Authorizations and Permits for Protected Species (APPS)

I. PROJECT PURPOSE 
In June 2000, the National Marine Fisheries Service (NMFS) and the Aleut Community of St. Paul Island Tribal Government (ACSPI) signed the original co-management agreement for co-management of northern fur seals (NFS) and Steller sea lions (SSL) on St. Paul, Walrus and Otter Islands and Sea Lion Rock and finalized an amended agreement that includes co-management of harbor seals in January 2020. The ACSPI and NMFS identified the following co-management responsibilities regarding the research of NFS, SSL, and harbor seals: long-term data collection programs, sampling programs, population abundance and status, habitat use and seasonal movements, sources of natural and human-caused mortality, and disentanglement programs. The activities described herein are in fulfillment of our co-management responsibilities and will be undertaken by the ACSPI’s Ecosystem Conservation Office (ECO) department in collaboration with NMFS.
Project 1. Marine Mammal Population Monitoring/Assessment
The objectives of the marine mammal population monitoring/assessment study are to provide the Marine Mammal Laboratory (MML) with small unmanned aircraft systems (sUAS) imagery to assess abundance of harbor seals and SSL and information on habitat use and seasonal movements of overwintering NFS and branded SSL, respectively. We will use aircraft sUAS to survey and photograph harbor seals, SSL, and NFS in the Pribilof Islands to obtain images for counts and/or photogrammetry. The ECO will coordinate sUAS flights with MML to avoid duplication of MML’s current research on harbor seals, SSL, and NFS. The ECO is in a unique position to access Otter and Walrus Islands and provide MML with regular aerial images and count data to estimate pinniped population sizes and trends and consistent brand re-sight data to estimate SSL survival and immigration/emigration rates.
ECO will conduct sUAS flights annually to obtain images of harbor seal pups and non-pups on Otter and St. Paul Island and opportunistically on St. George Island, and SSL pups and non-pups on Walrus Island rookery and opportunistically on Otter Island. ECO will prioritize Otter Island for harbor seals and Walrus Island rookery for SSL. Aerial surveys of overwintering NFS non-pups will be conducted during winter and spring months (i.e., December-May) on St. Paul and Sea Lion Rock and at other locations as deemed viable. Aerial surveys of branded SSL will be conducted throughout the year on St. Paul and Sea Lion Rock. Additionally, ECO will conduct sUAS surveys of NFS haulouts and rookeries on St. Paul Island to monitor marine debris accumulation rates to evaluate habitat conditions for NFS. These data are critical for future marine debris removal and monitoring efforts, as they will help to identify marine mammal habitats that are most vulnerable to marine debris accumulation. By identifying these regions, ECO can adjust future removal efforts to focus on the most vulnerable regions and apply increased removal efforts to these regions.
Harbor seals
The Pribilof Islands stock of harbor seals is one of 12 stocks in Alaska; surveys of the isolated and at-risk Pribilof Islands stock historically have been minimal and opportunistic. All other Alaska harbor seal stocks are routinely surveyed and photographed from a manned aircraft using oblique and vertical photography. Due to limited funds and the remote geographic location of the Pribilof Islands, it’s rarely feasible for MML to conduct aerial surveys of this stock. Counts of harbor seals in the Pribilof Islands ranged from 250 to 1,224 in the 1970s and between 119 and 232 in the 1980s and 1990s. Prior to July 2010, the most recent count was in 1995 when a total of 202 seals were counted. In July 2010, approximately 185 adults and 27 pups were observed on Otter Island plus approximately 20 on all the other islands combined for a total of 232 harbor seals. Maximum seal counts (all ages) are nearly identical to the 1995 counts (212 vs. 202), but 2010 pup numbers were slightly less (27 vs. 42). In 2018, ECO conducted a successful pilot project with MML’s Polar Ecosystems Program and Duke University to evaluate the use of sUAS to collect imagery for harbor seal stock assessment. This effort provided MML with the first ever comprehensive survey of Pribilof Island harbor seals. The total average count from the recent 2018 and 2019 sUAS surveys is 240 seals; using an approximate correction factor for seals likely in the water, the current abundance estimate for the Pribilof Island harbor seal stock is 540 seals. This project activity has opened up opportunities to expand sUAS operations in the Pribilof Islands using local capacity for enhancing marine mammal programs. Harbor seal data are incorporated into MML’s broader population monitoring of harbor seals in Alaska and have facilitated exploration of other potential project partnerships between ECO and MML regarding marine mammal research.
Steller sea lions
Walrus Island is located 10 miles northeast of St. Paul Island; it is not part of the usual MML aerial survey route or trend sites but is occasionally included in the western distinct population segment (DPS) survey flight plan for SSL. Over the last 50 years, SSL pup production on Walrus Island has declined by over 90%, from 2,866 pups born in 1960; pup production for the central Aleutian Islands declining 1.33% annually from 2002-2017 (Sweeney et al. 2017). MML conducted SSL counts on Walrus Island in 2010 and 2013. In 2010 and 2013, only 14 and 28 pups were counted, respectively, and reflect a 7.34% annual rate of decline in pups since 2000 (NMML 2014). MML was not able to survey Walrus Island in 2011 due to persistent fog and bad weather and did not survey in 2012. In addition to the summer breeding rookery on Walrus Island, SSL use haulout sites on St. Paul, St. George, and Otter Islands throughout the year. The numbers, age-classes, and genders of SSL at these sites vary seasonally. SSL pups usually remain within 500 km of their natal site during their first year and lactating female SSL often move with their pups from the natal rookery to winter haulout sites near more productive foraging grounds (Raum-Suryan et al. 2002). Given the geographical isolation of the Pribilof Archipelago, the winter haulout sites on St. Paul and St. George are very likely to be important for the Pribilof breeding population during winter, as well as for SSL foraging from other areas. Local observations of branded SSL have demonstrated animal movement between the Pribilofs and southeast Alaska, the Gulf of Alaska, Aleutian Islands, and Russia. ECO and local subsistence hunters are able to provide MML with consistent brand re-sight information throughout the year on St. Paul Island and Sea Lion Rock to estimate survival and immigration/ emigration rates of SSL.
Northern fur seals
Adult NFS spend about 80% of their time at sea. During the winter and spring, they are pelagic, occupying the North Pacific Ocean as well as the Bering and Okhotsk Seas. Territorial adult male NFS leave their breeding colonies in August and are thought to spend most of their time in the Bering Sea and North Pacific Ocean along the Aleutian Islands. In recent years ECO has noticed an increase in the number of male NFS offshore of St. Paul Island and hauling out on Sea Lion Rock during the winter months. In order to document potential changes in the abundance and locations of overwintering NFS ECO began surveying (using binoculars) in 2016 the number of NFS hauling out on land during the winter and spring months. In February 2019, ECO counted between 800-1,000 NFS hauled out on Sea Lion Rock (ECO, unpub. data). The implementation of sUAS for marine debris surveys will increase our understanding of accumulation rates of various types of marine debris, including entangling debris, on rookeries and haulouts in the Pribilof Islands. Utilizing sUAS for these surveys allows us to cover much more ground and more accurately track marine debris while minimizing marine mammal disturbance.
Numbers of individuals incidentally disturbed during each population monitoring/assessment activity are determined using abundance estimates, the proportions of animals in different age/sex categories present (on land) during the surveys and estimates of the proportion of animals present that are disturbed by each activity based on previous experience of ECO staff MML researchers, and other previously permitted research scientists. These numbers account for the abundance and trends at the specific rookeries where, and the seasonal behavior/attendance patterns when, the activities are conducted and are estimated as maximum potential disturbance.
Project 2. Northern Fur Seal Disturbance using VHF
The objective of the NFS VHF study is to investigate adult female NFS response to subsistence and research activities at Northeast Point (NEP) on St. Paul Island. ECO and MML will work together as co-management partners to track and record the location, timing, and duration of subsistence and research activities directed at NFS to better understand the consequences of human presence within NFS habitat. This study complements MML’s ongoing and long-term VHF emigration work (assessment of demographic parameters) on rookeries on both islands to detect emigrants that might have left the rookeries where they were originally tagged.
Preliminary estimates from ongoing studies at the Pribilof Islands indicate that both juvenile and adult survival rates are lower than historical estimates, and that either could explain the observed declines on their own. These results support the implementation of additional field (emigration) studies to determine the impacts of emigration on estimates of survival. Emigration from focal study rookeries will be estimated by deploying VHF tags and monitoring with receivers will occur at other rookeries to identify emigrants.
Pulse-coded very high frequency (VHF) transmitters have been used to detect variation in attendance behaviors of adult female NFS at multiple sites in the Pribilof Islands in recent years (e.g., Kunisch 2011; NMFS, unpub. data). VHF flipper tags have allowed MML to detect permanent (emigration) and temporary (disturbance) movements away from NFS tagging sites to alternative sites where VHF receiver stations are placed. Programming the transmission cycles of VHF tags enables detection of permanent movements (i.e., emigration) and short-term behavioral responses (i.e., temporary disturbance) of NFS at different breeding areas in the Pribilof Islands. Improved detection and categorization of movements will provide critical insight into the declining NFS population segment on St. Paul Island. The utility of VHF tags is ideal for characterization of short-term behavioral responses and the long-term consequences of those responses of NFS by examining variation in other indices of behavior or health. ECO, MML, and NOAA Fisheries Alaska Region will collaborate to maximize data collection for disturbance from subsistence, research, and other activities; seasonal attendance; and estimation of emigration rates within and between breeding sites.
ECO will use the specific VHF transmitter and receiver programming used previously by MML in a validation study on St. George and St. Paul Islands to maximize data collection for seasonal attendance, and estimation of emigration rates within and between breeding sites on islands. The VHF transmitters will be programmed in advance to transmit at particular rates for their entire deployment. Increasing transmission rates to reliably detect short-term movements to other sites decreases the battery life, so this project will attempt to balance the longest battery life with detection of short- and long-term movements. ECO will track and record the location, timing, and duration of subsistence and research activities directed at NFS. MML will share attendance data from other sites where tags have been deployed previously to help characterize and interpret short-term responses of NFS, as well as account for site specific variation in attendance behavior. ECO will share NEP attendance data for MML to use in estimating emigration rates.
In recent years St. Paul Island has observed rapid and dramatic changes in the distribution and abundance of NFS. NFS were listed as depleted under the Marine Mammal Protection Act in 1988 (53 FR 17888). Historically, a majority (i.e., 75%) of the worldwide population of NFS bred on the Pribilof Islands (St. Paul and St. George Islands; Gentry 1998). In 2018, that estimate dropped to approximately 40.5% of the worldwide population, with a majority of NFS breeding on St. Paul Island (J. Sterling, NMFS, pers. comm.). St. Paul and St. George population segments have been in decline since the 1950s, when experimental female culling of the herd occurred (NMFS 1984). Since 1998, pup production on St. Paul Island has declined by 57.7%, at an annual rate of 4.09% (SE ± 0.34; Towell et al. 2018). From 2016 to 2018 specifically, pup production for St. Paul decreased by approximately 6.1% (Towell et al. 2018). Concurrently, the total number of territorial males associated with females on St. Paul Island has decreased by 3.2% and the total number of idle males (without females) decreased by 11.5% from 2017 to 2018 (Towell et al. 2018).
The sample sizes for tagged animals are based on conversations with MML and NOAA Alaska Region personnel and are determined to be an adequate number of animals to be able to detect local disturbances at NEP while accounting for the limited number of personnel available for tagging activities on island. Numbers of individuals incidentally disturbed during animal captures are determined using encounter rates, estimates of the numbers of animals that are disturbed during capture activities, and the relative proportions of animals in different age/sex categories present (on land) at the time of the captures. Numbers of animals incidentally disturbed during monthly VHF receiver data downloads and maintenance are determined using abundance estimates, the proportions of animals in different age/sex categories present (on land) during data downloads and maintenance and estimates of the proportion of animals present that are disturbed by this activity. These numbers also consider trends in abundance, seasonal behavior and attendance patterns, and the terrain at the specific study location and associated observation points.
Project 3. Northern Fur Seal Habitat Use Studies
The objectives of the habitat use studies are to determine the timing of arrival of adult male NFS in the spring, index counts of adult female and harem bull NFS in the summer, and timing of departure of adult female and pup NFS in the fall. Timing of arrival and departure of NFS are fundamental to the management of regulatory closures (50 CFR 216.81) that preclude general public access to NFS breeding and resting areas from June 01 until October 15 on St. Paul Island. Female index counts have the potential to provide information about population trends that are not currently being employed for population assessments (W. Testa, NMFS, pers. comm.). NMFS owns and administers land on St. Paul for the conservation of NFS. This land is often referred to as the rookery; however, the land actually includes both breeding (locally known as the rookery) and resting (hauling grounds or haulout) habitat occupied during the spring, summer, and fall as well as an adjacent buffer area not typically occupied by breeding or resting NFS. ECO focuses observations on the areas not occupied by NFS or typically occupied by non-breeding NFS when observations can reliably be obtained from these areas. We have attempted to distinguish breeding areas occupied by non-pups from resting areas occupied by non-breeding non-pups. NFS habitat use studies will be conducted annually at least two times per week during the spring (April to June), daily for 10 days during the summer (mid to late July), and at least once per week during the fall (October to December).
Timing of arrival of adult male NFS
Our hypothesis is that there is no difference among rookeries in the timing of arrival of adult male NFS. NMFS collected first arrival data on St. Paul Island in 2010, and ECO has collected these data since 2016. We will continue to survey the same rookeries and may add representative rookeries from the Lukanin-Kitovi rookery complex, Zapadni rookery complex, Northeast Point rookery complex, and Polovina rookery complex, as appropriate sites are identified for comparisons. Surveys for adult males will occur prior to the arrival of pregnant female NFS.
Index counts of adult female and harem bull NFS
Adult female NFS index counts are part of an important ongoing project that was initiated by MML staff in 2016. In 2017, ECO initiated an index count survey at Reef Gorbatch rookery in order to count adult females and harem bulls on land for a period of 10 days in July. This count provides insight as to what adult females are doing from year to year, it has the potential to be informative about annual population trends where traditional bull counts or pup estimates are useful on a biennial basis. In order to explore the utility of an annual female NFS index count, ECO will continue this survey once daily for 10 consecutive days in mid-July. ECO will also investigate the feasibility of expanding this work to other locations where disturbance is deemed minimal (e.g. Polovina Cliffs).
Timing of departure of adult female and pup NFS
Our hypothesis is that the average date of departure of adult female and pup NFS is later than October 15, the date the rookeries open to the public in the Code of Federal Regulations. The null hypothesis is the average date of departure is October 15. NMFS has flipper-tagged numerous adult female and pre-weaned pups on St. Paul to examine vital rates. Our intent is to take advantage of these identifiable individuals and record presence of tag numbers to determine the timing of departure of those individuals as a representative sample of the population.
Numbers of animals incidentally disturbed during observations are determined using abundance estimates, the proportions of animals in different age/sex categories present (on land) during the surveys and estimates of the proportion of animals present that are disturbed by each activity. These numbers also consider trends in abundance, seasonal behavior and attendance patterns, and the terrain at the specific study locations and associated observation points. These numbers are consistent with estimates for similar activities performed by MML on St. Paul Island and have been reviewed by MML and NOAA Alaska Region personnel.
Project 4. Biological Sample Collection Program (Biosample/Salvage Program)
The purpose of the biosample program is to maximize data collected from dead animals taken during subsistence hunts/harvests of NFS, SSL, and harbor seals in order to gain a comprehensive understanding of population dynamics, vital rates, diets, exposure to diseases, and contaminant exposures and accumulated loads. Collection (salvage) of tissues (e.g. muscle, liver, blubber, blood) and parts (e.g. teeth, vibrissae, stomach and colon contents) from dead animals will ensure long-term availability of marine mammal tissues for a variety of retrospective analysis and enable near real-time processing and interpretation of data to address subsistence-related research questions and concerns. Length, blubber thickness, and/or mass (weight) data are collected annually from subsistence harvested NFS to provide a gross measure of general health of pups and non-pups. Additionally, in order to understand the body condition and health of live non-harvested NFS pups and flipper-tagged non-pups, we request the ability to weigh non-harvested pups and non-pups during the subsistence NFS harvest season (June 23 through December 31).
The objectives of this project are to collect (salvage) tissues and parts from subsistence hunted/harvested and dead stranded marine mammals (pinnipeds and cetaceans), collect condition indices from subsistence harvested and non-harvested NFS such as length, blubber thickness and/or mass (weight), receive tissues/parts from subsistence users, and export hard and soft parts for both research and educational purposes. We believe there is great positive value in getting as much information as possible from subsistence hunted/harvested and dead stranded marine mammals, and non-harvested NFS. ECO seeks to collect tissues, parts, and condition indices from marine mammal species and weight data from non-harvested NFS as a gross measure of general health of pups and non-pups for research via fostering and participating in collaborative partnerships.
ECO has an established co-management sampling program and is in a unique position to collect ongoing local data that can be shared with MML to increase the effectiveness and recovery actions associated with management measures for the eastern Pacific stock of NFS and the western DPS of SSL in the Pribilof Islands, Alaska. ECO has been involved in the collection and distribution of samples from numerous marine mammal species, including but not limited to NFS, SSL, and harbor seals since 1999. Currently, ECO accepts requests from researchers wishing to obtain tissues/parts from subsistence hunted/harvested marine mammals on St. Paul and negotiates a Memorandum of Agreement to establish terms and conditions so that ECO can transfer tissues and/or parts to external researchers (individuals or organizations) to maximize use of available parts. ECO has established agreements for collection of various NFS, SSL, and harbor seal tissues/parts including but not limited to: vibrissae, hair, blood, muscle, teeth, blubber, liver, and bile with some of the following researchers and organizations: Marine Mammal Laboratory; University of Alaska Fairbanks METAL Lab (Dr. L. Rea; S. Crawford); Alaska Department of Fish and Game (Dr. M. Keogh; Dr. Lori Polasek); Alaska Marine Mammal Tissue Archival Project for long-term banking at the National Marine Mammal Tissue Bank; University of North Texas (Dr. A. Roberts; B. Soulen); Duke University (D. Johnston; G. Larsen); University of California San Diego (Dr. C. Kurle); Colorado University (Dr. C. Duncan); and NOAA Polar Ecosystems Program. Additionally, ECO handles new/one-time requests annually.
A key to the success of this program is the already well-established active partnerships. Blubber and liver tissue archival work will continue to be carried out in conjunction with the Alaska Marine Mammal Tissue Archival Project (AMMTAP) and MML. Tissues collected under AMMTAP are archived at the National Marine Mammal Tissue Bank (NMMTB). The NMMTB was established by NMFS Office of Protected Resources, in collaboration with the National Institute of Standards and Technology (NIST). The NMMTB is an important component of the NOAA marine mammal health and stranding response program and is designed to provide cryogenic storage of marine mammal tissues for future retrospective research on animal exposure to emerging contaminants. Since 1987, AMMTAP has periodically collected tissue specimens for NMMTB from the NFS harvests on St. Paul Island and began coordinating these activities with ECO in 1998. More recently, ECO has collected blubber and liver tissues from subsistence harvested NFS from 2014-2019 for AMMTAP. The ECO will continue to collect blubber and liver tissues from subsistence hunted/harvested NFS, SSL, and harbor seal for AMMTAP. Tooth aging and analysis will be carried out in conjunction with MML and Matson’s Laboratory LLC.
Salvage (carcass, tissue, parts)
During subsistence NFS harvests, ECO collects teeth, body length measurements, blubber thickness, mass, blood, blubber, liver, and bile from harvested animals. The Tribal Government has participated in the collection of teeth for research purposes during NFS subsistence harvests since the cessation of the commercial harvests. As a part of our current NFS subsistence harvest monitoring program, ECO collects and ages teeth from harvested animals. The objective of this project is to provide an accurate age determination for all NFS taken during subsistence harvests. ECO also collects teeth from subsistence hunted NFS and SSL and dead stranded marine mammals as possible. Age data from teeth provide information on NFS sampled for ongoing pathology studies as well as additional sample requests and other types of research conducted during harvests. ECO and MML currently independently age all NFS teeth and SSL teeth are sent to Matson’s Laboratory for cementum age analysis. NFS body length measurements are collected in addition to teeth to determine length-at-age. These methods were previously utilized by Dr. A. Trites (1995-1997; 2005-2007) and more recently by ECO (2012-present), and similar data are available from the commercial harvest time period (1911-1984). Length-at-age data, when collected over time, provide insight into factors affecting populations, such as possible food shortage or difficulty of individuals to find sufficient food over time. Stunted growth and reduced body weight are expected in individuals of populations near carrying capacity. It is predicted that the carrying capacity of NFS may be in decline due to food shortage; body measurements collected in the 1990s indicated stunted growth, providing evidence of possible food shortage in foraging areas. Blubber thickness measurements are collected to assess body condition. Collectively, these measurements provide ECO and MML with information on juvenile male NFS body condition. Vibrissae are collected for stable isotope analysis (when funding allows) in order to assess seasonal feeding trends (e.g., feeding over the slope or on the shelf and mean trophic level of prey). Blood is collected for comparative analysis of isotope signatures between the vibrissae and blood and/or archival for future analysis pending additional funding. Blubber and liver tissues are collected for AMMTAP for retrospective research on contaminant levels and animal health.
In 2017, ECO collaborated with Dr. Kathy Burek-Huntington to develop a project to establish baseline polycyclic aromatic hydrocarbons (PAH) levels within NFS via collection of bile from subsistence harvested animals. PAHs are a specific group of chemicals originating primarily from anthropogenic sources that are poorly understood in marine environments (Hellou 2003). The presence of PAHs in marine environments results from discharges from ships, oil seepage and spills, runoff and effluent from land-based industrial processes, and incomplete combustion of fossil fuels (Neff 1990; Baek et al. 1991; Hellou 2003). Individual PAHs have carcinogenic or mutagenic potential and have been investigated with regards to their impacts on marine mammals, including carcinomas observed in SSL after exposure to PAHs (Gulland et al. 1996; Newman and Smith 2006). Potentially devastating impacts of PAHs in pinnipeds include immunosuppression, reproductive failure and functional and morphological changes in thyroid glands and adrenal cortices (Bäcklin et al. 2003). Marine mammals are at risk to PAH exposure because they are highly mobile and utilize waters where the most concentrated levels of PAHs occur. Marine traffic is expected to increase around the Pribilofs in the future as transit through the Arctic becomes more common in general. ECO collected samples in 2017 and 2018 and seeks to continue collections to establish the baseline PAH level of NFS in order to detect potential changes in contaminant levels the future.
Our goal for SSL is to increase local participation in observational data collection and co-management sampling of subsistence hunted and dead stranded SSL on St. Paul. Subsistence hunting for SSL generally occurs between September 1 and May 31. Each year hunters successfully take between 21-35 SSL for subsistence. ECO seeks to work with MML to develop standard tissue collection protocols and sample collection kits for subsistence hunters. ECO will train subsistence hunters to recover a suite of samples to assess diet composition, mercury and contaminant loads, and indicators for disease from known age SSL as feasible. The following tissues/parts may be recovered by hunters and/or ECO: stomach, colon, vibrissae, blubber, liver, hair, blood, muscle, canine and premolar teeth, and ocular fluid. Diet analysis aims to address the hypothesis of whether or not there is an overlap between SSL diet and commercially utilized fish species in the Pribilof region. If there is an overlap, proportions and size lengths of various fish species may indicate the potential impacts commercial fisheries have on the ability of SSL to obtain adequate food in the region. Information on SSL trophic level will also be compared with levels of mercury collected from hair samples. Correlating the mercury load of SSL with proportions of various forage fish and other prey species in SSL diets will provide valuable information about contaminant loads in the Bering Sea pelagic food web and the potential biomagnification of mercury in SSL tissues. We are collaborating with MML and other researchers on stable isotope and mercury analyses. Recent research has shown that young SSL pups in the western Aleutian Islands have higher concentrations of total mercury in hair than at other locations in the Aleutians and Gulf of Alaska (L. Rea, UAF, pers. comm.). This project will sample mercury from non-pups and thereby increase the understanding of mercury exposure in older age classes, health, and reproduction impacts, and aid future management decision making for the Tribal government, Alaska Department of Fish & Game, and MML.
Weighing northern fur seals
The harvest of male NFS pups and non-pups is now authorized from June 23 to December 31 annually. We expect there will be interest from researchers to understand the food safety, health, and biology of harvested (see text above under Salvage) and non-harvested pups. Non-regulatory management measures regarding subsistence use of pups on St. Paul require harvesters to handle and sex all pups to detect and avoid female pups. Since all pups will be rounded-up and handled, we request the ability to weigh harvested and non-harvested pups as an index measure of their condition as feasible. This will provide MML and ECO previously unavailable data on pup growth and condition as they approach weaning. Pup weight data will provide an index of growth and condition over time and will become part of the time series of data on pups and non-pups to enhance MML’s data collected on assessment of demographic parameters. We also request to weigh non-harvested non-pups (flipper-tagged) during the subsistence harvest season to assess condition of known-age NFS as feasible. Weight of flipper-tagged non-pups will contribute to MML’s investigation of juvenile survival rates and body condition.
NFS sample collections (salvaging tissues and parts and weighing) will primarily be conducted during the subsistence harvest season (June 23 to December 31) causing no additional incidental disturbance to animals (in addition to that already exempted for subsistence activities). Numbers of animals incidentally disturbed during collection (salvage) of tissues and parts from other dead animals (subsistence hunted and dead stranded marine mammals (pinnipeds and cetaceans)) are determined using abundance estimates, the proportions of animals in different age/sex categories present (on land) during sample collections and estimates of the proportion of animals present that are disturbed by each activity. These numbers also consider trends in abundance, seasonal behavior and attendance patterns, and the terrain at the specific study locations and associated observation points.
Project 5. Northern Fur Seal Disentanglement Program
The purpose of the disentanglement program is to address the persistent problem of NFS entanglement in derelict fishing gear and other marine debris. The objectives of this project are to: a) improve the survival of entangled NFS on St. Paul by capturing, restraining, removing of debris and releasing them back in the wild; b) track the rate of entanglement as a long-term measure of the success of any efforts intended to reduce NFS mortality due to entanglement; and c) identify the source of entangling debris to better target management efforts to prevent future entanglement. In order to understand how entanglement rates may be changing we intend to use a combination of visual surveys and group round-ups to detect entanglements and, incidental to entangled animals, flipper-tagged animals. When flipper-tagged animals are observed in the round-up we request the ability to weigh those animals as an index measure of their condition, as feasible. These tagged NFS both serve as controls to investigate the effects of entanglement and contribute to MML’s investigation of juvenile survival rates. In addition, entanglements may be observed incidentally during subsistence NFS harvest round-ups, and our intention is to capture, restrain, disentangle, and release those NFS safely, if practical, during the subsistence harvest season. This collaboration with MML will allow us to optimize our efforts into one integrated juvenile male sampling program.
ECO proposes to reduce the level of entanglement and effect the release of NFS from marine debris by performing the following activities: (a) collection of information regarding date, location, sex, age class, debris type, capture attempts, disentanglements, degree of wound, re-sightings, animals sheared, animals with shear marks, scarred animals, and tagged animals and numbers, and (b) calculation of entanglement rates incorporating data from the annual subsistence NFS harvest including debris type, width, mesh diameter, twine size and other information as appropriate.
We have surveyed, observed, and disentangled NFS since 1995, first in collaboration with MML, and independently since 1998. From 1998-2005, observers recorded 795 potential NFS entanglements; 337 captures were attempted; and 282 NFS were successfully disentangled. We estimate that capture attempts are made in response to approximately 40% of the entanglement sightings, in situations when capture conditions are judged to be acceptable based on potential disturbance and safety of the disentanglement team. We estimate 85% of attempted captures are successful, therefore at least 34% of the observed entangled NFS were disentangled from 1998-2005.
When research funding is available we propose to carry out the following: (a) estimate and compare non-pup and pup and male and female NFS entanglement rates and debris composition between St. George and St. Paul Islands both within the breeding season and among years using a non-disruptive observational survey method implemented during 2006; (b) continue to implement the methodology for tracking re-sightings of stranded NFS using a photographic database to verify repeated sightings of individual entangled NFS and better facilitate their eventual capture; (c) identify entangling marine debris removed from NFS for use in debris reduction programs.
ECO may continue or expand the disentanglement program to further address the issue of NFS entanglement and progress toward preventing future entanglement by addressing the source of marine debris. The ECO has been an active participant of the Alaska Marine Mammal Stranding Network (MMSN) since 1999. We have a current 3-year Marine Mammal Stranding Agreement with NMFS, which authorizes Dr. Divine, Ms. Lestenkof and other delegated ECO representatives to take species of marine mammals under the Marine Mammal Protection Act (MMPA) for the purpose of dead animal investigation and response, including the collection and transfer of animal parts. Under this agreement the ECO may also take marine mammals species covered under the MMPA for the purpose of live-stranding first response, beach triage, beach release, temporary holding for assessment and triage, translocation and/or transportation to an authorized rehabilitation center but does not authorize any projects involving “intrusive research”. The MMSN stranding agreement authorizes ECO to capture and disentangle NFS but does not authorize incidental disturbance resulting from disentanglement captures and intentional harassment caused by the group round-up technique used to capture and disentangle more than 1 seal in the same group.
Numbers of animals incidentally disturbed during directed disentanglement activities are determined using abundance estimates, the proportions of animals in different age/sex categories present (on land) during disentanglement activities and estimates of the proportion of animals present that are disturbed by each activity. These numbers also consider trends in abundance, seasonal behavior and attendance patterns, and the terrain at the specific rookeries where the activities are conducted and associated observation points.
The number of male pups and non-pups that will be intentionally harassed during group round-up technique are maximum estimates determined previously by P. Zavadil in consultation with B. Robson and S. Insley, who have collectively conducted similar procedures over the past three decades. Dr. Insley participated in disentanglement roundups during the late 1980s to early 1990s. Mr. Robson has been involved in entanglement surveys and captures since 1990, first with the MML and after 2002 under the authority of NMFS Permit No. 1066-1750-00, held by M.T. Williams. The harassment numbers are also consistent with Permit No. 1066-1750-00 and have been reviewed by MML and Alaska Region staff.
Projects 1-5. Unintentional Mortalities
Unintentional mortality requests are included in the Project Supplemental Information: Lethal Take section and its associated Supplemental Information file.

II.	PROJECT DESCRIPTION
Project 1. Marine Mammal Population Monitoring/Assessment
Aerial survey with sUAS (Survey, aerial; Remote vehicle, aerial [fixed wing, VTOL]; Photograph/Video): Aerial surveys will be conducted annually with sUAS (fixed wing, rotary, or hybrid sUAS platforms that are capable of VTOL or assisted horizontal launch) equipped with cameras and/or sensors (e.g., thermal, RGB) to survey and photograph harbor seals, SSL, and NFS in the Pribilof Islands to obtain pup and non-pup counts. The ECO will use small UAS weighing less than 55 pounds, as categorized by the Federal Aviation Administration (FAA). Payload components will be dependent upon the scientific mission and will be appropriate for the platform. A variety of survey types may be employed with the focus to be on photogrammetry of pinnipeds. Survey efforts will focus on collecting data (imagery) to calculate pinniped trend and abundance estimates. Data collected may also include thermal and color imagery. The number of surveys per year will depend on the scientific needs, weather, and platform availability. The sUAS will never be beyond the line of sight and will have an auto-return feature should the device fail, as required for FAA operation. Remote Pilot in Command will always possess necessary and appropriate FAA permits and authorizations including a valid Remote Pilot Certificate.
The ECO will conduct land and boat-based sUAS operations to collect aerial imagery of pinniped rookery and haulout sites. Flights will be conducted above a minimum of 150 feet above ground level in order to provide the necessary photographic resolution to discern pups, comply with FAA regulations, and to ensure minimal levels of disturbance. It is necessary to obtain vertical photographs (to fly directly over animals) to ensure the highest quality images for enumeration and photogrammetry. ECO has built a history of safe operations with no take during sUAS operations since 2016. Additionally, in July 2015, MML examined takes of harbor seals on Otter Island relative to a sUAS flying along shore at various altitudes (MML, unpub. data). Based on their examination there was no visible reaction by the seals to the sUAS at any altitude, and it was determined that UAS could be a valuable tool for monitoring harbor seals in the Pribilof Islands when compared to shore-based observer counts. ECO currently owns a fleet of DJI platforms that have a max flight time of approximately 30 minutes and has established Standard Operating Procedures (SOP) for safe land- and boat-based operations.
Boat-based sUAS surveys will be conducted from the SAR/V Lunax? (SAFE Boat) or similar vessel at a minimum distance of 500 yards from shore of any focal areas. It may be necessary to circumnavigate Walrus and St. George Islands to visually search for target animals hauled out on shore. If target animals are spotted, the sUAS will be flown over animals to collect aerial imagery using either remote pilot in command piloted flights or pre-planned flight missions for standardized transects. The sUAS will be vertically launched, flown, and recovered by hand from the Lunax? using safety procedures detailed in the ACSPI SOP. This will minimize disturbance to animals caused by landing on shore. For remote pilot in command piloted flights the pilot will attempt to fly slow (<5 m/sec; <10 miles/hour) straight line transects to reduce disturbance take. Photos will be taken at regular intervals during the flight or while hovering, with a minimum of 75:80 front:side overlap to produce sufficient overlap to stitch images together in post-processing. For standardized transects, targeted pinniped waypoints will be pre-programmed into a flight plan using GS Pro or similar flight planning software.
If we cannot launch the sUAS from the Lunax? a suitable ground-based launch site downwind and away from target animals will be used to minimize disturbance. Up to 10 fabric targets (measuring 48x48 inches) may be placed on the ground to act as control points. Spotters will be placed appropriate to the FAA licenses and authorizations.
Aerial images resulting from all surveys will be processed and analyzed following the same methods and protocols used by MML. Images and count data will be shared with MML for photogrammetry and stock assessment reports. MML will in turn provide count data to ECO to compare and validate ECO counts on the Pribilofs, so that ECO can build capacity to conduct this type of work. ECO has the appropriate FAA permits/authorizations to conduct this work. Two ECO/ NMFS-approved personnel for Permit No. 19436-02 (L. Divine and A. Lestenkof) obtained their FAA Part 107 Remote Pilot Certificates in 2016, and L. Divine has renewed her RPC in 2020 (expires 2022). A. Lestenkof will recertify as soon as is possible in light of COVID-19 state mandates and local travel restrictions. All pilots will hold current valid certificates during implementation of these activities for the duration of this permit (2021-2026).
There will always be a visual observer present looking for aircraft and at least one experienced observer knowledgeable of pinniped behavior will monitor animals for any reaction to the UAS. If pinniped disturbances are observed during a flight, the UAS will be directed to a greater altitude or distance from the animals; the flight will be discontinued if animal reactions to the UAS do not subside. On the Pribilof Islands, up to 80 pup harbor seals and 200 non-pup harbor seals (both sexes); up to100 pup SSL and 1,000 non-pup SSL (both sexes); and 100 pup NFS and 5,000 non-pup NFS (both sexes) will be disturbed during up to10 surveys, annually. Additionally, ECO will use sUAS to monitor the rate of accumulation of marine debris on cleaned rookeries and haulouts on St. Paul Island. ECO will fly shorelines where marine debris removal activities occur once a month as weather permits, as well as a control beach close to the town of St. Paul where removal activities do not take place and seals are rarely hauled out. Up to 100 pup NFS and 1,000 non-pup NFS of either sex will be disturbed (collectively during up to 12 surveys, once per month) annually.
Project 2. Northern Fur Seal Disturbance using VHF
For the purpose of investigating adult female NFS responses to subsistence and research activities at NEP on St. Paul Island, as many as 100 non-pup female NFS will be captured, restrained, weighed, and marked (flipper tags) on St. Paul Island annually. Captures at NEP will take place between September and October. One capture per animal is sought for this project. Incidental disturbance resulting from non-pup captures is estimated to occur for up to 200 pup NFS and 2,100 non-pup NFS of either sex annually.
ECO will deploy up to 7 VHF receiver stations (Advanced Telemetry Systems, Isanti, MN, USA) annually to automatically detect tagged NFS presence/disturbance throughout the 2021-2026 seasons in collaboration with NOAA Fisheries Alaska Region and MML. The NEP study area will consist of at least 7 receiver stations at Morjovi and Vostochni Rookeries (approximately 3 stations at Morjovi and 4 stations at Vostochni). All stations will be outfitted with omni-directional antennas. Range-testing in 2019 established 10-11 sites, and these sites were reduced to 7 semi-permanent sites with antennas mounted to tripods. The final site choices are based on maximizing coverage of the rookeries while also minimizing disturbance. The placement of semi-permanent stations will allow 24/7 searches to be conducted for tagged NFS. For each station, a scanning receiver will continuously scan according to a pre-programmed cycle through 8 or 9 frequencies when running in “Aerial Mode”. The receiver will be housed in a yellow drum/barrel that is designed to support an omni-directional GPS antenna, a marine grade battery, accessory cords, and a solar panel. A new, 10-year reference transmitter (pulse-coded) will be placed in a location central to both rookeries to continuously monitor operational status of all receiver stations. Receivers will be deployed annually during the spring before the arrival of NFS and removed in the winter after departure of NFS; there will be no incidental disturbance resulting from receiver deployment or removal on the rookeries.
ECO will download data recorded on receivers monthly (July through December). All data will be duplicated and stored at two locations in ECO on St. Paul Island, as well as submitted to MML. Incidental disturbance resulting from monthly data downloads and station maintenance is estimated at 200 pup NFS and 2,000 non-pup NFS of either sex annually.
Tag resighting observations will be conducted as necessary to verify the animals with VHF tags (e.g., if one of the tags fail). Tag resighting observations will be conducted on up to 6 occasions annually at the study sites and may occur during the pupping season, late summer, and in the fall. These observations are conducted from locations overlooking the rookeries or catwalks where possible and may be conducted by moving among juvenile and subadult male aggregations on rookery and haulout areas as needed. Tags will be read by individuals using binoculars, spotting scopes, telephoto cameras, and visually, as well as radio-tracking and radio-logging equipment and may result in the incidental disturbance of up to 300 pup NFS and 3,000 non-pup NFS of either sex annually.
Detailed methods for Project 2 procedures are as follows:
Adult captures (Capture, other): Up to 100 non-pup females will be captured using a hoop net or a noose pole as described by Gentry and Holt (1982). Hoop nets are preferred for individual captures, particularly those that occur after the beginning of August when the territorial structure of rookeries begins to break down, while noose poles are generally used when groups of animals are rounded up for capture, as well as for individual captures that occur earlier in the season (before August). The body of a hoop net is 1/4-inch mesh and the head cone is double-layered netting of 1/8 inch mesh. Noose poles are typically 4 cm x 4 cm (2”x 2”-dimension lumber) wood cut to varying lengths. Corners are beveled to an octagonal shape, and a 1 cm diameter, 95-110 cm long rope is threaded through two holes drilled 15-23 cm apart near one end of the pole to create a loop 45-50 cm deep. Researchers approach the rookery, keeping a low profile (low walk or crawl) and moving slowly. Small groups of NFS may be isolated when larger sample sizes are required. The group is maintained by placing individuals in strategic locations around the NFS, and we attempt to include 10-30 adults in the group, resulting in much less overall disturbance (per animal captured) than individual captures. The hoop net is placed over the NFS's body, or the noose is placed over the NFS's head and the pole is rotated until the loop tightens around the neck; both methods allow a single person to control seals weighing up to 60-65 kg. Two people then carry the NFS to a safe location. While the NFS is being moved, its condition is closely monitored to detect signs of stress. NFS adult females are then physically restrained on a restraint board (a wood platform with a wooden yoke that closes around the neck). "V" notches are cut in the yoke and lined with foam. The downward pressure provides restraint and prevents the seal from turning its head side to side. The upper blade of the yoke is tied into place with a quick-release knot, and pressure adjusted such that restraint is achieved without restricting blood flow or airway. Additionally, the depth of the lower "V" notch helps to prevent obstruction of the airway. A neoprene wrap is placed around the fore flippers and fastened with Velcro bands to restrict fore flipper movement. During processing, each animal is attended by at least 2 researchers (1 to monitor the animal and 1 to weigh and record data), and 2 animals are often processed simultaneously. Handling times typically average about 10 minutes for marking and weighing activities.
Weigh: The physically restrained animal is secured in or on an apparatus (e.g. bag, harness, net, bucket, brailer bag, stretcher, sling, kennel, restraint board) and suspended from a mounted (e.g. on a bipod, tripod, boom, pole, plank) electronic or spring scale to weigh, then gently returned to the ground. Restrained or controlled animals may also be weighed using a platform scale. The number of personnel used will be based on the number that are necessary to ensure safety of animal, people, and accurate weighing. Personnel involved in weighing will monitor animal orientation to ensure an open airway, prevent injury, and minimize discomfort (e.g., due to flipper, head, or neck position, damage to teeth caught on the weighing implements, etc.).
Mark, flipper tag: Animals are marked with two tags, one in each flipper. One VHF flipper tag and one plastic or metal identification tag. VHF flipper tags (Advanced Telemetry Systems, Isanti, MN, USA) will be used for information on attendance. The technology in this field is changing quickly and it is impossible to know the size and weight of instruments that will be deployed over the duration of the permit. However, the distinct trend has been for instruments to become progressively smaller and lighter as micro processing boards have decreased in size over time. Currently, weight of individual VHF flipper tags are 162 g and footprint is less 4 cm2, however, these sizes are subject to change and may be lighter or smaller than described. VHF flipper tags will be applied to a hole in the flipper, “wrapping” the tag through the hole, and securing the open end with a small screw so the entire plastic tag is closed loop (i.e., they can only fall off through breakage or complete ‘tear’ in the flipper). More recent VHF flipper tags are potted in epoxy resin cast as a female tag and paired with a male Allflex disk tag. These are labeled and can often be read when the opposite tag is not showing. They are always paired with an Allflex narrow sheep tag in the opposite flipper. Individuals that are outfitted with a VHF flipper tag will also receive an Allflex flipper tag attached to the trailing edge of the NFS’s fore flipper, near the hairline where it meets the body on the opposite flipper that the VHF flipper tag is attached to. We will utilize Allflex narrow sheep tags as they are the most common tags used presently (as described below).
Tags used for visual identification possess a unique number, letter and/or color combination on the surface of the tags and are typically commercially-available livestock tags with piercing posts made of hard or soft plastic (e.g. Allflex, Dalton, Duflex, Ketchum, Temple), or a metal alloy (e.g. Monel) in a variety of shapes. Visual identification tags are generally less than 6 cm x 6 cm in size, and less than about 10 g in mass.
Flipper tags will be attached to the proximal trailing edge of the fore flippers. Caution will be used to avoid the digital bones and blood vessels and to ensure the post of the tag is longer than the thickness of the flipper at the tag site. The tag should be applied so that the sharp tip of the post faces outwards, rather than against the animal's body. Tags will be applied using manufacturer-specified applicators. In some cases, the flipper will be secured while a small hole or incision is made prior to tag insertion to improve tag application efficiency. Small holes or incisions will be created using a sterile biopsy punch, sterile scalpel blade, or cleaned and disinfected leather punch. Tissue pushed free from the flippers during insertion of the tags, or as a result of flipper punch prior to tag application, may be retained for use in other studies. Each animal may receive one or two flipper tags (one in each fore flipper) as appropriate to meet study objectives. Flipper disfigurement or signs of previous injuries may preclude tagging one or both flippers or may slightly alter tag placement.
Attachment of single-post flipper tags should use as much aseptic technique as is practicable within the study design. At a minimum, all tag attaching equipment will be cleaned and washed thoroughly with soap and water, then disinfected (e.g. disinfecting solutions such as bleach, alcohol, chlorhexidine, accelerated hydrogen peroxide) between use among different animal groups (e.g. unique rookeries or haulouts). When possible, tag application sites also may be cleaned with topical antiseptics (e.g. alcohol, povidone-iodine, chlorhexidine, or other surgical preparation solutions). If cleaning or disinfecting the tag attachment site is infeasible, the jaws of the tag applicator (with tag inserted) may be dipped into or wiped with an antiseptic or disinfectant solution prior to applying tag into flipper webbing.
Flipper tags may be lost, broken or excessively worn over time. Tags may be re-applied to pinnipeds whose tags have been lost or are no longer readable in order to maintain the individual identities of these animals. Tag attachment site and tag retention will be documented whenever possible during resights in order to assess healing, tag migration, tag wear and flipper condition. Other marks and tags may be used in conjunction with flipper tags to meet study objectives.
Project 3. Northern Fur Seal Habitat Use Studies
Counts of adult male, adult female, harem bull, tagged adult female, and tagged pup NFS will be conducted annually by observers overlooking the rookeries from clifftops, fixed blinds, catwalks, and other remote vantage points and do not entail the capture of NFS or major disruption of rookeries. Research protocols for NFS habitat use studies on the Pribilof Islands call for weekly (at least once per week) observations at the study sites. Researchers keep low profiles and move slowly along the edge of the rookery to access the observation points and conduct direct counts either with the naked eye, binoculars, spotting scopes, or telephoto cameras.
Detailed methods for surveys conducted as part of Project 3:
Timing of arrival of adult male NFS
Adult male counts are conducted in the spring/summer (late April to mid-June) prior to the arrival of adult females and the pupping season. ECO will survey rookeries to identify and count adult male NFS on land and the timing of arrival on an annual basis. We will follow the protocol developed by ECO using the survey methods described in Williams et al. (2011) to identify the timing of first arrival of adult male NFS on St. Paul. Based on the results of Williams et al. (2011) we will start surveying rookeries in late April and will survey at a consistent frequency (at least twice per week) to enable statistical analysis of adult male NFS count data. Count data will be collected by rookery section. Surveys are discontinued when the arrival of females on land make adult male counts impossible. The date of first arrival will be defined as the day on which 1% of the maximum number of adult males on a given rookery section have arrived. NMFS annually counts adult males as an index for population production (Antonelis 1992). The NMFS adult male (bull) count occurs annually during the second week of July and estimates the maximum number of bulls on island (Gentry 1998). ECO will initially use the NMFS bull count from the previous year to estimate the 1% threshold for the present year. After NMFS has completed the bull count for the present year, timing of arrival data will be compared using both the bull count data from the previous year and the same year to determine variability in the timing of first arrival for the year as determined by ECO. These surveys are conducted from remote vantage points with binoculars or spotting scopes and should not result in any temporary displacement of animals.
Index counts of adult female and harem bull NFS
Adult female and harem bull counts are conducted in the summer (mid to late July) for a period of 10 days during the pupping season on an annual basis. Island Sentinels will survey Gorbatch Rookery to count harem bull and adult female NFS on land. These data will be stored in the BeringWatch database and shared with NMFS (R. Ream) for future collaborations. These surveys are conducted from fixed blinds with binoculars or spotting scopes and should not result in any temporary displacement of animals.
Timing of departure of adult female and pup NFS
Adult female and pup counts are conducted in the fall/winter (October to December) after the pupping season. ECO will survey the departure of northern NFS by counting the number of tagged adult females and pups still occupying the rookery at a consistent frequency (at least once per week) from October 1 until tagged NFS depart the island on an annual basis. Adult females and pre-weaned pups have been previously tagged at Polovina Cliffs rookery. We will also count the number of dead tagged females and pups observed and record the tag number for MML. ECO will survey Polovina Cliffs rookery until December 1 or until the number of tagged animals occupying the rookery is zero. Tags will be read with binoculars, spotting scopes, or telephoto cameras and may result in temporary displacement of animals. The method of taking may occur from the sight, scent or sound of human presence. 
Incidental disturbance resulting from all surveys is estimated at 2,000 pup NFS and 4,000 non-pup NFS of either sex. These numbers consider trends in abundance, seasonal behavior and attendance patterns, and the habitat at the specific study locations (focal study rookeries, other rookeries, haul-outs) and associated observation points.
Project 4. Biological Sample Collection Program (Biosample/ Salvage Program)
ECO plans to collect (salvage) tissues and parts from subsistence hunted/harvested and dead stranded marine mammals (other pinnipeds and cetaceans), receive tissues/parts from subsistence users, and to export hard and soft parts for both research and educational purposes.
Authorization is requested for the collection and export of scientific material including hard tissue (skulls and skeletons), teeth, soft tissues and/or whole carcasses from these species for both research and educational purposes. Sample collections from subsistence hunted/harvested NFS, SSL, and harbor seals and dead stranded marine mammals will occur year-round. The export of tissues would occur on a quarterly or as needed basis depending on the timeframes of individual collaborating researchers. We have not identified any foreign laboratories or researchers that we would export samples to, but we have worked with researchers at the North Pacific Universities Marine Mammal Research Consortium and Pacific Identifications, Inc. in B.C., Canada.
ECO works collaboratively with MML to develop standard data and tissue collection protocols to assess diet composition, mercury and contaminant loads, and to serve as an indicator for disease from known-age pinnipeds. Body length measurements, blubber thickness, mass, teeth, vibrissae, hair, blood, muscle, ocular fluid, blubber, liver, and bile will be collected from up to 400 NFS, 100 SSL, and 50 harbor seals (of all age classes and both sexes; subsistence harvested/hunted or dead stranded animals) on St. Paul. Other samples may be added to the protocol over time and requested from hunters/harvesters. Samples from up to 10 dead stranded cetaceans and 10 other pinnipeds will also be opportunistically collected and maintained when animals found are dead during other research activities on St. Paul Island.
Incidental disturbance to peripheral animals while collecting samples from subsistence hunted and dead stranded marine mammals may occur on each occasion of sampling but are expected to be low. NFS sample collections will primarily be conducted during the subsistence harvest season (June 23 to December 31) causing no additional incidental disturbance (in addition to that already exempted for subsistence activities). Incidental disturbance of NFS, SSL, and harbor seals will primarily occur during sampling of hunted and dead stranded animals. However, such cases are expected to be infrequent based on historical subsistence takes of hunted marine mammals and stranding events. Furthermore, the only location where SSL haulout in high numbers are on Sea Lion Rock. Similarly, harbor seals only haulout on offshore rocks at Little Zapadni and Southwest Point on St. Paul Island and on Otter Island.
For NFS, we estimate incidental disturbance of up to 2,000 pups and 6,000 non-pups of either sex during sampling on St. Paul Island annually. We estimate incidental disturbance of up to 50 pup SSL and 1,000 non-pup SSL of either sex during sampling on St. Paul Island annually. For harbor seals, we estimate incidental disturbance of up to 80 pups and 200 non-pups of either sex during sampling on St. Paul Island annually. These numbers consider trends in abundance, seasonal behavior and attendance patterns, and the habitat at the specific study locations (focal study rookeries, other rookeries, haul-outs) and associated observation points.
Detailed methods for Project 4 procedures are as follows:
Salvage (carcass, tissue, parts): We will collect body length measurements, blubber thickness, mass, teeth, vibrissae, hair, blood, muscle, ocular fluid, blubber, liver, and bile from dead animals to provide information regarding age, general body condition, and health of the animals.
Body length, blubber thickness, mass, teeth, and vibrissae collections
ECO will measure the body lengths of harvested NFS using a metal caliper placed over each animal, measuring from the tip of the nose to the tip of the tail to the nearest centimeter. Blubber thickness will be measured with a small plastic ruler to the nearest millimeter where the chest is cut open for exsanguination. Harvested NFS pups will be weighed using a platform scale or in or on an apparatus (e.g., bag, net, bucket, brailer bag, sling) and suspended from a mounted (e.g. on a bipod, tripod, boom, pole, plank) electronic or spring scale to weigh.
Measurements will be recorded in a field notebook with each assigned a unique sample identification number, harvest date, and harvest location. A zip lock bag labelled with the corresponding sample number will be inserted into the animal’s mouth, ensuring canine teeth that are collected after butchering is complete are matched with the correct length, blubber thickness, and mass measurements. Once the seals are butchered, we remove the snout with upper canine teeth intact and vibrissae from the heads that have zip lock bags. Snouts will be inserted into the numbered bags and frozen for storage prior to boiling and extracting the upper canine teeth. Teeth are processed in a steam kettle to extract the upper and/or lower canine teeth periodically throughout the harvest season. At least two of the longest vibrissae from each snout will be collected and placed in labeled whirl-pak bags when snouts are boiled. Canine teeth will be visually inspected for growth rings to estimate the age of individual animals following Scheffer (1950) and will be paired with body lengths for each individual to assess length-at-age. Vibrissae will be collected following the sampling protocol that was developed in collaboration with MML and archived in St. Paul for future stable isotope analysis.
Tooth collections
Teeth will be collected from subsistence hunted/harvested and dead stranded animals found on the rookeries to determine the age and sex of these animals. Teeth from NFS taken in the subsistence hunting and harvest seasons will be collected and maintained in collaboration with MML to continue the historic data set of samples from subsistence harvested animals taken on St. Paul Island. For NFS, we will collect the upper and/or lower jawbones from each subsistence hunted or stranded NFS and from at least 50% of harvested NFS to extract up to four teeth per animal. ECO will collect upper canine teeth (n=2) and/or lower canine teeth (n=2) from hunted/harvested NFS. For SSL, we will collect the upper jawbone from each subsistence hunted or stranded SSL to extract up to four teeth per animal. ECO will collect both upper canine teeth (n=2) and two premolar teeth (n=2) from all dead animals. For harbor seals, we will collect the upper and/or lower jawbones from each subsistence hunted or stranded harbor seal to extract up to four teeth per animal. ECO will collect upper canine teeth (n=2) and/or lower canine teeth (n=2) from all dead animals. If additional requests are made by other researchers for teeth from subsistence hunted/harvested animals, these requests will be handled through the St. Paul Island Co-management Council.
The age of pinnipeds will then be determined by counting incremental growth layers on a longitudinal section of the tooth, using standard procedures developed by MML. Teeth will be retained by ECO for archival. Teeth may be exported to Matson’s Laboratory for cementum age analysis and returned to ECO for archival. All age data from subsistence hunted/harvested animals for which samples were collected will be entered into the ECO BeringWatch database and shared with other researchers that may have received samples from the same animal and with MML for use in other studies.
Vibrissae, hair, blood, and muscle collections
Vibrissae, hair, blood, and muscle samples will be collected and shipped to a lab (e.g. UAF) for stable isotope analysis and duplicated for mercury analysis. Two of the longest vibrissae from one side will be pulled out and stored in whirl-paks. Hair samples will be collected down to the skin over a 2 x 2 in area on the right flank using grooming scissors and stored in whirl-paks. At least one 8.5 mL tube of whole blood will be collected, centrifuged on island and separated into serum/plasma and red blood cells, and stored frozen. A small piece of skeletal muscle tissue (minimum 4 cubic cm) will be collected from a standardized region on each animal (to be determined) and stored frozen in whirl-paks prior to analysis. All samples except vibrissae and hair will be stored at -80ºC on island until they are shipped to a lab for analysis.
Blood collections
Blood will be collected as whole blood samples from subsistence hunted/harvested animals. At least one 8.5 mL tube of whole blood will be collected. ECO personnel will further process the blood samples by centrifuging in blood vials designed to separate the red blood cells from the plasma components. Advantec Nobuto Blood Filter strips will be used to test for the presence of antibodies in the blood serum and serve as an indicator for disease (Dusek et al. 2011, Hoover-Miller 2013). Strips hold approximately 0.1 mL of blood or 0.04 mL of serum. Strips can be analyzed, stored, and shipped without refrigeration.
Blubber and liver collections
ECO personnel will remove 300-350 g samples of blubber and liver from each subsistence hunted/harvested animal. Blubber and liver samples will be placed in separate, sterile Teflon bags and processed immediately after collection in the NOAA laboratory facility following the Tissue Sampling Protocol for the National Marine Mammal Tissue Bank (Becker et al. 1999). Samples will be labeled, stored in a -80 ?C freezer on island, and then packaged into Teflon jars and shipped in a dry shipper to Hollings Marine Laboratory in Charleston, South Carolina, for long term storage in the NMMTB through AMMTAP. A minute portion of liver tissue will be cut away from the larger portion collected, frozen, and shipped to a lab for stable isotope and mercury analysis.
Bile collections
ECO personnel will remove up to 10 mL of bile from intact gallbladders of harvested non-pup male NFS as time allows during the harvest season. Bile samples will be collected with a 15-mL syringe into amber vials after livers are removed during butchering. Bile may be pooled from multiple NFS during one harvest event. All samples will be labeled with the date, harvest location, and number of individuals contained in the sample. Vials will be wrapped in aluminum foil, transported to ECO and stored at -80?C for future baseline contaminants analysis. These data will enable future comparison of PAH levels over time and may provide information regarding changes in exposure or accumulation rates in NFS. Increased exposure or accumulation of contaminants, including PAHs, is a concern of subsistence users who depend on seal meat as a consistence source of protein.
Weigh: We will collect weights as a measure of health and condition of NFS pups and non-pups during subsistence harvest activities. Non-harvested pups and non-pups (flipper tagged) will be physically restrained and secured in or on an apparatus (e.g., bag, harness, net, bucket, brailer bag, stretcher, sling, kennel, restraint board) and suspended from a mounted (e.g. on a bipod, tripod, boom, pole, plank) electronic or spring scale to weigh, then gently returned to the ground. The number of personnel used will be based on the number that are necessary to ensure safety of animal, people, and accurate weighing. Personnel involved in weighing will monitor animal orientation to ensure an open airway, prevent injury, and minimize discomfort (e.g., due to flipper, head, or neck position, damage to teeth caught on the weighing implements, etc.).
For the purpose of estimating pup survival, as many as 400 pup NFS of either sex will be handled during subsistence harvests, restrained, and weighed annually. Pups will be handled to record their weight during group “round-up” technique used to capture seals for subsistence causing no incidental disturbance. Management measures regarding the harvesting of pups on St. Paul require harvesters to handle and sex pups to detect and avoid female pups. Pups that are rounded-up and handled will be weighed as described in Project 2. Non-harvested pups will be carried to a safe location near the harvest area and restrained by hand or with a neoprene wrap placed around the fore flippers and fastened with Velcro bands to restrict fore flipper movement. The pup will be constantly monitored for signs of stress. Multiple pups may be weighed at one time to minimize additional disturbance. Animals will then be carried and released close to the harvest location.
For the purpose of estimating non-pup NFS survival, as many as 100 tagged non-pups of either sex will be handled during subsistence harvests and disentanglement activities, restrained, and weighed annually. Non-pups will be handled to record their tag number and weight during group “round-up” technique used to capture seals for subsistence and disentanglement activities (as described in Project 5) causing no incidental disturbance during subsistence activities. We estimate that 50 non-pups will be weighed during subsistence activities and 50 non-pups will be weighed during disentanglement activities. Incidental disturbance of pups and non-pups while capturing and weighing non-pups during disentanglement activities are included in the totals for the Disentanglement Program (Project 5). Tagged non-pups that are rounded-up and handled will be weighed as described in Project 2.
It is not possible to list additional laboratories or researchers that will request samples in the next five years. Each of the previously listed researchers and organizations above that we have collaborated with have multiple research programs, many of which may request samples. ECO will collect, receive, and export hard and soft parts from subsistence hunted/harvested and dead stranded NFS, SSL, and harbor seals for potential researchers and research projects on a case-by-case basis, in consultation with the NMFS permit office. Additionally, we will collect, receive, and export hard and soft parts from unidentified dead stranded cetaceans and other pinnipeds, consistent with the provisions of our Alaska Region Marine Mammal Stranding Agreement.
All parts and specimens will be collected in the field as fresh as possible, usually within 24-48 hours of a harvest, hunt, or stranding event. Samples will be placed in one of the following containers: a plastic/glass/Teflon canister, jar or vial, plastic whirl-pak, zip-loc, or heavy-duty trash bag.
Each sample is assigned an accession number once received at the ECO and entered in the BeringWatch Database. Accession numbers facilitate the sample processing by first recording date of entry and associated documentation surrounding the sample, such as: date of collection, species or common name, general harvest location, sex, listing of the samples in the sample, and collector provided remarks.
All samples except processed teeth, vibrissae, hair, cleaned skulls, skeletons, and bones will be stored in containers or bags in freezer conditions at -80ºC, unless specified and agreed upon by the ECO and collaborating researcher.
All samples except teeth, vibrissae, hair, cleaned skulls, skeletons, and bones will be packed and shipped in leak proof containers, such as ice chests or Rubbermaid containers. Frozen samples will be shipped frozen in leak proof containers with ice packs or dry ice, unless specified and agreed upon by ECO and the collaborating researcher. All samples will be shipped with a copy of the biological samples collection forms and a copy of both parties’ current scientific research permits. The ECO will notify the receiver of shipment at least 24 hours in advance to make sure parties available to receive shipment. On one side of the outer package will include the Addressee, Addressor, and “this package conforms to 49 CFR 173.4”.
Project 5. Northern Fur Seal Disentanglement Program
Pups and non-pups will be captured, restrained, and disentangled when detected during observational surveys, subsistence harvests, or during directed research group round-ups. Disentanglement captures of NFS on rookeries during the breeding season are currently not conducted. ECO will initiate any program to attempt pup/non-pup rookery captures in close consultation with NMFS co-management partners including Protected Resources Division staff for the Pribilof Islands. From 1998-2005, observers recorded 795 potential NFS entanglements; 337 captures were attempted; and 282 NFS were successfully disentangled. Based on recent years (post-2002) the actual number of non-pup capture attempts will likely be less than 100 NFS annually. Disentanglement captures of NFS are currently covered under our Marine Mammal Stranding Agreement with NMFS. However, incidental disturbance caused by disentanglement captures, observational entanglement surveys, and intentional harassment caused by the group round-up technique used to capture and disentangle more than 1 seal in the same group are not covered in our Stranding Agreement and are included below. 
For the purpose of estimating entanglement rates for different age and sex categories of NFS, we will use a combination of observational entanglement surveys and group round-up surveys of rookeries and haulouts to detect and respond to entangled pup and non-pup NFS of either sex on St. Paul Island. Observational entanglement surveys will occur weekly from May to December. During observational surveys, researchers use binoculars to count entangled and non-entangled NFS at sampling sites on rookeries and haulouts. Specific sites are selected for repetitive sampling of pup and non-pup NFS of either sex from cliff tops overlooking rookeries and haulouts. Based on prior work conducted on St. Paul Island in 2004-2005 (Zavadil et al. 2005, 2006) non-cliff top sites do not result in reliable and balanced detection of entangled and non-entangled NFS. Sites are selected to ensure that the viewing location is reliable and to avoid disturbance to adjacent NFS during most wind conditions. NFS observational entanglement surveys will also take place during the subsistence harvest season (June to December). Entanglement surveys conducted during the harvest season will involve no additional disturbance in addition to that already permitted for subsistence activities. There are no intentional takes of NFS associated with observational entanglement surveys. However, we estimate up to 2,000 non-pups of either sex may be incidentally disturbed cumulatively for each year on haulouts during weekly observational entanglement surveys.
Directed research group round-up surveys will be conducted at selected study sites during the summer (June-July) on hauling grounds, where non-pup male NFS are located reliably due to their life history. When flipper-tagged animals are observed in the group round-up we will weigh those animals as an index measure of their condition as feasible. The group round-up method involves surrounding a group of male NFS (e.g. maximum of 500) to the extent that the target NFS (i.e. entangled or tagged) are contained within the group. The process is the same as Fowler (1987) where it is described in detail. While the NFS are contained, they are allowed to spread out to avoid stress, over-heating, and excessive movement. NFS are allowed to escape through a controlled exit (i.e. bottleneck) point where the target (i.e. entangled or tagged) animals can be readily spotted and captured. When a target fur seal exits, the bottleneck can be quickly closed so that any remaining targets are not simultaneously lost. This process is continued until all target animals are captured, restrained, and disentangled. For flipper-tagged NFS, we will capture, restrain, and weigh up to 50 non-pup NFS of either sex to record their tag number and weight (as described in Project 4) annually. Takes of NFS associated with group round-up technique will include intentional harassment of up to 5,000 male NFS (all life stages including weaned pups and older) annually. In addition, because SSL are infrequently found at most NFS hauling grounds, up to 80 SSL (of all age classes and both sexes) may be incidentally disturbed during this research activity annually.
Debris removed from entangled NFS will be cleaned, characterized, and retained by ECO. Entangled NFS not disentangled will be photographed and their debris characterized into consistent groups based on characteristics of the debris that can be reliably detected. The type and weight of debris, mesh size or circumference of entangling loop, and the extent of the wound created by the debris is recorded. Debris is catalogued and retained by ECO for use in identifying the source of the debris. There is no incidental disturbance associated with this activity. 
Detailed methods for Project 5 procedures are as follows:
Disentanglement crew members search for entangled NFS during observational or round-up surveys at specified trend sites. In addition, non-survey areas are regularly scanned for entangled NFS and entanglement surveys will also be a regular part of subsistence harvests. Regular surveys are completed from vantage points overlooking the rookeries and haulout areas, using spotting scopes and binoculars to scan the area for entangled NFS. The public and other researchers may observe entangled NFS and report them to the ECO for response. Once entangled NFS are detected we evaluate whether the circumstances allow us to independently attempt to capture the NFS or round-up the entire group of nearby NFS including those entangled. Independent captures allow us to improve the survival of individual entangled NFS but does not allow us to capture multiple “target” (i.e. entangled or tagged) NFS or to estimate the proportion or rate of entanglement. The age class of captured and disentangled NFS is estimated using standard evaluation protocols based on size, vibrissae color, and pelage characteristics (Scheffer 1962, Vladimirov & Nikulin 1993). To minimize disturbance, we move slowly with a low profile, stop if animals show signs of stress, and wear clothing that blends into the surroundings.
Group round-up surveys randomly sample males on the hauling ground and entangled and non-entangled NFS have an equal probability of detection. Crew samples the non-pup males by encircling the inland hauling ground they occupy and herding NFS further inland. Crew subsequently releases the NFS single-file towards the water, counts every individual and notes those entangled to determine the proportion of NFS entangled. Additionally, the counts can be further sub-divided into harvestable (approximately 2-4 years old) and non-harvestable (greater than 5 years old) animals to facilitate comparison with historical entanglement data. All entangled and non-entangled NFS in a particular age or sex category observed in the count are included in the sample. During the release of the rounded-up NFS, those entangled NFS can be captured and disentangled. The round-up method allows for a higher probability of detecting entangled NFS and therefore disentangling them. The method is more technically complicated in that it requires a greater number of people involved and results in greater incidental harassment of NFS not entangled. Due to the higher rate of detection of entangled NFS and the higher probability of successful capture the round-up method significantly improves the survival of a greater number of entangled NFS and provides a more accurate estimate of entanglement for the minor negative effect of additional incidental harassment of non-pup males. In addition, we will capture, restrain, weigh, and record the number of individually tagged males detected during group round-up surveys (as described in Projects 2 and 4).
Entangled NFS will be captured with either a noose pole or capture net as described in Antonelis (1992). Animals will be restrained on a specially designed restraint board or within the capture net (as described in Project 2). Because our methods to detect entangled NFS will result in the detection of flipper-tagged NFS we will use the same methods to capture flipper-tagged NFS to weigh (as described in Project 2) and read their tag number. Captures begin by slowly crawling towards the subject while making maximal use of the local topography (e.g. boulders) to allow close approach without being detected. Such an approach minimizes disturbance to any surrounding NFS while getting within capture range of the potential subjects. While securely restrained, the NFS is disentangled (e.g. the net, packing band, etc. is cut with side cutting pliers and carefully removed). Disentanglements typically take between 5-20 minutes per animal. We attempt to clearly mark seals that are captured and disentangled by shearing away a visible line of guard hair on the neck/shoulder region of each side to expose the light-colored under-fur or apply All-flex narrow tags to the trailing edge of the front flippers (as described in Project 2). This is done to ensure that the entangled portion of the population is sampled "with replacement" (Fowler 1987). Marking previously entangled NFS will allow us to individually compare the survival of known disentangled NFS to similar aged NFS not entangled that are part of the tagged cohort in the MML vital rates program. NFS are released after all crew members, but one, have moved 5-10 m away and remain low while the neoprene harness is removed, and the restraint headboard is lifted, allowing the NFS to move away. By conducting captures in the manner described above, disturbance to adjacent NFS will be minimized. In situations where capture attempts will cause unacceptable disturbance to rookery or haulout areas, sightings will be recorded, photographed and the location of entangled NFS(s) will be monitored to the greatest extent possible to increase the likelihood of future capture opportunities. An estimate of the number of NFS disturbed during any capture attempt will be recorded.