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.

Mercury maternal transfer in two placental sharks and a yolk-sac ray from Baja California Sur, Mexico.

Total mercury (THg) concentrations were measured in muscle and liver of two placental viviparous sharks, the Pacific sharpnose shark (Rhizoprionodon longurio) and the brown smooth-hound (Mustelus henlei); as well as in the muscle, liver, and yolk of the yolk-sac viviparous speckled guitarfish (Pseudobatos glaucostigmus) in Baja California Sur. The aim was to determine which factors could be involved in maternal transfer and resultant maternal and embryonic THg concentration. Higher THg concentrations were found in pregnant females compared to embryos paired tissues. THg concentrations of embryo tissues decreased with total length (TL), except for the muscle of the Pacific sharpnose shark. THg concentrations of embryo muscle was positively related to THg concentration in the muscle of pregnant females. Embryos TL, muscle THg concentration of pregnant females, percentage of THg concentration in embryos, along with the reproductive strategy are relevant factors required to improve our understanding of THg concentration in embryo tissues.

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.

Fetal mercury concentrations in central California Pacific harbor seals: Associated drivers and outcomes.

Mercury (Hg) is a well-known toxicant in wildlife and humans. High total Hg concentrations ([THg]) have been reported in central California harbor seals Phoca vitulina richardii. We evaluated the effects of presence/absence of early natal coat (lanugo), year (2012 to 2017), sex, stranding location, and trophic ecology (ẟ13C and ẟ15N values) on hair [THg] along coastal central California. Also examined were [THg] effects on growth rates of pups in rehabilitation and probability of release (e.g., successful rehabilitation). The [THg] ranged from 0.46-81.98 mg kg-1 dw, and ẟ15N and ẟ13C ranged from 13.6-21.5‰, and -17.2 to -13.0‰, respectively. Stranding location, year, and presence of lanugo coat were important factors explaining variation in [THg]. Seals from Sonoma and San Mateo County had higher [THg] than other locations. Seals with full or partial lanugo coat had lower [THg]. Seals from 2016 and 2017 had higher [THg] than those from 2015. Hair [THg] exceeded lower and upper toxicological thresholds (>20 mg kg-1 by year (5.88% to 23.53%); >30 mg kg-1 (0% to 12.31%)) with a pronounced increase from 2015 to 2016. Pups in 2017 had significantly higher odds ratio of [THg] above 20 mg kg-1 than pups of 2015, and pups in 2016 had significantly higher odds ratio than those from 2013 and 2015 (similar when using 30 mg kg-1). Pups in Sonoma County had the highest odds ratio for [THg] in lanugo above 20 mg kg-1. ẟ15N values were higher in 2015-2017, particularly relative to 2014, probably associated with the El Niño event. The [THg] was not a good predictor for probability of release and mass-specific growth rates in captivity. Further investigation of temporal trends of [THg] in harbor seals is warranted given the relatively high percentage of samples exceeding threshold values, particularly in the most recent sampling years.

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.

Mercury concentrations in Baja California Sur fish: Dietary exposure assessment.

Total mercury concentrations ([THg]) in muscle were determined in commercial finfish and elasmobranchs from Baja California Sur (BCS), Mexico to evaluate dietary Hg exposure for BCS communities, including the relationship of trophic ecology, length and mass with [THg] that might drive future consumption advice (e.g., recommend limited consumption of large fish for some species). The [THg] ranged from 0.06 to 528.02 µg kg-1 ww in finfish and 17.68-848.26 µg kg-1 ww in elasmobranchs. Relative to the consumption threshold set for predatory fish in Mexico, all species had a concentration below 1000 µg kg-1 ww. As expected, 16 (4.02%) and 75 (18.84%) individual fish were above advisory thresholds of 500 and 200 µg kg-1 ww, respectively. The hazard quotients (HQs) in most species were significantly <1.0, only banded guitarfish showed a significant median HQ > 1.0. Thus, the relative level of risk of high Hg exposure is low for most species.

Isotopic (δ15 N) relationship of pregnant females and their embryos: Comparing placental and yolk-sac viviparous elasmobranchs.

Nitrogen stable isotopes ratios (δ15 N) were determined for selected tissues (muscle, liver, blood and yolk) of pregnant females and their embryos of a placental viviparous species, the Pacific sharpnose shark (Rhizoprionodon longurio), and a yolk-sac viviparous species, the speckled guitarfish (Pseudobatos glaucostigmus). The R. longurio embryo tissues were 15 N enriched compared to the same tissues in the pregnant female, using the difference in δ15 N (Δδ15 N) between embryo and adult. Mean Δδ15 N was 2.17‰ in muscle, 4.39‰ in liver and 0.80‰ in blood. For P. glaucostigmus, embryo liver tissue was significantly 15 N enriched in comparison with liver of the pregnant female (Δδ15 N mean = 1.22‰), whereas embryo muscle was 15 N depleted relative to the muscle of the pregnant female (Δδ15 N mean = -1.22‰). Both species presented a significant positive linear relationship between Δδ15 N and embryo total length (LT ). The results indicated that embryos have different Δδ15 N depending on their reproductive strategy, tissue type analysed and embryo LT .

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.

Concentrations of organohalogens (PCBs, DDTs, PBDEs) in hunted and stranded Northern sea otters (Enhydra lutris kenyoni) in Alaska from 1992 to 2010: Links to pathology and feeding ecology.

Many organohalogen compounds (OHCs) are persistent organic pollutants (POPs) found in appreciable concentrations in marine predators. While production of some POPs has declined or ceased in recent decades, their capacity for global transport and bioaccumulation results in observations of unchanging or increasing concentrations in marine systems. Sea otters (Enhydra lutris) have been advocated as an environmental sentinel for contaminants due to their longevity, site fidelity and prey species that often overlap with human consumption. Using archived (1992-2010) samples of livers from Northern sea otters (n = 50) from Alaska we examine concentrations of chlordanes (CHLs), polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDTs), and polybrominated diphenyl ethers (PBDEs) and associated metabolites. We found some evidence for declining ΣPCBs over the two decades, however for most animals concentrations were low compared to toxicological thresholds. Six animals had relatively high concentrations of ΣPCBs (mean = 262,000 ng/g lipid weight), ΣDDTs (mean = 8,800 ng/g lw), and ΣPBDEs (mean = 4,600 ng/g lw), with four of these six animals experiencing hepatic parasitism or hepatitis. In order to assess whether differences in POP concentrations are associated with feeding ecology, we examined stable isotopes of C and N in archived muscle and whisker samples. In general, there were no significant relationships between ΣPOP concentrations and stable isotope ratios. There were small differences in stable isotope profiles in animals with high POP concentrations, although it was unclear if these differences were due to feeding ecology or disease processes. This study highlights the importance of considering feeding ecology and necropsy (health and disease status) data while conducting contaminant surveys, and confirms some previous reports of trends in OHCs in Alaska marine mammals.

Using carbon and nitrogen stable isotope modelling to assess dietary mercury exposure for pregnant women in Baja California Sur, Mexico.

INTRODUCTION: Previous studies of mercury (Hg) in pregnant women in the area of La Paz, Baja California Sur (BCS), Mexico found a proportion of individuals had concentrations of total Hg ([THg]) above some thresholds of concern set by health agencies. The [THg] were associated with fish and seafood consumption as well as other factors; although it was unclear which marine diet items could potentially be contributing to the concentrations observed. METHOD: We examined [THg] and monomethylmercury concentration ([MeHg+]) in the archived hair of 70 pregnant women from BCS as well as in diet items including fish, shellfish, and staple items (rice, beans, corn, and flour). We measured stable isotopes of carbon and nitrogen and employed a Bayesian stable isotope mixing model to investigate the proportion of fish and seafood in the isotopic profiles of archived hair samples. RESULTS: Concentrations of Hg species were low in staple foods and ranged from below detection limit to 5.71 parts per billion (ppb) wet weight. In hair, geometric mean [THg] was 658 ppb and [MeHg+] was 395 ppb, which were lower than previous reports. Percent MeHg+ was positively correlated with higher δ15N values. CONCLUSIONS: The largest carbon contributors to the diet of the study participants were corn and rice, and our analysis of fish contribution to diet varyingly agreed with the self-reported fish consumption. This report highlights the ability to discriminate potential sources of Hg from a diverse diet and the limitations of dietary recall studies.

HAIR, WHOLE BLOOD, AND BLOOD-SOAKED CELLULOSE PAPER-BASED RISK ASSESSMENT OF MERCURY CONCENTRATIONS IN STRANDED CALIFORNIA PINNIPEDS.

Mercury (Hg) poses a health risk to wildlife populations and has been documented at relatively high concentrations in many marine mammals, including wild-caught pinnipeds along the central California, US coast. We measured total Hg concentrations ([THg]) in hair and blood of live-stranded harbor seals (HS; Phoca vitulina), California sea lions (CSL; Zalophus californianus), and northern elephant seals (NES; Mirounga angustirostris) in California to quantify species, temporal, and spatial variability in [THg] and assess the relationships between [THg] measured by different methods (blood vs. filter paper) and in different matrices (blood vs. hair). We compared [THg] with toxicologic thresholds of concern to aid in identification of at-risk individuals or groups and better understand how the use of different methods and matrices affects assumed toxicologic risk. There was a wide range of [THg] in blood (<0.01-1.13 µg/g) and hair (0.45-81.98 µg/g), and NES had higher [THg] compared with HS and CSL. All three species had individuals with [THg] that exceeded the lower threshold for one or both matrices, but only HS pups had [THg] exceeding upper thresholds. Spatial differences in [THg] were detected, with higher concentrations in HS pups from areas surrounding San Francisco Bay, but differences were dependent on sampling year and matrix. The relationship between [THg] in blood and filter paper (r2=0.98) was strong, and differences had little influence on comparisons with toxicologic thresholds. Blood and hair [THg] were generally in agreement (r2=0.72), but large mismatches for a few seals underscore the importance of combined sampling in adverse effects studies where accurate assessment of Hg exposure is crucial. The wide range of [THg] in stranded HS pups that exceeded published thresholds of concern makes them a promising candidate for adverse effects studies, particularly because different matrices represent Hg exposure across key developmental stages.

Trophic Structure and Biomagnification of Total Mercury in Ray Species Within a Benthic Food Web.

Stable isotopes of C (δ13C) and N (δ15N) were used to explore the trophic structure and evaluate mercury (Hg) biomagnification in the food web of muscle of three commercially important ray species from the Pacific coast of Baja California Sur (PCBCS): the shovelnose guitarfish (Pseudobatos productus), banded guitarfish (Zapteryx exasperata), and bat ray (Myliobatis californica). The food web of these ray species predominately consisted of zooplankton, three species of fish, and five species of invertebrates. Mean δ15N values in all species ranged from 10.54 ± 0.18‰ in zooplankton to 17.84 ± 0.81‰ in the shovelnose guitarfish. Mean δ13C values ranged from - 22.05 ± 0.75‰ in the red crab to - 15.93 ± 0.78‰ in the bat ray. Mean total Hg concentration ([THg]) in all species ranged from 0.0009 ± 0.0002 mg kg-1 ww in zooplankton to 0.24 ± 0.19 mg kg-1 ww in the banded guitarfish. The food web magnification factor was 6.38 and significantly greater than 1.0. The present study describes [THg] biomagnification in the benthic food web of three ray species of the PCBCS. This provides an important baseline knowledge of the biomagnification dynamics and pathways of Hg in this environment for these multiple interacting species.

Mercury concentrations in marine species from the Aleutian Islands: Spatial and biological determinants.

Several species found in the Bering Sea show significant spatial variation in total mercury concentrations ([THg]) longitudinally along the Aleutian Island chain. We assessed [THg] in other members of the Bering Sea food web to better understand the factors shaping regional differences. [THg] and stable carbon and nitrogen isotope ratios (δ15N and δ13C values) were measured in muscle tissue from 1052 fishes and cephalopods from parts of the Bering Sea and North Pacific Ocean adjacent to the Aleutian Islands. The spatial distribution of the samples enabled regional comparisons for 8 species of fish and one species of cephalopod. Four species showed higher mean length-standardized [THg] in the western Aleutian Islands management area. [THg] in yellow Irish lord were very different relative to those observed in other species and when included in multi-species analyses drove the overall regional trends in mean [THg]. Multi-species analyses excluding measurements for yellow Irish lord showed mean length-standardized [THg] was greater in the western Aleutian Islands than in the central Aleutian Islands management area. Linear regression of [THg] and δ15N values showed a significant and positive relationship across all species, varying between regions and across species. Isotopic space of all species was significantly different between the western Aleutian Islands and central Aleutian Islands, driven largely by δ13C values. Stable isotope values observed follow the same regional trend of lower trophic taxa reported in the literature, with significantly lower δ13C values in the western Aleutian Islands. We conclude that there are regional differences in carbon and nitrogen stable isotope ecology, as well as species-specific feeding ecology that influence [THg] dynamics in part of the marine food web along the Aleutian Island chain. These regional differences are likely contributors to the observed regional variations of [THg] in some high-level predators found in these regions.

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.

Ecotoxicoparasitology of the gastrointestinal tracts of pinnipeds: the effect of parasites on the potential bioavailability of total mercury (THg).

Acanthocephalans, cestodes, and some species of nematodes acquire nutrients from the lumen contents in the gastrointestinal (GI) tract of their definitive host. These parasites are exposed to toxicants, such as mercury (Hg), through passive or active feeding mechanisms; therefore, the focus of this study was to determine if there is an effect of parasites on the dietary availability of total mercury (THg) within piscivorous pinniped hosts. THg concentrations ([THg]) in selected host tissues, parasites, and GI lumen contents from 22 California sea lions (Zalophus californianus), 15 ringed seals (Phoca hispida), and 4 spotted seals (Phoca largha) were determined. Among all pinnipeds, [THg] in acanthocephalans of the large intestine were significantly higher than concentrations in other samples (host lumen contents, other parasites and host intestinal wall), irrespective of location within the host GI tract. δ15N values of parasites depended both on parasite group and location within the GI tract. δ15N values were consistently higher in parasites inhabiting the large intestine, compared to elsewhere in the GI tract, for both sea lions and seals. δ13C values in parasites did not differ significantly from host GI tissues. Based on both [THg] and stable isotope values, parasites are likely affecting the Hg bioavailability within the GI lumen contents and host tissues, and toxicant-parasite interactions appear to depend on both parasitic taxon as well as their location within the host intestine.

Mercury concentrations in three ray species from the Pacific coast of Baja California Sur, Mexico: Variations by tissue type, sex and length.

Total mercury concentrations ([THg]) were determined in muscle and liver of the bat ray (Myliobatis californica), shovelnose guitarfish (Pseudobatos productus) and banded guitarfish (Zapteryx exasperata). Generalized linear models (GLM) were used to determine the effects of size and sex in [THg] and showed that both are determinants of [THg] in these species. The [THg] in both tissues significantly increased with length especially in sexually mature organisms with a steeper slope for mature male than mature female. This may relate to elasmobranchs sexual dimorphism driven variation in growth rates. Median muscle [THg] was significantly greater than liver in each ray species but there were some individuals with higher liver [THg] than muscle. There were individuals with muscle [THg] higher than the advisory thresholds of 0.2 and 0.5mgkg-1ww (2.4 and 11% of the bat ray; 2.1 and 10% of the shovelnose guitarfish; 12.6 and 45% of the banded guitarfish, respectively).

Use of Blood-soaked Cellulose Filter Paper for Measuring Carbon and Nitrogen Stable Isotopes.

We explored the use of filter paper soaked in whole blood for measuring carbon (C) and nitrogen (N) stable isotopes, often used in feeding ecology or diet studies, to better understand drivers of exposure to contaminants. Our results showed no statistically or biologically relevant differences in C and N stable isotope measures between our gold standard (whole blood with anticoagulant) and eluates from processed, blood-soaked filter paper. Our data supported the effectiveness of using filter paper for assessing C and N stable isotopes in blood to address feeding ecology and other uses. The ease of sampling and processing should allow blood-soaked filter paper to be used in sampling of live (e.g., captured, stranded) and lethally taken (e.g., hunter-killed) wild vertebrates.

Nanospray liquid chromatography/tandem mass spectrometry analysis of steroids from gray whale blubber.

RATIONALE: Analysis of steroids from precious blubber biopsies obtained from marine mammals, especially endangered species, can provide valuable information on their endocrine status. Challenges with currently used ELISA methodology include lack of absolute quantitation and incompatibility with multiple steroids analysis due to limited biopsy mass. Development of a sensitive, accurate analytical method for this purpose is critical. METHODS: A nanospray liquid chromatography/tandem mass spectrometry (nanoLC/MS/MS) method was validated for sensitive, specific and quantitative analysis of three steroid hormones, without derivatization, extracted from 50 mg blubber samples. Data was acquired with an LTQ XL ion trap mass spectrometer in positive ion mode, using single reaction monitoring. All three steroids were analyzed in a single run. Cholic acid was used as a surrogate internal standard for quantitation due to its steroidal structure and lack of measurable endogenous levels in blubber. RESULTS: The lowest limits of quantitation for progesterone, testosterone, and hydrocortisone were significantly improved compared to previous studies using conventional LC/MS/MS. The lowest limit of detection was 7 fg/µL using a 1 µL injection volume. Calibration curves for steroid quantification showed good linearity (r2 >0.99) between 14 and 3620 fg/µL, and accuracy was <20% for interday and <10% for intraday. After validation, the method was successfully applied to quantification of steroids in gray whale blubber samples. CONCLUSIONS: The nanoLC/MS/MS method is more sensitive than traditional LC/MS/MS for steroid analysis. It is also compatible with other important biopsy analyses due to its small blubber mass requirement. This will benefit the reproductive and stress assessments for all marine mammals, particularly endangered populations. Copyright © 2017 John Wiley & Sons, Ltd.

Mercury, selenium and fish oils in marine food webs and implications for human health.

Humans who eat fish are exposed to mixtures of healthful nutrients and harmful contaminants that are influenced by environmental and ecological factors. Marine fisheries are composed of a multitude of species with varying life histories, and harvested in oceans, coastal waters and estuaries where environmental and ecological conditions determine fish exposure to both nutrients and contaminants. Many of these nutrients and contaminants are thought to influence similar health outcomes (i.e., neurological, cardiovascular, immunological systems). Therefore, our understanding of the risks and benefits of consuming seafood require balanced assessments of contaminants and nutrients found in fish and shellfish. In this paper, we review some of the reported benefits of fish consumption with a focus on the potential hazards of mercury exposure, and compare the environmental variability of fish oils, selenium and mercury in fish. A major scientific gap identified is that fish tissue concentrations are rarely measured for both contaminants and nutrients across a range of species and geographic regions. Interpreting the implications of seafood for human health will require a better understanding of these multiple exposures, particularly as environmental conditions in the oceans change.