Validating the enchytraeid reproduction test: organisation and results of an international ringtest

In this paper the experiences concerning the organisation and results of the enchytraeid reproduction test (ERT) ringtest are summarised (for details see J. Römbke, T. Moser, Organisation and Performance of an International Ringtest for the Validation of the Enchytraeid Reproduction Test, vols. I and II. UBA-Texte 4/99, 1999, 150, 223 pp). The performance of this ringtest was in line with requirements published by OECD. It was sponsored by the German Federal Environmental Agency (Umweltbundesamt; UBA, Berlin). The UBA was also actively engaged (together with the European Chemicals Bureau, Ispra) in a scientific task force, which consisted of seven scientists experienced in terrestrial ecology and ecotoxicology. 29 institutions (mainly from universities and contract research laboratories) from 15 countries of Europe and North America participated actively in the ringtest. The co-ordinating laboratory centrally distributed the test chemicals, guidance papers and test organisms to all participants. In addition, several training courses were organised. Most participants performed two tests with the fungicide Carbendazim and two with 4-nitrophenol. For each chemical, one test was designed according to an ECx approach and the other according to an NOEC approach. Several aspects of the test protocol were modified based on the experiences gained during the ringtest. A major aspect of the project was the detailed statistical evaluation of the test results (for details see A. Weyers, J. Römbke, T. Moser, T. Ratte, Results of and statistical implications from the enchytraeid reproduction ringtest, 2001, submitted), leading to recommendations for an optimised ecotoxicological test design. A total of 92 tests were performed according to the protocol which is among the highest number ever performed in a ringtest. About 72% met the validity criteria (control mortality < 20%, > 25 juveniles per 10 adults). The effects of the two test chemicals on enchytraeid reproduction were in the range expected from data in the literature on oligochaete toxicity. However, statistical evaluation of the data was sometimes difficult as a result of the high variability in the number of juveniles. This variability was caused by several reasons, including hormesis effects or lack of experience of some participants. Comparison of the data from the NOEC and ECx approaches pointed to a clear advantage in favour of the latter. In most cases EC10 values were lower than the NOEC values determined in the same test. For details see A. Weyers et al. (loc. cit.). Reproducibility of the test data and practicability of the ERT ringtest were of the same order of magnitude as other ringtests recently performed. As a result of the ringtest, the ERT draft guideline was significantly improved. The new version is currently being standardised by OECD, ISO and ASTM.