Effects of the veterinary pharmaceutical ivermectin on soil invertebrates in laboratory tests.

As part of the risk assessment of veterinary pharmaceuticals, the potential impact of these chemicals on soil ecosystems has to be determined according to European law. However, almost no data from standardized laboratory tests are available. Therefore, in the EU FP6 ERAPharm, the effects of the widely used veterinary pharmaceutical ivermectin, an anthelminthic, were studied in chronic laboratory tests performed according to OECD (Organisation for Economic Co-operation and Development) and ISO (International Organization for Standardization) guidelines. In detail, three soil invertebrate species--the earthworm Eisenia fetida, the springtail Folsomia candida, and the predatory mite Hypoaspis aculeifer--were tested. The nominal concentrations of ivermectin mixed into the test substrate artificial soil was verified using residue analysis, which indicated that the test substance is persistent for at least up to 28 days. As expected when considering the mode of action of this substance, survival and reproduction of collembolans were clearly affected [LC(50) = 8.4 mg/kg soil dry weight (dw); NOEC(repro) = 0.3 mg/kg soil (dw)]. Predatory mites reacted less sensitively [LC(50) > or = 31.6 mg/kg soil (dw); NOEC(repro) = 3.2 mg/kg soil (dw)]. Earthworm survival and reproduction were affected in the same order of magnitude as the predatory mites [LC(50) > or = 10 mg/kg soil (dw); NOEC(repro) = 2.5 mg/kg soil (dw)]. These results are in good agreement with the few data known from nonstandardized tests for the same or related soil invertebrate species. The results of these tests indicate that the effects of ivermectin on soil invertebrates--in particular, collembolans--cannot be excluded at field-relevant concentrations, as determined in a risk assessment according to VICH guidelines. More sophisticated higher-tier tests (e.g., in multispecies or semifield test systems) are recommended in order to assess the potential risk more accurately.

Environmental risk assessment of veterinary pharmaceuticals: development of a standard laboratory test with the dung beetle Aphodius constans.

The environmental risk assessment of veterinary pharmaceuticals for dung beetles is required if the substance is an anti-parasiticide for the treatment of pasture animals. However, the demonstration of the environmental safety of those substances for dung fauna is hampered by the fact that no standardized laboratory test system is currently available. Here a test system using the temperate dung beetle species Aphodius (Agrilinus) constans (Scarabaeidae: Aphodiinae) Duftschmidt is described. The survival of first instar larvae of A. constans exposed to a model substance, dimethoate, spiked into formulated (i.e. dried, formulated and re-wetted) or fresh dung was measured over a period of three weeks. Larvae performed better in formulated dung which also proved to be more suitable for mixing-in test substances homogenously. Dimethoate caused significant larval mortality with LC50 values within a range of 1.3-2.8 mg a.s./kg dung (d.w.), depending on the dung type. Based on the data presented here, it is recommended to incorporate this new test system in the risk assessment process for veterinary pharmaceuticals. However, an international ringtest should to be performed beforehand to ensure adequate validation of the method.

Effect of different soil types on the enchytraeids Enchytraeus albidus and Enchytraeus luxuriosus using the herbicide Phenmedipham.

Soil ecotoxicology studies are usually performed in standard soils, such as the OECD (Organization for Economic Cooperation and Development) artificial soil or the LUFA St. 2.2, a natural soil. When assessing the toxic effects in the environment, soil properties are often different from those in standard soils, which might lead to a different exposure situation for the test species and, therefore, to a different evaluation of the risk of the test substance. Selected to cover a broad range of properties and based on the Euro-Soils concept, 18 different soils were studied regarding their suitability to two test species: Enchytraeus albidus and Enchytraeus luxuriosus (Enchytraeidae). In reproduction tests, the test species reacted differently to the tested soils, but both enchytraeids did not survive in acid soils (i.e. pH

The effect of tributyltin-oxide on earthworms, springtails, and plants in artificial and natural soils.

Chemical bioavailability in Organisation for Economic Co-operation and Development (OECD) artificial soil can contrast with bioavailability in natural soils and produce ecotoxicologic benchmarks that are not representative of species' exposure conditions in the field. Initially, reproduction and growth of earthworm and Collembolan species, and early seedling growth of a dicotyledonous plant species, in nine natural soils (with a wide range of physicochemical properties) and in OECD soil were evaluated. Soils that supported reproduction and growth of the test species were then used to investigate the toxicity of tributyltin-oxide (TBT-O). Natural soils caused greater toxicity of TBT-O to earthworms (EC(50) values varied from 0.5 to 4.7 mg/kg soil dry weight [dw]) compared with toxicity in OECD soil (EC(50) = 13.4 mg/kg dw). Collembolans were less sensitive to TBT-O than earthworms in natural soils, with EC(50) values ranging from 23.4 to 177.8 mg/kg dw. In contrast, the toxicity of TBT-O to collembolans in OECD soil (EC(50) = 104.0 mg/kg dw) was within the range of EC(50) values in natural soils. Phytotoxicity tests revealed even greater difference between the effects in natural soils (EC(50) values ranged from 10.7 to 189.2 mg/kg dw) and in OECD soil (EC(50) = 535.5 mg/kg dw) compared with results of the earthworm tests. Studies also showed that EC(50) values were a more robust end point compared with EC(10) values based on comparisons of coefficients of variation. These results show that toxicity testing should include studies with natural soils in addition to OECD soil to better reflect exposure conditions in the field.

Effects of different soil types on the Collembolans Folsomia candida and Hypogastrura assimilis using the herbicide Phenmedipham.

Soil ecotoxicology studies are usually performed in standard soils such as Organization for Economic Cooperation and Development artificial soil or LUFA ST. 2.2, a natural soil. When assessing the toxic effects in the environment, soil properties are often different from those in standard soils, which might lead to different exposure situation for the test species and therefore to misleading conclusions. Selected to cover a broad range of properties and based on the Euro-Soils concept, 17 different soils were studied regarding their suitability to two tes species: Folsomia candida and Hypogastrura assimilis (Collembola). In reproduction tests, the test species reacted differently to the soils. F. candida was less affected by soil properties: 500 to 1200 juveniles/vessel were found in untreated soils (i.e. controls). These differences can be attributed to normal interindividual variability. H. assimilis showed a significant correlation with maximum water-holding capacity and also a tendency to lower the reproductive output in soils with a low pH (<4). Therefore, some soils were revealed to be inappropriate for tests with H. assimilis. In the main tests, the effect of the reference test substance Phenmedipham (formulation Betosyp) was studied in those soils where sufficient reproduction was determined beforehand. Clearly, the chronic end point was more sensitive than survival when testing Phenmedipham. In H. assimilis, because of high variability and low effects of tested dosages, no conclusions could be drawn. In F. candida, different soils caused different toxic effects: Juveniles preferred soils with high C-to-N ratios. Higher microbial activity might support a quicker metabolization of the test substance. In general, the toxic response is caused by a synergistic action of several soil properties with each of them exerting an effect too small to be clarified with the available set of data.