NOAA Technical Memorandum NMFS-PIFSC-2

May 2004

Contingency Plan for Hawaiian Monk Seal 

Unusual Mortality Events 

Pamela K. Yochem, Robert C. Braun, Bradley Ryon, 

Jason D. Baker, and George A. Antonelis 

Pacific Islands Fisheries Science Center 

National Marine Fisheries Service 

National Oceanic and Atmospheric Administration 

U.S. Department of Commerce 
About this document

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Author. Date. Title. U.S. Dep. Commer., NOAA Tech. Memo., NOAA-TMNMFS-PIFSC-XX, xx p. 

__________________________ 

For further information direct inquiries to

Chief, Scientific Information Services 

Pacific Islands Fisheries Science Center 

National Marine Fisheries Service 

National Oceanic and Atmospheric Administration 

U.S. Department of Commerce 

2570 Dole Street 

Honolulu, Hawaii 96822-2396 

Phone: 808-983-5386 

Fax: 808-983-2902 

___________________________________________________________ 

Cover photo courtesy of Brenda L. Becker, Pacific Islands Fisheries Science Center 
Pacific Islands Fisheries Science Center 

National Marine Fisheries Service 

National Oceanic and Atmospheric Administration 

U.S. Department of Commerce

Contingency Plan for Hawaiian Monk Seal 

Unusual Mortality Events 

Pamela K. Yochem,1

 Robert C. Braun,2 Bradley Ryon,3

Jason D. Baker,2

 and George A. Antonelis2

1

Hubbs-Sea World Research Institute 

2595 Ingraham Street 

San Diego, California 92109 

2

Pacific Islands Fisheries Science Center 

2570 Dole Street 

Honolulu, Hawaii 96822-2396 

3

Pacific Islands Regional Office 

1601 Kapiolani Boulevard, Suite 1110 

Honolulu, Hawaii 96814-4700 

NOAA Technical Memorandum NMFS-PIFSC-2

May 2004

 iii 

CONTENTS 

Contents Page 

1. Background 1 

1.1 Distribution and Habitat Use 1 

1.2 Population Biology 1 

1.3 Mortality Patterns 3 

1.4 Unusual Mortality: Past Events 5 

1.4.1 Hawaiian Monk Seal Unusual Mortality Event, 

Laysan Island, 1978 5 

1.4.2 Hawaiian Monk Seal Unusual Mortality Event, 

Laysan Island, 2000-2001 5 

1.5 Background of this Document 6 

 

2. Federal/State Authority and Jurisdiction 6 

 

3. The On-Site Coordinator 7 

3.1 The Appointment of the On-Site Coordinator 7 

3.2 Responsibilities of the Federal On-Site Coordinator 8 

4. Pre-Event Planning for Rapid Response 8 

4.1 Building a Response Team 8 

4.2 The Administrative Support Team 9 

4.3 Scientific Advisors 9 

4.4 Training and Readiness 9 

4.5 Establishing MOUs with Cooperators 10 

4.6 Pre-Planning for Sample Analyses 10 

5. Recognizing an Unusual Event 10 

5.1 Guidelines for a Graduated Response 11 

5.2 Protocol for Initiating Unusual Event Designation 12 

 

6. Notification of Personnel and Agencies 12 

 

7. A Tailored Response 12 

7.1 Objectives and General Guidelines 12 

7.2 Responding to an Event of Known Cause 13 

7.3 Investigating an Event of Unknown Cause 14 

7.4 Determining “The End” 14 

7.5 Post-Event Assessment and Monitoring 15 

8. Communication and Reporting 15 

9. Public Health Concerns 15 
 iv 

(CONTENTS, continued) 

Page 

10. Resources: Location and Utilization 16 

10.1 Introduction 16 

10.2 Equipment and Logistics 16 

10.3 Information and Data Resources 16 

10.4 Development of Temporary Animal Housing and 

Emergency Facilities 17 

10.5 Funding 17 

11. Response and Investigation Protocols 17 

11.1 Rescue and Rehabilitation 17 

11.2 Release of Rehabilitated Monk Seals after a MMUME 17 

11.3 Procedures for Carcasses 17 

11.4 Tissue Sample Collection 17 

11.5 Environmental Assessment and Sampling 18 

11.6 Packaging and Shipping Samples 18 

11.7 Keeping Records and Maintaining Chain of Custody 18 

11.8 Surveys 18 

 

12. Acknowledgments 18 

13. References 19 

14. List of Appendices 23 

 

A: Hawaiian Monk Seal Specimen Collection A-1 

B: Hawaiian Monk Seal Necropsy Protocol B-1 

C: Equipment and Supplies for a Single Necropsy C-1 

D: Hawaiian Monk Seal Rescue and Rehabilitation Guidelines D-1 

E-1: Additional Samples Requested during the Laysan 2001 MMUME E-1 

E-2: Specimen Collection Protocols for Specific Biotoxins E-3 

E-3: Specimen Collection Protocols for Chemical Pollutants E-7 

F-1: Epidemiology Sampling Supply List F-1 

F-2: Veterinary Medical Kit F-5 

G-1: Monk Seal Specimen Collection Summary Form G-1 

G-2: Hawaiian Monk Seal Tagging & Chemical Immobilization Form G-3 

G-3: Hawaiian Monk Seal Clinical Examination Form G-7 

G-4: Hawaiian Monk Seal Field Hematology Report G-11 

G-5: NMFS Human Interaction Forms G-15 

G-6: Marine Forensics Chain of Custody Form G-19 

H-1: Federal, State and Local Authorities H-1 

H-2: Potential Response Team Members H-3 

H-3: Diagnostic Laboratories H-7 

H-4: Cooperating Experts H-11
 v 

(CONTENTS, continued) 

 Page 

I-1: Flowchart and timing of response to MMUME in the United States I-1 

I-2: Flow Diagram for Suspected Marine Biotoxin Incidents I-3 

I-3: Flowchart for response to a marine mammal mortality event with 

known vs. unknown cause I-7 

J-1: MOU with participants in UME response J-1 

J-2: MOU with laboratories/individuals performing tests/analyses J-3 

K-1: Excerpts, 28 August 2001 Update of the Hawaii Area 

 Contingency Plan K-1 

K-2: Excerpts, Oil Spill Field Operations Guide K-5 
 
 

1. Background 

1.1 Distribution and Habitat Use 

 Hawaiian monk seals (Monachus schauinslandi) were designated as depleted under 

the Marine Mammal Protection Act and endangered under the Endangered Species Act in 

1976, following declines of 50% from the late 1950s. In 1988, critical habitat for monk 

seals was designated as the emergent land, lagoon waters, and ocean waters out to a depth 

of 20 fathoms (37 m) around breeding islands (excluding the main Hawaiian Islands) and 

Maro Reef. In 1991, a Protected Species Zone was established out to 50 nautical miles 

from the islands and the corridors between islands. 

 There are six main reproductive populations of Hawaiian monk seals in the 

Northwestern Hawaiian Islands (NWHI): French Frigate Shoals (FFS), Laysan Island, 

Lisianski Island, Pearl and Hermes Reef, Midway Atoll, and Kure Atoll (Fig. 1). Smaller 

numbers are present on Necker Island and Nihoa Island and on the main Hawaiian 

Islands. Monk seals are rarely observed at Johnston Atoll where one female pup was born 

in 1969. 

 Approximately 90% of Hawaiian monk seals remain at their natal site for life; the 

remaining 10% move between or among major population centers. Studies conducted at 

FFS in 1996 and 1997 documented the use of foraging habitats by monk seals which 

were much farther from breeding locations than was previously known or anticipated. 

Similar studies in 1997-98 at Pearl and Hermes Reef documented very little extra-atoll 

movement. Most feeding appears to occur at depths less than 75-90 m, though seals 

occasionally dive to depths exceeding 500 m. Known prey items include reef fishes, 

benthic fishes, cephalopods, and crustaceans. 

1.2 Population Biology 

Since 1985, the average rate of decline was approximately 3% yr-1, although the 

beach counts1

 have been stable from 1993 to 2000 and declined again in 2001 and 2002 

(NMFS, unpublished data). Further decline is likely due to high juvenile mortality and an 

inverted age structure at FFS, the largest colony (Carretta et al., 2001). The annual 

number of births has varied substantially over the past decade and is expected to decline 

in the near future due to poor recruitment at FFS. At present the species numbers 

approximately 1400 and is the most endangered species of marine mammal that lives 

entirely within U.S. jurisdiction (Carretta et al., 2002). 

 

1

 Direct enumeration data cannot be used for characterizing long-term trends because sufficient 

field investigation in the NWHI has not been consistently undertaken at all sites and years. 

Instead, a measure of long-term trend is derived from the mean of all the beach counts that have 

been conducted with varying frequency since the late 1950s. Beach counts provide a useful index 

of population trends and do not account for seals at sea. Approximately 1/3 of each seal 

population is on land during a survey, but the exact proportion varies (e.g., by site and season). 
 2 

Fig. 1. The Hawaiian Archipelago, with the six main monk seal breeding and pupping 

sites identified (French Frigate Shoals, Laysan I., Lisianski I., Pearl and Hermes Reef, 

Midway Atoll, Kure Atoll). 

 Females give birth on beaches with adjoining shallow waters, which provide 

protection from sharks. Most pups are born between February and August, with a peak 

from late March to May (Johnson and Johnson, 1980; Johanos et al., 1994). Females give 

birth for the first time at ages 5 to 10, and typically 50%-70% of all adult-size females 

give birth each year (Johanos et al., 1994). Gestation is approximately 1 year, with a 

mean interval between births (for those females that pup in consecutive years) of 381 

days (Johanos et al., 1994). Females fast during the 5 or 6-week nursing period, and 

nursing pups are sometimes exchanged between females (Kenyon and Rice, 1959; Alcorn 

and Henderson, 1984; Boness, 1990; Gerrodette et al., 1992). Mating occurs 3 or 4 weeks 

after pups are weaned. Although mating takes place in the water and is rarely observed, a 

mating peak in May and June is inferred from the occurrence of fresh mounting injuries 

and association patterns of males and females (Johanos et al., 1994). Unlike other 

gregarious pinnipeds, monk seals are typically solitary and do not form large, seasonal 

breeding aggregations. They have a serially monogamous social system with males 

forming a dominance hierarchy. 

 Pups are born with a black coat, which is shed gradually at the end of the nursing 

period and replaced by pelage that is silver gray on the dorsum and sides and beige on the 

venter. Juveniles and adults undergo a “catastrophic” annual molt, in which the epidermis 

180° 175W 170W 165W 160W 155W

15N

20N

25N

30N

Nihoa I.

Necker I.

French Frigate Shoals

Laysan I.

Lisianski I.

Pearl and Hermes Reef

Midway Atoll

Kure Atoll

Northwestern Hawaiian Islands Main Hawaiian Islands

Johnston Atoll 
 3 

is shed with the hair in large patches (Kenyon and Rice, 1959; Yochem and Stewart, 

2001). The freshly molted pelage is silver gray above and beige below except in adult 

males, where it is dark brown. Molting peaks in July, and females with pups molt 5 or 6 

weeks after mating (Johanos et al., 1994). 

1.3 Mortality Patterns 

 Compared to other marine mammals, very good information exists on mortality rates 

of Hawaiian monk seals. These rates are obtained annually by resighting known-age 

animals during 2-5-month field seasons at all six main subpopulations in the NWHI (Fig. 

1). Current tag and resight efforts are sufficient to obtain cohort-and age-specific survival 

(the complement of mortality) rates using Jolly-Seber analysis. These data are analyzed 

annually, and estimated rates are back-corrected when animals not seen for one or more 

years are subsequently resighted. Thus, long-term trends and variability in monk seal 

mortality rates are well characterized to age 15 years or more. These rates have reflected 

trends that greatly influence monk seal population trends. For example, juvenile mortality 

rates were low at most sites during the mid-to-late 1980s. Subsequently, juvenile 

mortality increased throughout the archipelago to varying degrees and on different time 

frames. Most notably, very high juvenile mortality has led to a severe and enduring 

population decline at FFS. Recent decline in juvenile survival has also been reported at 

Pearl and Hermes Reef and Midway and Kure Atolls in 2001 and 2002 (NMFS 

unpublished data). 

 Although annual mortality rates are well characterized, the timing and relative 

importance of various causes of mortality are not well known. Jolly-Seber rates are 

available only at the end of a field season, so these data cannot be used to evaluate 

mortality processes in real time. Moreover, as noted above, these rates are annually 

updated and back-corrected with each new year’s data. The exact timing of mortalities is 

rarely known, as field effort encompasses less than half of any given calendar year. Even 

within a field season it is rarely possible to determine when an animal has died because 

so few carcasses are observed on the beach. In the few cases where carcasses are 

available for examination, unequivocal cause of death is often not discernible. For 

animals that are no longer sighted during regular beach walks, one can only infer that 

they died some time after the final sighting. For the Jolly-Seber analysis, any animal 

sighted within the calendar year, whether the first day or the last, is scored as having 

survived to that year. This is necessary for calculation of annual rates but does not 

provide within-season information on time of death. 

 To address the information gaps noted above, field teams from the Hawaiian Monk 

Seal Assessment Program provide a weekly report of the cumulative number of 1- to 3-

year-old seals sighted to date, as well as the total number of confirmed and probable 

deaths within the season. In this way, minimum juvenile survival is regularly assessed so 

that unusually high mortality within a field season is likely to be detected. Post-mortem 

exams are performed, and samples are collected for evaluation by a team of marine 

mammal health and disease experts to determine healthy body condition and cause of 

death where possible. Some laboratory analyses (e.g., erythrogram and leukogram 
 4 

evaluations) are performed in the field; these preliminary findings, as well as descriptions 

of gross lesions or clinical signs observed, are reported to the Monk Seal Assessment 

Program and Monk Seal Health and Disease Program. These measures increase the 

likelihood of timely detection of an unusual mortality event, allowing a response to be 

mounted within the field season if deemed appropriate. 

 Known causes of mortality in Hawaiian monk seals include emaciation of juveniles, 

tiger and Galapagos shark attacks, male aggression (individual and multiple), deleterious 

fisheries interactions, and entanglement in marine debris (Balazs and Whittow, 1979; 

Kenyon, 1981; Henderson, 1985, 1990; Alcorn and Kam, 1986; Banish and Gilmartin, 

1992; Hiruki et al., 1993a, 1993b; Nitta and Henderson, 1993; Starfield et al., 1995). 

Disturbance of pregnant or nursing females likely causes them to desert preferred 

pupping beaches, resulting in decreased pup survival (Kenyon, 1972; Kenyon, 1981). 

Maximum age is believed to be 25-30 years, but few seals live this long (NMFS, 

unpublished data). 

 The influence of disease on monk seal populations is not well understood, but is an 

active area of research (Aguirre et al., 1999; Aguirre, 2000; Appendix A, Hawaiian Monk 

Seal Specimen Collection Protocol and Appendix B, Hawaiian Monk Seal Necropsy 

Protocol). Infectious and noninfectious diseases have been reported in Hawaiian monk 

seals but do not appear to be impeding population recovery: all wild seals carry parasites; 

dental and skeletal abscesses and pathology were the apparent causes of death in one 

aged seal; and biotoxins may pose a serious risk (Golvan, 1959; Rausch, 1969; Kenyon 

and Rauzon, 1977 [cited in Kenyon 1981]; DeLong, 1978; DeLong and Gilmartin, 1979; 

Gilmartin et al., 1980; Whittow et al., 1980; Dailey et al., 1988). Of the 15 Marine 

Mammal Unusual Mortality Events (UMEs) occurring in other marine mammals species 

since 1992 (Dierauf and Gulland, 2001: 72-73, Table 1), infectious diseases and biotoxins 

were the most common diagnoses (five cases each). Other factors implicated in UMEs 

included fisheries interactions, ship strikes, and large-scale decadal changes in oceanic 

productivity (Polovina et al., 1995). 

 

 In a recent review of emerging and resurging diseases in marine mammals, Miller et 

al. (2001) describes three ways in which wildlife species may be exposed to emerging 

diseases (newly evolved or newly identified as pathogens), all three of which are 

applicable to Hawaiian monk seals. The first is exposure via spillover from domestic 

species or humans. Many recent reports of transmissible diseases in marine mammals 

have included organisms traditionally associated with domestic animals and humans 

(e.g.; Brucella sp., Ewalte et al., 1994; Ross et al., 1994; Jepson et al., 1994; 

Mycobacterium bovis and M. tuberculosis, Forshaw and Phelps, 1991; Cousins et al., 

1993; Woods et al.; 1995; Bernardelli et al., 1996). The second method of exposure is via 

rehabilitation or translocation of animals (e.g., Stamper et al., 1998, reports the possible 

spread of leptospirosis from skunks to rehabilitating harbor seals). The third way that 

wildlife species may become exposed to emerging diseases is through the effects of 

environmental phenomena such as El Niño and large-scale climate change on the 

proliferation or spread of infectious organisms (Fauquier et al. 1998; Harvell et al., 1999; 

Reddy et al. 2001). Another source of exposure to infectious diseases in Hawaiian monk 
 5 

seals is other wildlife species. Morbillivirus has been detected in the North Pacific Ocean 

(in common dolphins; Reidarson et al, 1998), and cross-species infection has been 

described for this group of viruses (e.g., canine distemper in Baikal and Caspian seals; 

Mamaev et al., 1995; Forsyth et al., 1998). A virus closely related to dolphin 

morbillivirus was implicated in an UME involving Mediterranean monk seals (Osterhaus 

et al., 1998), although saxitoxins were also detected in these seals. 

 

1.4 Unusual Mortality: Past Events 

1.4.1 Hawaiian Monk Seal UME, Laysan Island, 1978. 

 At least 50 monk seals died at Laysan Island in 1978, primarily in juvenile-young 

adult (1-5 years) and older adult (18-30 years) age classes (Johnson and Johnson, 1981; 

Gilmartin et al., 1980). Clinical signs noted before death included gradual weight loss 

(over a period of weeks to months), lethargy, change in behavior patterns, poor quality 

pelage (particularly around the face and neck), and presence of fresh lesions (primarily on 

the rear flippers and lower back). Affected seals were emaciated and gastric ulceration 

was observed in all 15 dead seals for which complete tissue sets were collected; this was 

accompanied by frank hemorrhage secondary to heavy gastric nematode infestations in 

many seals. High levels of ciguatoxin and maitotoxin, neurotoxins produced by benthic 

dinoflagellates in association with tropical reef environments, were estimated by bioassay 

analyses of the livers of two seals examined; levels were 30-50 times higher than those 

found in the liver of a monk seal that had been maintained in captivity for 15 years. 

However, radioimmunoassays for ciguatoxin were not consistent with the bioassay 

results: one seal had liver ciguatoxin levels 25% above those in the captive seal, and the 

second had levels 9% below the captive seal. Two of 18 live seals sampled had elevated 

white blood cell counts. One of these seals disappeared within 2 weeks of sampling, but 

the other remained healthy at least through 1979. 

1.4.2 Hawaiian Monk Seal UME, Laysan Island 2000-2001. 

 An Unusual Mortality Event was declared in the spring of 2001 following the death of 

four juvenile monk seals in a 9-day period at Laysan Island. Criteria used by the Working 

Group on Unusual Marine Mammal Mortality Events (WGUMMME) included increased 

mortality compared to prior records, mortalities occurring in a localized area, mortalities 

associated with abnormal behavior patterns, and the mortality of a critically endangered 

species (Wilkinson, 1996; Antonelis et al., 2001). An investigation was launched at 

Laysan Island and other subpopulations in the NWHI to assess possible rangewide 

effects. Survival from weaning to 1 year in 2001 was lower than most previous years at 

all but one site. Investigation into causes included infectious diseases, parasitism, 

environmental conditions, naturally occurring biotoxins, and anthropogenic 

contaminants. Necropsy results from 11 seals revealed emaciation but no signs of 

infectious disease or toxicosis linking the UME deaths. The unusually high mortality in 

yearling seals was not correlated with their size at weaning, but appeared to be a result of 

their inability to forage successfully during the post-weaning transition to nutritional 
 6 

independence (Antonelis et al., 2001). High juvenile mortality was also observed in 2002 

(NMFS unpublished data). 

1.5 Background of this Document 

 The first attempt to develop a response plan for UMEs in Hawaiian monk seals is 

presented by Gilmartin (1987). The National Marine Fisheries Service (NMFS) has 

published a National Contingency Plan for responding to unusual marine mammal 

mortality events (Wilkinson, 1996). The WGUMMME recommended that a separate 

plan, consistent with the national plan and other relevant documents such as the Hawaii 

Area Oil Spill Contingency Plan, be prepared for Hawaiian monk seals because of their 

endangered status and the logistic difficulties associated with the remoteness of most of 

their colonies. A separate plan was prepared for manatees by Geraci and Lounsbury 

(1997), and the Hawaiian monk seal contingency plan follows the manatee format. 

2. Federal/State Authority and Jurisdiction 

 The Marine Mammal Health and Stranding Response Act was passed by the U.S. 

Congress in 1992 (P.L. 102-587, 16 U.S.C. 1421 [c-d]), and it became Title IV of the 

Marine Mammal Protection Act (MMPA). Section 404 of MMPA Title 4 (P.L. 103-238, 

§24(b)(1)) established a framework for responding to Marine Mammal UMEs, described 

in detail in the National Contingency Plan for Response to Unusual Marine Mammal 

Mortality Events (Wilkinson, 1996). The response sequence is outlined below and is 

presented as a flowchart in Appendix I-1. 

• An increase in stranding rates or unusual findings is detected through one of three 

likely mechanisms:

o NMFS Regional Stranding Coordinator (Pacific Islands Regional Office, 

PIRO) detects increased strandings or unusual findings upon reviewing 

Level A data and consulting with the Stranding Network;

o Stranding Network directly notifies the NMFS Regional Stranding 

Coordinator (PIRO) of an increase in strandings or unusual findings;

o Monk Seal Research Leader observes an increase in strandings or unusual 

finding and notifies the Leader of the Pacific Island Fisheries Science 

Center’s Marine Mammal Research Program (MMRP). 

• The NMFS Regional Stranding Coordinator (PIRO) or the Leader of MMRP will 

contact the Executive Secretary for the WGUMMME (NMFS Headquarters), pass 

on the information, and request a consultation with the WGUMMME. 

• The Executive Secretary for the WGUMMME will formally request a 

consultation and forward a complete summary of the event in question and a 

historical record of stranding data to the WGUMMME. 
 7 

• Over a 24-hour period, the WGUMMME will review the data and compare it 

against the seven criteria for determination of an UME (Wilkinson, 1996:18): 

o A marked increase in the magnitude of strandings is occurring when 

compared with prior records. There is no set formula for determining what 

magnitude would trigger a response. The NMFS Southeast Region has 

used a formula of the historic mean plus two times the standard deviation 

to determine a threshold level. The WGUMMME stated that the 

magnitude must be weighed against other knowledge. As a pragmatic 

method, it was suggested that if a pulse in strandings is spread over an 

area or timeframe that strains the capacity of the Stranding Networks to 

respond, it should be cause for concern. 

o Animals are stranding at a time of the year when strandings are unusual. 

o An increase in strandings is occurring in a very localized area (possibly 

suggesting a localized problem), is occurring throughout the geographical 

range of the species/population, or spreads geographically with time. 

o The species, age, or sex composition of the stranded animals is different 

than those of animals that normally strand in the area at that time of year. 

o Stranded animals exhibit similar or unusual pathologic findings, or the 

general physical condition (e.g., blubber thickness) of stranded animals is 

different from what is normally seen. 

o Mortality is accompanied by behavior patterns observed among living 

individuals in the wild that are unusual, such as occurrence in habitats 

normally avoided or abnormal patterns of swimming and diving. 

o Critically endangered species are stranding. Stranding of three or four 

right whales, for example, may be cause for great concern whereas 

stranding of a similar number of fin whales may not. 

• The WGUMMME will return its decision to the Executive Secretary, and an 

official determination will be made if a quorum has been reached (i.e., majority of 

2/3 of the voting body). 

• The Executive Secretary officially requests concurrence from the NMFS Assistant 

Administrator. With this concurrence, the NMFS Assistant Administrator is also 

requested to appoint an On-Site Coordinator (see below). 

3. The On-Site Coordinator 

3.1 The Appointment of the On-Site Coordinator. 
 8 

 Following the sequence of events described above, the NMFS Assistant Administrator 

will appoint an On-Site Coordinator for the UME. The On-Site Coordinator is the 

appropriate NMFS Regional Administrator or his or her designee (Wilkinson, 1996). The 

selection of On-Site Coordinator is based upon recommendations from the WGUMMME 

and the Leader of the MMRP, PIFSC, NOAA Fisheries. 

3.2. Responsibilities of the Federal On-Site Coordinator. 

 The On-Site Coordinator should have strong management and leadership abilities, 

strong communication skills, the ability to make decisions with minimal use of 

intermediaries, the ability to access information and expertise (including interagency 

expertise), familiarity with the National Contingency Plan and associated protocols 

(including this monk seal contingency plan), and familiarity with and the ability to work 

with the Marine Mammal Stranding Networks (Wilkinson, 1996:19). The On-Site 

Coordinator must hold or be included on a federal Enhancement and Scientific Research 

Permit (e.g., the permit held by the Leader of MMRP). 

 The On-Site Coordinator will work closely with the Executive Secretary, NMFS 

national and regional offices (e.g., PIRO) and the Leader of MMRP. The On-Site 

Coordinator also will notify other federal agencies (e.g., U.S. Coast Guard, U.S. Fish and 

Wildlife Service), State of Hawaii wildlife resource agencies (e.g., Hawaii Department of 

Land and Natural Resources, Division of Aquatic Resources, and Division of Forestry 

and Wildlife,), Hawaii regional stranding network members, and the Hawaiian Monk 

Seal Recovery Team. A list of federal and state agencies and individuals to be notified is 

included in Appendix H-1. Contact lists are updated regularly (at least annually) by 

MMRP. Additional notifications will be made as needed; for example, the Hawaii 

Department of Public Health would be notified if a serious human health hazard (e.g., if a 

serious zoonotic disease is identified or suspected) existed on one of the main Hawaiian 

Islands, and USDA/APHIS would be notified if monk seal transport were being 

considered. The On-Site Coordinator will also assemble a response team as described 

below. 

4. Pre-Event Planning for Rapid Response 

4.1 Building a Response Team 

 

 The response team assembled by the On-Site Coordinator will include one or more 

wildlife biologists with knowledge of normal Hawaiian monk seal behavior and natural 

history and one or more veterinarians familiar with pinniped diseases and monk seal 

sampling protocols (including specimen collection and necropsy protocols). Additional 

team members will be selected depending on the size and nature of the event and may 

include a veterinary technician to assist with sample collection and processing, a rescue 

and salvage logistics coordinator, a parasitologist, and an environmental sampling 

coordinator. A list of potential team members is included in H-2; this list is updated at 

least annually by MMRP to ensure that a qualified team can be assembled immediately. 
 9 

4.2 The Administrative Support Team 

 Three important administrative areas to be addressed by the On-Site Coordinator in 

the event of a Hawaiian monk seal UME are financial administration, public information 

coordination, and tracking of animals and samples. As stipulated in the National 

Contingency Plan (Wilkinson, 1996), a certified Contracting Officer’s Technical 

Representative will be appointed as financial officer by the Regional Office and will 

work with the On-Site Coordinator and a certified Contracting Officer to negotiate and 

enter into agreements for services performed in the course of a MMUME response, to 

locate and order supplies and equipment, and to be responsible for prompt payment for 

services, purchase orders, and expense reimbursements. The roles of On-Site Coordinator 

and Contracting Officer’s Technical Representative can be filled by the same individual 

or by two separate individuals. The On-Site Coordinator will appoint a local media 

contact (either an agency public affairs officer or an individual involved in the response) 

to address public concerns and inquiries and to keep the NMFS Office of Public Affairs 

(301-713-2370) informed. Procedures for tracking data and samples will follow wellestablished protocols (Monk Seal Specimen Collection and Necropsy protocols, 

Appendices A and B). In some cases (e.g., potential litigation or prosecution related to 

the MMUME) chain of custody will be maintained for all samples and data collected 

during the response (Appendix G-6).

4.3 Scientific Advisors 

 In addition to the WGUMMME, the On-Site Coordinator may call on scientific 

advisors from the Hawaiian Monk Seal Recovery Team, the Marine Mammal 

Commission, or the list of attendees to the 11-12 September 2000 Hawaiian Monk Seal 

Health Studies Workshop. Additional advisors may be consulted depending on the size 

and nature of the event (e.g., the NOAA Marine Biotoxins Program, 843-762-8500, will 

be contacted if a Harmful Algal Bloom is suspected).

4.4 Training and Readiness 

 

 MMRP conducts extensive annual training of field personnel for the monk seal 

assessment program. Many of the topics addressed in this training program are relevant to 

MMUME response, such as the Monk Seal Health and Disease Program (including 

procedures and supply lists for specimen collection, necropsies, and field laboratory 

assays (Appendices A through H), field camp logistics and set-up, animal care and use 

policies, field communications and safety, and first aid training. In addition to this predeployment training, field camp personnel have practical experience in Hawaiian monk 

seal data collection and processing obtained during deployments of 2-5 months in the 

NWHI. 

 Monk seal rescue and rehabilitation are addressed in the Monk Seal Rehabilitation 

Manual (Appendix D). Lavigne (1999) reviewed past attempts to enhance monk seal 

survival and reproduction via the “headstart program,” the rescue, rehabilitation and 

transport of undersized females and the translocation or removal of adult males to reduce 
 10 

mobbing. Such efforts have had varying levels of success in enhancing monk seal 

survival. 

 The Leader of MMRP (Dr. George Antonelis, 808-983-5710) has designated 

individuals for key roles such as monk seal population assessment (Jason Baker, 808-

983-5711) and health studies (Dr. Robert Braun, 808-254-8181 or 808-254-3530 or 808-

783-6565; Lizabeth Kashinsky, 808-592-8306) with well-defined lines of authority and 

responsibilities. Members of the Hawaii Marine Mammal Stranding Network have 

responded to events such as monk seal entanglements in fishing gear, vessel grounding, 

and the presence of females and pups on public beaches in the main Hawaiian Islands. 

The Hawaiian Islands Response Group V.P. (Marlee Breese, 808-259-5268 or 808-291-

6434) is a key contact in the stranding network of Hawaii. 

 

4.5 Establishing Memoranda of Understanding (MOUs) with Cooperators 

 The U.S. Navy and the U.S. Coast Guard have provided logistical support to the 

Marine Mammal Research Program in past years on a not-to-interfere basis. This support 

has included transportation (of personnel, equipment, and supplies) to and from the 

NWHI and medical evacuation of injured field personnel. MOU with these agencies are 

being explored to establish formal mechanisms for support in the event of a monk seal 

UME. Additionally, an Oil Spill Response Plan is in the final stages of review and will be 

available soon. 

4.6 Pre-Planning for Sample Analyses 

 

 The Marine Mammal Research Program has designated several laboratories 

(Appendix H-3) for analyses of samples collected from Hawaiian monk seals. In addition, 

an epidemiology sampling program provides useful baseline data for comparison in the 

event of a MMUME (Aguirre et al., 1999; Aguirre, 2000). The epidemiology sampling 

program is reviewed periodically (e.g., September 11-12, 2000 Hawaiian Monk Seal 

Health Studies Workshop; November 2001 conference call coordinated by Dr. Robert 

Braun, monk seal health studies coordinator) and changes are implemented as needed (cf. 

Appendix E-1; following the 2000-2001 abortion investigation at Laysan Island, 

sampling and necropsy protocols were modified to improve the efficiency of UME 

investigations). 

5. Recognizing an Unusual Event 

 The following criteria for defining a MMUME are given in the Marine Mammal 

Health and Stranding Response Act (Wilkinson, 1996; Dierauf and Gulland, 2001): 

• It is unexpected; 

• It involves a significant die-off of a marine mammal population; or 

• A small number of a severely endangered marine mammal species appear to be 

affected; 

• Demands an immediate response. 
 11 

 During the 2000 abortion investigation at Laysan Island, the following additional 

criteria were used: increased mortality compared to prior records, mortalities occurring 

in a localized area, and mortalities associated with abnormal behavior patterns (Antonelis 

et al., 2001). 

5.1 Guidelines for a Graduated Response 

 Current Hawaiian monk seal research activities include population monitoring, 

epidemiology program sampling (live and dead seals), documentation of injuries (e.g., 

wounds caused by conspecifics or predators) that might affect an individual seal’s 

survival, monk seal feeding habits studies, and documentation of the use of marine 

habitats by monk seals using satellite-linked telemetry and underwater cameras 

(“CRITTERCAM”). Research conducted as part of the Hawaiian Monk Seal Assessment 

has generated baseline data on expected numbers and timing of births and deaths at monk 

seal colonies; long-term tagging and monitoring programs have generated individual life 

history data for approximately 90% of the population. The Hawaiian Monk Seal Health 

and Disease Program maintains a frozen serum bank and a biomedical database indexed 

by island and by individual seal. NMFS field camp personnel and others (e.g., USFWS 

refuge biologists) are trained to recognize and report live monk seals exhibiting abnormal 

behavior or appearance, as well as any dead seals encountered to the Marine Mammal 

Research Program Leader (Dr. George Antonelis) and the program’s designated 

veterinarian and coordinator of the monk seal health studies (Dr. Robert Braun). Early 

recognition and rapid reporting of a potential MMUME provide time to alert key 

individuals and cooperators (including the NMFS Regional Office, NMFS National 

MMUME Coordinator, and the NMFS Office of Protected Resources), conduct 

preliminary studies and gather additional data, assess capability for response, open lines 

of communication, and identify and refine (as necessary) protocols for response (Geraci 

and Lounsbury, 1997). 

 Baseline data collection and population monitoring of other species occurs seasonally 

(in some cases, year round) in the NWHI and may assist in early recognition and 

reporting of an UME. Fish censuses are conducted annually (and in some cases, monthly) 

in many locations in monk seal habitat as part of ongoing coral reef research programs. 

The U.S. Fish and Wildlife Service conducts surveys at least annually of sea turtle 

nesting activity and abundance and distribution of resident and migratory birds. Yearround camps are maintained at some locations in the NWHI (e.g., Midway Atoll, Laysan 

Island, FFS), staffed by federal and state wildlife biologists. An ongoing photoidentification project is documenting occurrence and behavior of Stenella longirostris at 

Midway Atoll, Kure Atoll, and Pearl and Hermes. The NMFS Southwest Fisheries 

Science Center is surveying cetacean populations along transects near the Hawaiian 

Archipelago. An increase in the number of stranded marine mammals or sea turtles would 

be detected by the NMFS Regional Stranding Coordinator. 
 12 

5.2 Protocol for Initiating Unusual Event Designation 

 

 As specified in Title IV of the Marine Mammal Health and Stranding Act, the NMFS 

Pacific Area Protected Species Program Coordinator/Regional Stranding Coordinator (or 

his or her designee; e.g., the Leader of MMRP) will contact the NMFS National 

MMUME Coordinator if dead or moribund monk seals are observed unexpectedly. This 

contact initiates a chain of events illustrated in a flow chart by Dierauf and Gulland 

(2001, Fig. 1, p. 75), reprinted as Appendix I-1 and briefly summarized here. The NMFS 

National MMUME Coordinator will alert the members of the WGUMMME, who will 

require information on the background levels of morbidity and mortality to decide 

whether or not a MMUME is taking place (data to be obtained from prior records; 

Wilkinson, 1996; p. 18). For example, is the age and sex composition, behavior or body 

condition of the stranded seals different from what occurs normally in that geographic 

location or at that time of year? If the Working Group decides that a MMUME is 

occurring, the MMUME National Coordinator informs the Regional Stranding 

Coordinator, designates an On-Site Coordinator (though the Secretary of Commerce), and 

transfers responsibility for action to the On-Site Coordinator. 

6. Notification of Personnel and Agencies 

 The On-Site Coordinator will notify and mobilize federal, state, and other authorized 

Hawaiian monk seal rescue and recovery program personnel, possible Response Team 

members, and pre-identified laboratories and MOU cooperators promptly. The On-Site 

Coordinator also will notify other federal agencies, state wildlife resource agencies, local 

government agencies, stranding network personnel and the Leader of the Monk Seal 

Recovery Team (see Section 3.2 above). Contact information for agencies and 

individuals to be notified are included in Appendices H-1 through H-4; these lists are 

updated at least annually by MMRP. Additional notifications of other stakeholders will 

be made as needed; for example, the Hawaii Department of Public Health will be notified 

if a serious human health hazard exists on one of the MHI (e.g., if a serious zoonotic 

disease is identified or suspected). 

7. A Tailored Response 

7.1 Objectives and General Guidelines 

 

 The On-Site Coordinator will work with the WGUMMME to determine and initiate a 

course of action to protect the public health and welfare, identify the cause(s) of the event 

(including possible contributing factors such as malnutrition or contaminants), minimize 

deaths and provide for the rehabilitation of individual animals, and determine the impact 

of the MMUME on the population (Wilkinson, 1996; Geraci and Lounsbury, 1997; 

Dierauf and Gulland, 2001). 
 13 

7.2 Responding to an Event of Known Cause 

 

 Wilkinson (1996, Table 1; Appendix I-3) and Geraci and Lounsbury (1997) describe 

the steps to be taken (including continuing consultation with the WGUMMME) in 

response to a MMUME of known cause. Chain of custody procedures will be 

implemented for all data and samples collected in response to a MMUME (Wilkinson 

1996; Appendix G-6), even if the cause is already known or suspected. Pre-identified 

risks to Hawaiian monk seals include biotoxins such as ciguatera, anthropogenic 

chemical spills, abortion “storms,” viral epizootics (e.g., morbillivirus), and humanrelated trauma (e.g., entanglement). 

• Naturally occurring toxins (e.g., harmful algal bloom, ciguatera). The NOAA 

Marine Biotoxins Program has posted a Flow Diagram for Suspected Marine 

Biotoxin Incidents 

(www.chbr.noaa.gov/CoastalResearch/Pictures/AnalyticalResponseTeam/FlowDi

agram.gif); this will be used to guide response to a suspected harmful algal 

bloom. Specimen collection protocols for specific biotoxins are presented in 

Rowles et al. (2001:468, Table 5). See Appendix E-2. 

• Anthropogenic chemicals (e.g., oil or other chemical spill). Procedures will be 

consistent with the contingency plans developed by the U. S. Coast Guard (28 

August 2001 update of the Hawaii Area Contingency Plan,

www.uscg.mil/d14/units/msohono/HACP1/index.htm; Oil Spill Field Operations 

Guide, www.uscg.mil/d14/units/msohono/ics/fog/index.htm). Specimen 

collection protocols for chemical pollutants are presented in Rowles et al. 

(2001:466; Table 4). See Appendix E-3. 

• Abortion. Four aborted fetuses were observed over a 4-week period at Laysan 

Island in 2000. It is not known if these carcasses represented a true increase in 

mortality or simply an increase in survey effort. Baseline data on morbidity and 

mortality are sparse for this time of year because NMFS field camps at Laysan 

Island typically are established several weeks later. Results of the investigation 

into this event were inconclusive. Sixteen seals were sampled, including one adult 

female thought to have aborted a fetus. Laboratory results (including tests for 

Chlamydia sp., Brucella sp., Leptospira sp., morbillivirus and herpesvirus) were 

consistent with baseline serologic test results obtained during previous 

epidemiology studies on Hawaiian monk seals. Five placentas (from surviving 

pups) were examined and considered to be normal. No gross lesions were noted 

during one post-mortem exam on an adult female; histopathology revealed only 

changes consistent with advanced age. However, because abortion-causing 

organisms are a recognized threat to monk seals, genital swabs were added to the 

standard epidemiology and necropsy sampling protocols in 2001 (cf. Appendix E1). 

• Infectious agents (e.g., morbillivirus). Live capture and isolation of affected 

individuals (e.g., while laboratory analyses are pending) may be necessary if an 

epizootic is suspected. Osterhaus et al. (1998) proposed that vaccination be 

considered in the management and conservation of Mediterranean monk seals. 

The Marine Mammal Research Program is reviewing the potential risks and 
 14 

benefits of morbillivirus vaccination in the face of an outbreak in Hawaiian monk 

seals. 

• Human-related trauma (e.g., entanglement). Chain of custody procedures 

(Appendix G-6) will be implemented for all data and samples collected in 

response to an MMUME involving human-related trauma. In addition, NMFS 

Evaluation of Human Interaction forms (Appendix G-5) will be completed for 

affected animals. 

7.3 Investigating an Event of Unknown Cause

 

 Wilkinson (1996, Table 1; Appendix I-3) and Geraci and Lounsbury (1997) describe 

the steps to be taken (including continuing consultation with the WGUMMME) in 

response to a MMUME of unknown cause. Initial data collection will follow existing 

monk seal protocols (e.g., Appendices A through D, G and H) to ensure that information 

collected during the MMUME are comparable to baseline data. As the event progresses 

and at the recommendation of the WGUMMME, additional sampling protocols may be 

implemented (cf. Appendix E). 

 If no cause is readily apparent, the investigation may include analyses of other species 

in the area of the monk seal UME in order to identify or validate possible causes (see 

Section 5.1 for a description of monitoring programs for other species). For example, 

harmful algal blooms often affect a number of plantivorous and piscivorous species (e.g., 

crustaceans, fish, seabirds, sea turtles). The Marine Mammal Research Program is 

preparing cooperative agreements with agencies that oversee these species and resources 

(e.g., USFWS Refuge Managers, NOS-NWHI Coral Reef Reserve, State of Hawaii 

Department of Land and Natural Resources, NOAA National Marine Sanctuary Program 

personnel) to expedite such investigations in the event of a monk seal UME. 

 Under certain conditions (Wilkinson, 1996; p. 41), and if it does not inhibit the 

ongoing investigation, NMFS may accommodate requests from independent researchers 

for tissue samples and data to address other scientific questions that may or may not be 

related to the MMUME. Chain of custody procedures will be implemented for all data 

and samples collected in response to a MMUME (Wilkinson, 1996; Rowles et al., 

2001:452 [Fig. 1]). 

7.4 Determining "The End" 

 

 The WGUMMME, in consultation with the On-Site Coordinator, will determine when 

a MMUME has ended. The end point of a MMUME may be as difficult to pinpoint as the 

onset, particularly if the decline in morbidity or mortality is gradual. If there is a seasonal 

component to the event (e.g., an abortion ‘storm’), investigators may not be convinced 

that the MMUME is over until the following year, when it is possible to assess whether 

or not values (e.g., number of premature pups observed) have returned to historic or 

baseline levels (when compared to prior records, cf. Wilkinson, 1996; p. 18). 
 15 

7.5 Post-Event Assessment and Monitoring 

 Once the end point of a MMUME has been determined, the On-Site Coordinator will 

prepare a report for submission to the WGUMMME containing the results of all analyses 

conducted during the investigation, an assessment of the impact of the MMUME on the 

Hawaiian monk seal population, and recommendations for future monitoring (Wilkinson, 

1996). In some cases, post-event monitoring will return to pre-event levels (i.e., the Monk 

Seal Assessment and Health and Disease programs of MMRP). In other cases, protocols 

may be modified (cf. Appendix E) and additional recovery or population monitoring 

activities may be incorporated into existing research and management programs. 

8. Communication and Reporting 

 The On-Site Coordinator must maintain a regular communication and reporting 

schedule with the Response Team Leader and the WGUMMME. Telephone or email 

briefings every 1 to 2 days will enable the On-Site Coordinator to recognize and address 

personnel, equipment, supply, and data needs of the response team and to ensure that 

critical data and samples are being collected. The Response Team Leader should submit a 

brief written (e-mail) report to the On-Site Coordinator once a week, summarizing the 

status of the investigations and noting unresolved problems. The On-Site Coordinator 

should forward this report with whatever additional calculations, maps, histograms, or 

laboratory analysis results are available to the WGUMMME. The On-Site Coordinator 

and WGUMMME may invite additional experts to review and discuss these reports and 

will determine what additional efforts are needed to respond to and investigate the 

MMUME. 

 The On-Site Coordinator will also provide regular official reports on the progress of 

the investigation to appropriate federal, state, and local agencies and to the Hawaiian 

Monk Seal Recovery Team. The media contact appointed by the On-Site Coordinator 

will keep the NMFS Office of Public Affairs informed, prepare press releases as needed, 

respond to media requests for interviews, and address public concerns and inquiries. 

9. Public Health Concerns 

 Public health and welfare are the first priorities in responding to a MMUME 

(Wilkinson, 1996). The On-Site Coordinator will ensure compliance with safety 

guidelines (e.g., § 300.150 of the National Oil and Hazardous Substances Pollution 

Contingency Plan [40 CFR Part 300] specifies the type of training required for personnel 

responding to an event involving oil discharges or hazardous chemical releases). 

 Only properly trained and equipped personnel will participate in a MMUME response. 

Safety training programs conducted annually for field personnel working with Hawaiian 

monk seals include animal handling skills, first aid, and hygienic precautions such as the 

Monk Seal Clean Protocol and hazardous waste procedures. Additional training (e.g., 

boat handling, aviation safety) will be provided as needed. 
 16 

 Carcasses may be left on the beach to decompose naturally unless a transmissible 

pathogen or serious toxin is suspected. In this case, the carcass may be buried, taken to a 

sanitary landfill, or incinerated. 

 If it is necessary to isolate or quarantine live animals during a MMUME, existing 

protocols developed for Hawaiian monk seals will be implemented (Appendix D). 

10. Resources: Location and Utilization 

10.1 Introduction 

 Although Hawaiian monk seals do occur on the MHI, the majority are found on or 

around the NWHI, a chain of islands and atolls extending hundreds of kilometers. Some 

of these sites are accessible only by boat, and weather and sea conditions often prevent 

landing. Time to mount a response to a monk seal UME is likely to range from a few 

hours (e.g., Kauai) to a week or more (e.g., Laysan Island). 

 The presence of other protected species (e.g., nesting seabirds) may affect the 

activities of UME response personnel. For example, U.S. Fish and Wildlife Service 

quarantine protocols are in effect at some islands requiring special care and treatment of 

clothing, footwear, equipment, and supplies. Access to island interiors may be restricted 

at some islands to prevent disturbance of sensitive animal or plant species. 

10.2 Equipment and Logistics 

All supplies and equipment needed for a limited MMUME response are 

maintained at the Kewalo Research Facility in Honolulu, including capture and 

handling gear (including supplies and equipment for chemical immobilization and 

emergency cardiopulmonary resuscitation), biomedical sampling kits for up to 50 

monk seals, and a portable laboratory. Equipment lists are included in Appendix 

F. 

 Charter flights can be arranged to Midway Atoll, Tern Island, or FFS. Vessel charters 

can also be arranged for logistical support. A list of charter companies is maintained by 

MMRP. The U.S. Coast Guard (Office of Aids to Navigation for ship transport, CGAS 

Barbers Point for air transport) and U.S. Navy have provided emergency transport and 

equipment drops on occasion. MOU with these agencies are being explored to establish 

formal mechanisms for support in the event of a monk seal UME. The NOAA ship Oscar 

Elton Sette and other NOAA research vessels could be diverted from other activities near 

the Hawaiian Archipelago to support a monk seal UME investigation if approved by the 

Director of PIFSC or the Sanctuary Office. 

10.3 Information and Data Resources 

 

 Contact information for key personnel (e.g., potential Response Team members) is 

included in Appendix H. MMRP maintains current contact lists and protocols, and the 
 17 

monk seal assessment program maintains a database of relevant historical information 

such as population counts (beach counts, aerial surveys) and pup mortality rates. Up to 

about 90% of all seals in the NWHI are individually identified each field season. 

Reproductive histories are known for a majority of females at Laysan and for many seals 

at Lisianski and FFS. Age-specific survival is well known for nearly all animals born in 

the past 20 years or so. The Monk Seal Health and Disease Program maintains a baseline 

biomedical database for Hawaiian monk seals, including hematology and serum 

biochemistry reference ranges, seroprevalence of potential pathogens, gross necropsy and 

histopathology reports, and parasite surveys (cf. Aguirre, 2000). Individual biochemical 

serum test results are available for almost 25% of the population. 

10.4 Development of Temporary Animal Housing and Emergency Facilities 

 Rehabilitation facilities for Hawaiian monk seals are maintained at the NMFS 

Kewalo Basin site (pools sufficient to hold 3-5 monk seals), although the ability to isolate 

seals is limited. The Marine Mammal Research Program has construction plans, 

equipment and supply lists, and approximate cost estimates for temporary holding 

facilities that could be erected in remote locations in the NWHI. Gilmartin et al. (in 

review) describe holding pens (35-60 square meters), incorporating sandy haul-out areas 

and shallow seawater access. 

10.5 Funding 

 The MMUME Fund, established in Title IV of the Marine Mammal Protection Act, is 

an interest-bearing account in the Federal Treasury to be used exclusively for costs 

associated with preparing for and responding to MMUMEs (Dierauf and Gulland, 2001). 

In the event of a MMUME, funds are made available through the National Contingency 

Plan (Wilkinson, 1996; Dierauf and Gulland, 2001). Post-UME monitoring is not 

supported by the MMUME Fund, but other federal sources (e.g., Prescott Grant) are 

available for possible funding. 

11. Response and Investigation Protocols 

11.1: Rescue and rehabilitation (Appendix D) 

11.2: Release of rehabilitated monk seals after a MMUME 

 Section 402(b) of the Marine Mammal Protection Act contains guidelines for release 

of rehabilitated animals. The WGUMMME will be consulted to determine whether 

additional event-specific release criteria are needed. The health of the wild population 

will take precedence over the health or fate of an individual monk seal (Wilkinson, 1996). 

11.3: Procedures for Carcasses (Appendix B) 

11.4: Tissue Sample Collection (Appendices A, B, G) 
 18 

11.5: Environmental Assessment and Sampling 

Environmental data for the Hawaiian Islands (e.g., sea surface temperature, wind 

speed, wave height, and primary productivity) are collected by a variety of national and 

international scientific agencies. Wind data used for estimates of oceanic (Ekman) 

transport and ocean mixing (Reynolds) for the period of August 1991 through January 

2001 come from the Advanced Microwave Instruments on ERS-1 and ERS-2 spacecraft. 

These data are provided courtesy of the European Space Agency. The source of wind 

data since January 2000 is the Seawinds sensor on the QuikSCAT spacecraft, provided 

courtesy of the NASA Jet Propulsion Laboratory (an element of the California Institute of 

Technology). The multichannel sea surface temperature (MCSST) data set was developed 

by the University of Miami and is distributed by the NASA Jet Propulsion Laboratory 

PODAAC (JPL PODAAC) administered by the California Institute of Technology. 

Altimetry data also are available from JPL PODAAC. Near real-time data were provided 

by the NOAA/NESDIS Office of Research Applications until 2001; these data are now 

available from the U.S. Navy Fleet Numerical Meteorological and Oceanographic Center 

in Monterey, California. Chlorophyll a data will be obtained through the SeaWIFS and 

MODIS, NASA Active Archive Center at the Goddard Space Flight Center. 

11.6: Packaging and Shipping Samples (Appendices A and H) 

11.7: Keeping Records and Maintaining Chain of Custody (Appendix G-6) 

11.8: Surveys 

 Hawaiian monk seal population assessment field research is conducted at the six main 

breeding subpopulations each year. During this field effort, researchers conduct regular 

surveys of the monk seal’s terrestrial habitat, noting information on individual animal 

identity, location, size class, sex, coarse body condition, and any other notable 

information such as reproductive condition, nursing pair associations, molt status, 

wounding, apparent illness, mortalities, etc. Field research is typically conducted on the 

following schedule: 

Laysan I., Lisianski I., Midway Atoll: mid-March to mid-July 

Kure Atoll, Pearl and Hermes Reef: mid-May to mid-July 

French Frigate Shoals: mid-May to mid-August

 At Laysan and Lisianski Islands, researchers conduct daily surveys of the island 

perimeter. At the remaining multiple-islet atolls, surveys are typically conducted at least 

weekly, weather permitting. 
 19 

12. Acknowledgments 

 We thank the Working Group on Unusual Marine Mammal Mortality Events for 

guidance in preparation of this report. We are particularly grateful to Drs. F.M.D. 

Gulland, A. Hall, O. Nielsen, T.J. Ragen, B.S. Stewart, and J. Whaley for providing 

detailed comments on various drafts of this report. We thank J. Kamiya, J. Kendig, F. 

Fiust, and D. Yamaguchi of PIFSC, NOAA Fisheries for assistance with manuscript 

preparation and editing. 

13. References 

Aguirre, A.A. 2000. Health assessment and disease status studies of the Hawaiian monk 

seal (Monachus schauinslandi). Honolulu Lab., Southwest Fish. Sci. Cent., Natl. Mar. 

Fish. Serv., NOAA, Honolulu, HI 96822-2396. Southwest Fish. Sci. Cent. Admin. Rep. 

H-00-01, 44p. 

__________, J.S. Reif and G.A. Antonelis. 1999. Hawaiian monk seal epidemiology 

plan: Health assessment and disease status studies. U.S. Dep. Commer. NOAA Tech. 

Memo. NOAA-TM-NMFS-SWFSC-280, 63 p. 

Alcorn, D.J. and J.R. Henderson. 1984. Resumption of nursing in “weaned” Hawaiian 

monk seal pups. ‘Elepaio 45:11-12. 

Alcorn, D.J. and A.K.H. Kam. 1986. Fatal shark attack on a Hawaiian monk seal 

(Monachus schauinslandi). Mar. Mamm. Sci. 3:313-315. 

Antonelis, G., B. Ryon, R. Braun, T. Spraker, J. Baker and T. Rowles. 2001. Juvenile 

Hawaiian monk seal (Monachus schauinslandi) unusual mortality event in the 

Northwestern Hawaiian Islands. Pp. 7-8 (abstract) In: Proceedings of the 14th Biennial 

Conference on the Biology of Marine Mammals, Vancouver, Canada. 

Balazs, G.H. and G.C. Whittow. 1979. First record of a tiger shark observed feeding on a 

Hawaiian monk seal. ‘Elepaio 39:107-109. 

Banish, L.D. and W.G. Gilmartin. 1992. Hematology and serum chemistry of the young 

Hawaiian monk seal (Monachus schauinslandi). J. Wildl. Dis. 24:225-230. 

Bernardelli, A., R. Bastida, J. Loureiro, H. Michelis, M.I. Romano, A. Cataldi and E. 

Costa. 1996. Tuberculosis in sea lions and fur seals from the south western Atlantic 

coast. Rev. Sci. Tech. Int. Off. Epizootics 15:985-2005. 

Boness, D.J. 1990. Fostering behavior in Hawaiian monk seals: Is there a reproductive 

cost? Behav. Ecol. Sociobiol. 27:113-122. 
 20 

Caretta, J.V., M.M. Muto, J. Barlow, J. Baker, K.A. Forney, and M. Lowry. 2002. U.S. 

Pacific Marine Mammal Stock Assessments: 2002. U.S. Dep. Commer. NOAA Tech. 

Memo. NOAA-TM-NMFS-SWFSC-3346, 286p. 

Cousins, D.V., S.N. Williams, R. Reuter, D. Forshaw, D. Chadwick, D. Coughran, P. 

Collins and N. Gales. 1993. Tuberculosis in wild seals and characterization of the seal 

bacillus. Aust. Vet. J. 70:92-97. 

Dailey, M.D., R.V. Santangelo and W.G. Gilmartin. 1988. A coprological survey of 

helminth parasites of the Hawaiian monk seal from the Northwestern Hawaiian Islands. 

Mar. Mamm. Sci. 4:125-131. 

DeLong, R.L. 1978. Investigations of Hawaiian monk seal mortality at Laysan, 

Lisianski, French Frigate Shoals, and Necker Island, May 1978. NOAA/NMFS, 

Northwest Alaska Fish. Center, Seattle WA, Interim Rept. (Cited in Kenyon 1981.) 

DeLong, R.L. and W.G. Gilmartin. 1979. Ciguatoxin feeding experiment with a model 

phocid. Monk Seal Workshop Report, U.S. Marine Mammal Commission, 30 Aug. 

1979. Proc. Rept. Nov. 1979. (Cited in Kenyon 1981.) 

Dierauf, L.A. and F.M.D. Gulland. 2001. Marine mammal unusual mortality events. Pp. 

69-81 In: L.A. Dierauf. and F.M.D. Gulland (eds.), CRC Handbook of Marine Mammal 

Medicine, Second Edition. CRC Press, Boca Raton. 

Ewalte, D.R., J.B. Payeur, B.M. Martin, D.R. Cummins and W.G. Miller. 1994. 

Characteristics of a Brucella species from a bottlenose dolphin (Tursiops truncatus). J. 

Vet. Diag. Invest. 6:448-452. 

Fauquier, D., G. Gulland, M. Haulena, L.J. Lowenstine and M. Dailey. 1998. Northern 

fur seal (Callorhinus ursinus) strandings along the central California coast over twentytwo years, 1975-1977. In: Proceedings of the 29th Annual Conference, International 

Association for Aquatic Animal Medicine, San Diego, CA. 

Forshaw, D. and G.R. Phelps. 1991. Tuberculosis in a captive colony of pinnipeds. J. 

Wildl. Dis. 27:288-295. 

Forsyth, M.A., S. Kennedy, S. Wilson, T. Eybatov and T. Barrett. 1998. Canine 

distemper virus in a Caspian seal. Vet. Rec. 138:583-586. 

Geraci, J.R. and V.J. Lounsbury. 1997. DRAFT Contingency Plan for Manatee Die-offs. 

Prepared for Florida Department of Environmental Protection, Florida Marine Research 

Institute, Division of Marine Resources, St. Petersburg, FL, 4 September 1997. 

 

Gerrodette, T.M., M.P. Craig and T.C. Johanos. 1992. Human-assisted fostering of 

Hawaiian monk seal pups. ‘Elepaio 52:43-46. 
 21 

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

Honolulu Lab., Southwest Fish. Sci. Cent., Natl. Mar. Fish. Serv., NOAA, Honolulu, HI 

96822-2896. Southwest Fish. Sci. Cent. Admin. Rep. H-87-17, 30 p. 

 __________, R.L. DeLong, A.W. Smith, L.A. Griner and M.D. Dailey. 1980. An 

investigation into unusual mortality in the Hawaiian monk seal, Monachus schauinslandi. 

Pp. 32-41 In: R.W. Grigg and R.T. Pfund (eds.), Proceedings of the Symposium on the 

Status of Resource Investigation in the Northwestern Hawaiian Islands. University of 

Hawaii, Honolulu. UNIHI-SEAGRANT-MR-80-04. 

__________, T.C. Johanos, D.P. DeMaster and J.R. Henderson. In review. Some life 

history effects following efforts to enhance survival of Hawaiian monk seal pups at Kure 

Atoll. 

Golvan, Y.J. 1959. Acanthocéphales du genre Corynosoma Lühi 1904. Parasites de 

mammifères d’Alaska et de Midway. Annales Parasitologie Humaine et Compare 

34:288-321. 

Harvell, C.D. and 12 others. 1999. Emerging marine diseases – climate links and 

anthropogenic factors. Science 285:1505-1510. 

Henderson, J.R. 1985. A review of Hawaiian monk seal entanglements in marine debris. 

Pp. 326-335 In: R.S. Shomura and H.O. Yoshida (eds.), Proceedings of a Workshop on 

the Fate and Impact of Marine Debris. U.S. Dep. Commer. NOAA Tech. Memo. NOAATM-NMFS-SWFSC-54, 580 p. 

__________. 1990. Recent entanglements of Hawaiian monk seals in marine debris. Pp. 

540-553 In: R.S. Shomura and M.L. Godfrey (eds.). Proceedings of the Second 

International Conference on Marine Debris, 2-7 April 1989, Honolulu, HI. 

NOAA/NMFS Tech. Memo NMFS-SWFSC-154, 1274 p. 

Hiruki, L.M., W.G. Gilmartin, B.L. Becker and I. Stirling. 1993. Wounding in Hawaiian 

monk seals (Monachus schauinslandi). Can. J. Zool. 71:458-468. 

Hiruki, L.M., I. Stirling, W.G. Gilmartin, T.C. Johanos and B.L. Becker. 1993. 

Significance of wounding to female reproductive success in Hawaiian monk seals 

(Monachus schauinslandi) at Laysan Island. Can. J. Zool. 71:469-474. 

Jepson, P.D., S. Brew, A.P. MacMillan, J.R. Baker, J. Barnett, J.K. Kirkwood, T. Kuiken, 

I.R. Robinson and V.R. Simpson. 1994. Antibodies to Brucella in marine mammals 

around the coast of England and Wales. Vet. Rec. 141:513-515. 

Johanos, T.C., B.L. Becker and T.J. Ragen. 1994. Annual reproductive cycle of the 

female Hawaiian monk seal (Monachus schauinslandi). Mar. Mamm. Sci. 10:13-30. 
 22 

Johnson, B.W. and P.A. Johnson. 1980. The Hawaiian monk seal on Laysan Island, 

1978. Draft report to the Marine Mammal Commission, Washington, D.C. 20006. 

Johnson, B.W. and P.A. Johnson. 1981. The Hawaiian monk seal on Laysan Island: 

1978. Final Rept. for the Marine Mammal Commission, Contract No. MM8AC008. 

Report No. MMC-78/15. 

Kenyon, K.W. 1972. Man versus the monk seal. J. Mammal. 53:687-696. 

Kenyon, K.W. 1981. Monk seals Monachus Fleming, 1822. Pp. 195-220 In: S.H. 

Ridgway and R.J. Harrison (eds.), Handbook of Marine Mammals, Volume 2 Seals. 

Academic Press, London. 

Kenyon, K.W. and M.J. Rauzon. 1977. Hawaiian monk seal studies, French Frigate 

Shoals, Leeward Hawaiian Islands National Wildlife Refuge, 15 Feb. to 5 Apr. 1977. 

Proc. Rept. Div. Coop. Res., U.S. Fish and Wildlife Service. (Cited in Kenyon 1981.) 

Kenyon, K.W. and D.W. Rice. 1959. Life history of the Hawaiian monk seal. Pacific 

Science 31:215-252. 

Lavigne, D.M. 1999. The Hawaiian monk seal; Management of an endangered species. 

Pp. 246-266 In: J.R. Twiss, Jr. and R.R. Reeves (eds.). Conservation and management 

of marine mammals. Smithsonian Institution Press, Washington. 

Mamaev, L.V., N.N. Denikina, S.I. Belikov, V.E. Volchkov, I.K. Visser, M. Fleming, C. 

Kai, T.C. Harder, B. Liess and A.D. Osterhaus. 1995. Characterization of 

morbilliviruses isolated from Lake Baikal seals (Phoca sibirica). Vet. Microbiol. 44:2-4, 

251-259. 

Miller, D.L., R.Y. Ewing and G.D. Bossart. 2001. Emerging and resurging diseases. Pp. 

15-30 In: L.A. Dierauf. and F.M.D. Gulland (eds.), CRC Handbook of Marine Mammal 

Medicine, Second Edition. CRC Press, Boca Raton. 

Nitta, E.T. and J.R. Henderson. 1993. A review of interactions between Hawaii’s 

fisheries and protected species. Mar. Fish. Rev. 55:83-92. 

Osterhaus, A. and 14 others. 1998. Monk seal mortality: Virus or toxin? Vaccine 

16:979-981. 

Polovina, J.J., G.T. Mitchum, and G.T. Evans. 1995. Decadal and basin-scale variation in 

mixed layer depth and the impact on biological production in the Central and North 

Pacific, 1960-88. Deep-Sea Res. 42(10): 1701-1716. 

Rausch, R.L. 1969. Diphyllobothriid cestodes from the Hawaiian monk seal, Monachus

schauinslandi Matschie, from Midway Atoll. J. Fish. Res. Bd. Canada 26:947-956. 
 23 

Reddy, M.L., L.A. Dierauf and F.M.D. Gulland. 2001. Marine mammals as sentinels of 

ocean health. Pp. 3-13 In: L.A. Dierauf. and F.M.D. Gulland (eds.), CRC Handbook of 

Marine Mammal Medicine, Second Edition. CRC Press, Boca Raton. 

Reidarson, T.H., J. McBain, C. House, D.P. King, J.L. Stott, A. Krafft, J.K. 

Taubenberger, J. Heyning and T.P. Lipscomb. 1998. Morbillivirus infection in stranded 

common dolphins from the Pacific Ocean. J. Wildl. Dis. 34:771-776. 

Ross, H.M., G. Foster, R.J. Reid, K.L. Jahans and A.P. MacMillan. 1994. Brucella

species infection in sea-mammals. Vet. Record 134:359. 

Rowles, T.K., F.M. Van Dolah and A.A. Hohn. 2001. Gross necropsy and specimen 

collection protocols. Pp. 449-470 In L.A. Dierauf and F.M.D. Gulland (eds.), CRC 

Handbook of Marine Mammal Medicine, Second Edition. CRC Press, Boca Raton. 

Stamper, M.A., F.M.D. Gulland and T. Spraker. 1998. Leptospirosis in rehabilitated 

Pacific harbor seals from California. J. Wildl. Dis. 34:407-410. 

Starfield, A.M., J.D. Roth and K. Ralls. 1995. “Mobbing” in Hawaiian monk seals 

(Monachus schauinslandi): The value of simulation modeling in the absence of 

apparently crucial data. Cons. Biol. 9:166-174. 

Whittow, G.C., G.H. Balazs and G.D. Schmidt. 1980. Parasitic ulceration of the stomach 

in a Hawaiian monk seal (Monachus schauinslandi). ‘Elepaio 39:83-84. 

Wilkinson, D.M. 1996. National Contingency Plan for Response to Unusual Marine 

Mammal Mortality Events. NOAA/NMFS Tech. Memo NMFS-OPR-9. 

Woods, R., D.V. Cousins, R. Kirkwood and D.L. Obendorf. 1995. Tuberculosis in a 

wild Australian fur seal (Arctocephalus pusillus doriferus) from Tasmania. J. Wildl. Dis. 

31: 83-86. 

Yochem, P.K. and B.S. Stewart. 2001. Hair and fur. Pp. 548-549 In: W.F. Perrin, B. 

Wursig and J.G.M. Thewissen (eds.), Encyclopedia of Marine Mammals. Academic 

Press, San Diego, CA. 

14. List of Appendices 

A. HAWAIIAN MONK SEAL SPECIMEN COLLECTION 

B. HAWAIIAN MONK SEAL NECROPSY PROTOCOL 

C. EQUIPMENT AND SUPPLIES FOR A SINGLE NECROPSY 

D. HAWAIIAN MONK SEAL RESCUE AND REHABILITATION GUIDELINES 
 24 

E. ADDITIONAL SAMPLE COLLECTION PROTOCOLS 

1. Additional Samples Requested during the Laysan 2001 

MMUME 

2. Specimen Collection Protocols for Specific Biotoxins 

(Rowles et al., 2001: 468) 

3. Specimen Collection Protocols for Chemical Pollutants 

(Rowles et al., 2001: 466) 

F. EQUIPMENT/SUPPLIES CHECKLISTS 

1. Epidemiology Sampling Supply List 

2. Veterinary Medical Kit 

G. SAMPLE FORMS 

1. Monk Seal Specimen Collection Summary Form 

2. Hawaiian Monk Seal Tagging & Chemical Immobilization 

Form 

3. Hawaiian Monk Seal Clinical Examination Form 

4. Hawaiian Monk Seal Field Hematology Report 

5. NMFS Human Interaction Forms 

6. Marine Forensics Chain of Custody Form (Rowles et al., 

2001. Gross Necropsy and Specimen Collection Protocols. 

Pp. 449-470 in Dierauf, L.A. and F.M.D. Gulland [eds] 

CRC Handbook of Marine Mammal Medicine, CRC Press, 

Boca Raton; Figure 1, p.452). 

H. IMPORTANT CONTACTS 

1. Federal, State and Local authorities 

2. Potential Response Team members 

3. Diagnostic Laboratories 

4. Cooperating Experts 

I. FLOW CHARTS 

1. Flowchart and timing of response to MMUME in the 

United States 

2. Flow Diagram for Suspected Marine Biotoxin Incidents 

(NOAA Marine Biotoxins Program) 

3. Flow chart for response to a marine mammal mortality 

event with known vs. unknown cause (Table 1 in 

Wilkinson, D.M. 1996. National Contingency Plan for 

Response to Unusual Marine Mammal Mortality Events. 

NOAA Tech. Memo. NMFS-OPR-9, 118 p.) 
 25 

J. SAMPLE MOUs FROM CONTINGENCY PLAN FOR MANATEE DIE-OFFS 

1. MOU with participants in UME response 

2. MOU with laboratories/individuals performing 

tests/analyses 

K. HAWAII AREA OIL SPILL CONTINGENCY PLAN 

1. Excerpts from 28 August 2001 Update of the Hawaii Area 

Contingency Plan 

(www.uscg.mil/d14/units/msohono/HACP1/index.htm) 

2. Excerpts from Oil Spill Field Operations Guide 

(www.uscg.mil/d14/units/msohono/ics/fog/index.htm) 
 26 
 27 

APPENDICES 
 28 
 29 

APPENDIX A: 

Hawaiian Monk Seal Specimen Collection Protocol 
 30 
 A-1 

Jan 29 2003/RCB 

HAWAIIAN MONK SEAL SPECIMEN COLLECTION PROTOCOL 

Introduction 

The following protocol describes the collection, processing, preservation, packaging, and shipment 

techniques for specimens collected from Hawaiian monk seals. Strict adherence to this protocol is 

necessary to ensure that each specimen is handled and preserved in an appropriate and standardized 

manner. There is an optimal chance to determine the cause and nature of disease and to assess its 

significance, and the resulting analyses are valid and useful to the management of this endangered species. 

Additional references are provided in Section IV. 

SPECIMEN COLLECTION: POPULATION ASSESSMENT FIELD CAMPS 

Specimens to be collected during population assessment work include: 

Scats 

Spews 

Endoparasites 

Skin tissue plugs 

Molt samples 

Placentas 

Ticks 

Skulls 

Additional specimens to be collected during assessment work include specimens collected from necropsies 

(see Hawaiian Monk Seal Necropsy Protocol). Strict adherence to the necropsy protocol is necessary in 

order to obtain the appropriate specimens from dead monk seals. 

I. Specimen Labeling 

All specimen labels must include the following information: 

Seal ID 

Date collected (YYMMDD format) 

Specimen Type/ Sub-type 

Specimen #/sub #, Island/Atoll 

Include the following if there is room (container larger than cryovial): 

Seal size and sex (if known) 

Preservation method 

Collector initials 

When filling out labels, please write legibly and distinguish between "zero" and the letter "0" by drawing a 

line through the zero. Unless noted otherwise, all specimens should have both an inner and outer label. 

Inner labels 

1. Specimens in jars, glass vials, whirlpaks, and ziplocs should have preprinted waterproof labels 

inside containers in case the outer labels fall off or become illegible. Fill out the labels with pencil 

and be sure to position them inside containers so that labels can be read from the outside

without having to remove the labels to read them. 

2. Specimens in cryovials (blood, serum, plasma, certain blubber samples, etc) and teflon vials 

should not have inner labels. 
 A-2 

Outer labels 

1. Jars and glass vials: use preprinted stick-on specimen labels. After filling out the label, seal with 

clear tape. 

2. Whirlpaks and ziplocks: write directly on bags with a sharpie. 

3. Cryovials: write directly on vial using cryoware markers (provided in necropsy kits). Use the 

following markers for all epidemiology samples: 

black marker=serum 

red marker=plasma LTT 

green marker=plasma GNTT 

blue marker=buffy coats 

4. Teflon vials: use pencil to write on the special labels provided with the vials, then put in the 

vials in whirlpaks with labels both in and out of whirlpaks. 

II. Specimen Preservation 

Refer to material safety data sheets (MSDS) for health effects, first aid, and toxicity of chemicals. 

Common Preservation Methods 

PRESERVATION CODE SPECIMEN FORMULA 

Dry DR scats, molts n/a 

95-100% ethyl alcohol AA tissue plugs, molts n/a 

AFA AF parasitology 50 ml 95% ethyl alcohol (not 

denatured), 5 ml of acetic acid, and 

45 ml of 10% formalin (or 10 ml 

formaldehyde and 35 ml of water 

Alcohol/glycerol solution 

(70 + 5) 

AG parasitology 95 ml of 70% ethyl alcohol (not 

denatured) and 5 ml glycerin 

70% ethyl alcohol AL parasitology 265 ml fresh water to 735 ml of 

95% ethyl alcohol 

Polyvinyl alcohol PV parasitology supplied in pre-filled vials 

10% buffered formalin FM placentas, necropsy 

tissues 

100 ml 37% formaldehyde, 800 ml 

fresh water, and 100 ml seawater 

Liquid nitrogen LN necropsy tissues, 

spew, molts, ticks, 

Note: when placing items in liquid nitrogen dewar, BE SURE items are small enough to be removed from 

dewar! 

NEVER STORE SPECIMENS IN REFRIGERATOR/FREEZER USED FOR STORING FOOD 

FOR HUMAN CONSUMPTION 

III. SPECIMEN COLLECTION METHODS 

A. SCATS 

Scats are collected for prey species determination and parasite studies. Collect a minimum of 100 scats at 

each island or atoll. Always wear disposable latex gloves and avoid direct contact with feces. Select 

fresh scats (not hard or chalky white) and eliminate as much extraneous material as possible (e.g., sand, 

coral, leaf litter, etc.) Emphasis should be on quality, not quantity. In addition, collect a minimum of 10 and 

a maximum of 30 PVA fixed scat sub-samples from fresh scats from known animals. Collection efforts 

should be focused on samples from juvenile animals and collection of repeat samples from the same 

individuals. For collection methods, see parasitology section below. 
 A-3 

1. If endoparasites are observed, collect immediately. Rinse, relax, fix, and store as described in 

the parasitology section below. Be sure parasites are assigned the same specimen number as the 

scat from which it is derived, but give each specimen a unique specimen sub-number. 

2. Collect entire scat (or what's remaining if sub-sample(s) for parasitology are taken) in whirlpaks 

or ziplocs. Be sure to include legible specimen labels both inside (using waterproof labels) and 

outside the bags. Take care to ensure that the inside labels are visible from the outside. 

3. Once any parasitology sub-samples have been collected, add 20% liquid detergent, 80% sea 

water solution to scat sample to begin the degreasing process and to kill some of the pathogens. 

Add just enough of this solution to keep samples moist. Remove as much air as possible from bags 

before storing them in buckets. Be sure the samples are stored double bagged and are kept upright 

in buckets to prevent leaks. 

B. SPEWS 

Spews (vomitus) are collected for prey species determination and parasite studies. They may also be used 

to detect ciguatera (a naturally occurring biotoxin found in dinoflagellates) in tissues of certain prey 

species. Spews are often found near seal haul out tracks and may contain partially digested eels, fish, 

lobster carapaces, octopus, or whole prey items. Note: seabirds also spew squid, flying fish, and other prey 

species, so be sure to closely observe seal tracks in the area the spew is found and collected. 

1. If endoparasites are observed in the spew, collect them immediately and preserve as described in 

the parasitology section below. Be sure parasites are assigned the same specimen number as the 

spew from which it was derived, but give each specimen a unique specimen sub-number. 

2. Next, collect a representative sub-sample of spew that is about 300-400 gm (100 gm is 

approximately hamburger sized) for ciguatera analysis. Once the representative sample is 

collected, split this sub-sample up into ~50 gm units and place in whirlpaks. Assign this spew subsample the same specimen number as the spew from which it was derived, but give only ONE 

unique specimen SUB-number to the representative sample, even though it is being split into 

multiple bags. In addition to recording specimen number and sub-number on inner and outer 

labels, note bag number and total number of bags for one sub-sample on each individual bag (1 of 

5, 2 of 5, etc.) Be sure to also record the total number of bags for one spew sub-sample in the 

"notes" field of the specimen collection summary form. NOTE: the splitting of the sample is 

necessary to store the samples in liquid nitrogen. BE SURE the split samples will fit easily into 

and out of the liquid nitrogen dewar. It can be very difficult, if not impossible, to remove large 

items from the dewars due to their internal design. 

3. Once any parasites sub-samples and the ciguatera sub-sample have been collected, store 

remaining spew DRY in a whirlpak or ziplock, double bagged. Clearly label the outside of the 

bag. Be sure to also include a legible specimen label inside (using waterproof labels) the bag. 

Arrange the inside label so that it is visible from the outside. No preservation is necessary. Be 

sure that the specimen is packed so that it is stored upright in buckets for storage and 

shipment. 

C. PARASITES 

Polyvinyl Alcohol (PVA) fixed fecal specimens: Collect a minimum of 10 and a maximum of 30 PVA 

fixed scat samples from fresh scats from known animals. Collection efforts should be focused on samples 

from juvenile animals and collection of repeat samples from the same individuals. 
 A-4 

1. After collecting a fresh scat sample, take a fecal sub sample using the scoop of plastic lid of 

prefilled vials and/or wooden applicator to collect feces from the center of the scat. Avoid 

collecting sand and other non-fecal debris in sample. 

2. Add sufficient fecal material to PVA vials (prefilled with 5 ml of PVA) to bring the liquid 

volume of the vial to 7 ml. Using scoop and/or applicator sticks, thoroughly mix fecal material 

with fixative breaking up as much of the fecal material as possible. Tightly screw cap on, and mix 

thoroughly by shaking 10+ cycles with rapid twisting motions of the wrist. If you do not have 

prefilled vials, the proper ratio is one part feces to two parts fixative. 

Cestodes (tapeworms) and Trematodes (flukes): Trematodes are flatworms with unsegmented, leafshaped bodies. Cestodes are also flatworms, but differ in that they have ribbon-like bodies that are divided 

into segments called proglottids. These proglottids can be observed with the naked eye in scat samples. To 

preserve both types, remove all grossly visible parasites using wooden applicator sticks and rinse in a 

container with sea water or .85% saline solution. Refrigerate or chill on blue ice for at least 4 hours to allow 

the parasites to relax. After they have become flattened and extended they can be fixed in pre-warmed or 

hot AFA (alcohol-formalin-acetic acid) for at least 30 minutes (fixatives can be prewarmed by setting out 

in sun about 1 hour). If parasites are very long, swirl into fixative using applicator stick. After fixing, rinse 

well with fresh water or 70% ethyl alcohol to eliminate acetic acid that can destroy the specimen. Preserve 

and store the parasites in AG. 

Nematodes (roundworms): Nematodes are round in cross section. To preserve, rinse clean in sea water or 

.85% saline and fix directly in warm or hot AG. 

D. TISSUE PLUGS 

After tagging a seal (see "Tagging/Handling Protocol") , remove tissue plugs from the hole punch 

using Q-tips or clean forceps and place both plugs directly into a vial (>3 ml) that has been prefilled 

with 95% ethyl alcohol. Label the vial with seal ID, etc. Upon returning to camp, fill out any other 

necessary information on the label and add a waterproof label to the vial (be sure it is small enough 

to be easily read from the outside should the external label fall off or become illegible). If necessary, 

top the vial off with additional alcohol so the vial is filled to the top with ethyl alcohol. Seal the vial 

with parafilm. If alcohol is not available, cover the tissue with salt and seal it up. Change the salt the 

following day, when most of the water has been driven out. 

E. MOLTS, VIBRISSAE 

1. Molt samples from known individuals should be collected for DNA analysis in the following order 

of priority: 

a. Tear a sample (2cm x 2cm) and place in a 5 ml cryovial. Store in liquid nitrogen (LN). 

b. Place an additional 2cm x 2cm sample in 95% ethyl alcohol in a 5 ml cryovial. 

c. The remainder of the sample can then be stored in a whirlpak at room temperature. 

2. Vibrissae can be stored in whirlpaks at room temperature. 

F. PLACENTAS 

Caution-there is potential for zoonotic disease transmission from seals to humans (see "Tagging and 

Handling" Protocol). TAKE PRECAUTIONS when handling all samples. Wearing disposable latex 

gloves, collect placentas found with or from an "aborted fetus" or perinatal pup death. Examine and collect 

these placenta according to the Hawaiian Monk Seal Necropsy Protocol. Also collect any fresh placentas 

observed in the following order (if not sure whether it's fresh, collect it): 
 A-5 

1. Collect four 5 cm x 1 cm full thickness strips (extending through to include both the fetal and 

maternal sides) representative of normal and any abnormal portions of the placenta. Put 2 of each 

sample in 2 separate containers and fix in FM. 

2. In addition to preserving sections in FM, freeze four 5 cm x 1 cm full thickness strips 

(extending through to include both fetal and maternal sides) representative of any abnormal portions 

of the placenta. CAUTION: BE SURE samples are cut small enough to be removed from the LN 

dewar. 

G. TICKS 

To collect ticks, look under objects (rocks, boards, buckets) near camp or bird colonies. The ticks like moist 

areas and can be found under the surface layer of the soil. Attempt to collect a minimum of 100 ticks 

(25/cryovial), label, and preserve in liquid nitrogen. If you see ticks on any animal that is being restrained 

(e.g., tagging a weaner), collect all ticks on the animal. Label ticks with seal ID of animal from which the 

ticks are collected. Collect ticks as soon as possible-do not wait until the end of season! 

H. SKULLS 

Collect all skulls found, even if seal ID and history are unknown. Preserve dry (see Hawaiian Monk Seal 

Necropsy Protocol)

I. NECROPSY SPECIMENS-see Hawaiian Monk Seal Necropsy Protocol 

SPECIMEN COLLECTION-EPIDEMIOLOGY STUDIES ONLY 

Specimens to be collected for epidemiology studies include: 

Whole blood (in LTT, SST, GNNT, toxicology vials) 

swabs (nasal, oral, eye, rectal, vaginal, prepucial) 

blubber biopsies (for fatty acid analysis and toxicology) 

fecal samples 

1. BLOOD COLLECTION/PROCESSING: Code 1 ideal; Codes 2, 3, 4, 5 useless 

BLOOD COLLECTION 

Blood will be collected from all seals receiving a dose of diazepam and must be authorized by 

the attending veterinarian. 

The following supplies are needed for blood collection: 

betadine/ 70% ethyl alcohol squeeze bottles 

scrub brush 

1.5" and 18 x 3.5" spinal needles (3 per seal) 

20 or 35 cc syringes (3 per seal) 

extension set with "T" (2 per seal) 

serum separator tubes- 6 minimum 

3.0 mL Lavendar (LTT) - 1 

9.0 mL GNTT-2 

sharpie 

cooler with blue ice pack. 
 A-6 

4' x 4" gauze pads or something to pad blood from contact with ice 

styrofoam test tube holde (to keep SST upright in cooler) 

All persons coming into direct contact with the seal or collecting specimens must wear gloves and other 

protective attire (see Clean protocol for handling seals in “Tagging and Handling” section of HMS 

manual). 

Once the seal is restrained and positioned symmetrically in ventral recumbency (lying on its ventrum (or 

belly) with foreflippers tucked to the seal’s sides), locate the vertebral column (spine) by using the thumb 

and another finger to straddle it; palpate the hips and move forward from the front most aspect of the hips 

approx. 2 vertebrae. Thoroughly clean this sampling site with betadine and scrub brush, then rinse with 

70% ethyl alcohol. If the seal rolls on the sampling site, reclean it. 

Before insertion, remove the needle from the plastic case, and if using a 3.5" needle, remove the stylet and 

plastic piece around hub. Hold needle from hub only (i.e., do not contaminate the needle by touching the 

shaft). Protect the needle with your hand in a position so that the needle does not come into contact with 

anything during restraint. 

Verbally inform restrainers that you are inserting the needle (“sticking”) so they can be prepared in case the 

seal jumps. 

Use the thumb or index finger as a guide to insert the needle between or just to the side of the dorsal 

spinous processes of the lumbar vertebrae (1) and into the bilaterally divided extradural vein (2) that 

overlies the spinal cord (Fig. 1). As the needle is inserted, feel it moving through skin, blubber, and muscle 

until you feel it pop through the membrane of the extradural sinus. You should now see blood rising to the 

hub of the needle. To move the needle once it is inserted, pull it up part way (until it is almost, but not 

completely, out) and then push it back in again. Do not use a slicing motion to move the needle around 

because it may cause tissue and nerve damage. 

Initially, diazapam will be injected. Slow IV administration over a few seconds is preferred over a bolus 

injection. If the animal rolls or otherwise endangers itself with the needle, remove it as soon as possible. If 

the animal does not move significantly and then becomes sedate, leave the needle in place and this needle 

can be used to collect blood from the animal. If the needle is contaminated or removed for any reason, 

insert a new needle before collecting blood. 

If you need to reinsert the needle do it at a different spot than the original location. The angle of the needle 

may vary from a 45 to 90 degree angle forwards or backwards from the dorsal surface of the animal. If the 

needle hits bone you know you are in the right area, so just reposition it. It is best to feel your way through 

the layers and stop before hitting bone, as this can cause the animal pain. 

When the blood vessel is located (you will see blood coming out the top of the needle), attach the extension 

set with syringe to the needle while holding the needle in place with the opposite hand. Make sure to hold 
 A-7 

the needle still and attach the extension set firmly. An assistant should connect the extension set to the 

syringe. 

Collect 35 - 105 cc of whole blood by slowly drawing upward on syringe. Bubbles in syringe indicate your 

needle/extension set or extension set/syringe connections are not secure. This should be corrected as it may 

lead to hemolysis and poor filling. When finished collecting blood, disconnect syringe from the extension 

set, and immediately remove needle from the seal. 

When filling tubes, remove caps (take care to keep inside top of cap from making contact with anything), 

tilt tubes and slowly push blood out of syringe and down side of tube to prevent lysis. First fill LTT and 

GNTT 3/4 full to allow for proper ratio of blood to anti coagulant, immediately replace cap, and mix the 

specimen by gently inverting the tube several times to thoroughly mix additive and blood. Fill scintillation 

or teflon vial (for toxicology) with 5-6 mL of whole blood. Then fill SST with remainder of blood. If 

special tubes for other studies are required, fill them accordingly. 

During restraint, closely monitor respiratory rate, cardiac rate, eyes, and body movements and temperature 

of the seal being handled. Changes in temperature, breathing, and heart rate, unresponsive eyes, seizures, 

muscle weakness or spasms, and rigid/flacid flippers may indicate severe stress or shock. If this happens, 

ABORT the procedure immediately. THE VETERINARIAN ON SITE WILL DIRECT 

EMERGENCY PROCEDURES AS NECESSARY. Emergency procedures may include:

a) moving away from the animal to avoid any added stress 

b) using emergency equipment to establish an airway

c) cooling the seal by drenching with water 

d) loading emergency drugs (see below) 

e) taking careful notes 

f) treating for emergency conditions as necessary, i.e., capture myopathy, dehydration, cardiac arrest, 

vomiting, 

wounds, seizures 

Notify Marine Mammal Research Program Chief of the situation as soon as possible. 

Emergency Drug Doses 

DRUG SEAL SIZE DRAW ADMINISTRATION 

DOPRAM (Doxapram 

hydrochloride; 20 

mg/ml) Pup or juvenile 10 ml 

Give 5 ml, slow IV 

Repeat if needed 

DOPRAM (Doxapram 

hydrochloride; 20 

mg/ml) Subadult or adult 20 ml 

Give 10 ml, slow IV 

Repeat if needed 

ATROPINE sulfate

(1/120 gr) Pup or juvenile 1.5 - 3 ml Give 1.5 ml IV or 3 ml IM 

ATROPINE sulfate

(1/120 gr) Adult 2.5 - 4 ml Give 1.5 ml IV or 3 ml IM 

EPINEPHRINE

(1:1000) All sizes 5 ml 

Give 1 ml/100-200 kg IM or IV, 

repeat prn 

DEXAMETHASONE

4 mg/ml All Sizes Give 10 ml /50-100 kg IM or IV 

FLUMAZENIL

(Romazicon, 0.5 

mg/5/ml) Pup or juvenile 1 bottle (5 ml) 

Give 2.5 ml slow IV (over 60 sec) 

Repeat in 2-3 min if needed 

FLUMAZENIL

(Romazicon, 0.5 

mg/5/ml) Subadult or adult 1 bottle (5 ml) 

Give 2.5 ml slow IV (over 60-90 sec) 

Repeat in 2-3 min if needed 
 A-8 

BLOOD PROCESSING 

LTT, GNTT, and scintillation or teflon vials should be immediately chilled in cooler (avoid contact with 

ice to prevent cell lysis) or refrigerator. Blood in LTT is used for WBC counts. Blood in GNTT is used for 

collection of buffy coats and plasma. The GNTT and LTT (after hematology is completed) should be spun 

in the centrifuge and the plasma pipetted into cryovials. Buffy coats should be collected from GNTT and 

placed in cryovials. If sample collection is done in a remote area, total solids and PCV should be measured 

from plasma collected in LTT. 

The SST tube(s) for collecting serum should sit in an upright position at room temperature or in the shade 

to clot for 15 - 20 minutes, not to exceed 1 hour. Blood should be processed within 4 hours of collection. 

Once the clot in the SST is retracted (i.e., visible separation), centrifuge tubes for 10 minutes at 2500 RPM 

or higher (using the appropriate speed and time according to your centrifuge). The blood will separate into 

3 layers: serum on top (clear, yellowish); middle gel; bottom layer of red blood cells (dark red). Remove 

the serum using a clean pipet and place the serum in a 1.2 mL cryovials, label, and freeze in liquid nitrogen 

or ultrafreeze immediately. The first four 1.2 ml vials of serum (spec sub-#s A-D)will be kept for banking 

so be certain that they are full vials. Avoid using hemolyzed blood for these samples if possible. If any 

vials of serum or plasma that are collected are not full, be sure to record the amount in the vial in the 

comments section of the “Monk Seal Specimen Collection Summary.” 

Dispose of blood collection supplies. Place needles in sharps container and biohazardous waste in 

appropriately labeled bags. 

2. SWAB COLLECTION 

Use sterile dacron polyester fiber tipped swabs with plastic applicators. Swabs may be collected from 

sedated seals in the following order: rectum, nares, medial canthus of eye, lateral commisure of the mouth 

and genital orifice. Collect 2 swabs from each location, place in prelabeled cryovials, break off tips of 

swabs in sterile manner, and preserve in liquid nitrogen. 

If swabs for bacteriology are indicated, they may also be collected at this time. 

3. BLUBBER BIOPSY PROTOCOL: Code 1 ideal; Codes 2, 3, 4, 5 useless 

Supplies needed: 

betadine/alcohol in scrub bottles, scrub brush 

Sterile surgical gloves 

teflon or amber vial for tox biopsy 

 5.0 ml cryovial for Fatty acid biopsy 

disposable scalpel #11 

2 Accupunch biopsy punches 

rat toothed forceps 

scissors 

This technique is to be performed by a veterinarian. Blubber samples will be taken from Hawaiian monk 

seals using a routine procedure requiring sedation (diazapam). Local anesthesia is an option the veterinarian 

may wish to consider, particularly in lightly sedated seals. The blubber biopsy is taken from the lateral 

aspect of the seals pelvic girdle, approximately 5-15 centimeters cranial to the wing of the ileum. The 

incisions should be made to allow proper fluid drainage. Before incisions are made, the area should be 

thoroughly cleaned with *povidone iodine solution and 70% ethanol (take precautions in windy situations 

to avoid getting ethanol in eyes of personnel). Wearing sterile surgical gloves, make two full thickness skin 

incisions using disposable, sterile #11 scalpel blade, through to the level of the blubber, 1-2 cm in length, 

and parallel to the long axis of the animal. introduction of the biopsy punch. Through each incision, insert a 

sterile 6mm biopsy punch. Take a blubber sample through the full depth of the blubber layer (2-5 cm). Use 

sterile or sanitized thumb forceps and/or scissors as necessary, without damaging or contaminating blubber 
 A-9 

tissue. Do not allow gloves to contact tissue! The blubber sample from the first incision should be stored in 

a 5.0 ml cryovial and frozen in liquid nitrogen for fatty acid radioactive isotope analysis. A second blubber 

sample will be collected through the second incision for toxicology analysis. Follow the same technique but 

place samples in glass amber vials and frozen at -20° C as soon as possible. Blubber samples for toxicology 

are to be paired with 5-6mls of whole blood collected during the same restraint period that the blubber 

biopsies are collected. The blood sample should go directly from the withdrawing syringe into scintillation 

vials. The blood should be kept cold and frozen at -20° C as soon as possible. After samples have been 

collected, clean the incisions with *povidone iodine solution. Allow the wound to heal by second intention. 

Suturing is not desirable. When the biopsy is taken in this manner from harbor and grey seals, the incision 

has been found to heal almost completely within a week (S. J. Iverson & W. D. Bowen, pers. Comm., 

1998). Over 200 blubber biopsies have been collected from LEvIS using this method, and no ill effect has 

been reported. 

4. PVA-FIXED FECAL LOOP (PVA-FL) PROTOCOL: Code 1 ideal; Codes 2, 3, 4, 5 useless 

Supplies needed: 

Vials pre-filled with polyvinyl alcohol 

fecal loop 

The fecal loop should be inserted into the anus as far as possible. Using 12-15 ml screw cap vials prefilled 

with 5 ml of PVA fixative and premarked at the 7 ml fill level, add sufficient feces (~2 ml) from the loop to 

bring the liquid volume level to the fill mark. Screw on the cap tightly and mix thoroughly by shaking for 

10+ cycles. Avoid collecting any extraneous materials such as sand, hair, or undigested food items. 

GENERAL GUIDELINES FOR COLLECTION, PRESERVATION, STORAGE, AND HANDLING of 

specimens for diagnostics. Consultation with the laboratory on preferred procedures is required in all 

situations to obtain usable specimens. Tissues and specimens may be also preserved for other diagnostic 

activities, including cytology, immunologic, and nutritional studies (e.g., vitamin A, thiamine, 

radioactive isotopes) as required. 

Diagnostic 

Activity

Type of 

Specimen

Preservation 

Method

Type of 

Container

General 

Observations

Histopathology All tissues and 

lesions 

10% buffered 

formalin 

Leak-proof glass 

or plastic 

Formalin ratio 

10:1; room 

temperature 

Toxicology Organs, blood, 

blubber, ingesta 

Refrigeration or 

freezing 

Sterile glass or 

plastic, metal foil, 

teflon 

Procedures 

change; need good 

records 

Microbiology -

virus, bacteria, 

fungi, rickettsia 

Organs, tissues, 

swabs, lesions, 

body fluids, buffy 

coat 

Refrigeration, 

freezing or 

ultrafreezing 

Sterile cryovials, 

plastic or glass, 

special vials, and 

transport media 

Avoid 

contamination; 

sampling varies 

with disease 

Parasitology Endoparasites* 

Ectoparasites 

Hemoparasites 

AFA or AG 

solution 

AG solution or 

10% buffered 

formalin 

Smears or whole 

Glass or plastic 

Glass or plastic 

Glass slides or 

EDTA tubes 

Room temperature 

Room temperature 

Room temperature 

Refrigerated 
 A-10 

Blood 

Hematology and 

Plasma Analysis 

Whole blood in 

lavender, green, 

and gray top 

vacutainers 

Refrigeration until 

spinning, Liq. N2

after pipetting 

Pipetting of 

plasma into 

cryovials 

Short storage, 

immediate 

processing 

Serology Blood Refrigeration/ 

freeze (-20C) or 

ultrafreeze serum 

Sterile cryovials Handle gently to 

avoid red blood 

cell rupture 

Genetics Skin punch; 

buffy coat and red 

blood cells 

Ethyl alcohol 

or salt; freezing 

or ultrafreezing 

Vials; cryovials, Long-term storage 

PACKAGING AND SHIPMENT 

IMPORTANT: Include original copy of completed Necropsy Report Form when sending necropsy 

specimens back to Honolulu. When sending ANY specimens, include a disk with the latest back-up of 

the specimen data base. 

Notify Honolulu several days in advance before sending specimens back so that arrangements can be 

made to pick them up. 

Update the transport information in the “Monk Seal Specimen Collection Summary” prior to sending any 

samples back to Honolulu (so that the data includes the shipping information). 

All buckets and lids used for storing/transporting specimens should be labeled “SPECIMENS, NO FOOD” 

using a permanent marker. 

Bucket lids should be labeled with specimen types (e.g., “SCATS,” “NECROPSY TISSUES,” “TISSUE 

PLUGS”) 

1. Pack all necropsy tissues and placentas together. Tissue plugs, molts, and PVA vials may be packed 

together. Scats and spews may be packed together, but do not pack any other specimen types with them. 

Be sure all specimens are double bagged, placed upright to avoid leaking, and be sure there is enough 

absorbent material packed around containers in case of leakage. 

2. Be sure to secure nitrogen dewar lid with zip ties or duct tape. Ensure that dewar is secured during 

transport so it cannot tip over. 

3. If sending specimens in a cooler, all individually packaged specimens may be placed together in one 

outer bag and then surrounded by packing materials and the proper coolant (i.e., blue ice, dry ice, etc.). 

Line the shipment container (e.g., bucket or cooler for frozen items) with a plastic bag and place absorbent 

packing material such as newspaper, paper towels, or gauze on the bottom and in between layers of sample 

containers; also fill any remaining space with packing material to secure samples. When shipping frozen 

items via plane or ship, put an ice pack in the cooler; if via ship freeze immediately once aboard the vessel. 

4. Make sure lids on sample containers are secure (use Parafilm or tape if necessary), bags are well sealed, 

and containers are in an upright position. 
 A-11 

5. Tissues that have been properly fixed may be transported without FM. For those specimens which are 

too large to be transported in secondary containers (e.g., GIT specimens in 5 gallon buckets), FM MUST

be drained before transport. The used FM can be drained into carboys which should be clearly labeled with 

both the specimen number and bucket number from which it was derived. Field camp leaders are 

responsible for ensuring that the specimens are refilled with the used FM upon return to Honolulu. 

All buckets containing specimens should be clearly labeled with the following: 

Keep Upright 

Sample type (i.e., scats, spews, necropsy tissue) 

Preservation Method (e.g. 10% buffered formalin, polyvinyl alcohol) 

Island/Year 

Appropriate HazMat stickers 

Caution - hazardous/infectious material (if applicable) 

IV. REFERENCES 

Dierauf, L.A. 1994. Pinniped forensic, necropsy, and tissue collection guide. U.S. Dept. of Commer., 

NOAA Tech Memo. NMFS-OPR-94-3, 80p. 

Geraci, J.R., and V. J. Lounsbury. 1993. Marine Mammals Ashore: A field guide for strandings. Texas 

A&M University Sea Grant College Program, Galveston, Texas, 305 p. 

Winchell, J.M. 1990. Field manual for phocid necropsies (specifically Monachus schauinslandi). U.S. 

Dept. of Commer., NOAA Tech. Memo. NMFS-SWFC-146, 55 p. 
 A-12 

MONK SEAL SPECIMEN COLLECTION SUMMARY INSTRUCTIONS 

For each specimen collected, log information in the “Monk Seal Specimen Collection Summary” found in 

Master Field Log. Enter data into the SPECIMEN database. 

SPECIMEN 

NUM Consecutive 4-digit numbers (0001 - 9999) of samples as they are collected, regardless of 

sample type. These numbers are unique for a given atoll/year. Each sample container 

should have its own specimen number and label. 

SUB-NUM Complete only if a specimen is split. If the specimen is split, assign consecutive 

sub-numbers (aa - zz). A specimen can be split in the field or back in the lab. If 

you have a single specimen 0043, and you split it into two parts, then make one 

part 0043a and the other part 0043b. If 0043a is later split into two parts, make 

one part 0043aa and the other 0043ab. If 0043a has been split so that you have 

0043aa, 0043ab, and 0043ac, and you want to split 0043aa, then leave one of the 

new split parts as 0043aa and make the other part the next available sub-number 

(i.e., 0043ad). Each container must be labeled, and must have a unique 

number/sub-number. 

TYPE Type of specimen collected. 

BB = blubber biopsy, collected from live animal 

 BC = blood cells (remainder in tube after removing plasma or serum) 

 BL = blood (as for virology scint. vials) 

 BU = buffy coat 

 BS = blood smear 

 ES = eye swab 

 FL = fecal loop 

 GI = gastrointestinal tract section taken anywhere from pharynx to anus 

MO = molt 

 NS = nasal swab 

 NT = necropsy tissue (tissues from all organs, includes necropsy blubber 

sample) 

 PC = placenta 

 PL = the 'pluck': heart, trachea, lungs and associated glands 

 PS = plasma 

 RS = rectal swab 

 RT = whole reproductive tract of female or male 

 SC = scats 

 SE = serum (collected in serum separator tube for serology) 

 SK = skeletal (bones or teeth) 

 SP = spew 

 ST = stomach contents 

 TP = tissue plug 

 VS = vaginal swab 

 WB = whole blood 

 WC = whole carcass 

 WH = whole heart 

 ON = other necropsy (non-tissue samples, e.g., bile, urine, aqueous humor) 

 OS = other swab 

 OT = other (indicate type in notes section on back of form) 

NO = none 
 A-13 

SUBTYPE Subtype of specimen collected (i.e., parasite from a spew). Distinguish necropsy 

tissue 

 subtypes (*) for frozen toxicological samples. 

 AD = adrenal (*) 

 BC = blubber chunk (necropsy tissue) 

 BM = bacteriology medium 

 BP = blubber biopsy plug (necropsy tissue) 

 BR = brain (*) 

 BT = blue topped tube (BTT) 

 DN = DNA analysis 

 GI = sample from gastrointestinal tract (*) 

 GN = green topped tube (GNTT) 

 GT = grey topped tube (GTT) 

 KI = kidney (*) 

 LI = liver (*) 

 LT = lavender topped tube (LTT) 

 LU = lung (*) 

 NP = not processed/intact (scats and spews) 

 OL = otolith 

 PA = parasites 

 RT = red topped tube (RTT) 

 SP = spleen (*) 

 SS = serum separator tube (red/grey topped; SST) 

 VM = virology medium 

SEAL ID Record the 4-digit permanent ID number of seal if known. 

TEMP ID Record the temporary ID number (or bleach number) of seal if known. 

SIZE 

P0 = Fetus (aborted, clearly pre-term pup) 

P = Nursing pup 

 P1 = Nursing pup, wrinkles 

 P2 = Nursing pup, no wrinkles 

 P3 = Nursing pup, blimp, black 

 P4 = Nursing pup, molting 

 PS = Nursing pup, molted 

PW = Prematurely weaned/undersized weaned pup (weaned ≤ 2 wks ago and < 90cm girth) 

W =Weaned pup 

I = Immature 

 J = Juvenile 

 J1 = Juvenile I 

 J2 = Juvenile II 

S = Subadult 

S3 = Subadult III 

 S4 = Subadult IV 

A = Adult 

U = seal of unknown size 

SEX M = Male, F = Female, U = Unknown 

SURVIVAL FACTOR NUM Survival factor number. 
 A-14 

COLLECT DATE Date sample is collected (in MMDD format). 

ISLET Two-digit numerical code for the islet from which the sample is collected. 

PRESERVATION METHOD at COLLECTION, during TRANSPORT, and on ARRIVAL in 

Honolulu 

AA = absolute ethyl alcohol (98-100%) 

AF = AFA: 50m1 of 95% ethyl alcohol (not denatured), 5ml of acetic acid, and 

45m1 of 10% formalin (or 10 ml formaldehyde and 35m1 of water) 

 AG = 95mm of 70% ethyl alcohol (not denatured) and 5ml glycerin 

 AL = 70% ethyl alcohol 

 CB = chloroform, BHT mix for blubber biopsies 

 DM = DMSO 

 DR = dry 

 FR = frozen 

 FM = 10% buffered formalin (neutral buffered formalin=NBF) 

 GA = 20% glycerin and 80% ethyl alcohol 

 LN = liquid nitrogen 

 PV = polyvinyl alcohol 

RE = refrigerate 

 RT = room temperature 

 UF = ultrafreeze 

OT = other (indicate method in notes section on back of form, use for whole scat 

 w/detergent) 

 NO = none (sample was discarded and not preserved) 

TRANSPORT MEANS Method of transportation used to ship sample to Honolulu. 

 CV = charter vessel (ship) 

 CP = charter plane 

 MP = military plane 

 OS = Oscar Sette 

 OT = other (indicate method in notes section) 

DATE SENT Date sample sent from the field. 

PROJECT: Code what project the specimens were collected for. 

 UM = UME 

 EP = epidemiology 

 PA = population assessment 

 PS = pup survival 

 ST = satellite tagging 

 CC = critter cam 

 OT = other 

NOTES Blank = no handwritten notes for a sample. 

 1 = handwritten notes for a sample are on the back of form, labeled by sample number. 

Enter 

 these into the Notes field of the database (do not exceed 250 characters). 

 2 = sample collected, analyzed, and discarded. 
 A-15 

INIT Three initials of collector. If no middle initial, use the first and last block. 

The following fields will be filled out later in Honolulu: 

DATE ARRIVED Date sample arrived in Honolulu. 

DIAGNOSTICS 

LAB Location sample sent for analysis. Code as follows: 

 AF = Armed Forces Institute of Pathology (AFIP) 

 CA = University of California-Davis 

 CO = Colorado State University-Fort Collins 

 CV = CVD/IDDEXX 

 GL = Gwen Lowe 

 MK = Michael Kliks 

 NW = NWHRC-Honolulu Station 

 NY = Cornell University-Ithaca, New York 

 OA = Oklahoma Animal Disease Diagnostic Laboratory 

 OS = Oregon State University 

 SC = NMFS- Charleston, South Carolina 

 UH = University of Hawaii 

 US = USDA 

 WA = NMFS- Seattle, Washington 

 WD = Washington Animal Disease Diagnostic Lab 

 YL = Ron Lu (UH) 

 OT = other 

DATE SENT Date sample sent for evaluation. 

EVALUATION Code for the diagnostic tests performed. 

BA = Bacteriology 

BP = Blood profile 

CP = Chemistry profile 

CY = Cytology 

EM = Electron microscopy 

EP = Endocrinology profile 

GE = Genetics 

HP = Histopathology 

IM = Immunology 

MY = Mycology 

NP = Nutritional profile 

PA = Parasitology 

PR = Prey analysis 

SE = Serology 

SM = Blood smears 

TX = Toxicology 

UA = Urine analysis 

VI = Virology 

NE = Necropsy 

OT = Other 

RESULTS Results of diagnostic tests. Codes not yet available. 

DATE TESTED Date of laboratory procedure. 
 A-16 

SPECIMEN STORAGE: 

FACILITY 

 HL= Honolulu Lab (Dole Street) 

KR= Kewalo Research Facility 

LOCATION 

 A2= Annex 2

DH= Diamond Head Lab 

RE= Regular Freezer 

UF=Ultra freezer 

SHELF 

BIN/BOX 

CELL 
 

APPENDIX B: 

Hawaiian Monk Seal Necropsy Protocol 
 
 B-1 

28 Jan 2003/RCB 

HAWAIIAN MONK SEAL NECROPSY PROTOCOL 

NOTE: Read the "Field Manual for Phocid Necropsies (FMPN) (specifically Monachus 

schauinslandi)" and this Necropsy Protocol carefully and several times before performing necropsies. 

Necropsy or post-mortem examination of monk seals is a basic tool for the investigation of disease and 

monitoring the health of wild populations. It can be defined as the systematic examination of the whole 

body, organs, and tissues. Whenever possible, necropsies should be performed by a trained veterinary 

pathologist experienced in recognizing and interpreting lesions and abnormalities. 

General Considerations 

A necropsy has the most scientific value when it is carefully documented. The use of this Necropsy 

Protocol will assist in the documentation and standardization of information which may be valuable in 

determining morbidity and mortality factors within individual seals as well as the population. 

Preventing Disease Transmission 

Avoid direct contact with dead seals to prevent transmission of infectious diseases that are pathogenic to 

humans. Persons performing the necropsy must: 

 1. Wear protective gear, including latex or plastic (untorn) gloves, mask, coveralls/apron and 

 rubber boots. 

 2. Cover your surface wounds with protective dressings before putting on gloves. 

 3. Collect frozen specimens only at locations where freezing is possible. DO NOT STORE 

 SPECIMENS IN FREEZERS/REFRIGERATORS USED FOR HUMAN FOOD.

 4. When the necropsy is completed, wash skin thoroughly with antibacterial soap and wash/sanitize 

 or dispose of all contaminated clothing in a container clearly marked "Biohazardous waste." 

 5. If possible, pull the carcass to high ground (so it won't wash into the water) and bury to avoid 

 attracting predators and disease transmission. 

 6. Scalpel blades must be disposed of in sharps containers. Disinfect instruments with a 50% 

 chlorine bleach solution and thoroughly rinse with fresh water; or wash with soap and fresh 

 water and then rinse with 70% ethyl alcohol. Keep all necropsy supplies clean and dry. 

 7. Seek medical attention for bites, cuts and other injuries and notify physician of their source. 

General considerations in performing a necropsy include: 

1. A complete and careful record must be kept of what is observed during the necropsy. 

 2. First obtain morphometric data, then external samples. 

 3. The order of the Necropsy Report Form follows the sequence of general dissection and 

 examination. See sample collection and preservation sheet. 

 4. Tissues and organs must be examined in a systematic manner. The precise method used for 

 a necropsy is less important than establishing a routine in which each body system is 

 examined fully. 

 5. Once the carcass is opened, take tissue specimens for virology, bacteriology, and 

toxicology (only if a freezer and/or liquid nitrogen is available), then sample for 

histopathology and parasitology. 

 6. The objective is to recognize abnormalities; this requires the ability to distinguish between 

 normal and abnormal tissues. 

 7. Samples of normal and abnormal tissues should be collected for laboratory analyses. 

 8. With an "aborted fetus," perinatal death, or newborn, perform necropsy according to 

 “Fetus” section of necropsy form. 
 B-2 

The ability to obtain reliable data from necropsies depends on the following: 

1. Condition and location of the specimens. 

2. Adherence to detailed protocols. 

3. Number of seals to be necropsied throughout a field season. 

4. Amount of YOUR time available to perform a thorough necropsy. 

5. Care in sample preservation and labeling of specimens. 

6. Care in shipping and storing specimens. 

Decomposed carcasses are unsuitable for histopathology but are useful for describing gross lesions. Brain, 

intestines, liver and other enzyme-rich organs are the first to deteriorate. To date, the etiology of the eye 

condition of the captive seals remains unknown, therefore you should collect eyes first. Also, if the carcass 

is a code 1 (see page 8 for carcass condition codes) and the necropsy is being performed less than 1 hour 

after death has occurred, the brain should be collected second. Take samples from all organs listed, even 

those that appear normal. Tissue specimens must be sufficiently thin (less than 1 cm thick) to allow fixation 

with formalin except for brain and lung. Make parallel cuts ( 1⁄2 cm in thickness) in formalin fixed samples 

to allow preservation. Samples preserved in formalin should be fixed in 10% buffered formalin with 10 

units volume of formalin to 1 unit tissue volume. Throughout the necropsy, refer to the FMPN. 

Specimens for toxicology from live animals may be limited to blood and biopsies of skin and blubber (see 

Blubber Biopsy Technique). When sampling for toxicology, it is important to use standardized sampling 

procedures so that even when low levels of contaminants are present, differences may be attributed to 

biological processes and contaminant exposure and not to variation in the collection process. Tissues from 

dead monk seals must be collected less than 24 hours from time of death. As soon as possible, freeze at 

lowest temperature available. 

 1. Toxicological analyses may be performed for heavy metals, organochlorides, selenium, and dioxin. 

Tissue samples will be taken in duplicate (2 of each, except when noted). 

 2. See FMPN for collection techniques. Use a new, sterile blade for each organ collected. Any stainless 

steel instruments used in contact with tissues should be cleaned with distilled water and rinsed with 

isopropyl alcohol before using if possible. Each specimen should be rinsed with distilled water (if 

possible), wrapped in aluminum foil (with dull side touching specimen), placed in whirlpaks or 

ziplocks, then frozen. 

 3. Collect the following samples by special request only: Biotoxin (ciguatera poisoning) and/or 

 hydrocarbons (oil spills) specimens may be collected at selected sites. The specimens, also collected 

 in duplicate, include: 

a. Blubber: One, 100 gram sample, rinse with distilled water, wrap in Teflon sheet, place in 

whirlpak or ziplock. 

b. Liver: One, 100 gram samples, rinse with distilled water, wrap in Teflon sheet, place in 

whirlpak or ziplock. 

4. Because analyses may involve detecting extremely small amounts of contaminants, care must be 

taken in the collection and processing of these tissues. Avoid salt water, tobacco smoke, bug 

sprays, and other aerosolized foreign materials during collection. 

5. Tissues should be collected as rapidly as possible after opening body cavity to prevent 

 contamination and deterioration. 
 B-3 

TISSUES FOR MICROBIOLOGY: Code 1 ideal; Codes 2, 3 limited; Codes 4, 5 useless 

Collect the following by special request only: For special studies, you may be required to collect 

additional specimens from monk seals. 

Specimen collection for bacteriology and virology is determined primarily by the nature of gross pathologic 

lesions. Samples should be taken aseptically, from external surfaces, body cavities,and internal organs as 

soon as they are exposed. Place swabs in respective transport media and refrigerate at 4° C or place on blue 

ice immediately and freeze upon arrival to laboratory or field camp. If cryovials are available, ultrafreeze 

the swabs with tissue samples in liquid nitrogen. Samples for microbiology are worth the time and effort 

only when tissues are in suitable condition. With an "aborted fetus," perinatal death, or newborn, collect 

specimens according to "Fetus" section of Necropsy Form for freezing and later microbial analysis. 

INSTRUCTIONS 

A. Before conducting a field necropsy, become familiar with the "Field Manual for Phocid Necropsies 

(specifically Monachus schauinslandi)" for explanations of external and internal examination. Refer to 

this manual during the necropsy. Also become familiar with the protocol BEFORE performing necropsies. 

B. Read the "Specimen Collection Protocol" for instructions on collection, preservation, packaging, and 

shipment techniques of samples collected 

C. Use the Hawaiian Monk Seal Necropsy Report Form for all seals found dead during the field season, 

whether or not a necropsy was performed. Record "N/A" for any sections that are not applicable, and state 

what organs/tissues were not examined. At a minimum, identify the appropriate descriptors for each organ 

examined and sample all organs. 

D. If the seal has flipper tags, note the condition of the tags on the census and tag condition forms. Recover 

flipper tags, place in a whirlpak bag and label the bag with animal ID, island, date, and survival factor 

number. Also check for PIT tags with the PIT tag reader. Even if PIT tags were not found during a scan, 

please indicate that a scan was completed. Also note where on the body the scan was performed. Record 

this information on the census sheet as well. If the ID of the seal is unknown or questionable, document any 

specific external lesions, abnormalities, or scar patterns. For seals that appear to have been mobbed, 

examine, describe, and photograph the mobbing injuries, anogenital area, scars, and other distinguishing 

characteristics such as injuries and external lesions. Record all photographs on census form and on the 

Necropsy Report Form as requested. 

TAKE INTERNAL PHOTOGRAPHS ONLY WHEN UNUSUAL CONDITIONS ARE 

NOTED OR IF YOU ARE UNSURE IF IT IS UNUSUAL. If unusual conditions are noted, 

include a size reference (e.g., ruler) and label with seal ID, survival factor number, date, and 

island. Take two photographs, one with the organ in situ (in its anatomical position/location) in 

the body and one with the organ removed from the body and placed on a solid white or light blue 

surface. 

E. Measure axillary girth, dorsal length, and ventral length. Refer to the Necropsy Report Form for seal 

carcass measurements requested. Record measurements on both the TAGGING/HANDLING CARD 

and the Necropsy Report Form. 
 B-4 

F. Carcass evaluation. A carcass condition code system has been developed based on the state of 

decomposition of a monk seal carcass (see page 8). Carcass condition is influenced by many factors 

including disease, body temperature, and environmental temperature. Rigor mortis (stiffening of the body 

following death) may serve as an indicator of carcass evaluation. It can occur within hours in warm 

weather, but is extremely variable. Rigor mortis indicates that a carcass may be in good condition (see 

Code 2 in Necropsy Report Form). 

G. Assume more is better when describing and recording information. The rule here is if in doubt, write it 

down. If unsure whether something is abnormal, state this and succinctly describe. Descriptions should be 

clear, concise, and without personal interpretation. Appropriate tissue preservation along with YOUR 

precise description of findings may allow the identification of causes of death in the population. 

H. Identify the appropriate descriptors for each organ examined. The descriptions provided herein are NOT 

an exhaustive list of terms, but rather a list for your reference. Describe surface, consistency, color, and cut 

surface of both normal tissues and abnormalities or lesions. 

DESCRIPTORS of ORGANS AND LESIONS

Surface: Smooth, rough, shiny, dull, thickened, wrinkled. 

 Consistency: Firm, soft, flabby, dry, wet, fluid-filled, sharp-edged, friable (easily pulverized 

 or crumbled). 

 Color: Translucent, opaque, green, yellow, brown, pink, red, nutmeg (normal pattern of 

 liver), etc. Also comment how color is spread through tissue, if it is 

homogenous or speckled (i.e., pink with specks of red). 

Cut surface: Slice organ several times appropriately and spread apart to look at internal 

 surface. Be sure to describe color of the cut surface. Descriptors include 

swollen, bulging, shiny, dull, eroded, glistening, scaly, pitted, oozing. 

Size: Record in metric system (mm, cm), measure length, width and depth, or 

 diameter of the lesion. Enlarged (hypertrophied), small (atrophied), normal size. 

 Shape: Square, rectangular, triangular, oval, round, cuboidal, spherical, discoid, 

rhomboid, tear- shaped, wedge-shaped, spindle-shaped, irregular, long, slender, 

indented, narrow, lace-like, tortuous, branching, speckled (miliary), flat, raised, 

depressed, shrunken, papillary, cauliflower-like. 

 Distribution: Single discrete lesion (focal), multiple lesions in one location (multifocal), or 

 multiple lesions scattered diffusely throughout the organ or body cavity 

 (diffuse); locally extensive, random, even. 

 Location: Surface, capsule, wall, dorsal or ventral, caudal or cranial, anterior or posterior, 

 medial or lateral, proximal or distal, internal or external, full or partial thickness 

 of a wall of an organ. 

 Color: Additional descriptors may include bright, pale, streaked, blotchy, dark, 

Blanched, mottled yellow with brown specks, brick red, Christmas red, yellow 

with white streaks yellowish white, blotchy red brown, creamy, off-white, 

transparent, opaque. Use simple colors, do not get complicated. 

 Fluid: Clear, cloudy, turbid, thick, thin, bloody, muciod, exudate, dark, tarry. 

 Consistency: Spongy, granular, gel-like, firm, soft, hard, rock-hard, dense, creamy, buttery, 

 brittle lumpy, velvety, warty, tenacious, gritty. 

 Texture on cut 

 surface: Bulging, engorged, granular, nodular, pitted, oozing 

 Odor: None, sweet, sour, rancid, ammonia-like, putrid, fruity, 

 petroleum-like 

I. COLLECT ALL LESIONS. Describe and sample areas that 

appear to stand out in marked contrast to the main body of tissue. Include 

a description of the margins between the normal and abnormal tissue 

Norma

tissue

Lesion

Cut
 B-5 

i.e., sharp line versus vague and gradual, circumscribed, encapsulated. 

When collecting abnormal tissues for histopathology, be sure to include the margin between 

the abnormal and adjacent normal tissue and accurately describe abnormalities (including 

precise location). 

J. Review the completed Necropsy Report Form and file in the 

 Master Field Log. Review census form for sighting of animal as 

 dead, and include survival factor number, photos taken, tag 

 condition, and tags recovered. Make sure that the tagging/handling 

 card, scar card, and tag condition drawing form are complete. 

K. Record specimens collected on the Specimen Collection Summary and assign specimen numbers 

as outlined in the Specimen Collection Protocol. The 2 jars of tissue sets for histopathology 

(HP) should have the same specimen number but a different sub-number. Make sure to check 

the boxes next to the appropriate specimens as collected on the necropsy report form. 

L. Necropsy Report Forms, scar cards, tagging/handling cards, survival factor forms, and photos 

should be returned to the NMFS Honolulu Laboratory. 

M. Clean necropsy tools (you may also need to spray them with WD-40 or LPS) and restock necropsy 

kit so that it is ready for the next necropsy. 

EQUIPMENT AND SUPPLIES FOR A SINGLE NECROPSY 

1. Clipboard, metal 

2. Necropsy Table (Checklist) 

3. Necropsy Report Form(s) 

4. Necropsy Manual 

5. Scarcard 

6. Census Form 

7. Tag/handling card 

8. Survival factor form 

9. Self-sticking pre-printed specimen labels 

(3 sheets) 

10. Waterproof labels ("rite-in-rain" paper, 

pre-cut) (3 sheets) 

11. Pencils (2) 

12. Ball-point pens (optional) (2) 

13. Permanent markers (sharpies) (2) 

14. Ruler (metric) 

15. Metal tape measure (metric) 

16. Tape measure, flexible (metric) 

17. Surgical scissors (2: straight and curved) 

18. Rubber gloves 

chemical resistant - 2 pr. latex 

heavy duty - 6 pr. latex exam 

gloves - 15 pr 

19. Scalpel handles (4: 2 #8, 2 #6) 

20. Scalpel blades (30) 

21. Blubber biopsy punch, 6 mm (2) 

22. Forceps (2 pr.) 

23. Hemostats (1) 

24. Dacron Swabs (5) 

25. Microscope slides 

26. Slide holder for microscope slides 

27. Small vials (—2 x 1⁄2") - 6 ea. 

28. Med. vials (—3' x 2) - 6 ca. 

29. Whirlpak bags (20 ea.) 

30. Ziploc bags (large) 

31. Paper towels 

32. Antibacterial soap/Hand sanitizer 

33. Sharps container 

34. Needles, 1 1⁄2', 18 g (5) 

35. Syringes, 20 cc or 35 cc (5) 

36. Blood tubes, PT, TT (4) 

37. Biohazard bag (2) 

38. Camera with colored slide film 

and/or Digital camera 

39. Shovels 

40. Jars and buckets partially filled 

with 10% NB Formalin 

a. 2 large jars for duplicate 

sets of histopathology 

tissue samples 

b. 1 bucket for entire female 

reproductive tract 

c. 1 bucket for esophagus 

d. 1 5gal. bucket for entire 

gastrointestinal tract from 

beginning of stomach to 

anus 

e. 2 jars for eyes 

f. 2 jars for brain 

41. Hacksaw 
 B-6 

42. Axe 

43. Cryovials (20) 

44. String to tie off beginning of stomach, etc. 

(1 roll) 

45. Cutting board (not on spc. coil. list) 

46. Disposable aprons (3) 

47. Bone shears 

48. Archival tag labels for boxes 

49. Dissecting tray 

50. White or light blue plastic sheet for 

photographic background 

51. 85% saline for relaxing parasites 

52. 10% NB Formalin 

53. Ethyl Alcohol 95% 

54. Isopropyl Alcohol 70% 

55. Glycerol 

56. Teflon container (3) 

57. Aluminum foil (1 roll) 

58. Knives 

59. Blank notecard

17 Jan/RBC 

HAWAIIAN MONK SEAL NECROPSY REPORT FORM 

NECROPSY #_________ (Assign Necropsy # in order of necropsies performed (only assign # if necropsy 

is actually performed). For example: if this was the second necropsy performed 

during the 2003 season, the Necropsy # would be 2). 

BACKGROUND INFORMATION: 

 

Seal ID No ___________________ 

Survival Factor Number _____________ 

Temporary ID Nos______________ 

 Island/atoll _________ Islet in 

atoll ______ 

Size/sex _________Age (if known) 

 Sector ________ Beach 

Position _________ 

Collect/record tag condition 

Temple tags ____________________ 

 Date/time found dead 

_________________ 

PIT tags read: Left _______________ 

 Date/time last seen alive 

_______________ 

 Right_______________ 

Body measurements (also record on Tag/Hand Card): 

Axillary girth_______________ cm 

Dorsal standard length _______ cm 

Ventral standard length _______ cm 

Total body mass ____________ kg 

Measurer ________________________ 

Died/euthanized/other (explain) _________ 

___________________________________ 

___________________________________ 

___________________________________ 

Ventral Standard Length

Dorsal Standard Length

Girth
 B-7 

Photographs taken? (if yes, add roll #s to 

Census Form) ________________________ 

____________________________________ 

 DATE/TIME OF NECROPSY________________RECORDER: ________________

ATTENDING DVM/ PERSON(S) PERFORMING NECROPSY:_______________ 

CLINICAL HISTORY: 

Reproductive status ___________________ Prior injuries/marks ________________ 

Last event seen, etc. 

____________________________________________________________________ 

____________________________________________________________________ 

____________________________________________________________________ 

PROBABLE CAUSE OF DEATH: 

Based on gross findings (field diagnosis) ___________________________________ 

Based on histopathology: _______________________________________________

ENVIRONMENTAL CONDITIONS (around the approximate time of death): 

Environmental factors (extreme weather conditions, i.e., dry/hot, storms, precipitation, 

temperature changes or other changes that may contribute to stress)__________________ 

________________________________________________________________________ 

 Ambient Temperature ________ Cloud Cover _______________ 

Water Temperature __________ Rainfall ___________________ 

Wind Speed ________________ 

 Wind Direction _____________ 

GENERAL CONDITION: 

Carcass Condition Code 

Code 1: just died

Code 2: fresh/carcass in good condition (rigor mortis, fresh smell, normal 

appearance, minimal drying of skin and mucous membranes, eyes clear, 

carcass not bloated, muscles and blubber firm, viscera intact and welldefined, guts with no gas) 

Code 3: fair/decomposed (carcass and organs intact, bloating, skin sloughing, mild 

odor, eyes sunken, dried mucous membranes, friable viscera, blubber oily, 

muscles soft but still intact, gut dilated with gas) 

Code 4: poor/advanced decomposition (carcass may be intact but collapsed, skin 

sloughing, strong odor, blubber soft with pockets of gas, liquified organs, 

blood thin and black, viscera friable difficult to dissect and easily torn, gut 

filled with gas) 

Code 5: mummified/skeletal remains (skin draped around bones, remaining 

tissues desiccated) 

Gas formation (run hands firmly over body to feel for gas): Y/ N 

Location:__________________ 

Maggots: Y/ N Maggot location:_____________________________________________ 
 B-8 

Nutritional state (fat/overweight; normal/average; thin/poor; starving/emaciated): ______ 

________________________________________________________________________ 

Hydration state (good; slightly dehydrated; moderately dehydrated; severely dehydrated; 

unknown): ______________________________________________________________ 

Pelage/fur coat (normal; fur loss; oil; molting; scruffy; parasites): __________________ 

Skin (normal; cracking; bleeding; dried; moist; smooth; rough; wounds; infection; 

masses; parasites; scars):___________________________________________________ 

Nails (normal; present; absent; torn; bleeding; cracked; crushed): __________________ 

Vibrissae/whiskers (normal; present; absent; torn):______________________________ 

Peripheral lymph nodes (palpable; obvious; unnoticeable; other): __________________ 

Examine all outside surfaces and note all anomalies. Photograph scars and other distinguishing 

Characteristics (see Necropsy Instructions section D): 
 B-9 

Abnormalities and markings (include bruises, wounds, contusions, old scars, condition of and tears of skin, 

external parasites). Copy scar card or make drawings as appropriate. 

DORSAL

VENTRAL

MALE

FEMALE
 B-10 

DESCRIBE Conditions of All Organs and Tissues and Any Lesions 

(Use The Descriptors Provided)! 

EXTERNAL EXAMINATION 

Eyes (Describe): 

Eyes should be collected first. Remove both eyes with eyelids and lacrimal gland. 

Eye, right - fix in formalin. Make a 2-3 cm cut in the globe along the interface of the sclera (white part) and 

the part normally in the eye socket. 

Eye, left - ultrafreeze if liquid nitrogen available; otherwise fix in formalin 

Attach sterile 18 g x 1.5" needle to sterile syringe and insert needle to collect aqueous humour from both 

eyes. 

Sp #____________ A Right Eye with Eyelid G (FM) Sp #________ Aqueous Humour G (LN) 

 B Left Eye with Eyelid G (LN OR FM)

Brain: 

Brain should be collected second only if the carcass condition is code 1 and it has been less than 1 hour 

since death has occurred. Expose the brain on fresh specimens only (Codes 1,2). Remove the top of the 

skull and cut away the soft tissues over the cranial vault. The procedure requires a good set of bone cutters 

and cleaver and/or ax/hatchet. Clip around the sides and cut dura mater (thin fibrous covering) attached to 

bone and then cut the tentorium cerebellum (thick connective tissue roof over cerebellum) with bone 

forceps and remove the skull cap. 

Remove INTACT. Handle gently and use sufficient formalin. Split right and left halves of brain using a 

new scalpel blade. Slice right half into 3 sections from front to back and fix for HP; slice left half into 1 cm 

wide sections from side to side (medial to lateral) and fix for HIP. Collect four 25 gm (lx2x2cm) sections 

from the left and right lobes and wrap in foil for toxicology analysis. 

Sp # ________ A Left Brain G (FM) C Left Brain 25 gm G (LN) G Right Brain 25gm G (LN) 

 B Right Brain G (FM) D Left Brain 25 gm G (LN) H Right Brain 25 gm G (LN) 

 E Left Brain 25 gm G (LN) I Right Brain 25 gm G (LN) 

 F Left Brain 25 gm G (LN) J Right Brain 25 gm G (LN) 

Skull: 

If the brain is not removed, save entire skull including mandible (flense and let sit outdoors in a container 

to be reduced by bugs). If the brain is removed, remove mandible (lower jaw) and all teeth within it and 

preserve as above. 

Sp #________ Entire Skull G OR Sp #________ Mandible G 

Mouth/Teeth (Describe conditions of mouth and teeth): 
 B-11 

Dental formula (Incisors (front teeth before canines) 2/2, Canines 1/1, Post-Canines (all teeth behind 

canines) 5/5 each side): 

I ______, C______, PC ____ each side

Describe nares: 

Insert sterile swab into nares, then place in cryovial. 

Sp # Nasal Swab G (LN)

Look for nasal mites, if observed, collect with sterile swab and place in vial filled with AG/FM. 

Sp #_______ Nasal Mites G (AG/FM) 

Describe penile opening/ vulva (circle appropriate orifice): 

Sp # Genital G (LN) 

Describe anus: 

Blubber Biopsy: 

The blubber biopsy is taken from the lateral aspect of the seal’s pelvic girdle, approximately 5-15 

centimeters cranial to the wing of the ileum. Gloves, scalpel/knife and sterile biopsy punch are to be used. 

One full thickness skin incision, through to the level of the blubber, 1-2 cm in length, and parallel to the 

long axis of the animal is made before the introduction of the biopsy punch. 

Through this incision, a sterile 6 mm biopsy punch is inserted, and a blubber sample is taken through the 

full depth of the blubber layer (2-5 cm). Use sterile or sanitized thumb forceps and/or scissors as necessary 

without damaging or contaminating blubber tissue. No glove contact! This blubber sample will be stored in 

a cryovial and frozen in liquid nitrogen for fatty acid radioactive isotope analysis. 

Sp#________ A Blubber Biopsy G (LN) 

Sp# ________ B Blubber Biopsy G (LN) 

INTERNAL EXAMINATION 

Do the internal examine in 4 phases: integument, head, thorax, and abdomen. Open each part separately to 

help prevent contamination and drying out of tissues. 

Blubber (measure at sternum or chest between the front flippers). 

Sternal blubber thickness: ______________ (mm) 

1. INTEGUMENT 

Hawaiian monk seal dermis and epidermis are fairly thick and dense. The deeper layers are firm and white. 

Immediately beneath this the blubber layer begins and for all practical purposes continues until the muscle 

begins. 
 B-12 

If a carcass is code 1 or 2, two sets of histopathology samples from the organs listed in the Necropsy 

Protocol should be collected. Both sets of HP samples will have the same specimen number, but two 

different sub-numbers as follows: 

Tissue Set Jar #1 = Sp #________ A 

Tissue Set Jar #2 = Sp #________ B 

Vibrissae:

When collecting vibrissae, include a 2 cm x 2 cm x l cm thick piece of tissue attached to vibrissae. 

Tissue Set Jar #1 

Left Vibrissae G (FM) 

Tissue Set Jar #2 

Right Vibrissae G (FM)

Flippers: 

Take small pieces of each flipper with nail and fix in formalin, as 2 sets. Also take a tissue plug from rear 

flipper and fix in 95% ethyl alcohol for genetics. 

Tissue Set Jar #1 

Front Flipper G (FM) 

Rear Flipper G (FM) 

Tissue Set Jar #2 

Front Flipper G (FM) 

Rear Flipper G (FM)

Sp #_______ Flipper (Tissue Plug for genetics) G (AA) 

2. HEAD 

Remove head from body and examine mouth, tongue, and collect lymphoid tissues and salivary glands. 

Lymph Nodes 

Submandibular with salivary glands (Describe): 

Tonsil (Describe): 

Tongue (Describe): 

Tissue Set Jar #1 

Submandibular/salivary gland G (FM) 

Tonsil G (FM) 

Tongue G (FM) 

Tissue Set Jar #2 

Submandibular/salivary gland G (FM) 

Tonsil G (FM) 

Tongue G (FM)

Unless carcass is a code 1 and the brain has already been removed, place head aside in a cool area and 

remove the brain last. 
 B-13 

3. THORAX 

Take 2 specimens including skin, subcutaneous tissue, and muscle for HP. Also collect 8 specimens of 

blubber without skin for toxicology (see FMPN p. 5) and 2 additional specimens of muscle only if animal 

was euthanized. 

Skin (Describe): 

Tissue Set Jar #1 

Skin G (FM)

Tissue Set Jar #2 

Skin G (FM)

Blubber (Describe): 

Tissue Set Jar #1 

Blubber G (FM) 

Tissue Set Jar #2 

Blubber G (FM)

Sp# __________ A Blubber 25 gm G

(LN) 

 B Blubber 25 gm G

(LN) 

 C Blubber 25 gm G

(LN) 

 D Blubber 25 gm G

(LN) 

E Blubber 25 gm G (LN)

F Blubber 25 gm G (LN) 

G Blubber 25 gm G (LN) 

H Blubber 25 gm G (LN)

Muscle (Describe): 

Tissue Set Jar #1 

Muscle G (FM) 

Tissue Set Jar #2 

Muscle G (FM)

Sp#________ A Muscle 1-100gm G (LN) 

Collect additional muscle sample if euthanasia: 

B Muscle 25 gm G (LN) 

C Muscle 25 gm G (LN) 

D Muscle 25 gm G (LN) 

E Muscle 25 gm G (LN) 

Brachial plexus (Describe): 

Prescapular/Axillary/Brachial plexus (nerve junction-line from thymus to shoulder, vessels and nerve tissue 

may be collected as one). Separate integument from rib cage and take nerve bundle. 

Tissue Set Jar #1 

Brachial plexus G (FM) 

Tissue Set Jar #2 

Brachial plexus G (FM)
 B-14 

Thyroid glands (Describe): 

Tissue Set Jar #1 

Left Thyroid G (FM) 

Tissue Set Jar #2 

Right Thyroid G (FM)

Open the chest and perform all collections within the chest. Do not open the abdomen yet. Examine and 

describe external surfaces of lungs and heart. 

After first opening the chest, use sterile syringe to extract chest fluid. Chest fluid will be present around the 

lungs or 

pooled along the diaphragm. 

Sp #_______ Chest Fluid G (LN) 

Thymus (Describe): 

Tissue Set Jar #1 

Thymus G (FM) 

Tissue Set Jar #2 

Thymus G (FM)

Heart (Describe): 

Attach sterile 18 g x 1.5" needle to sterile syringe and insert needle to collect pericardial fluid if present. 

Use same procedure with new sterile needle set up for whole blood collection from the heart. 

Sp #________ Pericardial Fluid G (LN) 

Sp #________ Whole Blood G (LN) 

Tissue Set Jar #1 

Right Atrium G (FM)

Left Atrium G (FM) 

Right Ventricle G (FM) 

Left Ventricle G (FM) 

Intraventricular Septum G (FM) 

Pulmonary Arteries G (FM) 

Aorta G (FM) 

Tissue Set Jar #2

Right Atrium G (FM)

Left Atrium G (FM) 

Right Ventricle G (FM) 

Left Ventricle G (FM) 

Intraventricular Septum G (FM) 

Pulmonary Arteries G (FM) 

Aorta G (FM)

Sp #________ A Heart G (LN) 

 B Heart G (LN) 

 C Heart G (LN) 

Lungs (Describe): 

 
 B-15 

Tissue Set Jar #1 Tissue Set Jar #2

Trachea G (FM) 

Bronchi G (FM) 

 Trachea G (FM) 

 Bronchi G (FM)

Right Lung G (FM) 

Left Lung G (FM) 

 Right Lung G (FM) 

 Left Lung G (FM)

Sp #_______ A Lung G (LN)

 B Lung G (LN) 

 C Lung G (LN)

Mediastinal Lymph Nodes (Describe): 

Tissue Set Jar #1 

Mediastinal lymph node G (FM) 

Tissue Set Jar #2 

Mediastinal lymph node G (FM)

Esophagus (Describe any abnormalities): 

Tie off both ends of esophagus (from the mouth-pharynx to within 1-2 cm of stomach) and place in bucket 

with formalin. When tying off the esophagus, first tie off both the distal end of one section and the 

proximal end of the next section about 1⁄2-1 inch apart then make the cut between the ties. 

Sp #________ Esophagus G (FM) 

4. ABDOMEN 

Liver (Describe): 

Collect liver tissues for histopathology (2 sets), toxicology samples (eight, 25 gm samples), biotoxin 

analysis (four, 25 gm samples), and for microbiology (two lxlxl cm samples). Collect bile from freshly 

dead seals only. 

Tissue Set Jar #1 

Liver G (FM) 

Tissue Set Jar #2 

Liver G (FM)

Sp #________ A Liver 25 gm G (LN) I Liver 25 gm G (LN) M Liver 1x1x1cm

G (LN) 
 B-16 

 B Liver 25 gm G (LN) J Liver 25 gm G (LN) N Liver 1x1x1cm

G (LN) 

 C Liver 25 gm G (LN) K Liver 25 gm G (LN)

 D Liver 25 gm G (LN) L Liver 25 gm G (LN) 

 E Liver 25 gm G (LN) 

 F Liver 25 gm G (LN) 

 G Liver 25 gm G (LN) 

 H Liver 25 gm G (LN) 

 

Sp #________ Bile G (LN) 

Pancreas (describe):

Tissue Set Jar #1 

Pancreas G (FM) 

Tissue Set Jar #2 

Pancreas G (FM)

Spleen (Describe): 

Collect tissues for HP (2 sets) and for microbiology (two lxlxlcm samples) if liquid nitrogen available. 

Tissue Set Jar #1 

Spleen G (FM) 

Tissue Set Jar #2 

Spleen G (FM)

Sp# __________ A Spleen C (lxlxl cm) G (LN) 

B Spleen D (lxlxl cm) G (LN) 

Gastro-intestinal Tract/Alimentary Canal: 

Having already tied off the distal end of the esophagus in part 3, examine serosa of tract from ties to anus. 

When removing GI tract dissect intestines from mesentery, check mesentery for parasites, and surface of 

intestines for nodules, vesicles, segmented discoloration, or adhesions. Color can be normal, green, 

congested, purple, black, red, yellow, white, for example; contents can be described as empty, bile, digesta, 

for example. Unless indicated by severe decomposition or suspect condition, GI tract may be removed in its 

entirety. 

If specimen is too large for a 5-gallon bucket, it can be subdivided as follows: 

1. Stomach: ligate (tie-oft) at both ends. 

2. Intestine-rectum-anus: ligate at both ends. 

For ALL necropsies, measure the length of the various components of the GI tract, including esophagus, 

stomach, and intestines to rectum. This can be done on fixed tissues later or in the lab. This does not need 

to be done on fresh tissues in the field unless you have lots of free time. 

Measurements: 

Mouth (Pharynx) to anus ___________________ 

m 

Greater curvature of stomach 
 B-17 

________________ cm 

Lesser curvature of 

stomach_________________ cm 

Distal sphincter (end) of stomach to anus 

________m 

Cecum to anus _________________________ 

cm 

(Cecum is blind pouch at junction of sm and lg 

intestine)

GIT (Describe): 

Sp # GIT G (FM) 

Mesenteric Lymph Nodes (Describe): 

Tissue Set Jar #1 Tissue Set Jar #2 

Mesenteric Lymph Node G (FM) Mesenteric Lymph Node G (FM)

Peritoneal Fluid (Describe): 

Collect approximately 10 ml of peritoneal fluid with sterile syringe and store in cryovial. 

Sp # Peritoneal Fluid G (LN) 

Urogenital System: 

Examine and describe kidney, adrenal glands, and reproductive tract. 

Kidney (Describe): 

Include cortex when collecting kidney. Collect tissues for HP (2 sets), eight, 25 gm samples for toxicology, 

and two lxlxl cm samples for microbiology if liquid nitrogen available. 

Tissue Set Jar #1 

Left Kidney G (FM) 

 Tissue Set Jar #2 

 Right Kidney G (FM)

Sp #_______ A Kidney 25 gm G (LN) E Kidney 25 gm G (LN) I Kidney 1x1x1cm 

G (LN) 

 B Kidney 25 gm G (LN) F Kidney 25 gm G (LN) J Kidney 1x1x1cm 

G (LN) 

 C Kidney 25 gm G (LN) G Kidney 25 gm G (LN)

 D Kidney 25 gm G (LN) H Kidney 25 gm G (LN) 

Adrenal (Describe): 

Tissue Set Jar #1 Left Adrenal G (FM) 
 B-18 

Tissue Set Jar #2

Right Adrenal G (FM) 

Urinary bladder (Describe): 

The urinary bladder will normally be found attached to the ventral wall of the abdomen. The urinary 

bladder can be empty, dilated, or thickened. Examine for tumors. 

Attach a sterile 18 g x l.5" needle to a sterile syringe, insert the needle into bladder, and draw on 

syringe to collect urine. 

Urine: amount: _______ cc color: ________ consistency:____________ 

Sp #_________ Urine G (LN) 

Tissue Set Jar #1 

Urinary Bladder G (FM) 

Tissue Set Jar #2 

Urinary Bladder G (FM)

Reproductive Tract: 

MALES (For all males, examine, describe, and sample reproductive tract): 

If both testes appear grossly normal, collect a 1 cm thick cross section of both testes for each HP jar and a 1 

cm thick cross section of both testes for toxicology. 

Tissue Set Jar #1 

Left Testes G (FM) 

Right Testes G (FM)

Tissue Set Jar #2

Left Testes G (FM) 

Right Testes G (FM) 

Sp #__________ A Left Testes G (LN) 

 B Right Testes G (LN) 

FEMALES (Externally examine the vestibule and surrounding area and describe 

observations): 

If lactating, collect as much milk as possible and freeze. Collect double HP specimens from each of the 

four teats and fix in formalin. 

Milk (Describe): 

Sp Milk G (LN) 
 B-19 

Mammary Glands (Describe): 

Tissue Set Jar #1 

Mammary Gland 1 G (FM) 

Mammary Gland 2 G (FM) 

Mammary Gland 3 G (FM) 

Mammary Gland 4 G (FM) 

Tissue Set Jar #2

Mammary Gland 1 G (FM) 

Mammary Gland 2 G (FM) 

Mammary Gland 3 G (FM) 

Mammary Gland 4 G (FM)

Reproductive Tract: 

Collect whole reproductive tracts from all females, including immature seals and pups. Include the 

external genitalia, ovaries, uterus and cervix; measure ovaries; and fix in formalin. Do not slice open 

the ovaries (they are small enough that they will fix completely), but note any discolorations, ruptured 

follicles, etc. Cut several cuts 

into the body of the uterus to allow formalin to flow into the lumen for better fixation of endometrium. 

Note presence of fetus, abscesses, tumors, unusual amounts of fluid, etc. Examine ovaries for symmetry, 

scars, cysts, or other structures "normal" or "abnormal." Measure ovarian structures.

Sp # Female Reproductive Tract G (FM) 

Ovaries (Describe): 

Measurements: Left Ovary_____ cm x ______cm 

 Right Ovary _______cm x _______cm 

For pregnant females, aborted fetuses, or perinatal pup deaths, 

examine and sample (or collect) umbilicus, placenta and 

fetus. 

Umbilicus (Describe): 

Tissue Set Jar #1 

Umbilicus G (FM) 

Tissue Set Jar #2 

Umbilicus G (FM)
 B-20 

Placenta (Describe): 

Handle tissues in a sterile manner. Fix samples for histopathology as follows: remove four 5 cm x 1cm full 

thickness strips (extending through to include both the fetal and maternal side) representative of normal 

and any abnormal portions of the placenta. If freezing is an option, in addition to preserving sections for 

histopathology, freeze four 5 cm x 1 cm full thickness strips (extending through to include both maternal 

side) representative of normal and any abnormal portions of the placenta. If freezing in liquid nitrogen, 

BE SURE samples are small enough to be removed from the dewar. If this is not possible, but 

immediate transportation is available, send to Honolulu on ice. DO NOT STORE TISSUES WITH FOOD 

FOR HUMAN CONSUMPTION! 

Tissue Set Jar #1 

Placenta G (FM) 

Placenta G (FM) 

Tissue Set Jar #2 

Placenta G (FM) 

Placenta G (FM) 

Sp # _________ A Placenta G (LN) 

 B Placenta G (LN) 

 C Placenta G (LN) 

 D Placenta G (LN) 

Fetus (Describe): 

If fetus is <25 cm in length, split it in half from chin to pubis and place both halves in formalin. If fetus is 

>25 cm in length, perform complete necropsy. 

If complete necropsy is performed, use separate Necropsy Report Form. Handle tissues in a 

sterile manner. 

Take one swab from both the throat and rectum before beginning necropsy. 

If fetus is fresh, collect several cc’s of fluid from the stomach and freeze in liquid nitrogen. 

 Ventral Length: ________ cm 
 B-21 

 Axillary Girth: ________ cm 

 Mass: ________ kg 

Sex: M or F 

Condition (Describe): 

_____________________________________________________________ 

Fetus Necropsied: Y or N (if Y, see Necropsy # ________)

If Fetus is < 25 cm: OR If Fetus is > 25 cm: 

Sp #__________ A Fetus G (FM) Sp #__________ Throat Swab G (LN) 

 B Fetus G (FM) Sp #__________ Rectal Swab G (LN) 

 Sp #__________ Stomach Fluid G (LN) 

25 Feb 2003/RCB 
 B-22 

NECROPSY SPECIMEN REFERENCE TABLE (Listed by order of collection):

Tissue Diagnostic Activity Size of Sample

No. 

Samples Preservation Notes

Eye (right), with 

eyelid and 

lacrimal duct 

Histopathology Whole eye 1 Make a 2-3 cm cut in the 

glove along the interface of 

the sclera (white part) and the 

part normally in the eye 

socket 

Fix both eyes in 

FM if freezing not 

possible 

Eye (left), with 

eyelid and 

lacrimal duct 

PCR/Microscopy Whole eye 1 Whirlpak in liquid nitrogen 

Aqueous humour PCR/Micro As much as 

possible 

Both eyes Cryovial, in liquid nitrogen Collect fluid with 

sterile needle/ 

syringe 

Brain, left half Histopathology Slice into 1 cm 

wide sections 

1 10% formalin Slice from side to 

side (medial to 

lateral) 

Brain, right half Histopathology Slice into 3 

sections 

1 10% formalin Slice from front to 

back 

Brain Toxicology 25 g 8 Wrap in foil, shiny side out, 

in liquid nitrogen 

Skull Aging Whole 1 Dry Flense outdoors 

Mandible Aging Lower jaw 1 Dry Flense outdoors 

Nasal swab Virology 1 Cryovial, in liquid nitrogen 

Nasal mites Parasitology 1 AG or 10% formalin If nasal mites 

present, collect 

with swab 

Genital swab Virology 1 Cryovial, in liquid nitrogen 

Blubber Biopsy Fatty Acid Analysis 6 mm 1 5 ml cryovial, in liquid 

nitrogen 

See blubber 

biopsy protocol 

Blubber Biopsy Toxicology 6 mm 3-4 plugs Teflon container, in liquid 

nitrogen 

See blubber 

biopsy protocol 

Vibrissae Histopathology 5 cm X 5 cm X 

1 cm 

2 Include 2x 2 x 1 cm thick 

tissue attached to vibrissae 

Flipper, front, w/ 

nail 

Histopathology 2 10% formalin 

Flipper, rear Histopathology 2 10% formalin 

Flipper, rear Genetics 1 95% ethyl alcohol 

Lymph nodes, 

submandibular 

with salivary 

gland 

Histopathology 3 cm X 3 cm X 

1 cm 

2 10% formalin 
 B-23 

Tissue Diagnostic Activity Size of Sample

No. 

Samples Preservation Notes

Tonsil Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin 

Tongue Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin 

Skin Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin 

Blubber Histopathology 1 cm X 3 cm X 

3 cm 

2 10% formalin Sample includes 

skin, subcutaneous tissue 

and muscle 

Blubber Toxicology 25 gm 8 Wrap in foil, shiny side out, 

in liquid nitrogen 

Sample from 

sternal region, 

without skin 

Muscle Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin 

Muscle, skeletal Biotoxins 1-100 g 1 Whirlpak, in liquid nitrogen 

Muscle, skeletal Toxicology 25 gm 4 Wrap in foil, shiny side out, 

in liquid nitrogen 

Collect if 

euthanasia only 

Brachial plexus Histopathology All one side 2 10% formalin 

Thyroid Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin 

Chest Fluid Pathology 10 ml 1 Cryovial, in liquid nitrogen 

Thymus Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin 

Pericardial fluid Pathology As much as 

possible 

1 Cryovial, in liquid nitrogen 

Whole blood Biotoxin 1-100 ml 1 Teflon container, in liquid 

nitrogen 

Heart, aorta Histopathology 3 cm section 2 10% formalin 

Heart, 

intraventricular 

septum 

Histopathology 3 cm X 3 cm X 

1 cm 

2 10% formalin 

Heart, right atrium Histopathology 3 cm X 3 cm X 

1 cm 

2 10% formalin 

Heart, left atrium Histopathology 3 cm X 3 cm 

X 1 cm 

2 10% formalin 

Heart, left 

ventricle 

Histopathology 3 cm X 3 cm X 

1 cm 

2 10% formalin 

Pulmonary 

arteries 

Histopathology 3 cm X 3 cm X 

1 cm 

2 10% formalin 

Heart Biotoxins 1-100 g 3 Whirlpak in liquid nitrogen 
 B-24 

Tissue Diagnostic Activity Size of Sample

No. 

Samples Preservation Notes

Trachea Histopathology 5cm X 5 cm X 

1 cm 

2 10% formalin 

Bronchi Histopathology 3 cm long 

section 

2 10% formalin 

Lung, right Histopathology 5 x 5 x 2.5 cm 2 10% formalin 

Lung, left Histopathology 5 x 5 x 2.5 cm 2 10% formalin 

Lung PCR/Micro 1 x 1 x 1 cm 3 Whirlpak in liquid nitrogen 

Lymph nodes, 

mediastinal 

Histopathology 3 cm X 3 cm X 

1 cm 

2 10% formalin 

Esophagus Histopathology 1 10% formalin 

Liver Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin 

Liver Toxicology 25 g 8 Wrap in foil, shiny side out, 

in liquid nitrogen 

Slice from caudal 

end of left anterior 

lobe; collect 8 

samples, 12 if 

euthanasia; 

Liver Biotoxins 25 g 4 Whirlpak in liquid nitrogen 

Liver PCR/Micro 1x1x1 cm 2 Whirlpak in liquid nitrogen 

Bile Toxicology No more than 

12 ml 

1 Cryovial, in liquid nitrogen Fresh dead only 

(codes 1,2) 

Pancreas Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin 

Spleen Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin 

Spleen PCR/Micro 1x1x1 cm 2 Whirlpak in liquid nitrogen 

GIT Histopathology Whole GIT 1 10% formalin 

Lymph nodes, 

mesenteric 

Histopathology 3 cm X 3 cm X 

1 cm 

2 10% formalin 

Peritoneal fluid Microbiology No more than 

12 ml 

1 Cryovial, in liquid nitrogen 

Kidney Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin Include cortex 

Kidney Toxicology 25 g 8 Wrap in foil, shiny side out, 

in liquid nitrogen 

Take sample from 

caudal end of each 

kidney; include 

cortex 

 
 B-25 

Tissue Diagnostic Activity Size of Sample

No. 

Samples Preservation Notes

Kidney PCR/Micro 1x1x1 cm 2 Whirlpak in liquid nitrogen 

Adrenal Histopathology Both 2 10% formalin 

Urine Toxicology No more than 

12 ml 

1 Cryovial, in liquid nitrogen 

Urinary bladder Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin 

Testes Histopathology 1cm thick cross 

section of both 

2 10% formalin 

Testes Toxicology 1cm thick cross 

section of both 

2 Wrap in foil, shiny side out, 

in liquid nitrogen 

Reproductive 

tract, female 

Histopathology Entire tract 

whole 

1 10% formalin 

Milk Nutritional analysis As much as 

possible 

1 Cryovial, in liquid nitrogen 

Mammary glands Histopathology 5 cm X 5 cm X 

1 cm 

8 10% formalin 2 sets of 1 each 

Umbilicus Histopathology 5 cm X 5 cm X 

1 cm 

2 10% formalin 

Placenta Histopathology 5 cm long x 1 

cm wide 

4 10% formalin Include both fetal 

and maternal sides 

Placenta Microbiology 5 cm long x 1 

cm wide 

4 Whirlpak in liquid nitrogen Include both fetal 

and maternal sides 

Fetus See notes See notes See notes See notes Complete 

necropsy or split 

in 2 and preserve 

in formalin 

Fetal Throat Swab Bacteriology 1 Cryovial, in liquid nitrogen 

Fetal Rectal Swab Bacteriology 1 Cryovial, in liquid nitrogen 

Fetal Stomach 

Fluid 

Microbiology Several 

ccs 

Cryovial, in liquid nitrogen 

Lesions Cytology Slides 2 1 fixed in 95% ethyl alcohol, 

1 air dried 

Take 2 smears 

from any lesion 

Lesions Histology Include the 

margin 

between the 

abnormal and 

adjacent 

normal tissue 

2 10% Formalin 

 

APPENDIX C: 

Equipment and Supplies for a Single Necropsy 

 C- 1

28 Jan2003/RCB 

EQUIPMENT AND SUPPLIES FOR A SINGLE NECROPSY 

1. Clipboard, metal 

2. Necropsy Table (Checklist) 

3. Necropsy Report Form(s) 

4. Necropsy Manual 

5. Scarcard 

6. Census Form 

7. Tag/handling card 

8. Survival factor form 

9. Self-sticking pre-printed specimen labels 

(3 sheets) 

10. Waterproof labels ("rite-in-rain" paper, 

pre-cut) (3 sheets) 

11. Pencils (2) 

12. Ball-point pens (optional) (2) 

13. Permanent markers (sharpies) (2) 

14. Ruler (metric) 

15. Metal tape measure (metric) 

16. Tape measure, flexible (metric) 

17. Surgical scissors (2: straight and curved) 

18. Rubber gloves 

chemical resistant - 2 pr. latex 

heavy duty - 6 pr. latex exam 

gloves - 15 pr 

19. Scalpel handles (4: 2 #8, 2 #6) 

20. Scalpel blades (30) 

21. Blubber biopsy punch, 6 mm (2) 

22. Forceps (2 pr.) 

23. Hemostats (1) 

24. Dacron Swabs (5) 

25. Microscope slides 

26. Slide holder for microscope slides 

27. Small vials (—2 x 1⁄2") - 6 ea. 

28. Med. vials (—3' x 2) - 6 ca. 

29. Whirlpak bags (20 ea.) 

30. Ziploc bags (large) 

31. Paper towels 

32. Antibacterial soap/Hand sanitizer 

33. Sharps container 

34. Needles, 1 1⁄2', 18g (5) 

35. Syringes, 20cc or 35cc (5) 

36. Blood tubes, PT, TT (4) 

37. Biohazard bag (2) 

38. Camera with colored slide film and/or 

Digital camera 

39. Shovels 

40. Jars and buckets partially filled with 10% 

NB Formalin 

a. 2 large jars for duplicate 

sets of histopathology 

tissue samples 

b. 1 bucket for entire female 

reproductive tract 

c. 1 bucket for esophagus 

d. 1 5gal. bucket for entire 

gastrointestinal tract from 

beginning of stomach to 

anus 

e. 2 jars for eyes 

f. 2 jars for brain 

41. Hacksaw 

42. Axe 

43. Cryovials (20) 

44. String to tie off beginning of 

stomach, etc. (1 roll) 

45. Cutting board (not on spc. coil. 

list) 

46. Disposable aprons (3) 

47. Bone shears 

48. Archival tag labels for boxes 

49. Dissecting tray 

50. White or light blue plastic sheet for 

photographic background 

51. 85% saline for relaxing parasites 

52. 10% NB Formalin 

53. Ethyl Alcohol 95% 

54. Isopropyl Alcohol 70% 

55. Glycerol 

56. Teflon container (3) 

57. Aluminum foil (1 roll) 

58. Knives 

59. Blank notecard

APPENDIX D: 

Hawaiian Monk Seal Rescue and Rehabilitation Guidelines

 D - 1

HAWAIIAN MONK SEAL RESCUE AND REHABILITATION GUIDELINES

 

I. QUARANTINE

 

A. QUARANTINE DEFINITION AND OBJECTIVES 

1. Quarantine refers to isolation or restriction on travel or passage imposed to 

keep contagious diseases, insect pests, etc. from spreading. Since 1989, NMFS 

has maintained quarantine policies within the rehabilitation program. 

2. Hawaiian monk seals held at Kewalo Research Facility (KRF) must be 

maintained under strict quarantine at all times to a) minimize transmission of 

disease from outside sources, b) minimize the spread of disease from these seals 

to susceptible animals on Oahu, including humans, and c) minimize transmission 

of disease among the three holding tanks at KRF. 

3. All personnel involved in the feeding, handling, and care of these seals must be 

properly trained in quarantine procedures by an experienced NMFS or the Joint 

Institute for Marine and Atmospheric Research (JIMAR) staff. (Quarantine 

procedures should always be posted in the food preparation area.) 

B. NMFS QUARANTINE POLICY 

1. All equipment used in the quarantine facility, including feeding, handling, and 

medical supplies MUST be A) labeled with “MONK SEAL QUARANTINE”

and its specific use B) used exclusively for quarantined seals C) kept separate 

from equipment used for other animals (wild monk seals, dolphins, etc) and D) 

properly disinfected before and after entering the quarantine enclosures as 

dictated by Dr. Bob Braun. 

2. Due to the unknown risk of infection from these seals, feeding, handling, and 

medical procedures involving non-quarantined animals should precede contact 

with the KRF seals whenever possible. 

3. Avoid direct contact with domestic or other captive or wild animals before and 

after entering quarantine enclosure. 

4. Shower and change clothes before and/or after going to another animal care 

facility if entering the KRF seal enclosures on the same day. 

5. Wear rubber boots designated for monk seal quarantine use at KRF in the 

enclosures at all times. Minimize wearing quarantined boots around KRF 

premises. Separate quarantined boots are provided for tank 3. No street shoes are 

to be worn into the enclosures, including the walkway leading to tanks 1 and 2. 
 D-2 

6. Dip soles of boots in disinfectant footbaths bath upon entering AND leaving 

all the enclosures. 

 

7. Immediately upon entering the enclosure to tanks 1 and 2, wash hands with 

antibacterial soap. Also wash hands before and after fish preparation, feeding, or 

handling seals. Always wash hands immediately after leaving a separate tank 

enclosure. Before and after entering tank 3, wash hands at the yellow sink next to 

the enclosure. 

8. Any person who will potentially come into direct contact with quarantined 

seals for any procedure must wear sanitary protective clothing (i.e., coveralls) 

designated for quarantine monk seal use only. This clothing should be kept clean 

and in a designated area (in the monk seal lab) away from potential sources of 

contamination. 

9. Restrainers and medical staff must shower after animal handling procedures. 

Protective clothing worn during procedures should be immediately washed in the 

KRF washer with soap and dilute bleach solution. Do not set this clothing on any 

surface outside of quarantine enclosures prior to washing. 

II. OBSERVATIONS AND CONDUCT AROUND SEALS

A. INITIAL INSPECTION OF SEALS 

1. Prior to feeding, conduct a brief inspection of the seals and tank enclosures. 

2. Keys to seal enclosures are kept inside first shelf in MMRP cage (combination 

#0723). 

3. Try to avoid alerting the seals to your presence. Observe condition of seals, 

water levels, feces (scat), urine, regurgitation (spew), and harmful debris on haulout areas and on bottom of tanks. When possible, note ID of seal that produced 

scat, etc. 

4. Note anything unusual in a seal’s normal appearance (eyes, nasal discharge, 

bite wounds, etc.) and behavior (lethargic, unresponsive, etc.). Notify attending 

veterinarian and head caretaker immediately of any abnormal changes in a seal's 

health. 

5. Succinctly record any observations on health record forms (refer to 

"Instructions for Completing the Hawaiian Monk Seal Feeding and Health Record 

Form"). 
 D-3 

B. CONDUCT AROUND SEALS AT ALL TIMES 

1. When in enclosures, DO NOT MAKE PHYSICAL CONTACT WITH 

SEALS unless necessary for procedures requiring handling. 

2. If seals are resting or sleeping, do not make loud noises or startling gestures, 

and move slowly when in close proximity to them to minimize stress. 

3. Whenever possible, observers should remain as inconspicuous and unobtrusive 

as possible to observe seals’ normal behaviors in captivity and minimize their 

stress in captivity. 

4. Seal aggression displayed during feedings should be controlled through the 

application of behavior modification techniques. Specific aggressive and 

undesirable behaviors that are subject to behavior modification include: a) 

approaching too closely or lunging at the feeder during feedings; b) splashing, 

sharp movements, open mouth displays, or loud vocalizations which appear as 

demands for fish; c) approaches toward or contact with the fish bucket; and d) 

displays of inter-seal aggression. Immediately after the occurrence of any of 

these behaviors feeders should avoid administering a fish. If the behavior persists 

or escalates impose a "time out" by temporarily leaving the enclosure with the 

fish bucket. As a general rule do not administer fish immediately after the 

occurrence of any undesirable behavior, and attempt to present fish when the seals 

are orderly, well stationed, and nonaggressive. 

5. Outside of feeding sessions seals may display undesirable behaviors which 

include: a) approaching too closely or too rapidly; b) mouthing hoses, brooms, or 

boots; and c) stereotypic behaviors which include repetitive splashing or slapping 

at the walls of the enclosure. If seals approach too closely or too rapidly, use the 

broom quickly to push the seal away but avoid getting confrontational with seals 

or allowing the seal to mouth the broom. The mouthing of brooms, hoses, and 

boots should be discouraged by preventing opportunities for seals to bite at these 

objects in the first place. Stereotypical behaviors are a sign of boredom and may 

be reduced by providing seals with their approved environmental enrichment 

devices (buoys and PVC pipe toys). 

III. SEAL CARE

A. FISH PREPARATION

1. Review posted food amounts before preparing fish to note any changes in 

feeding amounts. 

2. Wash hands thoroughly with antibacterial soap before handling buckets and 

fish. 
 D-4 

3. Use buckets hanging in fish house labeled for each tank enclosure. Do not 

allow unsanitized objects to come in contact with insides of buckets, and 

minimize such contact with their bottoms. (Dip bottoms of buckets in footbath 

before feeding if necessary.) 

4. Weigh fish from the hanging, digital metric scale. Turn scale “ON,” and allow 

it to zero. Hang bucket, let scale equilibrate, then push the “ZERO” button. After 

scale has equilibrated to zero again, begin weighing fish. Zero scale for each 

bucket. 

 

5. Herring is stored in boxes and plastic bags in the freezers marked "MONK 

SEAL FOOD ONLY." ALL FISH FOOD MUST HAVE A DOUBLE-LAYER 

OF PROTECTION AND BE WELL-SEALED TO PREVENT FREEZER 

BURN AND DRYING. Always tie plastic bags of herring after weighing fish.

6. FREEZERS AND REFRIGERATORS MUST REMAIN CLEAN AND 

NEAT AT ALL TIMES. All feeders are responsible for maintaining cleanliness 

of the food preparation area on a daily basis. 

7. While weighing fish, inspect and remove any damaged fish (e.g., no head, 

bruised). Store these fish in a plastic bag in freezer for use by MTRP. Do not 

place any trash with damaged fish. 

8. Thaw fish in refrigerator. 

9. As fish are thawing, inspect again for bruises, cuts, or other abnormalities not 

detectable when frozen. Replace any unacceptable fish. 

10. Promptly feed first tank while fish are cold. Place any other thawed fish in 

refrigerator until ready to feed. 

11. Prepare vitamins for each seal in the morning (type and amount per the 

attending veterinarian’s directions). Place vitamins in one fish per seal, inserted 

behind gills and into body cavity. Pull off a pectoral fin or cut off a small portion 

of the tail to identify this fish and ensure it is ingested. If a seal normally guts 

fish, insert vitamins into cut tail, one per fish. If morning vitamins are missed, 

leave a note that they should be given in the evening. 

B. FREE FEEDING 

 

1. WASH HANDS WITH ANTIBACTERIAL SOAP IMMEDIATELY PRIOR 

TO AND AFTER FEEDING. 

2. DIP BOOTS IN FOOTBATH UPON ENTERING AND LEAVING SEAL 

ENCLOSURES. If bottoms of buckets have touched unsanitized areas, dip them 

in footbath before entering. 
 D-5 

3. Maintain a safe distance from seals at all times. If seals approach gate too 

closely to allow entrance, use a quarantine broom to keep them away; or throw a 

fish into water before entering enclosure. 

4. ALWAYS KEEP DOOR SECURELY CLOSED WHILE IN SEAL 

ENCLOSURES. 

5. Give pilled fish after observing that fish are being eaten whole (third or fourth 

fish). Closely observe if pills are ingested or sink to bottom. If a seal does not 

receive morning vitamins, record this on the feeding record and leave a note for 

the evening feeder. Note if each seal appears satiated or is still hungry after 

feeding, and if any unusual behavior or clinical symptoms arise. 

6. While feeding, keep seals as far away from feeder and from each other as 

possible. 

7. Feed at the same pace without letting one seal steal from another. If this 

happens, replace stolen fish with fish from stealer. Try not to throw another fish 

until all seals have completely ingested their fish. 

8. Never allow seals to take fish directly from buckets. 

9. When finished feeding, set buckets outside of enclosure and thoroughly clean 

deck. While cleaning deck, observe seals for any regurgitation, urination, 

defecation, and presence or absence of worms in feces. 

 10. If any uneaten fish remain in bucket, weigh and store in trash bag in freezer 

 for discard (not for MTRP). Record how much each seal actually ate, estimating 

 weight of additional fish ingested or lost fish when necessary. 

C. DAILY CLEANING

1. Clean scat, urine, fish parts, and wind-blown debris from the decks and walls 

of each enclosure prior to feeding seals. Use the brooms and fresh water hoses 

that are specifically designated for the cleaning of each seal enclosure. 

2. Do not allow seals to mouth or bite brooms or fresh water hoses. If the hose 

enters the tank remove it immediately. Bolts have been installed on the railings of 

each enclosure to hang hoses out of the seals’ reach during cleaning. 

3. Never allow the broom, hose, or any equipment to remain unattended in a seal 

enclosure. Return all equipment to its storage area (i.e., coil and hang hose, place 

broom in Nolvasan bath) area after use. 
 D-6 

4. When cleaning, take the opportunity to inspect urine for color, and feces for 

consistency and parasites. Always record feces and urine in the feeding log and 

make special note of any unusual findings. 

5. As an aid to cleaning enclosure surfaces the decks may be flooded with sea 

water by temporarily closing the outlet valves. If the outlets are closed for 

cleaning, be sure to re-open them immediately after cleaning is complete and 

before feeding begins. Avoid feeding the seals when the enclosure is flooded 

because this will distribute scales and fish debris over the surface of the decks. 

6. Always keep the enclosure doors securely bolted because the seals are very 

adept at exiting the enclosures through a door left ajar. 

7. After feeding, thoroughly rinse all fish scales, blood, and debris from the decks 

and walls of the enclosure. Special care should be taken to clean scales from 

doors, door handles, and bolts. 

8. The buckets may be rinsed in the sea water inflow but do not allow the seals to 

touch, mouth, or bite at the buckets. Rinse buckets thoroughly in the fish house 

sink, then scrub them with scouring powder and a scrubby (inside bottom first, 

then inside walls, then outside). Rinse the buckets well and hang to dry with lip 

facing down. 

9. The sink and any feeding-related equipment (rubber gloves, cutting board, 

plastic containers, scrubbies, sink screens etc.) should be cleaned with chlorinated 

scouring powder after each feeding and rinsed well. 

10. Clean any fish scales, blood, or debris from the fish house after each feeding. 

Mop the fish house floor with a 20% bleach solution after the afternoon feeding. 

11. The common walkway to enclosures 1 and 2 should be scrubbed and rinsed 

after each feeding. 

 12. Footbaths are changed every 2 days or as needed using 6 tablespoons (3 oz.) 

 of Nolvasan per gallon of water or designated measuring cup attached to each 

 bottle. 

13. Check seals one last time before leaving. Ensure gate to enclosure is locked, 

water hose is turned off, and drain valve is at appropriate setting. If water to tank 

was turned off/on, recheck main source. NEVER LEAVE IN A HURRY; 

always double check that all tasks are completed. 

14. Return key to seal enclosure to cage and securely lock. If no one else is on 

premises, make sure all doors and gates are securely locked. 
 D-7 

D. WEEKLY CLEANING 

1. The monk seal tanks and enclosures are drained and cleaned once a week. A 

minimum of three people should conduct the weekly cleaning. 

2. At the start of cleaning, change all foot and broom baths to a 20% bleach 

solution. Thoroughly rinse all Nolvasan solution from the brooms, broom baths, 

and footbaths to avoid Nolvasan/bleach reactivity and staining. 

3. Any personnel who will be responsible for draining the tanks must receive 

specific training. The tanks drain in approximately 30 minutes and during this 

time the seals must be monitored to ensure that they remain in their tank as it is 

lowered. Care must be exercised to assure that a seal is not stranded on the deck 

of a drained pool. 

4. While the tank is draining, scrub the entire upper deck and walls of the 

enclosure with a 20% bleach solution. Let the bleach stand for 10 minutes, then 

thoroughly rinse with fresh water. When using bleach solutions always direct the 

rinse water away from seals because the bleach solution is a skin and eye irritant. 

5. Sanitize the fiberglass partitions and place them inside the tank to isolate seals 

from the portion of the tank being cleaned. Broom the walls and floor of the tank 

with 20% bleach solution. Allow the solution to stand for 10 minutes and than 

thoroughly rinse with fresh water. Goggles and masks are available and should 

be used when cleaning the tank with bleach. 

6. Do not clean more than one half of the tank at a time and always avoid seal 

exposure to the bleach solution. Keep the seals cool by spraying with fresh water 

and monitor the seals’ behavior and status regularly. 

7. After the entire tank has been cleaned and rinsed, remove all cleaning 

equipment and turn on the sea water inflow. 

8. Scrub the common walkway to tanks 1 and 2 with a 20% bleach solution. 

9. After all the tanks and walkways have been cleaned, thoroughly rinse the 

bleach solution from the brooms, broom baths, and footbaths. Replace the 

Nolvasan solution (3 oz/1 gal) to all broom and footbaths. 

IV. PROCURING SEAL SUPPLIES 

A. HERRING & CAPELIN SUPPLY 

1. Each week, assess the amount of herring (in cases) that can be stored in the 

freezers at KRF. 
 D-8 

2. Call "Unicold Corporation" (Phone 836-2931) one day in advance and arrange 

a time for the pickup of a specific amount of herring (in cases). 

3. Drive to "Unicold" (address: 3140 Ualena Street, Honolulu, HI 96819) and 

pick up the herring 

4. Store the herring cases in the freezers at KRF making sure that any old stock is 

moved to the fish house freezer first. Always store new cases on the bottom of 

the freezer and old cases on top. 

5. After delivering the herring to KRF, turn in the invoice from “Unicold” along 

with a pinksheet to Administrative Support Assistant Bonnie Oshiro in the 

Administrative Office. PIFSC has a blanket purchase order (BPA) with 

“Unicold” so be sure to notate that it is a BPA charge on the pinksheet before 

submitting it to Bonnie. If the invoice cannot be taken to Bonnie on the day of 

pick-up, e-mail her a message indicating the date and the amount of herring 

picked up. 

 6. Empty herring boxes are folded and thrown in the dumpster. 

B. VITAMINS 

1. Vendor varies. Refer to attending veterinarian for type and amount.

V. WATER QUALITY

1. Water sampling should occur on the same day (Tuesday) and time (1000, no 

later than 1430) each week, the day before tank cleaning. Sample between 2700

and 3600 clockwise with water inlet at 3600. 

2. Do not open lid to bottle until immediately before collection; do not 

contaminate inside of lid or bottle. 

3. Place bottle countercurrent and dip underwater approximately 1 foot. Move 

bottle underwater countercurrent for a few feet and make sure it is at "fill" line, 

replace lid, and remove from water. 

4. Label bottle with the following information: 

a. Source of sample: Kewalo monk seal tank 

b. Collected by: NMFS/initials of collector 

c. Date and time (Tuesday mornings) 

d. Tests required: Fecal coliform 

5. Fill additional water bottle for temperature control only. 
 D-9 

6. Record water sample collection on log sheet. 

7. Immediately place samples in small cooler with frozen ice pack (mandatory) 

for transport to the Food Quality Lab, 3375 Koapaka Street, Suite 314. If 

transport is not immediate, place samples in refrigerator. Samples must be 

received no later than 1530 on day of sampling (Monday - Thursday only). Sign 

initials in log book at Food Quality Lab and pick up a sterile sample bottle with 

label and temperature control bottle. Store sample bottles in cooler and ice pack 

in freezer until next sampling. 

8. Results are FAXED within 2-3 days and mailed within a week. These counts 

should not exceed 1000 MF/100 ml. If fecal coliform counts exceed 1000 

MF/100 ml, results are reported to Robert Dollar by phone; sampling must be 

repeated within 24 hours. Promptly notify Dr. Bob Braun if counts are above 

1000 MF/100 ml. 

9. Enter date, time, coliform count, and any pertinent comments in Quattro Pro 

file H:\REHAB\COLIFORM.WQ1. 

DIRECTIONS TO FOOD QUALITY LAB 

The Food Quality Lab is located at 3375 Koapaka Street, Suite 314. On 

Nimitz highway, turn left on Paea Street and drive past the shopping center 

(Airport Mini Mart). Turn right on Ualena Street. Lonestar should be on your 

right and a ramp should be on your left. Drive up the ramp to get to the Food 

Quality Lab. 

VI. VETERINARY CARE/EMERGENCY CARE

1. In case of an emergency or suspected illness, refer to the phone list and call Dr. 

Bob Braun immediately to discuss symptoms or circumstances of emergency or 

illness. If Dr. Braun does not return the call within 5-10 minutes, contact Rob 

Dollar. 

2. A veterinarian or trained NMFS staff member will perform any needed 

biomedical sampling. Collect one lavender top tube (LTT; for CBC) and two 

serum separator tubes (SST; for serum chemistry and serum banking). Fill LTT 

halfway and immediately invert to mix additive with blood (insure blood is not 

clotted). Do not invert SST's. Let SST tubes sit at room temperature for at least 

15-30 minutes (but not more than 1 hour); then spin tubes in centrifuge for 10 

minutes. Draw off serum using pasteur pipet into 2 portions (at least 2 ml to send 

to CVD and remaining sample to freeze in cryovials for monk seal serum bank). 

Label tubes with animal ID, date, NMFS, and sample type. 

3. Medical supplies for blood sampling, fluid and antibiotic administration, crash 

kit, and emergency drugs are in the monk seal lab at KRF. 
 D-10 

4. All monk seal medications and additional medical supplies are kept at the 

monk seal lab, KRF. 

5. CVD forms, current files for rehab seals, and other rehab-related files are kept 

in the monk seal trailer at KRF. 
 D-11 

OVERVIEW OF ACTIVITIES 

1. Conduct initial inspection of seals and enclosure. 

 

2. Weigh out fish and thaw. 

3. Turn on water hose outside of enclosure before feeding. Enter enclosure only 

twice and minimize potential contamination. 

4. USE NOLVASAN FOOTBATH BEFORE AND AFTER ENTERING 

ENCLOSURE. 

5. WASH HANDS PRIOR TO AND AFTER FEEDING. 

6. KEEP DOOR TO ENCLOSURE SECURELY CLOSED AT ALL TIMES. 

7. Feed seals. Note if each seal appears satiated or is still hungry after feeding, and if 

any unusual behavior or clinical symptoms arise. 

8. Clean deck once a day. 

 

9. Wash buckets and other utensils and sanitize sink area. Throw accumulating 

scales in designated garbage bag in freezer. Properly dispose of any uneaten or 

damaged fish. 

10. Succinctly record appropriate information on feeding forms. 

11. Mop floor of fish house. 

12. Check seals one last time before leaving. Ensure gate to enclosure is locked, 

water hose is turned off, and orange drain valve is at appropriate setting. If water 

to tank was turned off/on, recheck main source. NEVER LEAVE IN A HURRY; 

always double check that all tasks are completed. 

13. Return key to seal enclosure to cage and securely lock cage immediately after 

initial unlocking to insure keys are not misplaced. If no one else is on premises, 

make sure all doors and gates are securely locked. 

14. If any behavior or other circumstance arises during feeding that is unusual in 

the recent history of any seal (e.g., lethargy, inappetence, wounds), immediately 

contact Dr. Bob Braun. 
 D-12 
 

APPENDIX E-1: 

Additional Samples Requested During the Laysan 2000 Abortion Investigation
 
 E-1 

 

Abortion Investigation, Laysan 2000 

Master Sample List (Including UME-specific Requests) 

Histology: Samples, preservation, amount and container will be as 

described in the latest Terry Spraker Field Necropsy Protocol. 

Addition: Per the request of Terry Spraker two samples 2.5 cm x 5 cm x 

5 cm of lung will be put into formalin. 

Hematology: Samples, preservation, amount, and container will be as 

described in the HMS Sample Protocol and performed as in the past 

epidemiology tier one samples (see Hawaiian monk seal necropsy and 

specimen collection protocols). 

Cytology: Two impression smears each from each lesion will be taken 

(one air dried and one fixed in 90% ehtono). 

PCR: 

Live sampling: swab natural orifices (genital, ocular, rectal, 

oral, and nasal) (sterile swab in cryovial, frozen -20° C) 

Necropsy Sampling (2 to 3 sets of tissues): Lymph nodes (mediastinal, 

mesenteric, and tonsils), liver, lung, spleen, and kidney (1 cm3, in 

whirlpak bag, frozen -20° C) 

Biotoxin: 

Live Sampling: feces (plastic bag 1-100 g, frozen -20° C) 

Necropsy Samples (1 to 3 sets of samples as possible): Liver, heart, 

skeletal muscle, feces, stomach contents (1-100 g in whirlpak bag, 

frozen -20° C); Whole blood and urine (1-100 ml in Teflon container) 

Other Toxins: Live Sampling: Whole Blood (5 ml in Teflon or 

scintillation vials, frozen -20° C), blubber sample (6 mm biopsy punch, 

amber tox vial or teflon jar, frozen -20° C) 

 

Necropsy Samples (see necropsy protocol) 

If Euthanasia: Skeletal muscle, liver (100 g, foil, frozen -20° C) 

Fatty Acid: Live sampling: Blubber biopsy (6 mm biopsy punch, 5 ml 

cryovial, frozen liquid nitrogen) 

Bact/Virology: 

Live Sampling: Swabs (rectal, genital, ocular, oral, and 

nasal) (in cryovials frozen in liquid nitrogen), Salmonella rectal 

swab, aerobic/anaerobic culturette (rectal). One nasal swab (sterile 

cryovial, in transport media, frozen in liquid nitrogen) 
 E-2 

 

Parasitology: Feces in PVA and GI preserved per Field Necropsy Protocol 

and Specimen Collection Protocols 

Additional: Scats, Spews-Normal protocol. If fresh, frozen in plastic 

bags (frozen -20° C) 
 E-3 

 

APPENDIX E-2 

Specimen Collection Protocols for Specific Biotoxins 

Reprinted with permission from: Rowles, T.K., F.M. Van Dolah, and A.A. Hohn. 

2001. Gross necropsy and specimen collection protocols. Pp. 449 - 470 In: 

CRC Handbook of marine mammal medicine (Dierauf, L.A. and F.M.D. Gulland, 

eds.). 2nd edition. CRC Press, Boca Raton. Copyright CRC Press, Boca 

Raton, Florida.
 E-4 

 
 E-5 
 E-6 
 E-7 

 

APPENDIX E-3 

Specimen Collection Protocols for Chemical Pollutants 

Reprinted with permission from: Rowles, T.K., F.M. Van Dolah, and A.A. Hohn. 

2001. Gross necropsy and specimen collection protocols. Pp. 449 - 470 In: 

CRC Handbook of marine mammal medicine (Dierauf, L.A. and F.M.D. Gulland, 

eds.). 2nd edition. CRC Press, Boca Raton. Copyright CRC Press, Boca Raton, 

Florida 
 E-8 

 
 E-9 

 

 
 E-10 
APPENDIX F-1: 

Epidemiology Sampling Supply List

 F-1 

Epidemiology sampling supply list 

LABORATORY SUPPLIES 

glass slides 

cover slips 

Slide holders 

Unopette WBC System (100) 

Capillary pipettes for Unopette 

critoseal (tube sealer) 

Hemacytometer Chamber 

Hemacytometer cover glass 

Lab counter 

Kim wipes 

Quickcheck test strips BUN 25/bottle 

Glucose test strips/50 strips/vial 

Accucheck Advantage Glucometer 

glucometer control solution 

Disposable transfer pipettes 

Non-heparinized HCT tubes 

latex gloves 

wooden applicator sticks 

LABORATORY EQUIPMENT 

Clinical Refractometer 

Microscope 

Spare microscope bulbs 

lens paper 

lens cleaner 

Centrifuge 

Microhematocrit centrifuge 

light for reading HCT 

SWABBING/FECAL COLLECTION 

Virology transport media in 1.8 ml cryovials 

Virology swabs 

Bacteriology swabs -Culturette 

C&S bacteriology media (for Salmonella) 

PVA fixative solution 32 oz 

vials 

fecal loops 

KYJel 

Digital thermometer (rectal) 

BIOPSY SUPPLIES 

biopsy punches 6 mm 

disposable scalpel blades #11 

forceps 

scissors 

xylocaine 2% 50 ml 

disposable sterile glove 

scintillation vials 

amber vials 

cryovials, 5.0 ml
 F-2 

CRYOVIALS 

1.0 ml 

1.8 ml 

5.0 ml 

nalgene marking pens 

SYRINGES 

35 cc slip tip 

60 cc 

12 cc slip tip 

6 cc 

IV extension sets 

NEEDLES 

18 gx3.5" 

18 gx 1.5" 

22gx1.5 

VACUTAINER TUBES 

SST 9.5 ml 

LTT 3 ml 

GNTT 5 ml 

GNTT 9 ml 

DISINFECTANTS, SCRUBBING, ETC 

isopropyl 

liter Betadine solution 

Nolvasan 1 gallon 

scrub brush 

gauze 

distilled water 1 gallon 

bleach 

paper towels 

bottles for scrubbing 

soap for handwashing 

HANDLING SUPPLIES 

tape measure for length and girth 

coveralls 

masks 

hoop net 

boots 

cotton gloves 

knee pads 

calipers 

FIELD FORMS 

necropsy 

clinical exam 

restraint 

lab sheets 

specimen collection log 

controlled drug log 

specimen labels 
 F-3 

MISCELLLANEOUS 

cooler 

Nitrogen Dewar 

batteries 

pens 

pencils 

sharps containers 

Biohazard bags 

latex gloves 

Photographic camera w/lens 

Film - 400 ASA slides/paper 
 F-4 
 F-5 

APPENDIX F-2: 

Veterinary Medical Kit
 F-6 
 F-7 

VETERINARY MEDICAL KIT 

Ambubag (resuscitation) bag 2.0 L bag 

stylet for 10.5" ID endotracheal tube 

Lactated Ringer's 

0.9% Sodium chloride injection 1 L 

Littman Master cardiology stethoscope 24" 

Dexamethasone (4mg/ml; 100 cc) 

Atropine 1/120 per ml (100 ml) 

Diazepam 5 mg/ml vials 

Epinephrine 1:1000 20 cc 

Soludeltacortef 500 mg; 20 cc 

Dopram IV (20mg/ml; 100 

8.4% sodium bicarbonate 

50% dextrose 

Endotracheal tube 14 mm 

Endotracheal tube 12 mm 

Endotracheal tube 10 mm 

IV drip set 

hemostats 

scalpel handle 

sterile scalpel blades 

needle holders 

absorbable suture 

18 gx1.5" catheters 

18 gx 5" catheters 

3 cc syringes 

20 cc syringes 

rope

APPENDIX G-1: 

Monk Seal Specimen Collection Summary Form

 G-1 

 MONK SEAL SPECIMEN COLLECTION SUMMARY: ____________________ ___________________ 

Island Year 

Specimen Preservation Method Diagnostics Speci

Num Sub 

Num 

Type Sub 

Type 

Seal 

ID 

Temp 

ID 

Size Sex 

Survival 

Factor 

Num 

Collect 

Date 

Islet Collection Transport Arrival Transport 

Means 

Date 

 Sent

Date 

Arrived

Lab Date 

Sent 

Eval. Result Date 

Tested 

Facility Loc

SPECIMEN TYPE: SUB-TYPE: PRESERVATION METHOD: BB = blubber biopsy PL = the ‘pluck’ WC = whole carcass BT = blue top tube RT = red top tube AA = absolute alcohol (98%) BC = blood cells PS = plasma WH = whole heart DN = DNA anal. SP = spleen AF = AFA 50ml 95% ethyl alcohol BL = blood RS = rectal swab ON = other necropsy GI = GI tract SS = serum sep. tube 5ml acetic acid, 45ml of 10% formalinBU = buffy coat RT = reprod. tract OS = other swab GN = green top tube VM = viro. medium AG = 95ml 70% ethyl alcohol, 5ml gly BS = blood smear SC = scats OT = other GT = grey top tube PROJECT: AL = 70% ethanol 

ES = eye swab SE = serum NO = none (sample discarded) KI = kidney UM = UME CB = chloroform, BHT mix 

FL = fecal loop SK = skeletal SUB-TYPE: LI = liver EP = epidemiology DM = DMSO GI = gastrointest. Tract SP = spew AD = adrenal LT = lavendar top PA = pop. assess. DR = dry NMO = molt ST = stomach contents BC = blubber chunk (necr.) LU = lung PS = pup surv FR = frozen NS = nasal swab TP = tissue plug BM = bact. medium NP = not processed ST = sat. tag. FM = 10% buffered formalin NT = necropsy tissue VS = vaginal swab BP = blubber plug (necr.) OL = otolith CC = crit. cam PC = placenta WB = whole blood BR = brain PA = parasite OT = other 
 G-2 

MONK SEAL SPECIMEN COLLECTION SUMMARY: __________________ _____________ 

 Island Year 

Specimen  no. Notes 
 G-3 

 

APPENDIX G-2: 

Hawaiian Monk Seal Tagging & Chemical Immobilization Form
G-4
G-5
G-6
G-7

 

APPENDIX G-3: 

Hawaiian Monk Seal Clinical Examination Form
G-8
G-9
G-10
G-11

 

APPENDIX G-4: 

Hawaiian Monk Seal Field Hematology Report 
G-12
G-13
G-14
G-15

APPENDIX G-5: 

NMFS Human Interaction Forms 
G-16
G-17
G-18
G-19

 

APPENDIX G-6: 

Marine Forensics Chain of Custody Form (NMFS, SEFSC) 
G-20
G-21
 
 

 

APPENDIX H-1: 

Federal, State and Local Authorities 
 
 H-1 

Name Title Office Address Phone E-mJeffrey Polovina 

Acting Science Center Director, PIFSC PIFSC 

2570 Dole St, Honolulu, HI 96822 808-983-5390 Jeff

George Antonelis 

Chief, Protected Species Division PIFSC 

2570 Dole St, Honolulu, HI 96822 

808-983-5710 808-226-6544 (c) Bud

Jason Baker 

Leader, Hawaiian Monk Seal Population Assessment PIFSC 

2570 Dole St, Honolulu, HI 96822 808-983-5711 Jaso

Robert Dollar 

Manager, Kewalo Research Facility KRF 

1125B Ala Moana Boulevard, Honolulu, HI 96814 

808-592-8302 808-226-0525 (c) 808-660-4893 (p) Rob

Samuel Pooley 

Acting Regional Director, PIRO PIRO 

1601 Kapiolani Blvd, Honolulu, HI 96814 

808-973-2935 x222 Sam

Margaret Akamine 

Resource Management Specialist PIRO 

1601 Kapiolani Blvd, Honolulu, HI 96814 

808-973-2935 x210 808-721-5340 (c) Ma

Bradley Ryon 

Protected Resources Management Liaison PIRO 

1601 Kapiolani Blvd, Honolulu, HI 96814 

808-973-2935 x258 808-721-5341 (c) Bra

Allen Tom Sanctuary Manager 

Hawaiian Islands Humpback Whale NMS 

Headquarters Office – Maui, 726 South Kihei Rd, Kihei, HI 96753 800-831-4888 Alle

Robert Smith Reserve Coordinator 

NW Hawaiian Islands Coral Reef Ecosystem Reserve Office 

308 Kamehameha Ave, Suite 203 Hilo, HI 96720 808-933-8181 Rob

Aulani Wilhelm 

Assistant Reserve Coordinator 

NW Hawaiian Islands Coral Reef Ecosystem Reserve Office 

6700 Kalanianaole Hwy, #215, Honolulu, HI 96825 808-397-2657 Aul

Laurie Allen Acting Director 

Office of Protected Resources 

1315 East-West Highway, Silver Spring, MD 20910 (301) 713-2319 Lau

Teri Rowles 

Marine Mammal Health and Stranding Program 

NMFS Office of Protected Resources 

1315 East-West Highway, Silver Spring, MD 20910 

(301) 713-2322 x178 Ter

Janet Whaley 

Marine Mammal Health and Stranding Program 

NMFS Office of Protected Resources 

1315 East-West Hwy, Silver Spring MD20910 

(301) 713-2322 x170 Jan
 H-2 

Name Title Office Address Phone E-mGordon Helm Media Contact 

National Marine Fisheries Service 

 (301) 713-2370 

x140 Gor

Wende Goo 

Management Analyst/ Community Outreach Coordinator PIFSC 

2570 Dole St, Honolulu, HI 96822 808-983-5303 We

Elton Hout 

Office of Ocean and Coastal Resource Management National Ocean Service 

1305 East-West Hwy, Silver Spring, MD 20910 

(301) 713-3155 x200 Elto

Sylvia Galloway 

Center Director, National Centers for Coast Ocean Science 

NOAA National Ocean Service 

219 Fort Johnson Rd, Charleston, SC 29412 843-762-8525 Syl

Office of Response and Restoration 

NOAA National Ocean Service 

1305 East-West Hwy, Silver Spring, MD 20910 (301) 713-2989 

Tim Bodeen Refuge Manager 

USFWS Midway Atoll NWR 

USFWS Midway Atoll P.O. Box 50167 Honolulu, HI 96850 

808-674-8237 x102 tim 

 USFWS Hawaii and 

Pacific Islands NWR Complex 

300 Ala Moana Blvd, Box 50167, Honolulu, HI 96820 808-541-1201 

Capt. Timothy V. Skuby Commanding Officer 

U.S. Coast Guard Marine Safety Office 

433 Ala Moana Blvd, Honolulu, HI 96813 808-522-8251 tsku

Jeffrey Walters 

State Sanctuary Comanager State of HI, Dept of Land & Natural Res 1151 Punchbowl St, Rm 330, Hon, HI 96813 808-587-0106 Jeff

hi.u

Paul Conry 

Wildlife Program Manager 

State of Hawaii, DLNR, Division of Forestry and Wildlife 

1151 Punchbowl St, Rm 325, Honolulu, HI 96813 808-587-4176 Pau

Dave Smith Wildlife Biologist 

State of Hawaii, DLNR, Division of Forestry and Wildlife 

2135 Makiki Heights Dr, Honolulu, HI 96822 808-973-9786 viki

William S. Devick Administrator 

State of Hawaii, DLNR Div of Aquatic Res 

1151 Punchbowl St, Rm 330, Hon, HI 96813 808-587-0100 will

Information cur
 H-3 

APPENDIX H-2: 

Potential Response Team Members
 H-4 
 H-5 

VETERINARIANS 

Name Address Phone Pager/Cell E-maDr. Bob Braun 44-299 Kaneohe Bay Drive, Kaneohe, HI 96744 808-254-8181 C 808-783-6565 

rbraun

Rober

Dr. Les Dalton 

SeaWorld of Texas, 10500 SeaWorld Dr., San Antonio, TX 78251 210-523-3278 P 210-746-0232 Les.D

Dr. Chris Dold 

The Marine Mammal Center, 1065 Fort Cronkite, Sausalito, CA 94965 415-289-8609 P 415-258-3539 doldc

Dr. Frances Gulland 

The Marine Mammal Center, 1065 Fort Cronkite, Sausalito, CA 94965 415-289-7370 P 415-257-9613 gullan

Dr. Marty Haulena 

The Marine Mammal Center, 1065 Fort Cronkite, Sausalito, CA 94965 415-289-7370 P 415-258-3983 haule

Dr. Gregg Levine 267 S. Kalaheo Avenue, Kailua, HI 96734 808-261-7031 C 808-358-5311 glevinDr. John Reif 

Colorado State University, Dept. of Environ. Health, Fort Collins, CO 80523 970-491-6074 john.r

Dr. Terry Spraker 

Colorado State University, College of Veterinary Medicine, Fort Collins, CO 80523 970-297-4155 tsprak

Dr. Pam Yochem 

Hubbs-SeaWorld Research Institute, 2595 Ingraham St., San Diego, CA 92109 619-226-3874 P 619-242-3551 pyoch

Information curren
 H-6 

 

 

 

 
 H-7 

 

 

 

APPENDIX H-3: 

Diagnostic Laboratories
 H-8 

 

 
 H-9 

 

 

Information current, November 2003

 

 Laboratory Analysis Phone Address 

 

IDEXX Serum biochemistry 800-444-4210 

2825 KOVR Drive, West 

Sacramento, CA 95619 

Dr. Jeff Stott 

U.C. Davis, Laboratory for 

Marine Mammal Immunology Virology, immunology 530-752-3349 

Dept. of Pathology, Microbiology 

and Immunology, 

School of Veterinary Medicine 

2054 Haring Hall, 1Shields Ave. 

Davis, CA 95616 

Spencer Jang 

U.C. Davis, Veterinary 

Medical Teaching Hospital Bacteriology 530-752-1168 

VMTH 

Microbiology Dept./Lab 1011 

1 Garrod Drive 

Davis, CA 95616 

Dr. Dori 

Borjesson 

University of Minnesota, 

Dept. of Diagnostic Medicine WBC differentials 612-624-7400 

Veterinary Diagnostic Laboratory 

1333 Gortner Avenue 

St. Paul, MN 55108 

Dr. Klaus Nielson 

Animal Disease Research 

Institute/Canadian Food 

Inspection Agency Infectious diseases 613-228-6698 

Canadian Food Inspection Agency 

3851 Fallowfield Road 

Nepean, Ontario, Canada K2H8P9 

Dr. Ole Nielson 

Department of Fisheries and 

Oceans Canada Infectious diseases 204-983-5126 

501 University Crescent, 

Winnipeg, Manitoba, Canada R3T 

2N6 

Dr. Beverly 

Schmitt 

National Veterinary Services 

Laboratory Bacteriology 515-663-7266 1800 Dayton Rd, Ames, IA 50010 

Dr. Jeremiah 

Saliki 

Oklahoma Animal Disease 

Diagnostic Lab Virology 405-744-8809 

Farm Road and Ridge 

Stillwater, OK 74078 

Dr. S.K. Hietala 

CA Animal Health and Food 

Safety Laboratory Bacteriology 530-752-7578 

Thurman Lab West Health Services 

University of CA, Davis 

Davis, CA 95616 

Dr. Tod 

Leighfield 

NMFS Marine Biotoxins 

Program Toxicology 843-762-8500 

219 Fort Johnson Rd, Charleston, SC 

29412 

Dr. Gina Ylitalo 

Environmental Conservation 

Division, NFSC Toxicology 206-860-3325 

2725 Montlake Blvd East, Seattle, 

WA 98112 

Dr. Sara Iverson Department of Biology Fatty acid analysis 902-494-2566 

Dalhousie University, Halifax, Nova 

Scotia, B3H 4J1, Canada 

Dr. Tom 

Lipscomb 

Department of Veterinary 

Pathology, AFIP Histopathology 202-782-2600 

6825 16th St. NW, Washington, D.C. 

20306 

Dr. Terry Spraker Colorado State University Histopathology 970-419-4155 

College of Veterinary Medicine, Fort 

Collins, CO 80523 

Dr. Michael Kliks CTS Foundation Parasitology 808-988-7203 3081 G. Paty Dr, Honolulu, HI 96822

Dr. Murray Dailey The Marine Mammal Center Parasitology 415-289-7346 

1065 Fort Cronkite, Sausalito, CA 

94965 
 H-10 

 

 
 H-11 

 

 

 

 

APPENDIX H-4: 

Cooperating Experts
 H-12 

 

 
 H-13 

 

 

Cooperating Experts 

Name Address Phone E-mail 

A. Alonso Aguirre 

Wildlife Trust, 61 

Route 9W, Palisades, 

NY 10964-8000 845-365-8368 aguirre@wpti.org

Murray Dailey 

The Marine Mammal 

Center 

Marin Headlands 

1065 Fort Cronkhite 

Sausalito, CA 94965 

415-289-7346 or 

415-289-7360 daileym@tmmc.org

William G. Gilmartin 

Hawaii Wildlife Fund, 

P.0. Box 70, Volcano, 

HI 96785 808-985-7041 bill-gilmartin@hawaii.rr.com

Yoshitsugi Hokama 

University of Hawaii 

at Manoa, Biomed D105 

Honolulu, HI 96822 808-956-5464 hokamay@jabsom.biomed.hawaii.edu

Michael Kliks 

CTS Foundation 

3081 G. Paty Drive 

Honolulu, HI 96822 808-988-7203 mmkliks@hawaii.rr.com

Yuanan Lu 

Retrovirology 

Research Laboratory, 

Pacific Biomedical 

Research Center 

University of Hawaii 

at Manoa 

Leahi Hospital 

3675 Kilauea Avenue 

Honolulu, HI 96816 808-732-7702 ylu@pbrc.hawaii.edu

James McBain 

SeaWorld of 

California, 500 

SeaWorld Dr, San 

Diego, CA 92109 619-226-3833 james.mcbain@anheuser-busch.com

Ole Nielsen 

Central and Arctic 

Region Fishereies and 

Oceans Canada 501 

University Crescent 

Winnipeg, Manitoba, 

Canada R3T 2N6 204-983-5126 

Stephen Rafferty 

Animal Health Center 

1767 Angus Campbell 

Road 

Abbotsford, BC 

Canada 

V3G 2M3 604-556-3003 

Timothy Ragen 

Marine Mammal 

Commission 

4340 East-West 

Highway 

Bethesda, MD 20814 301-504-0087 tragen@mmc.gov

 Information current, November 2003 
 H-14 

 

 

 
 

 

APPENDIX I-1: 

Flowchart and Timing of Response to MMUME in the United States 

 
 
 I-1 

 

 I-3 

 

 

APPENDIX I-2: 

Flow Diagram for Suspected Marine Biotoxin Incidents 

(NOAA Marine Biotoxins Program, 

http://www.chbr.noaa.gov/CoastalResearch/Pictures/AnalyticalResponseTeam/FlowDiag

ram.gif) 

 I-5 

 

Suspected 

Algal Bloom 

Collect Water Samples 

for Algal Identification 

Perform Mouse 

Bioassay 

Perform Toxin Class 

Specific Analysis 

Is there evidence of an 

algal bloom? Go to 1. 

Unusual Annual 

Mortalities 

Is there evidence to support 

Phycotoxin Involvement Based 

on Symptoms? 

Collect stomach contents, 

serum, prey species and 

sentinel species 

Is there evidence of an 

algal bloom? 

Collect stomach liver, 

kidney, prey species, 

and sentinel species 

NO 

NO 

YES

YES

Producer

Not Toxic 

Known

Perform Battery of Assays to 

Determine Mode of Action 

STOP 

Live 

Stranding 

Known 

Toxin 

+

_

Address 

Other 

Causes 

Address 

Other 

Causes 

Non-toxic 

bloom 

Go to 1. 

Flowchart Diagram for Suspected Marine Biotoxin Incidents 
 I-6 

 
I-7

APPENDIX I-3: 

Flow Chart for Response to a Marine Mammal Mortality Event 

with Known vs. Unknown Cause 

(Wilkinson, D.M. 1996. National Contingency Plan for Response to Unusual Marine 

Mammal Mortality Events. NOAA Tech. Memo NMFS-OPR-9, 118p. Table 1, 

between pp. 4 and 5). 

 I-9 

 
 I-10 

 
 

 

APPENDIX J-1: 

Memorandum of Understanding with Participants in Unusual Mortality Event 

Response

(Geraci, J.R. and V.J. Lounsbury. 1997. DRAFT Contingency Plan for Manatee DieOffs. Prepared for Florida Departement of Environmental Protection. Appendix E.) 
 
 J-1 

 

Example 
 J-2 

 
 J-3 

 

 

APPENDIX J-2: 

Memorandum of Understanding with Laboratories/Individuals Performing 

Tests/Analyses 

Related to Unusual Mortality Event Response Investigation

(Geraci, J.R. and V.J. Lounsbury. 1997. DRAFT Contingency Plan for Manatee DieOffs. Prepared for Florida Departement of Environmental Protection. Appendix E.) 

 
 J-4 

 
 J-5 

 

Example 
 J-6 

 
 

APPENDIX K-1: 

Hawaii Area Oil Spill Contingency Plan 

Excerpts from 28 August 2001 Update of the Hawaii Area Contingency Plan

 (www.uscg.mil/d14/units/msohono/HACP1/index.htm)
 
 K-1 
 K-2 
 K-3 
 K-4 
 K-5 

 

APPENDIX K-2: 

Hawaii Area Oil Spill Contingency Plan 

Excerpts from Oil Spill Field Operations Guide

( www.uscg.mil/d14/units/msohono/ics/fog/index.htm) 
 K-6 
 K-7 
 K-8 
 K-9 
 K-10 
 K-11 
 K-12 
 

Availability of NOAA Technical Memorandum NMFS

Copies of this and other documents in the NOAA Technical Memorandum NMFS series 

issued by the Pacific Islands Fisheries Science Center are available online at the PIFSC 

Web site http://www.pifsc.noaa.gov in PDF format. In addition, this series and a wide 

range of other NOAA documents are available in various formats from the National 

Technical Information Service, 5285 Port Royal Road, Springfield, VA 22161, U.S.A. 

[Tel: (703)-605-6000; URL: http://www.ntis.gov]. A fee may be charged. 

Recent issues of NOAA Technical Memorandum NMFS-PIFSC are listed below: 

NOAA-TM-NMFS-PIFSC-1 The Hawaiian monk seal in the Northwestern Hawaiian 

 Islands, 2001. 

 T. C. JOHANOS and J. D. BAKER (comps. and eds.) 

 (April 2004)