Profile of gamma-delta (γδ) T lymphocytes in the peripheral blood of crossbreed dogs during stages of life and implication in aging

Abstract Background: Data on gamma-delta (γδ) T lymphocytes in the peripheral blood of dogs are scant, related only to healthy pure breed dogs and limited to a restricted age range. The aim of the study was to investigate the modulation of the γδ T lymphocyte (TCRγδ+) subpopulation in peripheral blood of crossbreed healthy dogs according to five identified stages of life: Puppy, Junior, Adult, Mature, Senior and to determine its implication in aging. A rigorous method of recruitment was used to minimize the influence of internal or external pressure on the immune response. Twenty-three intact female and twenty-four intact male dogs were enrolled. Blood samples were collected and immunophenotyping of peripheral blood T lymphocytes and γδ T cell subpopulations was performed. Results: The percentage of γδ T cells in peripheral blood lymphocytes was comparable with the value of 2.5% published by Faldyna and co-workers (2001), despite the percentage reported was investigated in less arranged age range groups and coming from four different dog pure breeds, whereas our data were recorded on wider age range groups and coming from crossbreed dogs. Therefore, the γδ T cell percentage (2.5%) is consistent and points out that such value is breed-independent. Statistical analysis highlighted differences in both percentage and absolute γδ T cells according to the stage of life. γδ T cells decreased significantly in the peripheral blood of elder dogs (Senior group) in comparison with previous stages of life (Puppy, Junior, and Adult groups). Differences in γδ T cells are significant and they are reported, for the first time, related to dog aging. Conclusions: The study confirms dogs to be among the animals with a low TCRγδ+ cell profile. A decrease of the TCRγδ+ subpopulation percentage was observed in elder dogs. TCRγδ+ cells of group S were different from those of groups P, J, and A. The differences are reported for the first time in dog aging. Identifying the stage of life when the decrease of γδ T lymphocytes starts can be useful for providing a rationale for drafting a wellness plan trial to support thymus immune functions and mitigate its functional exhaustion

Study of animal remains dug out during the excavations of a Nuragic village in Sardinia

This paper presents the results of the zooarchaeological analysis of the faunal materials brought to light during the excavations set up in the Nuragic village surrounding the Santu Antine Nuraghe, near Torralba (Sassari), Sardinia. Precisely, the remains come from the structure of the village named by archaeologists hut 12. They are 779 specimens out of thousands animal remains from the whole archaeological site. The majority of the rests belong to sheep (Ovis aries) or goats (Capra hircus), cattle (Bos taurus), pigs (Sus scrofa) and deers (Cervus elaphus). Such material may provide suggestions about the productive use of animals in the village and point out the importance of the economical management of animals in the Nuragic society. Indeed, many remains show signs with human origin, which testify that the hut may have been a workplace where bone fragments were processed to obtain different kind of tools

Influence of genetic selection on the myofibre type composition of porcine biceps femoris muscle: a comparative study of a purebred (Nero di Parma) and commercial hybrid pigs (Large White × Landrace × Duroc)

This study is aimed to evaluate histological features related to the different quality of meat cuts obtained from different pig breeds. We compared animals genetically selected to restore the original local purebred “Nero di Parma” pig, and highly selected fast-growing commercial hybrids. As weight is the main factor determining the time of slaughter, we compared equally weighing animals, sampled from slaughter for edible use, regardless of their age and sex, and immunohistochemically demonstrated the myofibre type composition of their biceps femoris muscle. In both groups we observed type I myofibres, situated as central islets, encircled by type IIA, IIX and IIB myofibres ordered in concentric rings according to the dynamic of their differentiation/maturation. However, the purebred pig muscles contained a smaller quantity of myofibres expressing the MyHC-IIb isoform, related to rapid postmortem glycolytic rate, meat toughness and poorer quality, in comparison to commercial hybrids. This proves that the latter are subjected to a more rapid transition in the expression of the different MyHC, probably as a consequence of genetic selection and breeding conditions, such as different feeding and housing reducing the possibility of continuous physical exercise. Further studies on the postnatal transitions timing of myosin heavy chain isoforms in functionally different muscles of various breeds are necessary to verify if they might be “artificially modulated”, with the aim to design breeding programs allowing a good balance between growth performance, muscularity and meat quality

Enterobacteriaceae and Salmonella contamination of wild boar (Sus scrofa) carcasses: comparison between different sampling strategies

During 2020, a total of 64 wild boar carcasses were tested for Enterobacteriaceae count (EBC), Salmonella and Yersinia enterocolitica in the abdominal region (i) within 5 h after hunting in the game collection point and (ii) before dressing and processing in the game-handling establishment (GHE) (49 carcasses—average time interval between (i) and (ii): 4.3 days). Because of COVID-19 restrictions, 15 carcasses were transported to a near slaughterhouse (average time interval between (i) and (ii): 2.3 days). Mesenteric lymph nodes (MLNs) were collected and tested for Salmonella and Y. enterocolitica. Results are shown in relation to sampling A (49 carcasses—GHE) and sampling B (15 carcasses—slaughterhouse). Sampling A: EBC median values were (i) 2.51 log10 CFU/cm2 and (ii) 2.79 log10 CFU/cm2. EBC increase between (i) and (ii) was statistically significant (p = 0.001). Salmonella prevalence on carcasses varied from (i) 2.0 to (ii) 6.1%. Sampling B: EBC median values were (i) 3.1 log10 CFU/cm2 and (ii) 3.32 log10 CFU/cm2. EBC increase between (i) and (ii) was not statistically significant (p = 0.191). Salmonella prevalence on carcasses varied from (i) 6.7 to (ii) 0.0%. The prevalence (sampling A + B) of lymphatic Salmonella carriers was 7.8% (5/64). From carcasses and/or MNLs, the serovars Enteritidis, Typhimurium, Agama, Zaiman and Diarizonae O:50 (z) were detected. Y. enterocolitica was never isolated. Long chilling periods prior to wild game processing should be avoided, and carcasses should be tested at GHE rather than after shooting to proper reflect the microbial load of wild boar meat entering the food chain

Chitosan-based scaffold modified with D-(+) raffinose for cartilage repair: an in vivo study

BackgroundOsteochondral defects significantly affect patients¿ quality of life and represent challenging tissue lesions, because of the poor regenerative capacity of cartilage. Tissue engineering has long sought to promote cartilage repair, by employing artificial scaffolds to enhance cell capacity to deposit new cartilage. An ideal biomaterial should closely mimic the natural environment of the tissue, to promote scaffold colonization, cell differentiation and the maintenance of a differentiated cellular phenotype. The present study evaluated chitosan scaffolds enriched with D-(+) raffinose in osteochondral defects in rabbits. Cartilage defects were created in distal femurs, both on the condyle and on the trochlea, and were left untreated or received a chitosan scaffold. The animals were sacrificed after 2 or 4 weeks, and samples were analysed microscopically.ResultsThe retrieved implants were surrounded by a fibrous capsule and contained a noticeable inflammatory infiltrate. No hyaline cartilage was formed in the defects. Although defect closure reached approximately 100% in the control group after 4 weeks, defects did not completely heal when filled with chitosan. In these samples, the lesion contained granulation tissue at 2 weeks, which was then replaced by fibrous connective tissue by week 4. Noteworthy, chitosan never appeared to be integrated in the surrounding cartilage.ConclusionsIn conclusion, the present study highlights the limits of D-(+) raffinose-enriched chitosan for cartilage regeneration and offers useful information for further development of this material for tissue repair

Immunophenotypical characterization of canine mesenchymal stem cells from perivisceral and subcutaneous adipose tissue by a species-specific panel of antibodies

Immunophenotypical characterization of mesenchymal stem cells is fundamental for the design and execution of sound experimental and clinical studies. The scarce availability of species-specific antibodies for canine antigens has hampered the immunophenotypical characterization of canine mesenchymal stem cells (MSC). The aim of this study was to select a panel of species-specific direct antibodies readily useful for canine mesenchymal stem cells characterization. They were isolated from perivisceral and subcutaneous adipose tissue samples collected during regular surgeries from 8 dogs. Single color flow cytometric analysis of mesenchymal stem cells (P3) deriving from subcutaneous and perivisceral adipose tissue with a panel of 7 direct anti-canine antibodies revealed two largely homogenous cell populations with a similar pattern: CD29+, CD44+, CD73+, CD90+, CD34−, CD45− and MHC-II− with no statistically significant differences among them. Antibody reactivity was demonstrated on canine peripheral blood mononuclear cells. The similarities are reinforced by their in vitro cell morphology, trilineage differentiation ability and RT-PCR analysis (CD90+, CD73+, CD105+, CD44+, CD13+, CD29+, Oct-4+ gene and CD31− and CD45− expression). Our results report for the first time a comparison between the immunophenotypic profile of canine MSC deriving from perivisceral and subcutaneous adipose tissue. The substantial equivalence between the two populations has practical implication on clinical applications, giving the opportunity to choose the source depending on the patient needs. The results contribute to routine characterization of MSC populations grown in vitro, a mandatory process for the definition of solid and reproducible laboratory and therapeutic procedures