Altered development, oxidative stress and DNA damage in Leptodactylus chaquensis (Anura: Leptodactylidae) larvae exposed to poultry litter

Abstract

Poultry litter (PL), which is usually used as organic fertilizer, is a source of nutrients, metals, veterinary pharmaceuticals and bacterial pathogens, which, through runoff, may end up in the nearest aquatic ecosystems. In this study, Leptodactylus chaquensis at different development stages (eggs, larval stages 28 and 31 here referred to as stages I, II and III respectively) were exposed to PL test sediments as follows: 6.25% (T1), 12.5% (T2); 25% (T3); 50% (T4); 75% (T5); 100% PL (T6) and to dechlorinated water as control. Larval survival, development endpoints (growth rate -GR-, development rate -DR-, abnormalities), antioxidant enzyme activities (Catalase -CAT- and Glutathione-S-Transferase -GST-), and genotoxic effect (DNA damage index by the Comet assay) were analyzed at different times. In stage I, no egg eclosion was observed in treatments T3-T6, and 50% of embryo mortality was recorded after 24 h of exposure to T2. In stages II and III, mortality in treatments T3-T6 reached 100% between 24 and 48 h. In the three development stages evaluated, the DR and GR were higher in controls than in PL treatments (T1, T2), except for those T1-treated larvae of stage II. Larvae of stage I showed five types of morphological abnormalities, being diamond body shape and lateral displacement of the intestine the most prevalent in T1, whereas larvae of stages II and III presented lower prevalence of abnormalities. In stage I, CAT activity was similar to that of control (p>0.05), whereas it was higher in T1- and T2- treated larvae of stages II and III than controls (p<0.05). In stages I and III, GST activity was similar to that of controls (p>0.05), whereas it was inhibited in T1-treated larvae of stage II (p<0.05). T1- and T2-treated larvae of stages II and III caused higher DNA damage respect to controls (p<0.05), varying from medium to severe damage (comet types II, III and IV). These results showed that PL treatments altered development and growth and induced oxidative stress and DNA damage, resulting ecotoxic for L. chaquensis larvae.