EPA Method 1004

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EPA Method 1004:
Larval Survival and Growth Test using Sheepshead Minnow. Official Name: EPA Test Method 1004.0, Sheepshead Minnow, Cyprinodon variegatus, Larval Survival and Growth Test Method.

Sheepshead Minnow, Cyprinodon variegatus, larvae (preferably less than 24h old) 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, adapted in part from USEPA (1987b), estimates the chronic toxicity of effluents and receiving waters to the sheepshead minnow, Cyprinodon variegatus, using newly hatched larvae in a seven-day, static-renewal test. The effects include the synergistic, antagonistic, and additive effects of all the chemical, physical, and biological components which adversely affect the physiological and biochemical functions of the test species.

USEPA, October 2002, Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms. Third Edition. EPA-821-R-02-014.

Introduced toxic substances from testing equipment; Low D.O., high concentration of dissolved or suspended solids; Extremes of pH; Pathogenic or predatory organisms

QC Requirements:
Acceptable side-by-side reference toxicant testing; 80% or greater survival in controls, with a 0.50 mg average dry weight at conclusion of test.

Maximum Holding Time:
Sample must be used within 24h


Chronic Toxicity

Effluent: 5 and control Receiving water: 100% receiving water (or minimum of 5) and a control

Sample Prep:

Based on single and multi-laboratory precision data.


Revision Number:
Third Edition, October 2002

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Examples:  8260

Examples:  Dioxin
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