Abstract: Learning through the observation of others allows the transfer of information without the costs incurred during individual trial and error learning. Horses (Equus caballus) are a highly social species, which might be expected to be capable of learning from others, but experimental findings are inconsistent, and potentially confounded by social facilitation effects not related directly to the learning of the task. We refined the methods used in previous equine social learning studies, to examine and distinguish specific social influences on learning of a task: we used predefined group leaders rather than agonistically dominant individuals to demonstrate a detour task to familiar conspecific observers; in addition we had two control groups: a non-observer (true control) and a group with the demonstrator simply present at the goal (social facilitation control). 44 socially kept horses were allocated to one of the three test conditions and took part in five trials each. Success rate, latency and detour direction were recorded. There was no significant difference between the three groups in the likelihood of them succeeding in the task nor latency to succeed; however there was a significant difference in the route chosen by the groups, with the true control choosing the side with the entrance gate significantly more than either the observer group or social facilitation group. Both of the latter two groups chose to go in the same direction relative to themselves, regardless of which side the gate was. Seven out of nine horses in the observer group chose the same direction as their demonstrator every time. Our results show a significant role of social facilitation on detour behaviour and highlight the importance of including adequate controls for simpler cognitive influences on behaviour before claims can be made about the specific learning of motor actions or goal directed behaviour. Social cues may be important to horses if the task is sufficiently challenging and motivationally important, so future work should consider more demanding, but ecologically relevant situations, in order to maximise the potential revelation of social learning effects which do not depend on simple local or stimulus enhancement effects.

Abstract: Social licking patterns of heifer and steer herds were observed and recorded during periods of resting and intermittent feeding. The results revealed the following features: (1) heifers and steers had 15.0 and 15.2 social licking interactions per hour which lasted for 37.8 and 41.0 s on average, respectively. The average time an animal spent licking was about 25 s per hour; (2) all the animals in the herds were licked by others, but only 72.3% of the animals licked other animals; (3) the animals close in the social hierarchy tended to lick each other for a longer time than did remote animals; (4) the time receiving l licking and weight gain tended to be positively correlated. The observations suggest that (1) the motivation of giving licking may be individual-specific and may be influenced by genetic factors, while that of receiving licking appears to be general, and that (2) social licking may mean not only cleaning the skin and hair of a passive partner, but also leading it to psychological stability.

Abstract: Monitoring of the wolf Canis lupus is a demanding task as it lives in low densities, utilizes vast home ranges and disperses over large areas. These factors make obtaining accurate data about population parameters over the whole distribution area of the species impossible. Thus detailed local studies on socio-spatial organization are essential to calibrate information obtained over a larger area. We applied GPS/GSM telemetry, non-invasive genetic sampling, year-round tracking, camera trapping and howling stimulations to determine the number of family groups, population density and home-range sizes of wolves in the Drawa Forest (DF, western Poland, 2500 km2), an area recently recolonized by the species. Home ranges of three collared male wolves ranged from 321.8 to 420.6 km2 (MCP 100%) and from 187.5 to 277.5 km2 (Kernel 95%), but core areas had a size of 30.5-84.7 km2 (MCP50%) and 35.0-88.8 km2 (Kernel 50%). Mean near neighbour distance between centres of 6 tracked pack homesites was 15.3 km. The number of wolves in DF increased from 14 individuals in 2013/2014 to 30 in 2016/2017. The annual rate of increase varied from 43% in 2014/2015 to 7% in the final year. Population density for the whole study area was relatively low (1.2 indiv./100 km2 in 2016/2017), but densities within territories of two packs studied with telemetry were 1.9 and 1.5 indiv./100 km2. Mean pack size varied between 3.5 and 5.6 individuals, with the largest pack comprising 8 wolves. Mean number of pups observed in summers (June-August) was 4.5. Differences in home range sizes between wolves in western and eastern Poland indicate that results of regional studies cannot be freely extrapolated despite close genetic relationships. Thus, decisions related to management of wolf habitats should be based on intensive local studies.

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.