Autometallographic tracing of mercury in frog liver

The distribution of mercury in the liver of the frog Rana ridibunda with the autometallographic method was investigated. The mercury specific autometallographic (HgS/SeAMG) technique is a sensitive histochemical approach for tracing mercury in tissues from mercury-exposed organisms. Mercury accumulates in vivo as mercury sulphur/mercury selenium nanocrystals that can be silver-enhanced. Thus, only a fraction of the Hg can be visualized. Six animals were exposed for one day and another group of six animals for 6 days in 1 ppm mercury (as HgCI2 ) dissolved in fresh water. A third group of six animals, served as controls, were sacrificed the day of arrival at the laboratory. First, mercury appears in the blood plasma and erythrocytes. Next, mercury moves to hepatocytes and in the apical part of the cells, that facing bile canaliculi. In a next step, mercury appears in the endothelial and Kupffer cells. It seems likely that, the mercury of hepatocytes moves through bile canaliculi to the gut, most probably bound to glutathione and/or other similar ligands. Most probably, the endothelial and Kupffer cells comprise the first line of defense against metal toxicity.     

Validation of oxidative stress responses in two populations of frogs from Western Ukraine

Although amphibians are considered to be good bioindicators of environmental pollution, few data are available concerning their biochemical parameters in natural populations. We investigated seasonal (spring, summer, autumn) and spatial (wetlands in rural and urban areas) fluctuations of oxidative stress biomarkers in the liver of frog Rana ridibunda in Western Ukraine. The Centroid grouping analysis demonstrated that despite the fluctuations of separate indices, frogs from an urban site in summer and autumn are differed widely from those at the same site in spring and frogs from the rural site in all three seasons, joined in common set. In summer, suppression of Mn-superoxide dismutase and catalase activity, as well as increase in oxidized glutathione and lipid peroxidation levels demonstrate a collapse of antioxidant defense system in frogs from an urban site. The integrated oxidative stress index confirms this conclusion.     

Heavy metal accumulation and metallothionein concentration in the frog Rana ridibunda after exposure to chromium or a mixture of chromium and cadmium

The accumulation of two heavy metals (chromium (Cr) and cadmium (Cd)) in the liver, kidney and gut of Rana ridibunda exposed to Cr or to a mixture of Cr and Cd was investigated. The concentration of metallothioneins (MTs) in the same tissues was also studied. Both metals accumulated mainly in the kidney. Cr accumulation in the liver and gut was not affected by the presence of Cd. Furthermore, Cr concentration in the kidney was doubled when Cd was present. MT concentration did not increase after Cr treatment but it increased two- to six-fold over control values in mixture-exposed frogs, the highest value being observed in the gut. MTs in the gut could act as a barrier preventing ingested heavy metals from entering the blood stream. MT concentration correlated positively with Cd concentrations in both the liver and the gut of mixture-exposed animals.