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: Summary Different head?neck positions (HNPs) are used in equestrian sports and are regarded as desirable for training and competition by riders, judges and trainers. Even though some studies have been indicative of hyperflexion having negative effects on horses, this unnatural position is frequently used. In the present study, the influence of different HNPs on physical and psychological stress parameters in the ridden horse was investigated. Heart rate (HR), heart rate variability (HRV) and blood cortisol levels were measured in 18 horses. Low frequency (LF) and high frequency (HF) are power components in the frequency domain measurement of HRV which show the activity of the sympathetic and parasympathetic nervous system. Values were recorded at rest, while riding with a working HNP and while riding with hyperflexion of the horse's head, neck and poll. In addition, rideability and behaviour during the different investigation stages were evaluated by the rider and by an observer. Neither the HR nor the HRV showed a significant difference between working HNP (HR = 105 ± 22/min; LF/HF = 3.89 ± 5.68; LF = 37.28 ± 10.77%) and hyperflexion (HR = 110 ± 18; LF/HF = 1.94 ± 2.21; LF = 38.39 ± 13.01%). Blood cortisol levels revealed a significant increase comparing working HNP (158 ± 60 nm) and hyperflexion (176 ± 64 nm, p = 0.01). The evaluation of rider and observer resulted in clear changes of rideability and behavioural changes for the worse in all parameters collected between a working HNP and hyperflexion. In conclusion, changes of the cortisol blood level as a physical parameter led to the assumption that hyperflexion of head, neck and poll effects a stress reaction in the horse, and observation of the behaviour illustrates adverse effects on the well-being of horses during hyperflexion.

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 aim of this study was to conduct a series of paper-based exercises in order to assess the negative (adverse) welfare impacts, if any, of common interventions on domestic horses across a broad range of different contexts of equine care and training. An international panel (with professional expertise in psychology, equitation science, veterinary science, education, welfare, equestrian coaching, advocacy, and community engagement; n = 16) met over a four-day period to define and assess these interventions, using an adaptation of the domain-based assessment model. The interventions were considered within 14 contexts: C1 Weaning; C2 Diet; C3 Housing; C4 Foundation training; C5 Ill-health and veterinary interventions (chiefly medical); C6 Ill-health and veterinary interventions (chiefly surgical); C7 Elective procedures; C8 Care procedures; C9 Restraint for management procedures; C10 Road transport; C11 Activity—competition; C12 Activity—work; C13 Activity—breeding females; and C14 Activity—breeding males. Scores on a 1–10 scale for Domain 5 (the mental domain) gathered during the workshop were compared with overall impact scores on a 1–10 scale assigned by the same panellists individually before the workshop. The most severe (median and interquartile range, IQR) impacts within each context were identified during the workshop as: C1 abrupt, individual weaning (10 IQR 1); C2 feeding 100% low-energy concentrate (8 IQR 2.5); C3 indoor tie stalls with no social contact (9 IQR 1.5); C4 both (i) dropping horse with ropes (9 IQR 0.5) and forced flexion (9 IQR 0.5); C5 long-term curative medical treatments (8 IQR 3); C6 major deep intracavity surgery (8.5 IQR 1); C7 castration without veterinary supervision (10 IQR 1); C8 both (i) tongue ties (8 IQR 2.5) and (ii) restrictive nosebands (8 IQR 2.5); C9 ear twitch (8 IQR 1); C10 both (i) individual transport (7.00 IQR 1.5) and group transport with unfamiliar companions (7 IQR 1.5); C11 both (i) jumps racing (8 IQR 2.5) and Western performance (8 IQR 1.5); C12 carriage and haulage work (6 IQR 1.5); C13 wet nurse during transition between foals (7.5 IQR 3.75); and C14 teaser horse (7 IQR 8). Associations between pre-workshop and workshop scores were high, but some rankings changed after workshop participation, particularly relating to breeding practices. Domain 1 had the weakest association with Domain 5. The current article discusses the use of the domain-based model in equine welfare assessment, and offers a series of assumptions within each context that future users of the same approach may make when assessing animal welfare under the categories reported here. It also discusses some limitations in the framework that was used to apply the model.