Effects of nitrate on embryos of three amphibian species

Embryos of three aquatic breeding amphibian species, Ambystoma mexicanum, Hyla chrysoscelis, and Rana clamitans, were exposed to increasing levels (0, 5, 10, 30, 60, 100, 300, and 500 mg/L) of nitrate-N (NO(3)-) in laboratory, static-renewal experiments. Lethal effects were recorded from Gosner stage 2 (H. chrysoscelis and R. clamitans) or Harrison stage 2 (A. mexicanum) to time of hatching. Date of hatching and length at hatching were also compared between treatments for A. mexicanum. No significant differences in mortality between treatments were found between the three species. A. mexicanum in the 300 and 500 mg/L treatments hatched significantly earlier than individuals in the other treatment groups and, consequently, were significantly shorter in length at hatching. However, no effect on length was detected when days until hatching was considered as a covariate in the analysis. This study supports other recent research showing little to no effect of NO(3)- on amphibian embryos. The lack of effect at such high nitrate concentrations raises questions about the specific mechanisms responsible for protecting amphibians from NO(3)- during embryonic development, especially when compared to other chemicals that have shown more deleterious effects.