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Water: Whole Effluent Toxicity

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

This manual describes six short-term (one-hour to nine-day) methods for estimating the chronic toxicity of effluents and receiving waters to five species:

  1. the sheepshead minnow, Cyprinodon variegatus
  2. the inland silverside, Menidia beryllina
  3. the mysid shrimp, Mysidopsis bahia
  4. the sea urchin, Arbacia punctulata
  5. and the red macroalga, Champia parvula

These rules do not apply to discharges into marine waters of the Pacific Ocean. 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) (486 pp, 7.0 MB)
  • Sections 1-3 (PDF) (30 pp, 338K)
    • Table of Contents
    • Introduction
    • Types of Tests
    • Health and Safety
  • Sections 4-10 (PDF) (46 pp, 777K)
    • Quality Assurance
    • Facilities, Equipment and Supplies
    • Test Organisms
    • Dilution Water
    • Sampling and Sample Handling
    • Data Analysis
    • Report Preparation
  • Section 11 (PDF) (62 pp, 1MB)
    • Test Method 1004.0 - Sheepshead minnow, Cyprinodon variegatus, Larval Survival and Growth
  • Section 12 (PDF) (638 pp, 90K)
    • Test Method 1005.0 - Sheepshead minnow, Cyprinodon variegatus, Embryo-larval Survival and Teratogenicity
  • Section 13 (PDF) (59 pp, 1.1MB)
    • Test Method 1006.0 - Inland Silverside, Menidia beryllina, Larval Survival and Growth
  • Section 14a (PDF) (27 pp, 882K) and Section 14b (PDF) (52 pp, 1.6MB)
    • Test Method 1007.0 - Mysid, Mysidopsis bahia, Survival, Growth, and Fecundity
  • Section 15 (PDF) (39 pp, 619K)
    • Test Method 1008.0 - Sea Urchin, Arbacia punctulata, Fertlization
  • Section 16 (PDF) (51 pp, 775K)
    • Test Method 1009.0 - Red Macroalga, Champia parvula, Reproduction
  • Appendices A-F (PDF) (49 pp, 355K)
    • 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-L (PDF) (33 pp, 552K)
    • 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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