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Appraisal of Test Methods for Sex-Hormone Disrupting Chemicals Capable of Affecting the Reproductive Process

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Appraisal of Test Methods for Sex-Hormone Disrupting Chemicals Capable of Affecting the Reproductive Process APPRAISAL OF TEST METHODS FOR SEX-HORMONE DISRUPTING CHEMICALS CAPABLE OF AFFECTING THE REPRODUCTIVE PROCESS Prepared by the MRC Institute for Environment and Health for the UK Department of Environment, Transport and the Regions 1998 Prepared by the MRC Institute for Environment and Health Written by: Mr. P. Holmes Dr. C Humfrey Mr. M. Scullion With additional material from : Dr M Taylor 2 FOREWORD Over a number of years anecdotal observations and scientific evidence have come to light which suggest that chemicals present in the environment may cause adverse health effects in humans and wildlife because of their capacity to mimic or influence the action of endogenous hormones such as oestrogens and androgens. Current concerns centre around the apparent increase in human and animal disease states linked to the action of sex-hormones and reported observations from human epidemiological studies of reductions in sperm counts and fertility, increases in the incidence of testicular cancer and congenital birth defects in men and increases in the incidence of breast cancer in women. In wildlife species the occurrence of gross birth deformities, behavioral abnormalities and both feminisation and masculinisation are major concerns. While there remains much scientific uncertainty about the nature and magnitude of the human and environmental risks associated with exposure to endocrine disrupting chemicals, the concerns are such that in November 1996, OECD Member countries decided to work together on international projects to: • coordinate national and regional activities on the risk assessment and management of endocrine disrupting chemicals; • develop internationally acceptable methods for the hazard characterization of endocrine disrupting chemicals; and • harmonise risk characterization approaches and regulatory approaches. This Detailed Review Paper (DRP) is intended make an important contribution to this work serving as the basis for the first step in the consideration and development of OECD guidelines for the testing of chemicals for endocrine disrupting effects. OECD Test Guidelines play a critical role in ensuring efficent and effective procedures are available to identify chemical hazards. The focus of the DRP is on test methods for sex-hormone disrupting chemicals capable of affecting the reproductive process. Other hormone systems which are also important in the control of reproduction such as the thyroid and adrenal systems are not considered. In addition, test methods for the effects of sex hormones on non- reproductive processes such as brain development and behaviour are considered to be beyond the scope of this document. Accordingly, the DRP makes an inventory of existing OECD test methods relevant to the assessment of the effects of sex hormone disrupters on the reproductive systems of humans and wildlife; describes relevant non-regulatory test methodologies used by the research community; makes a critical assessment of existing test methodologies to detect sex hormone disrupting chemicals; identifies possible enhancements to existing test methodologies; and identifies both non-regulatory test methodologies suitable for further development and outstanding research requirements. 3 It has been written to provide a state of the art review. As understanding and scientific consensus on endocrine disruption continue to rapidly emerge there will be new tests developed which may warrant future consideration and discussion. In the area of existing tests the report assesses only OECD test methodologies. Where regulatory authorities in specific countries have their own relevant test methodologies it is expected that these will be taken into account as proposals for enhancements or new OECD test methods are further discussed. Similarly, as scientific consensus on this subject develops undoubtedly there will be new perspectives and issues that need consideration. Of particular importance are effects on wildlife. In this area there have been several useful Workshops such as the Joint OECD/SETAC EMWAT Workshop on Endocrine Modulators and Wildlife Assessment and Testing held in Veldhoven, Netherlands 10-13 April 1997 (Tattersfield et al., 1997). The report on this Workshop therefore provides a useful adjunct to the present text. The Executive Summary of this Workshop is reproduced as an Annex to this Monograph for the convienience of readers. The DRP was contributed to the OECD by the Government of the United Kingdom and was prepared by the UK Medical Research Council’s Institute for Environment and Health. It was considered in draft form by representatives of OECD Member countries through the forum of the National Test Guideline Coordinators. A large number of comments on the draft were provided by Member countries, many of which have been taken into account in finalizing the report. The comments provided support the document as accurate, comprehensive and well balanced and a good basis for the discussion of relevant Test Guidelines. Initially the Detailed Report Paper will be considered in detail by a Working group on Endocrine Disrupter Testing and Assessment. This Working Group will consider the conclusions and recommendations of the Detailed Review Paper and make recommendations to the OECD for further work. The Joint Meeting of the Chemicals Group and the Management Committee of the Special Programme on the Control of Chemicals have recommended that the report be derestricted and published as an OECD Monograph. It has been made public under the responsibility of the Secretary-General of the OECD. 4 CONTENTS PAGE 1. EXECUTIVE SUMMARY ................................................................................................................ 7 1.1 LIMITATIONS OF CURRENT OECD TEST METHODS .............................................................. 7 1.2 MODIFICATIONS TO EXISTING OECD TEST METHODS ........................................................ 8 1.3 NON-REGULATORY TEST MODELS PROPOSED FOR FURTHER DEVELOPMENT/ADOPTION ............................................................................................................... 8 1.4 REQUIREMENTS FOR BASIC RESEARCH .................................................................................. 9 2. INTRODUCTION............................................................................................................................ 11 2.1 GENERAL BACKGROUND .......................................................................................................... 11 2.2 REVIEW OBJECTIVES AND PROCEDURES ............................................................................. 12 2.3 SEX DETERMINATION AND REPRODUCTIVE CONTROL SYSTEMS IN THE ANIMAL KINGDOM ............................................................................................................................................ 13 2.3.1 GENOTYPIC SEX DETERMINATION .................................................................................... 13 2.3.2 SEX DIFFERENTIATION AND HORMONAL CONTROL IN MAMMALS ............................. 14 2.3.3 SEX DIFFERENTIATION AND HORMONAL CONTROL IN NON-MAMMALIAN VERTEBRATES.................................................................................................................................. 16 2.3.3.1 BIRDS .................................................................................................................................................16 2.3.3.2 REPTILES AND AMPHIBIANS........................................................................................................16 2.3.3.3 FISH ....................................................................................................................................................17 2.3.4 SEX DETERMINATION AND DIFFERENTIATION AND HORMONAL CONTROL IN INVERTEBRATES.............................................................................................................................. 17 2.3.4.1 ANNELIDA ........................................................................................................................................18 2.3.4.2 MOLLUSCA .......................................................................................................................................19 2.3.4.3 CRUSTACEA .....................................................................................................................................19 2.3.4.4 INSECTA ............................................................................................................................................20 2.4 MECHANISMS OF SEX HORMONE DISRUPTION ................................................................... 20 3. OVERVIEW OF EXISTING REGULATORY TEST METHODS............................................. 22 3.1 REGULATORY BACKGROUND.................................................................................................. 22 3.2 OECD ACTIVITIES ........................................................................................................................ 22 3.3 REVIEW OF OECD GUIDELINES................................................................................................ 37 3.3.1 MAMMALIAN STUDY DESIGNS ............................................................................................ 37 3.3.2 NON-MAMMALIAN STUDY DESIGNS................................................................................... 40 3.4 POTENTIAL ENHANCEMENTS TO EXISTING OECD GUIDELINES .................................... 41 4. CRITICAL ASSESSMENT OF NON-REGULATORY
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