Cutaneous application of SecurePig® FLASH, a Pig appeasing pheromone analogue, facilitates adaptation and manages social behavior during feeding in semi-extensive conditions.

BACKGROUND: Farm animals face several challenges throughout their lives, which can affect both their welfare and their productivity. Promoting adaptation in animals is one way of limiting these consequences. In various animal species, the use of maternal appeasing pheromones is efficient to reduce aggressiveness, improve adaptation and thus ensuring better welfare and productivity. This study sought to investigate the efficiency of a treatment with a Pig Appeasing Pheromone (PAP) on the behavior of pigs reared under semi-extensive conditions and exposed to a potential conflict- collective feeding. Animals (n = 14 divided in 2 groups of 7) were subjected to 3 different phases, (A) baseline - no pigs received the PAP, (B) SP - 2 out of the 7 pigs per group received the PAP and (C) AP- all pigs received the PAP. Behaviors related to feeding, aggression and locomotion were compared between the 3 phases of the study. RESULTS: Compared to the baseline period, we observed that the number of head knocks was reduced when some pigs (p < 0.001) and all pigs (p < 0.005) received the PAP. Similarly, we observed that the number of fleeing attempts was reduced when some pigs (p < 0.001) and all pigs (p < 0.001) were treated when compared to baseline. This number was lower in the AP phase than in the SP phase (p < 0.001). When all pigs were treated (AP), we also observed that they spent less time investigating the floor than during the two other phases (p < 0.001), but they seemed more likely to leave the feeder due to the presence or behavior of another pig of the group (SP vs. AP, p < 0.05). CONCLUSIONS: The PAP application improved adaptation in pigs by reducing aggressiveness and promoting conflict avoidance. Those results validate the efficiency of the pheromonal treatment under semi-extensive rearing conditions to help pigs to cope with a challenging situation. Using PAP in the pig industry seems interesting to limit unwanted consequences of farm practices on animal welfare and productivity, by promoting their adaptation.

Impact of Semiochemicals Binding to Fel d 1 on Its 3D Conformation and Predicted B-Cell Epitopes Using Computational Approaches.

The major cat allergen Fel d 1 is a tetrameric glycoprotein from the secretoglobin superfamily. Fel d 1's biological role is unknown, but it has been previously shown that it participates in semiochemical binding/transportation. Fel d 1 has linear epitopes, but its conformational epitope sites remain unclear. In this study, we predicted the B-cell epitopes of Fel d 1 and explored semiochemical dynamics with epitopes using bioinformatics tools. The epitope residues were tabulated for chains 1 and 2 and the heterodimers of Fel d 1. The residual interactions of Fel d 1 with IgE were evaluated, and the prominent epitope sites were predicted. The molecular dynamics simulation (MDS) of Fel d 1 was performed with seven reported semiochemicals to evaluate the Fel d 1-ligand complex stability and decipher the semiochemical effect on Fel d 1 conformational epitopes. Fel d 1-lauric acid, Fel d 1-oleic acid, and Fel d 1-progesterone showed more stability and less fluctuation than other compounds. Fel d 1-linoleic acid and Fel d 1-pregnenolone displayed the most unstable complex with fluctuations. The effects of conformational changes on epitopes are discussed. All the ligand complexes drive substantial fluctuation towards the functionally exposed IgE-binding epitopes. Fel d 1 could be examined for its ligand-binding and conformational changes caused by mutations of B-cell epitopes.

Does the Environmental Air Impact the Condition of the Vomeronasal Organ? A Mouse Model for Intensive Farming.

Chemical communication in mammals is ensured by exchanging chemical signals through the vomeronasal organ (VNO) and its ability to detect pheromones. The alteration of this organ has been proven to impact animal life, participating in the onset of aggressive behaviors in social groups. To date, few studies have highlighted the possible causes leading to these alterations, and the farming environment has not been investigated, even though irritant substances such as ammonia are known to induce serious damage in the respiratory tract. The goal of this study was to investigate the environmental impact on the VNO structure. Thirty mice were split into three groups, one housed in normal laboratory conditions and the other two in confined environments, with or without the release of litter ammonia. VNOs were analyzed using histology and immunohistochemistry to evaluate the effect of different environments on their condition. Both restricted conditions induced VNO alterations (p = 0.0311), soft-tissue alteration (p = 0.0480), and nonsensory epithelium inflammation (p = 0.0024). There was glycogen accumulation (p < 0.0001), the olfactory marker protein was underexpressed (p < 0.0001), and Gαi2 positivity remained unchanged while Gαo expression was upregulated in confined conditions. VNO conditions seemed to worsen with ammonia, even if not always significantly. These murine model results suggest that the housing environment can strongly impact VNO conditions, providing novel insights for improving indoor farming systems.

Can a semiochemical deter cats from trash cans?

OBJECTIVES: The interaction between free-roaming cats and humans can lead to cohabitation issues. One such issue is the nuisance caused when cats scavenge and scatter garbage, particularly in cities. This study aimed to investigate the impact of a semiochemical on cat avoidance behaviour. METHODS: Five cats were included in the study, in a crossover design. A 10 min choice test involving two garbage cans, one sprayed with the semiochemical and the other sprayed with a control solution, was performed in a controlled setting. The parameters observed were exploration, proximity, latency to approach and first choice. RESULTS: Cats spent less time exploring (general linear mixed model [GLMM]; degrees of freedom [DF] = 1; F = 14.23; P <0.0001) and in proximity to the treated garbage can (GLMM; DF = 1; F = 13.25; P <0.0001). Additionally, although there was no significant difference in the first choice (P = 0.5300), the latency to approach tended to be longer for the treated garbage can (frailty model; DF = 1; χ2 = 3.49; P = 0.0610). CONCLUSIONS AND RELEVANCE: The present study demonstrated that this semiochemical can deter cats from trash cans in a controlled setting. Further studies are needed to explore the efficiency of the semiochemical deterrent in a real-world setting to improve human-cat cohabitation.

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.

Dental Pain in Cats: A Prospective 6-Month Study.

Dental pathology is among the most ubiquitous diseases in cats of all ages. Dental pain is yet to be fully understood in cats and therefore its presence is often missed. To better understand feline dental disease as a pain trigger during routine examination and whether disease severity correlates to the degree of pain, a 6-month prospective study in a cats' only veterinary hospital in Portugal was conducted. Sixty-four cats that randomly presented for different clinical procedures were evaluated. Dental and periodontal abnormalities (primary dental parameters, PDP), as well as clinical signs related to dental pain (secondary dental parameters, SDP), were assessed. All cats underwent an oral cavity examination, upon which, the Feline Acute Pain Scale from Colorado State University Veterinary Teaching Hospital (CPS), was used in order to assess pain. Six PDP (periodontal disease, gingival index, calculus index, tooth resorption, tooth fracture and missing teeth) and five SDP (mouth discomfort, halitosis, hypersalivation, difficulty in holding food and several attempts at prehension of food), were compared with CPS pain scores. All SDP were significantly associated to higher CPS pain scores (p < 0.05). The number of missing teeth was significantly associated to higher CPS pain scores (p < 0.0001). A trend was observed between higher CPS pain scores and tooth resorption (p = 0.08). This study concluded that cats with dental disease feel pain during clinical examination and the pain increases as the severity of the disease progresses.

Isolation and Characterization of Cat Olfactory Ecto-Mesenchymal Stem Cells.

The olfactory mucosa contains olfactory ecto-mesenchymal stem cells (OE-MSCs) which show stemness features, multipotency capabilities, and have a therapeutic potential. The OE-MSCs have already been collected and isolated from various mammals. The aim of this study was to evaluate the feasibility of collecting, purifying and amplifying OE-MSCs from the cat nasal cavity. Four cats were included in the study. Biopsies of olfactory mucosa were performed on anesthetized animals. Then, the olfactory OE-MSCs were isolated, and their stemness features as well as their mesodermal differentiation capabilities were characterized. Olfactory mucosa biopsies were successfully performed in all subjects. From these biopsies, cellular populations were rapidly generated, presenting various stemness features, such as a fibroblast-like morphology, nestin and MAP2 expression, and sphere and colony formation. These cells could differentiate into neural and mesodermal lineages. This report shows for the first time that the isolation of OE-MSCs from cat olfactory mucosa is possible. These cells showed stemness features and multilineage differentiation capabilities, indicating they may be a promising tool for autologous grafts and feline regenerative medicine.

Influencing Elimination Location in the Domestic Cat: A Semiochemical Approach.

In the domestic cat, elimination at an inappropriate location is considered by cat owners and non-cat owners as an undesirable behaviour. The aim of the study was to assess the effect of a semiochemical formulation, reconstituted volatile fraction of cat anal gland secretions on the elimination behaviour of domestic cats. The study was conducted in four catteries, which housed 33 cats, using 37 litter trays and followed a randomised crossover design using the litter tray as the experimental unit. The parameters studied included daily elimination (urine plus stools) weight, urine weight, stool weight, elimination type and urine/stool quantity scoring. The parameters were analysed using GLMM with SAS 9.4 software. Four out of the six parameters studied showed a treatment effect, consistently in favour of cats defecating significantly less in the litter trays sprayed with the treatment versus litter trays sprayed with the control (elimination weight p = 0.0199; elimination type p = 0.0251; stool weight p = 0.0005 and stool quantity p = 0.003). These results demonstrate that an intraspecific semiochemical message originating from cat anal glands influences cats' defecation location.

A Feline Semiochemical Composition Influences the Cat's Toileting Location Choice.

Unwanted toileting is amongst the most undesirable behaviors in domestic cats and can lead to conflicts between cats and the communities they are living in. This study aimed to confirm the effect of a semiochemical composition, reconstituted volatile fraction derived from cat anal glands, on the elimination behavior of domestic cats. A total of 31 cats were tested individually, for 23 h, in a blinded randomized choice test, with two litter trays, one sprayed with the treatment and the other with the control. Parameters included elimination weight, urine only weight, the record of the elimination type and counting of urine spots and stools, exploration duration of each litter tray, and first and second choice of litter tray to eliminate. Across all parameters, cats urinated and defecated significantly less in the litter tray where the semiochemical composition was sprayed than in the litter tray where the control was sprayed (for example: elimination weight p < 0.0001; urine only weight p < 0.0001; exploration duration p < 0.0001, and first elimination choice p < 0.0001). These results demonstrate that a semiochemical composition-derived from cat anal glands significantly decreases elimination at the location where it is sprayed. Future research is warranted to explore the possibility to manage unwanted toileting using this semiochemical composition.

Is There a Link between Vomeronasalitis and Aggression in Stable Social Groups of Female Pigs?

The vomeronasal organ (VNO) is a bilateral chemosensory structure strongly involved in animal behaviour, thanks to its sensory epithelium (VNSE) that detects pheromones. Experimental VNO lesions can impair social, reproductive and maternal behaviour, while feline spontaneous vomeronasalitis has been associated with aggression. This study aimed to describe vomeronasalitis in farm pigs and explore its association with intraspecific behavioural alterations. Using 38 six-month-old pigs, the skin lesion score based on Welfare Quality® protocols was obtained during the fattening period. The seventy-six VNOs from these pigs were stained in haematoxylin-eosin for histological examinations. VNSE inflammation was classified considering its intensity. Skin lesions data were compared to vomeronasalitis. There were 34% of pigs that showed unilateral VNSE inflammation, while 66% were bilaterally affected. The mean ± SD number of skin lesions/animal was 4.4 ± 2.82, and 34% of pigs scored 1 (moderately wounded animals) at least once during the fattening period. Statistical analysis showed an association between bilateral vomeronasalitis and skin lesion score (p < 0.05) and between bilateral moderate vomeronasalitis and skin lesions number (p < 0.01). This is the first report linking vomeronasalitis to social life in farm animals. Considering the role of social life in animal welfare, our data opens a research field linking pathology to animal behaviour.

Sex Differences in Mouse Exploratory Behaviour to Fel d 1, a Cat ABP-Like Protein.

Fel d 1 is a cat protein abundantly released and found in their habitat and is closely related to mouse androgen-binding proteins (ABPs). We hypothesized that mice have developed chemical communication mechanisms to detect and avoid this protein. We tested purified natural Fel d 1, a fox faeces molecule (TMT) as a positive control, and a negative control (purified water) in three different mouse groups (n = 14 each) to evaluate exploratory behaviour and stress responses. The mice did not show clear avoidance or stress responses to Fel d 1. Our results demonstrated a sex-treatment interaction for Fel d 1, with males spending more time in the areas treated with Fel d 1 than in the untreated areas (p = 0.018). This sex-treated area interaction was also not observed for either the blank or TMT. These results suggest that Fel d 1 from domestic cats could be recognized differently by male and female mice. These sex differences could be linked to the sexual role of ABP proteins and the ABP-like characteristics of Fel d 1.

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.

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.

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.

Histological and Immunohistochemical Characterization of Vomeronasal Organ Aging in Mice.

The vomeronasal organ (VNO) plays a crucial role in animal behavior since it is responsible for semiochemical detection and, thus, for intra- and interspecific chemical communication, through the vomeronasal sensory epithelium (VNSE), composed of bipolar sensory neurons. This study aimed to explore a well-recognized cause of neuronal degeneration, only rarely explored in this organ: aging. Murine VNOs were evaluated according to 3 age groups (3, 10, and 24 months) by histology to assess VNSE changes such as cellular degeneration or glycogen accumulation and by immunohistochemistry to explore nervous configuration, proliferation capability, and apoptosis with the expression of olfactory marker protein (OMP), Gαi2, Gαo, Ki-67, and cleaved caspase-3 proteins. These markers were quantified as percentages of positive signal in the VNSE and statistical analyses were performed. Cellular degeneration increased with age (p < 0.0001) as well as glycogen accumulation (p < 0.0001), Gαo expression (p < 0.0001), and the number of cleaved-caspase3 positive cells (p = 0.0425), while OMP and Gαi2 expressions decreased with age (p = 0.0436 and p < 0.0001, respectively). Ki67-positive cells were reduced, even if this difference was not statistically significant (p = 0.9105). Due to the crucial role of VNO in animal life, this study opens the door to interesting perspectives about chemical communication efficiency in aging animals.

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.

The provision of toys to pigs can improve the human-animal relationship.

It is now widely recognised that a positive human-animal relationship is beneficial not only for farm animals' welfare but also for productivity and the quality of products. A better understanding of animal emotions is an important goal in disciplines ranging from neuroscience to animal welfare science, but few reliable tools exist for measuring these emotions. In this study, whether the provision of toys to solicit play behaviour in pigs is associated with a change in the human-animal relationship and the emotional state of pigs was investigated. We involved a group of sixteen mini-pigs housed in an experimental setting and the use of a preliminary test called the 'strange person' test. After a Control and a Play session (with medium-sized dog toys, balls with ropes), the strange person test was performed. During the test, a person wearing a colourful overall, a hood, a mask, gloves and boots (unknown person with an odd appearance) entered the pen, where 2 mini-pigs were housed, for a 2-min video recording. The strange person test results after the Play and Control sessions were compared. The results showed that the latency to approach the person (duration in seconds) and the duration for which the pig was distant from the strange person (duration in seconds) were significantly lower after the Play session than after the Control session (Degrees of Freedom =30; Statistic of the F test =39.1; p < 0.0001 and Degrees of Freedom =15; Statistic of the F test =54.3; p < 0.0001, respectively). The duration of direct contact (duration in seconds) (Degrees of Freedom =15; Statistic of the F test =14.8; p = 0.002), the need to separate the pig from the strange person (frequency) (Degrees of Freedom =30; Statistic of the F test =9.3; p = 0.005) and the duration of tail movement (duration in seconds) (Degrees of Freedom =15; Statistic of the F test =12.6; p = 0.003) were all significantly higher after the Play sessions than after the Control sessions. Overall, the results suggest a change in the human-animal relationship after the Play sessions: the pigs seemed to be less fearful and more inclined to interact with the strange person, showing a more positive emotional state. This preliminary study suggests that the provision of toys, and more precisely, the opportunity to perform object play behaviour, and sometimes, spontaneously, social play behaviour, can improve the human-animal relationship. Additional research to explore this topic thoroughly may yield interesting results because a positive emotional state of the animals and a good human-animal relationship are essential to ensure good quality of life of farm animals.

Single fluff-spray application of mother hen uropygial secretion analogue positively influences bursa of Fabricius development and the heterophil-to-lymphocyte ratio in ROSS 308 chicks.

Stress is an important cause of illness and mortality in chick production. Stressors such as manipulation, absence of maternal care, transport, and housing can lead to welfare issues, immunodepression, and decreased productivity. The mother hen uropygial secretion analogue (MHUSA), a synthetic analog of a maternal semiochemical secretion, has been proven to protect chicks and broilers against stress, significantly reducing the heterophil-to-lymphocyte ratio. The aim of the present study was to test the effects of the MHUSA on chicks' stress when single-sprayed on their fluff at the age of 1 d. Two-hundred eighty ROSS 308 chicks were included in the study. At day 1, each chick received a spray of 200 µL of a 2% MHUSA aqueous solution (140 chicks) or the same amount of the excipient (control group, 140 chicks), and then chicks were housed in 2 separate rooms. To assess the persistence of the MHUSA after this single application, fluff was sampled from 10 chicks every day for 7 d and at day 13 and 19, weighed, placed in dichloromethane, and analyzed by gas chromatography. Blood smears and the bursa of Fabricius were collected every 3 d from 10 chicks of each group for 36 d to assess the heterophil-to-lymphocyte ratio and the bursa weight-to-BW ratio, respectively. Gas chromatography analysis showed that the MHUSA was present on chick fluff until day 5. The statistical analysis revealed that the heterophil-to-lymphocyte ratio was lower in the MHUSA group at day 4, 7, and 9 (P < 0.0001 for day 4 and 7; P = 0.0377 for day 9). The bursa weight-to-BW ratio was significantly higher in the MHUSA group than in the control group from day 4 until day 29. These results confirm the beneficial effects of the MHUSA on chicks' adaptation to the new environment and on bursa of Fabricius development, suggesting its potential role in improving chicks' immune response.