program, 

an updated database report, and a presentation on budget expenditures. 

The Tissue Bank Report will be produced by NIST. It will plrovidle a description of all significant 

activities of the Tissue &ink and will include an inventory of archived specimens and the results of 

any chemical amalysis hit were performed for the monitoring of sa~mple integrity. 

The Stranding, Network Report will provide a general ovexview of the activities of each of the 

Regional Stranding Networks as related to enhanced network capabiliities,, other Program 

components (e.g., nnonitoring and tissue banking), and response to any unusual n~ortality events. This 

report will be produced by the National Stranding Network Coordinator with input from the 

Regional Straniding Network C:oordinators. 

The Contaminant Mordtoring Report will be produced by NMFS, WFSC, ECD. This will be a 

technical report providing analytical results of the Contaminant Monitoring Component along with 

appropriate interpretation of the data. 

The Quality Assuranc~e Report will be produced by NIST, with input from ECD. It will be a 

technical report providing the results of interlaboratory comparisons, SRM characterizations, etc. 

Outreach 

It is the policy of the MMHSRP to encourage and support the publication of data1 resulting from the 

program in scienb~fic peer-reviewed publications. However, with the interest of the general public 

in the environment and partvcularly in marine mammals, xt is apparent that a, variety of meap 

other than jolurn,al publications and annual program reports are required for disseminating 

information resulting from the Program to a broader audience. The interest4 parties will probably 

include the general publlic, industry, educational organizatians, and various pubdic interest t~oups, 

in addition to governmc2nt agencies and scientists. This will requircl the develolprnent of an "outreach 

strategy" for the program which will probably result in the production and distribution of several 

types of info~rmalion media, including newsletters, educat~~onal videotapes, general ~lnforrnation 

pamphlets, special technical reports, and popular articles for magazines and lay journal!:. The 

details of this "outreach strategy" is yet to be developed. 
]Program Milestones 

Presented below .are major milestones in the development of the MMHSRP . Additional details on 

specific projects being conducted by the pr~~gram components will be provided in the Annual Program 

Report . 

Specimen Bank 

Establish the NMMTB ...................................................................................................... 5/89 

Establish the iNMMTB Team of Ijcienlists ........................................................................ 12/89 

Initiate Pilot Program in NE ............................................................................................. 91'90 

Initiate NMM'TB Program in NE ...................................................................................... 10/91 

Initiate Pilot F'rogram on West C:oast ................................................................................. 5\92 

Initiate NMM'I'B Program on West Co. ast ........................................................................... 5/93 

Initiate Pilot Program in SE ................................................................................................. Initiate NMM'TB Program in SE .......................................................................................... 

Stranding Network 

Initiate Stranding Network enhancement procedures ....................................................... 10/'90 

Establish the Task Group on Unusual Marine Mammal Mortalities .................................... 3/91 

Monitoring 

Initiate the Cctntaminant Monitoring in the NE ................................................................ 3/91 

Initiate the Cctntarninant Monitoring in the SE ................................................................. 3/91 

Initiate the contaminant Monitoiring an the West Coast .................................................. 3/92 

Quality Assu~anc~e 

Initiate interla~boratory comparison exerc:ises ................................................................ 3/91 

Initiate Blubber SRM Development ..................................................................................... 10/91 

Initiate Liver SRM Developmenit ....................................................................................... 10/'92 

Management 

Initiate the development of a database nnanagement system .............................................. 5/'92 

Initiate the development of an "outreadh strategy" ............................................................ 5/'92 
Additional Research Requirements 

Not all of the analytical tools, research approaches, or basic biological and physiological 

information are reladily available for evaluating the significance of anthropogenic contaminant 

loads in marine mammals and for understanding the effects of human activity on the health of 

marine mammals and their environment. Many questions can only be addressed iJhrough long-term 

experimental research. For example, analysis of wildlife tissues or other sen~tirrel organisms can 

provide a long-term picture of the state of contamination integrated over area and time, its increase 

or decrease, and the arrival of new anthropogenic contaminants. However, the determunation of 

biological significaince of the contaminant concentrations relative to the health of the i~ndividual 

animals and the population requires the kind of data used lo construct ioxicokinetic. and 

ecotox~cological nlodels (Clark et al., 1987). Rates of uptaktb, metabolism, and rlearance must be 

understood so that the status of the organism with respect lo equilibrium or steady state can be 

determined. Feeding habits, migration or mobility, and changes in physiobgi~ral compartments 

(e.g., lipid) can influence concentrations in tissues; therefore, natural history, physiology, and 

ecology must also be understood. 

Much of this basic research is outside the scope of the MMHSIRP; however, two avenues of research 

and development which have been identified as being within the scope of the Program and are 

being considered for future addition are discussed below. 

Creation of a marine mammal blood serum bank. Within the llast fifteen years a number of 

epizootics have occurredl that have been caused by disease agents. In 1979-80, an unp~recedlented 

number of harbor seals stranded on Cape Cod. An influenza A virus was determined to be 

responsible (Geraci et all., 1982). In 1984, an epizootic caused by leptospirosis ocr:urred in the west 

coast California sea lion population (Dierauf et al., 1985). In 11988, lover 17,000 pilrlnipeds in Europe 

died as a result of phocine distemper virus (Osterhaus et al., 1990). Most recently a distemper virus 

has been implicated in the deaths of large numbers of striped dolphins in the 1~4editeriranea~n. In 

order to determine appropriate actions, it is important to know what diseases are endemic in a 

population and whether a disease is being introduced to an in~nun~ologically naive population. As 

an additional element in assessing the status of marine rnaw~mal health, it is, th~erefore,, necessary 

to monitor diseases affecting populations. 

A record of diseases may help in isolating specific pathogens. Dise,ase antibodiies can be detected in 

blood serum. In much the same way as retrospective anal,yses can be valuablle in cc~ntanunant 

analysis, examination of historical blood serum can providle information orn whether a disease 

agent has been recczntly introduced or is endemic in a population. As resources become availablle, the 

MMHSRP will set up blood serum banks that can be used for current analysis of diseases in marine 

mammal populati~ons antd for retrospective analysis. Both live animals and freshly dead animals 

can be used for the collection of blood serum samples. Blood serum can be frozen fior later analysis. 

Development of biotox:in methodologies. There is evidence that biotoxins cre!abid by algal blooms 

can affect marine mamrnals (Gilmartin, 1987; Geraci, 1989; Geraci et al., 1989; and O'Shea et al., 

1991). Stomach cointents from dead animals and analysis of pr'ey halve been the most common1:y used 

methods for detecling biiotoxins. As a long-term goal, the Program should include developing more 

direct method:$ for determining the presence and impact of biotoxins on rnarine mammals. 

Application of i~iomarker techniques to marine mammal studies. Determining the tissue 

concentrations and body burdens of selected contaminants in marine mammal tissues does not lead 

directly to a full underljtanding of the toxicological significance of the chernic,als detected. The 

developing field of biomarker measurements in wildlife toxicology shows future ~prorr~ise in

identifying physiologica~l responses that will provide inform~ation on exposulre lo a range of toxic 
compounds ;and to integrate toxicologi,c interactions from mixtures of chemical1 contaminamt!j 

(Peakall, 199:!). Currently the appllication of biomarkers to marine mammals is in the very early 

stages of development, and no biomarkers of contaminant exposure nor physiological response have 

been adequately validated for use with marine mammals. Further, the utility of biomarkers in 

evaluating mass mortality events is only beginning to be assessed. Thus the Plrogram shoulcll 

encourage further development and evaluation of biomarkers to complennent the techniques 

currently used in monitoring chemical contaminant exposure and response in marine rrmrnals. 
Acknowledgments 

Numerous individuals have been directly involved in the developmer~t of the Marine Mammal 

Health and Strandiing Response Program and have provided review comments on ithis report. 'They 

include the National Marine Mammal Tissue Bank Team of Scientists; Usha Varanasi and her staff 

at the National Marine Fisheries Service (NMFS), Northwelst Fisheries Science Center, 

Environmental Co~nserviation Division; Steve Wise and Barbara Koster, N,ational Institute of 

Standards and Teclmology; Gloria Thompson, NMFS Headquarters,; Bob Hofman, Marine Mammal 

Commission; and CY. L. Hobart, NMFS Scientific Publications Office. A special aclknowledgmc~nt is 

given to Nancy Fo!jter, WMFS Deputy Assistant Administrator, who provided the original concept 

that has developed into the program described in this documemt. 
Li.terature Cited 

Becker, P. R., S. A. Wise, B. J. Koster, and R. Zeisler. 1988. Alaskan Marine Mammal Tissu~e 

Archival Project: A project tiescription including collection protocols. U.S. Dep. Comer., 

Nat. Bur. Standards, Gaithersburg, Md, NBSIR 88-3750,46 p. 

Becker, P. IR., S. A. Wise, B. J. E:oster, and R. Zeisler. 1991. Alaska Marine bdammal Tissue 

Archival Project: Revised collection protocol. U.S. Dep. Commer., Nat. Iinst. Standarcls 

Technol., Gaithersburg, Md, NISTIIR 4529, 39 p. 

Bergman, A.,, M. Olsson, and L. Reutergardh. 1981. Lowered reproduction rate in seal popula.tic~n 

and PCB concentration. Int. CIounc. Explor. Sea, Copenhagen, C-m. 1981/N: 10. 

Boehringer, U. 1988. Ongoing activities of the Environmental Specimen Bar~tking Program in th~e 

Federal Republic of Germany. h S. A. Wise, R. Zeisler, and G. M. Goldstein (Editors,), 

Progress in environmental specimen b,anking. U.S. Dep. Commer., I\Jat. 13ur. Standard:;, 

Gaithersburg, Md, NBS Spec. Publ . 740, p. 1-9. 

Clark, T. P., F:. J. Norstrom, G. A. Fox, and H. T. Won. 1987. Dynamics of organochlorine compou~ndls 

in Herring Gulls (Lams argentatus): 11. A two-compartment model and data for Iten compound:;. 

Environ. Sci. Technol. 6547-559. 

DeLong, R. L,. , W. G. Gilmartin, anid J. C;.. Simpson. 1973. Premature births in California Sea-Lilonsi: 

Association with high organochlorine pollutants residue levels. Science 181:1168-1170. 

Dierauf, L. A., D. J. Vandenbroek, J.. Roletito, M. Koski, L. Arnaya, and L. J. Gage. 19815. An epiza~oti~c 

of leptospirosis in California :sea lioins. J. Am. Vet. Med. Assoc. 187(11):1145-1248. 

Elliott, J. E., R. J. Norstrom, S. W. Kennedy, arid G. A. Fox. 1988. Trends and effects of envirorrmental contaminants determi~ned from arialysis of archived wildlife samples. In S. A. Wise, 

R. Zeisler, and G. M. Goldstein (Edlitors), Progress in environmental sprximein banking. 'U.!:;. 

Dep. Cornmer., Nat. Bur. Startdard!~, Gaithersburg, Md, NBS Spec. Publ. 740, p. 131-142. 

Geraci, J. R. 1989. Clinical investigations of the 1987-88 mass mortality of bottl~enose dolphin~s 

along Ihe U.S. Central and South Atlantic coasts. Final rep. to National Miarine Fisheries 

Service; U.S. Navy Office of Naval Research. Univ. Guelph, Ontario Vet. Coll., Onta~rioi, 

63 P. 

Geraci, J. R., imd V. J. Lounsbury. 1993. Marine mammals ashore: A field guide for stranclings. 'Tex.. 

A&M 1Jniv. Sea Grant Publ. 93-601., Coll. Sta., 305 p. 

Geraci, J. R., D. J. St. Aubin, I. K. Barker, R. G. Webster, V. S. Hinshaw, W. J. Bean, H. L. Ruhnke, 1. 

H. Prescott, G. Early, A. S. Barker, S. Madoff, and T. R. Schooley. 198;!. Mass mortality of 

harbor seals: Pneumonia associatecl with influenza A virus. Science 215:1129-'1131. 

Gilmartin, W. G. 1987. Hawaiian monk: seal die-off response plan, a workshop report. U.S. Dep. 

Cornmer., NOAA, Nat. Mar. Fish. Srv., Cmuthwest Fish. Cent. Admin. Rep. HI-87-19, 55 lp. 

Helle, E. 1980. Lowered reprocluctive capacity in female ringed seals (Phoca hispida) in the 

Bothniiin Bay, northern Baltic Sea,, with special reference to uterine occ'lusio~ns. Ann. Ztwl. 

Fennici 17:147-158. 
Helle, E. 1981. Reproductive trends and occurrence of organoch10rine:j and heavy-metals in the 

Baltic seal populations. Int. Counc. Explor. Sea, Copenhagen, (1I.M. 1981 /lEP:!17, 

13 p. (mimeogr.). 

OfShea, T. J., G. B. :Rathblun, R. K. Bonde, C. D. Buergelt, and D. K. Odell. 1991. An epizootic of 

Florida manatees associated with a dinoflagellate bloom. Mar. Mamm. Ski. 7(2):16!5-179. 

Osterhaus, A. D. M. E., J. Groen, H. E. M. Spijkers, H. W. J. Broeders, HI. W. J. Uytde Haag, P. de 

Vries, J. S. Teppema, I. K. G. Visser, M. W. G. van de Elildt, and E. J. Vedtller. 1900. Mass 

mortality in seals caused by a newly discovered virus-like morbillivir~ls. Vet. Microbial. 

23343-350. 

Peakall, D. 1992. Animal biomarkers as poll~aaion indicators. Ecotoxicol. Ser. 11. Chapman and 

Hall, Lond., 291 p. 

Reijnders, P. J. H. 19801. Organochlorine and heavy-metal residues in harbour seals frorrt the 

Wadden Sea and their possible effects on reproduction. Netherl. J. Sea Res. Y14:30G1. 

Reijnders, P. J. H. 1984. Man-induced environmental factors in relation to fertility clhanges in 

pinnipeds. Errviron. Consem. 11:6165. 

Reijnders, P. J. H. 1986. Reproductive failure in common seals feeding on fish from polluted coastal 

waters. Nature 324:456457. 

Tanabe, S., S. Wata:nabe, H. Kan, and R. Tatsukawa. 1988. Calpacity and mode of PCB metabolism 

in small cetaceans. Mar. Mamm. Sci. 4:103-124. 

Walker, C. H. 1983. Pesticides and birds--mechanisms of selective toxicity. A.gric. Ecosystems 

Environ. 9:211-226. 

Wilkinson, D. 1991. Report to Assistant Administrator for Fisheries: Prograrn Review of the 

Marine Mammal Standing Networks. U.S. Dep. Commer., NOAA, Nat. Mar. Fiish. ?em., 

Wash., D.C., 171 p.. 

Wise, S. A., B. J. Koster, and R. Zeisler. 1988. Status of specimen banking activities at the 

National Bureau of Standards. S. A. Wise, R. Zeisler, and G. M. lGo~ldstein (Editors), 

Progress in cenvironmental specimen banking. U.S. Dep. C:omrner., Nlat. Bur. Standards, 

Gaithersburg, Md, NBS Spec. Publ. 740, p. 10-18. 

Wise, S. A., B. J. Koster, IR:. M. I'arris, M. M. Schantz, S. R. Stone, and R. Zeisler. 1'989. E~cpericences 

in environmental specimen banking. Int. J. Environ. Anal. Chem. 37:91-106. 

Appendix I, National Marine Mammal Tissue Bank Protocolls 

Sampling Materials 

The foll~owing materials are used for collecting marine mammal tissue sample!+. 'These materials, 

except where noted, will Ibe provided by the NBSB: 

Teflon sheeting, 

Dry shippers (LN2 cooled) with shipping containers, 

Lhl2 in container for freezing samples on site, 

Dewar and lid, 

Lalb coats (disposablle), 

In~~ulated gloves, safety glasses, and tongs for handling the LN2 and frozen sannples, 

Balance for weighing, samples (supplied by the facility performing the necropsy), 

Surgical scissors, forceps (supplied by the facility perfoming the neciropsy), 

Lalbels for exterior of sample jars, 

Tape for swuning exterior jar labels, 

Lid labels, 

Shipping labels, 

Data recording, forms ( an example is shown on pages 33-32!). 

In addition, the coll~ection of two replicates of a single sample requires: 

4 - pairs non-talced vinyl gloves, 

1 - 14x1s Teflon FEP (Fluorinated ethylene propylene) bag or shwt to provide i3 clean 

working sul-face, 

1 - 12x12" Teflon FEI' bag for transporting sample from the field to the processing fac~ility, 

2 -Teflon jars (180 mlL, 49 mm diameter, 120 mrn length) with lid labels, 

2 -500-mL Teflon boittles containing high purity distilled or best available 'water for linsirrg 

samples, 

1 -Titanium bllade/'r'eflon TFE (Tetrafluoroethylene) handle knife, and high grade 

ethanol for rinsing knife (1 L). 

Sample Collection 

General 

Stage I, tissue remov,al, may occur under conditions of limited controll. 

Sttage 11, tissue processing, will occur under laboratory corlditions where possible; this includes 

shipboard labboratories. 

Stage 111, sample pankagirrg and shipping should be relatively standard for all tissues and 

should not vary. 
Stage I, Tissue Removall 

1. Basic Co~~siderations for all Sampling 

a. The size of the tissue sample removed from an animal should be sufficient to provide two 

sub!;amples of 150-20CI g each for archival. 

b. The anatomical location of the tissue removal is specified in order to maint,ain consistenc~y 

and comparab~ility between the same tissue types. 

c. Sterile, cleaned, non-talced vinyl gloves are used by all personnel involved in sample 

removal and handling. Calution must t~ taken throughout the procedures to1 reduce the risk 

of chemical co~ntamimltior~ of the sample. 

d. No animal will be considered a candidate for sampling, if tissues cannot be collected 

within the specified time after death ;and if handling of the animal has not folliowed 

prol.0~01 criteria or can not be documented. 

2. Procedures 

a. Record informcation on location, time taken, and other pertinent data on the individual 

anirnal on the data recording fonm. 

b. Takre and record all measu:remenks required on the data recording form[. 

c. Witlh the anim,al lying on its side, confirm gender. 

d. Using a stainless steel inst~rumenlt, make an incision through the skin and blubber on the1 

donil side, anterior to, the dorsal fin and posterior to the blow hole and measure blubber 

thickness. 

3. The tissue samples ,are removedl as soon as possible after opening the body cavity. Opening the 

body cavity and initial cutting of the skin to expose adipose and muscle tissue may be perfonmad 

with high quality stainless steel dissection tools previously rinsed in high purity water. 

a. Adilpose Tissure (Blubber) 

The anatomicall site of blulbber relmoval will depend on the distribution of f,at layers on the 

ani~nal. 

An incision is made anterior to the dorsal fin and posterior to the blow hole with a 

'stainless steel implement. Tvvo more cuts are made perpendicular to the first incision 

]near the fin and bllow hole respectively on the left side of the animal. A three-side 

flap of skin is cut. Using the titanium knife and grasping a comer of the blubber with 

hemostatic forceps, separate a section of blubber from the muscle layer which is below 

the blubber. After the blubber is separated from muscle, separate the blubber from the1 

skin in the same manner. A total of 300 g of blubber are required for storage. Place Ithe 

blubber in a clean Teflon bal3 for immediate transport to the processing area. 

b. Liver 

Not(. the general appearance of the liver before removal, including any unusual coloration, 

texture, shape, etc. 

Preferably using a stainless steel instrument, remove the entire lliver a~nd place it in a 

clean Teflon bag. If the enlire liver can be removed, use a titanium knife to remove a 
1X)-200 g section from both sides of the medial indentation at the distal end of the 

liver and place in a clean Teflon bag for immediate transport to the processing area. 

4. Following the removal of the NBSB samples, any additional samples and1 m~easurements are 

taken and tlhe informa~tion is included on the NBSB Sample Data1 Fom. 

Stage 11, Tissue Processing, 

1. Tissue processing should take place within a laboratory facility under the cleanest cc~ndi 1.i ons 

available. The processing area of the laboratory should be cleaned to remove dust and the 

working surfaces covered with Teflon sheeting. 

2. Onlly titanium knives are to be used to cut samples during Stage 11. 

3. Liver 

a. Tare the weight of the pre-cleaned Teflon jars, record that ,weight, and lalbel the jar for the 

sample. 

b. Remove the sam~ple from the Teflon bag. Holding the sample over a sink,, rinse the surface 

of the specimen with water from the Teflon bottle. Pour approximately '100 ml or more of 

the water from the Teflon bottle over the specimen to wash off blood and other fluid. Rub 

the specimen with gloved hand, if necessary, to remove bbod, etc. Allo~~ the specimen to 

drain for several minutes. Place the rinsed specimen on a dc?an Teflon sheet. 

C. If the whole liver is transported to the processing facility, remove a 150-200 g portion of 

the distal end of both sides of the medial indentation using a titanium1 knife. Dissect each 

150-200 g portion into two equal parts and place one section in the A jlar and the 'other 

section1 in the B ja~r. Repeat the procedure for the remaining 150-200 g section removed from 

the other distal end. Each sample must fit in a Teflon jar with a volume olf 180 rr~L (49 mm 

diameter and 120 rnrn length). 

d. If the liver sample is 300-400 g, use the titanium knife to divide the specimen into two 

equal samples (Sample A and Sample B) of 150-200 g each. Each sanlple must fit in a 

Teflon jar with a ,volume of 180 ml(49 mm diameter and 120 mm length:). 

e. After the sample has been dissected and placed in the Teflon jar, weigh the jar and record 

with the tared weight. 

f. When the work on the sample has been completed, rinse the titanium knife with the water 

and rub with the gloved fingers to remove all blood and fluids from the1 knife befort? they 

have time to dry. Rinse the knife with ethanol and air dry. 

g. Contir~ue to Stage 111, Tissue Packaging and Shipping.. 

4. Blubber 

a. Tare the weight of the pre-cleaned Teflon jar, record that and label the jar for the sample. 

b. Remove the specimen from the Teflon bag; pour approximately 100 ml or more of the wa ter 

from the Teflon bottle over the specimen to wash off bltmd and other fluid. Allow the 

specimen to drain for several minutes. 
c. Use the titanium knife to removle any remaining portions of muscle or skin attached to the 

blubber and to trim edges cut with the stainless steel implements. Also, any tissue tha~t 

came in contact with iI contamirlant, i.e., animal hair, hemostats, etc., should be excised 

and discarded. Excise a 300-400 1; section from the specimen. 

d. Using a clean titanium1 knife divide the specimen into two equal samples (Sample A and 

Sample B) of 150-200 g each. Each sample must fit in a Teflon jar with a volume of 180 ml, 

(49 mm diameter and 1'20 mrn length). 

e. After the sample has been dissected and placed in the Teflon jar, weigh the jar and record 

the tared weight. 

f. Aftel- the sample has been dissected angd placed in the Teflon jar rinse the titanium knif'le 

with the water and rub with the! glovied fingers to remove all blood and fluids from the 

knife before they have time to dry. Rinse the knife with ethanol and air dry. 

g. Continue to Stage 111, Tissue Packaging and Shipping. 

Stage 111, Tissue Packaging andl Shipping,, 

1. Record tl?e weights on the data recording form and sample labels. Affix the sample labels to 

the jars vvith wide tape and place the lid labels in the recessed slot on the jar lid. 

2. Freeze e<ach sample by immersing in the LN2 for 10 minutes. If LN2 is not immediatel:)~ 

available, freeze sample in freezer at: -70 OC: or lower. 

3. The LN2 shipper should be filled with liquid nitrogen for at least 6 hours to fully prepare it forr 

shipping. Pour off the excess LN2 anid place the frozen samples in the shipper (10-12 jars perr 

shipper). 

4. Once the shipper is full, transport it to the NIST; do not store the samples iln intermediate 

freezers. 

5. Double check the Data Recording Forms for completeness and accuracy. Any de~riatiomj or 

modificaitions of the protocol must be noted on the Form. 

6. Place a copy of the comp1et:ed forms in the shipper. 

7. Samples should be shipped1 within 413 hours or as soon as possible after sample collection using 

24 hour express package service to: 

Natio~nal Instibte of Stimdards and Technology 

Route 270 and Quince Olrchard Road 

Building 235, Room 8118 

Gaithersburg, Maryland 20899 Attn: Barbara Koster (301) 975-6291 

Shipping, expenses will be borne by NIST. Do not ship late in the week, i.e., Thursday, air 

Friday, or before holidays unless special arrangements have been made with the shipping 

service and NIST. 

8. The Specimen Bank persoinnel shou~ld be notified by telephone as soon as possible after the 

specimerls are shipped: Barbara Koslter (301) 975-6291, or Steve Wise (301) 975-31112. 
Sample Archival 

1. Samples are received at NIST and transported to the NBSB facility. 'The shippers are 

unpacked and sannples are inspected for any packaging problems and for lunsuitable 

temperatures. Data Recording Forms and samples are comparebd to insure thi~they correspond 

and that a111 informaition has been included. These samples are stored in a temporary LN2 

freezer anti are log@ into the temporary storage log book. They remain in 1:emporary siborage 

until assigned an NB!SB number and permanent LN2 freezer space. 

2. When the samples are moved into the permanent freezer lc)cation, a storage form, which 

contains storage location information, is completed, and the information is enterebd into the 

inventory form on the hlBSB personal computer. The samples are placed in cylindrical 

cardboard tubes (6.0 cm diameter x 63.5 cm length); each tube will hold up to five samples. 

These samples will remain stored in the LN2 freezer at about -150 "C until1 they are requested 

for analysis. 

3. Informatialn describing the animal is recorded and maintained, both in ha~rd copy and in a 

computer database, as part of the documentation for each sample in the Tissu~~t Bank. 

4. The duplicate samplt?s of each tissue (Samples A and B) are stored in different LN2 ]freezers to 

provide additional security. Sample A is intended for long-term storage vvhile sample B is 

available flor any analyses as required. 

5. As part of the specimen banking activities, NIST determines the concentrations of selected 

organic and inorganic constituents in approximately 10-20% of the archived specimer~s in order 

to (1) evaluate the stability of the specimens during long-term storage, (2) compare with data 

obtained firom other laboratories analyzing similar samples, which were collbected at the same 

time from the same !site as part of the monitoring component (i.e., quality assurance), and (3) 

compare with data fr~om samples to be collected in the future for monitoring lalng-ternn trends in 

pollution. An important aspect of these analyses is that they provide both inorganic and 

organic baseline data on the same specimens. Additional samples may be ijnalyzd by other 

laboratoric!~ as part of collaborative efforts with the NMMTl3. Samples to be analyzed are 

homogenin.ed using 'Teflon instruments and a cryogenic grinding procedure cies~igned to minimize 

sample cointamination and reduce the likelihood of changes in sample coimposition due to 

thawing and refreez.ing (Zeisler et al., 1983; Wise et al., 1989). 

Literature Cited 

* 

Wise, S. A., B. J. Koster, li. M. Parris, M. M. Schantz, S. R. Stone, and R. Zeisler. :1989. Experiences 

in environmental specimen banking. Int. J. Environ. Anal. Chelm. 37:91-105. 

Zeisler, R., S. H. Harrison, and S. A. Wise (Editors). 1983. The Pilot Nation~~al Environmental 

Specimen Bank -- analysis of human liver specimens. U.S. Dep. Cornmer., Nat. Bur. 

Xtandard~s, Gaithersburg, Md, NBS Spec. Publ. 656, 135 p. 
Data Recording Form 

1 I"' 1 I -- 

NATIONAL BlOMONlTORlNG SPECIMEN BANK 

Sampling Data 

N$ational Alarine Mammal Tissue Bank 

Anlmal ID Number --- Species 

Sample Sc~urCe ------- 

Collection Site ID Lat. Long. 

dav mo vr hr 

Time of D~eath: Method of Collectlon 

Intermedlai!e Storage (templrt?marks) 

Necropsy Location 

Sample Type: a Llver Kldney a Muscle a Blubber a Other --- 

,*a~ I mo , 1~ , b 

Time of Collection: Collected by 

Interrnedltile Storage (templrcmarks) --- -__- --- --_- 

[~BY, mo , 111 , Tlme of Preparation: 

Time of LN, Freezlng um 

llme Shipped From Slte: [ 1 1 1 

Tlme Recolved at Archlve: [m 

tl Processor ------.--- 

0 

Shlpper .______.- 

Recelver _________- 

Protocol: O~tandard O~odific!d (Please note modlflcatlon below) 

Remarks: 

page 1 of 3 
Animal ID Number 

Condition: • Alive Freshly &tad Other (explain) - ------- 

-- 

Sex: Cl Male a Female Total length -I Total weight [ 

-- 7 -- 

Baleenrrooth counts (erupted or total) UUL [I 

Specify Unit of Mea:suremenli's 

Snout to melon 

Snout to blow hole 

-- 

Snout to center of anus 

-- 

I Snout to center of genital aperture [ -- 7 -- -- 

snout to center of eye r 7 snout to ant. lnsenlon of flipper r 7 

--- 

snout to ant. insertion of fin I --- 1 Flipper length 

Snout to fin tip Flipper width 

--- 

Snout to fluke notch Fluke width 

Snout to caudal end Fin height 

of ventral grooves 

Girth: Axlliary I-1 Max (location) -1 

Blubber thickness (Iocathon) 

dorsal 1-1 lateral [ J -- 

ventral L_ 3 

Parasitces (location) 

internal -- -- 

Stomac:h contents - -- 

Reproductive condition: Preg.Lac:tatlnglFetus length 

Gonad ~t. m lenrl right 17 
Anlmal ID Nunlber-- 

Method 101 Collection -- ----- 

Age --- Method of Age E!stlmatlon 

General Comments: 

General Appearance of Indlvldual: 

General Appearance of Organs: 

Hlstologlcal Samples: 

Indlvlduali'Organlzation - --- 

Final Destlnatlon ---. 

Tissues S'ampled ---. 

Sample Welgh1:s: A B 

Llvc?r 9. 11. Blubber 

Kldney 

Prepared by: 

Name (print) 

Other 

A copy of thls form ahould be 

shlpped wllh samples lo: 

Nallonal lnsdtute of Standards ancl Techno1og.y 

Bldg 235. Rm 0118 

Galthersburg, MD 20899 

Attn: Barbara Kosler 

(301) 975-6291 

page :1 of 3 
Appe~lldix 11, National Marine Mammal Il'issut: Blank 

Access Policy 

The goal of the Na~tional Marine Mammal Tissue Bank (NMMTB) is t:o maintain quality 

controlled marine nnammal tissues to permit retrospective analyses for the purposie of, 

determining enviror~mental trends, and conducting analyses using new and innovative 

analytical techniques. 

The National Marine Fisheries Service (NMFS), Office of Protected Resources (PR) has the 

responsibility for managing the NMMTB. NMMTB tissues are bankled in the National 

Institute of Standards and Technology (NTST) National Biomorutoring Specirnen Bank (NBSB) 

located in Gaithersburg, Maryland. 

Duplicate samples (:denoted A and B) of -150 g for each tissue specimen are banked in the 

NMMTB. Subsamples of the "B" samples can be homogenized ancl aliquoted into 

approximately 20 subsamples of 6-8 g each which are then available for scientific research. 

The "A samples will remain as bulk samples and will only be homogenized after all portions 

from the "B" sample are depleted and sufficient justification exists to homogenize the 

remaining material. Thus, 50% of each specimen is available to the scientific comnnunity for 

research and scientific evaluations consistent with the goals of the NMMTB 1:)rograrn and 50% 

is intendedl for long-term storage as a more permanent archive for pasterity. 

The homogenized subsamples of "B samples are divided into three categories. C'ategtory 1 

constitutes 40% of the homogenized material and is available to the scientific comnnunily for 

research that is con!astent with the goals of the NMMTB. Formal requests for access to these 

tissues must be madle as described below. Category 2 consists of 50% of the homogenized 

material and is reserved for use by the contributing agencies. A procedure for access to these 

samples will be established by the contributing agencies and lhe NMMTI3. Category 3 ]is the 

remaining 10% of the homogenized material which is reserved to perform baseline analyses of 

the specimens to monitor storage stability, to compare with analyses from real-time monitoring 

programs imd to corkduct research investigations within the NMMTB program 

If the "A" sample is eventually homogenized, the subsamples are divided into the following 

four categories. Category 1 consists of 25% of the material aind is available to the scientific 

conununit.y as described above. Category 2 consists of 25% of the material and is reserved for 

use by the contributing agencies. Category 3 consists of 20% of the material Ibr use within the 

NMMTB program as described above. Category 4 is the wemaiining 40% which is intended as a 

permanent archive Ithat will not be utilized unless high priority needs exist; ithe dett!rmination 

of this need will be made by an advisory committee to the NMMTB. Cornbuning &re "A" and 

"B samples, the total percentage sample allocations are as follows: Category 1 (3;!.5%), 

Category :2 (37.5%), Category 3 (lo%), and Category 4 = (20%). For clarification, the relative 

amounts alf each category are illustrated in the following figure. 

Formal requests for banked tissues in Category 1 must be subm~~tted to the Director of the NMFS 

Office of Protected Resources. The request will be reviewed b,y an informal review committee 

consisting of no less than three individuals, who are members of the NPVIMTB's Team of 

Scientists. The review committee will be convened by the Director and shall have 

representation from both the government and academic/private sector. At least one member of 

the review commitltee shall be an expert in the field that is rielat& to the proposed research 

activity. The Director will make the final decision based on the advice providled by the 

review committee. 
Scientific 

Community 

Contributing 

Agency 

--, 

J 

Scientific 

Community 

r 

\ 

Contributing 

Agency 

Request:; for sample:^ from the NMMTB must include a clear and concise statement of the 

proposed work and be consistent with the goals of the NMMTB. The folllowirlg specifi~c 

information should be included in the request for samples: 

11. 

Z! . 

:I. 

4. 

5. 

ti. 

7. 

8. 

$1. 

Name of principal investigator and affiliated research or academic- organization, 

Specific tissue sample and quantity desired, 

Explanation of proposed research to be conducted, 

Justification for use of banked tissue, 

Research facility where analyses will be conducted, 

Analytical quallity control procedures to be used, 

Estimated completion of research, and schedule/date of subsequent reports, 

Agreement that all results/findings be reported to the NMMT'B, 

Agreement that credit and acknowledgement will be given to NOAA/MFS and 

the NMM:TB for use of banked tissues. 

Of particular importance in determining whether a sample request will be granted is the 

justification that samples from the hIMM'T'B are required to accomplish the relsearch and that 

suitable samples to accomplish the goals of the proposed research could not be obtained fro113 

other sources. The NMMTB is not intended to be used as a readily accessible source of marinlce 

mammal tissue for any researcher needing specimens, but only for research requiring banked 

samples from the past. 

An informal review process will also be set-up for banked tissues withdrawn from Category 2:; 

however, the final decision on Category 2 samples will be made by the contributing agelnqr. 

Use of Category 3 tissues will be determined by NMFS and NIST. Category 4 tissues cannot be 

withdrawn until all tissues from the other categories have been depleted. When Category 4 

tissues are withdrawn, the same re~niew process will be followed as described for Category 1 

tissues with the final determination made by the Director. 
Formal requests shall be submitted to the following address: 

Director, 

Office of Protected ItYesources (F/PR) 

National Marine Fisheries Service, NOAA 

US. Department of Commerce 

1335' East-West Highway 

Silver Spring, Md :20910.