Biological test method: fertilization assay using echinoids (sea urchins and sand dollars), chapter 1

Table of Contents

• Front Matter
  • Readers' Comments
  • Review Notice
  • Abstract
  • Foreword
  • List of Abbreviations and Chemical Formulae
  • Terminology
  • Acknowledgements
• Section 1: Introduction
  • 1.1 Background
  • 1.2 General Aspects of Echinoids and Their Use in Tests
• Section 2: Test Organisms
  • 2.1 Species
  • 2.2 Life Stage, Size, and Source
  • 2.3 Holding and Acclimating Adults in the Laboratory
    • 2.3.1 General
    • 2.3.2 Holding Containers
    • 2.3.3 Lighting
    • 2.3.4 Water
    • 2.3.5 Temperature
    • 2.3.6 Dissolved Oxygen
    • 2.3.7 pH
    • 2.3.8 Feeding
    • 2.3.9 Cleaning the Holding Containers
    • 2.3.10 Disease and Mortality
• Section 3: Test System
  • 3.1 Facilities and Apparatus
  • 3.2 Lighting
  • 3.3 Test Vessels
  • 3.4 Control/Dilution Water
• Section 4: Universal Test Procedures
  • 4.1 Preparing Test Solutions
    • 4.1.1 Control/Dilution Water
    • 4.1.2 Concentrations
    • 4.1.3 Replication
    • 4.1.4 Controls
  • 4.2 Beginning and Performing the Exposure
    • 4.2.1 Collecting Gametes for the Test
    • 4.2.2 Preparing Standard Suspensions of Gametes
    • 4.2.3 Ratio of Sperm to Eggs
    • 4.2.4 Exposure of Gametes
  • 4.3 Test Conditions
    • 4.3.1 Temperature
    • 4.3.2 Salinity
    • 4.3.3 Dissolved Oxygen and Aeration
    • 4.3.4 pH
  • 4.4 Test Observations and Measurements
  • 4.5 Test Endpoints and Calculations
    • 4.5.1 Validity of Test
    • 4.5.2 Multi-Concentration Tests
    • 4.5.3 Single Concentration Tests
  • 4.6 Reference Toxicant
  • 4.7 Legal Considerations
• Section 5: Specific Procedures for Testing Chemicals
  • 5.1 Properties, Labelling, and Storage of Sample
  • 5.2 Preparing Test Solutions
  • 5.3 Control/Dilution Water
  • 5.4 Test Observations and Measurements
  • 5.5 Test Endpoints and Calculations
• Section 6: Specific Procedures for Testing Samples of Effluent, Leachate, and Elutriate
  • 6.1 Sample Collection, Labelling, Transport, and Storage
  • 6.2 Preparing Test Solutions
  • 6.3 Control/Dilution Water
  • 6.4 Test Observations and Measurements
  • 6.5 Test Endpoints and Calculations
• Section 7: Specific Procedures for Testing Receiving-Water Samples
  • 7.1 Sample Collection, Labelling, Transport, and Storage
  • 7.2 Preparing Test Solutions
  • 7.3 Control/Dilution Water
  • 7.4 Test Observations and Measurements
  • 7.5 Test Endpoints and Calculations
• Section 8: Specific Procedures for Testing Samples of Liquid Extracted from Sediment or Similar Material
  • 8.1 General Aspects of Procedure
    • 8.1.1 Sample Labelling, Transport and Storage
    • 8.1.2 Preparing Samples
    • 8.1.3 Observations and Measurements on Sample
    • 8.1.4 Liquid from Control and Reference Sediments
  • 8.2 Testing Liquids Extracted from Sediments and Similar Solids
    • 8.2.1 Preparing Test Samples
    • 8.2.2 Control/Dilution Water
    • 8.2.3 Endpoints and Calculations
• Section 9: Reporting Requirements
  • 9.1 Minimum Requirements for Test-Specific Report
    • 9.1.1 Test Substance or Material
    • 9.1.2 Test Organisms
    • 9.1.3 Test Facilities and Apparatus
    • 9.1.4 Control/Dilution Water
    • 9.1.5 Test Method
    • 9.1.6 Test Conditions and Procedures
    • 9.1.7 Test Results
  • 9.2 Additional Reporting Requirements
    • 9.2.1 Test Substance or Material
    • 9.2.2 Test Organisms
    • 9.2.3 Test Facilities and Apparatus
    • 9.2.4 Control/Dilution Water
    • 9.2.5 Test Method
    • 9.2.6 Test Conditions and Procedures
    • 9.2.7 Test Results
• References
• Appendices
  1. Biological Test Methods and Supporting Guidance Documents Published by Environment Canada’s Method Development & Applications Unit
  2. Members of the Inter-Governmental Environmental Toxicity Group (as of October 2009)
  3. Environment Canada Regional and Headquarters’ Office Addresses
  4. Review of Procedural Variations Used by Previous Authors and Groups for Fertilization Assays Using Sea Urchins and Sand Dollars
  5. Bibliography. Additional Papers Directly Relevant to Canadian Echinoid Fertilization Assay
  6. Logarithmic Series of Concentrations Suitable for Toxicity Tests

List of Tables

1. General Features and Conditions for Spawning Echinoids to be Used as a Source of Gametes
2. Checklist of Recommended Conditions and Procedures for Holding and Acclimating Echinoid Adults
3. Checklist of Recommended and Required Test Conditions and Procedures
4. Summary of Sperm and Egg Additions to Each Test Vessel for the Three Test Volumes

List of Figures

1. Diagram of Approach Taken in Delineating Test Conditions and Procedures Appropriate for Various Types of Materials or Substances
2. General Appearance of Echinoids
3. Discriminating Between Fertilized and Unfertilized Eggs
4. Flowchart summarizing steps in statistical analysis of a multi-concentration test to derive an ICp