Evaluating a developmental endocrine toxicity assay for Blanchard's cricket frog (Acris blanchardi) in outdoor enclosures.

A developmental toxicity testing design was evaluated for larval and post-metamorphic Blanchard's cricket frogs (Acris blanchardi) raised in outdoor enclosures. Larvae were chronically exposed to 17β-estradiol (0.0-2.3 μg/L E2) from free swimming (Gosner stage 26) until metamorphosis. Juvenile frogs were allowed to mature within the enclosures for 60 days to assess effects of larval exposure on development, including body mass, snout-vent length (SVL), sex ratio, gonad size, and gonadal histopathology. Forty-eight percent of the initial 600 animals were recovered at the end of the study. Recovery was not influenced by E2 exposure, but larval losses were negatively impacted by unusually high spring rain events that flooded some larval tanks, and heat-related mortality of late stage larvae during summer. All surviving larvae completed metamorphosis within an average of 47 days. Overall, E2 exposure did not influence sex ratio, or the body mass, SVL, or gonad size of either males or females. Development of testes was not influenced by E2 exposure, but oviduct development in males was 4.5-fold greater in the highest treatment. Oviduct and ovary development in females exposed to the two highest E2 treatments were half that of control females. Although not treatment related and despite ad-lib feeding, variation in terminal body mass and SVL within enclosures was pronounced, with minimum - maximum differences ranging from 207 to 1442 mg for body mass and 1 mm to 15 mm for SVL. This design allowed us to assess the effects of larval exposure to a contaminant on post-metamorphic development of a native amphibian in a semirealistic field environment. With modifications to decrease flooding or overheating, this enclosure design and species is a good test system for assessing contaminant effects on development of an amphibian from early larval stages through reproductive maturity.