Influence of dissolved oxygen conditions on toxicity of ammonium nitrate to larval natterjack toads

Temporary ponds, where many amphibians from temperate regions breed, show an annual cycle with a maximum water volume in spring followed by a progressive desiccation throughout late spring and summer. This desiccation leads to a decrease in dissolved oxygen and an increase in nitrogen levels, which can additionally increase because of anthropogenic sources such as chemical fertilizers. We analyzed the toxicity posed by environmentally relevant levels of a common nitrogenous fertilizer, ammonium nitrate, at different conditions of oxygen availability to Bufo calamita tadpoles, which typically develop in ephemeral ponds. Ammonium nitrate (90.3 mg N-NO3NH4/l) and hypoxic conditions (initial dissolved oxygen 4.53 ± 0.40 mg/l) caused significant lethal effects after 7 and 12 days of exposure, respectively. At the end of experiment (16 days), mortality rates were 32.5 % in individuals exposed to the fertilizer and 15 % in those growing under hypoxic conditions. When both stressors were combined, they showed an additive effect on tadpole survival. Malformations, such as oedemas and spinal curvatures, and locomotory abnormalities, were detected after 12 days of experiment in >90 % of individuals exposed to 45.2 mg N-NO3NH4/l under hypoxic conditions, whereas none of these stressors by separate related to abnormality rates >35 %. Delayed development was also observed in tadpoles exposed to ammonium nitrate with hypoxia affecting developmental rate only after 12 days of exposure. The results are discussed in terms of potential mechanisms linking negative effects of both factors as well as in terms of potential alterations of the ecological plasticity that often allows amphibians to survive in unpredictable environments.