The veterinary parasiticide ivermectin was selected as a case study compound within the project ERAPharm (Environmental Risk Assessment of Pharmaceuticals). Based on experimental data generated within ERAPharm and additional literature data, an environmental risk assessment (ERA) was performed mainly according to international and European guidelines. For the environmental compartments surface water, sediment, and dung, a risk was indicated at all levels of the tiered assessment approach. Only for soil was no risk indicated after the lower tier assessment. However, the use of effects data from additional 2-species and multispecies studies resulted in a risk indication for collembolans. Although previously performed ERAs for ivermectin revealed no concern for the aquatic compartment, and transient effects on dung-insect populations were not considered as relevant, the present ERA clearly demonstrates unacceptable risks for all investigated environmental compartments and hence suggests the necessity of reassessing ivermectin-containing products. Based on this case study, several gaps in the existing guidelines for ERA of pharmaceuticals were shown and improvements have been suggested. The action limit at the start of the ERA, for example, is not protective for substances such as ivermectin when used on intensively reared animals. Furthermore, initial predicted environmental concentrations (PECs) of ivermectin in soil were estimated to be lower than refined PECs, indicating that the currently used tiered approach for exposure assessment is not appropriate for substances with potential for accumulation in soil. In addition, guidance is lacking for the assessment of effects at higher tiers of the ERA, e.g., for field studies or a tiered effects assessment in the dung compartment.
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.
Clusia hilariana Schlechtendal is a dioecious tree performing crassulacean acid metabolism (CAM). It grows in the sandy coastal restinga of Brazil. As pioneers and 'nurse plants' individual trees become surrounded by other vegetation forming isolated vegetation islands on the sand plain. The female plants produce large and heavy fleshy fruits suggesting a particular demand for resources. Frugivores may introduce nutrients (faeces) and diaspores into the stands of female plants. Thus, the hypothesis was advanced that sex expression in C. hilariana has important general consequences for the physiology of the trees themselves and also determines aspects of the vegetation around them. Field work in the restinga of Macaé (Rio de Janeiro State, Brazil) showed that morphological leaf characteristics, the expression of CAM and the species diversity and species composition of their respective vegetation islands were similar for both sexes. The female plants had somewhat higher chlorophyll levels. Differences in effective quantum yield of photosystem II (PS II) and non-photochemical quenching of chlorophyll a fluorescence between female and male plants were very small. Female plants were a little less subject to acute photoinhibition at noon. Maximum fluorescence (Fm), variable fluorescence (Fv) and minimum fluorescence (F 0 ) were significantly lower in female plants. However, maximum quantum yield ratio of male to female plants as well as fluorescence yield ratio and minimum fluorescence ratio of male to female plants related to chlorophyll content were all close to unity. Therefore, although there may be some fine-tuned differences in the light reactions of photosynthesis, PS II of male and female plants are not different. Hence, the hypothesis of possible important consequences of sex determination for expression of morphological and physiological traits of the plants and for the composition of the 'nursed' vegetation underneath them must be rejected.
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