Cardiotoxicity in African clawed frog (Xenopus laevis) sub-chronically exposed to environmentally relevant atrazine concentrations: Implications for species survival

The toxic effects of different atrazine concentrations on tadpoles and adult male African clawed frogs (Xenopus laevis) were assessed in a controlled laboratory environment following 90 days' exposure. The aim was to elucidate the danger of atrazine exposure on the cardiac tissue relative to its critical function of rhythmic contractility, fundamental for optimal blood circulation and homeostasis. Tadpoles and adult frogs were exposed to 0 mu g/L (control), 0.01 mu g L-1, 200 mu g L-1 and 500 mu g L-1 concentrations of atrazine for 90 days. Mortality was concenration-dependent and significantly increased in juvenile group (77%, 43%, 23% and 0 respectively for 500 mu g L-1, 200 mu g L-1, 0.01 mu g L-1, and control group). While the mean juvenile heart area decreased concentration-dependently, adult frog mean heart area significantly increased in the 200 mu g L-1 group only and mean heart weight change was variable across all exposure levels. Light microscopy of hematoxylin and eosin (H&E) and Mallory-Heidenhain rapid one-step staining techniques on cardiac tissue sections of the juvenile and adult frogs revealed shrinkage of cardiac muscle cells into thin wavy myocytes. Additionally, disorganized branching of muscle fibres with reduced striations were observed in 0.01 mu g L-1 and 200 mu g L-1 but hypertrophied myocytes, thickened intensely staining myofibrils in the 500 mu g L-1 group in juvenile and adult frogs. Significant increase in the mean percentage area of connective tissue in all the treated groups (p < 0.036) were also recorded. Immunohistochemistry analysis showed decreased eNOS localization in cardiac tissue in 200 mu g L-1 and 500 mu g L-1 of both juvenile and adult group, suggestive of decreased cardiac contractility due to atrazine exposure. The results indicate that environmentally relevant atrazine concentrations cause significant mortality in tadpoles while concentrations >= 200 mu g L-1 adversely affect cardiac muscle morphology and may induce functional perturbations in cardiac tissue contractility and consequent dysfunction which generally may have an adverse impact on their survival and longevity.