Search

Microplastics (MPs,5 mm in length) have been identified as emerging contaminants in marine environments, with ingestion by a variety of biota being of increasing concern. Few studies exist on MP ingestion in Arctic fish, and there are currently no such data from the Beaufort Sea. We investigated MP abundance in five ecologically valuable species from three sampling sites in the Eastern Beaufort Sea to evaluate possible trophic-level pathways of MPs from prey to beluga whales. Polymer analysis confirmed that 21% of fish gastrointestinal tracts (n = 116) contained microplastic particles. Fish that contained MPs had a mean abundance of 1.42 ± 0.44 particles per individual and an overall mean abundance of 0.37 ± 0.16 particles. No plastic particles5 mm were found, and 78% of the particles observed were fibers. Based on energetic needs, we estimate that individual beluga may ingest between 3800 and 145,000 microplastics annually through trophic transfer, with uncertain health implications.

We assessed the potential role played by two vital Northeastern Pacific Ocean forage fishes, the Pacific sand lance (Ammodytes personatus) and Pacific herring (Clupea pallasii), as conduits for the vertical transfer of microfibres in food webs. We quantified the number of microfibres found in the stomachs of 734 sand lance and 205 herring that had been captured by an abundant seabird, the rhinoceros auklet (Cerorhinca monocerata). Sampling took place on six widely-dispersed breeding colonies in British Columbia, Canada, and Washington State, USA, over one to eight years. The North Pacific Ocean is a global hotspot for pollution, yet few sand lance (1.5%) or herring (2.0%) had ingested microfibres. In addition, there was no systematic relationship between the prevalence of microplastics in the fish stomachs vs. in waters around three of our study colonies (measured in an earlier study). Sampling at a single site (Protection Island, WA) in a single year (2016) yielded most (sand lance) or all (herring) of the microfibres recovered over the 30 colony-years of sampling involved in this study, yet no microfibres had been recovered there, in either species, in the previous year. We thus found no evidence that sand lance and herring currently act as major food-web conduits for microfibres along British Columbia's outer coast, nor that the local at-sea density of plastic necessarily determines how much plastic enters marine food webs via zooplanktivores. Extensive urban development around the Salish Sea probably explains the elevated microfibre loads in fishes collected on Protection Island, but we cannot account for the between-year variation. Nonetheless, the existence of such marked interannual variation indicates the importance of measuring year-to-year variation in microfibre pollution both at sea and in marine biota.

Marine mammals are inherently vulnerable to oil spills. We developed a conceptual framework to evaluate the impacts of potential oil exposure on marine mammals and applied it to 21 species inhabiting coastal British Columbia (BC), Canada. Oil spill vulnerability was determined by examining both the likelihood of species-specific (individual) oil exposure and the consequent likelihood of population-level effects. Oil exposure pathways, ecology, and physiological characteristics were first used to assign species-specific vulnerability rankings. Baleen whales were found to be highly vulnerable due to blowhole breathing, surface filter feeding, and invertebrate prey. Sea otters (Enhydra lutris) were ranked as highly vulnerable due to their time spent at the ocean surface, dense pelage, and benthic feeding techniques. Species-specific vulnerabilities were considered to estimate the likelihood of population-level effects occurring after oil exposure. Killer whale (Orcinus orca) populations were deemed at highest risk due to small population sizes, complex social structure, long lives, slow reproductive turnover, and dietary specialization. Finally, we related the species-specific and population-level vulnerabilities. In BC, vulnerability was deemed highest for Northern and Southern Resident killer whales and sea otters, followed by Bigg's killer whales and Steller sea lions (Eumetopias jubatus). Our findings challenge the typical "indicator species" approach routinely used and underscore the need to examine marine mammals at a species and population level for risk-based oil spill predictions. This conceptual framework can be combined with spill probabilities and volumes to develop more robust risk assessments and may be applied elsewhere to identify vulnerability themes for marine mammals.

As long-lived marine mammals found throughout the temperate coastal waters of the North Pacific and Atlantic Oceans, harbour seals (Phoca vitulina) have become an invaluable sentinel of food-web contamination. Their relatively high trophic position predisposes harbour seals to the accumulation of harmful levels of persistent organic pollutants (POPs). We obtained skin/blubber biopsy samples from live-captured young harbour seals from various sites in the northeastern Pacific (British Columbia, Canada, and Washington State, USA) as well as the northwestern Atlantic (Newfoundland and Quebec, Canada). We developed harbour seal-specific primers to investigate the potential impact of POP exposure on the expression of eight important genes. We found correlations between the blubber mRNA levels of three of our eight target genes and the dominant persistent organic pollutant in seals [polychlorinated biphenyls (PCBs)] including estrogen receptor alpha (Esr1: r 2 = 0.12, p = 0.038), thyroid hormone receptor alpha (Thra: r 2 = 0.16; p = 0.028), and glucocorticoid receptor (Nr3c1: r 2 = 0.12; p = 0.049). Age, sex, weight, and length were not confounding factors on the expression of genes. Although the population-level consequences are unclear, our results suggest that PCBs are associated with alterations of the expression of genes responsible for aspects of metabolism, growth and development, and immune function. Collectively, these results provide additional support for the use of harbour seals as indicators of coastal food-web contamination.

The effects of 2 currently used commercial pesticide formulations on Pacific sockeye salmon (Oncorhynchus nerka), from fertilization to emergence, were evaluated in a gravel-bed flume incubator that simulated a natural streambed. Embryos were exposed to atrazine at 25 µg/L (low atrazine) or atrazine at 250 µg/L (high atrazine) active ingredient (a.i.), and chlorothalonil at 0.5 µg/L (low chlorothalonil) or chlorothalonil at 5 µg/L a.i. (high chlorothalonil) and examined for effects on developmental success and timing, as well as physical and biochemical growth parameters. Survival to hatch was reduced in the high chlorothalonil group (55% compared with 83% in controls), accompanied by a 24% increase in finfold deformity incidence. Reduced alevin condition factor (2.9-5.4%) at emergence and elevated triglyceride levels were seen in chlorothalonil-exposed fish. Atrazine exposure caused premature hatch (average high atrazine time to 50% hatch [H50] = 100 d postfertilization [dpf]), and chlorothalonil exposure caused delayed hatch (high chlorothalonil H50 = 108 dpf; controls H50 = 102 dpf). All treatments caused premature emergence (average time to 50% emergence [E50]: control E50 = 181 dpf, low chlorothalonil E50 = 175 dpf, high chlorothalonil E50 = 174 dpf, high atrazine E50 = 175 dpf, low atrazine E50 = 174 dpf), highlighting the importance of using a gravel-bed incubator to examine this subtle, but critical endpoint. These alterations indicate that atrazine and chlorothalonil could affect survival of early life stages of sockeye salmon in the wild. Environ Toxicol Chem 2017;36:1354-1364. © 2017 SETAC.

Marine plastic pollution is an emerging global conservation challenge, potentially impacting organisms at all trophic levels. However, currently it is unclear to what extent plastic pollution is impacting marine organisms at the population, species or multispecies level. In this study, we explore seasonal exposure (i.e., vulnerability) of Cassin's Auklet (Ptychoramphus aleuticus) to plastic pollution with exposure models during boreal summer and winter seasons. Based on these models, we infer exposure at the population level for this species, in the Canadian Pacific region where approximately 75% of the global population of this species breeds. The models quantify plastic exposure by determining seasonal core foraging areas and plastic concentrations found in those same areas. Core foraging areas were determined using a Generalized Additive Model based on at-sea observation data (collected year round: 1990–2010) and 50% Home Range Kernels based on aerial telemetry data (May and June 1999–2001). Plastic concentrations within these core areas were interpolated based on seawater microplastic concentrations from the summer of 2012. We found that during the boreal summer, Cassin's Auklets were exposed to relatively low concentrations of plastics. During the winter, auklet distribution shifted towards the coast where plastic concentrations are considerably higher. Model derived seasonal variability in exposure was consistent with necropsy results from bird carcasses recovered during the winter of 2014, and from a multiyear study on chick provisioning during the summer. Local oceanography likely plays a role in determining seasonal shifts in both marine bird as well microplastic concentrations, and hence exposure. As well, individual sensitivity (i.e., dose-dependent effect) may vary with annual cycles. Currently, research is focusing on determining how sensitive individual birds are to microplastic concentrations, and our models will help translate sensitivity found at the individual level to potential impacts at population or species level.

Microplastics (MPs, particles

This research was supported by grants to J.-P.D. from the Canadian National Science and Engineering Research Council (NSERC) (PGSD3-443700-2013) and Aarhus University’s Graduate School and Science and Technology (GSST) and Department of Bioscience; and by funding from the Danish DANCEA program (MST-112-00171 and MST-112-00199); the Defra, Scottish and Welsh Governments (for CSIP/SMASS/CEFAS); and the Icelandic Research Fund (i. Rannsóknasjóður; grant no. 120248042). B.M. was supported by funding from NERC (grant no. SMRU 10001). This paper is a contribution from the BONUS BALTHEALTH project, which has received funding from BONUS (Art. 185), funded jointly by the EU, Innovation Fund Denmark, Forschungszentrum Jülich GmbH, German Federal Ministry of Education and Research (grant no. FKZ 03F0767A), Academy of Finland (decision no. 311966), and Swedish Foundation for Strategic Environmental Research. Killer whales (Orcinus orca) are among the most highly polychlorinated biphenyl (PCB)–contaminated mammals in the world, raising concern about the health consequences of current PCB exposures. Using an individual-based model framework and globally available data on PCB concentrations in killer whale tissues, we show that PCB-mediated effects on reproduction and immune function threaten the long-term viability of >50% of the world’s killer whale populations. PCB-mediated effects over the coming 100 years predicted that killer whale populations near industrialized regions, and those feeding at high trophic levels regardless of location, are at high risk of population collapse. Despite a near-global ban of PCBs more than 30 years ago, the world’s killer whales illustrate the troubling persistence of this chemical class. Postprint

Ideally, CD8+ T-cell responses against virally infected or malignant cells are defined at the level of the specific peptide and restricting MHC class I element, a determination not yet made in the dog. To advance the discovery of canine CTL epitopes, we sought to determine whether a putative classical MHC class Ia gene, Dog Leukocyte Antigen (DLA)-88, presents peptides from a viral pathogen, canine distemper virus (CDV). To investigate this possibility, DLA-88*508:01, an allele prevalent in Golden Retrievers, was expressed as a FLAG-tagged construct in canine histiocytic cells to allow affinity purification of peptide-DLA-88 complexes and subsequent elution of bound peptides. Pattern analysis of self peptide sequences, which were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS), permitted binding preferences to be inferred. DLA-88*508:01 binds peptides that are 9-to-12 amino acids in length, with a modest preference for 9- and 11-mers. Hydrophobic residues are favored at positions 2 and 3, as are K, R or F residues at the C-terminus. Testing motif-matched and -unmatched synthetic peptides via peptide-MHC surface stabilization assay using a DLA-88*508:01-transfected, TAP-deficient RMA-S line supported these conclusions. With CDV infection, 22 viral peptides ranging from 9-to-12 residues in length were identified in DLA-88*508:01 eluates by LC-MS/MS. Combined motif analysis and surface stabilization assay data suggested that 11 of these 22 peptides, derived from CDV fusion, hemagglutinin, large polymerase, matrix, nucleocapsid, phosphoprotein, and V proteins, were processed and presented, and thus, potential targets of anti-viral CTL in DLA-88*508:01-bearing dogs. The presentation of diverse self and viral peptides indicates that DLA-88 is a classical MHC class Ia gene.

The Galapagos Islands are one of the last frontiers for conducting ecotoxicologic research because this UNESCO-World Heritage region is not immune to the global and local pollution by chemical contaminants, despite its oceanic remoteness. Endemic marine mammals of the Galapagos are facing several anthropogenic threats and chemical assaults, impacting their populations and habitats. With the aim to assess chemical pollution in the Galapagos Islands, we considered in this appraisal the Galapagos sea lion ( Zalophus wollebaeki ), and Galapagos fur seal ( Arctocephalus galapagoensis ) as the “ canaries in the coal mine ” to monitor the ecosystem health of the Galapagos Archipelago. Special focus is dedicated to the exposure to and biomagnification of persistent organic pollutants, with emphasis on dichlorodiphenyltrichloroethanes. A glimpse of anthropogenic contaminants of emerging concern, requiring urgent research in the Galapagos, is also provided to echo the looming threat of these chemicals in the Galapagos Marine Reserve. Insights on pollutant management and conservation implications in concert with international policy instruments to address pollution are discussed.

Beluga whales (Delphinapterus leucas) are facing profound changes in their habitat, with impacts expected at the individual and population level. Detecting and monitoring exposure and response to environmental stressors is necessary for beluga conservation and management of human activities. Cortisol has proven as a useful tool to assess stress on wildlife. Cortisol was measured in three blubber layers and plasma in subsistence-hunted beluga whales from the summers of 2007 to 2010 using an HPLC/MS/MS. We assessed the effect of biological and biochemical factors. Cortisol ranged from undetectable to 17.8 ng/g in blubber and 2.5 to 61.2 ng/mL in plasma. Concentrations were highest in the inner blubber layer likely reflecting circulating levels. All tissues were significantly higher in 2008 for reasons that remain unclear. Cortisol levels were on par with resting levels in captive belugas. Best fit models for cortisol revealed age to be an important determinant along with length and blubber thickness. Lack o...

Causal evidence linking toxic injury to polychlorinated biphenyl (PCB) exposure is typically confounded by the complexity of real-world contaminant mixtures to which aquatic wildlife are exposed. A local PCB "hotspot" on the Labrador coast provided a rare opportunity to evaluate the effects of PCBs on the health of a marine mammal as this chemical dominated their persistent organic pollutant (POP) burdens. The release of approximately 260 kg of PCBs by a military radar facility over a 30 year period (1970-2000) contaminated some local marine biota, including the ringed seal (Pusa hispida). The abundance profiles of eight health-related gene transcripts were evaluated in liver samples collected from 43 ringed seals in the affected area. The mRNA transcript levels of five gene targets, including aryl hydrocarbon receptor (Ahr), interleukin-1 β (Il1b), estrogen receptor α (Esr1), insulin like growth factor receptor 1 (Igf1), and glucocorticoid receptor α (Nr3c1) correlated with increasing levels of blubber PCBs. PCB threshold values calculated using best-fit hockey-stick regression models for these five genes averaged 1,680 ± 206 ng/g lw, with the lowest, most conservative, being 1,370 ng/g lw for Il1b. Approximately 14% of the seals in the region exceeded this threshold. The dominance of PCBs in the seals studied enabled an assessment of the effects of this chemical on gene transcripts involved in regulating the health of a highly mobile predator, something that is rarely possible in the world of complex mixtures.

Biological and fisheries data were analysed to assess the impact of fisheries mortality on a Critically Endangered subpopulation of 30% of the ETS Sousa subpopulation exhibited injuries caused by fishing gear. Three individuals were photographed with fishing gear attached to their bodies, and 1 dolphin was found dead with fresh injuries caused by fishing gear. To ensure recovery of ETS Sousa, mortality due to human causes should be reduced to

Spectacled caiman (Caiman crocodilus) are fish-eating crocodilians that inhabit freshwater habitat in tropical regions of the Americas. To assess the exposure of caiman to pesticides from banana plantations, the authors collected whole blood samples (30 mL) from 14 adult caiman that were captured in the North Atlantic region of Costa Rica. Blood samples were analyzed for 70 legacy- and current-use pesticides and breakdown products using newly developed ultra-trace, high-resolution mass spectrometry (HRMS). Caiman accumulated pesticides ranked by concentration as dieldrin > permethrin > mirex > 4,4′-DDE > alpha-endosulfan > heptachlor epoxide > oxychlordane > heptachlor > cypermethrin. Caiman within the high-intensity banana crop watershed of Rio Suerte had higher pesticide burdens relative to other more remote locations (F = 12.79; p = 0.00). Pesticide concentration decreased with distance from upstream banana plantations in this river system (F = 20.76; p = 0.00). Caiman body condition was negatively correlated with total pesticide concentrations (F = 6.23; p = 0.02) and with proximity to banana plantations (F = 5.05; p = 0.04). This suggests that either pesticides elicited toxic effects in caiman, resulting in diminished overall health, or that the quantity or quality of their prey was reduced by pesticides downstream of plantation waterways. The authors' results indicate that pesticide use in banana plantations is impacting a high trophic level species inhabiting one of the most important wilderness areas in Costa Rica (Tortuguero National Park). Environ Toxicol Chem 2013;32:2576–2583. © 2013 SETAC

As high trophic level, non-migratory marine mammals, harbor seals (Phoca vitulina) inhabiting the Strait of Georgia, Juan de Fuca Strait and Puget Sound (collectively referred to as the Salish Sea) in northwestern North America provide an integrated measure of coastal food web contamination. We measured congener-specific polychlorinated biphenyls (PCBs), polybrominated diphenylethers (PBDEs), polychlorinated diphenylethers (PCDEs) and polychlorinated naphthalenes (PCNs) in blubber biopsies from free-ranging harbor seal pups inhabiting four sites in the Salish Sea in 2003. While legacy PCBs dominated the composition of these contaminants in seals at all sites (PCBs > PBDEs > PCDEs > PCNs), PBDEs were noteworthy in that they averaged as much as 59% of total PCB concentrations. We further evaluated temporal trends in seals sampled at one of these sites (Puget Sound) for PCBs and PBDEs between 1984 and 2009, and for PCDEs and PCNs between 1984 and 2003. PBDE concentrations doubled every 3.1 years between 1984 and 2003, but appeared to decline thereafter. Over the course of the 20 years between 1984 and 2003, PCB concentrations had declined by 81%, PCDEs declined by 71%, and PCNs by 98%. Overall, results suggest that regulations and source controls have noticeably reduced inputs of these contaminants to the Salish Sea, consequently reducing the associated health risks to marine wildlife. We estimate the total mass of these contaminants in the 53,000 harbor seals of the Salish Sea in 2009 to be 2.6 kg PCBs and 1.0 kg PBDEs, compared to just trace amounts of the PCDEs and PCNs.

Using the feline immunodeficiency virus (FIV) model for AIDS-lentivirus infection, our laboratory has previously demonstrated that T regulatory (Treg) cell-mediated immune T and B cell dysfunction contributes to lentivirus persistence and chronic disease through membrane bound transforming growth factor beta (mTGFb). Studying Treg cells in the context of infection has been problematic as no inducible marker for activated Treg cells had been identified. However, recent reports in human Treg studies have described a novel protein, glycoprotein A repetitions predominant (GARP), as a unique marker of activated human Treg cells that anchors mTGFb. Herein we extend these studies to the feline Treg system, identifying feline GARP and demonstrating that human and feline GARP proteins are homologous in structure, expression pattern, and ability to form a complex with TGFb. We further demonstrate that GARP and TGFb form a complex on the surface of activated Treg cells and that these GARP(+)TGFb(+) Treg cells are highly efficient suppressor cells. Analysis of expression of this Treg activation marker in the FIV-AIDS model reveals an up-regulation of GARP expressing Treg cells during chronic FIV infection. We demonstrate that the GARP(+) Treg cells from FIV-infected cats suppress T helper cells in vivo and that blocking GARP or TGFb eliminates this suppression. These data suggest that GARP is expressed in complex with TGFb on the surface of activated Treg cells and plays an important role in TGFb(+) Treg-mediated T cell immune suppression during lentivirus infection.

While the lethal toxicity of pyrethroid insecticides to fish is well documented, their sublethal physio-behavioral effects remain poorly characterized. Known pyrethroid-associated changes to insect neuromuscular function may translate into similar effects in fish, thereby altering swimming ability and affecting foraging, predator avoidance, and migration. Three experiments were conducted using critical (Ucrit) and burst (Umax) swimming speeds to assess the sublethal effects of the pyrethroids permethrin and deltamethrin in juvenile rainbow trout (Oncorhynchus mykiss). Fish were exposed to deltamethrin (100, 200, or 300 ng/L) or permethrin (1, 2, or 3 μg/L) in water for 4 d, and assessed for swimming performance. Deltamethrin (200 and 300 ng/L) reduced Ucrit, but not Umax, while both swim performance measurements were unaffected by permethrin. Subsequent experiments used only Ucrit to assess deltamethrin exposure. In a time course experiment, deltamethrin (300 ng/L) reduced Ucrit after 1 and 4 d of exposure, but after 7 d of exposure Ucrit was fully recovered. Finally, deltamethrin (1, 2, or 3 μg/L) reduced Ucrit after 1h bath exposures similar to recommended protocols for deltamethrin based sea-lice treatment in aquaculture. The real-world implications of the revealed pyrethroid-associated swimming ability reductions in salmon may be important in areas close to aquaculture facilities.

While the accumulation of persistent contaminants in marine mammals can be attributed directly to their prey, the role of metabolism in shaping patterns is often overlooked. In the present study, the authors investigated the role of metabolic transformation in influencing polychlorinated biphenyl (PCB) and polybrominated diphenyl ether (PBDE) patterns in offshore and nearshore groups of beluga whales (Delphinapterus leucas) and their prey. Congener profiles and principal components analysis (PCA) revealed similar PCB and PBDE patterns in beluga whales feeding either offshore or nearshore, despite divergent contaminant patterns in the putative prey of these two feeding groups. The clustering of PCBs into metabolically derived structure-activity groups (SAGs) and the separation of metabolizable and recalcitrant groups along principal component 1 of the PCA revealed the important role of metabolic transformation in shaping PCB patterns in beluga. Lack of metabolism for congeners with high ortho-chlorine content was revealed by metabolic slopes equal to or greater than 1.0. Metabolic slopes for all other SAGs were less than 1.0 (p

Shellfish and sediment invertebrates have been widely used to assess pollution trends over space and time in coastal environments around the world. However, few studies have compared the bioaccumulation potential of different test species over a range of sediment-contaminant concentrations and profiles. The bioavailability of sediment-related contaminants was evaluated using sediments collected from sites (n = 12) throughout the Salish Sea, British Columbia, Canada. Two benthic marine invertebrates—the Baltic clam Macoma balthica and the polychaete worm Neanthes arenaceodentata—were exposed for 28 days in a controlled environment to these field-collected coastal sediments. The congener-specific uptake of legacy polychlorinated biphenyls (PCBs) and emergent polybrominated diphenyl ethers (PBDEs) was determined using high-resolution gas chromatography/mass spectrometry in sediments and in invertebrates after the experimental exposure. The polychaete Neanthes accumulated lower concentrations of PCBs but higher concentrations of PBDEs. The present study indicates that differences in bioaccumulation between these two invertebrates shape the accumulation of PCB and PBDE congeners, reflect differences in feeding strategies, and reveal the physicochemical properties of the contaminants and sediment properties. Because biota–sediment accumulation factor values are often calculated for environmental monitoring or site-specific impact assessments, our results provide insight into potentially confounding factors and the need for caution when selecting indicator species for coastal marine pollution.

Resistance to Escherichia coli l-asparaginase in canine lymphoma occurs frequently with repeated administration, a phenomenon often attributed, without substantiation, to the induction of neutralizing antibodies. To test the hypothesis that treated dogs develop antibodies against the drug, we created an enzyme-linked immunosorbent assay (ELISA) to measure plasma anti-asparaginase immunoglobulin G responses. Using samples from dogs that had received multiple doses, specific reactivity against l-asparaginase was demonstrated, while naive patients' samples were negative. The optimized ELISA appeared sensitive, with endpoint titers >1 600 000 in positive control dogs. Intra- and inter-assay coefficients of variation were 3.6 and 14.5%. The assay was supported by the observation that ELISA-positive plasma could immunoprecipitate asparaginase activity. When clinical patients were evaluated, 3/10 dogs developed titers after a single injection; with repeated administration, 4/7 dogs were positive. l-asparaginase antibodies showed reduced binding to the PEGylated drug formulation. The ELISA should prove useful in investigating the potential correlation of antibody responses with resistance.

Identifying immunodominant CTL epitopes is essential for studying CD8+ T-cell responses in populations, but remains difficult, as peptides within antigens typically are too numerous for all to be synthesized and screened. Instead, to facilitate discovery, in silico scanning of proteins for sequences that match the motif, or binding preferences, of the restricting MHC class I allele – the largest determinant of immunodominance – can be used to predict likely candidates. The high false positive rate with this analysis ideally requires binding confirmation, which is obtained routinely by an assay using cell lines such as RMA-S that have defective transporter associated with antigen processing (TAP) machinery, and consequently, few surface class I molecules. The stabilization and resultant increased life-span of peptide-MHC complexes on the cell surface by the addition of true binders validates their identity. To determine whether a similar assay could be developed for dogs, we transfected a prevalent class I allele, DLA-88*50801, into RMA-S. In the BARC3 clone, the recombinant heavy chain was associated with murine β2-microglobulin, and importantly, could differentiate motif-matched and -mismatched peptides by surface MHC stabilization. This work demonstrates the potential to use RMA-S cells transfected with canine alleles as a tool for CTL epitope discovery in this species.

For many current use pesticides, limited information exists on their cytotoxicity and immunotoxicity in non-target organisms such as fish. We examined the effects of atrazine, permethrin and piperonyl butoxide (PBO) exposure, in vitro, on rainbow trout (Oncorhynchus mykiss) lymphocyte viability and proliferation. Purified rainbow trout peripheral blood leukocytes (PBLs) were exposed in vitro to the test chemicals (0, 0.01, 0.1, 1 and 10 μM) for 96 h, with and without the mitogen lipopolysaccharide. All three chemicals caused a decrease in both lymphocyte viability and proliferation at 10 μM, while atrazine also suppressed proliferation of PBLs at 1 μM. The in vitro toxicity of these chemicals to this salmonid underscores the need for further investigation using in vivo studies and host resistance models.

While xenoestrogens are routinely detected in the aquatic environment, there is little understanding of the immunotoxicological consequences of exposure to these chemicals in fish, or of the mechanisms through which these effects are mediated. This study was conducted to determine if estrogen receptors (ERs) are present in fish leukocytes and to characterize the effects of 17β-estradiol (E2) and the xenoestrogen nonylphenol (NP) on immune system endpoints in rainbow trout (Oncorhynchus mykiss). Q-PCR was used to confirm that freshly isolated peripheral blood leukocytes (PBLs) express ERs. Following 96-h incubations with E2 or NP (1 nM to 10 μM), PBL ER transcription was again examined using Q-PCR and lipopolysaccharide (LPS)-stimulated proliferation was assessed using flow cytometry. While the transcription of all four forms of rainbow trout ER was unaffected by treatment with E2 or NP, transcription of ERα1 and ERα2 was down-regulated following LPS stimulation. Both E2 and NP, at concentrations of ≥ 100 nM and 10 nM respectively, suppressed leukocyte proliferation. This first report of ERs in rainbow trout PBLs suggests a mechanism through with E2 and other xenoestrogens can modulate immune function. These results highlight the potential for xenoestrogens to impact host resistance to pathogens in wild fish populations.

Increasing emissions of anthropogenic mercury represents a growing concern to the health of high trophic level marine mammals. In its organic form, this metal bioaccumulates, and can be toxic to several physiological endpoints, including the immune system. In this study, we (1) evaluated the effects of inorganic mercury (mercuric chloride, HgCl2) and organic mercury (methylmercuric chloride, MeHgCl) on the in vitro function of lymphocytes isolated from the peripheral blood of beluga whales (Delphinapterus leucas); (2) characterized the potential protective effects of sodium selenite (Na2SeO3) on cell proliferation of HgCl2 or MeHgCl-treated beluga whale lymphocytes; and (3) compared these dose-dependent effects to measurements of blood Hg in samples collected from traditionally harvested beluga whales in the western Canadian Arctic. Lymphocyte proliferative responses were reduced following exposure to 1 μM of HgCl2 and 0.33 μM of MeHgCl. Decreased intracellular thiol levels were observed at 10 μM of HgCl2 and 0.33 μM of MeHgCl. Metallothionein induction was noted at 0.33 μM of MeHgCl. Concurrent exposure of Se provided a degree of protection against the highest concentrations of inorganic Hg (3.33 and 10 μM) or organic Hg (10 μM) for T-lymphocytes. This in vitro protection of Se against Hg toxicity to lymphocytes may contribute to the in vivo protection in beluga whales exposed to high Hg concentrations. Current Hg levels in free-ranging beluga whales from the Arctic fall into the range of exposures which elicited effects on lymphocytes in our study, highlighting the potential for effects on host resistance to disease. The implications of a changing Arctic climate on Hg fate in beluga food webs and the consequences for the health of beluga whales remain pressing research needs.

Killer whales in the NE Pacific Ocean are among the world's most PCB-contaminated marine mammals, raising concerns about implications for their health. Sixteen health-related killer whale mRNA transcripts were analyzed in blubber biopsies collected from 35 free-ranging killer whales in British Columbia using real-time quantitative polymerase chain reaction. We observed PCB-related increases in the expression of five gene targets, including the aryl hydrocarbon receptor (AhR; r(2) = 0.83; p < 0.001), thyroid hormone α receptor (TRα; r(2) = 0.64; p < 0.001), estrogen α receptor (ERα; r(2) = 0.70; p < 0.001), interleukin 10 (IL-10; r(2) = 0.74 and 0.68, males and females, respectively; p < 0.001), and metallothionein 1 (MT1; r(2) = 0.58; p < 0.001). Best-fit models indicated that population (dietary preference), age, and sex were not confounding factors, except for IL-10, where males differed from females. While the population-level consequences are unclear, the PCB-associated alterations in mRNA abundance of such pivotal end points provide compelling evidence of adverse physiological effects of persistent environmental contaminants in these endangered killer whales.

0-DDE Health risk abstract We characterize for the first time the presence of DDT and its metabolites in tropical Galapagos sea lions (Zalophus wolleabeki). P DDT concentrations in Galapagos sea lion pups sampled in 2005 and 2008 ranged from 16 to 3070 lg/kg lipid. Concentrations of P DDT in pups in 2008 averaged 525 lg/kg lipid and were 1.9 times higher than that (281 lg/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 0-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.

Dichlorodiphenyltrichloroethane (DDT) is an effective and relatively cheap anti-malaria pesticide which has saved human lives, but also brought Northern wildlife species to the brink of extinction due to its persistence, bioaccumulation in the food-web, and hormone-mimicking characteristics (Carson 1962; Vallack et al. 1998; UNEP 2001; ATSDR 2002; Blus 2003; WHO 2006; Colborn and Carroll 2007; Van den Berg 2008). Thirty-two years after its ban in the US and its inclusion on Schedule 2 of the UN Stockholm Convention on Persistent Organic Pollutants (POPs) because of its damaging health effects in human and wildlife populations (UNEP 2001; ATSDR 2002; WHO 2006; Vallack et al. 1998; Colborn and Carroll 2007; Blus 2003), the WHO has recommitted to the use of the malaria-fighting pesticide DDT in tropical countries because of an increase in malaria cases in tropical countries (e. g., Peru, Bolivia, Paraguay, Brazil, and African nations), where DDT use was halted or low application rates were used (Van den Berg 2008; WHO 2006; Roberts et al. 1997, 2000). Although evolutionary adaptation of the malaria vector has triggered insecticide resistance to DDT (Denholm et al. 2002), the return to an increased reliance on DDT use is testament to DDT’s effectiveness in combating malaria at a low cost. Large volumes of DDT (i.e., 4,000–5,000 tonnes of active ingredient per year) were used in India and Africa countries to overcome the malaria crisis (Van den Berg 2008). However, 50 years of research on DDT in the temperate and arctic regions of the Northern hemisphere have documented that human health benefits go hand in hand with detrimental effects on wildlife populations (Carson 1962; De Guise et al. 1998; Lahvis et al. 1995; Guillette et al. 1994; Blus 2003;). Since wildlife provides life support, sustenance, and economic opportunities through harvesting and ecotourism in many developing tropical countries, the health of wildlife cannot be viewed in isolation of human health. A balance needs to be struck between health benefits and health risks of DDT use. This requires an understanding of the fate, trends, and dose–response relationship for DDT in tropical wildlife. Such information does currently not exist and monitoring and source control programs that can help to document this relationship are as absent in tropical countries to date as they were in the Western world when DDT was first used in the 1940s and 1960s (Roberts et al. 2000; ATSDR 2002; Blus 2003; Colborn and Carroll 2007; Van den Berg 2008). From a global perspective, the protection of coastal food webs from contamination by chemical pollutants is critical to the long term conservation of the biodiversity and native inhabitants residing in unique places of the Earth, including UNESCO Global Heritage sites such as the Galapagos Islands. Coastal waters that are contaminated with persistent chemicals and pathogens can lead to human illness and adverse health, reduced fisheries quality and quantity, and impacts of the health of marine wildlife. This had obvious social and economic consequences. Conversely, coastal waters that are protected from chemical pollutants provide for an abundance of clean fisheries products and wildlife, and essential foundation for the well-being of the local biodiversity, human residents and the ecotourism sector. At the top of the marine-coastal food chain, marine mammals can provide an “integrated” overview of ecosystem health. As aquatic animals, they are also vulnerable to infection by pathogens of terrestrial origin. By documenting the presence of chemical pollutants in this species, we are able to deliver science-based advice to conservationists, managers, regulators, and stakeholders, on the implementation of best management practices. Equivalent to the role of killer whales as global sentinels of pollution in the Northeastern Pacific, the Galapagos sea lion has recently been used as a sentinel of marine pollution and a key indicator of the ecosystem functioning and oceanic-coastal environmental health in Galapagos Islands. Therefore, this synopsis aims to synthesize our recent findings on DDT contamination in Galapagos sea lions and its use as a sentinel-model species of environmental pollution by POPs.

Numbering no more than 100 individuals and facing many threats, the geographically isolated Eastern Taiwan Strait population of Indo-Pacific humpback dolphins (Sousa chinensis) is in peril. The estuarine and coastal waters of central-western Taiwan have historically provided prime habitat for these dolphins, but environmental conditions today bear little resemblance to what they were in the past. 2. The humpback dolphins must share their habitat with thousands of fishing vessels and numerous factories built upon thousands of hectares of reclaimed land. 3. They are exposed to chemicals and sewage released from adjacent terrestrial activities. Noise and disturbance associated with construction, vessel traffic and military activities are features of everyday life for these animals. 4. Measures to slow the pace of habitat deterioration and reduce the many risks to the dolphins are urgently needed. As one practical step in this direction, this paper describes the habitat needs of these small cetaceans so that decision makers will be better equipped to define 'priority habitat' and implement much needed protection measures under the terms of local legislation. 5. The preferred habitat of these dolphins in Taiwan consists of shallow (o30 m), near-shore marine waters with regular freshwater inputs. 6. For such a small, isolated and threatened population, 'priority habitat' should not be limited to areas of particularly intensive dolphin use or high dolphin density, but rather it should encompass the entire area where the animals have been observed (their current 'habitat'), as well as additional coastal areas with similar bio- physical features ('suitable habitat'). Such a precautionary approach is warranted because the loss of only a few individuals could have serious population-level consequences. 7. While conventional socio-economic analysis might suggest that implementing protection measures over an area stretching � 350 km north-south along Taiwan's west coast and � 3 km out to sea would be too 'costly', the loss of this charismatic species from Taiwan's waters would send a troubling message regarding our collective ability to reconcile human activities with environmental sustainability. Copyright r 2010 John Wiley & Sons, Ltd.

Persistent organic pollutants (POPs) have been associated with adverse health effects in marine mammals. However, the complex mixtures to which free-ranging populations are exposed constrain the elucidation of cause-and-effect relationships between specific POPs and the observed health risks. In this study, we 1) assembled data from studies showing polychlorinated biphenyl (PCB)-associated effects on the health of free-ranging harbor seals in the northeastern Pacific Ocean, 2) carried out additional POP analyses on seal samples to broaden the available data on contaminant residues, and 3) derived estimates of individual POPs and their toxic risks. Taken together, these components were used to generate a new toxicity reference value (TRV) for the protection of marine mammal health. In this case study of seals in British Columbia, Canada, and Washington State, USA, PCBs were the single most abundant POP and were correlated with several adverse health effects. PCB exposures consistently exceeded regulatory toxicity thresholds for fish-eating wildlife. Nursing seal pups were at particular risk, reflecting their greatly increased dietary intake of PCBs and their sensitivity to developmental toxicity. Based on the collective evidence obtained, we propose TRVs (consisting of 5% tissue residue concentration and dose) of 1.3 mg/kg lipid weight tissue residue in blubber and 0.05 mg/kg lipid weight tolerable daily intake in prey. Insofar as the TRVs are lower than previously established TRVs and regulatory guidelines, our study highlights the current underestimation of risks associated with PCBs in high-trophic-level wildlife. Integr Environ Assess Manag 2010;6:631–640. © 2010 SETAC

Olfaction conveys critical environmental information to fishes, enabling activities such as mating, locating food, discriminating kin, avoiding predators and homing. All of these behaviors can be impaired or lost as a result of exposure to toxic contaminants in surface waters. Historically, teleost olfaction studies have focused on behavioral responses to anthropogenic contaminants (e.g., avoidance). More recently, there has been a shift towards understanding the underlying mechanisms and functional significance of contaminant-mediated changes in fish olfaction. This includes a consideration of how contaminants affect the olfactory nervous system and, by extension, the downstream physiological and behavioral processes that together comprise a normal response to naturally occurring stimuli (e.g., reproductive priming or releasing pheromones). Numerous studies spanning several species have shown that ecologically relevant exposures to common pollutants such as metals and pesticides can interfere with fish olfaction and disrupt life history processes that determine individual survival and reproductive success. This represents one of the pathways by which toxic chemicals in aquatic habitats may increasingly contribute to the decline and at-risk status of many commercially and ecologically important fish stocks. Despite our emerging understanding of the threats that pollution poses for chemical communication in aquatic communities, many research challenges remain. These include: (1) the determination of specific mechanisms of toxicity in the fish olfactory sensory epithelium; (2) an understanding of the impacts of complex chemical mixtures; (3) the capacity to assess olfactory toxicity in fish in situ; (4) the impacts of toxins on olfactory-mediated behaviors that are still poorly understood for many fish species; and (5) the connections between sublethal effects on individual fish and the long-term viability of wild populations. This review summarizes and integrates studies on fish olfaction-contaminant interactions, including metrics ranging from the molecular to the behavioral, and highlights directions for future research.

"Southern Resident" killer whales include three "pods" (J, K and L) that reside primarily in Puget Sound/Georgia Basin during the spring, summer and fall. This population was listed as "endangered" in the US and Canada following a 20% decline between 1996 and 2001. The current study, using blubber/epidermis biopsy samples, contributes contemporary information about potential factors (i.e., levels of pollutants or changes in diet) that could adversely affect Southern Residents. Carbon and nitrogen stable isotopes indicated J- and L-pod consumed prey from similar trophic levels in 2004/2006 and also showed no evidence for a large shift in the trophic level of prey consumed by L-pod between 1996 and 2004/2006. Sigma PCBs decreased for Southern Residents biopsied in 2004/2006 compared to 1993-1995. Surprisingly, however, a three-year-old male whale (J39) had the highest concentrations of Sigma PBDEs, Sigma HCHs and HCB. POP ratio differences between J- and L-pod suggested that they occupy different ranges in winter.

We previously reported in vivo induction of cytochrome P450 1A (CYP1A) by β-naphthoflavone in skin and liver biopsies of captive harbour seals ( Phoca vitulina richardsi ). The present study evaluated CYP1A expression (immunoblot analysis and ethoxyresorufin O -deethylase activity-EROD) in harbour seals using two study designs: i) skin and liver biopsies from 20 harbour seal pups captured from coastal British Columbia (BC, Canada) and temporarily housed in captivity; and ii) skin biopsies from 42 free-ranging harbour seals captured and sampled on-site in multiple locations in BC and Washington State (USA). Toxic Equivalency Quotients (TEQs) were calculated for polychlorinated biphenyl, polychlorinated dibenzo- p -dioxin, and polychlorinated dibenzofuran residues measured in blubber from a subset of study animals ( n = 30). CYP1A data from the seal pups held temporarily in captivity show that CYP1A protein levels were greater in liver than skin and that CYP1A protein and EROD activity were correlated in skin and liver. However, analysis of free-ranging seals from different sites revealed that blubber organochlorine TEQ values did not correlate with skin CYP1A levels. CYP1A protein levels and EROD activities in skin of seal pups from the BC locations and from Puget Sound were relatively low, possibly reflecting contaminant levels that were not high enough to elicit a response, a small sample size, or methodological limitations. Our results show that CYP1A measurements in skin show promise as a biomarker of contaminant exposure, but that refinements to techniques and a larger sample size are needed.

We hypothesized that depleted fat reserves in grizzly bears (Ursus arctos horribilis) following annual hibernation would reveal increases in persistent organic pollutant (POP) concentrations compared to those present in the fall. We obtained fat and hair from British Columbia grizzly bears in early spring 2004 to compare with those collected in fall 2003, with the two tissue types providing contaminant and dietary information, respectively. By correcting for the individual feeding habits of grizzlies using a stable isotope-based approach, we found that polychlorinated biphenyls (sigmaPCBs) increased by 2.21x, polybrominated diphenylethers (sigmaPBDEs) increased by 1.58x, and chlordanes (sigmaCHL) by 1.49x in fat following hibernation. Interestingly, individual POPs elicited a wide range of hibernation-associated concentration effects (e.g., CB-153, 2.25x vs CB-169, 0.00x), resulting in POP pattern convergence in a PCA model of two distinct fall feeding groups (salmon-eating vs non-salmon-eating) into a single spring (post-hibernation) group. Our results suggest that diet dictates contaminant patterns during a feeding phase, while metabolism drives patterns during a fasting phase. This work suggests a duality of POP-associated health risks to hibernating grizzly bears: (1) increased concentrations of some POPs during hibernation; and (2) a potentially prolonged accumulation of water-soluble, highly reactive POP metabolites, since grizzly bears do not excrete during hibernation.

Odor-evoked neurophysiological responses can form the basis for behavioral responses. Here we first characterized olfactory-mediated behavioral and neurophysiological responses of juvenile rainbow trout to the amino acid l-histidine, then looked at whether there were similar responses to the carbamate antisapstain IPBC and the herbicides atrazine and Roundup, and lastly explored how exposures to these pesticides modified the l-histidine responses. Trout were behaviorally attracted to 10(-7)M l-histidine (as assayed in a counter-current olfactometer), but this preference behavior switched to indifference with higher histidine concentrations. Neurophysiologically, the summed electrical responses of peripheral olfactory neurons, as measured using electro-olfactogram (EOG), was 0.843+/-0.252 mV to 10(-7)M l-histidine. Of the pesticides, only Roundup evoked EOGs, indicating the amino acid-based pesticide may have acted as an odorant, and generated a behavioral response: it was avoided at active ingredient [AI; glyphosate isopropyl amine] concentrations > or =10 mg/l. With 30 min pesticide exposures, 10(-7)M l-histidine preference behavior was eliminated following exposure to 1 microg/l IPBC and atrazine, and 100 microg/l AI Roundup. Similarly, 10(-7)M l-histidine-evoked EOGs were significantly reduced by exposure to 1 microg/l IPBC, 10 microg/l atrazine, and 100 microg/l AI Roundup. When combined together, the results demonstrate that typical preference behavior can be abolished when neurophysiological responses are reduced by >60% of control. This asymmetry in response thresholds suggests that behavioral responses may be more sensitive toxicological endpoints than neurophysiological responses.

Microplastics are increasingly recognized as being widespread in the world's oceans, but relatively little is known about ingestion by marine biota. In light of the potential for microplastic fibers and fragments to be taken up by small marine organisms, we examined plastic ingestion by two foundation species near the base of North Pacific marine food webs, the calanoid copepod Neocalanus cristatus and the euphausiid Euphausia pacifia. We developed an acid digestion method to assess plastic ingestion by individual zooplankton and detected microplastics in both species. Encounter rates resulting from ingestion were 1 particle/every 34 copepods and 1/every 17 euphausiids (euphausiids > copepods; p = 0.01). Consistent with differences in the size selection of food between these two zooplankton species, the ingested particle size was greater in euphausiids (816 ± 108 μm) than in copepods (556 ± 149 μm) (p = 0.014). The contribution of ingested microplastic fibres to total plastic decreased with distance from shore in euphausiids (r (2) = 70, p = 0.003), corresponding to patterns in our previous observations of microplastics in seawater samples from the same locations. This first evidence of microplastic ingestion by marine zooplankton indicate that species at lower trophic levels of the marine food web are mistaking plastic for food, which raises fundamental questions about potential risks to higher trophic level species. One concern is risk to salmon: We estimate that consumption of microplastic-containing zooplankton will lead to the ingestion of 2-7 microplastic particles/day by individual juvenile salmon in coastal British Columbia, and ≤91 microplastic particles/day in returning adults.

The transplacental transfer of persistent organic pollutants in marine mammals takes place at a formative developmental period, thereby exposing the fetus to endocrine-disrupting compounds. We evaluated the transplacental transfer of polychlorinated biphenyls (PCBs), polybrominated diphenylethers (PBDEs), and organochlorine pesticides (OCPs) in five pregnant ringed seals in Northern Labrador, Canada. PCBs, PBDEs, and OCPs were transferred from the mother to the fetus with average concentrations in the fetuses ranging from 0.3 ng/g lipid weight (lw) of mirex to 94 ng/g lw of PCBs. The average percent transferred to the blubber in the fetus was very low with

Resident killer whale populations in the NE Pacific Ocean are at risk due to the accumulation of pollutants, including polybrominated diphenyl ethers (PBDEs). To assess the impact of PBDEs in water and sediments in killer whale critical habitat, we developed a food web bioaccumulation model. The model was designed to estimate PBDE concentrations in killer whales based on PBDE concentrations in sediments and the water column throughout a lifetime of exposure. Calculated and observed PBDE concentrations exceeded the only toxicity reference value available for PBDEs in marine mammals (1500 μg/kg lipid) in southern resident killer whales but not in northern resident killer whales. Temporal trends (1993–2006) for PBDEs observed in southern resident killer whales showed a doubling time of ≈5 years. If current sediment quality guidelines available in Canada for polychlorinated biphenyls are applied to PBDEs, it can be expected that PBDE concentrations in killer whales will exceed available toxicity reference values by a large margin. Model calculations suggest that a PBDE concentration in sediments of approximately 1.0 μg/kg dw produces PBDE concentrations in resident killer whales that are below the current toxicity reference value for 95 % of the population, with this value serving as a precautionary benchmark for a management-based approach to reducing PBDE health risks to killer whales. The food web bioaccumulation model may be a useful risk management tool in support of regulatory protection for killer whales.

Mercury (Hg) bioaccumulates in the aquatic food chain in the form of methylmercury, a compound well known for its neurotoxicity. We analyzed total mercury (THg) in hair collected from 209 harbour seals captured at 10 sites in British Columbia (Canada) and Washington State (USA) between 2003 and 2010. In addition, laser ablation inductively-coupled plasma mass spectrometry (LA-ICP-MS) allowed for a highly refined analysis of THg accumulation over time by examining nine whiskers taken from 4- to 6-week-old pups. We estimate that THg concentrations in pups increased sharply at a point corresponding to mid- to late gestation of their time in utero (4.7 ± 0.8 and 6.6 ± 1.3 µg/g dry weight (dw), respectively), and then again at the onset of nursing (8.1 ± 1.3 µg/g dw). These abrupt changes highlight the importance of both pre- and post-natal THg transfer from the mother to the growing fetus and the newborn pup. While THg levels varied among sites, hair analyses from seals collected at the same site demonstrated the influence of age in THg accumulation with pups (5.3 ± 0.3 µg/g) and juveniles (4.5 ± 0.5 µg/g) having lower levels than those in adults (8.3 ± 0.8 µg/g). Our results revealed that 33 % of the pups sampled (n = 167) had THg levels that surpassed a mammalian hair threshold for neurochemical alterations. This study suggests that Hg could represent a health concern to marine wildlife, especially as atmospheric emissions of this toxic element from human activities in the Pacific Rim and worldwide continue.

Persistent organic pollutants are environmental contaminants that, because of their lipophilic properties and long half-lives, bioaccumulate within aquatic food webs and often reach high concentrations in marine mammals, such as harbor seals (Phoca vitulina). Exposure to these contaminants has been associated with developmental abnormalities, immunotoxicity, and reproductive impairment in marine mammals and other high-trophic-level wildlife, mediated via a disruption of endocrine processes. The highly conserved thyroid hormones (THs) represent one vulnerable endocrine end point that is critical for metabolism, growth, and development in vertebrates. We characterized the relationship between contaminants and specific TH receptor (TR) gene expression in skin/blubber biopsy samples, as well as serum THs, from free-ranging harbor seal pups (n = 39) in British Columbia, Canada, and Washington State, USA. We observed a contaminant-related increase in blubber TR-alpha gene expression [total polychlorinated biphenyls (capital sigmaPCBs); r = 0.679; p0.001] and a concomitant decrease in circulating total thyroxine concentrations (capital sigmaPCBs; r = -0.711; p0.001) . Consistent with results observed in carefully controlled laboratory and captive feeding studies, our findings suggest that the TH system in harbor seals is highly sensitive to disruption by environmental contaminants. Such a disruption not only may lead to adverse effects on growth and development but also could have important ramifications for lipid metabolism and energetics in marine mammals.

Biomarkers of organochlorine exposure, such as the induction of cytochrome P450 1A (CYP1A), can be used to assess the impact of environmental contaminants on the health of free-ranging marine mammal populations. The objective of the present study was to measure CYP1A in skin and liver biopsies obtained from live harbour seals (Phoca vitulina). Twelve harbour seal pups, aged three to five weeks, were captured from the Fraser River estuary, British Columbia, Canada, and temporarily held in captivity. Skin ( approximately 60 mg) and liver ( approximately 40 mg) biopsies, obtained while seals were under general anaesthesia, yielded sufficient tissue for the measurement of CYP1A by immunoblot analysis and ethoxyresorufin O-deethylase activity. A short-term exposure experiment, in which harbour seals (n=3) were treated orally with beta-naphthoflavone (BNF), resulted in increased hepatic and cutaneous CYP1A protein levels, consistent with observations in other mammals. This study is the first to measure CYP1A in skin and liver biopsies from live harbour seals and to report in vivo BNF-associated CYP1A induction in a marine mammal. The results demonstrate that microsamples collected using minimally-invasive techniques can provide toxicologically-relevant information form marine mammals.

Vitamin A is a nutrient essential to all mammals for growth and development, as well as for the maintenance of reproductive, endocrine, and immune systems. Environmental contaminant-related disruption of vitamin A has been observed in many wildlife species and can therefore be used as a biomarker of toxic effects. However, the natural processes regulating vitamin A uptake, storage, and distribution among compartments are poorly understood in marine mammals. In this study, 20 young healthy harbour seals (Phoca vitulina) were captured to establish a compartment-based model providing a foundation for a mechanistic understanding of vitamin A physiology and disruption. Vitamin A (retinol, retinyl palmitate, and (or) retinoic acid) was quantified in blood plasma and in biopsy samples of liver, blubber, and skin. Although the highest concentrations of vitamin A were found in liver, blubber represents a more important storage depot, with an estimated 66% of the total retinoid content of the compartments measured. We suggest that vitamin A physiology in the precocious harbour seal has evolved to deal with high vitamin A availability during a short nursing period and to sustain growth during the postweaning fast. Positive correlations in vitamin A concentrations among liver, blubber, and skin support the use of less invasive biopsy sampling of just blubber or skin, which can provide physiologically relevant information in biomarker studies of free-ranging marine mammals.

Harbor seals (Phoca vitulina) are small pinnipeds that are widely distributed throughout the temperate coastal regions of the Atlantic and Pacific oceans. We determined birth mass, neonatal growth rates, weaning age, and weaning mass of NE Pacific harbor seals (P. v. richardsi) during a capture-recapture study that spanned the nursing period (Sidney Island, British Columbia, Canada). Of 46 harbor seal pups initially captured, 28 were classified as newborns (i. e., < 24 h old). Mean body mass of newborns was 11.2 ± SE 0.31 kg. Pups were individually tagged and recaptured throughout the nursing period. Average daily mass gain during the nursing period was 394 ± 26 g. Mean birth mass of males did not differ significantly from females, although pups found with fetal pelage (lanugo) (21.4% of all newborns) were smaller at birth (9.8 ± 0.44 kg) than non-lanugo pups (11.6 ± 0.33 kg). Mean weaning mass was estimated at 23.6 ± 1.2 kg at a mean weaning age of 32 d ± 1.5 d. While birth and weaning masses differed little from the published data for offshore Sable Island harbor seals (P. v. concolor), British Columbia harbor seals are characterized by half the daily mass gain and a longer nursing period.

A series of high profile outbreaks of newly described diseases in humans, domestic animals and wildlife has attracted widespread interest in the topic of Emerging Infectious Diseases (EIDs). Marine mammals are no exception: since 1987 several mass mortalities have been observed following infection with viruses previously undescribed in the populations or species in question. As with terrestrial examples, some of these outbreaks have followed either migrations associated with large-scale ecological changes or the introduction of virus from domestic animals. However, marine mammals warrant special concern in the context of emerging infectious diseases: they typically occupy high trophic levels and can therefore be highly contaminated with immunotoxic chemicals. Persistent Organic Pollutants (POPs), including polychlorinated -biphenyls (PCBs), dibenzo-p-dioxins (PCDDs), -dibenzofurans (PCDFs) and related compounds, are demonstrated immunotoxicants in laboratory animals, as well as marine mammals. Immunotoxic...

High trophic level arctic beluga whales (Delphinapterus leucas) are exposed to persistent organic pollutants (POP) originating primarily from southern latitudes. We collected samples from 43 male beluga harvested by Inuvialuit hunters (2008-2010) in the Beaufort Sea to evaluate the effects of POPs on the levels of 13 health-related gene transcripts using quantitative real-time polymerase chain reaction. Consistent with their role in detoxification, the aryl hydrocarbon receptor (Ahr) (r(2) = 0.18, p = 0.045 for 2008 and 2009) and cytochrome P450 1A1 (Cyp1a1) (r(2) = 0.20, p0.001 for 2008 and 2009; r(2) = 0.43, p = 0.049 for 2010) transcripts were positively correlated with polychlorinated biphenyls (PCBs), the dominant POP in beluga. Principal Components Analysis distinguished between these two toxicology genes and 11 other genes primarily involved in growth, metabolism, and development. Factor 1 explained 56% of gene profiles, with these latter 11 gene transcripts displaying greater abundance in years coinciding with periods of low sea ice extent (2008 and 2010). δ(13)C results suggested a shift in feeding ecology and/or change in condition of these ice edge-associated beluga whales during these two years. While this provides insight into the legacy of PCBs in a remote environment, the possible impacts of a changing ice climate on the health of beluga underscores the need for long-term studies.

Although vitamin A (retinol) levels are highly regulated within individual organisms, natural (e.g., age, sex, disease) and anthropogenic (e.g., environmental contaminants) factors can affect the dynamics of this essential nutrient. In this study, we examined developmental changes in the circulatory vitamin A system of free-ranging harbour seal (Phoca vitulina) pups by collecting serial blood samples from healthy known-age animals throughout their nursing period. While harbour seal pups were born with relatively low levels of circulatory retinol (144.4 ± 13.9 µg/L), nursing animals more than doubled these levels within 2 days (385.0 ± 46.9 µg/L), and levels continued to rise more gradually until weaning (431.0 ± 35.8 µg/L). Animals that were not nursing, such as orphaned (184.4 ± 34.2 µg/L), fasted (347.0 ± 14.4 µg/L), and weaned (204.5 ± 38.5 µg/L) pups, had significantly lower circulatory retinol levels. Despite the developmental changes observed in total retinol, the concentration of retinol bound by its transport proteins, retinol binding protein and transthyretin, remained relatively constant throughout the nursing period. This suggests that, like most mammals, the delivery of retinol to target tissues is highly regulated in harbour seal pups. Furthermore, the high concentrations of circulatory retinol observed in harbour seal pups may serve to saturate transport proteins, ensuring a steady delivery of vitamin A to target tissues during a period of potentially variable supply. Understanding how natural factors affect circulatory retinol and its transport proteins is an important facet of assessing the impact of environmental contaminants on vitamin A dynamics in marine mammals.

In recent decades, marine mammal populations living in highly polluted areas have experienced incidences of low reproductive success, developmental abnormalities and disease outbreaks. In many of these cases, environmental contaminants were suspected as causal or contributing factors. However, demonstrating a mechanistic link between contaminant exposure and effect in marine mammal populations has proven challenging. Consequently, the development and application of relatively noninvasive biomarkers represents a potentially valuable means of monitoring wildlife populations exposed to elevated levels of contaminants. One touted biomarker is vitamin A (retinol), a “dietary hormone” whose metabolites are required for reproduction, growth, development, immune function, vision and epithelial maintenance. Laboratory studies have shown that many contaminants, including polychlorinated biphenyls (PCBs), polychlorinated dibenzo- para-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), can disrupt vitamin A physiology and alter the distribution of its essential metabolites. Field studies suggest that complex environmental mixtures of these chemicals can also interfere with vitamin A dynamics in free-ranging marine mammals and other fish-eating wildlife. However, circulatory retinol, which is the least invasive measurement of vitamin A status, appears to have variable responses to contaminant exposure. In addition, “normal” circulatory retinol levels have not yet been described for most wildlife species, and not enough is known about the natural physiological events that can alter these concentrations. Confounding factors must therefore be characterized before retinoids can be used as an effective indicator of adverse health effects in marine mammals exposed to elevated levels of environmental contaminants.

Blubber biopsy samples were obtained for contaminant analysis from two discrete populations of killer whales (Orcinus orca) which frequent the coastal waters of British Columbia, Canada. Detailed life history information for the fish-eating ‘resident’ population, comprising two distinct communities, and the marine mammal-eating ‘transient’ killer whale population, provided an invaluable reference for the interpretation of contaminant concentrations. Total PCB concentrations (sum of 136 congeners detected) were surprisingly high in all three communities, but transient killer whales were particularly contaminated. PCB concentrations increased with age in males, but were greatly reduced in reproductively active females. The absence of age, sex and inter-community diAerences in concentrations of polychlorinated- dibenzo-p-dioxins (PCDDs) and- dibenzofurans (PCDFs) may have partly reflected low dietary levels, but more importantly, metabolic removal of dioxin-like compounds in killer whales. While information on toxic thresholds does not exist for PCBs in cetaceans, total 2,3,7,8-TCDD Toxic Equivalents (TEQ) in most killer whales sampled easily surpassed adverse eAects levels established for harbour seals, suggesting that the majority of free-ranging killer whales in this region are at risk for toxic eAects. The southern resident and transient killer whales of British Columbia can now be considered among the most contaminated cetaceans in the world. ” 2000 Elsevier Science Ltd. All rights reserved.

As high trophic level organisms in the marine environment, fish-eating seals, dolphins and whales are often exposed to very high levels of fat-soluble environmental contaminants. Assessing the sources, levels and patterns of contaminants found in the tissues of marine mammals, and the biological effects of these contaminants on individuals, is essential to determining any population- or species-level impacts. While the number of contaminants to which marine mammals are exposed is staggering, designing strategies to assess the effects of complex mixtures represent a challenging yet vital part of an understanding of the “real world”. At present, an accumulated “weight of evidence” suggests that ambient levels of lipophilic contaminants have adversely affected aspects of reproduction, immune function and endocrine function in marine mammals inhabiting a number of industrial coastal regions. This body of evidence is drawn from a combination of (1) epidemiological or descriptive studies of effects observed in ...

While the immunotoxicity of 2,3,7,8-tetrachlo- rodibenzo-p-dioxin (TCDD) has been well established, the eAects of complex environmental mixtures of poly- halogenated aromatic hydrocarbons (PHAHs) are poorly understood. Many PHAHs, including the poly- chlorinated-biphenyls (PCBs), -dibenzofurans (PCDFs), and dibenzo-p-dioxins (PCDDs), possess 'dioxin-like' activities, and accumulate in the aquatic food chain. Organisms occupying high trophic levels may therefore be exposed to concentrations which may present an immunotoxic risk. In this study, pregnant PVG rats were administered a daily oral dose of 1 ml of the following during pregnancy and lactation: (1) oil extracted from herring caught in the relatively uncontaminated Atlantic Ocean; (2) oil extracted from herring caught in the contaminated Baltic Sea; or (3) the Atlantic herring oil extract spiked with 2,3,7,8-TCDD. The daily intakes of aryl hydrocarbon (Ah)-receptor dependent toxic equiv- alents (TEQ) for mothers were 0.3 in the Atlantic group, 2.1 in the Baltic group, and 134 ng/kg body wt. in the 2,3,7,8-TCDD positive control group. Immune function and host resistance to rat cytomegalovirus (RCMV) were assessed in oAspring aged 11, 25, 46 or 59 days. Rat pups in the positive control TCDD-spiked group exhibited immunosuppression characterized by reduced thymus weight and cellularity, reduced thymocyte and splenocyte proliferative responses to T-dependent mi- togens in vitro, reduced virus-associated natural killer (NK) cell and specific antibody responses. While less pronounced, a similar pattern of eAects was observed in the rat pups exposed only to the Baltic Sea herring oil. These immunotoxic eAects were transient in both ex- posure groups, with a time-related recovery in immune function possibly due to the half-life of TCDD in rats and the waning exposure levels in the rapidly growing pups. We previously demonstrated that the same Baltic Sea herring led to impaired natural killer cell and T- lymphocyte function in harbour seals during the course of a long-term captive feeding study. The collective re- sults of these studies in rats and seals indicate the immunotoxic potential of environmental mixtures at current levels in the aquatic environment, and suggest that the developing immune system of young mammals may be at particular risk.