Fathead minnow (Pimephales promelas) lifecycle tests have been used to assess the effects of many North American pulp mill effluents. Fish are exposed under controlled laboratory conditions to final effluent, from the egg stage through hatching, juvenile stage and mature adult stage. Outlined here are methods for the lifecycle test (which takes from 4 to 5 months to complete) with sampling of juvenile fish at 1, 2 and 3 months of age, and sampling of mature adults after breeding. The results of most fathead minnow lifecycle studies have shown that pulp mill effluents cause metabolic and reproductive disruption (enlarged livers, reduced egg production, smaller gonads, decreased sex hormones and reduced secondary sex characteristics). Thus, the fathead minnow lifecycle assay is able to mimic the most commonly observed changes seen in wild fish exposed to pulp mill effluents. Sensitive indicators of reproductive effects in fathead minnows exposed for a lifecycle to pulp mill effluents include secondary sex characteristics, time to first reproduction and number of eggs laid. Egg production is often the most sensitive response to pulp mill effluents. Because of the length and cost of full lifecycle tests, a shortened assay using adult fish was developed as a screen for endocrine-disrupting compounds. This assay, the terminal reproduction test, has been used with success on a few pulp mill effluents. The assay exposes fathead minnow breeding pairs for three weeks to effluent, and compares egg production, sex characteristics and bioindicators of reproductive performance in pre-exposure versus post-exposure fish. For assessment of pulp mill effluents, it appears that the sensitivity of the shortened terminal reproductive fathead minnow assay may be improved by assessing bioindicators of reproductive performance (such as circulating levels of sex hormones, and circulating or hepatic vitellogenin) along with changes in secondary sex characteristics and egg production.

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