The sublethal effects of 2,4-D dimethylamine on wood frog tadpoles in Saskatchewan

Declining amphibian populations in association with an incidence of deformities have been observed globally. These observations have alarmed the scientific community as well as the general public. Potential causes include exposure to pesticides; therefore two experiments were performed to test the sublethal effects of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) on tadpoles of the wood frog (Rana sylvatica). Wood frog tadpoles in the first experiment were exposed to 2,4-D amine at 0.1, 1.0 and 100 ug/L in outdoor microcosms. Morphometric measures (total length, snout-vent length (SVL) and wet weight) were taken at metamorphic climax. Deformities and circulating hormone concentrations (corticosterone - CORT) were also assessed. Results showed that though tadpoles were exposed to various concentrations of 2,4-D throughout their aquatic life, there were no treatment differences associated with any of the endpoints except for total length (p = 0.023). Total length during metamorphosis was highly variable. Although statistically significant, biological significance was questionable. The second experiment was conducted in the field using natural ponds. The experimental groups included forested ponds (removed from pesticide exposure), agricultural ponds (potentially exposed) and treated agricultural ponds (intentionally treated with 2,4-D to achieve a concentration of 10 ug/L). Relatively rapid degradation of 2,4-D occurred in all treated ponds. The mean half-life was 8.0 + 5.5 days. In spite of the degradation, 2,4-D was present in the ponds until tadpoles metamorphosed. Similar endpoints including morphometric measures, as well as deformities and plasma CORT hormone were determined. In addition, total lipid and total protein (of the carcass) were also measured. Unlike the microcosm study, statistically significant differences were observed in SVL, wet weight and total protein although the differences may have been unrelated to 2,4-D exposure. The metamorphs sampled from the forested ponds were smaller in SVL (23% shorter) and wet weight (58% lighter) (p < 0.029) relative to the other two groups. Total protein in the metamorphs from the forested ponds was 22% lower than that of the agricultural ponds (p = 0.020). Reduced hormonal response to acute stress (p = 0.001) was found in metamorphs of the forested ponds compared to those of the other two groups (66% lower response). Herbicide exposure may have induced low level stimulation of growth (a positive impact on the exposed animals), as well as a possible elevation of baseline corticosterone, in the agricultural and treated metamorphs or there may have been a negative stressor present in the forested pond environment. It is not possible to identify the specific factors that were involved. It is unclear if 2,4-D affected the growth and development of wood frog tadpoles