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.

Association of heavy metals with metallothionein and other proteins in hepatic cytosol of marine mammals and seabirds.

Distribution of Cu, Zn, Cd, Ag, Hg, and Se were determined in hepatocytosol of northern fur seals (Callorhinus ursinus), black-footed albatrosses (Diomedea nigripes), and Dall's porpoises (Phocoenoides dalli). Copper, Zn, and Cd were accumulated preferentially in metallothionein (MT) fraction and their contents in MT fraction increased with the amounts in the hepatocytosol. Silver was bound to both high-molecular-weight substances (HMWS) and MT in the hepatocytosol for all three species, whereas the distribution of Ag in the cytosol was different among the three species. In northern fur seals, Ag mainly was bound to MT, whereas it mainly was associated with HMWS in Dall's porpoises. In contrast, Ag was distributed almost equally in both HMWS and MT for black-footed albatrosses. Mercury content in HMWS and Se content in HMWS and low-molecular-weight substances (LMWS) increased with their contents in hepatocytosol for all the three species. A significant positive correlation was found between Se and Hg contents in high-molecular weight (HMW) fraction in cytosol. The molar ratio of Hg and Se was close to unity in HMW fraction of the specimens with high Hg concentration in cytosol, implying that the Hg-Se complex was bound to the HMWS. Analysis of metals in the hepatocytosol by high-performance liquid chromatography/inductively coupled plasma-mass spectrometry (HPLC/ICP-MS) suggests that multiple isoforms of MT are present in hepatocytosol of the three species and that the metal profiles in hepatocytosols are different among the species. To our knowledge, this is the first report on the association of Ag with HMWS and MT in hepatocytosol of marine mammals and seabirds. Also, distribution and interaction of Hg and Se were investigated for the first time in hepatocytosol of the higher trophic marine animals.

Identification of constitutive androstane receptor cDNA in northern fur seal (Callorhinus ursinus).

Constitutive androstane receptor (CAR) plays a key role in the transcriptional regulation of CYP2B, 2C and 3A genes in response to phenobarbital, ortho-chlorine substituted polychlorinated biphenyls (PCBs) and sex steroids in rodents and human. However, studies addressing CAR are limited to certain laboratory animals and cell lines, and there is little information on the presence of CAR and its physiological and contaminant-related functions in wildlife. While aquatic mammals including seal species are at the top of food chain and highly contaminated by xenochemicals such as PCBs, induction of CYP2/3 subfamilies by such chemical exposure and their regulatory mechanisms have not yet been established in these animals. To investigate mechanisms of CAR-CYPs signaling pathways in aquatic mammals, we initially attempted to isolate CAR cDNA in the liver of northern fur seals (Callorhinus ursinus) from off-Sanriku, Japan. The full-length CAR cDNA had an open reading frame of 1047 bp that encodes a protein containing 348 amino acids. Comparison of the amino acid sequence of CAR from the fur seal with those from other mammalian species showed high identities with CARs from human (83%), monkey (82%), rat (76%) and mouse (73%), revealing a conservation of CAR among the mammalian species. Phylogenetic analysis demonstrated that the fur seal CAR was classified into CAR clade and not into PXR/BXR or VDR clade, suggesting the CARs would be conserved among divergent mammals including aquatic species. With our concomitant paper, where CAR cDNA isolation from the liver of Baikal seal is reported (Iwata et al., in preparation), to our knowledge, this is the first study on the identification of CAR cDNA from wildlife species.

Polybrominated diphenyl ethers and organochlorines in archived northern fur seal samples from the Pacific coast of Japan, 1972-1998.

The present study clearly indicated possible prolonged and chronic marine pollution by polybrominated diphenyl ethers (PBDEs) unless the international regulatory measures are reinforced substantially. Fat tissues of female northern fur seals (Callorhinus ursinus), collected at 10 time periods between 1972 and 1998 from the Pacific coast of northern Japan, were analyzed for PBDEs and organochlorine compounds (OCs). PCBs and DDTs were the predominant contaminants in the fur seal samples. PBDEs were detected in all the samples analyzed, the values ranging from 0.33 to 100 ng/g lipid wt. with predominance of BDE-47. The lowest PBDE levels were in the fur seals collected in 1972, with the peak concentration around 1991-1994 and then decreased to about 50% in 1997-1998. Compositions of higher brominated congeners increased since 1972, while some lower brominated congeners decreased, implying a change in the pattern of use, particularly the increased use of highly brominated diphenyl ethers during 1972-1998. In the meantime, PCB compositions in fur seals showed no temporal variation, suggesting a continuous input of PCBs into the marine environment in significant quantities. As peak concentrations of PBDEs occurred later than OCs, it is essential to follow up the patterns of PBDEs pollution that may be of great concern in the future. To our knowledge, this is the first report on the contamination by PBDEs in marine mammals from the Asia-Pacific waters.