There are continued concerns about endocrine-disrupting chemical effects, and appropriate vertebrate models for assessment of risk are a high priority. Frog tadpoles are very sensitive to environmental substances because of their habitat and the complex processes of metamorphosis regulated by the endocrine system, mainly thyroid hormones. During metamorphosis, marked alteration in hormonal factors occurs, as well as dramatic structural and functional changes in larval tissues. There are a variety of mechanisms determining thyroid hormone balance or disruption directly or indirectly. Direct-acting agents can cause changes in thyroxine synthesis and/or secretion in thyroid through effects on peroxidases, thyroidal iodide uptake, deiodinase, and proteolysis. At the same time, indirect action may result from biochemical processes such as sulfation, deiodination and glucuronidation. Because their potential to disrupt thyroid hormones has been identified as an important consideration for the regulation of chemicals, the OECD and the EPA have each established guidelines that make use of larval African clawed frogs (Xenopus laevis) and frog metamorphosis for screening and testing of potential endocrine disrupters. The guidelines are based on evaluation of alteration in the hypothalamic-pituitary-thyroid axis. One of the primary endpoints is thyroid gland histopathology. Others are mortality, developmental stage, hind limb length, snout-vent length and wet body weight. Regarding histopathological features, the guidelines include core criteria and additional qualitative parameters along with grading. Taking into account the difficulties in evaluating amphibian thyroid glands, which change continuously throughout metamorphosis, histopathological examination has been shown to be a very sensitive approach.
The toxic effects of pesticides on earthworms, one of the most important bioindicators in the terrestrial environment, are closely related to their body burden determined by uptake, metabolism and excretion processes. Not only the passive diffusion via the outer skin from a dissolved fraction of pesticide but also the ingestion of contaminated soil and food governs the uptake process, with each contribution controlled by either the hydrophobicity of the pesticide or the soil organic matter. Although the available information is limited, earthworms are likely to metabolize pesticides via hydrolysis and oxidation (Phase I) followed by conjugation (Phase II), and low bioaccumulation is observed as a result for most pesticides. The acute toxicity in the soil exposure can be partly explained by the dissolved fraction of pesticide in pore water, but the contribution of dietary uptake and metabolism should be further studied to correctly evaluate pesticide toxicity.
Exposure to pesticide residues is claimed to be one of the possible causes of frog decline. Knowledge of basic information on the uptake, metabolism and depuration processes of pesticides in the frog is needed to understand the relationship between exposure and toxic effects from their actual body burden, together with their bioconcentration. The hydrophobicity of pesticides and industrial chemicals was one of the most important factors controlling bioconcentration, similarly to fish, when frogs are exposed to contaminated water. Skin absorption was also a key route in the uptake process especially in the adult frog. The metabolic profiles in the frog, mainly examined by an intraperitoneal injection technique, were common to other aquatic species without any frog-specific transformation reaction. The effects of developmental stage, sex, species and environmental factors such as temperature were observed for bioconcentration and metabolism.
Bioconcentration and metabolism of pyriproxyfen uniformly labeled with C at the phenoxyphenyl ring were studied using tadpoles of African clawed frog, Xenopus laevis, exposed to water at the nominal concentrations of 3 and 300 ppb for 22 days under the flow-through conditions, with a following 3 day depuration phase. Neither meaningful mortality nor abnormal behavior was observed in control and exposure groups throughout the study. After the rapid uptake to tadpoles, pyriproxyfen was extensively metabolized and excreted, and as a result, steady-state bioconcentration factors and depuration half-lives ranged from 550 to 610 and from 0.34 to 0.54 days, respectively. The metabolites were mostly distributed in the liver or gastrointestinal tract. The major metabolic reactions were hydroxylation at the 4' position of the phenoxyphenyl group and cleavage of the ether linkage, followed by sulfate conjugation.
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