Abstract: Laterality in any organism or species can be manifest as morphological, sensory and functional degrees of asymmetry such as hemispheric dominance, handedness or sidedness and other motor functional behaviours and as such is equally important in equitation. The influence of the horses' sex on both the direction and the degree of the laterality was explored within and between 4 experimental procedures in the 1st study. The findings showed that the direction, but not the degree of idiosyncratic motor preference in the horses was strongly sex-related. Male horses exhibited significantly more left lateralized responses and female horses exhibited significantly more right lateralized responses. Visuo-spatial ability is also likely to be important in the performance horse. In many species, moderate to large differences in visuo-spatial ability have been reported between the sexes, with superior visuo-spatial ability being reported in males of all species investigated to date. As no known studies had addressed visuo-spatial ability in the equine, the objective of the 2nd study, was to determine if visuo-spatial ability differed between male and female horses. The results produced the first behavioural demonstration of superior visuo-spatial ability in male horses, similar to that reported in other species. There is evidence to suggest that visuospatial ability and motor laterality are associated with cerebral hemispheric asymmetry and may be intrinsically linked. Brain development and laterality have also been associated with hair patterning, and, in a 3rd study we attempted to identify predictors of lateral bias in motor behaviour in horses. We investigated the relationship between the direction of facial hair whorl rotation and the incidence/direction of laterality in the horse. The findings suggest that direction of facial hair whorl rotation may be a useful indicator of lateralised motor behavioural preferences in the horse. We then attempted to establish if laterality was evident at birth in a 4th study, where we explored if neonatal foals exhibited lateralised patterns during and immediately post the birthing process that were correlated with their facial hair whorl patterns. The results showed a significant association between the sex of the foal and the choice of foreleg presented initially during 2nd stage parturition. Significantly more colt foals led with the left foreleg and significantly more filly foals led with the right foreleg than expected purely by random and the behaviour was correlated with facial hair whorl patterns. The findings also suggest that lateralisation in the horse is determined in utero as has also been shown in humans. Comparisons of wholly intact male and female horses are warranted as they might elucidate additional linkages between motor behaviour, visuo-spatial ability and brain organisation and development in the horse. Further research in this area could lead to more appropriate competition conditions (better fence design/construction on cross-country tracks) and so eliminate unnecessary levels of risk associated with many equestrian sports.

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: Changes in the secondary structure of some dozen different proteins upon lyophilization of their aqueous solutions have been investigated by means of Fourier-transform infrared spectroscopy in the amide III band region. Dehydration markedly (but reversibly) alters the secondary structure of all the proteins studied, as revealed by both the quantitative analysis of the second derivative spectra and the Gaussian curve fitting of the original infrared spectra. Lyophilization substantially increases the beta-sheet content and lowers the alpha-helix content of all proteins. In all but one case, proteins become more ordered upon lyophilization.

Abstract: Since the non-invasive field endocrinology techniques were developed, several fecal preservation and extraction methods have been established for a variety of species. However, direct adaptation of methods from previous studies for use in crested macaques should be taken with caution. We conducted an experiment to assess the accuracy and stability of fecal estrogen metabolite (E1C) and glucocorticoid metabolite (GCM) concentrations in response to several preservation parameters: (1) time lag between sample collection and fecal preservation; (2) long-term storage of fecal samples in 80% methanol (MeOH) at ambient temperature; (3) different degrees of feces drying temperature using a conventional oven; and (4) different fecal preservation techniques (i.e., freeze-drying, oven-drying, and field-friendly extraction method) and extraction solvents (methanol, ethanol, and commercial alcohol). The study used fecal samples collected from crested macaques (Macaca nigra) living in the Tangkoko Reserve, North Sulawesi, Indonesia. Samples were assayed using validated E1C and GCM enzyme immunoassays. Concentrations of E1C and GCM in unprocessed feces stored at ambient temperature remained stable for up to 8 h of storage after which concentrations of both E1C and GCM changed significantly compared to controls extracted at time 0. Long-term storage in 80% MeOH at ambient temperature affected hormone concentrations significantly with concentrations of both E1C and GCM increasing after 6 and 4 months of storage, respectively. Drying fecal samples using a conventional oven at 50, 70, and 90 °C did not affect the E1C concentrations, but led to a significant decline for GCM concentrations in samples dried at 90 °C. Different fecal preservation techniques and extraction solvents provided similar results for both E1C and GCM concentrations. Our results confirm previous studies that prior to application of fecal hormone analysis in a new species, several preservation parameters should be evaluated for their effects on hormone metabolite stability. The results also provide several options for fecal preservation, extraction, and storage methods that can be selected depending on the condition of the field site and laboratory.