Endocrine Disrupters: Hazard Testing and Assessment Methods

Enables researchers to assess the effects of endocrinedisrupters as well as comply with new environmentalregulations

Endocrine disrupters are chemicals both man–made andnatural that interfere with the body′s endocrine system,potentially resulting in adverse developmental, reproductive,neurological, and immune effects. In recent years, a number ofregulatory authorities around the world have drafted or enactedlegislation that requires the detection and assessment of theeffects of endocrine disrupters on both humans and wildlife. Inresponse, this book provides comprehensive, up–to–date informationon the latest tested and proven methods used to detect and assessthe environmental hazards posed by endocrine–disruptingchemicals.

Endocrine Disrupters is divided into chapters coveringeach major taxon as well as chapters dedicated to hazard assessmentand regulation. The book covers testing methods for all thevertebrate groups and several invertebrate phyla, including:

Moreover, the book emphasizes practical, ethical testing methodsthat combine sensitivity, efficiency, statistical power, andreasonable cost.

Each chapter is written by one or more international experts inecotoxicology, offering readers step–by–step guidance forimplementing each method based on the latest research and theauthors′ firsthand laboratory experience. Furthermore, all thechapters have been subjected to a rigorous peer review and editedin light of the reviewers′ comments. References at the end of eachchapter guide readers to the literature in the field.

Endocrine Disrupters is recommended for scientists whoneed to test chemicals for possible endocrine–disruptingproperties. It is also recommended for regulatory authorities whoneed to decide whether particular chemicals can be safelymarketed.

Preface ix

Contributors xi

1 Ecotoxicity Test Methods for Endocrine–DisruptingChemicals: An Introduction 1
Peter Matthiessen

1.1 Background 1

1.2 Regulatory Concerns 2

1.3 Invertebrates 2

1.4 Vertebrates 3

1.5 Testing Schemes for EDCs 5

Reference 6

2 Endocrine Disruption inWildlife: Background, Effects, andImplications 7
Dick Vethaak and Juliette Legler

2.1 Background to Endocrine Disruption 8

2.2 Effects of EDCs on Wildlife 19

2.3 Weight of Evidence and Ecological Significance of ED Effects32

2.4 Implications for Effect Assessment and Toxicity Testing36

2.5 Need for More Field Studies and an Integrated Approach38

2.6 Concluding Points 39

References 40

3 The Regulatory Need for Tests to Detect EDCs and AssessTheir Hazards toWildlife 59
Hans–Christian Stolzenberg, Tobias Frische, Vicki L. Dellarco,Gary Timm, Anne Gourmelon, Taisen Iguchi, Flemming Ingerslev, andMike Roberts

3.1 Emerging Concerns and Policy Responses: Focusing on EDCs asa Large Pseudo–Uniform Group of Substances 60

3.2 General Approaches in Substance–Related RegulatoryFrameworks (EU) 80

3.3 How to Make EDC Definitions Operational forSubstance–Related Regulatory Work 87

3.4 Future Perspectives 91

3.5 Conclusions 92

References 93

4 Techniques for Measuring Endocrine Disruption in Insects100
Lennart Weltje

4.1 Introduction 100

4.2 Methods 105

4.3 Discussion 108

4.4 Conclusion 110

4.5 Acknowledgments 110

References 110

5 Crustaceans 116
Magnus Breitholtz

5.1 Introduction 116

5.2 Background to Crustacean Endocrinology 118

5.3 State of the Art: What Do We Know About Endocrine Disruptionin Crustaceans? 121

5.4 Available Subchronic/Chronic Standard Test Protocols 128

5.5 Complementary Tools for Identification of EndocrineDisruption 129

5.6 Summary and Conclusions 132

References 134

6 Endocrine Disruption in Molluscs: Processes and Testing143
Patricia D. McClellan–Green

6.1 Background and Introduction 143

6.2 What Constitutes the Endocrine System in Molluscs? 145

6.3 End Points and Biomarkers of Endocrine Disruption 154

6.4 Current Test Methods Using Molluscs 164

6.5 Proposed Test Methods 167

6.6 Conclusions 171

References 172

7 Using Fish to Detect Endocrine Disrupters and Assess TheirPotential Environmental Hazards 185
Peter Matthiessen

7.1 Introduction 185

7.2 International Efforts to Standardize Fish–Based Methods forScreening and Testing Endocrine–Disrupting

7.3 Fish–Based Screens Developed by OECD forEndocrine–Disrupting Chemicals 189

7.4 Progress with Developing Fish Partial Life Cycle Tests forEndocrine Disrupters 194

7.5 Prospects for the Standardization of Fish Full Life Cycleand Multigeneration Tests 195

7.6 Strengths and Weaknesses of a Hazard Evaluation StrategyBased Partly on Available and Proposed Fish Screens and Tests197

7.7 Conclusions 198

References 198

8 Screening and Testing for Endocrine–Disrupting Chemicals inAmphibian Models 202
Daniel B. Pickford

8.1 Introduction 202

8.2 Potential Uses of Amphibians in Endocrine DisrupterScreening and Testing Programs 203

8.3 Embryonic Development 205

8.4 Hatching 208

8.5 Larval Development 209

8.6 Higher–Tier Tests with Amphibians 224

8.7 Other and Emerging Test Methods 227

8.8 Summary and Conclusions 229

References 232

9 Endocrine Disruption and Reptiles: Using the UniqueAttributes of Temperature–Dependent Sex Determination to AssessImpacts 245
Satomi Kohno and Louis J. Guillette, Jr.

9.1 Introduction 245

9.2 Approaches to Examine Effects of EDCs 252

9.3 Induction of Sex Reversal In Ovo 255

9.4 Analysis of Sex–Reversed Animals 260

9.5 Conclusions 265

References 266

10 Birds 272
Paul D. Jones, Markus Hecker, Steve Wiseman, and John P.Giesy

10.1 Introduction 272

10.2 Differences Between Birds and Mammals and Among BirdSpecies 275

10.3 In Vitro Techniques 278

10.4 Studies with Embryos 280

10.5 In Vivo Techniques 280

10.6 Examples of EDC Effects from Field Studies 285

10.7 Proposed Two–Generation Test 288

10.8 Conclusions 291

References 292

11 Mammalian Methods for Detecting and AssessingEndocrine–Active Compounds 304
M. Sue Marty

11.1 Introduction 304

11.2 Mammalian Tier 1 Screening Assays 306

11.3 Tier 2 Tests 326

11.4 Human and Wildlife Relevance of Estrogen, Androgen, andThyroid Screening Assays 329

11.5 Potential Future Assays for Endocrine Screening 330

References 332

12 Application of the OECD Conceptual Framework for Assessingthe Human Health and Ecological Effects of Endocrine Disrupters341
Thomas H. Hutchinson, Jenny Odum, and Anne Gourmelon

12.1 Introduction 342

12.2 Overview of the OECD Revised CF 343

12.3 Application of the Klimisch Criteria to the EE2 and VINCase Studies 346

12.4 Case Study: Data Examples for 17–Ethynylestradiol 346

12.5 Case Study: Data Examples for Vinclozolin 357

12.6 Conclusions 367

References 368

13 The Prospects for Routine Testing of Chemicals forEndocrine–Disrupting Properties and Potential Ecological Impacts373
Peter Matthiessen

13.1 Introduction 373

13.2 Are There Gaps in the Test Suite for EDCs? 374

13.3 New Modes of Endocrine–Disrupting Action376

13.4 How Should Tests for EDCs Be Deployed in an IntegratedFashion? 377

13.5 Use of Weight of Evidence when Assessing Possible EDCs380

13.6 Conclusions 382

References 382

Index 385

PETER MATTHIESSEN, PhD, is an aquatic ecotoxicologist whoworks as an independent consultant. Specializing in the study ofendocrine disrupters, he has conducted extensive research into howsewage effluents cause feminization in wild male fish as well ashow tributyltin–based antifoulants cause masculinization in wildfemale mollusks. Professor Matthiessen is a member of the UKAdvisory Committee on Pesticides and Co–chair of the OECDValidation Management Group for Ecotoxicity Tests. He hascontributed to the development of standardized ecotoxicity assayswith sensitivity for endocrine disrupters as well as writtenguidance for the interpretation of the resulting data.