Abstract: The traditional way to train horses is by the application of negative reinforcement (NR). In the past few years, however, the use of positive reinforcement (PR) has become more common. To evaluate the effectiveness and the possible stressor effect of the 2 training methods, 12 horses showing severe trailer-loading problems were selected and exposed to trailer-loading. They were randomly assigned to one of the 2 methods. NR consisted of various degrees of pressure (lead rope pulling, whip tapping). Pressure was removed as soon as the horse complied. PR horses were exposed to clicker training and taught to follow a target into the trailer. Heart rate (HR) was recorded every 5 seconds and behavior denoting discomfort was observed using one-zero sampling with 10 seconds sampling intervals. Training was completed when the horse could enter the trailer upon a signal, or was terminated after a maximum of 15 sessions. Of the 12 horses, 10 reached the criterion within the 15 sessions. One horse was eliminated from the study because of illness and 1 PR horse failed to enter the trailer. A Mann-Whitney U-test indicated that the horses trained with NR displayed significantly more discomfort behavior per training session than horses trained with PR (NR: 13.26 ± 3.25; PR: 3.17 ± 8.93, P < 0.0001) and that horses in the PR group spent less time (second) per session to complete the training criterion (NR: 672.9 ± 247.12; PR: 539.81 ± 166.37, P < 0.01). A Mann-Whitney U-test showed that no difference existed in mean HR (bpm) between the 2 groups (NR: 53.06 ± 11.73 bpm; PR: 55.54 ± 6.7 bpm, P > 0.05), but a Wilcoxon test showed a difference in the PR group between the baseline of HR and mean HR obtained during training sessions (baseline PR: 43 ± 8.83 bpm; PR: 55.54 ± 6.7 bpm, P < 0.05). In conclusion, the PR group provided the fastest training solution and expressed less stress response. Thus, the PR procedure could provide a preferable training solution when training horses in potentially stressing situations.

Abstract: The ability to innovate and the social transmission of innovations have played a central role in human evolution. However, innovation is also crucial for other animals, by allowing them to cope with novel socioecological challenges. Although innovation plays such a central role in animals' lives, we still do not know the conditions required for innovative behaviour to emerge. Here, we focused on interindividual differences in innovation by (1) extensively reviewing existing literature on innovative behaviour in animals and (2) quantitatively testing the different evolutionary hypotheses that have been proposed to explain interindividual variation in innovation propensity during foraging tasks. We ran a series of phylogenetically controlled mixed-effects meta-regression models to determine which hypotheses (if any) are supported by currently available empirical studies. Our analyses show that innovation is more common in individuals that are older and belong to the larger sex, but also in more neophilic and/or explorative individuals. Moreover, these effects change depending on the study setting (i.e. wild versus captive). Our results provide no clear support to the excess of energy or the bad competitor hypotheses and suggest that study setting and interindividual differences in traits related to personality are also important predictors of innovation.

Abstract: Developmental changes in time spent suckling and related mother-foal behaviour are described in an unmanaged herd of Camargue horses. Male foals spent about 40% more time suckling than females during the first 8 weeks. Body weight did not differ between the sexes but time-budgets did: males grazed less and were more active. If pregnant, the typical multiparous mare nursed her foals for 35–40 weeks, males and females alike, and weaned them 15 weeks before the next foaling. Primiparae lactated longer and weaned closer to the next foaling by 5 weeks. The mares played an active role in regulating the time spent suckling in early, and particularly in late lactation.

Abstract: An immunoglobulin G (IgG) was spray-dried on a Büchi 190 laboratory spray-dryer at inlet and outlet air temperatures of 130 and 190°C, respectively. The IgG solution contains initially 115mg/ml IgG plus 50mg/ml sorbitol. After dialysis, at least 80% of low molecular weight component was removed. After spray-drying the dialyzed IgG and immediate redissolution of the powder, an increase in aggregates from 1 to 17% occurred. A major shift towards increase β-sheet structure was detected in the spray-dried solid, which, however, reverted to native structure on redissolution of the powder. A correlation between aggregation determined by size exclusion chromatography and alterations in secondary structure determined by Fourier transformation infra-red spectroscopy could not therefore be established. On spray-drying a non-dialyzed, sorbitol-containing IgG only some 0.7% aggregates were formed. The sorbitol is therefore evidently able to stabilize partially the IgG during the process of spray-drying. Addition of trehalose to the liquid feed produced quantitatively the same stabilizing action on the IgG during spray-drying as did the sorbitol. This finding again points towards a water replacement stabilization mechanism. The IgG spray-dried powder prepared from the dialyzed liquid feed showed continued substantial aggregation on dry storage at 25°C. This was substantially less in the non-dialyzed, sorbitol-containing spray-dried powder. Addition of trehalose to both dialyzed and non-dialyzed system produced substantial improvement in storage stability and reduction in aggregate formation in storage. The quantitative stabilizing effect of the trehalose was only slightly higher than that of the sorbitol. Taken together, these results indicate that both the sorbitol and trehalose stabilize the IgG primarily by a water replacement mechanism rather than by glassy immobilization. The relevance of this work is its questioning of the importance of the usually considered dominance of glassy stabilization of protein in dried systems of high glass transition temperature, such as trehalose. The low glass transition temperature sorbitol produces almost equal process and storage stability in this case.

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.