Research Institute in Semiochemistry and Applied Ethology
Recent publications
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.
The union or reunion of animals with social groups can be a challenging situation, and little has been published about it when solitary species are concerned. Therefore, the aim of the present study was two-fold: (1) to advocate the need for systematic publications about strategies and the outcomes of reunion episodes in zoos and other facilities; and (2) to describe the behaviour of European wildcats (Felis silvestris silvestris) during one such episode, in which a female cat was reintroduced into her family social group using a gradual reunion procedure and cat appeasing pheromone (CAP) (spot-on). The study comprised three periods: the pre-reunion period (10 days, 20 sessions per wildcat), the post-reunion period (A, 5 days, 10 sessions per wildcat; B, 5 days, 10 sessions per wildcat) and the late-reunion period (5 days, 10 sessions per wildcat). In the post-reunion periods, all wildcats were together in the enclosure and were spotted with CAP pheromone. Per period, we collected data on individual and social behaviours. Individual behaviours, such as attention and self-grooming, were performed more in the pre-reunion than in the post-reunion period. Regarding social behaviours, we found that agonistic behaviours were performed more in the pre-reunion than in the post-reunion period. We observed behavioural changes over the course of the study, with behavioural patterns of the late-reunion periods resembling those of the pre-reunion period.
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.
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.
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.
The dams of gregarious animals must develop a close bond with their newborns to provide them with maternal care, including protection against predators, immunological transference, and nutrition. Even though lactation demands high energy expenditures, behaviors known as allo-nursing (the nursing of non-descendant infants) and allosuckling (suckling from any female other than the mother) have been reported in various species of wild or domestic, and terrestrial or aquatic animals. These behaviors seem to be elements of a multifactorial strategy, since reports suggest that they depend on the following: species, living conditions, social stability, and kinship relations, among other group factors. Despite their potential benefits, allonursing and allosuckling can place the health and welfare of both non-filial dams and alien offspring at risk, as it augments the probability of pathogen transmission. This review aims to analyze the biological and physi-ological foundations and bioenergetic costs of these behaviors, analyzing the individual and collective advantages and disadvantages for the dams’ own offspring(s) and alien neonate(s). We also include information on the animal species in which these behaviors occur and their impli-cations on animal welfare.
Doxorubicin is an extensively prescribed antineoplastic agent. It is also known for adverse effects, among which cardiotoxicity tops the list. The possible mechanism underlying doxorubicin (DOX)-mediated cardiotoxicity has been investigated in this study. Further, to reduce the DOX-mediated cardiotoxicity, DOX was conjugated with Chitosan Nanoparticles (DCNPs) and supplemented with propionic acid. Initially, the drug loading efficacy and conjugation of DOX with chitosan was confirmed by UV–Visible Spectroscopy (UV) and Fourier Transform Infrared Spectroscopy (FTIR). The average sizes of the synthesized Chitosan Nanoparticles (CNPs) and DCNPs were measured by Dynamic Light Scattering (DLS) analysis as 187.9 ± 1.05 nm and 277.3 ± 8.15 nm, respectively, and the zeta potential values were recorded as 55.2 ± 0.7 mV and 51.9 ± 1.0 mV, respectively. The size and shape of CNPs and DCNPs were recorded using a High-Resolution Electron Microscopy (HRTEM). The particles measured <30 nm and 33–84 nm, respectively. The toxic effects of DCNPs and propionic acid were evaluated in rat model. The data from the electrocardiogram (ECG), cardiac biomarkers, Peroxisome proliferator-activated receptor gamma (PPARγ) and histological observations indicated evidence of DOX-mediated cardiotoxicity, whereas the administration of DCNPs, as well as Propionic Acid (PA), brought about a restoration to normalcy and offered protection in the context of DOX-induced cardiotoxicity.
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.
Specific mate communication and recognition underlies reproduction and hence speciation. Our study provides new insights in Drosophila melanogaster premating olfactory communication. Mate communication evolves during adaptation to ecological niches and makes use of social signals and habitat cues. Female-produced, species-specific volatile pheromone (Z)-4-undecenal (Z4-11Al) and male pheromone (Z)-11-octadecenyl acetate (cVA) interact with food odour in a sex-specific manner. Furthermore, Z4-11Al, which mediates upwind flight attraction in both sexes, also elicits courtship in experienced males. Two isoforms of the olfactory receptor Or69a are co-expressed in the same olfactory sensory neurons. Z4-11Al is perceived via Or69aB, while the food odorant (R)-linalool is a main ligand for the other variant, Or69aA. However, only Z4-11Al mediates courtship in experienced males, not (R)-linalool. Behavioural discrimination is reflected by calcium imaging of the antennal lobe, showing distinct glomerular activation patterns by these two compounds. Male sex pheromone cVA is known to affect male and female courtship at close range, but does not elicit upwind flight attraction as a single compound, in to contrast to Z4-11Al. A blend of the food odour vinegar and cVA attracted females, while a blend of vinegar and female pheromone Z4-11Al attracted males, instead. Sex-specific upwind flight attraction to blends of food volatiles and male and female pheromone, respectively, adds a new element to Drosophila olfactory premating communication and is an unambiguous paradigm for identifying the behaviourally active components, towards a more complete concept of food-pheromone odour objects.
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
Objectives The cellulase enzyme is used for various industrial applications such as textile, paper, food and biofuel industries. Industrially, the fungal strain, Aspergillus niger, is widely used to produce cellulase enzymes. Emerging evidence has indicated the possible role of magnetic nanocomposites in enhancing cellulase production by A. niger. The cellulase/ iron oxide nanocomposites are already reported to be an eco-friendly method for modulating microbial biochemical characteristics. Methods The present study was used to assess the efficacy of cellulase/iron oxide magnetic composites (CMNPS) for the enhanced production of cellulase enzyme. The fungal strain Aspergillus niger was isolated from the soil samples by using standard techniques. The fungi were then transfected with CMNPS and further evaluated for the production of cellulase enzyme. Results Synthesis of the cellulase/iron oxide magnetic composites was characterized by UV, FTIR, and XRD. The produced CMNPS was used as substrate and enzyme production. The cellulase enzyme production by Aspergillus niger was analyzed by CMC (0.82 IU/ml enzyme activity) and FPA assay (0.039 IU/ml enzyme activity). CMNPS was also analyzed by CMC (0.74 IU/ml enzyme activity) and FPA assay (0.039 IU/ml enzyme activity). Reuse of CMNPS is the ultimate application and ecofriendly approach for the synthesis of cellulase enzymes. Conclusion The study concludes that the cellulase/ iron oxide nanocomposites may be a useful tool for the enhanced production of cellulase enzyme from the soil fungus A. niger.
Animals’ facial expressions are involuntary responses that serve to communicate the emotions that individuals feel. Due to their close co-existence with humans, broad attention has been given to identifying these expressions in certain species, especially dogs. This review aims to analyze and discuss the advances in identifying the facial expressions of domestic dogs and their clinical utility in recognizing pain as a method to improve daily practice and, in an accessible and effective way, assess the health outcome of dogs. This study focuses on aspects related to the anatomy and physiology of facial expressions in dogs, their emotions, and evaluations of their eyebrows, eyes, lips, and ear positions as changes that reflect pain or nociception. In this regard, research has found that dogs have anatomical configurations that allow them to generate changes in their expressions that similar canids—wolves, for example—cannot produce. Additionally, dogs can perceive emotions similar to those of their human tutors due to close human-animal interaction. This phenomenon—called “emotional contagion”—is triggered precisely by the dog’s capacity to identify their owners’ gestures and then react by emitting responses with either similar or opposed expressions that correspond to positive or negative stimuli, respectively. In conclusion, facial expressions are essential to maintaining social interaction between dogs and other species, as in their bond with humans. Moreover, this provides valuable information on emotions and the perception of pain, so in dogs, they can serve as valuable elements for recognizing and evaluating pain in clinical settings.
In some guide dog organisations, future guide dogs for blind individuals are required to undergo separation from their foster family from Monday to Friday as part of their training. These separations and the constantly changing environment may induce stress, thus impacting the welfare of these dogs and their performance. The aim of this study was therefore to evaluate this stress through physiological and behavioural measures. The results showed a significant increase in salivary cortisol levels at the time of separation (GLMM; DF = 2; F = 10.31; p < 0.0001). Additionally, the dogs were more passive on Friday than on other days (GLMM; DF = 2; F = 7.53; p = 0.0090), and “head movements” were expressed less frequently on Fridays (GLMM; DF = 2; F = 5.12; p = 0.0141). Performance increased across weeks, despite a lower “focused” score on Mondays (GLMM; DF = 2; F = 4.39; p = 0.0243). These results showed both adaptation to life in the kennel and that the dogs need to readapt to the situation each week. The increase in serotonin levels during the 3 weeks of testing (GLMM; DF = 2; F = 4.87; p = 0.0224) may also indicate that the dogs can adapt to the kennel environment. Therefore, this study questions the relevance of noncontinuous training programmes. In future research, it would be interesting to compare these results with those of a group of dogs staying at school on weekends.
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.
Subtilosin A, a cyclic peptide from Bacillus subtilis is known for its antimicrobial activity against a diverse range of bacteria. Herein, we report the specific interaction between subtilosin A against virulent proteins of Aeromonas hydrophila through in silico analysis. Aeromonas toxic proteins such as aerolysin and hemolysin were selected from the non-redundant database. The hemolysin protein was designed by homology modelling tool, and it was validated using Ramachandran plot. Then subtilosin A and target toxin proteins were energy minimized for further docking study. The whole docking experiments were done using antibody mode in Cluspro. Subtilosin A building an active interaction with Aeromonas toxins through H-bonds and protein–protein docking analysis revealed that the hemolysin has 6 H-bond interaction towards the antimicrobial target protein subtilosin A than aerolysin, which has 9 H-bonds. The most favourable interacting residues of subtilosin A are Thr6, Cys13, Ile19, Pro20, Asp21, Phe22, Glu23 and Gly35 involving in the strong H-bond formation and proceeds to inhibition of toxin. Hence, the study confirmed that the subtilosin A has more antimicrobial activity to inhibit the Aeromonas toxins by interacting with their binding site residues for preventing extracellular cleavage.
Angiotensin-converting enzyme (ACE) enhances blood pressure by making potential vasoconstrictor, Angiotensin-II from Angiotensin-I. Thus, it is the key target enzyme in cardiovascular disease therapy. Hence, in the present study an attempt has been made for identification and characterization of ACE inhibitory peptide from buffalo milk casein. Buffalo milk casein was isolated and digested with different digestive enzymes (pepsin, trypsin, chymotrypsin and their combination). Pepsin-Trypsin hydrolysates shown highest (72.55 ± 2.23%/50 μg) ACE inhibitory activity, in further ultra-filtration, < 1 kDa fraction shown the highest inhibitory activity among the other fractions. Below 1 kDa filtrate was further fractionated through RP-HPLC, fraction 11 exhibited the highest ACE inhibitory activity. Out of 15 peptides of 11th fraction (LC–MS/MS sequence) VLPVPQK is the novel peptide, which shown significant ACEI activity. Therefore, this study supports the notion that milk is a very good nutraceutical, and the novel peptide sequence can be utilized in commercial peptide synthesis for making functional food preparation.
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), G�i2, G�o, 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), G�o expression (p < 0.0001), and the number of cleaved-caspase3 positive cells (p = 0.0425), while OMP and G�i2 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.
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36 members
Sébastien Lebreton
  • Chemical Ecology
Alessandro Cozzi
  • Ethology and Neurosciences
Pietro Asproni
  • Department of Cellular Processes and Sustainable Interactions
Manuel Mengoli
  • Ethology and Neurosciences
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Patrick PAGEAT
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