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The Japanese medaka (Oryzias latipes) early-life-stage bioassay (18-day test) provides a suitable laboratory model for the evaluation of toxicant impact. The naturally high variability in the time to hatch for medaka embryos is, however, a major limitation in terms of the duration of the test. In this study, the 18-day test was modified to use agitation to synchronize hatch and reduce time to hatch for nonexposed embryos. Then, bioassays were conducted using two different complex mixtures (polycyclic aromatic hydrocarbons (PAHs) and naphthenic acids (NAs)) to compare the sensitivity of the measurement endpoints using the 18-day standard protocol (SP) and the modified protocol (MP). Agitation reduced time to hatch by one third or more and improved hatch success (100%). The MP proved to be a more sensitive test method for the sublethal evaluation of a stressor (PAH mixture) that caused reduced hatch length and induced signs of blue sac disease (BSD) including heart deformities, yolk sac-pericardial edema, and cranial-skeletal deformities. In contrast, there was some variability in the sensitivity of the test methods for developmental endpoints for the sublethal evaluation of a stressor (NA mixture) that had no effect on hatch length and was a weak inducer of signs of BSD. Even though the sensitivity of the measurement endpoints were slightly more or less sensitive for the MP vs. SP depending on the mixture tested, the MP using agitation is recommended as a cost-effective and rapid alternative for screening the sublethal impact of toxicants on the early-life stages of fish.

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