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Water: Whole Effluent Toxicity

Short-Term Methods For Estimating the Chronic Toxicity of Effluents and Receiving Water to Freshwater Organisms

This manual describes four- to seven-day methods for estimating the chronic toxicity of effluents and receiving waters to three species:

  1. the fathead minnow, Pimephales promelas
  2. the cladoceran, Ceriodaphnia dubia
  3. the alga, Selenastrum capriconutum

Guidelines are included on laboratory safety, quality assurance, facilities and equipment, dilution water, effluent sampling methods and holding times and temperatures, data analysis, report preparation, and organism culturing and handling.

Browse Introduction

You will need Adobe Reader to view some of the files on this page. See EPA's PDF page to learn more.

  • Complete Document (PDF) (350 pp., 4.6 MB)
  • Table of Contents (PDF) (95 K, 12 pp)
  • Sections 1-6 (PDF) (557 K, 40 pp)
    • Table of Contents
    • Introduction
    • Short-Term Methods for Estimating Chronic Toxicity
    • Health and Safety
    • Quality Assurance
    • Facilities, Equipment and Supplies
    • and Test Organisms
  • Sections 7-10 (PDF) (520 K, 27 pp)
    • Dilution Water; Sampling, Sample Handling, and Preparation for Toxicity Tests
    • Data Analysis
    • Report Preparation
  • Section 11 (PDF) (1.2 MB, 59 pp)
    • Test method 1000.0 - Fathead minnow, Pimephales promelas
    • Embryo-larval Survival and Growth
  • Section 12 (PDF) (561 K, 29 pp)
    • Test method 1001.0 - Fathead minnow, Pimephales promelas
    • Embryo-larval Survival and Teratogenicity
  • Section 13 (PDF) (1.1 MB, 56 pp)
    • Test method 1002.0 - Daphnid, Ceriodaphnia dubia, Survival and Reproduction
  • Section 14 (PDF) (688 K, 47 pp)
    • Test method 1003.0 - Green alga, Selenastrum capriconutum, Growth; Bibliography
  • Appendices A-F (PDF) (283 K, 53 pp)
    • Independence, Randomization, and Outliers
    • Validating Normality and Homogeneity of Variance Assumptions
    • Dunnett's Procedure
    • T test with Bonferroni's Adjustment
    • Steel's Many-one Rank Test
    • Wilcoxon Rank Sum Test
  • Appendices G-M (PDF) (596 K, 39 pp)
    • Fisher's Exact Test
    • Single Concentration Toxicity Test
    • Probit Analysis
    • Spearman-Karber Method
    • Trimmed Spearman-Karber Method; Graphical Method
    • Linear Interpolation Method

Supporting Rule

Guidelines Establishing Test Procedures for the Analysis of Pollutants; Whole Effluent Toxicity Test Methods; Final Rule (November 19, 2002) In this final regulation, we ratified approval of several test procedures for measuring the toxicity of effluents and receiving waters. The test procedures are commonly referred to as whole effluent toxicity or WET test methods. We also withdrew two WET test methods from the list of nationally-approved biological test procedures for the analysis of pollutants. This action also revises some of the WET test methods to improve performance and increase confidence in the reliability of the results. This action satisfies settlement agreement obligations designed to resolve litigation over an earlier rulemaking that originally approved WET test methods.


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