Temporal Trends in Polybrominated Diphenylethers (PBDEs) in Blubber of Ringed Seals (Pusa hispida) from Ulukhaktok, NT, Canada Between 1981 and 2015.

Blubber taken from ringed seals (Pusa hispida) during a subsistence hunt at Ulukhaktok, NT (formerly Holman, NWT) at intervals between 2002 and 2015 was analysed for polybrominated diphenyl ether (PBDE) congeners. Results from these analyses were combined with others previously published to yield a data set of 18 tri- to hepta-substituted PBDE congeners in 102 animals sampled over a span of 19 year (females) and 34 year (males). In females, mean total PBDE concentrations increased between 1996 and 2015 by approximately 50%, from 1940 to 2780 pg/g wet wt., although not significantly so (p > 0.05) by one-way ANOVA. In males, concentrations ranged from 376 to 6470 pg/g wet wt. between 1981 and 2015 (p < 0.05). In males, the most rapid increase in PBDE concentrations occurred before 2000, but between 2002 and 2015 mean total PBDE concentrations increased by a further 50%. ANCOVA showed PBDE concentrations in females to be correlated (p < 0.05) with sampling year but not with age or condition (as measured by blubber thickness); in males, PBDE concentrations were strongly correlated (p < 0.01) with year, age and condition. The relative proportions of tetra-bromo- congeners declined weakly in both sexes over the sampling period, with a compensatory increase in penta-bromo-congener distribution. Overall, the results show no evidence yet of a decline in PBDE concentrations in western Arctic ringed seals in response to voluntary and regulated restrictions on PBDE use in the early 2000s.

Trends of persistent organic pollutants in ringed seals (Phoca hispida) from the Canadian Arctic.

Ringed seals (Phoca hispida) have been used as bioindicator species of environmental contamination in Canada since the 1970s. In the present study, seals were harvested during subsistence hunts in four regions of the Canadian Arctic: Beaufort Sea, Arctic Archipelago, Hudson Bay, and coastal Labrador. An extensive suite of persistent organic pollutants (POPs) was determined in seal blubber collected for multiple years between 1972 and 2016. Results from this long-term study indicate geographical differences in the contaminant concentrations in seals and the significant general decrease of most POPs, including polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and related compounds, chlordanes (CHL), and hexachlorocyclohexanes (HCH) over time in ringed seals. The highest decrease rates (up to -9.1%/year for α-HCH) were found in seals from the Hudson Bay region where all chemicals investigated have significantly decreased since 1986. Significant increases in concentrations of hexachlorobenzene (HCB) in seals from Labrador and ß-HCH in Sachs Harbour, NT and Arctic Archipelago were observed. Site-specific and contaminant-specific associations between climate pattern (i.e., Arctic Oscillation, North Atlantic Oscillation, and Pacific/North American pattern) and mean ice-coverage (total, first-year ice, and old-ice) were found at sites with the longest time trend data (i.e., Arviat, Sachs Harbour/Ulukhaktok and Resolute Bay). Overall, results suggest that North American and international regulations have led to the long-term reduction of most POPs in Canadian Arctic ringed seals by reducing emissions from primary sources. However, other sources of legacy compounds (e.g., environmental reservoirs) as well changes in food web composition and structure in relation to climate changes could also be influencing the very slow rates of decline, or stable levels, of contaminants found in seals at some sites. Further work is warranted to discern between co-variation of climate changes and contaminant concentrations and cause-and-effect relationships.

Temporal trends in "legacy" organochlorine contaminants in blubber of ringed seals (Phoca hispida) from Ulukhaktok, NT, Canada between 1972 and 2010.

In blubber of ringed seals (Phoca hispida) from Ulukhaktok, NT, residues of DDT, other organochlorine pesticides, and PCBs declined between 1972 and 2010. The rate of decline varied: concentrations of the DDT-group began to fall after 1981, whereas those of PCBs fell rapidly between 1972 and 1981, and then slowed. Concentrations of cis- and trans-chlordane and of HCB in both sexes, and of cis-nonachlor in males, declined slowly between 1978 and 2010; those of other organochlorine pesticides remained steady. Exponential half-lives of p,p'-DDT and p,p'-DDE in female seals are about 9 and 36 y (corresponding to initial declines of 7.8% and 1.9% per year, respectively) and those of PCB congeners from about 20 to 60 y (declines of 3.2% to <1.5% per year); the more refractory residues may be detectable for centuries to come. Exploratory PCA of PCB congener distribution identified temporal changes apparently not related to molecular structure.

Acute toxic responses of the rockfish (Sebastes schlegeli) to Iranian heavy crude oil: feeding disrupts the biotransformation and innate immune systems.

To clarify the toxic effects of Iranian Heavy Crude Oil (IHCO) from the "Hebei spirit" oil spill, innate immune toxic effects defending on biotransformation pathway have been investigated on fish exposed to IHCO. Juvenile rockfish were exposed to IHCO in gelatin capsules by feeding. The effects on multiple fish biotransformation enzymes (Cytochrome P4501A and glutathione-S-transferase) and the expression level of the several immune response genes, including interleukin-1beta, granulocyte colony-stimulating factor and Cathepsin L, were measured in the liver, spleen and kidney. The tissue-specific expression patterns of these genes demonstrated that the highest expression levels of Cytochrome P4501A, glutathione-S-transferase, interleukin-1beta, granulocyte colony-stimulating factor, interferon stimulated gene 15 and Cathepsin L were found in the liver and that the TNF receptor was high in spleen. The oil-fed fish had significantly higher concentrations of biliary fluorescent metabolites and Cytochrome P4501A expression during the initial stage (12 ∼ 48 h after exposure) than those in the liver and kidney of the sham group. Similarly, the highest mRNA expression levels of interleukin-1beta and granulocyte colony-stimulating factor were detected in the liver at the early stages of exposure (12 h after exposure). Following exposure, the levels of interferon stimulated gene 15 and granulocyte colony-stimulating factor mRNA remained high at 120 h after exposure in the liver but the levels of interleukin-1beta and Cathepsin L gradually decreased to an expression level equal to or less than the sham group. Our data suggest that the innate immune and hepatodetoxification responses in oil-fed fish were induced at the initial stage of exposure to the IHCO at the same time but several immune-related genes decreased to less than that of the sham group after the initial stage of response. Therefore, immune disturbances in fish exposed to IHCO may allow the pathogens, including the infectious diseases, to more easily affect the oil exposed fish.

Hexachlorocyclohexanes (HCH) in ringed seal (Phoca hispida) from Ulukhaktok (Holman), NT: trends from 1978 to 2006.

Trends in alpha-, beta-, and gamma-hexachlorocyclohexane (HCH) concentrations were examined in blubber lipid of ringed seals (Phoca hispida) from Ulukhaktok (Holman), NT (Canada) sampled at intervals between 1978 and 2006. alpha-HCH usually represented approximately 90% of the total HCH isomers. alpha-HCH and gamma-HCH concentrations showed no change over the sampling interval, but beta-HCH concentrations increased significantly, about 8-10-fold in females and 4-5-fold in males. Residue concentrations showed no dependence on age. Concentrations (all data as ng/g lipid, GM (range)) of alpha-HCH were significantly higher (P<0.001 by t-test) in males (217 (93.9-517), n=37) than those in females (138 (40.9-402), n=38). beta-HCH concentrations did not differ between the sexes. Concentrations of gamma-HCH were significantly higher (P<0.05) in males (6.74 (0-46.7)) than in females (4.35 (0-19.0)). Although global emissions of both alpha-HCH and beta-HCH have declined since the early 1980's, the "signal" of HCH emission changes has not yet resulted in a "response" in ringed seal residue concentrations. In the light of our current understanding of the dynamics of HCH in the Arctic, we conclude that any such response may not be detected by retrospective analyses of the sort describe here at least for another decade or so, because of the longevity of the seals.

Polybrominated diphenyl ethers (PBDEs) in seal populations from eastern and western Canada: an assessment of the processes and factors controlling PBDE distribution in seals.

Polybrominated diphenyl ether (PBDE) concentrations were measured in the blubber of five mother-pup pairs of grey seals (Halichoerus grypus) from Sable Island, Nova Scotia (NS) sampled in 1995 and in 20 harbour seals (Phoca vitulina) from British Columbia (BC) sampled in 1991-1992. Concentrations in maternal grey seals from Sable Island averaged 112+/-55.2 ng/g lipid (mean+/-s.d.) and were over twice the concentrations measured in their pups; higher brominated PBDEs showed a distinct decline in PBDEs transfer efficiency in mother-pup pairs as a function of increasing K(o/w). Hepta-BDEs and higher molecular weight (MW) congeners appear not to be transferred efficiently from the grey seals mother to pup during lactation on the basis of lipid/water partitioning (as indicated by ln K(o/w)) and this may be a consequence of molecular size (MW>600 Da) which may limit diffusion and partitioning. PBDE concentrations in blubber from harbour seals from the Strait of Georgia averaged 319+/-132 ng/g lipid (mean+/-s.d., n=13), while those from the more remote and less contaminated Quatsino Sd. averaged 27.8+/-11.8 ng/g (mean+/-s.d., n=7). Tri- to hexa-BDEs were appreciably enriched in Strait of Georgia harbour seals relative to those from Quatsino Sd, suggesting that the former were closer to PBDE sources. The main congeners identified in all samples were BDEs 47, 99, 100, 153 and 154. Data were compared to those in ringed seals sampled from Holman, North West Territories (NWT), in 1996. The Holman samples had a higher proportion of lower bromination congeners, and a lower proportion of higher bromination congeners, than did the Sable Island and BC seal samples, consistent with selection in favour of more volatile compounds during atmospheric transport to the Arctic.

Characterization of cholinesterases in marbled sole, Limanda yokohamae, and their inhibition in vitro by the fungicide iprobenfos.

Cholinesterases (ChEs) have been characterized in marbled sole (Limanda yokohamae) for use as a possible biomarker of pollution exposure. In brain, ChEs existed almost exclusively (>95%) as acetylcholinesterase (AChE) whereas in muscle, about 20-30% of ChE activity was in the form of butyrylcholinesterase (BChE; pseudocholinesterase). Acetylthiocholine and butyrylthiocholine (identified in mammalian studies as diagnostic substrates for AChE and BChE respectively) were hydrolyzed mainly, but not exclusively, by these enzymes. The inhibitors BW284C51 and iso-OMPA (identified in mammalian studies as diagnostic inhibitors of AChE and BChE respectively) were not specific for these enzymes in marbled sole. Brain AChE and muscle AChE and BChE were characterized in terms of their kinetic properties (KM etc.) and optimal conditions (substrate concentration, protein concentration, pH etc.) were established to allow routine assays of ChE activity to proceed under pseudo-first order conditions. The sensitivity of ChEs to a locally significant pesticide, iprobenfos (IBP; kitazin) was established in terms of IC50 concentrations. Brain AChE was relatively insensitive to IBP, but muscle AChE and BChE were sensitive to IBP concentrations in the high nM range. However, ambient IBP concentrations in Korean coastal waters are usually not high enough to cause detectable ChE inhibition in this species.

Protein and gene expression of VTG in response to 4-nonylphenol in rockfish (Sebastes schlegeli).

To investigate the in vivo estrogenic activity of 4-nonylphenol (4-NP) on reproductive function, we have measured the expression level of rockfish vitellogenin (rVTG) mRNA, concentration of plasma VTG and levels of plasma estradiol-17beta(E(2)) and testosterone (T) by radioimmunoassay (RIA). An rVTG mRNA transcript of approximately 4.0 kb was extracted from hepatic tissue for Northern blot analysis, and the effects of 4-NP on rVTG mRNA expression in vivo with both male and female juvenile rockfish were examined. The level of rVTG mRNA expression was increased 48 h after injection with 4-NP of 10 mg/kg body mass and after injection in both male and female rockfish. The level of rVTG mRNA expression was increased 24 h after injection in male rockfish injected with 4-NP of 25 mg/kg body mass, while the level of rVTG mRNA expression was increased 12h after injection in female injected with same dosage. Concentrations of plasma rVTG in female injected with 4-NP of 10 and 25 mg/kg b.w. were increased 72 h after injection and reached 35 and 66 mg/mL, respectively. In male, rVTG concentrations with the dosage of 25 mg/kg body mass were increased 72 h after injection and reached to 14 and 65 mg/mL at 168 h, respectively. Plasma concentrations of T and E(2) in female and male injected with 4-NP were not significantly different between two 4-NP dosages. These results suggest that 4-NP may disrupt the reproductive system of immature rockfish by acting directly on vitellogenesis. Immature females injected with 4-NP were more sensitive than immature males, but VTG in immature males is likely to return to normal conditions slowly. There are obviously large interspecies differences in response of fish to 4-NP.

PCDD/F and PCB concentrations in Arctic ringed seals (Phoca hispida) have not changed between 1981 and 2000.

Concentrations of polychlorinated dibenzo-p-dioxins (PCDD) and dibenzofurans (PCDF) and of non-ortho-, mono-ortho- and di-ortho-substituted polychlorinated biphenyls (NO-CB, MO-CB and DO-CB) were measured in blubber of ringed seals sampled at Holman, NWT, in 1981, 1991, 1996 and 2000. Total PCDD and PCDF concentrations were usually below approx. 10 and 5 pg/g wet wt., respectively, and did not change significantly between 1981 and 2000, although there were sporadic temporal differences in some congeners. Total NO-CB, MO-CB and DO-CB concentrations were below approx. 1 ng/g, 250 ng/g and 1 microg/g wet wt. respectively; none of the total PCB concentrations changed significantly over the sampling period. Neither PCDD nor PCDF concentrations differed between males and females, nor did they increase with age in male samples. MO-CB and DO-CB concentrations increased with age in males, but not in females, and concentrations of total MO-CB and DO-CB were usually lower in females than in males. Changes in the distribution of PCB congeners between the 1980s and the 1990s are consistent with atmospheric transport processes becoming increasingly important in the introduction of PCBs to the Arctic in recent years.

PCDD/F and PCB in harbour seals (Phoca vitulina) from British Columbia: response to exposure to pulp mill effluents.

Blubber of harbour seals (Phoca vitulina) sampled from the Strait of Georgia, BC, in 1991 and 1992 contained higher concentrations of polychlorinated dibenzo-p-dioxins and furans (PCDD/F) than did samples from Quatsino Sound on western Vancouver Island. The higher proportions of 1,2,3,6,7,8-hexachlorodibenzodioxin and 2,3,7,8-tetrachlorodibenzofuran in the Strait of Georgia samples probably reflect contamination arising from the discharge of effluents from bleached kraft mills. Higher concentrations of non-ortho- and mono-ortho substituted polychlorinated biphenyls were also found in the Strait of Georgia samples, probably as a result of general industrial activity in the region. Principal components analysis showed that residues were distributed differently between males and females. Within males, concentrations of PCDD, PCDF and PCBs appeared to increase with age. Toxic equivalents (TEQ) to 2,3,7,8-tetrachlorodibenzodioxin in the Quatsino Sd. samples were close to the working guidelines for edible fish used by Health Canada; TEQ calculated for the Strait of Georgia samples exceeded these guidelines by up to a factor of five.

Hepatic CYP1A levels and EROD activity in English sole: biomonitoring of marine contaminants in Vancouver Harbour.

To assess chemical contaminant stress in the marine environment, ethoxyresorufin-O-deethylase (EROD) activity and cytochrome P450 1A (CYP1A) expression were measured in 88 English Sole (Pleuronectes vetulus) collected during May and June 1999 from four sites in Vancouver Harbour and at an expected reference site outside the harbour. Hepatic microsomes were prepared from the fish and analyzed for total CYP content, EROD activity, and CYP1A protein levels. Hepatic EROD activity and CYP1A protein levels were elevated in fish from two sites in the inner harbour. A comparison with sediment chemistry data showed that fish with increased EROD activity and CYP1A levels came from sites containing relatively high levels of polycyclic aromatic hydrocarbons and polychlorinated biphenyls. Unexpectedly high levels of EROD activity and CYP1A protein were also found in fish from a reference site near Gibsons, in Howe Sound. The elevated EROD activity and CYP1A expression in fish from this site cannot be explained by the chemical analysis data collected.

Trends in organochlorine residue concentrations and burdens in grey seals (Halichoerus grypus) from Sable Is., NS, Canada, between 1974 and 1994.

In blubber from maternal grey seals (Halichoerus grypus) from Sable Is., NS, sigma DDT (p,p'-DDE and p,p'-DDT) concentrations declined from 12 micrograms/g lipid in 1974 to 0.5 microgram/g in 1994. Pup blubber sigma DDT concentrations were about 60% of those of their mothers, and declined at similar rates. In maternal seals, p,p'-DDE increased from 56% sigma DDT in 1974 to 89% in 1994. Polychlorinated biphenyl concentrations did not change between 1976 and 1988, but then declined; the relative proportion of chlorobiphenyl (CB) 101 increased, and those of CBs 170 and 187 decreased between 1985 and 1991. Concentrations of alpha-hexachlorocyclohexane and trans-nonachlor declined after 1988, and of oxychlordane after 1992; concentrations of hexachlorobenzene were constant between 1984 and 1994.

Experimental exposure of juvenile chinook (Oncorhynchus tshawytscha) to bleached kraft mill effluent: hepatic CYP1A induction is correlated with DNA adducts but not with organochlorine residues.

Juvenile chinook salmon (Oncorhynchus tshawytscha) were exposed for 28 days to 0-16% treated effluent from an elemental chlorine free (ECF) bleached kraft pulp mill. Fresh effluent was diluted with river water drawn from upstream of the effluent diffuser. Fish were tested for biochemical responses to identify if the effluent would cause significant effects at concentrations spanning those present in the Fraser River, BC, Canada during winter conditions. Hepatic ethoxyresorufin-O-deethylase (EROD) activity was increased significantly at all effluent concentrations and hepatic cytochrome P450 1A (CYP1A) protein was increased in all but 2% effluent. Hepatic DNA adduct concentrations were increased significantly at 8 and 16% effluent. These data indicate that there is a significant increase in all three responses at concentrations similar to those found in the receiving waters (4%) and that a dose-response relationship exists between BKME concentration and the responses measured. Carcasses contained low (< 0.2 pg g(-1)) concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs). Polychlorinated biphenyls (PCBs) were at higher concentrations, accounting for 77% of the total contaminant burden, expressed as TEQ, but contaminants do not appear to have bioaccumulated in this study. It is likely that the PCB concentrations were due to contaminated commercial fish feed. Bile did not contain detectable levels of polycyclic aromatic hydrocarbons (PAHs) as measured by synchronous fluorescence spectrometry.

Similarity of an Aroclor-based and a full congener-based method in determining total PCBs and a modeling approach to estimate Aroclor speciation from congener-specific PCB data.

Polychlorinated biphenyls (PCBs) have entered the environment in North America as Aroclor technical mixtures. Most methods used for the determination of total PCB levels in environmental samples visually match patterns of sample peaks to those in Aroclor standards. Concern over the accuracy of Aroclor-based measurements on compositionally modified samples coupled with advancements in analytical techniques have led to congener-specific PCB analysis. In this study, the PCB data from 27 tissue samples determined by an Aroclor-based method and a full congener method were compared in terms of total PCB concentration to assess the reliability of this Aroclor technique for total PCB determination. Our data show a strong correlation between the sum of Aroclors and the total PCBs obtained from the full congener determinations. We also developed a model using the compositional data from three Aroclors (1242, 1254, and 1260) to determine the amount of compositional alteration from original Aroclor patterns in environmental samples. Full congener data, from a variety of tissue types and trophic levels, examined using this method showed that compositional modification from original Aroclor patterns increases with trophic level, with the greatest modification observed in seal and killer whale samples. This result agrees both with expectation and with what has been found in other studies. Such techniques, which connect congener-specific PCB data to Aroclor contamination, may prove useful to investigations into environmental and metabolic fate and transfer processes.

Immune function in free-ranging harbor seal (Phoca vitulina) mothers and their pups during lactation.

Immune function in harbor seal mothers and their pups during lactation was studied on Sable Island, Nova Scotia, Canada, during the springs of 1989 and 1990. Methods included total white blood cell and differential counts, a Protein A enzyme linked immunosorbent assay (ELISA) for total immunoglobulin G (IgG) quantification, and functionality testing of lymphocytes in vitro using the T-cell mitogen concanavalin A (ConA). Lymphocyte functionality and total IgG levels were reduced in the mothers at the end of lactation, suggesting a reduction in immune function, possibly as a result of the stress of fasting, or hormonal changes associated with lactation and estrus. By contrast, lymphocyte functionality and total IgG levels in pups were low at birth and higher at the end of lactation. Pups at birth and females late in lactation may therefore be more susceptible to infection by viral and bacterial agents. This study represents the first broad examination of immune function in a free-ranging pinniped population.

Acute toxicity and toxicokinetics of chlorinated diphenyl ethers in trout.

1. The LC50 and the uptake and elimination kinetics of 4-chlorodiphenyl ether (4-CDE), 2,4-dichlorodiphenyl ether (2,4-diCDE), 2,4,4'-trichlorodiphenyl ether (2,4,4'-triCDE) and 2,4,5,4'-tetrachlorodiphenyl ether (2,4,5,4'-tetraCDE) were examined in brook trout (Salvelinus fontinalis). The chlorinated diphenyl ethers (CDE) were administered in the water. 2. The 96 h LC50 of 4-CDE and 2,4-diCDE were 0.73 and 0.66 mg/l, respectively. The LC50 of 2,4,4'-triCDE and 2,4,5,4'-tetraCDE could not be determined since they were greater than the water solubilities of these chemicals. 3. Uptake of the CDE by trout was rapid; the uptake rate ranged between 2.4 and 48.9 micrograms/day. However, CDE uptake by trout did not reach a steady state at the conclusion of a 7 day exposure. In contrast, depuration of the CDE from trout was slow; the depuration half-lives of the CDE ranged between 3.9 and 63 days. 4. The toxicokinetics of the CDE in trout could be described by a one-compartment open model with zero-order absorption and first-order elimination. The CDE were taken up initially into the blood and liver of trout before they were redistributed to adipose tissue and muscle. 5. CDE are significantly bioaccumulated by fish from water; the model-predicted maximum body burdens of trout exposed to 100 micrograms/l CDE in water are 146, 316, 1381, and 930 micrograms/g for 4-CDE, 2,4-CDE, 2,4,4'-triCDE and 2,4,5,4'-tetraCDE, respectively.

Distribution of delta-aminolevulinic acid synthetase and delta-aminolevulinic acid dehydratase in liver and kidney of rainbow trout (Salmo gairdnerii).

1. Activities of delta-aminolevulinic acid synthetase (ALA-S) and delta-aminolevulinic acid dehydratase (ALA-D) in trout liver and kidney were compared with those in the mouse. 2. ALA-S activity (per unit tissue fresh weight) exceeded ALA-D activity in trout liver and kidney. 3. In trout kidney, ALA-S activity slightly exceeded, and ALA-D activity far exceeded, their activities in trout liver. 4. In trout, heme synthesis differs from that in mammals in that appreciable synthesis occurs in the kidney, and in that ALA-S activity is not rate limiting.

Studies on the pharmacokinetics and metabolism of 4-chlorodiphenyl ether in rats.

The metabolic disposition of 14C-labeled 4-chlorodiphenyl ether ([14C]4-CDE) was examined in rats following iv administration of a single dose (850 nmol/kg). [14C]4-CDE decayed rapidly from the blood since no unchanged [14C]4-CDE was detected in the blood beyond 2 hr after [14C]4-CDE administration. The dispositional kinetics of [14C]4-CDE in rats were best described by a two-compartment open pharmacokinetic model. Total radioactivity was excreted slowly from rats; about 41% and 33% of the administered dose were excreted into the urine and feces, respectively, within 1 week after chemical administration. About 5% of the total radioactivity administered to rats was excreted into the bile in 1 hr. The bulk of the radioactivity in the excreta was due to the presence of [14C]4-CDE metabolites. 14C-labeled 4'-hydroxy-4-CDE was the major metabolite and accounted for at least 90% of the radioactivity in the urine. The metabolic conversion of [14C]4-CDE to 14C-labeled 4'-hydroxy-4-CDE was corroborated by in vitro studies with liver microsomes of rats. In addition, [14C]4-CDE was converted by liver microsomes to reactive metabolites which bound irreversibly to microsomal protein. An arene oxide is suggested as the intermediate metabolite in the biotransformation of [14C]4-CDE by rats.