Abstract: Context. Livestock producers worldwide are negatively affected by livestock losses because of predators and wildlife-transmitted diseases. In the western Great Lakes Region of the United States, this conflict has increased as grey wolf (Canis lupus) populations have recovered and white-tailed deer (Odocoileus virginianus) have served as a wildlife reservoir for bovine tuberculosis (Myobacterium bovis).Aims. We conducted field experiments on cattle farms to evaluate the effectiveness of livestock-protection dogs (LPDs) for excluding wolves, coyotes (C. latrans), white-tailed deer and mesopredators from livestock pastures.Methods. We integrated LPDs on six cattle farms (treatment) and monitored wildlife use with tracking swaths on these farms, concurrent with three control cattle farms during 2005-2008. The amount of time deer spent in livestock pastures was recorded using direct observation.Key results. Livestock pastures protected by LPDs had reduced use by these wildlife compared with control pastures not protected by LPDs. White-tailed deer spent less time in livestock pastures protected by LPDs compared with control pastures not protected by LPDs.Conclusions. Our research supports the theory that LPDs can be an effective management tool for reducing predation and disease transmission. We also demonstrate that LPDs are not limited to being used only with sheep and goats; they can also be used to protect cattle.Implications. On the basis of our findings, we support the use of LPDs as a proactive management tool that producers can implement to minimise the threat of livestock depredations and transmission of disease from wildlife to livestock. LPDs should be investigated further as a more general conservation tool for protecting valuable wildlife, such as ground-nesting birds, that use livestock pastures and are affected by predators that use these pastures.

Abstract: Faecal steroid hormone metabolites are becoming increasingly popular as parameters for reproductive functions and stress. The extraction of the steroids from the faecal matrix represents the initial step before quantification can be performed. The steroid metabolites present in the faecal matrix are of varying polarity and composition, so selection of a proper extraction procedure is essential. There have been some studies to address this complex but often neglected point. Radiolabelled steroids (e.g. cortisol or progesterone) have frequently been added to faecal samples to estimate the efficiency of the extraction procedures used. However, native, unmetabolized steroids are normally not present in the faeces and therefore the results are artifi- cial and do not accurately reflect the actual recoveries of the substances of interest. In this respect, recovery experiments based on faecal samples from radiometabolism studies are more informative. In these samples, the metabolite content accurately reflects the mixture of metabolites present in the given species. As a result, it is possible to evaluate different extraction methods for use with faecal samples. We present studies on sheep, horses, pigs, hares and dogs that utilized samples containing naturally metabolized, 14C-labelled steroids. We recommend extracting faecal steroids by simply suspending the faeces in a high percentage of a primary alcohol (for glucocorticoid metabolites 80% aqueous methanol proved best suited for virtually all mammalian species tested so far). Not only does the procedure significantly increase the total amount of recovered radioactivity, it also increases the percentage of unconjugated metabolites, which are more likely to be recognized by the antibodies used in various immunoassays. The advantages of this extraction procedure are clear: it is very easy to use (no evaporation step is needed), it yields high recoveries and variation based on the extraction procedure is reduced to a minimum.

Abstract: Faecal steroid hormone metabolites are becoming increasingly popular as parameters for reproductive functions and stress. Theextraction of the steroids from the faecal matrix represents the initial step before quantification can be performed. The steroid metabolites present in the faecal matrix are of varying polarity and composition, so selection of a proper extraction procedure is essential. There have been some studies to address this complex but often neglected point. Radiolabelled
steroids (e.g. cortisol or progesterone) have frequently been added to faecal samples to estimate the efficiency of the extraction procedures used. However, native, unmetabolized steroids are normally not present in the faeces and therefore the results are artificial and do not accurately reflect the actual recoveries of the substances of interest. In this respect, recovery experiments based on faecal samples from radiometabolism studies are more informative. In these samples, the metabolite content accurately reflects the mixture of metabolites present in the given species. As a result, it is possible to evaluate different extraction methods for use with faecal samples. We present studies on sheep, horses, pigs, hares and dogs that utilized samples containing naturally metabolized, 14C-labelled steroids.

Abstract: Abstract Only few quantitative reports exist about the concentrations and induction of immunoglobulin A (IgA) in mucosal secretions of horses. Despite this, it is widely assumed that IgA is the predominant immunoglobulin on mucosal surfaces in the horse. Here, two new monoclonal antibodies (mAbs) against equine IgA, clones 84-1 and 161-1, were developed and characterized in detail. Both IgA mAbs specifically bound monomeric and dimeric equine IgA in different applications, such as Western blots and fluorescent bead-based assays. Cross-reactivity with other equine immunoglobulin isotypes was not observed. The new IgA mAb 84-1 was used in combination with the previously characterized anti-equine IgA mAb BVS2 for the development and validation of a fluorescent bead-based assay to quantify total IgA in equine serum and various secretions. The IgA assay's linear detection ranged from 64 pg/ml to 1000 ng/ml. For the quantification of IgA in serum or in secretions an IgA standard was purified from serum or nasal wash fluid (secretory IgA), respectively. The different standards were needed for accurate IgA quantification in the respective samples taking the different signal intensities of monomeric and dimeric IgA on the florescent bead-based assay into account. IgA was quantified by the bead-based assay established here in different equine samples of healthy adult individuals. In serum the median total IgA was 0.45 mg/ml for Thoroughbred horses (TB, n = 10) and 1.16 mg/ml in Icelandic horses (ICH, n = 12). In nasopharyngeal secretions of TB (n = 7) 0.13 mg/ml median total IgA was measured, and 0.25 mg/ml for ICH (n = 12). Saliva of ICH (n = 6) contained a median of 0.15 mg/ml, colostrum of Warmbloods (n = 8) a median of 1.89 mg/ml IgA. Compared to IgG1 and IgG4/7 quantified in the same samples, IgA appeared as the major immunoglobulin isotype in nasopharyngeal secretions and saliva while it is a minor isotype in serum and colostrum. The newly developed monoclonal antibodies against equine IgA and the resulting bead-based assay for quantification of total IgA can notably improve the evaluation of mucosal immunity in horses.