Polychlorinated Biphenyl-Related Alterations of the Expression of Essential Genes in Harbour Seals (Phoca vitulina) from Coastal Sites in Canada and the United States.

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.

A Risk-Based Characterization of Sediment Contamination by Legacy and Emergent Contaminants of Concern in Coastal British Columbia, Canada.

Sediments have long been used to help describe pollution sources, contaminated sites, trends over time, and habitat quality for marine life. We collected surficial sediments from 12 sites at an average seawater depth of 25 m in three near-urban areas of the Salish Sea (British Columbia, Canada) to investigate habitat quality for marine life, including heavily contaminated killer whales. Samples were analyzed using high-resolution instrumentation for a wide variety of congeners of polychlorinated biphenyls (PCBs), polybrominated diphenylethers (PBDEs), hexabromocyclododecane (HBCDD), polybrominated biphenyls, polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans, organochlorine pesticides, and polychlorinated naphthalenes (PCNs). The top six contaminant classes detected in sediments were ∑PCB > ∑PBDE > ∑PCDD/F > DDT > ∑HBCDD > ∑PCN. Near-urban harbor sediments had up to three orders of magnitude higher concentrations of contaminants than more remote sites. With limited tools available to characterize biological risks associated with complex mixtures in the real world, we applied several available approaches to prioritize the pollutant found in our study: (1) sediment quality guidelines from the Canadian Council of Ministers of the Environment where available; (2) US NOAA effects range low and other international guidelines; (3) total TEQ for dioxin-like PCBs for the protection of mammals; and (4) the calculation of risk quotients. Our findings provide an indication of the state of contamination of coastal environments in British Columbia and guidance for chemical regulations and priority setting, as well as management actions including best-practices, dredging, disposal at sea, and source control. In this regard, the legacy PCB and the emergent PBDEs should command continued priority monitoring.

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.

Heightened biological uptake of polybrominated diphenyl ethers relative to polychlorinated biphenyls near-source revealed by sediment and plankton profiles along a coastal transect in British Columbia.

Polychlorinated biphenyl (PCB) and polybrominated diphenyl ether (PBDE) concentrations and profiles in paired sediment-plankton samples were determined along a 500 km transect in coastal British Columbia, Canada. PCB and PBDE levels in sediment were both greater in the industrialized Strait of Georgia than in remote northern sites and exhibited parallel spatial trends. In plankton, recent-use PBDE levels were higher near-source, while levels of legacy PCBs were uniform across sites. Principal component analysis of 95 PCB congeners illustrated the influence of proximity to source (i.e., latitude) on congener patterns for both matrices (sediment, r(2) = 0.52, p = 0.012; plankton, r(2) = 0.59, p = 0.016). The PCB pattern in plankton grew lighter with latitude, but the opposite pattern in sediments suggested that temperature-related fractionation, sediment processes, and basin-wide oceanography had divergent effects on each matrix. Biota-sediment accumulation factors (BSAFs) were greater for PBDEs than PCBs, but spatial profiles were similar; PCBs and PBDEs were near equilibrium in remote atmospherically driven sites (BSAF = 1.7 and 1.3) but accumulated preferentially in sediments at source-driven sites (BSAF = 0.2 and 0.4). The influences of particle-binding and hydrophobicity on the aquatic fate of PCBs and PBDEs was evident by the strong influence of log KOW on congener-specific BSAFs (PCBs, r(2) = 0.18 p < 0.001; PBDEs, r(2) = 0.61 p < 0.001). While biotic uptake of PCBs has become spatially uniform in coastal BC because of dilution over time, biomagnification of PBDEs remains higher in industrialized waters.

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.

Widespread distribution of microplastics in subsurface seawater in the NE Pacific Ocean.

We document the abundance, composition and distribution of microplastics in sub-surface seawaters of the northeastern Pacific Ocean and coastal British Columbia. Samples were acid-digested and plastics were characterized using light microscopy by type (fibres or fragments) and size (<100, 100-500, 500-100 and >1000 µm). Microplastics concentrations ranged from 8 to 9200 particles/m(3); lowest concentrations were in offshore Pacific waters, and increased 6, 12 and 27-fold in west coast Vancouver Island, Strait of Georgia, and Queen Charlotte Sound, respectively. Fibres accounted for ∼ 75% of particles on average, although nearshore samples had more fibre content than offshore (p<0.05). While elevated microplastic concentrations near urban areas are consistent with land-based sources, the high levels in Queen Charlotte Sound appeared to be the result of oceanographic conditions that trap and concentrate debris. This assessment of microplastics in the NE Pacific is of interest in light of the on-coming debris from the 2011 Tohoku Tsunami.

Vitamin A and E profiles as biomarkers of PCB exposure in beluga whales (Delphinapterus leucas) from the western Canadian Arctic.

We evaluated the utility of vitamin A and E profiles as biomarkers of contaminant exposure in beluga whales (Delphinapterus leucas; n=66) harvested by the Inuvialuit in the Beaufort Sea. Blubber was an important repository for these vitamins, accounting for 76.8±2.6% of the total body store of vitamin A, and 98.5±0.4% of total vitamin E. While the free alcohol form of vitamin A (retinol) appeared highly regulated, the vitamin A esters were influenced by several biological factors including age, body condition and length. Vitamin E concentrations in liver and blubber were related to age, condition, length and feeding ecology, as described δ(15)N and δ(13)C. Despite the influence of these factors, collective results from univariate statistics, best fit multiple regressions, and principal component analysis (PCA) identified polychlorinated biphenyls (PCBs) as important determinants of vitamin concentrations and profiles in beluga tissues. Blubber PCB concentrations best explained variation of the first principal component in a PCA of hepatic vitamins (r(2)=0.13, p=0.014), and regression models found that vitamin A concentrations were negatively correlated with PCB levels in liver (esters: r(2)=0.19, p=0.001), but positively in plasma (retinol: r(2)=0.20, p=0.06) and blubber (retinol: r(2)=0.22, p=0.001, esters: r(2)=0.43, p<0.001). Our analyses provide a basis to propose an integrated toxicity reference value for disruption of vitamin A and E profiles in beluga of 1.6 mg/kg lw PCBs. This disruption of vitamin profiles by moderate levels of PCBs in an arctic cetacean highlights the global reach and impact of these legacy chemicals decades after their peak use.

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.

Sediment-associated aliphatic and aromatic hydrocarbons in coastal British Columbia, Canada: concentrations, composition, and associated risks to protected sea otters.

Sediment-associated hydrocarbons can pose a risk to wildlife that rely on benthic marine food webs. We measured hydrocarbons in sediments from the habitat of protected sea otters in coastal British Columbia, Canada. Alkane concentrations were dominated by higher odd-chain n-alkanes at all sites, indicating terrestrial plant inputs. While remote sites were dominated by petrogenic polycyclic aromatic hydrocarbons (PAHs), small harbour sites within sea otter habitat and sites from an urban reference area reflected weathered petroleum and biomass and fossil fuel combustion. The partitioning of hydrocarbons between sediments and adjacent food webs provides an important exposure route for sea otters, as they consume ∼25% of their body weight per day in benthic invertebrates. Thus, exceedences of PAH sediment quality guidelines designed to protect aquatic biota at 20% of the sites in sea otter habitat suggest that sea otters are vulnerable to hydrocarbon contamination even in the absence of catastrophic oil spills.

Biota: sediment partitioning of aluminium smelter related PAHs and pulp mill related diterpenes by intertidal clams at Kitimat, British Columbia.

The question of polycyclic aromatic hydrocarbon (PAH) bioavailability and its relationship to specific PAH sources with different PAH binding characteristics is an important one, because bioavailability drives PAH accumulation in biota and ultimately the biochemical responses to the PAH contaminants. The industrial harbour at Kitimat (British Columbia, Canada) provides an ideal location to study the bioavailability and bioaccumulation of sediment hydrocarbons to low trophic level biota. Samples of soft shell clams (Mya arenaria) and intertidal sediment collected from multiple sites over six years at various distances from an aluminium smelter and a pulp and paper mill were analysed for 106 PAHs, plant diterpenes and other aromatic fraction hydrocarbons. Interpretation using PAH source ratios and multivariate data analysis reveals six principal hydrocarbon sources: PAHs in coke, pitch and emissions from anode combustion from the aluminium smelter, vascular plant terpenes and aromatised terpenes from the pulp and paper mill, petroleum PAHs from shipping and other anthropogenic activities and PAHs from natural plant detritus. Harbour sediments predominantly contain either pitch or pyrogenic PAHs from the smelter, while clams predominantly contain plant derived PAHs and diterpenes from the adjacent pulp mill. PAHs from the smelter have low bioavailability to clams (Biota-Sediment Accumulation Factors; BSAFs <1 for pitch and coke; <10 for anode combustion, decreasing to ∼0.1 for the mass 300 and 302 PAHs), possibly due to binding to pitch or soot carbon matrices. Decreases in PAH isomer ratios between sediments and clams likely reflect a combination of variation in uptake kinetics of petroleum PAHs and compound specific metabolism, with the importance of petroleum PAHs decreasing with increasing molecular weight. Plant derived compounds exhibit little natural bioaccumulation at reference sites, but unsaturated and aromatised diterpenes released from resins by industrial pulping processes are readily accumulated by the clams (BSAFs >500). Thus while most of the smelter associated PAHs in sediments may not be bioavailable to benthic organisms, the plant terpenes (including retene, totarol, ferruginol, manool, dehydroabietane and other plant terpenes that form the chemical defence mechanism of conifers) released by pulp mills are bioavailable and possess demonstrated toxic properties. The large scale release of plant terpenes by some of the many pulp mills located in British Columbia and elsewhere represents a largely undocumented risk to aquatic biota.

Environmental fractionation of PCBs and PBDEs during particle transport as recorded by sediments in coastal waters.

The Strait of Georgia (British Columbia, Canada) is a hydrologically complex inland sea with a rich abundance and diversity of species of aquatic life. Marine sediments, as both a sink for hydrophobic contaminants and a potential source for aquatic food webs, were collected from 41 sites throughout the 6,900-km(2) Strait of Georgia. The congener-specific concentrations of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs), including BDE-209, were measured. Urban harbors represented hotspots for both PCBs and PBDEs, whereas PBDEs were also found at high concentrations near municipal outfalls. Patterns of PCB distribution were consistent with historical point source emissions in urban areas and environmental distillation toward lighter profiles in remote sites over time. The single congener BDE-209 dominated the PBDEs, accounting for 52% of the average total concentration. However, nonurban deep-water sediment PBDE profiles were both heavier and had higher concentration-weighted average log K(OW) (octanol-water partition coefficient) values compared to shallow samples (percent BDE-209 of total PBDE, 66 versus 32%; log K(OW) , 9.5 versus 8.2, respectively). Collectively, our results suggest that although source signals largely explain PCB and PBDE hotspots in the Strait of Georgia, the combination of physicochemical properties and environmental processes drive divergent compositional fates for the PCBs and the heavier PBDEs in the sediments of the Strait of Georgia.

Do trans-Pacific air masses deliver PBDEs to coastal British Columbia, Canada?

In order to distinguish between 'local' and 'background' sources of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) in coastal British Columbia (Canada) air, we collected samples from two sites: a remote site on western Vancouver Island, and a near-urban site in the Strait of Georgia. Seasonally-integrated samples of vapor, particulate, and rain were collected continuously during 365 days for analysis of 275 PCB and PBDE congeners. While deposition of the legacy PCBs was similar at both sampling sites, deposition of PBDEs at the remote site amounted to 42% (10.4 mg/ha/year) of that at the near-urban site. Additional research into atmospheric circulation in the NE Pacific Ocean will provide more insight into the transport and fate of priority pollutants in this region, but trans-Pacific delivery of PBDEs to the west coast of North America may underlie in part our observations. For example, approximately 40% of >12,000 ten-day back trajectories calculated for the remote site originated over Asia, compared to only 2% over North America.

Persistent organic pollutants (POPs) in killer whales (Orcinus orca) from the Crozet Archipelago, southern Indian Ocean.

Persistent organic pollutants (POPs), including polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and dibenzofurans (PCDFs), are ubiquitous environmental contaminants of which significant concentrations are reported in upper trophic level animals. In 1998, we collected blubber biopsy samples (n=11) from killer whales (Orcinus orca) inhabiting the coastal waters around Possession Island, Crozet Archipelago, southern Indian Ocean, for contaminant analyses. Despite inhabiting an isolated region far removed from industrial activities, these killer whales can presently be considered among the most PCB-contaminated cetaceans in the southern hemisphere, with concentrations ranging from 4.4 to 20.5mg/kg lipid weight (lw). PCDD levels ranged from below the detection limit (5 ng/kg) to 77.1 ng/kg lw and PCDF levels from below the detection limit (7 ng/kg) to 36.1 ng/kg lw. Over 70% of our study animals had PCB concentrations which exceeded a 1.3mg/kg PCB threshold established for endocrine disruption and immunotoxicity in free-ranging harbour seals, suggesting that organic contaminants cannot be ruled out as an additional threat to this declining population.

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.

Partitioning of current-use and legacy pesticides in salmon habitat in British Columbia, Canada.

Current regulatory paradigms have favored a shift from persistent pesticides that amplify in aquatic food webs to pesticides with reduced persistence and bioaccumulative potential (low log K(OW)). Although these new generation pesticides preferentially partition away from food web-associated lipids, aquatic biota may nonetheless be exposed to them via other environmental compartments. To characterize pesticide patterns in coho salmon (Oncorhynchus kisutch) habitat, we studied two salmon-bearing watersheds (agricultural and urban) in British Columbia, Canada's Fraser River valley and one in a remote area of the province's central coast. The agricultural and remote sites exhibited pesticide patterns dominated by current-use pesticides, whereas the urban site was largely dominated by legacy organochlorine pesticides. When adjusted to trans-chlordane concentrations across environmental matrices, correlations were observed between water to sediment ratios for the pesticides and their octanol:water partitioning coefficients (log K(OW); r2=0.48, p < 0.0001); between air to water ratios and Henry's Law coefficients (r2=0.55, p<0.0001); and between fish to water ratios and log K(OW) (r2=0.74, p<0.0001). These relationships underscore the importance of physicochemical properties in determining the fate of pesticides in freshwater salmon habitat, and highlight the need for research on the nature of health risks associated with exposure where little or no accumulation occurs.

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.

PCB-related alteration of thyroid hormones and thyroid hormone receptor gene expression in free-ranging harbor seals (Phoca vitulina).

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; p < 0.001] and a concomitant decrease in circulating total thyroxine concentrations (capital sigmaPCBs; r = -0.711; p < 0.001) . 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.

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.