Ecotoxicology and Genotoxicology: Non-traditional Terrestrial Models (Issues in Toxicology, Volume 32)

The potential impact of anthropogenic pollutants such as agrochemicals on the environment is of global concern. Increasing use of certain compounds can result in contamination of food, water and atmospheric systems and in order to combat this pollution it is important to be able to accurately monitor the short and long term effects. This book describes the latest non-traditional terrestrial species models used as indicators of the toxic effects of environmental pollutants. The book enables understanding of the effects of pollutants in non-target species, and therefore enables analysis of the effects on ecosystems. This book will be of interest to anyone interested in developing new biomarker species with high degrees of ecological relevance. It will serve as a useful resource for regulatory and research toxicologists, particularly those interested in soil screening and the effects of pollutants on wildlife and insects and their use as biological indicators. Cover 1 Contents 12 Preface 8 Section I: Terrestrial Invertebrates as Experimental Models 20 Chapter 1 The Use of Non-standardized Invertebrates in Soil Ecotoxicology 22 1.1 Soil Invertebrates 22 1.2 The Use of Invertebrates in Soil Ecotoxicology 25 1.3 Key Groups of Invertebrates for Soil Ecotoxicological Testing 32 1.3.1 Earthworms 32 1.3.2 Collembolans 35 1.3.3 Enchytraeids 36 1.3.4 Isopods 37 1.3.5 Others 38 Acknowledgments 38 References 39 Chapter 2 Higher-tier Multi-species Studies in Soil: Prospects and Applications for the Environmental Risk Assessment of Pesticides 50 2.1 Introduction 50 2.2 Ecological Relevance of Soil Organisms in Agro-ecosystems 52 2.2.1 Structure and Function of Soils and Soil Organism Communities 52 2.2.2 Losses of Soil Biodiversity in Agricultural Landscapes 54 2.3 Status Quo and Developments of Risk Assessment for In-soil Organisms 54 2.3.1 Status Quo 54 2.3.2 Transition 55 2.3.3 New Developments 56 2.3.4 Challenges 57 2.3.5 Future Demands 58 2.4 Methodologies for Multispecies Tests in Soil 59 2.4.1 Ontology and History of Test Systems 59 2.4.2 Methodological Challenges of Multispecies Tests 62 2.5 Exposure of Soil Organisms Resulting From the Fate of Pesticides 65 2.6 Calibration of Soil Risk Assessment Using Semi-field Studies as Surrogate Reference Tiers 67 2.6.1 Specific Protection Goals 67 2.6.2 Derivation of Assessment Factors 68 2.6.3 TME as Surrogate Reference Tier 69 2.7 Conclusions 70 References 70 Chapter 3 Aporrectodea longa (Annelida, Lumbricidae): A Suitable Earthworm Model for Genotoxicity Evaluation in the Environment 78 3.1 Introduction 78 3.2 Experimental 81 3.2.1 Earthworm Collection and Storage 81 3.2.2 Soil Collection and Amendment 81 3.2.3 Exposure Following Amendment with Differing Pesticides or B[a]P 81 3.2.4 Coelomic Fluid Collection 82 3.2.5 The Alkaline Single Cell-gel Electrophoresis (‘comet') Assay 82 3.2.6 Exposure to Aged Cypermethrin Residues 83 3.3 Results and Discussion 83 3.3.1 Comet Generation from Differing Compounds 85 3.3.2 Changes in Comet Formation Following Exposure to Aged Cypermethrin Residues 87 3.3.3 Uptake of 14C-Compound 90 3.4 Conclusion 90 Acknowledgments 91 References 91 Chapter 4 Evaluation of the Genotoxic Potential of Contaminated Soil Employing the Snail Helix aspersa 95 4.1 Introduction 95 4.2 The Major Groups of Soil Contaminants 97 4.2.1 Heavy Metal 97 4.2.2 Organic Contaminants 98 4.2.3 Sewage Sludge 99 4.3 Helix aspersa for Biomonitoring of Contaminated Soil 99 4.4 Genotoxicity Tests with H. aspersa and Contribution to Environmental Research 103 4.5 Conclusions 107 Acknowledgments 108 References 108 Chapter 5 The Use of Spiders in the Assessment of Cellular Effects of Environmental Stressors 115 5.1 Introduction 115 5.2 Spiders in Ecosystems Contaminated with Heavy Metals 117 5.2.1 Cellular Defence Reactions in Spiders from Areas Affected by Industrial Pollution 120 5.3 Spider Sensitivity to Pesticides 126 5.3.1 Changes in AChE Activity 128 5.3.2 Enzymatic Detoxifying Reactions 129 5.3.3 Genotoxic and Cytotoxic Effects of Plant Protection Agents in Spiders 130 5.4 Starvation Stress 133 5.5 Conclusions 134 References 135 Section II: Terrestrial Vertebrates as Experimental Models 142 Chapter 6 Use of Melanin-pigmented Cells as a New Tool to Evaluate Effects of Agrochemicals and Other Emerging Contaminants in Brazilian Anurans 144 6.1 Color in Animals 144 6.2 Internal Melanin-pigmented Cells 147 6.3 Environmental Contamination and Its Effects on Visceral Pigmentation 148 6.4 Response of Cutaneous Melanocytes to Aquatic Contaminants 148 6.5 Response of Internal Melanocytes to Aquatic Contaminants 149 6.6 Response of Melanomacrophages to Aquatic Contaminants 153 6.7 Conclusion 157 Acknowledgments 157 References 157 Chapter 7 The Use of Terrestrial Life-stages of European Amphibians in Toxicological Studies 162 7.1 Introduction 162 7.2 Toxicological Studies on the Impact of Pesticides on Terrestrial Life-stages of European Amphibians 164 7.3 Risk Assessments for Terrestrial Life-stages of Amphibians in Pesticide Approval 168 7.3.1 Surrogate Species for Terrestrial Life-stages of Amphibians 169 7.3.2 Indirect Effects 174 7.4 Pesticide Formulations—Toxicity in the Mix? 174 7.5 Conclusions 175 Acknowledgments 176 References 176 Chapter 8 Impacts of Agriculture and Pesticides on Amphibian Terrestrial Life Stages: Potential Biomonitor/Bioindicator Species for the Pampa Region of Argentina 182 8.1 Introduction 182 8.2 Amphibian Diversity, Life History and Global Declines 183 8.2.1 Amphibian Diversity and Life History 183 8.2.2 Amphibian Declines 184 8.3 The Pampa Region of Argentina 185 8.3.1 Location, Geography and Characteristics 185 8.3.2 Evolution of Agricultural Practices and Environmental Impacts 186 8.4 Agriculture and Amphibian Declines: The Need for Biomonitoring 188 8.4.1 Agriculture and Amphibian Declines 188 8.4.2 Amphibians as Bioindicators and Biomonitors 189 8.4.3 Suggested Amphibian Model Species for Biomonitoring the Pampa Region of Argentina 190 8.5 Description and Life Histories of Model Amphibian Species for the Pampa Region of Argentina 192 8.5.1 Leptodactylus latinasus (Jiménez de la Espada, 1875) 192 8.5.2 Leptodactylus latrans (Steffen, 1815) 193 8.5.3 Hypsiboas pulchellus (Duméril and Bibron, 1841) 195 8.5.4 Rhinella dorbignyi (Dumeril and Bibron, 1841) and Rhinella fernandezae (Gallardo, 1957) 196 8.5.5 Rhinella arenarum (Hensel, 1867) 197 8.6 Previous Biomonitoring Studies Conducted with Proposed Amphibian Model Species 199 8.6.1 Studies Using Model Species as Bioindicators 199 8.6.2 Studies Using Model Species as Biomonitors 200 References 203 Chapter 9 Odontophrynus cordobae (Anura, Cycloramphidae): A Suitable Model for Genotoxicity in Environmental Monitoring Studies 214 9.1 Biomarkers as a Tool to Assess the Impact of Environmental Contamination 214 9.2 Amphibians are Suitable Organisms to Evaluate the Genotoxic Effects of Environmental Contaminants 216 9.3 Relevant Features of Odontophrynus cordobae for Genotoxicity Studies in Environmental Monitoring 217 9.4 Conclusions 221 References 223 Chapter 10 The Direct-developing Frog Eleutherodactylus johnstonei (Eleutherodactylidae) as a Biological Model for the Study of Toxic, Cytotoxic, and Genotoxic Effects of Agrochemicals 230 10.1 Introduction 230 10.2 Natural History of the Antillean Coqui 231 10.3 Geographic Distribution 232 10.4 The Antillean Coqui as an Invasive Species 234 10.5 Conservation Status and Concerns 234 10.6 The Antillean Coqui as a Model in Ecotoxicology 235 10.7 Collection, Maintenance, and Reproduction in Captivity 237 10.7.1 Collection and Sex Determination 237 10.7.2 Taxonomic Identification 237 10.7.3 Maintenance and Reproduction in Captivity 238 10.7.4 Handling Embryos 239 10.8 Applications for Testing Environmental Xenobiotics 239 Acknowledgments 241 References 242 Chapter 11 The Lizard Salvator merianae (Squamata, Teiidae) as a Valid Indicator in Toxicological Studies 247 11.1 Introduction 247 11.2 Evaluation of Effects of Environmental Agent 251 11.3 A Pathway to the Truth 251 11.4 Goals of Biological Monitoring 253 11.5 Studies In Ovo 255 11.6 Studies In Vivo under Controlled Conditions 260 11.7 Genotoxic Evaluation of Tegu Lizard Environmentally Exposed to Pesticides 262 References 264 Chapter 12 The Terrestrial Lizard Podarcis sicula as Experimental Model in Emerging Pollutants Evaluation 271 12.1 Introduction 271 12.2 Who is the Lizard Podarcis sicula? 272 12.3 Pollution by Organic Contaminants with Estrogen-like Action: Fertilizers and Manure 273 12.4 Pollution by Heavy Metals: Cadmium 275 12.5 Podarcis sicula as Sentinel Lizard 275 12.6 Soil Pollution by Estrogen-like Substances 276 12.7 Soil Pollution by Pesticides 278 12.8 Soil Pollution by Cadmium 279 12.9 Conclusions 281 References 281 Chapter 13 The Yellow-legged Gull Larus michahellis (Charadriiformes, Laridae) as a Model Species in Ecotoxicology: Application in Monitoring and Toxicity Assessment of Environmental Pollutants 288 13.1 Introduction 288 13.2 Materials and Methods 292 13.2.1 Study Area 292 13.2.2 In Ovo PFOS Manipulation 292 13.2.3 PFOS Determination in Yolk Sac from Control Eggs 293 13.2.4 Oxidative and Genetic Biomarker Methods 294 13.2.5 Statistical Analysis 295 13.3 Results and Discussion 295 13.3.1 PFOS Concentrations in Control Eggs 295 13.3.2 PFOS Effects on Embryo Development and Morphometric Traits 296 13.3.3 PFOS Effect on Oxidative Stress and Genetic Biomarkers 297 13.4 Conclusions 301 Acknowledgments 302 References 302 Chapter 14 South American Cowbirds as Avian Models for Environmental Toxicity Testing 308 14.1 Introduction 308 14.2 Actual and Historical Use of Birds in Science and Regulatory Toxicology 309 14.2.1 Birds as Animal Models in Toxicology and Scientific Research 309 14.2.2 Avian Models in Regulatory Environmental Toxicity Testing 310 14.2.3 Pesticide Registration and Avian Toxicity Testing in South America 311 14.3 South American Cowbirds' Diversity, Distribution and Life History 312 14.3.1 Shiny Cowbird 313 14.3.2 Bay-winged Cowbird 314 14.3.3 Screaming Cowbird 315 14.4 Cowbirds as an Avian Model for Environmental Toxicity Testing 316 14.5 Methods for Maintaining and Using Cowbirds in the Laboratory for Environmental Toxicity Testing 317 14.5.1 Capture and Transport 317 14.5.2 Housing, Acclimation and Feeding 318 14.5.3 Acute Oral Toxicity Testing 319 References 320 Chapter 15 Epilogue and Final Remarks 326 Subject Index 333 Cover......Page 1 Contents......Page 12 Preface......Page 8 Section I: Terrestrial Invertebrates as Experimental Models......Page 20 1.1 Soil Invertebrates......Page 22 1.2 The Use of Invertebrates in Soil Ecotoxicology......Page 25 1.3.1 Earthworms......Page 32 1.3.2 Collembolans......Page 35 1.3.3 Enchytraeids......Page 36 1.3.4 Isopods......Page 37 Acknowledgments......Page 38 References......Page 39 2.1 Introduction......Page 50 2.2.1 Structure and Function of Soils and Soil Organism Communities......Page 52 2.3.1 Status Quo......Page 54 2.3.2 Transition......Page 55 2.3.3 New Developments......Page 56 2.3.4 Challenges......Page 57 2.3.5 Future Demands......Page 58 2.4.1 Ontology and History of Test Systems......Page 59 2.4.2 Methodological Challenges of Multispecies Tests......Page 62 2.5 Exposure of Soil Organisms Resulting From the Fate of Pesticides......Page 65 2.6.1 Specific Protection Goals......Page 67 2.6.2 Derivation of Assessment Factors......Page 68 2.6.3 TME as Surrogate Reference Tier......Page 69 References......Page 70 3.1 Introduction......Page 78 3.2.3 Exposure Following Amendment with Differing Pesticides or B[a]P......Page 81 3.2.5 The Alkaline Single Cell-gel Electrophoresis (‘comet') Assay......Page 82 3.3 Results and Discussion......Page 83 3.3.1 Comet Generation from Differing Compounds......Page 85 3.3.2 Changes in Comet Formation Following Exposure to Aged Cypermethrin Residues......Page 87 3.4 Conclusion......Page 90 References......Page 91 4.1 Introduction......Page 95 4.2.1 Heavy Metal......Page 97 4.2.2 Organic Contaminants......Page 98 4.3 Helix aspersa for Biomonitoring of Contaminated Soil......Page 99 4.4 Genotoxicity Tests with H. aspersa and Contribution to Environmental Research......Page 103 4.5 Conclusions......Page 107 References......Page 108 5.1 Introduction......Page 115 5.2 Spiders in Ecosystems Contaminated with Heavy Metals......Page 117 5.2.1 Cellular Defence Reactions in Spiders from Areas Affected by Industrial Pollution......Page 120 5.3 Spider Sensitivity to Pesticides......Page 126 5.3.1 Changes in AChE Activity......Page 128 5.3.2 Enzymatic Detoxifying Reactions......Page 129 5.3.3 Genotoxic and Cytotoxic Effects of Plant Protection Agents in Spiders......Page 130 5.4 Starvation Stress......Page 133 5.5 Conclusions......Page 134 References......Page 135 Section II: Terrestrial Vertebrates as Experimental Models......Page 142 6.1 Color in Animals......Page 144 6.2 Internal Melanin-pigmented Cells......Page 147 6.4 Response of Cutaneous Melanocytes to Aquatic Contaminants......Page 148 6.5 Response of Internal Melanocytes to Aquatic Contaminants......Page 149 6.6 Response of Melanomacrophages to Aquatic Contaminants......Page 153 References......Page 157 7.1 Introduction......Page 162 7.2 Toxicological Studies on the Impact of Pesticides on Terrestrial Life-stages of European Amphibians......Page 164 7.3 Risk Assessments for Terrestrial Life-stages of Amphibians in Pesticide Approval......Page 168 7.3.1 Surrogate Species for Terrestrial Life-stages of Amphibians......Page 169 7.4 Pesticide Formulations—Toxicity in the Mix?......Page 174 7.5 Conclusions......Page 175 References......Page 176 8.1 Introduction......Page 182 8.2.1 Amphibian Diversity and Life History......Page 183 8.2.2 Amphibian Declines......Page 184 8.3.1 Location, Geography and Characteristics......Page 185 8.3.2 Evolution of Agricultural Practices and Environmental Impacts......Page 186 8.4.1 Agriculture and Amphibian Declines......Page 188 8.4.2 Amphibians as Bioindicators and Biomonitors......Page 189 8.4.3 Suggested Amphibian Model Species for Biomonitoring the Pampa Region of Argentina......Page 190 8.5.1 Leptodactylus latinasus (Jiménez de la Espada, 1875)......Page 192 8.5.2 Leptodactylus latrans (Steffen, 1815)......Page 193 8.5.3 Hypsiboas pulchellus (Duméril and Bibron, 1841)......Page 195 8.5.4 Rhinella dorbignyi (Dumeril and Bibron, 1841) and Rhinella fernandezae (Gallardo, 1957)......Page 196 8.5.5 Rhinella arenarum (Hensel, 1867)......Page 197 8.6.1 Studies Using Model Species as Bioindicators......Page 199 8.6.2 Studies Using Model Species as Biomonitors......Page 200 References......Page 203 9.1 Biomarkers as a Tool to Assess the Impact of Environmental Contamination......Page 214 9.2 Amphibians are Suitable Organisms to Evaluate the Genotoxic Effects of Environmental Contaminants......Page 216 9.3 Relevant Features of Odontophrynus cordobae for Genotoxicity Studies in Environmental Monitoring......Page 217 9.4 Conclusions......Page 221 References......Page 223 10.1 Introduction......Page 230 10.2 Natural History of the Antillean Coqui......Page 231 10.3 Geographic Distribution......Page 232 10.5 Conservation Status and Concerns......Page 234 10.6 The Antillean Coqui as a Model in Ecotoxicology......Page 235 10.7.2 Taxonomic Identification......Page 237 10.7.3 Maintenance and Reproduction in Captivity......Page 238 10.8 Applications for Testing Environmental Xenobiotics......Page 239 Acknowledgments......Page 241 References......Page 242 11.1 Introduction......Page 247 11.3 A Pathway to the Truth......Page 251 11.4 Goals of Biological Monitoring......Page 253 11.5 Studies In Ovo......Page 255 11.6 Studies In Vivo under Controlled Conditions......Page 260 11.7 Genotoxic Evaluation of Tegu Lizard Environmentally Exposed to Pesticides......Page 262 References......Page 264 12.1 Introduction......Page 271 12.2 Who is the Lizard Podarcis sicula?......Page 272 12.3 Pollution by Organic Contaminants with Estrogen-like Action: Fertilizers and Manure......Page 273 12.5 Podarcis sicula as Sentinel Lizard......Page 275 12.6 Soil Pollution by Estrogen-like Substances......Page 276 12.7 Soil Pollution by Pesticides......Page 278 12.8 Soil Pollution by Cadmium......Page 279 References......Page 281 13.1 Introduction......Page 288 13.2.2 In Ovo PFOS Manipulation......Page 292 13.2.3 PFOS Determination in Yolk Sac from Control Eggs......Page 293 13.2.4 Oxidative and Genetic Biomarker Methods......Page 294 13.3.1 PFOS Concentrations in Control Eggs......Page 295 13.3.2 PFOS Effects on Embryo Development and Morphometric Traits......Page 296 13.3.3 PFOS Effect on Oxidative Stress and Genetic Biomarkers......Page 297 13.4 Conclusions......Page 301 References......Page 302 14.1 Introduction......Page 308 14.2.1 Birds as Animal Models in Toxicology and Scientific Research......Page 309 14.2.2 Avian Models in Regulatory Environmental Toxicity Testing......Page 310 14.2.3 Pesticide Registration and Avian Toxicity Testing in South America......Page 311 14.3 South American Cowbirds' Diversity, Distribution and Life History......Page 312 14.3.1 Shiny Cowbird......Page 313 14.3.2 Bay-winged Cowbird......Page 314 14.3.3 Screaming Cowbird......Page 315 14.4 Cowbirds as an Avian Model for Environmental Toxicity Testing......Page 316 14.5.1 Capture and Transport......Page 317 14.5.2 Housing, Acclimation and Feeding......Page 318 14.5.3 Acute Oral Toxicity Testing......Page 319 References......Page 320 Chapter 15 Epilogue and Final Remarks......Page 326 Subject Index......Page 333 The potential impact of anthropogenic pollutants such as agrochemicals on the environment is of global concern. Increasing use of certain compounds can result in contamination of food, water and atmospheric systems and in order to combat this pollution it is important to be able to accurately monitor the short and long term effects. This book describes the latest non-traditional terrestrial species models used as indicators of the toxic effects of environmental pollutants. The book enables understanding of the effects of pollutants in non-target species, and therefore enables analysis of the effects on ecosystems. This book will be of interest to anyone interested in developing new biomarker species with high degrees of ecological relevance. It will serve as a useful resource for regulatory and research toxicologists, particularly those interested in soil screening and the effects of pollutants on wildlife and insects and their use as biological indicators As part of the OECD's Pesticide Programme, a Workshop on Avian Toxicity Testing was held in Pensacola, Florida, on 4-7 December 1994. It was jointly organised by the Society of Environmental Toxicology and Chemistry (SETAC) and the OECD. This Workshop report considers the fundamental reasons information about the toxicity of chemicals to birds is needed, and thereby identify the critical features which are essential for the tests used to generate this information; evaluates the positive and negative features of existing test methods, and of proposed alternatives, against the required critical features; develops proposals for the revision and development of OECD Test Guidelines for avian toxicity and avoidance tests, as appropriate; and develops a framework to govern the way these tests are requested and used in practice in risk assessment. Executive Summary 1. Introduction 2. Testing for Acute Toxicity 3. Testing for Dietary Toxicity 4. Testing for Effects on Reproduction 5. Testing for Avoidance 6. Framework for the Use of Avian Tests in the Risk Assessment of Pesticides 7. Conclusions and RecommendationsAcknowledgements Annex 1 Workshop Participants Annex 2 Questionnaire: Request for Information and Opinions from Participants Annex 3 Summary of Responses to the Pre-Workshop Questionnaire: General Issues Annex 4 Comparison of Existing Avian Toxicity and Avoidance Tests Annex 5 Reading List OECD Environmental Health and Safety Publications