HEAVY METAL INDUCED TOXICITY IN THE INDIAN GREEN FROG: BIOCHEMICAL AND HISTOPATHOLOGICAL ALTERATIONS

Abstract

Heavy metal contamination may have adverse effects on wetland biota, particularly on amphibians. Severe immunotoxic effects elicited in Euphlyctis hexadactylus (Indian green frog) because of metal exposure (Cd, Cr, Cu, Pb, and Zn) in the Bellanwila-Attidiya Sanctuary, a polluted urban wetland in Sri Lanka, provided the rationale for the present study. We evaluated the biochemical and histopathological effects of this metal contamination with a reference E. hexadactylus population and a laboratory exposure group that was subjected to 28 d of exposure to a mixture of Cd, Cr, Cu, Pb, and Zn (5 ppm in each mixture). A histopathological scoring for the semiquantification of tissue damage was established. Results of the biochemical and histopathological markers were remarkably consistent between the 2 exposure scenarios, providing validation for the heavy metal exposure hypothesis. Damage to liver, kidney, lung, and skin of metal-exposed E. hexadactylus quantified multiple impairments absent in the reference frogs. Liver injuries complemented significantly elevated aspartate transaminase (AST), alanine transaminase (ALT), γ-glutamyl transferase (γ-GT), and alkaline phosphatases in frog liver homogenate, indicating hepatocellular leakage and loss of functional and structural integrity of the hepatocyte membrane in both field- and laboratory-exposed frogs. Significant elevation of Kupffer cell hypertrophy, pigmentation, inflammatory cell infiltrates and hepatic inflammation, extramedullary hematopoiesis, karyocytomegaly of hepatocytes (p < 0.05) of the liver, and degeneration of epithelia and necrosis of the lung, manifested as impairments in both metal exposure scenarios. Significantly reduced serum total protein and albumin and significantly elevated urea and creatinine in metal-exposed frogs were indicative of hepatic and renal dysfunction, respectively. The present study affirms histopathology-related biochemical alterations as potential biomarkers for heavy metal toxicity in amphibians. Environ Toxicol Chem 2017;36:2855–2867. © 2017 SETAC