Izumi Watanabe

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.

Using trace elements in skin to discriminate the populations of minke whales in southern hemisphere

Concentrations of 12 trace elements (V, Cr, Mn, Cu, Zn, Se, Rb, Sr, Cd, Cs, Ba, and Hg) were determined in liver and skin tissues of minke whales from various regions within the Antarctic Ocean. Cd concentrations in livers of southern minke whale were apparently higher than those in cetaceans from other regions, while Hg concentrations were lower. There were significant positive correlations between body length and concentrations of Cd and Hg in the liver. The concentrations of all trace elements in the skin were lower than those in other cetaceans reported previously. Significant positive correlations between liver and skin were found for Cr, Mn, Cu, Zn, Rb, Cd, and Cs, implying that the concentrations of these trace elements in the skin reflect those of internal organs. Large interannual variation of the accumulation pattern of trace elements in the skin was observed for the southern minke whales from Area V. There were significant differences in the skin element concentrations among Areas III, IV, and V, especially for males. Also, discriminant analysis between geographically two different groups collected during 1995/1996 austral summer season, based on the concentrations of trace elements in the skin, allowed for a correct classification of 90% of these minke whales. These results suggest that measurement of trace elements in skin samples could provide valuable information on the status of contamination and possible geographic differences in the accumulation levels in southern minke whales.

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.

Contributions of apoplasmic cadmium accumulation, antioxidative enzymes and induction of phytochelatins in cadmium tolerance of the cadmium-accumulating cultivar of black oat (Avena strigosa Schreb.)

The contributions of cadmium (Cd) accumulation in cell walls, antioxidative enzymes and induction of phytochelatins (PCs) to Cd tolerance were investigated in two distinctive genotypes of black oat (Avena strigosa Schreb.). One cultivar of black oat ‘New oat’ accumulated Cd in the leaves at the highest concentration compared to another black oat cultivar ‘Soil saver’ and other major graminaceous crops. The shoot:root Cd ratio also demonstrated that ‘New oat’ was the high Cd-accumulating cultivar, whereas ‘Soil saver’ was the low Cd-accumulating cultivar. Varied levels of Cd exposure demonstrated the strong Cd tolerance of ‘New oat’. By contrast, low Cd-accumulating cultivar ‘Soil saver’ suffered Cd toxicity such as growth defects and increased lipid peroxidation, even though it accumulated less Cd in shoots than ‘New oat’. Higher activities of ascorbate peroxidase (EC 1.11.1.11) and superoxide dismutase (EC 1. 15. 1. 1) were observed in the leaves of ‘New oat’ than in ‘Soil saver’. No advantage of ‘New oat’ in PCs induction was observed in comparison to Cd-sensitive cultivar ‘Soil saver’, although Cd exposure increased the concentration of total PCs in both cultivars. Higher and increased Cd accumulation in cell wall fraction was observed in shoots of ‘New oat’. On the other hand, in ‘Soil saver’, apoplasmic Cd accumulation showed saturation under higher Cd exposure. Overall, the present results suggest that cell wall Cd accumulation and antioxidative activities function in the tolerance against Cd stress possibly in combination with vacuolar Cd compartmentation.

Trace elements accumulation in waders from Asia

Concentrations of V, Cr, Mn, Co, Cu, Zn, Se, Rb, Sr, Mo, Ag, Cd, Sb, Cs, Ba, Hg, Tl, and Pb were determined in the liver of 9 species of waders including migratory and resident collected from north Vietnam, south India, and the Philippines. Renal Cd concentrations were also determined. Most elements showed no significant differences between genders and among localities. Concentrations of almost all the elements in birds, except for Cr and Se in the kentish plover from Vietnam, were found to be decreased or less varied during the wintering season. Toxic element levels in waders were dependent on the migration distance. Pb levels in dunlins from Vietnam and Cd levels in pintail snipe from Philippines seemed to be in toxicologically critical on their health condition.

Trace element distribution and risk assessment in South Indian surface waterways

The concentrations of 27 major and trace elements are determined in surface water samples collected from 48 sites of diverse waterways in four states (Tamil Nadu, Kerala, Karnataka, and Telangana) of South India. The aims of this study are to identify the element distribution, comparatively assess the pollution risk, and evaluate human health risks related to diverse waterways in the study area. The results indicate that elements such as Cr, Se, As, Fe, and Mn are the major pollutants, as their concentrations exceeded the acceptable national and international water quality standards in several sites of Ennore, Adyar, Cooum, Periyar, and Vrishabhavathi rivers. Furthermore, statistical analysis reveals that the Ennore, Adyar, Cooum, Periyar, and Kaveri river basins are affected by various anthropogenic activities, leading to moderate-to-high pollution by As, Cr, Mn, Fe, and Se. Potential pollution sources are industrial waste, sewage intrusion, paint industry waste, and automobile runoff. Overall, the investigated sites are categorized into three major groups: highly, moderately, and least polluted. Risk on human health by metals is then evaluated using hazard quotients (HQs) and carcinogenic risk evaluation; the results indicated that As with HQ >1 is the most hazardous pollutant, which could lead to non-carcinogenic and carcinogenic concerns, particularly in children. This study helps in establishing pollutant loading reduction goal and the total maximum daily loads and consequently contributes to preserving public health and developing water conservation strategies.

Establishment of a primary hepatocyte culture from the small Indian mongoose (Herpestes auropunctatus) and distribution of mercury in liver tissue

The present study established a primary hepatocyte culture for the small Indian mongoose (Herpestes auropunctatus). To determine the suitable medium for growing the primary hepatic cells of this species, we compared the condition of cells cultured in three media that are frequently used for mammalian cell culture: Dulbecco’s Modified Eagle’s Medium, RPMI-1640, and William’s E. Of these, William’s E medium was best suited for culturing the hepatic cells of this species. Using periodic acid-Schiff staining and ultrastructural observations, we demonstrated the cells collected from mongoose livers were hepatocytes. To evaluate the distribution of mercury (Hg) in the liver tissue, we carried out autometallography staining. Most of the Hg compounds were found in the central region of hepatic lobules. Smooth endoplasmic reticulum, which plays a role inxenobiotic metabolism, lipid/cholesterol metabolism, and the digestion and detoxification of lipophilic substances is grown in this area. This suggested that Hg colocalized with smooth endoplasmic reticulum. The results of the present study could be useful to identify the detoxification systems of wildlife with high Hg content in the body, and to evaluate the susceptibility of wildlife to Hg toxicity.

Kinematic gait analysis and lactation performance in dairy cows fed a diet supplemented with zinc, manganese, copper and cobalt.

This study investigated how supplementation of the diet of dairy cows with trace minerals (zinc, manganese, copper and cobalt) affected kinematic gait parameters and lactation performance. Eight Holstein cows were divided into two groups, with each group receiving a different dietary treatment (control diet, or control diet supplemented with trace minerals) in a two-period crossover design. Kinematic gait parameters were calculated by using image analysis software. Compared to cows fed the control diet, cows that received the trace mineral-supplemented diet exhibited significantly increased walking and stepping rates, and had a shorter stance duration. Feed intake and milk production increased in cows fed the trace mineral-supplemented diet compared with control groups. The plasma manganese concentration was not different in control and experimental cows. In contrast, cobalt was only detected in the plasma of cows fed the supplemented diet. These results provide the first evidence that trace mineral supplementation of the diet of dairy cows affects locomotion, and that the associated gait changes can be detected by using kinematic gait analysis. Moreover, trace mineral supplementation improved milk production and only minimally altered blood and physiological parameters in dairy cows.

Subcellular distribution and potential detoxification mechanisms of mercury in the liver of the javan mongoose (Herpestes javanicus) in amamioshima Island, Japan†

In a previous study, we showed that Hg accumulated to high levels in the liver of the Javan mongoose (Herpestes javanicus), a terrestrial mammal that lives on Amamioshima Island, Japan. This suggests a sophisticated mechanism of hepatic Hg detoxication. Assay of the subcellular localization of Hg and the expression of protective enzymes provides important clues for elucidating the mechanism of Hg detoxication. In the present study, the concentrations of 11 elements (Mg, Cr, Mn, Fe, Cu, Zn, Se, Rb, Cd, total Hg [T-Hg] and organic Hg [O-Hg], and Pb) were determined in the liver and in five liver subcellular fractions (plasma membrane, mitochondria, nuclei, microsome, and cytosol) of this species. As the T-Hg level increased, T-Hg markedly distributed to the plasma membrane. The T-Hg levels in all subcellular fractions correlated with Se levels. Although the T-Hg level in the microsomal fraction was relatively low, the ratio of O-Hg to T-Hg was significantly lower in the microsomes than in the other fractions. Significant positive correlations were found between the level of glutathione-S-transferase-π, a marker of oxidative stress, and the O-Hg and T-Hg levels, but the correlation was better with O-Hg than with T-Hg. Western blot analysis of thioredoxin reductase 2 (TrxR2), a protein involved in protecting cells from mitochondrial oxidative stress, showed that the level of TrxR2 correlated with that of T-Hg. High TrxR2 levels may be one mechanism by which the Javan mongoose attenuates the toxicity of the high Hg levels present in the liver.

Seasonal movement of trace-element discharge in a typical south-Indian suburban community

Four conventional activated sludge sewage treatment plants (STPs) in Southern India were chosen to investigate the occurrence of major ions and trace metals in dissolved fraction of the wastewater. Samples were collected from inlet and outlet of STPs during the pre-monsoon and monsoon seasons. Except for a few elements (Mn, Mo and Ni), the concentrations of the rest increased at the outlet of STPs, suggesting that there is an addition of these elements during the treatment process. The increase in concentrations at the STP outlet ranged from 1.25 times for Mn to 3,254 times for Ag during the pre-monsoon. In the monsoon, the increase ranged from 1.75 (Fe) to 1,510 (Ag). This suggests that there is a substantial anthropogenic input of these elements as they pass through the treatment process. Removal rates of elements in STPs also varied. 59% of Mn was removed during the treatment process during pre-monsoon as compared to 67% removal during monsoon.