UC Berkeley

Agrochemical exposure is one of the factors that contributes to worldwide amphibian declines. Most studies that examine agrochemicals and amphibian declines focus on toxicity. However, declines are more likely caused by the sub-lethal effects of agrochemical exposure. Past emphases on the lethal effects of agrochemical exposure have overshadowed the contribution of decreased recruitment in amphibian declines. Additionally, studies that examine agrochemicals and reproductive function tend to focus on the effects of single chemical exposures instead of the effects of ecologically relevant mixtures. To address these issues, this dissertation examined the effects of ecologically relevant agrochemical exposures on the stress response and the reproductive endocrinology, morphology, and behaviors of male amphibians in the laboratory and the wild.

Chapter 1 provides a general review of the factors implicated in amphibian declines and provides an overview of the previous research conducted on the effects of agrochemical exposure on recruitment.

Chapter 2 is a field study that examined whether agricultural run-off alters the stress response and reproductive function of male bullfrogs (Lithobates catesbeianus). Bullfrogs were collected upstream and downstream of agricultural activity across three California river systems (Salinas, Sacramento and San Joaquin). Size, primary and secondary sex traits, sperm count, and corticosterone and testosterone levels were examined. Overall, bullfrogs living downstream of agricultural activity (i.e. exposure to agricultural run-off) were small and had elevated testosterone and corticosterone levels. In addition, downstream males from the Salinas and San Joaquin Rivers were also small in size and had elevated testosterone levels. However, only downstream males of the San Joaquin River had elevated corticosterone and exaggerated secondary sex traits. Together, these data suggest that living downstream of agriculture can alter size, hormone levels, and the expression of sexually dimorphic sex traits. Such changes to the reproductive endocrinology and morphology of male amphibians can be detrimental to the reproductive health and long-term reproductive success of amphibian populations.

In Chapter 3, I examined corticosterone, testosterone, and the reproductive clasping behaviors of adult male African clawed frogs (Xenopus laevis) exposed to field collected and simulated agricultural run-off. This experiment implemented a novel eco-relevant experimental design to mimic real-life agrochemical exposures. Male frogs were exposed to field water collected downstream (agricultural run-off) and upstream (negative control) of agricultural activity along the Salinas River, CA. In addition, a pesticide mixture containing the top agrochemicals used in the Monterey County was included to simulate agricultural run-off. Mating behavior was suppressed in males exposed to simulated agricultural run-off but enhanced in males exposed to field collected agricultural run-off. In addition, testosterone levels of clasping males were elevated in comparison to controls. Males immersed in simulated agricultural run-off had significantly lower testosterone levels than control males in 2010. These data suggest that agrochemical exposure (both field collected and simulated) can alter reproductive hormones and clasping behaviors. Altered sex hormones and behaviors in male amphibians may play a role in amphibian declines.

Lastly, this dissertation is summarized in Chapter 4. The applicability of this dissertation as a model for amphibian declines and other reproductive related human health concerns are also introduced.