Abstract: This study aimed to increase the knowledge on salivary antibodies in the horse since these constitute an important part of the immune defence of the oral cavity. For that purpose assays to detect horse immunoglobulin A (IgA) including secretory IgA (SIgA) were set up and the molecular weights of different components of the horse IgA system were estimated. Moreover, samples from 51 clinically healthy horses were tested for total SIgA and IgG amounts in saliva and relative IgG3/5 (IgG(T)) and IgG4/7 (IgGb) content were tested in serum and saliva. Results showed a mean concentration of 74μg SIgA/ml horse saliva and that there was a large inter-individual variation in salivary SIgA concentration. For total IgG the mean concentration was approx. 5 times lower than that of SIgA, i.e. 20μg IgG/ml saliva and the inter-individual variation was lower than that observed for SIgA. The saliva-serum ratio for IgG isotypes IgG3/5 and IgG4/7 was also assessed in the sampled horses and this analysis showed that the saliva-serum ratio of IgG4/7 was in general approximately 4 times higher than that of IgG3/5. The large inter-individual variation in salivary SIgA levels observed for the normal healthy horses in the present study emphasises the need for a large number of observations when studying this parameter especially in a clinical setting. Moreover, our results also indicated that some of the salivary IgG does not originate from serum but may be produced locally. Thus, these results provide novel insight, and a base for further research, into salivary antibody responses of horses.

Abstract: Glucocorticoids (GCs; i.e. cortisol/corticosterone) are a central component of the stress response and thus their measurement is frequently used to evaluate the impact of stressful situations. Their metabolites from faeces of various animal species are more and more taken as a non-invasive aid to assess GC release and thus adrenocortical activity. The current literature review includes an extensive collection (1327 papers) and evaluation (see also Supplementary Tables) of the literature on faecal cortisol/corticosterone metabolite (FCM) analysis published to date. It aims at giving reference for researchers interested in implementing FCM analysis into their study or seeking to improve such methods by providing background knowledge on GC metabolism and excretion, conveying insights into methodological issues and stating caveats of FCM analysis and by highlighting prerequisites for and some examples of a successful application of such methods. Collecting faecal samples and analysing FCMs may appear simple and straightforward, but researchers have to select and apply methods correctly. They also need to be aware of the many pitfalls and potentially confounding factors and, last but not least, have to carefully interpret results. Applied properly, measurement of FCMs is a powerful non-invasive tool in a variety of research areas, such as (stress) biology, ethology, ecology, animal conservation and welfare, but also biomedicine.

Abstract: Animal management often involves procedures that, while unlikely to cause physical pain, still cause aversive responses. The domestic horse (Equus caballus) regularly has excessive hair clipped off to facilitate its use as a riding/driving animal and this procedure causes adverse behavioral responses in some animals. The aim of this study was to compare behavioral and physiological measures to assess the aversive effect of this procedure. Ten horses were selected on the basis of being either compliant (C: n=5) or non-compliant (NC: n=5) during this procedure. The horses were subjected to a sham clipping procedure (SC: where the blades had been removed from the clippers) for a period of ten minutes. Measures were taken pre, during and post SC (-10min to +30min) and mean values calculated for ALL horses and for C and NC separately. Behavioral activity was scored (scale 1-5) by twenty students from video footage in (phase/group-blind scoring). Heart rate (HR), salivary cortisol and eye temperature were monitored throughout the procedure. The NC horses were found to be significantly more behaviorally active/less relaxed throughout the trial than C horses (p<0.05) with the greatest difference occurring during the SC procedure (p<0.01). NC horses were more active/less relaxed during, compared with pre or post SC (p<0.05), but showed no behavioral difference pre and post SC. HR of the NC horses was higher than that of the C horses throughout the trial but only significantly so after 10min of SC (p<0.01). ALL horses showed a significant increase in HR between +5 and +10min into the procedure (p<0.05). There was a significant increase in salivary cortisol concentration in ALL horses post procedure (p<0.01) with levels peaking at 20minute post SC. No significant differences in salivary cortisol concentration between C and NC were found at any stage of the trial. Eye temperature increased significantly in ALL horses during SC, peaking at +10min into the procedure (p<0.05) and then decreased substantially when SC had ceased (p<0.01). Although no significant differences were found between C and NC per se, there was a significant interaction between group and phase of trial (p<0.05) with the NC group showing a greater decrease in eye temperature post SC. There was a significant positive correlation between changes in salivary cortisol concentration and eye temperature (p<0.01) but no correlation between any of the other measures. Although the behavioral response of C and NC to this procedure was significantly different the physiological responses indicated that ALL horses found the procedure aversive. Eye temperature could be used as an objective and immediate measure of how an animal is responding to a specific situation in order to evaluate management procedures and adapt them where appropriate to reduce the negative impact on animal health and welfare.

Abstract: Predation on livestock is a source of human-wildlife conflicts and can undermine the conservation of large carnivores. To design effective mitigation strategies, it is important to understand the determinants of predation across livestock species, which often differ in husbandry practices, vulnerability to predators and economic value. Moreover, attention should be given to both predation occurrence and intensity, because these can have different spatial patterns and predictors. We used spatial risk modelling to quantify factors affecting wolf predation on five livestock species in Portugal. Within the 1619 parishes encompassing the entire wolf range in the country, the national wolf compensation scheme recorded 17,670 predation events in 2009-2015, each involving one or more livestock species: sheep (31.7%), cattle (27.7%), goats (26.8%), horses (14.8%) and donkeys (3.2%). Models built with 2009-2013 data and validated with 2014-2015 data, showed a shared general pattern of predation probability on each species increasing with its own density and proximity to wolf packs. For some species there were positive relations with the density of other livestock species, and with habitat variables such as altitude, and land cover by shrubland and natural pastures. There was also a general pattern for predation intensity on each species increasing with its own density, while proximity to wolf packs had no significant effects. Predation intensity on goats, cattle and horses increased with the use of communal versus private pastures. Our results suggest that although predation may occur wherever wolves coexist with livestock species, high predation intensity is mainly restricted to particular areas where husbandry practices increase the vulnerability of animals, and this is where mitigation efforts should concentrate.

Abstract: The social and spatial organization of urban fox groups prior to and during an outbreak of sarcoptic mange was compared with predictions derived from the resource dispersion hypothesis (RDH). We investigated the availability of three key resources. Neither daytime rest sites nor breeding sites appeared to be limited in availability. The availability of food deliberately supplied by local householders was examined by questionnaire surveys. The daily and weekly amount of food supplied was greatly in excess of the minimum requirements of a pair of foxes, but was consistent between territories. The availability of this food source increased markedly as a result of more people feeding the foxes. In agreement with the RDH, group size prior to the outbreak of mange increased from 2.25 animals (N=4) to 6.57 animals (N=7). Before the outbreak of mange, two territories were divided. Increased scavenge availability on smaller territories may have promoted these changes. Excluding these spatial changes, territories were very stable between years. After the outbreak of mange, group size declined as a direct result of mange-induced mortality. Surviving animals increased their ranges only after neighbouring groups had died out. Ranges did not increase in size in response to a decline in food availability. Nor were the increases in range size associated with the relinquishment of parts of the existing territory. These postmange changes are contrary to the RDH. Three factors may have promoted these changes: the elimination of interstitial space, the forced dispersal of young or future division of the territory.