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Exposure and resulting tissue concentrations of various elements from natural and anthropogenic sources are influenced by multiple factors, such as geographic location, age, diet, and metabolic rate, that can influence wildlife health. Essential and non-essential elements were assessed in lanugo and whole blood collected in 2019 from 102 Steller sea lion (Eumetopias jubatus) pups from two rookeries from the western and central Aleutian Islands: Agattu (WAI, n = 54) and Ulak Islands (CAI, n = 48). Rookery, sex, dorsal standard length, and trophic ecology (ẟ 15 N, ẟ 13 C values) effects on element concentration were evaluated. Significant differences in element concentrations of lanugo were exhibited across rookeries (p < 0.05), except for zinc (Zn). For example, higher mercury (Hg) and selenium (Se) concentrations were observed in WAI than CAI, while other elements were lower in WAI. Whole blood showed higher sulfur (S) and Se concentrations in CAI compared to WAI, while WAI had elevated strontium (Sr) and Hg concentrations relative to CAI. Trophic ecology significantly influenced most element concentrations, possibly due to regional variations in adult female feeding and food web dynamics. Interactions between elements were found in lanugo across both rookeries, with varying strengths. Whole blood displayed less pronounced yet consistent associations, with variable intensities. Essential elements sodium (Na), potassium (K), and calcium (Ca) formed a distinct group whose interaction is crucial for nervous system function and muscle contraction. Another group comprised zinc (Zn), iron (Fe), manganese (Mn), magnesium (Mg), phosphorous (P), S, and Se, which are known for indirectly interacting with enzyme function and metabolic pathways. Hg and Se formed a distinct group probably due to their known chemical interactions and physiological protective interactions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Mercury, a toxic metal released by various human activities, exerts environmental stress through its bioaccumulation and biomagnification, particularly in marine habitats. South American fur seals (Arctocephalus australis) and sea lions (Otaria flavescens) reproduce on the Atlantic coast of Uruguay. As top predators, they can accumulate toxic levels of mercury and are often used as sentinel species for monitoring ecosystem health. Fur seals prey on pelagic species, such as fish and squid, while sea lions consume coastal-benthic prey. We analyzed the total mercury concentration (THg) in hair and the trophic habits (δ 13 C and δ 15 N) of females from both species. The average THg concentration in adult female sea lions (30.5 ± 9.3 μg/g dry weight) was significantly higher than in fur seals (6.3 ± 2.5 μg/g dry weight). Additionally, the mean δ 15 N and δ 13 C values were significantly higher in sea lion (δ 15 N: 19.2 ± 0.6‰, δ 13 C: -13.8 ± 0.2‰) compared to fur seals (δ 15 N: 16.5 ± 0.5‰, δ 13 C: -15.5 ± 0.6‰). Our results suggest that different trophic levels and feedings areas affect the THg concentration in Uruguayan Otariids. Notably, at the intraspecific level, the THg concentration in sea lions increased with δ 13 C values, suggesting a link to coastal feeding habits. This indicates that coastal feeding behaviors, compared to feeding in pelagic environments, enhance mercury bioaccumulation in Otariids along the Uruguayan coast, with the discharge of freshwater from the Río de la Plata (one of the largest estuaries in South America) basin identified as a potential mercury source. THg concentrations found in female sea lion hair are the highest reported in Otariids globally. Mercury levels exceeded toxic thresholds observed in other mammals and could pose significant health risks. Our findings may explain why sea lions were particularly affected by the avian influenza outbreak in Uruguay compared to fur seals. Monitoring the declining sea lion population is crucial, making our results significant for integrated conservation and management strategies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Mercury is a global contaminant that bioaccumulates in a tissue-specific manner in long-lived predators such as Steller sea lions (SSL). Bone is a well-preserved material amenable for studying millennial scale trends; however, little is known about the distribution and variability of total mercury concentrations ([THg]) within individual bones and among bone elements in SSL. We assessed SSL bone [THg] variability with respect to physiologic age, bone type, longitudinally within a bone, and among bone elements. Pup bones (mean ± SD; 31.4 ± 13.58 ppb) had greater [THg] than adults (7.9 ± 1.91 ppb). There were greater and more variable [THg] within individual long bones near epiphyses compared to mid-diaphysis. Pup spongy bone in ribs (62.7 ± 44.79 ppb) had greater [THg] than long bones (23.5 ± 8.83 ppb) and phalanges (19.6 ± 10.78 ppb). These differences are likely due to variability in bone composition, growth, and turnover rate. This study informs standardized sampling procedures for [THg] in bone to improve interpretations of mercury variability over time and space., Competing Interests: Declaration of competing interest Julie Avery reports financial support was provided by the National Science Foundation. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Total mercury concentrations ([THg]) exceed thresholds of concern in some Steller sea lion (Eumetopias jubatus) tissues from certain portions of the Aleutian Islands, Alaska. We applied compound-specific stable isotope analyses of both carbon and nitrogen in amino acids from fish muscle tissue to quantify the proportional contributions of primary production sources and trophic positions of eight prey species (n = 474 total) that are part of Steller sea lion diets. Previous THg analyses of fish muscle, coupled with monomethylmercury analyses of a subset of samples, substantiated previous findings that fishes from the west of Amchitka Pass, a discrete oceanographic boundary of the Aleutian Archipelago, have higher muscle Hg concentrations relative to fishes from the east. The δ 13 C values of essential amino acids (EAAs) in fish muscle demonstrated that although most fishes obtained their EAAs primarily from algae, some species varied in the extent to which they relied on this EAA source. The δ 15 N values of phenylalanine (0.9 to 7.8 ‰), an indicator of the isotopic baseline of a food web, varied widely within and among fish species. Trophic position estimates, accounting for this baseline variation, were higher from the west relative to the east of the pass for some fish species. Trophic magnification slopes using baseline-corrected trophic position estimates indicated similar rates of Hg biomagnification to the east and west of Amchitka Pass. Multiple linear regression models revealed that trophic position was the most important driver of fish muscle [THg] with less variation explained by other parameters. Thus, higher trophic positions but not the rate of Hg biomagnification to the west of Amchitka Pass may play a role in the regional differences in both fish and Steller sea lion [THg]. Although, differences in Hg contamination and uptake at the base of the east and west food webs could not be excluded., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Similar to the several pinniped and a few terrestrial carnivore species, the Steller sea lion has a seasonal synchronized mating scheme enabled by a female reproductive cycle that includes embryonic diapause, delayed implantation, and pseudopregnancy (a state in which the corpus luteum produces progesterone for approximately as long as in pregnant females). Due to this, circulating systemic progesterone concentrations cannot be used to differentiate pregnant and nonpregnant females during early gestation. With the use of advanced measurement technologies such as liquid chromatography tandem mass spectrometry (LC-MS/MS) additional steroid hormones are measurable which can provide additional information on the endocrine pathways throughout gestation. Our objectives were to further characterize endocrine patterns in female Steller sea lion pregnancy by 1) quantifying longitudinal profiles of hormone metabolites in pregnant and non-pregnant female sera, and 2) evaluating hormone profiles to identify pregnant animals within the early stage of gestation. Three gestation stages were delineated based on what is believed to be the period of implantation (September-October): EARLY (August- November), MID (December-February), and LATE (March to May). Five steroids, Progesterone (P 4 ), 5α-dihydroprogesterone (DHP), 17αOH-progesterone (17OHP), 20αOH-progesterone (20OHP), and androstenedione (A 4 ), were detected in both pregnant and non-pregnant animals. A significant difference in P 4 concentrations was measured between EARLY and MID gestation (p ≤ 0.01) in both pregnant and non-pregnant animals. During MID gestation there was a significant difference (p ≤ 0.05) between pregnant and non-pregnant animals in all pregnanes measured. Significant patterns of correlation between P 4 and 17OHP and between P 4 and DHP were detected during EARLY and MID gestation in non-pregnant animals. While those significant correlations also exist in EARLY pregnant animals, this pattern was lost by MID gestation. This loss of correlation suggests a potential shift in progesterone metabolism from ovarian to alternative tissue (e.g. fetal gonads or adrenal glands) by MID gestation in Steller sea lions. We were unable to identifying a steroid hormone biomarker capable of differentiating pseudopregnancy from pregnant animals and conclude that such a biomarker likely falls outside of the traditional progesterone metabolic pathway., (Published by Elsevier Inc.)

The objective of this study was to establish a single-dose pharmacokinetic profile for orally administered itraconazole in California sea lions (Zalophus californianus). Twenty healthy rehabilitated juvenile California sea lions were included in this study. Itraconazole capsules were administered orally with food at a target dose of 5-10 mg/kg. Blood samples were collected from each animal at 0 hr and at two of the following timepoints: 0.5, 1, 2, 4, 6, 8, 12, 24, 48, and 72 hr. Quantitative analysis of itraconazole in plasma samples was performed by high-performance liquid chromatography. An average maximum concentration of 0.22 µg/ml ± 0.11 was detected 4 hr after administration. The average concentration fell to 0.12 µg/ml ± 0.11 by 6 hr and 0.02 µg/ml ± 0.02 at 12 hr. At no point did concentrations reach 0.5 µg/ml, the concentration commonly accepted for therapeutic efficacy. While this formulation was well tolerated by the sea lions, oral absorption was poor and highly variable among individuals. These data indicate that a single oral dose of itraconazole given as a capsule at 5-10 mg/kg, under the conditions used in this study, does not achieve therapeutic plasma concentrations in California sea lions., (© 2020 John Wiley & Sons Ltd.)

Contaminant exposure is particularly important for species and populations of conservation concern, such as the Steller sea lion (Eumetopias jubatus). We used blubber samples (n = 120) to determine organochlorine concentrations, including polychlorinated biphenyls (PCBs) and dichloro-diphenyl-trichloroethane (DDTs), and blood samples (n = 107) to estimate total body lipids based on the hydrogen isotope dilution method. We assessed the influence of age, sex, condition, and geographic area on contaminant concentrations in blubber and contaminant body load. The concentration of ΣPCBs was highest in pups (<6 months) from the Aleutian Islands, and the concentrations in males were higher than females in all regions. The ΣPCBs and ΣDDTs concentrations and loads decreased with increasing mass in pups, however, there were no regional or sex differences in contaminant load. Within each of the five age classes, the concentrations of ΣPCBs and ΣDDTs decreased with increasing mass, but overall these OCs increased with age. Further, accounting for the lipid content, a potential proxy for energy balance, in the animal load reduced or removed the regional and sex effects present in age models for contaminants. We propose, that adjusting OCs concentration by the lipid content of the blubber sample alone may not fully account for the variability in OC concentrations associated with differences in condition or energy states between young Steller sea lions., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Vernix caseosa, the white waxy coating found on newborn human skin, is thought to be a uniquely human substance. Its signature characteristic is exceptional richness in saturated branched chain fatty acids (BCFA) and squalene. Vernix particles sloughed from the skin suspended in amniotic fluid are swallowed by the human fetus, depositing BCFA/squalene throughout the gastrointestinal (GI) tract, thereby establishing a unique microbial niche that influences development of nascent microbiota. Here we show that late-term California sea lion (Zalophus californianus) fetuses have true vernix caseosa, delivering BCFA and squalene to the fetal GI tract thereby recapitulating the human fetal gut microbial niche. These are the first data demonstrating the production of true vernix caseosa in a species other than Homo sapiens. Its presence in a marine mammal supports the hypothesis of an aquatic habituation period in the evolution of modern humans.

Vitamin D plays a central role in calcium homeostasis of most vertebrates, and is obtained in different species through diet, dermal synthesis, or a combination of both. The aim of this study was to determine the predominant routes of Vitamin D synthesis in three disparate species, brown kiwi (Apteryx mantelli), tuatara (Sphenodon punctatus), and New Zealand sea lions (Phocarctos hookeri). We surveyed plasma concentrations of 25-hydroxyvitamin D 2 and D 3 , analysed environmental conditions and life history factors, and determined the ability of skin samples to synthesise Vitamin D 3 on exposure to ultraviolet-B radiation. There was variation in the plasma/serum 25-hydroxyvitamin D 3 concentrations between and within the species studied, with wild kiwi having the lowest concentrations and NZ sea lions the highest. Kiwi skin produced small but measurable amounts of Vitamin D 3 , while tuatara skin produced Vitamin D 3 concentrations higher than that of kiwi. New Zealand sea lion skin produced the highest amount of Vitamin D 3 and differed from the other two species in this study in that Vitamin D 3 was present in skin before UV-B exposure. The results from this study show that all three species studied retained the ability to use both dietary and dermal sources of Vitamin D, although there was interspecies variation in the magnitude of dermal synthesis. Comparisons between these species show that there are differences in their Vitamin D pathways, but suggest that there are more factors contributing to these pathways than might be expected solely from life history characteristics.

Chronic stress can have detrimental effects on an individual's health and reproductive success. The use of cortisol quantification as an indicator of stress in free-ranging cetaceans and phocids is increasing but no studies have applied this technique on blubber in otariids. We measured cortisol concentrations in blubber samples obtained from California sea lions, Zalophus californianus, stranded in San Diego County and those incidentally killed in the California drift gillnet fishery. We also measured progesterone concentrations to assess female reproductive status and, in males, as a potential secondary measure of adrenal steroid production. Blubber cortisol and progesterone values were compared across demographic groups (sex and maturity), season, and proportion blubber lipid extracted. Stranded animals (247.3±70.767 SE ng/gblubber) had significantly higher cortisol concentrations compared to fishery bycaught (8.1±2.108 SE ng/gblubber) animals. These findings are likely driven by inherent differences in the cause of death and associated nutritional state coupled with the mean duration of expiration for these two groups of animals (i.e., the duration from an animal's initial perception of the threat-to-self until death). The duration of transition from healthy state to death in stranded animals is on the order of many hours to weeks while in fishery bycaught animals, this transition occurs much more rapidly (i.e., seconds to tens of minutes). The presumed longer duration of the mortality event in stranded animals gives sufficient time for elevated cortisol to diffuse into the blubber. No significant differences between demographic groups, or season were found. However, blubber cortisol declined inversely with proportion blubber lipid extracted, suggesting utility in assessing long-term nutritional status. Blubber progesterone was significantly higher in mature females than immature females (153.8±54.546 SE ng/gblubber and 9.7±3.60 SE ng/gblubber respectively), containing on average 15 times more progesterone, irrespective of pregnancy state. Additionally, a significant relationship between mean cortisol and progesterone was found in males with >35% blubber lipid (p<0.0001). This study is an initial step in validating blubber cortisol and progesterone concentrations as a potential marker of stress response and reproductive state, respectively, in otariids. Especially when paired with dart biopsying, this approach could represent a relatively rapid way to assess baseline stress, nutritional status and reproductive states in otariids while minimizing the effects of sampling., (Copyright © 2017. Published by Elsevier Inc.)

Individuals within populations often differ substantially in habitat use, the ecological consequences of which can be far reaching. Stable isotope analysis provides a convenient and often cost effective means of indirectly assessing the habitat use of individuals that can yield valuable insights into the spatiotemporal distribution of foraging specialisations within a population. Here we use the stable isotope ratios of southern sea lion (Otaria flavescens) pup vibrissae at the Falkland Islands, in the South Atlantic, as a proxy for adult female habitat use during gestation. A previous study found that adult females from one breeding colony (Big Shag Island) foraged in two discrete habitats, inshore (coastal) or offshore (outer Patagonian Shelf). However, as this species breeds at over 70 sites around the Falkland Islands, it is unclear if this pattern is representative of the Falkland Islands as a whole. In order to characterize habitat use, we therefore assayed carbon (δ13C) and nitrogen (δ15N) ratios from 65 southern sea lion pup vibrissae, sampled across 19 breeding colonies at the Falkland Islands. Model-based clustering of pup isotope ratios identified three distinct clusters, representing adult females that foraged inshore, offshore, and a cluster best described as intermediate. A significant difference was found in the use of inshore and offshore habitats between West and East Falkland and between the two colonies with the largest sample sizes, both of which are located in East Falkland. However, habitat use was unrelated to the proximity of breeding colonies to the Patagonian Shelf, a region associated with enhanced biological productivity. Our study thus points towards other factors, such as local oceanography and its influence on resource distribution, playing a prominent role in inshore and offshore habitat use.

Cefovecin is a third-generation cephalosporin developed as an aqueous solution for use by the subcutaneous route in dogs and cats. This study evaluated the duration of cefovecin plasma concentrations after single intramuscular (IM) or subcutaneous (SC) injection at different doses in 10 Patagonian sea lions (Otaria flavescens). Blood samples were collected serially from the day of the injection up to 60-90 days post-injection. Plasma drug concentrations were determined by high performance liquid chromatography-UV detection and pharmacokinetic parameters were calculated by non-compartmental analysis. No reactions or side effects associated with the drug were observed in any of the studied animals. Both routes showed very similar pharmacokinetic behaviour. Elimination half-life (11.3-21.6 days, SC; 13.1-15.9 days, IM) and mean residence time (17.6-36.8 days SC; 16.5-25.4 days IM) were, in all cases and doses, considerably longer than those previously reported for any other species. Based on these findings, and preliminary data on specific pathogen sensitivity, cefovecin was found to be a very promising antimicrobial for Patagonian sea lions, in particular those that are difficult to access or that are under certain rehabilitation conditions., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The relationships of selected organochlorine (OC) contaminants between blubber, blood, feces, and milk of young, free-ranging Steller sea lions (Eumetopias jubatus) were examined. Both between and within each tissue there was considerable individual variation. In spite of the variation, similar patterns were observed across the tissues for most of the selected PCB congeners. In all four tissues, the major PCB congeners were PCB101, PCB118, PCB138, and PCB153. The most prominent congener, both as a weight (ng/g lipid) and as a percentage of summed PCBs (∑PCBs), was PCB 153. Comparisons between paired tissues showed that ∑DDTs in blubber samples were related to concentrations in blood, feces, and milk. The ∑PCBs in blubber were related to concentrations in milk and fecal samples, though the relationship with feces was weak. Our findings show milk samples, in particular, are useful for assessing OCs in young sea lions. Blubber concentrations of PCB101, PCB118, and PCB138 were an order of magnitude higher than those in milk, supporting the biomagnification of these PCB congeners in SSL tissues. The findings indicate alternative tissues may be used as indicators of relative contaminant exposure in lieu of surgical blubber biopsy., (Copyright © 2015 Elsevier B.V. All rights reserved.)

We measured total mercury (THg) concentrations in California sea lions (Zalophus californianus) and examined how concentrations varied with age class, colony, and sex. Because Hg exposure is primarily via diet, we used nitrogen (δ (15)N) and carbon (δ (13)C) stable isotopes to determine if intraspecific differences in THg concentrations could be explained by feeding ecology. Blood and hair were collected from 21 adult females and 57 juveniles from three colonies in central and southern California (San Nicolas, San Miguel, and Año Nuevo Islands). Total Hg concentrations ranged from 0.01 to 0.31 μg g(-1) wet weight (ww) in blood and 0.74 to 21.00 μg g(-1) dry weight (dw) in hair. Adult females had greater mean THg concentrations than juveniles in blood (0.15 vs. 0.03 μg(-1) ww) and hair (10.10 vs. 3.25 μg(-1) dw). Age class differences in THg concentrations did not appear to be driven by trophic level or habitat type because there were no differences in δ (15)N or δ (13)C values between adults and juveniles. Total Hg concentrations in adult females were 54 % (blood) and 24 % (hair) greater in females from San Miguel than females from San Nicolas Island, which may have been because sea lions from the two islands foraged in different areas. For juveniles, we detected some differences in THg concentrations with colony and sex, although these were likely due to sampling effects and not ecological differences. Overall, THg concentrations in California sea lions were within the range documented for other marine mammals and were generally below toxicity benchmarks for fish-eating wildlife.

Proteomic studies including marine mammals are rare, largely due to the lack of fully sequenced genomes. This has hampered the application of these techniques toward biomarker discovery efforts for monitoring of health and disease in these animals. We conducted a pilot label-free LC-MS/MS study to profile and compare the cerebrospinal fluid from California sea lions with domoic acid toxicosis (DAT) and without DAT. Across 11 samples, a total of 206 proteins were identified (FDR<0.1) using a composite mammalian database. Several peptide identifications were validated using stable isotope labeled peptides. Comparison of spectral counts revealed seven proteins that were elevated in the cerebrospinal fluid from sea lions with DAT: complement C3, complement factor B, dickkopf-3, malate dehydrogenase 1, neuron cell adhesion molecule 1, gelsolin, and neuronal cell adhesion molecule. Immunoblot analysis found reelin to be depressed in the cerebrospinal fluid from California sea lions with DAT. Mice administered domoic acid also had lower hippocampal reelin protein levels suggesting that domoic acid depresses reelin similar to kainic acid. In summary, proteomic analysis of cerebrospinal fluid in marine mammals is a useful tool to characterize the underlying molecular pathology of neurodegenerative disease. All MS data have been deposited in the ProteomeXchange with identifier PXD002105 (http://proteomecentral.proteomexchange.org/dataset/PXD002105)., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

There are various interspecies differences in xenobiotic-metabolizing enzymes. It is known that cats show slow glucuronidation of drugs such as acetaminophen and strong side effects due to the UGT1A6 pseudogene. Recently, the UGT1A6 pseudogene was found in the Northern elephant seal and Otariidae was suggested to be UGT1A6-deficient. From the results of measurements of uridine diphosphate-glucuronosyltransferase (UGT) activity using liver microsomes, the Steller sea lion, Northern fur seal, and Caspian seal showed UGT activity toward 1-hydroxypyrene and acetaminophen as low as in cats, which was significantly lower than in rat and dog. Furthermore, UGT1A6 pseudogenes were found in Steller sea lion and Northern fur seal, and all Otariidae species were suggested to have the UGT1A6 pseudogene. The UGT1 family genes appear to have undergone birth-and-death evolution based on a phylogenetic and synteny analysis of the UGT1 family in mammals including Carnivora. UGT1A2-1A5 and UGT1A7-1A10 are paralogous genes to UGT1A1 and UGTA6, respectively, and their numbers were lower in cat, ferret and Pacific walrus than in human, rat, and dog. Felidae and Pinnipedia, which are less exposed to natural xenobiotics such as plant-derived toxins due to their carnivorous diet, have experienced fewer gene duplications of xenobiotic-metabolizing UGT genes, and even possess UGT1A6 pseudogenes. Artificial environmental pollutants and drugs conjugated by UGT are increasing dramatically, and their elimination to the environment can be of great consequence to cat and Pinnipedia species, whose low xenobiotic glucuronidation capacity makes them highly sensitive to these compounds., (© The Author 2015. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

Marine mammals are repeatedly exposed to elevated extra-thoracic pressure and alveolar collapse during diving and readily experience alveolar expansion upon inhalation - a unique capability as compared to terrestrial mammals. How marine mammal lungs overcome the challenges of frequent alveolar collapse and recruitment remains unknown. Recent studies indicate that pinniped lung surfactant has more anti-adhesive components compared to terrestrial mammals, which would aid in alveolar opening. However, pulmonary surfactant composition has not yet been investigated in odontocetes, whose physiology and diving behavior differ from pinnipeds. The aim of this study was to investigate the phosphatidylcholine (PC) composition of lung surfactants from various marine mammals and compare these to a terrestrial mammal. We found an increase in anti-adhesive PC species in harp seal (Pagophilus groenlandicus) and California sea lion (Zalophus californianus) compared to dog (Canus lupus familiaris), as well as an increase in the fluidizing PCs 16:0/14:0 and 16:0/16:1 in pinnipeds compared to odontocetes. The harbor porpoise (a representative of the odontocetes) did not have higher levels of fluidizing PCs compared to dog. Our preliminary results support previous findings that pinnipeds may have adapted unique surfactant compositions that allow them to dive at high pressures for extended periods without adverse effects. Future studies will need to investigate the differences in other surfactant components to fully assess the surfactant composition in odontocetes., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Senile plaques (SPs) and cerebral amyloid angiopathy (CAA) consisting of β-amyloid (Aβ) are major features in the brain of Alzheimer's disease (AD) patients and elderly humans and animals. In this study, we report the finding of SPs and CAA in an aged sea lion (30 years), which is the first demonstration of AD-related pathological changes in a marine animal. Histologically, SPs were observed at the cerebral cortex, most frequently at the frontal lobe, with two morphologically different types: the small round type and the large granular type. Only the small round SPs were positive for Congo red staining. The SPs were equally immunoreactive to Aβ40 and Aβ42 and were mainly composed of Aβ with an N-terminal pyroglutamate residue at position 3. Amyloid depositions at vessel walls were noted at the meninges and within the parenchyma. Interestingly, double immunofluorescence staining for Aβ40 and Aβ42 showed that the two subtypes were deposited segmentally in different parts of the vessel walls. The lesions observed in the sea lion suggest that Aβ deposition is widely present in various animal species, including marine mammals; however, the peculiar deposits similar to cotton wool plaques and the specific pattern of CAA are characteristic features of this animal.

Concentrations of total mercury ([THg]) and selenium ([TSe]) were measured in several tissue compartments in Steller sea lion (Eumetopias jubatus) pups; in addition we determined specific compartment and body burdens of THg. Compartmental and body burdens were calculated by multiplying specific compartment fresh weight by the [THg] (summing compartment burdens equals body burden). In all 6 pup tissue sets (1) highest [THg] was in hair, (2) lowest [THg] was in bone, and (3) pelt, muscle and liver burdens contributed the top three highest percentages of THg body burden. In 5 of 6 pups the Se:Hg molar ratios among compartments ranged from 0.9 to 43.0. The pup with the highest hair [THg] had Se:Hg molar ratios in 9 of 14 compartments that were ⩽ 0.7 potentially indicating an inadequate [TSe] relative to [THg]., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

California sea lions (Zalophus californianus) are abundant human-sized carnivores with large gyrencephalic brains. They develop epilepsy after experiencing status epilepticus when naturally exposed to domoic acid. We tested whether sea lions previously exposed to DA (chronic DA sea lions) display hippocampal neuropathology similar to that of human patients with temporal lobe epilepsy. Hippocampi were obtained from control and chronic DA sea lions. Stereology was used to estimate numbers of Nissl-stained neurons per hippocampus in the granule cell layer, hilus, and pyramidal cell layer of CA3, CA2, and CA1 subfields. Adjacent sections were processed for somatostatin immunoreactivity or Timm-stained, and the extent of mossy fiber sprouting was measured stereologically. Chronic DA sea lions displayed hippocampal neuron loss in patterns and extents similar but not identical to those reported previously for human patients with temporal lobe epilepsy. Similar to human patients, hippocampal sclerosis in sea lions was unilateral in 79% of cases, mossy fiber sprouting was a common neuropathological abnormality, and somatostatin-immunoreactive axons were exuberant in the dentate gyrus despite loss of immunopositive hilar neurons. Thus, hippocampal neuropathology of chronic DA sea lions is similar to that of human patients with temporal lobe epilepsy., (Copyright © 2013 Wiley Periodicals, Inc.)

The concentrations of PCB congeners in the blubber and liver of mother Steller sea lions (Eumetopias jubatus; SSLs) and their fetuses from the coast of Hokkaido, Japan in 2008, 2010 and 2012 were analyzed by HRGC-HRMS, in order to elucidate PCB congener profiles and maternal-to-fetal transfer of PCBs in SSLs. ΣPCBs in the fetuses were 1400 ± 660 (the mean ± SD) ng/g-fat in the blubber and 570 ± 320 ng/g-fat in the liver, respectively. There was a concern that SSLs had been contaminated by PCBs during the fetal period. The concentrations of the major congeners in the blubber and liver were a correlation between the fetus and mother (blubber: r=0.9934, liver: r=0.9160; P ≦ 0.05). The results indicated that PCBs in the fetuses came from the mothers. #177 and #199 showed no correlations between the fetus and the mother in the blubber and liver. This indicated a selective capture by some natural protector such as the placenta., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Steller sea lion (Eumetopias jubatus) populations have undergone precipitous declines through their western Alaskan range over the last four decades with the leading hypothesis to explain this decline centering around changing prey quality, quantity, or availability for this species (i.e., nutritional stress hypothesis). Under chronic conditions of reduced food intake sea lions would conserve energy by limiting energy expenditures through lowering of metabolic rate known as metabolic depression. To examine the potential for nutritional stress, resting metabolic rate (RMR) and body composition were measured in free-ranging juvenile Steller sea lions (N = 91) at three distinct geographical locations (Southeast Alaska, Prince William Sound, Central Aleutian Islands) using open-flow respirometry and deuterium isotope dilution, respectively. Average sea lion RMR ranged from 6.7 to 36.2 MJ d(-1) and was influenced by body mass, total body lipid, and to a lesser extent, ambient air temperature and age. Sea lion pups captured in the Aleutian Islands (region of decline) had significantly greater body mass and total body lipid stores when compared to pups from Prince William Sound (region of decline) and Southeast Alaska (stable region). Along with evidence of robust body condition in Aleutian Island pups, no definitive differences were detected in RMR between sea lions sampled between eastern and western populations that could not be accounted for by higher percent total body lipid content, suggesting that that at the time of this study, Steller sea lions were not experiencing metabolic depression in the locations studied.

Total mercury concentrations ([THg]) measured in western Aleutian Island Steller sea lion pup hair were the highest maximum [THg] documented in this endangered species to date. Some pups exceeded concentrations at which other fish-eating mammals can exhibit adverse neurological and reproductive effects (21% and 15% pups above 20 and 30 μg/g in hair, respectively). Of particular concern is fetal exposure to mercury during a particularly vulnerable stage of neurological development in late gestation. Hair and blood [THg] were highly correlated and 20% of pups sampled in the western Aleutian Islands of Alaska exceeded mammalian risk thresholds established for each of these tissues. Higher nitrogen isotope ratios suggested that pups accumulated the highest [THg] when their dams fed on higher trophic level prey during late gestation., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The concentrations of PCB congeners in the blubber and liver of male and female Steller sea lions (Eumetopias jubatus; SSLs) collected from the Shakotan Peninsula and the Nemuro Strait, Hokkaido, Japan in 2008 and 2010 were measured by HRGC-HRMS, in order to express the concentration profiles in SSLs at these regions. #153 predominated in both organs, followed by #138, #99 and #118. In males, #28 and #31 were concentrated particularly in the blubber, while #177 and #199 were accumulated specifically in the liver. The differences in these concentration profiles might be expressed by the differences in the organs and their functions. The concentrations of #99, #118, #138, #153, and #180 in the blubber appeared to correspond significantly with the differences between genders. The details of the surveys on PCB congeners in SSLs could clarify the differences in the residue levels of individual congeners for organs and genders., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Body condition indices have been useful in assessing the health of domestic and free ranging populations of terrestrial mammals. Given the high energy demand and rapid growth during the postnatal period of Steller sea lion (Eumetopias jubatus) (SSL) pups, body condition was expected to be related to concentrations of a suite of hormones (cortisol, aldosterone, thyroxine, triiodothyronine, leptin) previously associated with changes in body mass and composition in developing pinnipeds. Blood samples were collected from free ranging SSL pups of known ages and sex. A body condition index (BCI) previously developed for SSL pups based on a mass and length relationship was applied to 61 SSL pups ranging in age from 5 to 38days old. BCI was not related to pup age. Overall, male pups were larger than females and older pups were larger than younger pups. Aldosterone was negatively correlated with BCI only in female pups, whereas no relationship was observed between aldosterone and BCI in males. Further, male pups had higher aldosterone concentrations than females. Concentrations of cortisol, total and free thyroxine (T4), and total triiodothyronine (T3) decreased when regressed against the elapsed time between researchers' arrival on the rookery and removal of pup from the holding corral for blood collection. While the overall variation attributed to the rookery disturbance was low (r(2)<0.293), it may be of significance for future studies on free ranging pinnipeds. This study adds to the current knowledge of the postnatal changes in free ranging SSL pups by providing ranges of the BCI and several hormone concentrations from an apparently stable population., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Fatty acids (FAs) from blubber are often analysed to assess the diet of marine mammals. However, distribution of blubber FAs is not necessarily uniform along the body. It is therefore important to understand the deposition of dietary fat to be able to estimate the diet. We analysed the FA compositions of the thoracic ventral (T region) blubber of 28 New Zealand (NZ) sea lions Phocarctos hookeri by-caught by the southern arrow squid Nototodarus sloani fishery. Each blubber sample was divided into an inner and an outer layer. For 16 of these 28 animals, the pelvic dorsal (P) region was also sampled. The influence of body region and layer was statistically tested on the distribution of blubber FAs. We found minimal differences between the P and T regions (3 out of 29 FAs). The outer blubber layer was more concentrated in short-chain monounsaturated FAs, and less concentrated in saturated FAs, but the degree of stratification was small. Diet predictions from quantitative FA signature analysis (QFASA) applied on different body regions were similar. When applied to different blubber layers, QFASA gave some variation in the contribution of rattails (~25 % in outer blubber vs. ~12 % in inner blubber). Nonetheless, diet predicted from both layers was dominated by similar prey species: octopus, hoki and rattails. Hoki and rattails shared a similar ecological niche. Therefore, feeding ecology of NZ sea lions inferred from the inner or the outer blubber would lead to the same conclusions. In the case of NZ sea lions, the outer layer of blubber, if the only sample accessible, could be a useful tissue for diet inference from FAs.

This study provides an immunohistochemical approach to the expression of leptin in the gastrointestinal tract of the monogastric South American sea lion (Otaria flavescens), and the poligastric bottlenose dolphin (Tursiops truncatus). The specific organization of the gastrointestinal tract is examined in relation to the neuroendocrine regulation of the gut exerted by leptin. In the South American sea lion some leptin-like-immunoreactive (ir) cells, and endocrine type cells, were found in the pit of gastric mucosal folds and in the epithelium of duodenum as well as numerous neurons were detected in the submucosal and myenteric plexuses of the stomach. In the bottlenose dolphin, many leptin-like-ir cells, and exocrine type cells, were identified in the mucosal layer of the main stomach as well as several neurons and nervous fibers were detected in nervous plexuses of main stomach, pyloric stomach, proximal, and middle intestine. Our data suggest that the distribution of leptin-like peptides is similar in the two species, notwithstanding the different anatomical organization of the gastrointestinal apparatus of South American sea lion and bottlenose dolphin. These findings "suggest" the presence of a basal plan in the regulation of food intake, body weight, energy balance and of the gastrointestinal functions in general also in marine mammals with different and specific feeding habits., (Copyright © 2012 Wiley Periodicals, Inc.)

Survival depends on an animal's ability to find and acquire prey. In diving vertebrates, this ability is directly related to their physiological capability (e.g. oxygen stores). We studied the seasonal variation in oxygen stores, body temperature and body condition in California sea lions (Zalophus californianus) (CSL) as a function of seasonal variation in temperature, primary productivity, diving behavior and reproductive stage. During summer, blood oxygen stores were significantly greater and muscle oxygen stores were significantly lower than in winter. Total oxygen stores, body condition and body temperature did not change between seasons but variations in body temperature were greater during summer. Changes in oxygen stores are partly attributed to diving behavior, temperature and pregnancy that could increase oxygen consumption. Blood and muscle oxygen stores appear to be influenced by reproductive state. Blood oxygen stores are more likely influenced by diving behavior and temperature than muscle oxygen stores., (Copyright © 2012 Elsevier Inc. All rights reserved.)

One of the chemical breakdown products of nonylphenol ethoxylates, 4-nonylphenol (4-NP), accumulates in organisms and is of concern as an environmental pollutant due to its endocrine disrupting effects. We measured 4-NP levels in the seawater, sediment, and twelve organisms within the California estuary, Morro Bay, and examined biomagnification of 4-NP using stable isotope abundances (δ(15)N and δ(13)C) to quantify trophic position. 4-NP concentrations in organisms from Morro Bay included 25000±8600 ng g(-1) lw in liver of California sea lion, 14000±5600 ng g(-1) lw in liver of harbor porpoise, 138000±55000 ng g(-1) lw in liver of sea otters, 15700±3600 ng g(-1) lw in liver of seabirds, 36100±6100 ng g(-1) lw in arrow goby fish, 62800±28400 ng g(-1) lw in oysters, and 12700±1300 ng g(-1) lw in mussels. 4-NP levels generally showed a pattern of trophic dilution among organisms in Morro Bay, with exceptions of biomagnification observed between three trophic links: mussel to sea otter (BMF 10.9), oyster to sea otter (BMF 2.2), and arrow goby to staghorn sculpin (BMF 2.7). Our examination of other west coast estuaries of USA and Canada revealed that mean 4-NP concentrations in gobies and mussels from Morro Bay were significantly higher than those from a more urbanized estuary, San Francisco Bay (goby: 11100±3800 ng g(-1) lw) and from a remote estuary, Bamfield Inlet, Canada (goby: 9000±900 ng g(-1) lw, mussel: 6100±700 ng g(-1) lw). Relative to other estuaries worldwide, 4-NP levels in seawater (0.42±0.16 μg L(-1)) and sediment (53±14 ng g(-1) dw) of Morro Bay are low, but gobies and oysters have higher 4-NP levels than comparable fauna., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

We characterize for the first time the presence of DDT and its metabolites in tropical Galapagos sea lions (Zalophus wolleabeki). ∑DDT concentrations in Galapagos sea lion pups sampled in 2005 and 2008 ranged from 16 to 3070 μg/kg lipid. Concentrations of ∑DDT in pups in 2008 averaged 525 μg/kg lipid and were 1.9 times higher than that (281 μg/kg lipid) detected in pups in 2005. These concentrations are lower than those reported in many pinnipeds elsewhere, comparable to those in Hawaiian monk seals, and higher than those in southern elephant seals. The health risk characterization showed that 1% of the male pups exceeded the p,p'-DDE toxic effect concentration associated with anti-androgenic effects reported in rats. The findings provide preliminary guidance on the relationship between DDT use and ecological impacts, serving as a reference point against which possible future impact of tropical DDT use can be assessed., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The ability to use heart rate (fh) to predict oxygen consumption rates ([Formula: see text]) in Steller sea lions and other pinnipeds has been investigated in fasting animals. However, it is unknown whether established fh:[Formula: see text] relationships hold under more complex physiological situations, such as when animals are feeding or digesting. We assessed whether fh could accurately predict [Formula: see text] in trained Steller sea lions while fasting and after being fed. Using linear mixed-effects models, we derived unique equations to describe the fh:[Formula: see text] relationship for fasted sea lions resting on land and in water. Feeding did not significantly change the fh:[Formula: see text] relationship on land. However, Steller sea lions in water displayed a different fh:[Formula: see text] relationship after consuming a 4-kg meal compared with the fasting condition. Incorporating comparable published fh:[Formula: see text] data from Steller sea lions showed a distinct effect of feeding after a 6-kg meal. Ultimately, our study illustrated that both feeding and physical environment are statistically relevant when deriving [Formula: see text] from telemetered fh, but that only environment affects the practical ability to predict metabolism from fh. Updating current bioenergetic models with data gathered using these predictive fh:[Formula: see text] equations will yield more accurate estimates of metabolic rates of free-ranging Steller sea lions under a variety of physiological, behavioral, and environmental states.

The western stock of the Steller sea lion (Eumetopias jubatus) in the northern Pacific Ocean has declined by approximately 80% over the past 30 years. This led to the listing of this sea lion population as an endangered species in 1997. Chemical pollution is [corrected] one of several contributing causes. In the present study, 145 individual PCBs were determined in tissues of male sea lions from Tatitlek (Prince William Sound) and St. Paul Island (Bering Sea), and placentae from the Aleutian Islands. PCBs 90/101, 118, and 153 were abundant in all the samples. The mean toxic equivalents (TEQ) were 2.6, 4.7 and 7.4 pg/g lw in the kidney, liver, and blubber samples, respectively. The mean TEQ in placentae was 8 pg/g lw. Total PCBs concentrations (2.6-7.9 μg/g lw) in livers of some males were within a range known to cause physiological effects, further [corrected] suggesting the possibility of adverse effects on this stock., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Blubber from stranded South American sea lions (Otaria flavescens) was sampled between 1991 and 2005 on the Peninsula Valdés in Argentina and analyzed for organochlorine (OC) pollutants. Mean blubber concentrations, expressed on an extractable basis, were 686 (SD = 1,060) ng g(-1) for dichlorodiphenyl trichloroethane (tDDT) and 735 (SD = 787) ng g(-1) for polychlorinated byphenils (PCB). The OC levels were well below those associated with adverse sublethal effects and lethality in mammals. OC concentrations showed statistically significant associations with age that were positive in males and negative in females. These trends are consistent with the majority of marine mammal populations studied. There were no trends in the levels of tDDT or PCB over time. In spite of the low levels detected, OC contamination was present consistently over the 14-year period, suggesting continuous inputs from geographic redistribution.

In this study we directly compared soluble and particulate chromate cytotoxicity and genotoxicity in human (Homo sapiens) and sea lion (Eumetopias jubatus) lung fibroblasts. Our results show that hexavalent chromium induces increased cell death and chromosome damage in both human and sea lion cells with increasing intracellular chromium ion levels. The data further indicate that both sodium chromate and lead chromate are less cytotoxic and genotoxic to sea lion cells than human cells, based on an administered dose. Differences in chromium ion uptake explained some but not all of the reduced amounts of sodium chromate-induced cell death. By contrast, uptake differences could explain the differences in sodium chromate-induced chromosome damage and particulate chromate-induced toxicity. Altogether they indicate that while hexavalent chromium induces similar toxic effects in sea lion and human cells, there are different mechanisms underlying the toxic outcomes., (Copyright 2010 Elsevier Inc. All rights reserved.)

Variation in concentrations of thyroid hormones shed in feces may help to identify physiological states of animals, but the efficacy of the technique needs to be validated for each species. We determined whether a known physiological alteration to thyroid hormone production was reflected in hormone concentrations in the feces of Steller sea lions (Eumetopias jubatus). We quantified variation of triiodothyronine (T3) and thyroxine (T4) concentrations in feces following two intramuscular injections of thyrotropin (thyroid-stimulating hormone, TSH) at 24h intervals in four captive female sea lions. We found fecal T3 concentrations increased 18-57% over concentrations measured in the baseline sample collected closest to the time of the first TSH injection (p=0.03) and 1-75% over the mean baseline concentration (p=0.12) for each animal of all samples collected prior to injections. Peak T3 concentrations were greater than the upper bound of the baseline 95% confidence interval for three animals. The peak T3 response occurred 48h post-injection in three animals and 71h in the fourth. Post-injection T4 concentrations did not differ between the baseline sample collected closest to the time of the first TSH injection (p=0.29) or the mean baseline concentration (p=0.23) for each animal. These results indicate that induced physiological alterations to circulating thyroid hormone concentrations can be adequately detected through analyses of fecal T3 concentrations and that the technique may provide a means of non-invasively detecting metabolic changes in Steller sea lions., (Copyright 2009 Elsevier Inc. All rights reserved.)

Concentrations of polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs) were measured in muscle-blubber biopsy samples from 21 Galapagos sea lion (Zalophus wollebaeki) pups that were live captured in the Galapagos Islands (Ecuador) using gas chromatography/high-resolution mass spectrometry. Only traces of PBDEs were detected in one male pup, whereas PCDDs and PCDFs were not detected in any sample. The total concentration of PCBs (ΣPCB) in the pups averaged 104 μg/kg lipid (range, 49-384 μg/kg). No statistically significant differences in ΣPCB were observed among the four study sites in the Galapagos Islands. Concentrations of PCB congeners in Galapagos sea lion pups were dominated by low-molecular-weight congeners. These results suggest that global transport is the main source for PCBs in Galapagos sea lions. The ΣPCB levels were below immunotoxic and endocrine-disruption thresholds in pinnipeds, suggesting a limited risk of adverse health effects. The present study indicates that Galapagos sea lions can serve as a useful sentinel of pollutants with a long-range transport capacity and that Galapagos Islands are not exempt from the threats of global pollutants despite its remote locale.

Physiological responses to changes in energy balance are tightly regulated by the endocrine system through glucocorticoids, IGF-I and thyroid hormones. Changes in these hormones were studied in eight captive female Steller sea lions that experienced changes in food intake, body mass, body composition, and blood metabolites during summer and winter. During a period of energy restriction, one group of sea lions was fed reduced amounts of Pacific herring and another was fed an isocaloric diet of walleye pollock, after which both groups returned to their pre-experimental diets of herring. Cortisol was negatively and IGF-I was positively associated with changes in body mass during periods of energy restriction (mass loss associated with increase in cortisol and decrease in IGF-I) and refeeding (body mass maintenance associated with stable hormone concentrations in summer and compensatory growth linked to decrease in cortisol and increase in IGF-I in winter). Cortisol and IGF-I were also correlated with changes in lipid and lean mass, respectively. Consequently, these two hormones likely make adequate biomarkers for nutritional stress in sea lions, and when combined provide indication of the energetic strategy (lipid vs lean mass catabolism) animals adopt to cope with changes in nutrient intake. Unlike type of diet fed to the sea lions, age of the animals also impacted hormonal responses, with younger animals showing more intense hormonal changes to nutritional stress. Thyroid hormones, however, were not linked to any physiological changes observed in this study.

We report concentrations of several classes of organochlorines (OCs) in the blubber of California sea lions (Zalophus californianus) from the Gulf of California. Summed OC levels measured in 34 wild-ranging animals were, in general, lower than those previously reported in sea lions from the eastern Pacific. The rank order of OCs was SigmaDDTs (mean=3400 ng g(-1 ) lipid weight [lw]) > SigmaPCBs (1400 ng g(-1 ) lw) > SigmaHCHs (50 ng g(-1 ) lw) >or= SigmaCHLORs (46 ng g(-1 ) lw). The most abundant OC measured was the DDT metabolite, p,p'-DDE. No significant differences in OC profiles were found between genders or rookeries. Although the mean concentrations of OCs measured in adult males and females were similar, only adult females had significantly higher (p<0.05) mean blubber concentrations of summation SigmaDDTs and summation SigmaHCHs than pups.

The levels of six trace metals (Hg, Se, Pb, Cu, Fe, and Zn) were measured in the hair of California sea lion pups from eight rookeries in the Gulf of California, with the aim of relating these levels with the animals feeding habits, trophic level, and delta(15)N signatures at each locality. Trace metal levels were seen to vary significantly between sample sites as were tendencies associated with feeding habits. Mercury concentrations showed a clear positive correlation with trophic level (r = 0.73, p = 0.06) as well as the delta(15)N values of the rookeries (r = 0.79, p = 0.033). This was explainable via the process of biomagnification, although other elements did not follow the same tendency. High Cu concentrations in the hair (18 and 47 microg g(-1); ANOVA, p < 0.05) were observed at San Pedro Mártir and San Esteban, where the consumption of squid is higher than at other localities (approximately 17% and approximately 27% of the diet, respectively). Se and Hg concentrations correlated positively (r = 0.73, p = 0.038), probably the result of detoxification processes, a phenomenon previously reported in marine mammals. In light of the high degree of philopatry among sea lions in the Gulf and their pups' dependence exclusively upon their mother's milk during the first months of life, the analysis of trace metals in the easily sampled hair of the pups, offers an indirect way of monitoring the toxicology of the sea lion populations.

The metabolic costs of foraging and the management of O2 and CO2 stores during breath-hold diving was investigated in three female Steller sea lions (Eumetopias jubatus) trained to dive between 10 and 50 m (N=1142 dives). Each trial consisted of two to eight dives separated by surface intervals that were determined by the sea lion (spontaneous trials) or by the researcher (conditioned trials). During conditioned trials, surface intervals were long enough for O2 to return to pre-dive levels between each dive. The metabolic cost of each dive event (dive+surface interval; DMR) was measured using flow-through respirometry. The respiratory exchange ratio (VO2/VCO2) was significantly lower during spontaneous trials compared with conditioned trials. DMR was significantly higher during spontaneous trials and decreased exponentially with dive duration. A similar decrease in DMR was not as evident during conditioned trials. DMR could not be accurately estimated from the surface interval (SI) following individual dives that had short SIs (<50 s), but could be estimated on a dive by dive basis for longer SIs (>50 s). DMR decreased by 15%, but did not differ significantly from surface metabolic rates (MRS) when dive duration increased from 1 to 7 min. Overall, these data suggest that DMR is almost the same as MRS, and that Steller sea lions incur an O2 debt during spontaneous diving that is not repaid until the end of the dive bout. This has important consequences in differentiating between the actual and 'apparent' metabolic rate during diving, and may explain some of the differences in metabolic rates reported in pinniped species.

Patterns and contents of major acidic glycosphingolipids in the kidney of three marine mammalian species, the Steller sea lion (Pinnipedia), the rough-toothed dolphin and the broad-beaked dolphin (Odontoceti), were examined, and compared with those of terrestrial mesic mammals. The profile of major acidic glycosphingolipids was not significantly different between the terrestrial and marine mammals: predominant gangliosides were GM3 and GD3, and major sulfoglycolipids were SM4s and SM3. On the other hand, the total concentration (nmol/g wet tissue) of sulfoglycolipids was considerably higher in the marine mammals (2.3-3.0 times) than that in the terrestrial mesic mammals with comparable body weights. In contrast, there was no significant difference in the level of renal glycolipids-bound sialic acid between the marine and the terrestrial mammals. These results suggest that higher expression of renal sulfoglycolipids in marine mammals may contribute to the maintenance of osmotic balance of their body fluid against sea water.

The endangered Western population of the Steller sea lion declined for three decades for uncertain reasons. We present baseline data of metal concentrations in pups as a first step towards investigating the potential threat of developmental exposures to contaminants. Seven metals were investigated: arsenic, cadmium, silver, aluminum, mercury, lead and vanadium. Vanadium was detected in only a single blubber sample. Mercury appears to be the most toxicologically significant metal with concentrations in the liver well above the current action level for mercury in fish. The concentrations of aluminum, arsenic, silver, cadmium and lead were present in one-fourth to two-thirds of all samples and were at either comparable or below concentrations previously reported. Neither gender nor region had a significant effect on metal burdens. Future work should consider metal concentrations in juveniles and adults and toxicological studies need to be performed to begin to assess the toxicity of these metals.

Given that many marine mammals display seasonal energetic priorities, it is important to investigate whether the impact of unexpected food restriction differs during the year. Steller sea lions (Eumetopias jubatus) fed restricted diets for up to 9 days during spring, summer, fall, and winter lost an average of 10% of their initial body mass. We tracked changes in the levels of three hormones (cortisol, total thyroxine-TT4, total triiodothyronine-TT3) and one blood metabolite (blood urea nitrogen-BUN) following a food restriction in relation to season, body mass, body composition, and metabolism. Degree of changes in cortisol, TT3, and BUN after food restriction was significantly affected by season. The greatest changes in cortisol (+231%), BUN (+11.4%), TT4 (-23.3%), and TT3 (-35.6%) occurred in the winter (November/December) when rates of body mass loss were also greatest. Changes in cortisol levels were positively related to total body mass loss, while changes in TT3 levels were negatively related. While greater increases in BUN were related to greater rates of mass loss, the use of BUN levels as an indicator of metabolic state is complicated by the type and level of food intake. The observed changes in hormone levels support morphological data suggesting Steller sea lions may be more strongly impacted by short-term, reduced energy intake during winter than at other times of the year.

In domestic animal medicine, changes in serum enzyme levels are routinely used as diagnostic tools to detect liver disease. Hepatic disease occurs in pinnipeds, but limited data are available on the tissue distribution of serum enzymes in marine mammals. The objectives of this study were to determine the tissue distribution of seven serum enzymes in three pinniped species. Enzymes evaluated were alanine aminotransferase (ALT), aspartate aminotransferase (AST), sorbitol dehydrogenase (SDH), lactate dehydrogenase (LDH), creatine kinase (CK), alkaline phosphatase (ALP), and gamma-glutamyl transferase (GGT) in tissues from California sea lions (Zalophus californianus) (n = 5), harbor seals (Phoca vitulina) (n = 5), and northern elephant seals (Mirounga angustirostris) (n = 5) that stranded and then died at a rehabilitation center. Samples were evaluated in duplicate from liver, skeletal muscle, cardiac muscle, kidney, adrenal, spleen, pancreas, lung, lymph node, and intestine. Patterns of tissue enzyme distribution were similar in all species, with SDH activity highest in liver and kidney, CK activity highest in skeletal and cardiac muscle, ALP activity highest in adrenal, and GGT activity highest in the kidney. Aspartate aminotransferase and LDH activities were less specific, with high activity in multiple tissues. Tissue ALT activity was high in the liver of all species, but was also high in cardiac muscle (California sea lions), skeletal muscle (harbor seals), and kidney (elephant seals). These results suggest that concurrent analysis of SDH, ALT, and CK would provide high specificity and sensitivity for the detection of hepatic lesions, and allow differentiation of liver from skeletal muscle lesions in pinniped species.

Eight free-ranging juvenile Steller sea lions (SSL; 6 males, 2 females; 14-20 months) temporarily held under ambient conditions at the Alaska SeaLife Center were physiologically challenged through exogenous administration of adrenocorticotropic hormone (ACTH). Four individuals (3 males, 1 female) underwent ACTH challenge in each of two seasons, summer and winter. Following ACTH injection serial blood and fecal samples were collected for up to 3 and 96 h, respectively. A radioimmunoassay (RIA) was validated for leptin, and using a previously validated RIA for cortisol, collected sera were analyzed for both cortisol and leptin. ACTH injection resulted in a 2.9-fold increase (P=0.164) in leptin which preceded a 3.2-fold increase (P=0.0290) in cortisol by 105 min in summer. In winter, a 1.7-fold increase in leptin (P=0.020) preceded a 2.1-fold increase (P=0.001) in serum cortisol by 45 min. Mean fecal corticosteroid maxima were 10.4 and 16.7-fold above baseline 28 and 12 h post-injection and returned to baseline 52 and 32 h post-injection, in summer and winter, respectively. Data indicate acute activity in juvenile adrenal glands is detectable in feces approximately 12-24 h post-stimulus in either season, with a duration of approximately 40 h in summer and 20 h in winter. Changes in serum cortisol proved statistically significant both seasons and elevated concentrations were detected by 30 min post-stimulus (baseline 64.8+/-4.2; peak 209.5+/-18.3 ng/ml: summer; baseline 87.0+/-15.7; peak 237.6+/-10.0 ng/ml: winter), whereas the changes that occurred in serum leptin proved to be significant only in winter (baseline 6.4+/-0.6; peak 18.7+/-7.0 ng/ml: summer; baseline 4.2+/-0.5; peak 7.5+/-0.6 ng/ml: winter). Changes in fecal corticosteroids proved significant only in summer (baseline 117.8+/-36.7; peak 1219.3+/-298.4 ng/g, P=0.038: summer; baseline 71.8+/-13.7; peak 1198.6+/-369.9 ng/g, P=0.053: winter) due to a high degree of individual variability in winter months. The data indicate that ACTH stimulates leptin production earlier than cortisol in both summer and winter, and that while the leptin response appears most variable in summer, fecal corticosteroids are most variable in winter.

Otariids, like other wild mammals, contend with a wide variety of energetic demands across seasons. However, due to the cryptic behaviors of this marine group, few studies have been able to examine longitudinal energetic costs or the potential impact of these costs on seasonal or annual prey requirements. Here we evaluated the changes in energy demand and intake of female California sea lions (Zalophus californianus) during reproductive (n=2 sea lions) and nonreproductive (n=3) periods. Monthly measurements included resting metabolic rate, blood hormone levels, body condition (blubber thickness and body mass), and caloric intake for adult sea lions throughout molting, late pregnancy, lactation, and postweaning. We found that maintenance energy demands decreased from 32.0 to 23.1 MJ d(-1) before pupping, remaining stable at 19.4+/-0.6 MJ d(-1) during lactation and postweaning. Energy intake rates to meet these demands showed marked changes with activity level and the reproductive cycle, reaching a peak intake of 3.6 times baseline levels during lactation. Translating this into prey demands, we find that 20,000 reproductively active females on San Nicolas Island rookeries would maximally require 4,950 metric tons of Pacific whiting during a month of the breeding season. This localized impact is reduced significantly with postbreeding dispersal and demonstrates the importance of considering spatial and temporal factors driving the energetic requirements of predators when designing marine protected areas.

Concentrations of nine heavy metals (As, Cd, Cu, Fe, Hg, Pb, Mn, Mo and Zn) were determined in the hepatic and renal tissues of 80 stranded California sea lions (Zalophus californianus). Significant age-dependant increases were observed in liver and kidney concentrations of cadmium and mercury, and renal zinc concentrations. Hepatic iron concentrations were significantly higher in females than males. Animals with suspected domoic acid associated pathological findings had significantly higher concentrations of liver and kidney cadmium; and significantly higher liver mercury concentrations when compared to animals classified with infectious disease or traumatic mortality. Significantly higher hepatic burdens of molybdenum and zinc were found in animals that died from infectious diseases. This is the largest study of tissue heavy metal concentrations in California sea lions to date. These data demonstrate how passive monitoring of stranded animals can provide insight into environmental impacts on marine mammals.

Tooth wear in marine mammals has been attributed to age, feeding habits, behavior, and contaminants. Advanced tooth wear in some California sea lions, including some of very young age (<5 yr), in the Gulf of California, suggests that there are variations in chemical composition of tooth parts, wherein the concentrations of certain trace minerals might be anomalous, making them more susceptible to erosion. The concentrations of the essential minerals Ca, P, K, Na, Fe, Mg, and Zn in the dentition of Zalophus c. californianus are documented for the first time and are compared for sea lion teeth with different degrees of wear. Canine teeth and molars from 45 skulls collected at 15 localities since 1978 were digested in perchloric acid and analyzed using atomic adsorption spectrometry, the results being expressed in milligrams per 100 g. An index of tooth wear (Id) was established, involving the average wear on the teeth and the age of the organism. No significant difference was detected in the variables, but there was one between ages (p = 0.02). A higher degree of wear was observed up to 7 yr of age than from this age onward. Mineral concentrations did not explain the excessive wear observed (correlation, p > 0.09; ANOVA, p > 0.15); however, the Ca concentration of the teeth was inversely proportional to the age of the animal (sexes combined, p = 0.026) and particularly significant for the females (r2 = 0.112, r = -0.335, p = 0.039). Females could be more prone to decalcification because of their annual bone investment in their offspring. Animals of both sexes were susceptible to tooth wear as their age increased, but the higher frequency of animals between 4 and 7 yr suggests an impact on survival at early stages probably linked to deficient feeding and chronic malnutrition.

Chronically heightened adrenal activity indexed by fecal corticosteroids has been shown to be a valid descriptor of stress in many species. As part of an ongoing investigation of adrenal activity in Steller sea lions (SSL), adrenocorticotropic hormone (ACTH) challenges were performed during the summer at the Alaska SeaLife Center (ASLC). These results were compared to earlier data from winter months. Additionally, adrenal response of free-ranging females SSL to a presumed in situ stressor, pup predation by killer whales (Orcinus orca), was evaluated as a field trial of developed methodologies. Summer ACTH results indicated that gender-dependent differences in baseline fecal corticosterone concentrations exist, with summer baseline fecal corticosterone concentrations higher in males than in females, based on season when compared with previously reported values in winter ACTH trials for this species. ACTH trials in the male in the summer resulted in 2468 ng/g basal to 10,937 ng/g maximal fecal corticosterone concentrations, a fourfold change. Female 1 exhibited a 30.5-fold increase 24 h post-ACTH stimulation (27.9-852.0 ng/g dry weight), with a return to just above baseline concentrations by hour 25. Additionally, female 2 exhibited a 64.4-fold increase at 25 h post-stimulation (31.7-2042.0 ng/g dry weight), with a return to just above baseline concentrations by hour 45. In situ female fecal corticosterone tripled 24h subsequent to orca predation on pups (54.6+/-18.5 ng/g mean pre-attack, 542.5+/-252.7 ng/g mean post-attack) during the field trial. Data from both the summer ACTH and subsequent field trial underscore the necessity of gender determination of collected scat source and knowledge of conditions at collection sites for proper interpretation of fecal corticosterone data, particularly in studies that focus on population-wide stress. With elimination of gender bias and delineation of acute response to natural stimuli, corticosterone data can now be evaluated within context and provide meaningful information about stress and potentially reproductive physiology in free-ranging Steller sea lions.

The purpose of this study was to investigate thyroid hormone concentrations, thyroxine (T4) and triiodothyronine (T3), in order to determine basal levels in Steller sea lions of different ages and over seasons. Serum concentrations of total T4 were highest in Steller sea lions followed by total T3 concentrations. Concentrations of free T4 and free T3 were three to four orders of magnitude lower. Concentrations for all four thyroid hormone measurements tended to a lower level as animals matured beyond the neonatal stage. When thyroid hormones from captive sea lions were evaluated across seasons, all thyroid hormones were highest in the July to September period. When compared across the geographic range, animals in southeast Alaska tended to have lower thyroid hormone levels, while the Steller sea lions west of Prince William Sound and animals from the Russian Far East had significantly higher concentrations. Significant inter-annual differences in concentrations were also observed across the geographic range. With an understanding of the basic changes in thyroid hormone concentrations, changes in plane of nutrition or life history states (i.e. fasting, pregnancy or lactation) can now be evaluated for their effect on the overall health of this endangered species.