PCBs are associated with altered gene transcript profiles in arctic Beluga Whales (Delphinapterus leucas).

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, p < 0.001 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.

PCB related effects thresholds as derived through gene transcript profiles in locally contaminated ringed seals (Pusa hispida).

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 ß (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.

Persistent organic pollutants in chinook salmon (Oncorhynchus tshawytscha): implications for resident killer whales of British Columbia and adjacent waters.

We measured persistent organic pollutant (POP) concentrations in chinook salmon (Oncorhynchus tshawytscha) in order to characterize dietary exposure in the highly contaminated, salmon-eating northeastern Pacific resident killer whales. We estimate that 97 to 99% of polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), dichlorodiphenyltrichloroethane (DDT), and hexachlorocyclohexane (HCH) in returning adult chinook were acquired during their time at sea. Highest POP concentrations (including PCBs, PCDDs, PCDFs, and DDT) and lowest lipids were observed in the more southerly chinook sampled. While feeding by salmon as they enter some more POP-contaminated near-shore environments inevitably contribute to their contamination, relationships observed between POP patterns and both lipid content and delta13C also suggest a migration-related metabolism and loss of the less-chlorinated PCB congeners. This has implications for killer whales, with the more PCB-contaminated salmon stocks in the south partly explaining the 4.0 to 6.6 times higher estimated daily intake for sigmaPCBs in southern resident killer whales compared to northern residents. We hypothesize that the lower lipid content of southerly chinook stocks may cause southern resident killer whales to increase their salmon consumption by as much as 50%, which would further increase their exposure to POPs.

Assessment of cytochrome P450 1A in harbour seals (Phoca vitulina) using a minimally-invasive biopsy approach.

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.

Biomagnification of polychlorinated biphenyls in a harbor seal (Phoca vitulina) food web from the Strait of Georgia, British Columbia, Canada.

Polychlorinated biphenyl (PCB) biomagnification was characterized in a harbor seal food web in the Strait of Georgia, British Columbia, Canada. Trophic magnification factors (TMFs) for PCBs averaged 3.6, with a range of 0.7 to 9.4. The TMFs for individual congeners correlated with log K(OW) (r(2) = 0.56, p < 0.001), reflecting the role that physicochemical properties play in driving the biomagnification of PCBs in marine food webs. However, TMFs differed among PCB structure activity groups, clearly indicating an additional role for metabolic transformation of certain PCBs. The known feeding preferences of harbor seals enabled the calculation of trophic level-adjusted biomagnification factors (BMF(TL)) for PCBs in this species, which averaged 13.4 and ranged from 0.2 to 150.6. Metabolic transformation in seals explained some of the variation in congener-specific biomagnification, with lower BMF(TL) values for PCB congeners with meta- and parachlorine unsubstituted positions. Principal components analysis revealed the distinct roles played by trophic level, log K(OW), and metabolic transformation in explaining the notable differences in PCB patterns among harbor seals, their pups, and their prey. In the present study, the authors estimate there to be approximately 76 kg of PCBs in the biota of the Strait of Georgia, of which 1.6 kg is retained by harbor seals.

Cytochrome P450 1A expression and organochlorine contaminants in harbour seals (Phoca vitulina): evaluating a biopsy approach.

We previously reported in vivo induction of cytochrome P450 1A (CYP1A) by beta-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.