The fish toxicity assay most commonly used to establish chronic effects is the Organisation for Economic Co-operation and Development (OECD) 210, fish early-life stage test. However, the authors are not aware of any systematic analysis of the experimental design or statistical characteristics of the test since the test guideline was adopted nearly 20 years ago. Here, the authors report the results of an analysis of data compiled from OECD 210 tests conducted by industry labs. The distribution of responses observed in control treatments was analyzed, with the goal of understanding the implication of this variability on the sensitivity of the OECD 210 test guideline and providing recommendations on revised experimental design requirements of the test. Studies were confined to fathead minnows, rainbow trout, and zebrafish. Dichotomous endpoints (hatching success and posthatch survival) were examined for indications of overdispersion to evaluate whether significant chamber-to-chamber variability was present. Dichotomous and continuous (length, wet wt, dry wt) measurement endpoints were analyzed to determine minimum sample size requirements to detect differences from control responses with specified power. Results of the analysis indicated that sensitivity of the test could be improved by maximizing the number of replicate chambers per treatment concentration, increasing the acceptable level of control hatching success and larval survival compared to current levels, using wet weight measurements rather than dry weight, and focusing test efforts on species that demonstrate less variability in outcome measures. From these analyses, the authors provide evidence of the impact of expected levels of variability on the sensitivity of traditional OECD 210 studies and the implications for defining a target for future animal alternative methods for chronic toxicity testing in fish. Environ. Toxicol. Chem. 2012;31:370–376. © 2011 SETAC
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