Abstract: The aim of the present paper was to examine in field conditions the feasibility of the experimentally developed behavioural tests for assessment of two temperamental traits of cold blood horses: fearfulness and reactivity to humans. The predictive, convergent and discriminant validity of the tests was evaluated. Fifty-three Polish Cold Blood Horses (CBHs) were tested with the battery of behavioural tests involving passive and active contact with humans, handling, startling, novel object and surface tests. The behaviour of horses in real, on-farm conditions was evaluated with the caretaker's questionnaire (CQ). The tests demonstrated that a non-startling object and unforced human manipulations were willingly accepted by horses. In contrast, the horses were more reluctant to interact spontaneously with humans and showed longer persistence of a withdrawal reaction when presented with a startling stimulus or when restrained. CQ confirmed the common opinion of calmness, patience and good manageability of the CBHs. The results of behavioural tests were correlated with the scores derived from factor analysis of the CQ. Moreover, the majority of tests measures were correlated within reactivity to humans (median rs = 0.56) and fear tests (median rs = 0.41). Thus, predictive and convergent validity of the employed tests has been confirmed. However, reactivity to humans and fear measures were also correlated (median rs = 0.36), therefore discriminant validity of the tests used has been not confirmed. Therefore, we suggest that the parameters of tests designed to measure two separate traits: reactivity to humans and fearfulness, measured in fact avoidance of potential harm and withdrawal behaviour. Although selection for the work-appropriate behaviour is almost nonexistent nowadays, we conclude that Polish Cold Blood Horses are still predisposed for disciplines of equitation and driving, which demand an easy to handle horse. Nonetheless, we recommend behavioural testing of horses when approving for breeding. For this purpose, we can confirm usefulness of the experimentally developed tests in field assessment of general behavioural reactivity of horses.

Abstract: Horses’ fear reactions are hazardous to both horses and human beings, but it is not clear whether fear is influenced more by training or by other factors such as genetics. The following study was designed to detect differences between young, untrained (U) and older, well-trained (T) horses of dressage (D), show-jumping (J), and mixed (M) genetic lines with regard to intensity of reaction and ease of habituation to a frightening stimulus. In five consecutive trials, 90 horses were exposed to a standardized fear-eliciting stimulus where intensity and duration of the reactions were recorded. Repeated measures analysis showed that flight reactions by J were less intense (p < 0.05) than those by D or M regardless of training status or age. Habituation to the stimulus over time was not significantly (p > 0.1) different between the disciplines, as indicated by similar slopes for all measurements, but reaction vigour declined faster for T than for U. These findings indicate that there may be a genetic basis for less strong, though not shorter-lasting, fear reactions in J compared to D or M lines of horses. Research including the estimation of genetic correlations between traits related to fearfulness and to performance would be required to verify this assumption.

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.

Abstract: All feces produced during 24 h were collected from five pigs and cortisol and immunoreactive cortisol metabolites (CICM), and IgA were quantified. Within pigs, the concentrations of CICM and IgA varied extensively between random samples obtained from a single fecal dropping, and deviated in most cases significantly from the true concentration measured in total fecal output (CV 6.7–130%). The CICM and IgA contents varied considerably (CV 8.1–114%) within and between individual fecal droppings from the same pig compared to the total fecal excretion. In conclusion, single random samples could not be used to reliably quantify the total fecal concentration or excretion of CICM or IgA in pigs. Analyses of all feces collected during shorter periods than 24 h did not provide an accurate estimate of the daily excretion of CICM. Thus, the concentration of stress sensitive molecules in random single fecal samples as an indicator of animal welfare should be interpreted with prudence.

Abstract: Abstract The right brain hemisphere, connected to the left eye, coordinates fight and flight behaviors in a wide variety of vertebrate species. We investigated whether left eye vision predominates in dairy cows’ interactions with other cows and humans, and whether dominance status affects the extent of visual lateralization. Although we found no overall lateralization of eye use to view other cows during interactions, cows that were submissive in an interaction were more likely to use their left eye to view a dominant animal. Both subordinate and older cows were more likely to use their left eye to view other cattle during interactions. Cows that predominantly used their left eye during aggressive interactions were more likely to use their left eye to view a person in unfamiliar clothing in the middle of a track by passing them on the right side. However, a person in familiar clothing was viewed predominantly with the right eye when they passed mainly on the left side. Cows predominantly using their left eyes in cow-to-cow interactions showed more overt responses to restraint in a crush compared with cows who predominantly used their right eyes during interactions (crush scores: left eye users 7.9, right eye users 6.4, standard error of the difference = 0.72). Thus, interactions between 2 cows and between cows and people were visually lateralized, with losing and subordinate cows being more likely to use their left eyes to view winning and dominant cattle and unfamiliar humans.