Abstract. Methods recommended by Environment Canada for performing chronic three- brood toxicity tests with the freshwater cladoceran, Ceriodaphnia dubia, ...
EPS 1/RM/21 Second EditionBB February 2007 Environmental Science and Technology Centre Science and Technology Branch Environment Canada
ENVIRONMENTAL PROTECTION SERIES
Biological Test Method: B Test of Reproduction and Survival Using the Cladoceran Ceriodaphnia dubia
Environment Canada
Environnement Canada
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Agriculture Anaerobic Technology Airborne Pollutants Aquatic Toxicity Commercial Chemicals Consumers and the Environment Chemical Industries Federal Activities Food Processing Hazardous Wastes Inorganic Chemicals Marine Pollutants Mining and Ore Processing Northern and Rural Regions Paper and Fibres Power Generation Petroleum and Natural Gas Refrigeration and Air Conditioning Reference Methods Surface Finishing Oil and Chemical Spills Standard Reference Methods Transportation Systems Textiles Urban Pollution Wood Protection/Preservation
New subject areas and codes are introduced as they become necessary. A list of EPS reports may be obtained from Communications Services, Environment Canada, Ottawa, Ontario, K1A 0H3, Canada.
Biological Test Method: Test of Reproduction and Survival Using the Cladoceran Ceriodaphnia dubia
Method Development and Applications Section Environmental Science and Technology Centre Science and Technology Branch Environment Canada Ottawa, Ontario
Report EPS 1/RM/21 SECOND EDITION February 2007
Library and Archives Canada Cataloguing in Publication Biological test method. Test of reproduction and survival using the cladoceran Ceriodaphnia dubia/ Method Development and Applications Section, Environmental Science and Technology Centre, Environment Canada.–2nd ed.
(Report ; EPS 1/RM21) Includes abstract in French. Issued also in French under title: Méthode d’essai biologique. Essai de reproduction et de survie du cladocère Ceriodaphnia dubia. Includes bibliographical references: p. ISBN 0-662-42900-1 Cat. no.: En49-7/1-21E 1. Ceriodaphnia dubia--Effect of water pollution on. 2. Effluent quality--Testing--Standards--Canada. 3. Toxicity testing--Methodology--Standards--Canada. 4. Water quality bioassay. 5. Environmental toxicology. I. Canada. Environment Canada II. Environmental Science and Technology Centre (Canada). Method Development and Applications Section III. Series: Report (Canada. Environment Canada) ; EPS 1/RM/21
QL444.B83B56 2007
628.1'61
C2006-980124-X
© Her Majesty in Right of Canada (Environment Canada) 2006 ISBN 0-662-42900-1 Cat. no.: En49-7/1-21E
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Readers’ Comments Comments regarding the content of this report should be addressed to: Richard Scroggins, Chief Biological Methods Division Environmental Science and Technology Centre Environment Canada 335 River Road Ottawa, ON K1A OH3
Lisa Taylor, Manager Method Development & Applications Section Environmental Science and Technology Centre Environment Canada 335 River Road Ottawa, ON K1A 0H3
Cette publication est aussi disponible en français. Pour l’obtenir, s’adresser à: Services des communications Environnement Canada Ottawa (Ontario) K1A 0H3
Review Notice This report has been reviewed by the staff of the Environmental Technology Advancement Directorate, Environment Canada, and approved for publication. Mention of trade names or commercial products does not constitute endorsement by Environment Canada for use. Other products of similar value are available.
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Abstract Methods recommended by Environment Canada for performing chronic threebrood toxicity tests with the freshwater cladoceran, Ceriodaphnia dubia, are described in this report. This second edition of EPS 1/RM/21, published in 2007, supersedes the first edition that was published in 1992. It includes numerous procedural modifications as well as updated guidance and instructions to assist in performing the biological test method. General or universal conditions and procedures are outlined for undertaking this chronic toxicity test using a variety of test materials or substances. Additional conditions and procedures are stipulated which are specific for assessing samples of chemicals, effluents, elutriates, leachates, or receiving waters. Included are instructions on culturing conditions and requirements, food preparation, sample handling and storage, test facility requirements, procedures for preparing test solutions and test initiation, specified test conditions, appropriate observations and measurements, endpoints, methods of calculation, and the use of reference toxicants.
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Résumé Le présent document expose les méthodes recommandées par Environnement Canada pour l’exécution d’essais de toxicité chronique sur trois couvées du cladocère d’eau douce Ceriodaphnia dubia. Cette deuxième édition du document SPE 1/RM/21, publiée en 2007, remplace la première édition, parue en 1992. Elle comporte de nombreuses modifications procédurales, de même que des indications et des instructions à jour qui faciliteront l’exécution de la méthode d’essai biologique. Les conditions et procédures générales ou universelles décrites ici permettent de réaliser des essais de toxicité chronique avec un large éventail de matières ou de substances d’essai. Le document précise d’autres conditions et procédures propres à l’évaluation d’échantillons de substances chimiques, d’effluents, d’élutriats, de lixiviats ou d’eaux réceptrices. Il renferme aussi des instructions sur les conditions et règles d’élevage des organismes d’essai, la préparation des aliments, la manipulation et l’entreposage des échantillons, les exigences en matière d’installations d’essai, les procédures entourant la préparation des solutions d’essai et la mise en route des essais, les conditions prescrites pour les essais, les observations et les mesures pertinentes, les paramètres, les méthodes de calcul et l’utilisation de toxiques de référence.
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Foreword This is one of a series of recommended methods for measuring and assessing the toxic effect(s) on single species of aquatic or terrestrial organisms, caused by their exposure to samples of toxic or potentially toxic substances or materials under controlled and defined laboratory conditions. Recommended methods are those that have been evaluated by Environment Canada (EC), and are favoured: •
for use in EC environmental toxicity laboratories;
•
for testing that is contracted out by Environment Canada or requested from outside agencies or industry;
•
in the absence of more specific instructions, such as are contained in regulations; and
•
as a foundation for the provision of very explicit instructions as might be required in a regulatory protocol or standard reference method.
The different types of tests included in this series were selected because of their acceptability for the needs of programs for environmental protection and management carried out by Environment Canada. These reports are intended to provide guidance and to facilitate the use of consistent, appropriate, and comprehensive procedures for obtaining data on the toxicity to aquatic or terrestrial life of samples of specific test substances or materials destined for or within the environment. Depending on the biological test method(s) chosen and the environmental compartment of concern, substances or materials to be tested for toxicity could include samples of chemical or chemical product, effluent, elutriate, leachate, receiving water, sediment or similar particulate material, or soil or similar particulate material. Appendix G provides a listing of the biological test methods and supporting guidance documents published to date by Environment Canada as part of this series. Words defined in the Terminology section of this document are italicized when first used in the body of the report according to the definition. Italics are also used as emphasis for these and other words, throughout the report.
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Table of Contents Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v Résumé . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vi Foreword . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii List of Tables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xii List of Figures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xii List of Abbreviations and Chemical Formulae . . . . . . . . . . . . . . . . . . . . . . . xiii Terminology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xiv Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xxiii Section I Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1.1 Background . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1.2 Species Description and Historical Use in Tests . . . . . . . . . . . . . . . . . . . . 1 Section 2 Test Organisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 2.1 Species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 2.2 Life Stage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 2.3 Source . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 2.4 Culturing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 2.4.1 General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 2.4.2 Facilities and Apparatus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 2.4.3 Lighting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 2.4.4 Culture Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 2.4.5 Temperature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 2.4.6 Dissolved Oxygen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 2.4.7 pH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 2.4.8 Hardness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 2.4.9 Feeding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 2.4.10 Handling Organisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 2.4.11 Health Criteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Section 3 Test System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 3.1 Facilities and Apparatus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 3.2 Lighting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 3.3 Test Vessels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 3.4 Control/Dilution Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Section 4 Universal Test Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16 4.1 Preparing Test Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16 4.2 Beginning the Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
x 4.3 4.4 4.4.1 4.4.2 4.5 4.6 4.6.1 4.6.2 4.7 4.8
Renewing Test Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22 Test Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22 Dissolved Oxygen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23 pH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24 Test Observations and Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . 24 Test Endpoints and Calculations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 Multi-Concentration Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 Single-Concentration Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 Reference Toxicant . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 Legal Considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
Section 5 Specific Procedures for Testing Chemicals . . . . . . . . . . . . . . . . . . . . . . . . . . . 31 5.1 Properties, Labelling, and Storage of Sample . . . . . . . . . . . . . . . . . . . . . . 31 5.2 Preparing Test Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31 5.3 Control/Dilution Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 5.4 Test Observations and Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . 33 5.5 Test Endpoints and Calculations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 Section 6 Specific Procedures for Testing Effluent, Elutriate, and Leachate Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 6.1 Sample Collection, Labelling, Transport, and Storage . . . . . . . . . . . . . . . 35 6.2 Preparing Test Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 6.3 Control/Dilution Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 6.4 Test Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 6.5 Test Observations and Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 6.6 Test Endpoints and Calculations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 Section 7 Specific Procedures for Testing Receiving-Water Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 7.1 Sample Collection, Labelling, Transport, and Storage . . . . . . . . . . . . . . . 40 7.2 Preparing Test Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 7.3 Control/Dilution Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 7.4 Test Observations and Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 7.5 Test Endpoints and Calculations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 Section 8 Reporting Requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 8.1 Minimum Requirements for Test-Specific Report . . . . . . . . . . . . . . . . . . 42 8.1.1 Test Substance or Material . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 8.1.2 Test Organisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 8.1.3 Test Facilities and Apparatus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 8.1.4 Control/Dilution Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
xi 8.1.5 8.1.6 8.1.7 8.2 8.2.1 8.2.2 8.2.3 8.2.4 8.2.5 8.2.6 8.2.7
Test Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 Test Conditions and Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 Test Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 Additional Reporting Requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 Test Substance or Material . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 Test Organisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 Test Facilities and Apparatus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 Control/Dilution Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 Test Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 Test Conditions and Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Test Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 Appendix A Members of the Inter-Governmental Environmental Toxicity Group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53 Appendix B Environment Canada Regional and Headquarters’ Office Addresses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55 Appendix C Procedural Variations for Chronic Toxicity Tests with Ceriodaphnia spp., as Described in Canadian and U.S. Methodology Documents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Appendix D Procedure for Preparing YCT and Algal Food for C. dubia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 Appendix E Logarithmic Series of Concentrations Suitable for Use in Toxicity Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 Appendix F Analysis of Mortality to Estimate the Median Lethal Concentration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69 Appendix G Biological Test Methods and Supporting Guidance Documents Published by Environment Canada’s Method Development and Applications Section . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
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List of Tables 1
Checklist of Recommended Conditions and Procedures for Culturing Ceriodaphnia dubia . . . . . . . . . . . . . . . . . . . . . . 7
2
Preparation of Reconstituted Water of a Desired Hardness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
3
Checklist of Recommended Test Conditions and Procedures for Three-Brood Chronic Toxicity Tests with Ceriodaphnia dubia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
List of Figures 1
Diagram of Approach Taken in Delineating Test Conditions and Procedures Appropriate for Various Types of Test Materials or Substances . . . . . . . . . . . . . . . . . . 2
2
Anatomy of Female Ceriodaphnia dubia . . . . . . . . . . . . . . . . . . . . . . . . . . 6
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List of Abbreviations and Chemical Formulae ANOVA °C CaCO3 CaCl2 CaSO4 cm CoCl2 CuCl2 d DO EDTA FeCl3 g g/kg h H3BO3 HCl H2O ICp KCl K2HPO4 L LC LC50 LT50 LOEC m mg mS/m MgCl2 MgSO4 MnCl2
analysis of variance degree(s) Celsius calcium carbonate calcium chloride calcium sulphate centimetre(s) cobalt chloride copper chloride day(s) dissolved oxygen (concentration) ethylenediamine tetraacetate (C10H14O8N2) ferric chloride gram(s) gram(s) per kilogram hour(s) boric acid hydrochloric acid water inhibiting concentration for a (specific) percent effect potassium chloride potassium phosphate litre(s) lethal concentration median lethal concentration time to 50% mortality (lethality) lowest-observed-effect concentration metre(s) milligram(s) millisiemens per meter magnesium chloride magnesium sulphate manganous chloride
min mL mm mS N Na2EDTA NaHCO3 NaOH Na2MoO4 NaNO3 NOEC SD sp TM (TM) µg µE µmhos µm YCT ZnCl2 ZnSO4 >
