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Ecological Exposure Research

Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to West Coast Marine and Estuarine Organisms

This manual describes six short-term (forty minutes to seven days) estuarine and marine methods for measuring the chronic toxicity of effluents and receiving waters to eight species: the topsmelt, Atherinops affinis; the mysid, Holmesimysis costata; the sea urchin, Strongylocentrotus purpuratus and sand dollar Dendraster excentricus; the red abalone Haliotis rufescens; the bivalves Crassostrea gigas and mussel Mytilus spp. and the giant kelp, Macrocystis pyrifera. The methods include single and multiple concentration static renewal and static nonrenewal toxicity tests for effluents and receiving waters. Also included are guidelines on laboratory safety, quality assurance, facilities, and equipment and supplies; dilution water; effluent and receiving water sample collection, preservation, shipping, and holding; test conditions; toxicity test data analysis; report preparation; and organism culturing, holding, and handling. Examples of computer input and output for Dunnett's Procedure, Probit Analysis, Trimmed Speaman-Karber Method, and the Linear Interpolation Method are provided in the Appendices.

Document Introduction

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  • Contents (PDF, 15 pp., 29 KB)
    Foreward; Preface; Abstract; Acknowledgements; Table of Contents
  • Sections 1-3 (PDF, 13 pp., 27 KB)
    Introduction; Short-term Methods for Estimating Chronic Toxicity; Health and Safety
  • Sections 4-10 (PDF, 57 pp., 258 KB)
    Quality Assurance; Facilities, Equipment and Supplies; Test Organisms; Dilution Water; Sampling, Sample Handling, and Preparation for Toxicity Tests; Test Endpoints and Data Analysis; Report Preparation
  • Section 11 (PDF, 69 pp., 324KB)
    Topsmelt, Atherinops affinis; 7-Day Larval Growth and Survival Test Method
  • Section 12 (PDF, 69 pp., 329KB)
    Mysid, Holmesimysis costata; Survival and Growth Test Method
  • Section 13 (PDF, 50 pp., 177KB)
    Pacific Oyster, Crassostrea Gigas, and Mussel, Mytilus spp; Embryo-Larval Development Test
  • Section 14 (PDF, 59 pp., 191KB)
    Red Abalone, Haliotis rufescens; Larval Development Test Method
  • Section 15 (PDF, 68 pp., 2.38MB)
    Purple Urchin, Strongylocentrotus purpuratus and Sand Dollar, Dendraster excentricus; Larval Development Test Method
  • Section 16 (PDF, 77 pp., 2.81MB)
    Purple Urchin, Strongylocentrotus purpuratus and Sand Dollar, Dendraster excentricus; Fertilization Test method
  • Section 17 (PDF, 62 pp., 278KB)
    Giant Kelp, Macrocystis pyrifera; Germination and Germ-Tube Growth Test Method
  • References (PDF, 21 pp., 39KB)
  • Bibliography (PDF, 15 pp., 36KB)
  • Appendices (PDF, 98 pp., 217KB)
    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; Single Concentration Toxicity Test - Comparison of Control with 100% Effluent or Receiving Water; Probit Analysis; Spearman-Karber Method; Trimmed Spearman-Karber Method; Graphical Method; Linear Interpolation Method

Tropical Collector Urchin, Tripneustes gratilla,  Fertilization Test Method

This fertilization method estimates the chronic toxicity of effluents and receiving waters to the gametes of the tropical sea urchin (Tripneustes gratilla). The test uses a static, non-renewal 60-minute sperm exposure and a subsequent 20-minute fertilization period, following the addition of eggs, for measuring the fertilizing capacity of the sperm. The purpose of the test is to determine the concentrations of a test substance that reduce fertilization of exposed gametes, relative to that of the control.

Tropical Collector Urchin, Tripneustes gratilla, Fertilization Test Method,     EPA/600/R-12/022.  April 2012.

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