Subacute effects of pulsed copper, zinc, or ammonia exposures were examined, including a range of pulse concentrations, durations, frequencies, and recovery times between pulses, using short-term chronic Pimephales promelas and 21-d Daphnia magna tests. Sublethal effects were rarely observed independent of mortality. Effects were observed only at concentrations near the species continuous exposure 48 h LC50 for each chemical. Daphnia often rebounded from temporary reproduction effects, meeting or exceeding control responses by the end of the test. Effects of 24 h ammonia or copper pulses were diminished soon after the pulse was removed, while 24 h zinc pulses caused continued effects for several days following removal of the pulse, indicating a slower uptake and/or depuration rate for zinc. D. magna exhibited less mortality as copper pulses were spaced further apart, while fish were equally or more affected with longer recovery times between copper pulses, indicative of different adaptation mechanisms between the two species. Responses were not predictable based on either average concentration or a combination of duration and concentration. Chronic water quality criteria and effluent permit limits, expressed as a 4- or 30-d average concentration, respectively, may not be appropriate for protecting against effects of pulsed exposures, depending on the frequency, magnitude, and duration of pulses, as well as the recovery period between events.
The U.S. Environmental Protection Agency (U.S. EPA) and state agencies implement the Clean Water Act, in part, by evaluating the toxicity of effluent and surface water samples. A common goal for both regulatory authorities and permittees is confidence in an individual test result (e.g., no-observed-effect concentration [NOEC], pass/fail, 25% effective concentration [EC25]), which is used to make regulatory decisions, such as reasonable potential determinations, permit compliance, and watershed assessments. This paper discusses an additional statistical approach (test of significant toxicity [TST]), based on bioequivalence hypothesis testing, or, more appropriately, test of noninferiority, which examines whether there is a nontoxic effect at a single concentration of concern compared with a control. Unlike the traditional hypothesis testing approach in whole effluent toxicity (WET) testing, TST is designed to incorporate explicitly both α and β error rates at levels of toxicity that are unacceptable and acceptable, given routine laboratory test performance for a given test method. Regulatory management decisions are used to identify unacceptable toxicity levels for acute and chronic tests, and the null hypothesis is constructed such that test power is associated with the ability to declare correctly a truly nontoxic sample as acceptable. This approach provides a positive incentive to generate high-quality WET data to make informed decisions regarding regulatory decisions. This paper illustrates how α and β error rates were established for specific test method designs and tests the TST approach using both simulation analyses and actual WET data. In general, those WET test endpoints having higher routine (e.g., 50th percentile) within-test control variation, on average, have higher method-specific α values (type I error rate), to maintain a desired type II error rate. This paper delineates the technical underpinnings of this approach and demonstrates the benefits to both regulatory authorities and permitted entities.
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