Thesis (M.E.S.)--The Evergreen State College, 2007. / Title from title screen (viewed 1/24/2008). "MES thesis." Includes bibliographical references (p. 79-88).

Introduction California sea lion (Zalophus californianus) health is severely compromised by domoic acid toxicosis, which occurs in high levels during harmful algal blooms of Pseudonitzschia australis along the coast of California. Current diagnostic protocols are often inconclusive due to a 2-48 hour window of detectability within the urinary, circulatory, and gastric systems (Cook, et al. 2011 and Monte, Pers Comm, 2012). Past studies suggest that Z. californianus, with domoic acid toxicosis, commonly display abnormal behaviors (Goldstein, et al. 2008). However, many of these abnormal behaviors are also associated with other diagnoses and are therefore unreliable as diagnostic indicators. This study fills in a knowledge gap relating to abnormal behavior types and their correlation to domoic acid toxicosis and helps solve the problem of current, inconclusive, diagnostic protocols. In this study, my objectives were to identify abnormal behaviors correlated to domoic acid toxicosis, create a diagnostic ethogram, determine the applicability of the method in the field, and determine the applicability of triage based on the relationship between abnormal behaviors and domoic acid levels. Methods I conducted focal animal continuous scans (continuous observation of a single animal at a time, for a set period) with continuous data entry, on animals admitted to the Marine Mammal Center (main study location during 2011-2013) and the Marine Mammal Care Center (comparison location, 2013). I conducted my observations from behind a blind to prevent both human habituation and behavioral influence of the observer. Observations lasted between 10-15 minutes (10 minutes per pen in 2011, 15 minutes per animal in 2012-2013). Subjects were selected based on an admit date no later than 7 days from the observation date. I conducted focal animal continuous scans at Pier 39, a haul out location, in the San Francisco Bay. Animals included in the study had identifying marks or were isolated from other animals (making them easy to identify). I observed animals once per observation day with a total observation period not exceeding 15 minutes per animal. I logged domoic acid levels in feces, urine, and serum (collected by veterinary staff and analyzed with liquid chromatography and bioassays for the presence of domoic acid). I then compared these results to the types and severity of abnormal behaviors displayed by the domoic acid toxicosis sample. Results Results from data collected at the Marine Mammal Center suggest that head weaving (Wilcoxon, p Results from the Pier 39 study suggest that behavioral criteria may be applicable for ruling out domoic acid toxicosis in groups of animals. However, I did not test the method during times of harmful algal blooms. Therefore, the applicability of the method for use as a diagnostic tool in the field is unknown and further research is required. Results for the triage study were inconclusive. The number of animals that tested positive for domoic acid was small and not suitable for statistical analysis. I suggest further research into triage abilities. Conclusion Based on the results of these studies, I can conclude that behavioral analysis offers a reliable diagnostic tool for rescued Z. californianus. Practitioners can use behavioral diagnostic criteria with confidence for the diagnosis of domoic acid toxicosis in Z. californianus.

Pinnipeds exhibit a wide range of lactation strategies that vary from just a few days to nearly three years in duration. Phocids have a relatively short, intense nursing period culminating with weaning after just a few days or weeks, while dependent otariids generally take several months of consuming a combined milk and solid food diet before being completely independent. The transition to nutritional independence can be particularly challenging for newly weaned pups, which must adjust to behavioral, physiological and nutritional changes as a milk diet is replaced with solid food. An interruption in energy resources during this formative stage could result in a prioritization away from growth, maintenance, or activity resulting in suboptimal development. Three groups of ex situ California sea lion (Zalophus californianus) pups were examined during the initial period of independence after they were weaned at approximately five, seven, and nine months of age. Absolute growth rates of pups were calculated and changes in body composition were estimated using blubber depth measurements and deuterium oxide dilution to determine if weaning age had an effect on subsequent pup development and growth. Blood urea nitrogen and blood glucose levels were observed for their response to changes in body condition, while thyroid hormone levels in the blood were examined as a possible nutritional stress indicator during the pup's transition to solid food. When compared to in situ pups, the 5 month old pups in the present study had significantly greater body mass (39.6 ± 1.6 kg, p < 0.01), axillary girth (85.3 ± 2.9 cm, p < 0.01), and axillary blubber depth (2.3 ± 0.1 cm, p < 0.01) compared to 5 month old in situ pups (26.6 ± 5.2 kg / 70.6 ± 5.34 cm / 1.5 ± 0.2 cm). Nine month old ex situ pups had significantly greater axillary blubber depth (3.7 ± 0.9 cm, p < 0.01) and total body lipid percentage (24.9 ± 4.7%, p= 0.01) than in situ pups (1.5 ± 0.2 cm / 17.1 ± 4.9%). Although all pups in the present study survived the transition to solid food, there were apparent differences in how the different age groups responded physiologically. The five month old pups began the switch to solid food with the lowest overall blubber depth (2.6 ± 0.9 cm) and experienced the greatest change in body mass (-8.5 ± 1.6 kg) and composition while taking the longest to begin physiological recovery (31.7 ± 1.2 d). In contrast, the 9 month old pups entered the transition with more energy reserves (24.9 ± 4.7% TBL), began consuming solid food sooner (16.3 ± 0.6 d), and were able to utilize reserves more efficiently to minimize loss and promote faster growth.

Made available in DSpace on 2014-08-20T14:31:28Z (GMT). No. of bitstreams: 1 dissertacao_eliane_machado_pereira.pdf: 4235511 bytes, checksum: 28dcd80ec8888a8603565a7809054107 (MD5) Previous issue date: 2012-03-15 / This study verified helminth parasites infection in gastrointestinal tract, lungs, heart, and kidneys of southern sea lions, Otaria flavescens, from south coast of Rio Grande do Sul State, Brazil. Twenty-nine sea-lions were found dead on the beaches, whose carcasses had mild state of decomposition, were necropsied. The organs were collected during field necropsies for laboratory analyses were 24 small and large intestines, 24 livers including parenchyma and gall bladder, 29 stomachs, 24 hearts, and 24 pairs of kidneys. The organs were maintained frozen at -20°C until their processing. A sieve with 150µm mesh was used for screening the parasites. All content retained was analyzed under stereomicroscope. The helminthes were collected, counted, fixed in AFA, stained with carmine, and clarified in beechwood creosote. The small intestines were divided into three segments that were separately analyzed to record the distribution of helminthes by preference sites. Kolmorogov-Smirnov test was used to verify the type of data distribution. Comparison of mean abundance of infection between age classes was performed through Wilcoxon test at significant level of 0.05. Correlations between infection intensity, sex, total length of the individual, and length of small and large intestines were determined using Pearson s Correlation. Action® software version 1.1 was applied for statistical analyses. Among 29 specimens of O. flavescens 23 were males, three females, and three individuals whose sex could not be determined. The average length of the animals was 2.14±0.31m (1.58 to 2.64m) including 13 sub-adults and 16 adults. A total of 996 specimens of Contracaecum ogmorhini were recorded, especially in the stomach (10.34% of prevalence), 42,145 specimens of Corynosoma australe (100% of prevalence) and 512 of Bolbosoma turbinella (50% of prevalence) were found. Two species of trematodes were found: Stephanophrora uruguayense (Prev. 4.17%) and Ascocotyle (Phagicola) longa (Prev. 33.33% ), a estimated total of 1,988.202 specimens. Cestodes were found in only one of the hosts (4.16% of prevalence) which presented four scoleces. Macroscopically, liver, gall bladder, heart, lungs, and kidneys did not contain parasites. No significant correlation was observed between infection intensity, mean abundance, sex, total length of the host, or length of intestines. Infections levels were similar between sub-adults and adults sea lions. This is the first record of Diphyllobothrium sp., Bolbosoma turbinella, Contracaecum ogmorhini, Ascocotyle (Phagicola) longa, and Stephanoprora uruguayense in O. flavescens in Brazilian waters. As regards parasite fauna of O. flavescens, our data are different from those previously reported for specimens from Pacific coast of South America. / Este estudo analisou os helmintos parasitos gastrointestinais, pulmonares, cardíacos e renais do O. flavescens no litoral sul do Rio Grande do Sul, Brasil. Foram necropsiados 29 leões-marinhos, cujas carcaças apresentavam baixo estado de decomposição. Os órgãos coletados mediante necropsia a campo para análise em laboratório foram 24 intestinos delgado e grosso, 24 fígados incluindo parênquima e vesícula biliar e 29 estômagos, 24 corações e 24 pares de rins. Os órgãos foram congelados a 20oC até o seu processamento. Para a triagem dos parasitos foi usada peneira com malha de 150 µm e todo o conteúdo retido foi analisado sob microscópio estereoscópico. Os helmintos foram fixados em AFA, corados com Carmin e clarificados com creosoto de Faia. Os intestinos delgados foram divididos em três segmentos que foram analisados separadamente para registrar a distribuição dos helmintos por sítios de preferência. Teste de Kolmorogov-Smirnov foi utilizado para verificar tipo de distribuição dos dados. A comparação da abundância média de infecção entre classes etárias foi realizada através do teste de Teste de Wilcoxon usando nível de significância de 0.05. As correlações entre intensidade de infecção, sexo, comprimento total do individuo e comprimento dos intestinos delgado e grosso foram verificadas usando a Correlação de Pearson. Para as análises estatísticas usou-se o software Action® versão 1.1. De 29 espécimes de O. flavescens 23 eram machos, três fêmeas e em três indivíduos o sexo não pode ser determinado. A média do comprimento total dos animais foi 2,14±0,31m (1,58 - 2,64m), sendo 13 subadultos e 16 adultos. Foram registrados 996 espécimes de Contracaecum ogmorhini presentes principalmente no estômago, (prevalência 10%). Registrou-se 42.145 espécimes de Corynosoma australe (Prev. 100%) e 512 de Bolbosoma turbinella (Prev. 50%). Duas espécies de trematódeos foram coletados: Stephanophrora uruguayense (Prev. 4.17% ) e Ascocotyle (Phagicola) longa (Prev. 33.33% ) totalizando 1.988.202 espécimes. Cestódeos foram encontrados em apenas um hospedeiro (Prev. 4,16%) que apresentou 4 escóleces. Macroscopicamente, fígado, vesícula biliar, coração, pulmões e rins examinados não estavam parasitados. Nenhuma correlação significativa foi observada entre a intensidade de infecção, sexo, comprimento total ou comprimento dos intestinos. Este é primeiro registro de Diphyllobothrium sp., Bolbosoma turbinella e Contracaecum ogmorhini em O. flavescens em águas brasileiras. No que se diz respeito à fauna parasitária de O. flavescens, as espécies encontradas no presente estudo não são as mesmas previamente citadas para leãomarinho-do-sul da costa pacífica da América do Sul.

Environmental fluctuations in the eastern Pacific Ocean are reflected in the tissues of some of its most vulnerable apex predators, the Peruvian fur seal (PFS) Arctocephalus australis ssp.and the South American sea lion (SASL) Otaria byronia.These large pinnipeds live in sympatry along the Pacific coastline of South America and forage within the neritic waters over the continental shelf. The coastal waters off Peru are a region of great environmental fluctuations due to periodic, oscillating El Niño- La Niña- Southern Oscillation (ENSO) events, which result in ecosystem-wide food web changes. Pinniped vibrissae (whiskers) are continuously growing keratinous tissues and reflect the incorporation of prey from the region. Fine resolution sampling along their length provided trophic information on a weekly to monthly time scale over several years. Vibrissae were obtained from 2010-2016 from female (n=47), male (n=17) and newborn pup (n=6) fur seals and male (n=6) sea lions collected during pinniped rookery health assessments. Stable isotope ratios (δ13C and δ15N) in vibrissae infer temporal primary production and dietary variations in individuals. ENSO conditions were dictated by the sea surface temperature anomaly (SSTA) recordings from the Niño 1+2 Index region over 12 years. Fluctuations in vibrissae δ15N were correlated to varying SSTA readings (p15N (p15N signatures (p13C revealed minor fluctuations ranging from -18.13 to -13.17‰over the 12-year period, δ13C did reveal a significant depletion and enrichment oscillation during specific points in time, such as the 2014-2016 El Niño event, which ranged from -15.09 to -13.83‰(p=0.040). Stable isotope signatures varied between genders, but not by species. Female fur seal stable isotope signatures were significantly more depleted in both δ13C and δ15N than males (p13C and δ15N were inversely correlated to each other from 2014 to 2016; this was during the strongest El Nino-Southern Oscillation event on record (p=0.002). As δ13Csignatures became more enriched, δ15N signatures depleted from 2014 until 2016. This suggests that when ENSO warm phase conditions occur, the environments resources change in historic foraging grounds, possibly forcing pinnipeds to travel farther distances offshoreto forage or altering what they feed on throughout these stressful conditions.This study serves to help understand of the impact of oceanographic influences on these pinnipeds.ENSO conditions influence the trophic dynamics and resultant survival adaptations in both of these vulnerable Peruvian marine mammals.

An increase in marine mammal stranding and die-off events has been observed along the California coast. The exact cause to explain for these recent events is unknown, but El Niño and harmful algal blooms are established sources for temporary decreases in marine mammal health. To determine whether El Niño could be causing and amplifying harmful algal blooms, particularly in Monterey Bay where they occur frequently, data was analyzed from the Marine Mammal Center in Sausalito, California. Data analysis focused on California sea lions (Zalophus californianus), because they have the largest data set and are directly affected by harmful blooms from domoic acid. Results indicated that El Niño events could be significantly harming California sea lions in Monterey Bay during the fall season. Because agricultural runoff is a known factor in causing harmful algal blooms and Monterey Bay experiences them consistently, online research was conducted on agricultural activity, agricultural runoff and nitrogen contamination from fertilizer in the water-sheds surrounding Monterey Bay. Nitrogen contamination from agricultural use is a prominent issue, therefore, I proposed some possible solutions, including cap and trade, a water recycling plant, and eco-certification, to minimize nitrogen’s impact on the environment and wildlife while allowing farmers to continue using nitrogen on crops.

The objectives of the present study were to: 1) describe the gross chemical milk composition of the New Zealand sea lion (NZSLs), Phocarctos hookeri, in early lactation; 2) validate an analytical method for sea lion milk composition; 3) investigate a series of temporal, individual and dietary factors that influence the milk composition of the NZSL and; 4) investigate the temporal and spatial differences in the fatty acids signatures of sea lion milk. A comprehensive literature review revealed that data on milk composition in otariid species is either missing or limited, that to be able to fully describe their milk composition extensive sampling was required and that the temporal, maternal and offspring factors that influence milk composition in pinnipeds are poorly understood. The review identified that considerable work has been conducted to infer diet via the application of fatty acids signature analysis of milk and blubber. There are many factors (i.e. metabolism, de novo synthesis and endogenous sources) that contribute to the differences in fatty acid composition between the diet and milk or blubber. Milk samples from NZSL were used to test whether a new method would give similar results as the standard methods of milk analysis. Agreement between analytical methods for milk components was assessed using different measures of statistical fitness and the results indicated that the new method was comparable to the standard methods and applicable to the milk of sea lions, pinnipeds and to ecological studies of lactation. Milk from NZSLs was collected over a period of seven years (1997, 1999 to 2003, and 2005) in early lactation to describe the composition of milk of NZSL and to test for differences between years. The results indicated that: i) the milk protein concentration was comparable to other species of pinnipeds; ii) the milk fat concentration and the milk energy content of NZSL is the lowest reported for otariids in early lactation; however iii) the milk fat concentration was significantly different between years. These results suggested that the milk composition of NZSLs was influenced by annual changes in the environment; however, there may be other unidentified factors. Month, maternal body condition, age, body weight and length, offspring sex and age, and attendance pattern were compared with milk components. The results identified that month, maternal body condition and age significantly affected milk fat concentration. These results and the fact that maternal body condition varied significantly between years and mothers nursing male pups had lower body condition and produced milk lower in energy content suggested that local food resources along with other unidentified factors have an effect on the reproductive success of NZSLs. To test whether the fatty acid signature analysis (FASA) of lipid rich tissues (milk, blubber and serum) of otariids could be used to infer diet a mixture of vegetable oil (with distinctive fatty acid signature) was fed to 24 lactating NZSL and tissue samples were collected at different time intervals. Significant increases in the concentration of specific fatty acids in serum and milk were observed with peaks within 12hrs and 24hrs respectively of ingestion. Concentrations in milk remained elevated for up to 72hrs and there were differential rates of incorporation into milk. These findings confirm the potential of FASA to infer the composition of the diet. The variation in milk fatty acid signatures from lactating NZSL from four years (1997, 2003, 2004 and 2005) of sampling were measured in order to test whether differences occurred between years. Fatty acids signatures from five potential prey species including the commercially important arrow squid were incorporated into the analysis to associate the changes in milk fatty acids with a shift in prey choice. The results indicated that milk fatty acid signatures were different in 1997 and 2003; however, it was not possible to relate these differences to the five prey species. The variability in the annual arrow squid catch data suggested that local food resources around the Auckland Islands may also be variable. In conclusion, the milk produced by the NZSL has the lowest concentration of fat and energy in early lactation reported for any otariid species. The main factors that contributed to changes in milk quality were stage of lactation, year and maternal body condition. The yearly variation in the quality of milk appears to be a result of their lactation strategy or to variable local food conditions that also affect maternal body condition. Therefore monitoring the annual milk quality may be a means to monitor the health of a pinniped population and potential management tool for pinniped species. This thesis has shown that annual changes in the diet of NZSL can be assessed with milk fatty acid signatures.

The coastal, indigenous communities around Alaska have subsisted on marine animals for generations, often focusing on large apex predators such as seals, sea lions, and whales. Three species of pinnipeds (harbor seal, Steller sea lion, northern fur seal) and the northern sea otter have all undergone significant population declines since the 1970s, some regions more than others. Archived vibrissae (whiskers) and body tissues from these four species were available from the Bering Sea and throughout the Gulf of Alaska from the 1990s and early 2000s. Tissues from these species are exceedingly difficult to obtain; thus, the archived tissues provided a finite and irreplaceable resource of data. Analysis of these archived tissues indicates which species, tissues, and gender bioaccumulate metals more readily. In this study twelve heavy metals (arsenic, cadmium, chromium, cobalt, copper, lead, manganese, mercury, nickel, selenium, vanadium, zinc) were analyzed in vibrissae from the four select species, and in body tissues from harbor seals and Steller sea lions. The samples were collected from three regions (southeastern, southcentral, and southwestern Alaska) during the 1990s through early 2000s. Significant differences of heavy metal concentrations in vibrissae were detected among elements (p2(110) = 454.81, p2(66) = 310.88, p

Muscle samples from 105 marine mammals stranded along the Oregon-Washington coasts (2002-2009) were tested for levels of total mercury by Cold Vapor Atomic Fluorescence Spectrometry. The total mercury present is predominantly in the form of highly toxic methylmercury. After normalizing muscle tissue to 75% water weight, due to variance in water content, Steller sea lions (Eumetopias jubatus) and northern elephant seals (Mirounga angustirostris) exhibited the highest mean concentrations of total mercury followed by harbor seals (Phoca vitulina), harbor porpoises (Phocoena phocoena), and California sea lions (Zalophus californianus); 0.34 ± 0.278, 0.34 ± 0.485, 0.21 ± 0.216, 0.17 ± 0.169 and 0.15 ± 0.126 mg/kg normalized weight, respectively. Mean normalized values demonstrated limited muscle methylmercury accumulation in these species in the Pacific Northwest. Normalizing muscle mercury concentrations eliminated variability from desiccation, and allowed for a clearer indication of the amount of mercury the animal accumulated before stranding. However, actual wet weight concentrations in some of the stranded carcasses were high enough to pose a risk to scavengers.