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.

Abstract: Intraspecific allometric calculations of the brain to body size relation revealed distinct differences between 127 (67; 60) ancestral wild cavies and 82 (37; 45) guinea pigs, their domesticated relatives. The dependency of both measures from one another remained the same in both animal groups but the brains of guinea pigs were by 14.22% smaller at any net body weight. Consistent with results in other species the domestication of Cavia aperea is also characterized by a decrease of brain size. Fresh tissue sizes of the five brain parts medulla oblongata, cerebellum, mesencephalon, diencephalon and telencephalon were determined for 6 cavies and 6 guinea pigs by the serial section method. Additionally the sizes of 16 endbrain structures and those of the optic tract, the lateral geniculate body and the cochlear nucleus were measured. Different decrease values resulted for all these structures concomitant with domestication as was calculated from the amount of total brain size decrease and average relative structure values in the wild as well as the domesticated brain. The size decrease of the entire telencephalon (-13.7%) was within the range of the mean overall reduction as similarly was the case for the total neocortex (-10.7%) whereas the total allocortex (-20.9%) clearly was more strongly affected. The size decrease of the olfactory bulb (-41.9%) was extreme and clearly higher than found for the secondary olfactory structures (around -11%). The primary nuclei of other sensory systems (vision, audition) were decreased to less extent (lateral geniculate: -18.1%; cochlear nucleus: -12.6%). Mass decreases of pure white matter parts were nearly twice as high in contrast to associated grey matter parts (neocortex white versus grey matter; tractus opticus versus lateral geniculate body). The relatively great decrease values found for the limbic structures hippocampus (-26.9%) and schizocortex (-25.9%) are especially notable since they are in good conformity with domestication effects in other mammalian species. The findings of this study are discussed with regard to results of similar investigations on wild and domesticated gerbils (Meriones unguiculatus), the encephalization of the wild form, the special and species-specific mode and duration of domestication and in connection with certain behavioral changes as resulted from comparative investigations in ethology, socio-biology, endocrinology and general physiology.

Abstract: The first month of training of a young horse is suspected to be stressful, but the endocrine responses to initial training are unknown. Therefore in our study a total of 40 Quarter Horses (QH), all at the age of almost 2 years, were followed during the first 30 days of their training. During this time faecal samples were collected twice daily and faecal cortisol metabolites (FCM) were measured. Baseline values of FCM ranged between 1.3 and 20.1 (median: 6.7) ng/g faeces. No differences in FCM values between days of training were found. Mares showed the highest values. Significant diurnal variations were observed in mares (p=0.035) and stallions (p=0.003), but not in geldings (p=0.282). As in this study adrenocortical activity was not increased during initial training, horses seem to cope very well with this new situation. The results of our large-scale study provide basic physiological data about initial training. This gives additional input in an emotional debate about animal welfare aspects of first time handling and training of horses.
Abbreviations: 11,17-DOA = 11,17-dioxoandrostanes; EIA = Enyzme Immunoassay; FCM = faecal cortisol metabolites; GC = glucocorticoids; HPA-axis = hypothalamic-pituitary-adrenocortical-axis; QH = Quarter Horses
[Das Einreiten eines jungen Pferdes steht unter Verdacht belastend zu sein. Bisher gibt es aber keine Veröffentlichungen über endokrine Vorgänge während dieser Phase. Mit der vorliegenden Studie wurde überprüft, ob Pferde aufgrund physischer und psychischer Belastungen während des Trainings höhere Konzentrationen an Kortisolmetaboliten im Kot (FCM) aufweisen. Es wurden dazu 40 Quarter Horses im Alter von 2 Jahren während der ersten 30 Tage der Grundausbildung des Westernreitens beobachtet und ihre FCM Werte gemessen. Während dieser Zeitspanne wurden täglich morgens und abends Kotproben der Pferde genommen. Die Basalwerte der FCM Konzentration variierten zwischen 1,3 und 20,1 (Median: 6,7) ng/g Kot, wobei Stuten die höchsten Werte hatten. Signifikante Unterschiede während der einzelnen Trainingstage konnten nicht festgestellt werden. In der Tagesrhythmik wurden signifikante Unterschiede bei Stuten (p=0,035) und bei Hengsten (p=0,003), jedoch nicht bei Wallachen (p=0,282) ermittelt. In dieser Studie konnte keine erhöhte Aktivität der Nebennierenrinde im Verlauf der Grundausbildung eines Pferdes im Westernreitstil festgestellt werden. Das legt nahe, dass Pferde mit dieser neuen, zeitlich kurz andauernden Situationen gut zurechtkommen. Unsere Studie wurde an einer großen Anzahl von Tieren unter Feldbedingungen durchgeführt. Sie bietet daher eine gute Datenbasis über Belastungen während des Einreitens. Damit liefert sie einen zusätzlichen Beitrag zu einer mitunter emotional geführten Debatte über tierschutzrelevante Aspekte bei der Grundausbildung von Pferden.]

Abstract: Starting with dogs, over 15,000 years ago, the domestication of animals has been central in the development of modern societies. Because of its importance for a range of disciplines – including archaeology, biology and the humanities – domestication has been studied extensively. This chapter reviews how the field of paleogenomics has revolutionised, and will continue to revolutionise, our understanding of animal domestication. We discuss how the recovery of ancient DNA from archaeological remains is allowing researchers to overcome inherent shortcomings arising from the analysis of modern DNA alone. In particular, we show how DNA, extracted from ancient substrates, has proven to be a crucial source of information to reconstruct the geographic and temporal origin of domestic species. We also discuss how ancient DNA is being used by geneticists and archaeologists to directly observe evolutionary changes linked to artificial and natural selection to generate a richer understanding of this fascinating process.