Evgeny A. Petrov

Persistent organochlorine residues and their accumulation kinetics in Baikal seal (Phoca sibirica) from Lake Baikal, Russia.

Organochlorine compounds (OCs) such as DDTs (DDT and its metabolites), PCBs (polychlorinated biphenyls), CHLs (chlordane compounds), and HCHs (hexachlorocyclohexanes) were determined in the blubber of Baikal seal (Phoca sibirica) and their fish diet collected from Lake Baikal in 1992. Residue levels of DDTs and PCBs were in the ranges of 4.9-160 μg/g and 3.5-64 μg/g on a lipid weight basis, respectively. The concentrations of CHLs and HCHs were approximately 1 or 3 orders of magnitude lower than those of DDTs and PCBs. Comparison of OC residue levels with those reported for other pinnipeds suggests that Baikal seal is highly contaminated species vulnerable to OC toxicity. A positive age-dependent accumulation of DDTs, PCBs, and CHLs was found in males, while a steady state observed in females suggested the transfer of these chemicals from mother to pup through gestation and lactation. On the basis of contaminant burdens in adult seals, it was estimated that an adult female Baikal seal transfers about 20% of its total DDTs and 14% of its total PCBs to the pup during a reproductive process. Based on the data from isomer-specific analysis of PCBs, it can be suggested that Baikal seals have a higher or comparable capacity to metabolize toxic contaminants than marine mammals, but it is apparently lower than terrestrial mammals, which seems to be a causative factor for the higher accumulation of OC residues in this species.

Comparison of trace element accumulation in Baikal seals (Pusa sibirica), Caspian seals (Pusa caspica) and northern fur seals (Callorhinus ursinus)

Concentrations of 18 trace elements (V, Cr, Mn, Co, Cu, Zn, Se, Rb, Sr, Zr, Mo, Ag, Cd, Sb, Cs, Hg, Tl and Pb) were determined in liver, kidney, muscle and hair of Baikal seals, Caspian seals and northern fur seals. All the three species showed the highest concentrations of Hg, V, Mn, Se and Ag in liver, Cd, Co and Tl in kidney, and Cs in muscle among the soft tissues examined. The highest burdens of Zn, Rb and Cs were observed in muscle, Mo and Ag in liver, and Sb and Pb in hair in all the three species. Concentrations of non-essential elements, Rb, Cd, Cs and Hg, showed significant positive correlations among liver, kidney and muscle, whereas correlation coefficients for essential elements, Mn, Co, Cu, Zn and Se, between the three tissues were generally low for all the species, suggesting that homeostasis controls the concentrations of essential elements but not the non-essential elements in the tissues of these animals. Significant age-dependent increase was found in the concentrations of V, Se and Ag in liver and Hg in liver and kidney of all the three species. Hair concentrations showed significant positive correlations with Zn levels in liver and kidney and Hg in muscle for Caspian seals, Hg in liver and kidney for Baikal seals, and Pb in liver for northern fur seals. Furthermore, regression analysis using the data in the present study and in the literature showed significant positive correlations between Hg levels in hair, and liver, kidney and muscle for various species of pinnipeds. These results indicate the possibility of using hair samples for monitoring these trace elements in pinnipeds.

Toxicokinetics of PCDD, PCDF, and coplanar PCB congeners in Baikal seals, Pusa sibirica: age-related accumulation, maternal transfer, and hepatic sequestration.

To assess the toxicokinetic behavior and potential toxicity of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and coplanar polychlorinated biphenyls (PCBs) in Baikal seals, congener-specific levels and tissue distribution were evaluated in the liver and blubber, and the effects of biological factors including sex and growth were assessed. Total 2,3,7,8-TCDD toxic equivalents (TEQs) were in the range of 210-920 pgTEQ/g fat wt (180-800 pgTEQ/g wet wt) in the blubber and 290-7800 pgTEQ/g fat wt (10-570 pgTEQ/wet wt) in the liver. Non-ortho coplanar PCB126 was the most TEQ-contributed congener accounting for 37-59% of the total TEQs in the liver. From the unique congener profiles, weak metabolic properties of Baikal seals for 2,3,7,8-TCDF and 1,2,3,7,8-P5CDF are suggested. Concentrations of most congeners linearly increased with age in male seals, whereas in adult females the levels revealed an age-related decline. The increasing and declining rates were congener-specific. Maternal transfer rates of 5 representative congeners from adult female to pup through lactation, which was estimated from male-female differences in the body burden, was 1.1 ngTEQ/kg/day for the first pup and decreased with every lactational epoch. The liver-blubber distribution of 1,2,3,4,7,8-H6CDD, 1,2,3,6,7,8-H6CDD, PCB81, PCB126, and PCB169 was dependent on the hepatic total TEQ, indicating hepatic sequestration by induced cytochrome P450 (CYP). These results indicate that congener profile in Baikal seals is governed by complex factors including sex, tissue concentration, binding to CYP, and rates of absorption and metabolism/excretion.

Bioaccumulation profiles of polychlorinated biphenyls including coplanar congeners and possible toxicological implications in Baikal seal (Phoca sibirica).

Isomer specific concentrations of individual polychlorinated biphenyls (PCBs) including toxic non-ortho (IUPAC 77, 126, 169), mono-ortho (105, 118, 156) and di-ortho (137, 138, 153, 180) coplanar congeners were determined in the blubber of 40 Baikal seals (Phoca sibirica) and as their fish diet collected from Lake Baikal, Siberia. Residue levels of total PCBs in Baikal seals were noticeably high and comparable to those reported for seals from the North Sea, suggesting the recent usage of this compound in the watershed of Lake Baikal. Non-, mono-, and di-ortho coplanar congeners were also detected in Baikal seals and fish. An approach to estimate bioaccumulation profiles of PCB congeners revealed that the non-ortho PCBs, IUPAC 77, 126 and 169 seemed to be less persistent than other congeners. Furthermore, selective biotransformation of PCB congeners having either meta-para vicinal H atoms or both adjacent chlorinated meta-para and ortho-meta positions has been suggested. Comparison of 2,3,7,8-TCDD toxic equivalents (TEQ) of non-, mono- and di-ortho coplanar congeners in Baikal seals with those for other marine mammals suggested higher enrichment of mono-ortho congeners, particularly IUPAC 105 and 118, which contributed significantly to the total TEQs in Baikal seals. Results imply that the TCDD-like toxicity is relatively serious in Baikal seals, because of the enrichment of these toxic PCB congeners in tissues.

Trace element accumulation in Baikal seal (Phoca sibirica) from the Lake Baikal.

Trace element concentrations (Fe, Mn, Zn, Cu, Pb, Ni, Cd, Co and Hg) were determined in 60 Baikal seals and in fishes collected from Lake Baikal in 1992. Low levels of Hg and Cd were found in Baikal seals in comparison with those of marine mammals and it was due to their low concentrations in dietary fish. These results suggest that pollution by Hg and Cd was low in Lake Baikal and these toxic elements were unlikely to be the causative factors for mass mortality of Baikal seal in 1987-1988. Significant correlation of Hg concentration between hair and internal tissues suggested the use of hair for Hg monitoring in pinnipeds. Among essential elements, higher Fe and lower Cu levels were specifically found in the liver of Baikal seal. The noticeable accumulation of essential elements might be related to the unique and specific environment of Lake Baikal.

Accumulation features and temporal trends of PCDDs, PCDFs and PCBs in Baikal seals (Pusa sibirica).

This study investigated the accumulation features and temporal trends of PCDD/Fs, dioxin-like PCBs (DL-PCBs) and non-dioxin-like PCBs (NDL-PCBs) in the blubber of Baikal seals collected in 1992 and 2005. DL-PCBs (480-3600ng/g) and NDL-PCBs (980-35,000ng/g) were dominant contaminants. Concentrations of PCDDs and PCBs in males were significantly higher than in females. In males, age-dependent accumulation was observed for PCDDs, mono-ortho PCBs and NDL-PCBs. PCDFs and non-ortho PCBs showed no such trends, implying that exposure of seals to these contaminants has been decreasing in recent years. No decreasing temporal trend was observed for PCDDs, mono-ortho PCBs and NDL-PCBs, suggesting that Baikal seals are still exposed to PCDDs and PCBs. TEQs of PCDDs and mono-ortho PCBs in seals collected in 2005 accounted for 62-77% of total TEQs. The TEQ levels in 40% of the specimens exceeded the threshold level for immunosuppression observed in harbor seals (209pg/g).

Contamination of Tris(4-Chlorophenyl) Methane and Tris(4-Chlorophenyl) Methanol in Marine Mammals from Russia and Japan: Body Distribution, Bioaccumulation and Contamination Status

Four seal species (Phoca caspica, Phoca sibirica, Phoca hispida and Phoca largha) and one whale (Orcinus orca) from Russia and Japan were examined to determine the body distribution, bioaccumulation and contamination status of tris(4-chlorophenyl) methane (TCPMe) and tris(4-chlorophenyl) methanol (TCPMOH). Lipid normalized concentrations of TCPMe and TCPMOH were comparable in various organs and tissues, implying that their body distribution is followed to the lipid-dependent accumulation, similar to that for other organochlorines. The highest body burden of these compounds was found in the blubber. Bioaccumulation potential of TCPMe and TCPMOH was high and comparable to PCBs and DDTs. Relatively higher concentrations of TCPMe and TCPMOH were observed in Caspian seal than in other seal species examined. TCPMOH concentration in killer whale was the highest of the marine mammals examined. Another peak detected had similar mass spectrum to TCPMOH, however, at a different retention time, suggesting the presence of an isomer of TCPMOH.

Integrative assessment of potential effects of dioxins and related compounds in wild Baikal seals (Pusa sibirica): application of microarray and biochemical analyses.

We have previously indicated that accumulation of chlorinated dioxins and related compounds (DRCs) induced cytochrome P450 (CYP) 1A1, 1A2 and 1B1 isozymes in the liver of wild Baikal seals (Pusa sibirica). Here we attempt to assess the potential effects of DRCs triggered by the induction of these CYP1 isozymes in this species, using an integrative approach, combining gene expression monitoring and biochemical assays. To screen genes that may potentially respond to the exposure of DRCs, we constructed a custom cDNA oligo array that can target mRNAs in Baikal seals, and monitored hepatic mRNA expression levels in the wild population. Correlation analyses between the hepatic total 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalents (TEQs) and mRNA levels supported our previous findings that high accumulation of DRCs induces the transcription of CYP1A1, CYP1A2 and CYP1B1 genes. In addition, our integrative assessment indicated that the chronic exposure to DRCs may alter the hepatic transcript levels of genes related to oxidative stress, Fe ion homeostasis, and inflammatory responses. The expression levels of CYP1A2 showed significant positive correlations with levels of malondialdehyde, a biomarker of lipid peroxidation, and of etheno-dA, a DNA adduct, suggesting that the lipid peroxidation may be enhanced through the production of reactive oxygen species (ROS) triggered by CYP1A2 induction. Moreover, there was a positive correlation between heme oxygenase activities and malondialdehyde levels, suggesting the prompted heme degradation by ROS. Fetuin-A levels, which are suppressed by inflammation, showed a significant negative correlation with TEQ levels, and hepcidin levels, which are conversely increased by inflammation, had significant positive correlations with malondialdehyde and etheno-dA levels, implying the progression of inflammation by DRC-induced oxidative stress. Taken together, we propose here that wild Baikal seals may suffer from effects of chronic exposure to DRCs on the induction of CYP1 isozymes, followed by increased oxidative stress, heme degradation and inflammation.

Radionuclide (137Cs and 40K) concentrations in the muscle of Baikal seal (Pusa sibirica) from Lake Baikal

a Ehime Prefectual Institute of Public Health and Environmental Science, Sanban-cho 8-234, Matsuyama, Ehime 790-0003, Japan b Center for Marine Environmental Studies (CMES), Ehime University, Bunkyo-cho 2-5, Matsuyama, Ehime 790-8577, Japan c Thermo Fisher Scientific K.K., Moriya-cho 3-9 C-2F, Kanagawa-ku, Yokohama, Kanagawa 221-0022, Japan d Ehime Prefectual Institute of Uwajima Health Center, Tenjin-cho 7-1, Uwajima, Ehime 798-8511, Japan e Baikal Institute of Nature Management, Siberian Branch of Russian Academy of Sciences, Ulan-Ude, Buryatia 670047, Russia f The Eastern-Siberian Scientific and Production Fisheries Center, ‘‘VOSTSIBRYBCENTR”, Ulan-Ude, Buryatia 670034, Russia

Evaluation of relative potencies for in vitro transactivation of the baikal seal aryl hydrocarbon receptor by dioxin-like compounds.

To evaluate the sensitivity and responses to dioxins and related compounds (DRCs) via aryl hydrocarbon receptor (AHR) in Baikal seals (Pusa sibirica), we constructed an in vitro reporter gene assay system. Baikal seal AHR (BS AHR) expression plasmid and a reporter plasmid containing CYP1A1 promoter were transfected in COS-7 cells. The cells were treated with six representative congeners, and dose-dependent responses were obtained for all the congeners. EC50 values of 2,3,7,8-TCDD, 1,2,3,7,8-PeCDD, 2,3,7,8-TCDF, 2,3,4,7,8-PeCDF, and PCB126 were found to be 0.021, 1.8, 0.16, 2.4, and 2.5 nM, respectively. As the response did not reach the maximal plateau, EC50 value for PCB118 could not be obtained. The TCDD-EC50 for BS AHR was as high as that for dioxin sensitive C57BL/6 mouse AHR. The in vitro dose responses were further analyzed following an established systematic framework and multiple (20, 50, and 80%) relative potencies (REPs) to the maximum TCDD response. The estimates revealed lower REP ranges (20-80%) of PeCDD and PeCDF for BS AHR than for mouse AHR. Average of the 20, 50, and 80% REPs was designated as Baikal seal specific TCDD induction equivalency factor (BS IEF). The BS IEFs of PeCDD, TCDF, PeCDF, PCB126, and PCB118 were estimated as 0.010, 0.018, 0.0078, 0.0059, and 0.00010, respectively. Total TCDD induction equivalents (IEQs) that were calculated using BS IEFs and hepatic concentrations in wild Baikal seals corresponded to only 12-31% of 2005 WHO TEF-derived TEQs. Nevertheless, about 50% of Baikal seals accumulated IEQs over the TCDD-EC50 obtained in this study. This assessment was supported by the enhanced CYP1A1 mRNA expression found in 50% of the specimens contaminated over the TCDD-EC50. These findings suggest that the IEFs proposed from this in vitro assay could be used to predict AHR-mediated responses in wild seals.