EPA Method 1000

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EPA Method 1000:
Larval Survival and Growth Test using Fathead Minnow. Official Name: EPA Test Method 1000.1: Fathead Minnow, Pimephales promelas, Larval Survival and Growth Test Method

Fathead Minnow, Pimephales promelas, larvae are exposed in a static renewal system for seven days to different concentrations of effluent or to receiving water. Test results are based on the survival and weight of the larvae.

This method estimates the chronic toxicity of effluents and receiving water to the fathead minnow, Pimephales promelas, using newly hatched larvae in a seven-day, static renewal test. The effects include the synergistic, atagonistic, and additive effects of all the chemical, physical, and biological components which adversely affect the physiological and biochemical functions of the test organisms.

Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms

Introduced toxic substances; Low D.O.; High concentrations of suspended and/or dissolved solids; Pathogenic and/or predatory organisms;

QC Requirements:
Reference toxicant testing with control charts for each combination of toxicant, test species, test condition, and endpoint.

Maximum Holding Time:


Chronic Toxicity

Effluents: 5 and a control Receiving waters: 100% receiving water and a control (recommended)

Sample Prep:
Filtration, Dechlorination.

It is a laboratory's responsibility to demonstrate its ability to obtain consistent, precise results with reference toxicants before it performs toxicity tests with effluents for permit compliance purposes. To meet this requirement, the intra-laboratory precision, expressed as percent coefficient of variation, of each type of test to be used in a laboratory should be determined by performing five or more tests with different batches of test organisms using the same reference toxicant, at the same concentrations, with the same test conditions, and the same data analysis methods.


Revision Number:
Fourth Edition, October 2002