Abstract: The conditions under which horses are kept and the performance of acts in the circus ring may give rise to animal protection-relevant aspects for circus and show horses. A number of intolerable conditions under which horses are kept and procedures adopted for the work with circus and show horses are described. In addition, attention is drawn to monitoring methods capable of exposing the deplorable shortcomings of these businesses.

Abstract: Adult male elephants periodically show the phenomenon of musth, a condition associated with increased aggressiveness, restlessness, significant weight reduction and markedly elevated androgen levels. It has been suggested that musth-related behaviours are costly and that therefore musth may represent a form of physiological stress. In order to provide data on this largely unanswered question, the first aim of this study was to evaluate different assays for non-invasive assessment of adrenocortical function in the male African elephant by (i) characterizing the metabolism and excretion of [3H]cortisol (3H-C) and [14C]testosterone (14C-T) and (ii) using this information to evaluate the specificity of four antibodies for determination of excreted cortisol metabolites, particularly with respect to possible cross-reactions with androgen metabolites, and to assess their biological validity using an ACTH challenge test. Based on the methodology established, the second objective was to provide data on fecal cortisol metabolite concentrations in bulls during the musth and non-musth condition. 3H-C (1 mCi) and 14C-T (100 microCi) were injected simultaneously into a 16 year old male and all urine and feces collected for 30 and 86 h, respectively. The majority (82%) of cortisol metabolites was excreted into the urine, whereas testosterone metabolites were mainly (57%) excreted into the feces. Almost all radioactive metabolites recovered from urine were conjugated (86% 3H-C and 97% 14C-T). In contrast, 86% and >99% of the 3H-C and 14C-T metabolites recovered from feces consisted of unconjugated forms. HPLC separations indicated the presence of various metabolites of cortisol in both urine and feces, with cortisol being abundant in hydrolysed urine, but virtually absent in feces. Although all antibodies measured substantial amounts of immunoreactivity after HPLC separation of peak radioactive samples and detected an increase in glucocorticoid output following the ACTH challenge, only two (in feces against 3alpha,11-oxo-cortisol metabolites, measured by an 11-oxo-etiocholanolone-EIA and in urine against cortisol, measured by a cortisol-EIA) did not show substantial cross-reactivity with excreted 14C-T metabolites and could provide an acceptable degree of specificity for reliable assessment of glucocorticoid output from urine and feces. Based on these findings, concentrations of immunoreactive 3alpha,11-oxo-cortisol metabolites were determined in weekly fecal samples collected from four adult bulls over periods of 11-20 months to examine whether musth is associated with increased adrenal activity. Results showed that in each male levels of these cortisol metabolites were not elevated during periods of musth, suggesting that in the African elephant musth is generally not associated with marked elevations in glucocorticoid output. Given the complex nature of musth and the variety of factors that are likely to influence its manifestation, it is clear, however, that further studies, particularly on free-ranging animals, are needed before a possible relationship between musth and adrenal function can be resolved. This study also clearly illustrates the potential problems associated with cross-reacting metabolites of gonadal steroids in EIAs measuring glucocorticoid metabolites. This has to be taken into account when selecting assays and interpreting results of glucocorticoid metabolite analysis, not only for studies in the elephant but also in other species.

Abstract: OBJECTIVE: To determine whether therapeutic riding resulted in higher levels of stress or frustration for horses than did recreational riding and whether therapeutic riding with at-risk individuals was more stressful for the horses than was therapeutic riding with individuals with physical or emotional handicaps. DESIGN: Observational study. ANIMALS: 14 horses in a therapeutic riding program. PROCEDURE: An ethogram of equine behaviors was created, and horses were observed while ridden by 5 groups of riders (recreational riders, physically handicapped riders, psychologically handicapped riders, at risk children, and special education children). Number of stress-related behaviors (ears pinned back, head raised, head turned, head tossed, head shaken, head down, and defecation) was compared among groups. RESULTS: No significant differences in mean number of stress-related behaviors were found when horses were ridden by recreational riders, physically handicapped riders, psychologically handicapped riders, or special education children. However, mean number of stress-related behaviors was significantly higher when horses were ridden by the at-risk children. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that for horses in a therapeutic riding program, being ridden by physically or psychologically handicapped individuals is no more stressful for the horses than is being ridden in the same setting by recreational riders. However, at-risk children caused more stress to the horses, suggesting that the time horses are ridden by at-risk children should be limited both daily and weekly.

Abstract: The partition of human antibodies in aqueous two-phase systems (ATPSs) of polyethylene glycol (PEG) and phosphate was systematically studied using first pure proteins systems and then an artificial mixture of proteins containing 1mg/ml human immunoglobulin G (IgG), 10mg/ml serum albumin and 2mg/ml myoglobin. Preliminary results obtained using pure proteins systems indicated that the PEG molecular weight and concentration, the pH value and the salts concentration had a pronounced effect on the partitioning behaviour of all proteins. For high ionic strengths and pH values higher than the isoelectric point (pI) of the contaminant proteins, IgG could be selectively recovered on the top phase. According to these results, a face centred composite design was performed in order to optimise the purification of IgG from the mixture of proteins. The optimal conditions for the isolation of IgG were observed for high concentrations of NaCl and low concentrations of both phase forming components. The best purification was achieved using an ATPS containing 8% (w/w) PEG 3350, 10% (w/w) phosphate pH 6 and 15% (w/w) NaCl. A recovery yield of 101+/-7%, a purity of 99+/-0% and a yield of native IgG of 97+/-4% were obtained. Back extraction studies of IgG to a new phosphate phase were performed and higher yields were obtained using 10% phosphate buffer at pH 6. The total extraction yield was 76% and the purity 100%.

Abstract: A wireless acceleration measurement system was applied to free-moving cows and horses. Sensors were available as a collar and a flat box for measuring leg or trunk movements. Results were transmitted simultaneously by radio or stored in an 8-MB internal memory. As analytical procedures, frequency distributions with standard deviations, spectral analyses, and fractal analyses were applied. Bymeans of the collar sensor, basic behavior patterns (standing, grazing, walking, ruminating, drinking, and hay uptake) could be identified in cows. Lameness could be detected in cows and horses by means of the leg sensor. The portion of basic and harmonic spectral components was reduced; the fractal dimension was reduced. The system can be used for the detection and analysis of even small movements of free-moving humans or animals over several hours. It is convenient for the analysis of basic behaviors, emotional reactions, or events causing flight or fright or for comparing different housing elements, such as floors or fences.