An experimental test of the habitat-amount hypothesis for saproxylic beetles in a forested region.

The habitat-amount hypothesis challenges traditional concepts that explain species richness within habitats, such as the habitat-patch hypothesis, where species number is a function of patch size and patch isolation. It posits that effects of patch size and patch isolation are driven by effects of sample area, and thus that the number of species at a site is basically a function of the total habitat amount surrounding this site. We tested the habitat-amount hypothesis for saproxylic beetles and their habitat of dead wood by using an experiment comprising 190 plots with manipulated patch sizes situated in a forested region with a high variation in habitat amount (i.e., density of dead trees in the surrounding landscape). Although dead wood is a spatio-temporally dynamic habitat, saproxylic insects have life cycles shorter than the time needed for habitat turnover and they closely track their resource. Patch size was manipulated by adding various amounts of downed dead wood to the plots (~800 m³ in total); dead trees in the surrounding landscape (~240 km2 ) were identified using airborne laser scanning (light detection and ranging). Over 3 yr, 477 saproxylic species (101,416 individuals) were recorded. Considering 20-1,000 m radii around the patches, local landscapes were identified as having a radius of 40-120 m. Both patch size and habitat amount in the local landscapes independently affected species numbers without a significant interaction effect, hence refuting the island effect. Species accumulation curves relative to cumulative patch size were not consistent with either the habitat-patch hypothesis or the habitat-amount hypothesis: several small dead-wood patches held more species than a single large patch with an amount of dead wood equal to the sum of that of the small patches. Our results indicate that conservation of saproxylic beetles in forested regions should primarily focus on increasing the overall amount of dead wood without considering its spatial arrangement. This means dead wood should be added wherever possible including in local landscapes with low or high dead-wood amounts. For species that have disappeared from most forests owing to anthropogenic habitat degradation, this should, however, be complemented by specific conservation measures pursued within their extant distributional ranges.

Results of extended plant tests using more realistic exposure scenarios for improving environmental risk assessment of veterinary pharmaceuticals.

BACKGROUND: Residues of veterinary medicinal products (VMPs) enter the environment via application of manure onto agricultural areas where in particular antibiotics can cause phytotoxicity. Terrestrial plant tests according to OECD guideline 208 are part of the environmental risk assessment of VMPs. However, this standard approach might not be appropriate for VMPs which form non-extractable residues or transformation products in manure and manure-amended soil. Therefore, a new test design with a more realistic exposure scenario via manure application is needed. This paper presents an extended plant test and its experimental verification with the veterinary antibiotics florfenicol and tylosin tartrate. With each substance, plant tests with four different types of application were conducted: standard tests according to OECD 208 and three tests with application of test substance via spiked manure either without storage, aerobically incubated, or anaerobically incubated for different time periods. RESULTS: In standard tests, the lowest NOEC was <0.06 mg/kg dry soil for florfenicol and 16.0 mg/kg dry soil for tylosin tartrate. Pre-tests showed that plant growth was not impaired at 22-g fresh manure/kg dry soil, which therefore was used for the final tests. The application of the test substances via freshly spiked as well as via aerobically incubated manure had no significant influence on the test results. Application of florfenicol via anaerobically incubated manure increased the EC10 by a factor up to 282 and 540 for half-maximum and for maximum incubation period, respectively. For tylosin tartrate, this factor amounted to 64 at half-maximum and 61 at maximum incubation period. The reduction of phytotoxicity was generally stronger when using cattle manure than pig manure and particularly in tests with cattle manure phytotoxicity decreased over the incubation period. CONCLUSIONS: The verification of the extended plant test showed that seedling emergence and growth are comparable to a standard OECD 208 test and reliable effect concentrations could be established. As demonstrated in the present study, phytotoxicity of veterinary antibiotics can be significantly reduced by application via incubated manure compared to the standard plant test. Overall, the presented test design proved suitable for inclusion into the plant test strategy for VMPs.

Fate and effects of ivermectin on soil invertebrates in terrestrial model ecosystems.

The effect of ivermectin on soil organisms was assessed in Terrestrial Model Ecosystems (TMEs). Intact soil cores were extracted from a pasture in England and kept for up to 14 weeks in the laboratory. Ivermectin was applied to the soil surface via spiked cow dung slurry at seven concentration rates ranging from 0.25 to 180 mg/TME, referring to concentrations of 0.19-227 mg ivermectin/kg soil dry weight in the uppermost (0-1 cm) soil layer. After 7, 28 and 96 days following the application soil cores were destructively sampled to determine ivermectin residues in soil and to assess possible effects on microbial biomass, nematodes, enchytraeids, earthworms, micro-arthropods, and bait-lamina feeding activity. No significant effect of ivermectin was found for microbial respiration and numbers of nematodes and mites. Due to a lack of dose-response patterns no effect concentrations could be determined for the endpoints enchytraeid and collembolan numbers as well as total earthworm biomass. In contrast, EC50 values for the endpoint feeding rate could be calculated as 0.46, 4.31 and 15.1 mg ivermectin/kg soil dry weight in three soil layers (0-1, 0-5 and 0-8 cm, respectively). The multivariate Principal Response Curve (PRC) was used to calculate the NOEC(community), based on earthworm, enchytraeid and collembolan abundance data, as 0.33 and 0.78 mg ivermectin/kg soil dw for day 7 and day 96, respectively. The results shown here are in line with laboratory data, indicating in general low to moderate effects of ivermectin on soil organisms. As shown by the results of the bait-lamina tests, semi-field methods such as TMEs are useful extensions of the battery of potential test methods since complex and ecologically relevant endpoints can be included.

Ring-testing and field-validation of a terrestrial model ecosystem (TME)--an instrument for testing potentially harmful substances: effects of carbendazim on soil microbial parameters.

The effects of carbendazim on substrate induced respiration (SIR), dehydrogenase activity (DHA), phosphatase activity and thymidine incorporation by bacteria were evaluated in an experiment with an open intact Terrestrial Model Ecosystem (TME) and in a simultaneous field-validation study. Experiments were performed on four different European soils in Germany, The Netherlands, United Kingdom and Portugal. Data analysis focused on (i) detecting differences between experiments, especially in control values, (ii) checking similarity in data variability at each treatment level between experiments and (iii) analysing the resemblance of response to the model chemical in both experiments. Results obtained showed that control values from TME experiments were similar to those obtained on the respective field site, in most of the comparisons made for SIR, DHA and thymidine incorporation. Phosphatase activity revealed more differences, but values of both experiments had the same order of magnitude. At least part of the variation could be explained from the correlation of the microbial parameters with soil moisture content. Comparisons on data variability also revealed the absence of significant differences between experiments in all parameters in most cases, indicating that TMEs were able to represent the spatial variability found in the field. Effects of carbendazim, when occurring, were observed at treatment levels exceeding the highest recommended application rate of 0.36 kg a.i./ha. Effects on SIR and DHA were observed early in time, but effects on phosphatase activity and thymidine incorporation rate were found 8 or 16 weeks after chemical application. These effects were mild, and rarely a 50% inhibition on any of these parameters was seen at carbendazim dosages up to 87.5 kg a.i./ha. The response to the model chemical in TMEs and field plots was similar in most cases. These results give promising prospects for the use of TMEs as an integrative tool in higher tier levels of different assessment schemes.

Ring-testing and field-validation of a terrestrial model ecosystem (TME)--an instrument for testing potentially harmful substances: fate of the model chemical carbendazim.

The fate of the fungicide carbendazim (applied in the formulation Derosal) in soil was determined in Terrestrial Model Ecosystem (TME) tests and corresponding field-validation studies, which were performed in four different countries (United Kingdom, Germany, Portugal, and The Netherlands). The tests used different soil types, and lasted for 16 weeks. On three of the four sites, grassland soils were used while the fourth site had an arable soil. TMEs consisted of intact soil columns (diameter 17.5 cm; length 40 cm) and were taken from the site where the field study was performed. In the first series of TME tests, carbendazim was applied at four dosages ranging between 0.36 and 77.8 kg a.i./ha, while in the second series of TME tests and the field-validation studies six dosages between 0.36 and 87.5 kg a.i./ha were applied. DT50 values for the dissipation of carbendazim in the TME and field tests were in most cases not significantly affected by the dosage used and ranged between 3.1 and 13.9 weeks in the top 15 cm soil layers. Corresponding DT90 values ranged between 10.1 and 46.1 weeks. DT50 and DT90 values tended to be higher in the more acidic soils of Amsterdam and Flörsheim (pH-KCl 4.8-5.1 and 5.3-5.9, respectively) than in the less acidic soils of Bangor and Coimbra (pH-KCl 5.8-6.6 and 6.4-7.1, respectively). Fate of carbendazim in soil showed similar patterns in the two TME tests and the corresponding field-validation study performed at each site. The only exception was Flörsheim, where the compound was significantly more persistent in the field probably due to different climatic conditions. Carbendazim was not recovered from leachates produced in the TME tests, nor was the compound detected in soil layers deeper than 15 cm. This demonstrates that no significant leaching occurred. This study demonstrates the the TME tests were quite successful in predicting the fate of carbendazim under field conditions.

Ring-testing and field-validation of a terrestrial model ecosystem (TME)--an instrument for testing potentially harmful substances: effects of carbendazim on organic matter breakdown and soil fauna feeding activity.

Organic matter (OM) decomposition and soil fauna feeding activity were integrated as functional endpoints into ecotoxicological tests with intact-soil-core Terrestrial Model Ecosystems (TMEs). Cellulose filter paper served as standardized OM and was either inserted into the top soil or placed on the soil surface for a period of up to 16 weeks. Faunal feeding activity was assessed by the bait-lamina method. The fungicide carbendazim, applied at six dosages ranging from 0.36 kg/ha to 87.5 kg a.i./ha, served as a model chemical. To validate the results from the TME test, a field study was run in parallel. In TMEs the cellulose paper inserted into the soil was decomposed faster than under field conditions. The carbendazim-induced effects on OM decomposition in TMEs and in the field were comparable and followed a clear dose-response relationship. The calculated EC50 values after 8 weeks of incubation were 9.5, 7.1 and 2.1 kg carbendazim/ha for grassland TMEs, grassland field and arable TMEs, respectively. The feeding activity of the soil fauna showed a large variability. The EC50 values for the effect of carbendazim on bait-lamina consumption ranged between 2.0 and 56 kg a.i./ha. Effects on decomposition were correlated with effects on enchytraeids and earthworms but not with effects on bait-lamina consumption.

Ring-testing and field-validation of a terrestrial model ecosystem (TME)--an instrument for testing potentially harmful substances: conceptual approach and study design.

During spring and summer 1999 a ring-test and field-validation study with an open, intact Terrestrial Model Ecosystem (TME) was conducted at four different European sites (Amsterdam, The Netherlands; Bangor, U.K.; Coimbra, Portugal; Flörsheim, Germany). The objective of the study was to establish a standardised method which allows the impact of chemical stressors on terrestrial compartments at ecosystem level to be investigated and possible uses of such data in existing Environmental Risk Assessments (ERAs) for chemicals to be evaluated. This issue of Ecotoxicology presents in a series of papers the results of the TME ring-test and field-validation study. Additionally, results derived from an open-homogeneous terrestrial microcosm (Integrated Soil Microcosm, ISM) are included in this series as a separate paper. In this first paper of the series background information on the planning and organisation of the study are given. The conceptual approach and the design of the study with TMEs are briefly outlined, based on the scientific discussion on the use of terrestrial microcosms in ecology and applied environmental sciences during the last 25 years. Further, some suggestions are presented on the selection of measurement endpoints to quantify structural and functional aspects of terrestrial ecosystems. Finally, the main results of the TME-study are summarised and conclusions are drawn on the technical feasibility of TMEs, their comparability with field studies and the potential use of TMEs in ERA.