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Validation Report – Results of an International Ring Test According to the Draft Guideline Predatory Mite Reproduction Test I

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Validation Report – Results of an International Ring Test According to the Draft Guideline Predatory Mite Reproduction Test I Validation report – Results of an International Ring test According to the Draft Guideline Predatory mite reproduction test in soil (Hypoaspis (Geolaelaps) aculeifer) Compiled by J. Römbke, L. Becker, B. Dark, Th. Moser, N. Halsall, W. Powley, A. Ruf, C. Scholer, E. Smit, P. Wege, N. Zenz On behalf of the HASTE (Hypoaspis Aculeifer Soil TEst ) Group June 08, 2007 1 Content: 0. Executive Summary 1. Introduction 1.1 Background 1.2 Information on the ring test 1.3 Work performed in the HASTE Ring test 2. Methodological Overview 2.1 Compilation of experiences made with the standard tests 2.2 Results from the non-standardized tests 3. Summary of Results 3.1 Experimental design 3.2 Summary of control data 3.3 Method of analysis and results 3.4 Discussion 4. References Annex I: Information on the HASTE group Annex II: Draft Test Guideline (ninth draft; April 23, 2007) available on the OECD public website: [http://www.oecd.org/document/62/0%2C3343%2Cen_2649_34377_2348862_1_1_1_1%2C00.html] Annex III: Ring Test Report - Statistical Analysis (final version; May 14, 2007) available on the OECD public website: [http://www.oecd.org/document/62/0%2C3343%2Cen_2649_34377_2348862_1_1_1_1%2C00.html] 2 0. Executive Summary A new Test Guideline has been developed, which is designed to be used for assessing the effects of chemical substances in soil on the reproductive output of the soil mite species Hypoaspis (Geolaelaps) aculeifer Canestrini (Acari: Laelapidae). H. aculeifer represents an additional trophic level to the species for which guidelines are already available. The main endpoint is the reproduction of the mites without discrimination and quantification of the different stages of the reproductive cycle. Based on already standardised OECD and ISO test guidelines as well as ideas from literature, the ad-hoc working group HASTE (Hypoaspis aculeifer Soil Test) prepared a draft guideline using OECD format in 2005. This group has about 30 members from different countries, representing authorities, industry (including contract laboratories) and universities. Using the draft guideline as a starting point, HASTE developed a study protocol which was afterwards used in an international ring test. Twelve laboratories performed in total 48 tests using two test chemicals (dimethoate and boric acid) and two test designs (NOEC, ECx). Only five tests were not valid. Seven tests (plus further work) were intended to clarify methodological details, in particular to determine the optimal test duration. Originally, a test duration of 16 days was proposed but finally 14 days were determined to be more suitable and the results are at least as sensitive. The change of the test duration was the only relevant change of the test method caused by the ring test experiences. The results determined in the ring test proved the suitability of the proposed test method. The LC50 values for both test chemicals differed by less than a factor of 2.5 from the mean and no statistically significant differences were found between laboratories.The EC50 values from the dimethoate tests differed by less than a factor of two from the mean and no statistically significant differences were found between laboratories. In the case of boric acid the range of EC50 values was broader (they differed by less than a factor of five from the mean), but still there were no statistically significant differences between laboratories. Smaller treatment effects could be detected in the tests with higher numbers of replicates. After the discussion of the results of the ring test in a HASTE workshop (January 2007), the group decided to recommend to include the final draft version of this test guideline into the OECD test guideline program. This recommendation is based on the evidence summarised in this report and in particular its Annexes. 1. Introduction 1.1 Background The proposed test guideline is intended to be used for the environmental risk assessment of chemical substances, in particular plant protection products (PPPs). The detailed requirements for this test are laid down in the Terrestrial Guidance Document (EC 2002). However, the test is also suitable for other chemical substances. Its applicability is supported by the fact that ecotoxicological studies using the same and very closely related mite species as proposed here are known in the literature for many years (e.g. Schlosser & Riepert 1992). Finally, proposals for standardised tests with Hypoaspis aculeifer have already been made (Schlosser & Riepert 1991/92; Krogh & Axelsson 1998; Bakker et al. 2003). The test is designed to estimate the effects of a test chemical on the reproductive output of the predatory soil mite Hypoaspis (Geolaelaps) aculeifer. The mites are exposed under controlled conditions to the chemical which has been mixed into the test substrate (usually OECD artificial soil). H. aculeifer is considered to be a relevant representative of soil fauna and predatory mites in particular for the following reasons: 3 - It is worldwide distributed (Karg 1993). - The ecology of this species is well-known (e.g. Ruf 1991; 1995; 1996; Heckmann et al. 2007) - Ecotoxicological background information is also available (e.g. Bakker et al. 2003; Heckmann et al. 2005) - In the laboratory, it is easy to culture and to handle. Therefore, this species was proposed as part of a battery of standard soil tests (Løkke et al. 2002), since it represents an additional tropic level to the species for which guidelines are already available. The proposed draft guideline has been developed by the HASTE (Hypoaspis aculeifer Soil TEst) group. The group was founded at a meeting at the BBA in Braunschweig (Germany) on January 31, 2003. Since then, the group met once or twice per year in order to discuss various versions of the draft guideline. A first draft, already written in OECD format, was developed in late 2004. The practical work of the ring test started in late 2005, using the sixth version as its basis. It was supported by the German Environmental Agency (UBA, Dessau; S. Schmitz) and the Dutch National Institute for Public Health and Environmental Protection (RIVM, Bilthoven; E. Smit). During a final meeting of the HASTE group in Bilthoven (The Netherlands) the results of the ring test were discussed and a new version (already the ninth one) of the guideline ready to be submitted to OECD was agreed-on. Already in late 2005, The Netherlands agreed to act as a lead country for this new guideline within OECD. Afterwards, encouraging comments from various member countries were received. This document will be used for another commenting round. It consists of four parts: - A summary report, presenting the main methodological experiences from the ring test as well as the most important results of the this exercise - Some information about the HASTE group in Annex I; - The newest (ninth) version of the draft guideline, providing all the details how to perform the test, as Annex II - In a third Annex, a detailed listing of the individual test results including their statistical evaluation is provided. 1.2 Information on the ring test In theory, in ecotoxicology a new test is developed after a specific requirement has been identified by authorities. In reality, test ideas were often developed as part of a study with other aims (e.g. basic ecological investigations on stress reactions). Using such information as a starting point, the idea has to be transformed into a standardised test method, usually in one laboratory followed by an inter-laboratory comparison study with the aim to produce a draft test guideline. The final step of this process would be the performance of an international ring test in order to validate the method. All details are published by the responsible standardisation organisation (here: OECD (2005a)). The HASTE group had 32 members from seven countries and with very different background, i.e. governmental institutions, universities, chemical industry and contract laboratories. Its Organizing Committee / Advisory council consist of nine persons, representing the main institutional groups as well as countries. 4 In total, 12 laboratories participated in the ring test. Most of them (eight) were contract laboratories while two belonged to the chemical industry. One public institution as well as one university did also participate. About two third (seven) of the ring test participants are located in Germany. The regional background of the remaining institutions is as follows: United Kingdom (two), Denmark (one), The Netherlands (one) and Switzerland (one). In the following, names and addresses of the HASTE ring test participants are listed. Andrea Ruf University of Bremen, FB 2 UFT, Department of Ecology Postbox 330440, D- 28334 Bremen. E-mail: [email protected] Frank Bakker: MITOX, Trial Management BV, Kruislaan 406, 1098SM Amsterdam, The Netherlands. E-mail: [email protected] Fred Heimbach: Bayer AG, Institute for Environmental Biology, Crop Protection Department, D- 51368 Leverkusen, Germany. E-mail: [email protected] Karin Nienstedt: SpringbornSmithers Laboratories (Europe) AG, Terrestrial Ecotoxicoloy, Seestr. 21, CH-9326 Horn, Switzerland. E-mail: [email protected] Lennart Schulz: BioChem agrar GmbH, Kupferstr. 6, D-04827 Machern OT Gerichshain, Germany. E-mail: [email protected] Nicola Davies: CEM Analytical Services Ltd., Glendale Park, Fernbank Road, North Ascot, Berkshire, SL5 8JB, UK. E-mail: [email protected] Paul-Henning Krogh: Dept. of Terrestrial Ecology, National Environmental Research Institute, Vejlsǿvej 25, DK-8600 Silkeborg, Denmark. E-mail: [email protected] Stephen Vinall: Mambo-Tox Ltd, 2 Venture Road, Chilworth Science Park, Chilworth, Southampton, SO16 7NP, UK. E-mail: [email protected] Thomas Moser: ECT Oekotoxikologie GmbH, Boettgerstr. 2-14, D-65437 Floersheim, Germany. E-mail: [email protected] Ulf Lührs: IBACON GmbH, Arheilger Weg 17, D-64380 Rossdorf, Germany. E-mail: [email protected] Uschi Kölzer: GAB Biotechnologie GmbH, Eutingerstr. 24, D-75223 Niefern-Öschelbronn, Germany. E-mail: [email protected] Wolfgang Krieg: BASF AG, Crop Protection Division, Agricultural Center – APD/EE, Li 425.
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