Appeasing pheromones for the management of stress and aggression during conservation of wild canids: could the solution be right under our nose?

Thirty-six species of canid exist globally, two are classified as critically endangered, three as endangered, and five as near threatened. Human expansion and the coinciding habitat fragmentation necessitate conservation interventions to mitigate concurrent population deterioration. The current conservation management of wild canids includes animal translocation and artificial pack formation. These actions often cause chronic stress, leading to increased aggression and the suppression of the immune and reproductive systems. Castration and pharmaceutical treatments are currently used to reduce stress and aggression in domestic and captive canids. The undesirable side effects make such treatments inadvisable during conservation management of wild canids. Pheromones are naturally occurring chemical messages that modulate behaviour between conspecifics; as such, they offer a natural alternative for behaviour modification. Animals are able to distinguish between pheromones of closely related species through small compositional differences but are more likely to have greater responses to pheromones from individuals of the same species. Appeasing pheromones have been found to reduce stress- and aggression-related behaviours in domestic species, including dogs. Preliminary evidence suggests that dog appeasing pheromones (DAP) may be effective in wild canids. However, the identification and testing of species-specific derivatives could produce more pronounced and beneficial behavioural and physiological changes in target species. In turn, this could provide a valuable tool to improve the conservation management of many endangered wild canids

Preliminary findings on the effect of the pig appeasing pheromone in a slow releasing block on the welfare of pigs at weaning

The pig appeasing pheromone (PAP) applied in spray has shown to be effective in reducing the frequency of aggression and the stress response of young and adult pigs under experimental conditions. This preliminary experiment investigates the effect of the PAP in a slow releasing block on the behaviour and skin lesions of weaners after mixing on a commercial farm. Two identical rooms containing six replicates per treatment of a commercial weaner building were used. There were two treatments (PAP block and Control) and each room contained one treatment. In the PAP treatment, the day before weaning and after washing and disinfection, two PAP blocks (1 block / 20 m 2) were placed hanging on top of the pens. Six hours after mixing, there was a significant effect of the PAP block treatment on the occurrence of social negative behaviour (PAP mean; median [95 % CI] for median = 3.6; 2.7 [1.5-5.4] % vs. Control 8.3; 8.7 [6.1-11.3] % P = 0.003, social positive behaviour (PAP 5.5; 5.1 [4.5-6.8] % vs. Control 1.4; 1.3 [0-2.8] % P = 0.02), feeding (PAP 9.1; 7.4 [5.5-16.5] % vs. Control 1.6; 0.0 [0-6.1] % P = 0.02, drinking (PAP 3.1; 2.9 [2.5-3.9] % vs. Control 0.8; 0.5 [0-1.7] % P = 0.04) as well as on the sitting position (PAP 1.2; 0.9 [0-2.4] % vs. Control 4.1; 4.2 [2.1-6.0] % P = 0.02). Except for social positive behaviour (PAP 5.6; 6.0 [3.9-7.5] % vs. Control 2.9; 2.8 [1.6-4.2] % P = 0.02) those differences were not significant 24 h post-mixing anymore. The prevalence of wounded animals was not significantly different between treatments. The PAP in block form may be a promising tool to reduce food neophobia and aggression after mixing. Still, further research is needed to increase the effect of the PAP block over time under commercial farming conditions

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

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

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

Dog appeasing pheromone prevents the androgen surge and may reduce contact dominance and active submission after stressful interventions in African wild dogs (Lycaon pictus)

The endangered African wild dog (AWD; Lycaon pictus) is a highly social canid living in packs with a separate male and female hierarchy. Immobilisation, handling and translocations are acute stressors for AWDs, however such interventions are often needed for species management. In addition, new pack formation or temporary pack separation can lead to an increase in intra-pack aggression. The goal of this double-blinded placebo-controlled study conducted in captive zoo populations was to evaluate whether dog appeasing pheromone (DAP) reduces behavioural stress and faecal glucocorticoid metabolite levels (fGCM) normally associated with pack separation, immobilisation and reintroduction (SIR), and to assess whether this reduces aggressive behaviours and faecal androgen metabolite levels (fAM). Four packs (n = 11 males) were treated with DAP and 4 packs (n = 12 males) were treated with a placebo solution, applied at the end of anaesthesia. Behavioural interactions as well as fGCM and fAM were determined from 3 days before until 4–6 days after SIR. No effect of DAP on fGCM was observed, however, fAM increased after SIR in placebo but not DAP treated animals. Moreover, on the day of reintroduction, DAP treated packs tended to have lower rates of contact-dominance and active-submission behaviour, but higher rates of non-contact dominance behaviour. As these effects could decrease the risk of agonistic interactions, DAP may be a useful tool to help manage new pack formations and temporary pack separation

Pathology and behaviour in feline medicine: investigating the link between vomeronasalitis and aggression

Objectives The aim of the study was to investigate if the feline vomeronasal organ (VNO) can be affected by inflammatory lesions and if these changes are associated with behavioural alterations. Methods VNOs from 20 cats were sampled during necropsy, submitted for routine tissue processing and stained with haematoxylin and eosin for histopathological evaluation. Of the 20 cats, data on the presence of aggressive behaviours towards cats or humans were collected by questionnaire survey at the point of death. Inflammatory lesions were classified depending on the duration of the process as acute or chronic, both in vomeronasal sensory epithelium (VNSE) and in non-sensory epithelium (NSE). Fisher’s exact test was used to compare VNO inflammation with behavioural data. Results The VNSE was inflamed in 11/20 VNOs (55%) while the NSE was inflamed in 13/20 (65%). Overall, the VNO was affected by inflammation in 14/20 (70%) cats, and all the lesions were classified as chronic. Five out of 20 cats (25%) had documented intraspecific aggressive behaviours and 8/20 (40%) had shown aggression towards humans. Fisher’s exact test showed a statistically significant correlation between inflammation of the VNSE and intraspecific aggression (P = 0.038). No statistically correlations were observed between VNSE inflammation and aggression towards humans and between NSE inflammation and aggression towards cats or humans. Conclusions and relevance Our results show, for the first time, the existence of vomeronasalitis in animals and its possible association with intraspecific aggressive behaviours. The inflammatory microenvironment could impair VNSE functionality, causing intraspecific communication alterations, probably through a reduction in chemical communication action and perception. Owing to the pivotal role of the VNO in the social life of cats and other species, this report provides a rationale to further investigate this disease in relation to a variety of behavioural disorders

Novel appeasing pheromones to minimize stress during metapopulation management of African wild dogs

In the last century, human persecution and increased habitat fragmentation has reduced African wild dog populations from around 500,000 to 6,600 individuals, with the population continuing to decline. In South Africa, metapopulation management has been instrumental to the survival of populations across fragmented habitats, and entails the translocation and artificial new pack formation of animals to reinforce populations, maximize genetic dispersal and reintroduce animals into their former range. However, temporary captivity during such conservation interventions regularly cause chronic stress in African wild dogs, and can result in increased aggression, disrupted pack hierarchy, injury and occasionally mortality. Pheromones are naturally occurring chemicals that can moderate behaviours and physiology in conspecifics. Appeasing pheromones identified in domestic dogs (DAP) are known to reduce stress and aggression. When applied to African wild dog packs, we showed DAP treatment decreased faecal androgen metabolite concentrations and shifted dominance behaviour from contact to non-contact compared to controls (Van den Berghe et al. PLoS ONE 14(3): e0212551). However, because pheromones are largely species-specific, an African wild dog-specific appeasing pheromone (AWDAP) should elicit a stronger beneficial physiological and behavioural effect during conservation interventions. In this project, AWDAP will be isolated and applied to packs undergoing translocation in South Africa. Using a combination of behaviour, faecal hormone metabolites and antibodies, animals will be monitored for reduced stress and aggression, improved immune function and greater social cohesion, all of which should ultimately confer a survival advantage to the pack upon release into the wild

Molecular docking of ABP dimers with steroids.

<p>The 2D-residue map of steroid interaction with ABP dimers; the residues within 4.0 Å of the steroids are shown. (A-C). Residue interactions of progesterone (P), testosterone (T), and dihydrotestosterone (DHT) with the AB dimer. (D-F). Residue interactions of progesterone, testosterone, and dihydrotestosterone with the AG dimer. The protein-ligand interactions are depicted as Van der Waals, conventional hydrogen bonds, and alkyl bonds.</p

Legal Frameworks and Controls for the Protection of Research Animals: A Focus on the Animal Welfare Body with a French Case Study

In recent years, efforts have been devoted to improving the welfare of laboratory animals. Scientific progress and growing concerns over animal harm have pushed institutions to strengthen their laws to make science more humane and responsible. European Directive 2010/63/EU makes it mandatory for breeders, suppliers and users of laboratory animals to have an animal welfare body (AWB) to prioritize animal welfare and harmonize experimental standards while reassuring the public that research is being carried out appropriately. Based on application of the three Rs (refinement, reduction and replacement), these bodies provide staff with oversight and advisory functions to support compliance with the legal requirements on both animal housing and project realization. This review aims to present the legal measures protecting research animals, with a focus on European AWBs. The review explains how the mission of AWBs includes development of environmental enrichment programs and how animal training generates benefits not only for animal welfare but also for the research work environment and research quality. A French case study is conducted to provide the scientific community with an example of an AWB’s functioning and activities, share its achievements and propose some perspectives for the future