See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/6223709 Ring-Testing and Field-Validation of a Terrestrial Model Ecosystem (TME) - An Instrument for Testing Potentially Harmful Substances: Effects of Carbendazim on Earthworms Article in Ecotoxicology · February 2004 DOI: 10.1023/B:ECTX.0000012408.58017.08 · Source: PubMed CITATIONS READS 31 116 6 authors, including: Cornelis A M van Gestel Thomas Moser Vrije Universiteit Amsterdam Eurofins Agroscience Germany 456 PUBLICATIONS 16,200 CITATIONS 30 PUBLICATIONS 447 CITATIONS SEE PROFILE SEE PROFILE Some of the authors of this publication are also working on these related projects: Endocrine disruption in the terrestrial isopod Porcellio scaber View project Effects on heavy metals Earthworms View project All content following this page was uploaded by Cornelis A M van Gestel on 17 May 2014. The user has requested enhancement of the downloaded file. Ecotoxicology, 13, 105–118, 2004 Ó 2004 Kluwer Academic Publishers. Manufactured in The Netherlands. Ring-Testing and Field-Validation of a Terrestrial Model Ecosystem (TME) – An Instrument for Testing Potentially Harmful Substances: Effects of Carbendazim on Earthworms JO¨ RG RO¨ MBKE,1,* CORNELIS A.M. VAN GESTEL,2 SUSAN E. JONES,3 JOSE´ E E. KOOLHAAS,2 JOSE´ M.L. RODRIGUES4 AND THOMAS MOSER1 1ECT Oekotoxikologie GmbH, Bo¨ttgerstr. 2-14, D-65439 Flo¨rsheim am Main, Germany 2Institute of Ecological Science, Vrije Universiteit, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands 3University of Wales, School of Agricultural and Forestry Sciences, Bangor, Gwynedd LL57 2UW, Wales, UK 4Departamento de Biologia, Universidade de Aveiro, Aveiro, Portugal Accepted 24 October 2002 Abstract. The effects of the fungicide carbendazim (applied in the formulation DerosalÒ) on earthworms (Lumbricidae) was determined in Terrestrial Model Ecosystem (TME) tests and field-validation studies. TMEs consisted of intact soil columns (diameter 17.5 cm; length 40 cm) taken from a grassland or, in one case, from an arable site. The TMEs were taken from the same site where the respective field-validation study was performed. The tests were performed in Amsterdam (The Netherlands), Bangor (Wales, UK), Coimbra (Portugal) and Flo¨ rsheim (Germany). The sites selected had an earthworm coenosis representative of the different land use types and regions. In addition, the differences between the coenosis found in the TMEs and the respective field sites were in general low. A high variability was found between the replicate samples, which reduces the probability of determining significant differences by the statistical evaluation of the data. Similar effects of the chemical treatment were observed on abundance as well as on biomass. Effects were most pronounced 16 weeks after application of the test chemical. The observed effects on earthworm abundance and biomass did not differ between the TME tests and the respective field-validation studies. Effects on earthworm diversity were difficult to assess since the number of individuals per species was low in general. However, the genus Lumbricus and in particular L. terrestris and L. rubellus seemed to be more affected by the chemical treatment than others. NOEC and EC50-values derived from the TME pre-test, the TME ring-test and the field-validation study indicate that the TMEs of the different partners delivered comparable results although different soils were used. Due to the high variability NOECs could often not be determined. The EC50-values for the effect of carbendazim on earthworm abundance ranged between 2.04 and 48.8 kg a.i./ha (2.71–65.2 mg/kg soil) and on earthworm biomass from 1.02 to 34.6 kg a.i./ha (1.36–46.0 mg/kg soil). These results indicate that the abundance and biomass of earth- worms are suitable endpoints in ecotoxicological studies with TMEs. Keywords: carbendazim; Lumbricidae; earthworms; soil mesocosms; community effects *To whom correspondence should be addressed: Tel.: +49-6145-9564-20; Fax: +49-6145-9564-99; E-mail: [email protected] 106 Ro¨mbke et al. Introduction throughout Europe (Cuppen et al., 2000; Framp- ton and Wratten, 2000). This chemical was also Risk assessment of chemicals is usually based on chosen as a reference substance in the earthworm the results of single species toxicity tests, which are chronic reproduction test (ISO 1998; OECD 2001), performed in the laboratory (Van Leeuwen and since it is known for its high toxicity to earth- Hermens, 1995). It is, however, realised that such worms. single species tests may not be sufficient to predict effects on the level of ecosystem structure and functioning (Cairns, 1984). In the past, such effects Materials and methods have been studied in the field but the results are difficult to assess and the efforts are high. There- Experimental set-up fore, mesocosm tests have been advocated, which may close the gap between laboratory and field Three types of tests were performed: in the first studies. For the soil environment, this has resulted year a TME pre-test, in the second year, based on in the development of several types of model eco- the experience gained during the pre-test, the TME systems (Morgan and Knacker, 1994; Sheppard, ring-test and in parallel to the ring-test a field- 1997; Edwards et al., 1998; Knacker et al., 2004). validation study. The TME-project was conducted Such model ecosystems may be applied in a higher at four sites throughout Europe by the following tier of risk assessment, when results of single spe- project partners: ECT Oekotoxikologie GmbH, cies laboratory toxicity tests provide reasons for Flo¨ rsheim, Germany (1), Vrije Universiteit Am- concern with regard to the potential risk of a sterdam (Institute of Ecological Science, The chemical in the soil environment (see Weyers et al., Netherlands) (2), University of Wales, Bangor 2004). (School of Agricultural and Forestry Sciences, In this paper, results on earthworm abundance, UK) (3) and Universidade de Coimbra (Instituto biomass and diversity gained by tests using Ter- Ambiente e Vida, Portugal) (4). In Coimbra, TME restrial Model Ecosystems (TMEs) are described. tests and the field-validation study were performed TMEs are defined as controlled, reproducible at an arable site, whereas the respective work at systems that attempt to simulate the processes and the three other sites was done on a grassland. The interactions of components in a portion of the properties of the soils from these four sites were terrestrial environment (Gillett and Witt, 1980; measured using ISO guidelines (ISO 1993; ISO Sheppard, 1997). These tests were performed 1994; ISO 1995; Table 1). The TME tests started within the framework of the TME-project spon- with the extraction of the TME soil cores. TMEs sored by the European Union (Contract No.: were taken by means of a soil core extractor, ENV4-CT97-0470). The aim of this project was containing a HDPE tube (diameter 17.5 cm; the improvement and validation of the TME test length 40 cm), which served as a soil core encase- system, first described by Van Voris et al. (1985) ment. Grass was cut just before TME extraction. and used in studies reported by Knacker et al. The TMEs were either placed in temperature- (1989), Frederickson et al. (1991) and Chekai et al. controlled carts in a climatic chamber (Amster- (1993). For that purpose, the TME tests were dam, Flo¨ rsheim, Coimbra) or in a greenhouse performed at four different European sites with (Bangor). TMEs were irrigated up to three times different soils, but using similar equipment, test per week using artificial rainwater slightly modi- chemical, test design, and endpoints. The tests fied according to Velthorst (1993). The model were designed in a way which should allow a chemical carbendazim was applied after an accli- comparison of NOEC- and EC50-values for several matisation period of two to four weeks. endpoints. In order to investigate the ecological For the field-validation study, 30 field plots, realism of this laboratory TME test system, a field- each 25 m2, were marked out by each partner at validation study was performed. Carbendazim was the same site where the TMEs were extracted. Six chosen as the model chemical for the TME tests plots served as controls, and 24 plots were treated and the field-validation study. Carbendazim is a with the model chemical (four plots for each of the fungicide that is used on a large scale in agriculture six treatment levels; the plots were completely TME – Effects of Carbendazim on Earthworms 107 Table 1. Soil properties and site use of the four experimental fields from which the soil cores for the TME tests were extracted and the respective field-validation studies were performed (for details see Knacker et al. (2004); OM = organic matter Participant/ Country code Texture Clay (%)OM(%) pH (KCl) Land use location Amsterdam NL Silty loam 7.9 4.5 4.8–5.1 Meadow Bangor UK Loam 20.5 6.1 5.8–6.6 Pasture Coimbra P Silty loam 24.7 3.4 6.4–7.1 Arable field Flo¨ rsheim D Silty clay loam 36.5 5.2 5.3–5.9 Meadow randomised). Before spraying the model chemical tailed description of the set-up of the TME- the grass cover was cut on the grassland sites project, see Knacker et al. (2004). (Amsterdam, Bangor, Flo¨ rsheim) or the soil was ploughed at the arable site (Coimbra). Earthworm sampling Carbendazim was applied to the TMEs and field plots as DerosalÒ, containing 360 g carbendazim/l. In the TME pre-test, earthworm samples were In the TME pre-tests, carbendazim was applied at taken 1, 8 and 16 weeks after application. After dosages of 0 (T0), 0.36 (T1), 2.16 (T2), 13.0 (T3) taking the samples for all other endpoints, the re- and 77.8 (T4) kg a.i./ha.