Total mercury concentrations in Steller sea lion bone: Variability among locations and elements.

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.

Preliminary comparisons between a point-of-care ketometer and reference method using Steller sea lion pup whole blood and plasma.

We evaluated the Precision Xtra™ ketometer as part of a larger study categorizing fasting status of free-ranging Steller sea lion (Eumetopias jubatus; SSL) pups which necessitated the identification of plasma ß-hydroxybutyrate concentrations ([ß-HBA]) around a threshold of <0.3 and ≥0.3 mmol/l. Whole blood samples mixed with sodium heparin (NaHep) or ethylenediaminetetraacetic acid liquid anticoagulants were tested <10 minutes after collection (n = 14; triplicate technical replicates). Plasma (stored at -80°C, NaHep, Thaw1) measured via our laboratory's Reference Assay (Sigma Aldrich, St. Louis, MO, Kit #MAK041) served as the standard [ß-HBA] for ketometer comparisons. Our observed ß-HBA range (0.0-1.6 mmol/l), consistent with published [ß-HBA] of free-ranging Otariid pups, represented the lower 20% of the ketometer's range (0.0-8.0 mmol/l). The maximal coefficient of variation (%CV) of ketometer technical replicates was 9.1% (NaHep, whole blood). The majority of ketometer technical replicate sets (84%, including all matrices, anticoagulants and thawings) were identical (CV = 0%). We found linear relationships and agreement of ketometer [ß-HBA] between whole blood preserved with different anticoagulants and between whole blood and plasma (Thaw1) measurements. The ketometer produced results with linearity to the Reference Assay for both whole blood and plasma (Thaw1). We identified a non-linear relationship between plasma at Thaw1 and Thaw2 (tested four months apart, NaHep), as only samples with higher SSL [ß-HBA] decreased in concentration, and all others remained the same. With respect to categorizing SSL pup fasting in our larger study, the ketometer's %Accuracy, %Sensitivity and %Specificity for samples with Reference Assay ß-HBA <0.2 and >0.4 mmol/l were 100%. We adopted a modified procedure: plasma samples with mean ketometer concentrations ±0.1 mmol/l of 0.3 mmol/l ß-HBA were re-evaluated using the Reference Assay, improving measurement precision from tenths (ketometer) to thousandths (assay) mmol/l. The Precision Xtra™ ketometer was valuable to our application over the range of [ß-HBA] observed in SSL pup plasma and whole blood samples.

Fasting durations of Steller sea lion pups vary among subpopulations-evidence from two plasma metabolites.

Geographic differences in population growth trends are well-documented in Steller sea lions (Eumetopias jubatus), a species of North Pacific pinniped listed under the U.S. Endangered Species Act in 1990 following a marked decline in population abundance that began during the 1970s. As population growth is intrinsically linked to pup production and survival, examining factors related to pup physiological condition provides useful information to management authorities regarding potential drivers of regional differences. During dam foraging trips, pups predictably transition among three fasting phases, distinguished by the changes in the predominant metabolic byproduct. We used standardized ranges of two plasma metabolites (blood urea nitrogen and ß-hydroxybutyrate) to assign pups to fasting categories (n = 1528, 1990-2016, 12 subpopulations): Recently Fed-Phase I (digestion/assimilation-expected hepatic/muscle glycogen usage), Phase II (expected lipid utilization), transitioning between Phases II-III (expected lipid utilization with increased protein reliance), or Phase III (expected protein catabolism). As anticipated, the majority of pups were classified as Recently Fed-Phase I (overall mean proportion = 0.72) and few pups as Phase III (overall mean proportion = 0.04). By further comparing pups in Short (Recently Fed-Phase II) and Long (all other pups) duration fasts, we identified three subpopulations with significantly (P < 0.03) greater proportions of pups dependent upon endogenous sources of energy for extended periods, during a life stage of somatic growth and development: the 1) central (0.27 ± 0.09) and 2) western (0.36 ± 0.13) Aleutian Island (declining population trend) and 3) southern Southeast Alaska (0.32 ± 0.06; increasing population trend) subpopulations had greater Long fast proportions than the eastern Aleutian Islands (0.10 ± 0.05; stabilized population). Due to contrasting population growth trends among these highlighted subpopulations over the past 50+ years, both density-independent and density-dependent factors likely influence the dam foraging trip duration, contributing to longer fasting durations for pups at some rookeries.

Reduction of Paraoxonase Expression Followed by Inactivation across Independent Semiaquatic Mammals Suggests Stepwise Path to Pseudogenization.

Convergent adaptation to the same environment by multiple lineages frequently involves rapid evolutionary change at the same genes, implicating these genes as important for environmental adaptation. Such adaptive molecular changes may yield either change or loss of protein function; loss of function can eliminate newly deleterious proteins or reduce energy necessary for protein production. We previously found a striking case of recurrent pseudogenization of the Paraoxonase 1 (Pon1) gene among aquatic mammal lineages-Pon1 became a pseudogene with genetic lesions, such as stop codons and frameshifts, at least four times independently in aquatic and semiaquatic mammals. Here, we assess the landscape and pace of pseudogenization by studying Pon1 sequences, expression levels, and enzymatic activity across four aquatic and semiaquatic mammal lineages: pinnipeds, cetaceans, otters, and beavers. We observe in beavers and pinnipeds an unexpected reduction in expression of Pon3, a paralog with similar expression patterns but different substrate preferences. Ultimately, in all lineages with aquatic/semiaquatic members, we find that preceding any coding-level pseudogenization events in Pon1, there is a drastic decrease in expression, followed by relaxed selection, thus allowing accumulation of disrupting mutations. The recurrent loss of Pon1 function in aquatic/semiaquatic lineages is consistent with a benefit to Pon1 functional loss in aquatic environments. Accordingly, we examine diving and dietary traits across pinniped species as potential driving forces of Pon1 functional loss. We find that loss is best associated with diving activity and likely results from changes in selective pressures associated with hypoxia and hypoxia-induced inflammation.

Total Mercury, Total Selenium, and Monomethylmercury Relationships in Multiple Age Cohorts and Tissues of Steller Sea Lions (Eumetopias jubatus).

Steller sea lion (Eumetopias jubatus) tissue mercury concentrations increasingly above thresholds of concern occur in regions of Alaska where lack of population recovery is noted. Selenium-monomethylmercury interactions may mitigate toxicosis but may also result in functional selenium deficiency, impacting essential selenium-dependent processes. Physiologically driven differences in tissue concentrations (organotropism) of total mercury ([THg]), total selenium ([TSe]), and monomethylmercury ([MeHg+ ]) confound interpretation for various age cohorts. Archived tissues from Alaska Steller sea lions (2002-2016) were used to compare [THg], [MeHg+ ], and [TSe] across age cohorts and tissue types. Liver [THg] ranged from 0.05 to 63.7 µg/g. Fetal and pup livers had significantly lower [THg] and [TSe], higher percentage MeHg+ , and greater range of molar TSe:THg than subadult and adult livers. Molar Se:MeHg+ ratios, including Se in excess of nonmethylmercury, were dependent on [MeHg+ ] in fetuses and pups. While [THg] varied significantly by muscle type (heart vs. skeletal) and anatomical location, concentrations were strongly correlated. Biomagnification and/or bioaccumulation of THg in liver of older animals confounded comparison with other tissues; however, in fetal and pup liver [THg] correlated with other tissues. In contrast, liver [MeHg+ ] correlated with other tissues across all age classes. Fetal and pup tissues, which reflect in utero exposure and are subject to limited bioaccumulation, are ideal for assessing mercury exposure related to dam diet, including intertissue comparison, and represent key cohorts of concern. Evaluating [MeHg+ ] and [TSe] in tissues from multiple age cohorts allows better intertissue comparison, providing insight into time courses, routes of exposure, and potential for adverse effects. Environ Toxicol Chem 2022;41:1477-1489. © 2022 SETAC.

Regional variations and relationships among cytokine profiles, white blood cell counts, and blood mercury concentrations in Steller sea lion (Eumetopias jubatus) pups.

The Steller sea lion (SSL) population west of 144°W longitude experienced a significant population decline. While there appears to be a stable or increasing population trend in rookeries in the Gulf of Alaska (GOA) and Southeast Alaska (SEA), some rookeries within the Aleutian Islands (AI) have failed to recover. Previous studies found regional differences in whole blood total mercury concentrations ([THg]) showing more than 20% of AI pups had [THg] above critical thresholds for increased risk of immunological effects and other adverse outcomes. Measurements of immune cell-signaling proteins can be used to evaluate the immune status of marine mammals in relation to [THg]. We compared serum cytokine and chemokine concentrations in pups among regions (AI, eastern GOA, SEA), and examined associations among cytokines, chemokines, white blood cell (WBC) counts, and [THg]. Considering liver is an important target organ for mercury and immune protein synthesis we additionally examined the relationship of [THg] with liver-related enzymes serum aspartate (AST) and alanine aminotransferase (ALT). We observed regional differences in cytokine and chemokine measurements and immune protein associations. There was a positive association between total WBC counts and [THg] in AI pups, whereas a negative association between lymphocytes and [THg] in SEA pups. These findings may indicate regional variation in proliferation and differentiation of hematopoietic cells, differences in immune system development, and/or a difference in antigenic stimuli. No associations between [THg] and cytokines, chemokines, AST or ALT were found. Observed regional differences in cytokine and chemokine milieu during gestational and early development in SSL pups could lead to an imbalance in cell differentiation that could impact immunological resiliency in juvenile and adult life stages. We report concentration ranges of a suite of cytokines and chemokines which may prove to be a useful metric for ecotoxicology and risk assessment studies in SSLs and other wildlife.

Assessment of clinical outcomes associated with mercury concentrations in harbor seal pups (Phoca vitulina richardii) in central California.

Monomethyl mercury (MeHg+) from the diet can cause mild to severe neurotoxicosis in fish-eating mammals. Chronic and low-level in utero exposure also can be neurotoxic, as documented in laboratory animal studies and epidemiologic investigations. In free-ranging animals, it is challenging to study low-level exposure related neurotoxicosis, and few studies have investigated the relationship between mercury (Hg) and adverse outcomes in wild populations. Relative to Hg concentrations on admission we evaluated different types of behaviors for 267 Pacific harbor seal (HS; Phoca vitulina richardii) pups at The Marine Mammal Center from 2015 to 2019 during rehabilitation after stranding and maternal separation. Admitted HS pups underwent a clinical exam; including sex and weight determination, and hair (partly lanugo grown in utero) and blood samples were collected for total Hg concentration ([THg]) determination. All pups were monitored weekly (behavior assessments included response to tactile stimulation, movement, swimming, interactions with other seals, hand feeding, and feeding independently), and days in rehabilitation and survival were recorded. There was a significant negative correlation between [THg] and responses to tactile stimulation and movements, measured in both hair and whole blood (p < 0.05). This relationship was found both during the intensive care unit (ICU) stage, and during the pool stage of rehabilitation. Additionally, there was a significant association between greater [THg] and number of days spent in rehabilitation, although there was no relationship between [THg] and survival. There was a significant sex difference, with greater [THg] in female pups, which contrasts with previously published findings in juvenile and adult harbor seals. Our findings support small, but significant associations between gestational THg exposure and clinical effects for tactile sensory response and movement, and longer rehabilitation durations for HS pups, although there was considerable variability among animals.

Organochlorine contaminant concentrations in blubber of young Steller sea lion (Eumetopias jubatus) are influenced by region, age, sex, and lipid stores.

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.

REGIONAL AND AGE-RELATED VARIATIONS IN HAPTOGLOBIN CONCENTRATIONS IN STELLER SEA LIONS ( EUMETOPIAS JUBATUS) FROM ALASKA, USA.

Varying concentrations of the highly conserved acute phase response protein, haptoglobin, can indicate changes to the health and disease status of mammals, including the Steller sea lion (SSL; Eumetopias jubatus). To better understand factors relating to acute phase response in SSLs, circulating haptoglobin concentrations (Hp) were quantified in plasma collected from 1,272 individuals sampled near rookeries and haulouts off the coast of Alaska, US. We compared Hp in SSLs between sexes and among different age classes (young pups, young-of-the-year, yearlings, subadults, and adults) sampled within distinct regions in Alaska (Aleutian Islands, Gulf of Alaska, Southeast Alaska). Regional and age-related differences were observed, particularly in younger SSLs. No sex-related differences were detected. We identified weakly significant relationships between Hp and hematology measurements including white blood cell counts and hematocrit in young pups from the Aleutian Islands and Southeast Alaska. No relationship between Hp and body condition was found. Lastly, a nonlinear relationship of plasma Hp and whole blood total mercury concentrations (THg) was observed in SSLs from the endangered western distinct population segment in Alaska. These results demonstrated that regional variation in Hp, especially in younger SSLs, may reflect regional differences in health and circulating THg.

Enhanced quantification of serum immunoglobulin G from a non-model wildlife species, the Steller sea lion (Eumetopias jubatus), using a protein A ELISA.

Immunoglobulins (Ig) are proteins that preserve immune homeostasis and are quantified to infer changes to the acquired humoral immune response in mammals. Measuring Ig in non-model wildlife for immune surveillance often requires ingenuity, and rigorous standardization of methodologies to provide reliable results especially when lacking species-specific reagents. We modified and optimized existing ELISA methodology utilizing the binding properties of Staphylococcus-derived Protein A (PrtA) to immunoglobulin G (IgG). We enhanced the assay for quantifying IgG in Steller sea lion (SSL) serum using critical quality control measures including dilution linearity, spike and percent recoveries, and internal controls. Of the modifications made, heat treatment of SSL serum enhanced accuracy and precision of IgG measurements by improving linearity and percent recovery in parallel dilutions and serum spikes. Purified canine IgG standard was not affected by heat inactivation. These results support that confounding serum proteins interfere with binding of PrtA with IgG demonstrating the need for heat treatment of serum to optimize IgG quantification using the PrtA-ELISA. Further, essential validation measures ensure proper assay performance. Consequently, the improved PrtA-ELISA provides species-independent IgG detection with validation criteria to enhance accuracy and precision for addressing future immunological questions in non-model wildlife in clinical, ecological, and conservation contexts.

Evaluating Hair as a Predictor of Blood Mercury: The Influence of Ontogenetic Phase and Life History in Pinnipeds.

Mercury (Hg) biomonitoring of pinnipeds increasingly utilizes nonlethally collected tissues such as hair and blood. The relationship between total Hg concentrations ([THg]) in these tissues is not well understood for marine mammals, but it can be important for interpretation of tissue concentrations with respect to ecotoxicology and biomonitoring. We examined [THg] in blood and hair in multiple age classes of four pinniped species. For each species, we used paired blood and hair samples to quantify the ability of [THg] in hair to predict [THg] in blood at the time of sampling and examined the influence of varying ontogenetic phases and life history of the sampled animals. Overall, we found that the relationship between [THg] in hair and blood was affected by factors including age class, weaning status, growth, and the time difference between hair growth and sample collection. Hair [THg] was moderately to strongly predictive of current blood [THg] for adult female Steller sea lions (Eumetopias jubatus), adult female California sea lions (Zalophus californianus), and adult harbor seals (Phoca vitulina), whereas hair [THg] was poorly predictive or not predictive (different times of year) of blood [THg] for adult northern elephant seals (Mirounga angustirostris). Within species, except for very young pups, hair [THg] was a weaker predictor of blood [THg] for prereproductive animals than for adults likely due to growth, variability in foraging behavior, and transitions between ontogenetic phases. Our results indicate that the relationship between hair [THg] and blood [THg] in pinnipeds is variable and that ontogenetic phase and life history should be considered when interpreting [THg] in these tissues.

Relating fibropapilloma tumor severity to blood parameters in green turtles Chelonia mydas.

Fibropapillomatosis is a neoplastic disease that is commonly found in the green turtles Chelonia mydas in tropical and subtropical regions of the world. In the current project, juvenile green turtles were captured with large-mesh tangle nets in the Indian River Lagoon and on nearshore reefs of Indian River County, Florida, USA, in 1998 and 1999. The purpose of the study was to determine the relationship between the severity of the disease and the general health of green turtles as indicated by blood parameters. All turtles were measured and examined, and the overall severity of the disease was rated by the size, number, and location of external fibropapilloma tumors. Hematocrit, total protein, and hemoglobin concentration were measured and compared with tumor scores (tumor severity appraisal). As the tumor score increased, the blood parameters of turtles decreased; for instance, the percentage of decrease in hematocrit for mildly afflicted, moderately afflicted, and severely afflicted groups were 2.6, 18.3, and 45.5%, respectively. Severely afflicted turtles suffered from anemia, while individuals with mild affliction did not.

Assessment of mercury and selenium tissular concentrations and total mercury body burden in 6 Steller sea lion pups from the Aleutian Islands.

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].

No evidence of metabolic depression in Western Alaskan juvenile Steller sea lions (Eumetopias jubatus).

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.

Maternal Steller sea lion diets elevate fetal mercury concentrations in an area of population decline.

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.

Serum chemistry reference ranges for Steller sea lion (Eumetopias jubatus) pups from Alaska: stock differentiation and comparisons within a North Pacific sentinel species.

Blood chemistry and hematologic reference ranges are useful for population health assessment and establishing a baseline for future comparisons in the event of ecosystem changes due to natural or anthropogenic factors. The objectives of this study were to determine if there was any population spatial structure for blood variables of Steller sea lion (Eumetopias jubatus), an established sentinel species, and to report reference ranges for appropriate populations using standardized analyses. In addition to comparing reference ranges between populations with contrasting abundance trends, data were examined for evidence of disease or nutritional stress. From 1998 to 2011, blood samples were collected from 1,231 pups captured on 37 rookeries across their Alaskan range. Reference ranges are reported separately for the western and eastern distinct population segments (DPS) of Steller sea lion after cluster analysis and discriminant function analysis (DFA) supported underlying stock structure. Variables with greater loading scores for the DFA (creatinine, total protein, calcium, albumin, cholesterol, and alkaline phosphatase) also were greater for sea lions from the endangered western DPS, supporting previous studies that indicated pup condition in the west was not compromised during the first month postpartum. Differences between population segments were likely a result of ecological, physiological, or age related differences.

Mercury concentrations in hair from neonatal and juvenile Steller Sea Lions (Eumetopias jubatus): implications based on age and region in this northern Pacific marine sentinel piscivore.

Mercury is a global contaminant of concern for the fetus and the neonate of piscivores. Methylmercury, produced within marine ecosystems, is of particular concern as a readily absorbed neurotoxicant transported across the blood brain barrier and transplacentally. In the North Pacific Ocean, Steller sea lions are broadly distributed apex predators and, as such, integrate complex food webs and the associated exposure and possible adverse effects of toxic and infectious agents. Hair, including lanugo, was examined using regional and age groupings to assess mercury concentrations in young Alaskan Steller sea lions. The highest concentrations of mercury occurred in the youngest animals, likely via in utero exposure. Based on the adverse developmental outcomes of methylmercury toxicity this specific cohort is of concern. Regionally, higher concentrations of mercury were observed in the endangered western population of Steller sea lions and mirrored patterns observed in human biomonitoring studies of Alaskan coastal communities. These data have broader implications with respect to human and ecosystem health as Steller sea lions rely on similar prey species and foraging areas as those targeted by commercial fisheries and subsistence users and are therefore valuable sentinels of marine ecosystem health.

Seasonal differences in biochemical adaptation to fasting in juvenile and subadult Steller sea lions (Eumetopias jubatus).

Nine Steller sea lions (Eumetopias jubatus) aged 1.75-6 yr were experimentally fasted for 7-14 d during the breeding and nonbreeding seasons to identify changes in plasma metabolites that are indicative of fasting and to determine whether the ability of sea lions to fast varies seasonally or with age. Although some animals approached the limit of their protein-sparing ability by the end of our fasting experiments, there was no sign of irreversible starvation biochemistry. Plasma blood urea nitrogen (BUN) concentrations decreased in all animals within the first week of fasting, reflecting a shift to a fasting-adapted state; however, significant increases in plasma BUN concentration at the end of the nonbreeding season fasts suggest that subadult Steller sea lions were not able to maintain a protein-sparing metabolism for a full 14 d during the nonbreeding season. In contrast, juveniles were able to enter protein sparing sooner during the nonbreeding season when they had slightly higher initial percent total body lipid stores than during the breeding season. Subadult and juvenile sea lions had low circulating ketone body concentrations compared with young sea lion pups, suggesting an age-related difference in how body reserves are utilized during fasting or how the resulting metabolites are circulated and catabolized. Our data suggest that metabolite concentrations from a single blood sample cannot be used to accurately predict the duration of fast; however, threshold metabolite concentrations may still be useful for assessing whether periods of fasting in the wild are unusually long compared with those normally experienced.

The effects of age, season and geographic region on thyroid hormones in Steller sea lions (Eumetopias jubatus).

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.

Ontogeny of total body oxygen stores and aerobic dive potential in Steller sea lions (Eumetopias jubatus).

Two key factors influence the diving and hence foraging ability of marine mammals: increased oxygen stores prolong aerobic metabolism and decreased metabolism slows rate of fuel consumption. In young animals, foraging ability may be physiologically limited due to low total body oxygen stores and high mass specific metabolic rates. To examine the development of dive physiology in Steller sea lions, total body oxygen stores were measured in animals from 1 to 29 months of age and used to estimate aerobic dive limit (ADL). Blood oxygen stores were determined by measuring hematocrit, hemoglobin, and plasma volume, while muscle oxygen stores were determined by measuring myoglobin concentration and total muscle mass. Around 2 years of age, juveniles attained mass specific total body oxygen stores that were similar to those of adult females; however, their estimated ADL remained less than that of adults, most likely due to their smaller size and higher mass specific metabolic rates. These findings indicate that juvenile Steller sea lion oxygen stores remain immature for more than a year, and therefore may constrain dive behavior during the transition to nutritional independence.