Registration of veterinary medical products includes the provision that field tests may be required to assess potential nontarget effects associated with the excretion of product residues in dung of treated livestock (phase II, tier B testing). However, regulatory agencies provide no guidance on the format of these tests. In the present study, the authors report on the development of a standardized field test method designed to serve as a tier B test. Dung was collected from cattle before and up to 2 mo after treatment with a topical application of a test compound (ivermectin). Pats formed of dung from the different treatments were placed concurrently in the field to be colonized by insects. The abundance, richness, and diversity of insects developing from egg to adult in these pats were compared across treatments using analysis of variance tests. Regression analyses were used to regress abundance, richness, and diversity against residue concentrations in each treatment. Results of the regression were used to estimate mean lethal concentration (LC50) values. The robustness of the method and the repeatability of its findings were assessed concurrently in 4 countries (Canada, France, Switzerland, and The Netherlands) in climatically diverse ecoregions. Results were generally consistent across countries, and support the method's formal adoption by the European Union to assess the effects of veterinary medical product residues on the composition and diversity of insects in dung of treated livestock. Environ Toxicol Chem 2016;35:1934-1946. © 2015 Crown in the right of Canada. Published by Wiley Periodicals Inc., on behalf of SETAC.

Cattle treated with the veterinary parasiticide ivermectin fecally excrete residues. The authors report the exposition and dissipation characteristics of these residues in dung of ivermectin-treated cattle and in soil beneath this dung on pastures in Canada, France, Switzerland, and The Netherlands. Residues were quantified for dung collected from cattle after 3 d, 7 d, 14 d, and 28 d posttreatment and subsequently exposed in the field for up to 13 mo. The authors optimized a high-performance liquid chromatography-fluorescence detection method to detect ivermectin residues in dung and soil matrices. They showed that a solid phase extraction and purification step generally can be eliminated to reduce the time and cost of these analyses. They also found that the addition of water to relatively dry samples improves the extraction efficiency of residues. They then analyzed the field samples to document differences in ivermectin dissipation in cattle dung among sites, with 50% dissipation times of up to 32 d and 90% dissipation times >396 d. They further showed that the dissipation characteristics of residues are comparable between dung of ivermectin-treated cattle and dung to which ivermectin has been added directly. Lastly, they report the first use of a desorption electrospray ionization-high-resolution-mass spectrometric method to detect residues of metabolites in a dung matrix. Environ Toxicol Chem 2016;35:1924-1933. © 2016 SETAC.

By degrading the dung of livestock that graze on pastures, coprophilous arthropods accelerate the cycling of nutrients to maintain pasture quality. Many veterinary medicinal products, such as ivermectin, are excreted unchanged in the dung of treated livestock. These residues can be insecticidal and may reduce the function (i.e., dung-degradation) of the coprophilous community. In the present study, we used a standard method to monitor the degradation of dung from cattle treated with ivermectin. The present study was performed during a 1-yr period on pastures in Canada, France, The Netherlands, and Switzerland. Large effects of residue were detected on the coprophilous community, but degradation of dung was not significantly hampered. The results emphasize that failure to detect an effect of veterinary medicinal product residues on dung-degradation does not mean that the residues do not affect the coprophilous community. Rather, insect activity is only one of many factors that affect degradation, and these other factors may mask the nontarget effect of residues. Environ Toxicol Chem 2016;35:1953-1958. © 2015 SETAC.

The authorization of veterinary medicinal products requires that they be assessed for nontarget effects in the environment. Numerous field studies have assessed these effects on dung organisms. However, few studies have examined effects on soil-dwelling organisms, which might be exposed to veterinary medicinal product residues released during dung degradation. The authors compared the abundance of earthworms and springtails in soil beneath dung from untreated cattle and from cattle treated 0 d, 3 d, 7 d, 14 d, and 28 d previously with ivermectin. Study sites were located in different ecoregions in Switzerland (Continental), The Netherlands (Atlantic), France (Mediterranean), and Canada (Northern Mixed Grassland). Samples were collected using standard methods from 1 mo to 12 mo after pat deposition. Ivermectin concentrations in soil beneath dung pats ranged from 0.02 mg/kg dry weight (3 mo) to typically <0.006 mg/kg dry weight (5-7 mo). Earthworms were abundant and species-rich at the Swiss and Dutch sites, less common with fewer species at the French site, and essentially absent at the Canadian site. Diverse but highly variable communities of springtails were present at all sites. Overall, results showed little effect of residues on either earthworms or springtails. The authors recommend that inclusion of soil organisms in field studies to assess the nontarget effects of veterinary medicinal products be required only if earthworms or springtails exhibit sensitivity to the product in laboratory tests. Environ Toxicol Chem 2016;35:1959-1969. © 2015 SETAC.

Properly designed (randomized and/or balanced) experiments are standard in ecological research. Molecular methods are increasingly used in ecology, but studies generally do not report the detailed design of sample processing in the laboratory. This may strongly influence the interpretability of results if the laboratory procedures do not account for the confounding effects of unexpected laboratory events. We demonstrate this with a simple experiment where unexpected differences in laboratory processing of samples would have biased results if randomization in DNA extraction and PCR steps do not provide safeguards. We emphasize the need for proper experimental design and reporting of the laboratory phase of molecular ecology research to ensure the reliability and interpretability of results.