Effect of wither application of an analogue of pig appeasing pheromone on encounters between unfamiliar mini-pigs.

BACKGROUND: The practice of mixing unfamiliar pigs on farms is common but results in fighting, welfare problems and performance issues. Pigs have different ways of resolving social conflicts, including aggressive and affiliative behaviours. Synthetic appeasing pheromones have demonstrated many positive effects in animal husbandry and are regularly used by breeders to improve animal welfare and performance. The aim of the study was to investigate the effect of a new method of applying pig appeasing-pheromone (PAP) to the withers in an experimental model of pig mixing to determine whether PAP reduced aggression and fighting, increased prosocial behaviours, and improved behavioural and physiological indicators of welfare. RESULTS: PAP reduced fighting between mini-pigs (df = 1; F = 13.47; P = 0.001; mixed logistic regression). Even if not significant, agonistic behaviours tended to be reduced when the treatment was applied (df = 1; F = 4.14; P = 0.058; mixed logistic regression). Likewise, mini-pigs seemed to be scored as not aggressive at all (df = 1; F = 3.61; P = 0.070; GLMM) and to be less aggressive toward the other pig than when placebo was applied. Concerning the latency of the first contact without aggression, a significant effect was found between the PAP and placebo groups (df = 1; χ2 = 4.74; P = 0.0295; Cox model). Moreover, even if not significant, the treated mini-pigs seemed to spent more time looking at each other (df = 1; F = 3.59; P = 0.071; GLMM) and immobile and/or ground sniffing (df = 1; F = 3.18; P = 0.088; GLMM) than those that received placebo. No significant difference was found between groups for salivary cortisol concentration (df = 1; F = 0.10; P = 0.752; GLMM), but variances between groups were heterogeneous at every time. No significant difference was found between groups regarding alpha-amylase activity (df = 1; F = 0.25; P = 0.621; GLMM), but variances between groups were heterogeneous at T0, T1 and T3. These results indicate that the variability (dispersion) within each group was lower when PAP was applied than when the placebo was applied. CONCLUSIONS: The new method of applying PAP improved welfare of mini-pigs (as models of domestic pigs) by reducing fighting, among other interesting results. PAP seems thus a promising biomimetic tool to enhance animal welfare in pig production systems.

Inflammation interferes with chemoreception in pigs by altering the neuronal layout of the vomeronasal sensory epithelium.

Chemical communication is widely used by animals to exchange information in their environment, through the emission and detection of semiochemicals to maintain social organization and hierarchical rules in groups. The vomeronasal organ (VNO) is one of the main detectors of these messages, and its inflammation has been linked to behavioral changes because it potentially prevents molecule detection and, consequently, the translation of the signal into action. Our previous study highlighted the link between the intensity of vomeronasal sensory epithelium (VNSE) inflammation, probably induced by farm contaminant exposure, and intraspecific aggression in pigs. The aim of this study was to evaluate the cellular and molecular changes that occur during vomeronasalitis in 76 vomeronasal sensorial epithelia from 38 intensive-farmed pigs. Histology was used to evaluate the condition of each VNO and classify inflammation as healthy, weak, moderate, or strong. These data were compared to the thickness of the sensorial epithelium and the number of type 1 vomeronasal receptor cells using anti-Gαi2 protein immunohistochemistry (IHC) and analysis. The presence of odorant-binding proteins (OBPs) in the areas surrounding the VNO was also analyzed by IHC and compared to inflammation intensity since its role as a molecule transporter to sensory neurons has been well-established. Of the 76 samples, 13 (17%) were healthy, 31 (41%) presented with weak inflammation, and 32 (42%) presented with moderate inflammation. No severe inflammation was observed. Epithelial thickness and the number of Gαi2+ cells were inversely correlated with inflammation intensity (Kruskal-Wallis and ANOVA tests, p < 0.0001), while OBP expression in areas around the VNO was increased in inflamed VNO (Kruskal-Wallis test, p = 0.0094), regardless of intensity. This study showed that inflammation was associated with a reduction in the thickness of the sensory epithelium and Gαi2+ cell number, suggesting that this condition can induce different degrees of neuronal loss. This finding could explain how vomeronasalitis may prevent the correct functioning of chemical communication, leading to social conflict with a potential negative impact on welfare, which is one of the most important challenges in pig farming.

Neurohormonal Profiles of Assistance Dogs Compared to Pet Dogs: What Is the Impact of Different Lifestyles?

Assistance dogs must manage stress efficiently because they are involved in challenging tasks. Their welfare is currently a fundamental issue. This preliminary study aimed to compare assistance dogs (AD; n = 22) with pet dogs (PD; n = 24), using blood neuromodulator indicators to help find biomarkers that can improve the AD breeding, selection, training, and welfare monitoring. Both populations originated from different breeds, are of different ages, and had different lifestyles. Basal peripheral concentrations of prolactin (PRL), serotonin (5-HT), free (fOT) and total (tOT) oxytocin were measured by immunoassays. Multiple linear regressions were performed to assess the effect of activity, age, sex, and their interactions on these parameters. Correlations between neurohormonal levels were analyzed. No interactions were significant. fOT and tOT concentrations were significantly influenced by age (p < 0.0001 and p = 0.0002, respectively) and dogs' activity (p = 0.0006 and p = 0.0277, respectively). A tendency was observed for age effect on PRL (p = 0.0625) and 5-HT (p = 0.0548), as well as for sex effect on tOT (p = 0.0588). PRL concentrations were heterogenous among AD. fOT and tOT were significantly but weakly correlated (Pearson's r = 0.34; p = 0.04). Blood prolactin, serotonin, and oxytocin may represent biomarkers to assess workload and chronic stress-related responses in ADs and eventually improve their selection and training.

Early Evaluation of Fearfulness in Future Guide Dogs for Blind People.

Fear is the leading cause of guide dog failure. Detecting the nature and causes of these fears as early as possible is the first step in preventing their occurrence. The process of habituation is a fundamental part of fear prevention. In this study, 11 puppies, all five months of age, underwent an emotional reactivity test (ERT) composed of 12 scored items, classified into three categories: unknown person (UP), sound and visual stimuli (SVS), and body sensitivity (BS). Salivary cortisol was also measured. Foster families were asked to complete a questionnaire concerning puppies' habituation. The physiological data were correlated with UP (r = 0.71) and BS scores (r = 0.67), but not with SVS scores (r = 0.16), suggesting the ability of these dogs to control themselves when faced with the latter stimulus category. Additionally, the more time a puppy spent alone, the more likely it was to be afraid of SVS (p = 0.05). A correlation, albeit moderate, was detected between cortisol and habituation scores (r = 0.48). These results give us interesting avenues to explore, particularly regarding the importance of focusing on early puppy socialization and habituation to improve the numbers of guide dog candidates becoming successful guide dogs.

Effect of handling and crowding on the susceptibility of Atlantic salmon (Salmo salar L.) to Lepeophtheirus salmonis (Krøyer) copepodids.

Lepeophtheirus salmonis is an ectoparasite causing economic concerns in Atlantic salmon farming. Salmon lice infestation management methods can be stressful and impact fish welfare. This work investigated the stress effect on the attachment of L. salmonis copepodids to Atlantic salmon through two approaches: (a) handling by netting and air exposure (acute stress), and (b) crowding with restricted surface access in a tank (chronic stress). In the first experiment, we compared the number of attached L. salmonis and cortisol levels between a group of handled salmon and a control group. In the second experiment, a group of densely packed salmon was compared to a control group based on the number of attached copepodids, cortisol levels and neutrophil:lymphocyte ratios. Handled salmon showed significantly higher plasma cortisol levels (p < .001) and more attached copepodids (p = .01) than control salmon. Conversely, the cortisol level and number of attached copepodids were not significantly different between the densely packed and control salmon (p > .05). The neutrophil:lymphocyte ratio was significantly higher (p = .0014) in the densely packed salmon than in the control salmon. Handling salmon increased their risk of infestation by L. salmonis. This has implications for reinfestation rates following delousing treatments in commercial salmon aquaculture, which often involve crowding and handling salmon.

Working Smarter Not Harder: Oxytocin Increases Domestic Dogs' (Canis familiaris) Accuracy, but Not Attempts, on an Object Choice Task.

The neuropeptide oxytocin (OT) has been shown to enhance dogs' ability to perform an object choice task (OCT) involving the use of human pointing cues, when delivered intranasally. This study aimed at further investigating whether OT enhances task performance by increasing choices made, or by increasing correctness of choices made, and to compare these treatment effects to dog appeasing pheromone (DAP), known to balance emotional activation in dogs. Hence, we compared OCT performance between three groups of dogs: (i) dogs administered OT and a sham collar, (ii) dogs administered a saline placebo and a DAP collar, and (iii) control dogs administered a saline placebo and a sham collar. All three groups consisted of a combination of male and female pet dogs and assistance-dogs-in-training currently living with a volunteer carer. The study also evaluated the effect of intranasal OT and/or DAP on plasma levels of OT, and prolactin; which has previously been linked with anxiety in dogs. The dogs' emotional state was measured using the Emotional Disorders Evaluation in Dogs (EDED) scale. The owners'/carers' degree of anxious- and avoidant-style attachment to their dogs was accessed using the Pet Attachment Questionnaire (PAQ). Interesting descriptive data appeared for both treatment groups. Particularly, in OT group, we obtained significant results demonstrating that intranasal OT enhances OCT performance in dogs compared to control, by increasing the percentage of correct choices, but not the number of choices, made. Results also support that the mode of action of intranasal OT is via direct access to the brain and not via the blood, since no elevation of plasma OT (or prolactin) levels were observed after intranasal administration in this study. Similarly, DAP application did not significantly alter OT or prolactin peripheral concentrations. Several differences were observed between fostered and pet dogs, namely: fostered dogs demonstrated higher levels of serum prolactin, made more choices on the OCT compared to pet dogs but were not more likely to be correct, and were fostered by carers with higher avoidant attachment scores than pet dog owners. These findings implicate consideration of potential carer and training consequences for assistance dogs.

Assessment of Commercially Available Immunoassays to Measure Glucocorticoid Metabolites in African Grey Parrot (Psittacus Erithacus) Droppings: A Ready Tool for Non-Invasive Monitoring of Stress.

Despite being undomesticated, African Grey Parrots (Psittacus erithacus) are commonly found in captivity, in zoos or as pets. Captivity can be an ongoing stressor. Non-invasive glucocorticoid metabolites (GCM) measurements from bird droppings are of interest for assessing stress but require careful evaluation in each newly studied species. This study describes the assessment of such methods for Psittacus erithacus to provide tools for evaluating stress and monitoring welfare. We evaluated 12 method combinations of GCM extraction and enzyme immunoassay (EIA) from a pool of African Grey Parrot droppings, through the validation of several analytical parameters. Then, Heterophil: Lymphocyte Ratios (HLR), another reliable stress marker, were determined and correlated to individual dropping GCM concentrations for 29 birds to determine whether the method is biologically relevant. We found that the best procedure to measure GCM in African Grey Parrot droppings is a combination of 60% methanol extraction measured using a Corticosterone EIA kit (Cayman Chemical Company) from fresh or dry droppings. The establishment of a significant correlation (Pearson coefficient correlation = 0.48; p = 0.0082) between HLR and GCM in the studied population confirmed the method biological relevance. This method can thus be applied to assess stress in Psittacus erithacus and support welfare monitoring in zoo populations.

The effects of a synthetic analogue of the Bovine Appeasing Pheromone on milk yield and composition in Valdostana dairy cows during the move from winter housing to confined lowland pastures.

This Research Communication describes the effects of a synthetic analogue of the Bovine Appeasing Pheromone (BAP) on milk parameters in Valdostana dairy cows during the first turning out from tie-stalls to confined lowland pastures around the farms. Thirty healthy lactating Valdostana cows were enroled in the study and randomly divided into 2 groups: experimental group (EG, n = 15) and control group (CG, n = 15). The two groups were separately housed in the same farm and managed outside in two different pens. Treatment (BAP and solution) and control (solution only) were poured on the nuchal skin area between the horns when the animals were inside the farm at the feeding rack every 7 d for 28 d (T0-T4). Milk samples were evaluated at the same time points (T0-T4). Daily milk production (kg/day) was higher in the EG than in the CG, particularly during the first day after the turning out to pasture (T1). Somatic Cell Count (103 cells/ml) was higher in the placebo group than in the EG, especially at T1. Proteins, fat, fat-free dry matter and casein (g/100 g) were not affected by the treatment. In T1 urea (mg/dl) content was higher in CG vs. EG, suggesting a more correct metabolic balance in the group treated with BAP. The use of BAP appears to modulate adaptation in ways that may improve dairy cow performance in the context of changes in management routines.

Isolation and characterization of olfactory ecto-mesenchymal stem cells from eight mammalian genera.

BACKGROUND: Stem cell-based therapies are an attractive option to promote regeneration and repair defective tissues and organs. Thanks to their multipotency, high proliferation rate and the lack of major ethical limitations, "olfactory ecto-mesenchymal stem cells" (OE-MSCs) have been described as a promising candidate to treat a variety of damaged tissues. Easily accessible in the nasal cavity of most mammals, these cells are highly suitable for autologous cell-based therapies and do not face issues associated with other stem cells. However, their clinical use in humans and animals is limited due to a lack of preclinical studies on autologous transplantation and because no well-established methods currently exist to cultivate these cells. Here we evaluated the feasibility of collecting, purifying and amplifying OE-MSCs from different mammalian genera with the goal of promoting their interest in veterinary regenerative medicine. Biopsies of olfactory mucosa from eight mammalian genera (mouse, rat, rabbit, sheep, dog, horse, gray mouse lemur and macaque) were collected, using techniques derived from those previously used in humans and rats. The possibility of amplifying these cells and their stemness features and differentiation capability were then evaluated. RESULTS: Biopsies were successfully performed on olfactory mucosa without requiring the sacrifice of the donor animal, except mice. Cell populations were rapidly generated from olfactory mucosa explants. These cells displayed similar key features of their human counterparts: a fibroblastic morphology, a robust expression of nestin, an ability to form spheres and similar expression of surface markers (CD44, CD73). Moreover, most of them also exhibited high proliferation rates and clonogenicity with genus-specific properties. Finally, OE-MSCs also showed the ability to differentiate into mesodermal lineages. CONCLUSIONS: This article describes for the first time how millions of OE-MSCs can be quickly and easily obtained from different mammalian genera through protocols that are well-suited for autologous transplantations. Moreover, their multipotency makes them relevant to evaluate therapeutic application in a wide variety of tissue injury models. This study paves the way for the development of new fundamental and clinical studies based on OE-MSCs transplantation and suggests their interest in veterinary medicine.

Validation of a Commercially Available Enzyme ImmunoAssay for the Determination of Oxytocin in Plasma Samples from Seven Domestic Animal Species.

The neurohormone oxytocin (OT) has a broad range of behavioral effects in mammals. It modulates a multitude of social behaviors, e.g., affiliative and sexual interactions. Consequently, the OT role in various animal species is increasingly explored. However, several issues have been raised regarding the peripheral OT measurement. Indeed, various methods have been described, leading to assay discrepancies and inconsistent results. This highlights the need for a recognized and reliable method to measure peripheral OT. Our aim was to validate a method combining a pre-extraction step, previously demonstrated as essential by several authors, and a commercially available enzyme immunoassay (EIA) for OT measurement, using plasma from seven domestic species (cat, dog, horse, cow, pig, sheep, and goat). The Oxytocin EIA kit (EnzoLifeSciences) was used to assay the solid-phase extracted samples following the manufacturer's instructions with slight modifications. For all species except dogs and cats, concentration factors were applied to work above the kit's sensitivity (15 pg/ml). To validate the method, the following performance characteristics were evaluated using Validation Samples (VS) at various concentrations in each species: extraction efficiency via spiking tests and intra- and inter-assay precision, allowing for the calculation of total errors. Parallelism studies to assess matrix effects could not be performed because of too low basal concentrations. Quantification ranges and associated precision profiles were established to account for the various OT plasma concentrations in each species. According to guidelines for bioanalytical validation of immunoassays, the measurements were sufficiently precise and accurate in each species to achieve a total error ≤30% in each VS sample. In each species, the inter-assay precision after 3 runs was acceptable, except in low concentration samples. The linearity under dilution of dogs and cats' samples was verified. Although matrix effects assessments are lacking, our results indicate that OT plasma levels can reliably be measured in several domestic animal species by the method described here. Studies involving samples with low OT plasma concentrations should pay attention to reproducibility issues. This work opens new perspectives to reliably study peripheral OT in a substantial number of domestic animal species in various behavioral contexts.