Genome-wide association study for morphological and hunting-behavior traits in Braque Français Type Pyrénées dogs: A preliminary study.

High-throughput genotyping offers great potential to increase our understanding of the genomic basis of canid variation. Braque Français Type Pyrénées (BRA) are smart, agile, and friendly dogs originally developed for tracking, hunting, and retrieving feathered game. On a population of 44 unrelated BRA dogs, single nucleotide polymorphism (SNP) genotype data from the CanineHD Whole-Genome Genotyping BeadChip and evaluation scores for 12 traits related to morphology and hunting performance were available. After quality filtering, 95,859 SNPs on the 38 dog autosomes (CFA) were retained. Phenotypic scores were expressed on a scale from 1 (worst) to 6 (best) and were mostly poorly to moderately correlated except for some morphological traits (e.g. r = 0.81 between the conformation of the head and that of the eye). From GWAS, a total of 378 SNP-phenotype associations with posterior odds of association > 1 have been detected. The strongest associations were found for the eye conformation, for the skull/muzzle ratio, and for connection to the hunter. These included both new and previously identified markers and genes potentially involved with type and behavior traits in BRA. Six of the significant markers mapped within SETDB2, a gene known to be related to pointing behavior in dogs. These results advance our understanding of the genetic basis for morphology and hunting behavior in dogs and identify new variants which are potential targets for further research.

Nuclear Magnetic Resonance (NMR) Metabolomics: Current Applications in Equine Health Assessment.

Metabolomics can allow for the comprehensive identification of metabolites within biological systems, at given time points, in physiological and pathological conditions. In the last few years, metabolomic analysis has gained popularity both in human and in veterinary medicine, showing great potential for novel applications in clinical activity. The aim of applying metabolomics in clinical practice is understanding the mechanisms underlying pathological conditions and the influence of certain stimuli (i.e., drugs, nutrition, exercise) on body systems, in the attempt of identifying biomarkers that can help in the diagnosis of diseases. Proton Nuclear Magnetic Resonance spectroscopy (1H-NMR) is well tailored to be used as an analytical platform for metabolites' detection at the base of metabolomics studies, due to minimal sample preparation and high reproducibility. In this mini-review article, the scientific production of NMR metabolomic applications to equine medicine is examined. The research works are very different in methodology and difficult to compare. Studies are mainly focused on exercise, reproduction, and nutrition, other than respiratory and musculoskeletal diseases. The available information on this topic is still scant, but a greater collection of data could allow researchers to define new reliable markers to be used in clinical practice for diagnostic and therapeutical purposes.

A Prospective, Blinded, Open-Label Clinical Trial to Assess the Ability of Fluorescent Light Energy to Enhance Wound Healing after Mastectomy in Female Dogs.

Mammary gland tumors represent the most frequently diagnosed malignant neoplasm in intact female dogs, and surgical removal represents the current gold standard treatment. To promote wound healing and prevent possible bacterial contamination, perioperative antimicrobials are commonly used in clinical practice, even though there are no publications establishing guidelines for the use of such drugs in canine mastectomy. The aim of the present study was to evaluate the ameliorative effect of fluorescent light energy on the quality of the healing process after mastectomy surgery in female dogs, in the absence of perioperative antimicrobial administration. Nine female dogs received a multiple-gland mastectomy due to gland tumors and received FLE application immediately after surgery and then five days after. The surgical incisions were evaluated by a blind investigator over time using the Modified Hollander Cosmesis and Modified Draize Wound Healing Score systems. Statistical analysis revealed a significant ameliorative effect of FLE in the control of step-off borders, contour irregularities, and excessive distortion. In addition, erythema, edema, and serous discharge were lower for those wounds managed with FLE. These results underscore the advantageous impact of FLE on the healing of post-mastectomy wounds in female dogs, offering the dual benefits of reducing potential infection risks and lessening the home care burden for pet owners.

Pro-Inflammatory Cytokines Enhanced In Vitro Cytotoxic Activity of Clostridioides difficile Toxin B in Enteric Glial Cells: The Achilles Heel of Clostridioides difficile Infection?

Bacterial infections are characterized by an inflammatory response, which is essential for infection containment but is also responsible for negative effects on the host. The pathogen itself may have evolved molecular mechanisms to antagonize the antimicrobial effects of an inflammatory response and to enhance its pathogenicity using inflammatory response mediators, such as cytokines. Clostridioides difficile (C. difficile) infection (CDI) causes gastrointestinal diseases with markedly increasing global incidence and mortality rates. The main C. difficile virulence factors, toxin A and B (TcdA/TcdB), cause cytopathic/cytotoxic effects and inflammation. We previously demonstrated that TcdB induces enteric glial cell (EGC) apoptosis, which is enhanced by the pro-inflammatory cytokine tumor necrosis factor alpha plus interferon gamma (CKs). However, it is unknown whether CKs-enhanced TcdB cytotoxicity (apoptosis/necrosis) is affected by the timing of the appearance of the CKs. Thus, we simulated in vitro, in our experimental model with TcdB and EGCs, three main situations of possible interactions between TcdB and the timing of CK stimulation: before TcdB infection, concomitantly with infection, or at different times after infection and persisting over time. In these experimental conditions, which all represent situations of possible interactions between C. difficile and the timing of CK stimulation, we evaluated apoptosis, necrosis, and cell cycle phases. The CKs, in all of these conditions, enhanced TcdB cytotoxicity, which from apoptosis became necrosis when CK stimulation persisted over time, and was most relevant after 48 h of TcdB:EGCs interaction. Particularly, the enhancement of apoptosis by CKs was dependent on the TcdB dose and in a less relevant manner on the CK stimulation time, while the enhancement of necrosis occurred always independently of the TcdB dose and CK stimulation time. However, since in all conditions stimulation with CKs strongly enhanced the TcdB cytotoxicity, it always had a negative impact on C. difficile pathogenicity. This study might have important implications for the treatment of CDI.

Exploring fluorescent light energy as management option for canine superficial bacterial folliculitis.

Superficial bacterial folliculitis (SBF) represents a common dermatological diagnosis in dogs that can be successfully managed with either topical and/or systemic treatments. In the present study we evaluated the efficacy of a fluorescent light energy (FLE) device as sole management for SBF. The same FLE device has been shown, as adjunct therapy to systemic antibiotic or alone, to effectively control clinical manifestation of interdigital furunculosis. Twenty dogs were randomized to receive FLE once (six dogs) or twice (six dogs) weekly in comparison with oral anti-biotic (eight dogs) until complete healing. FLE regimen was able to significantly reduce the time needed to clinical resolution for oral antibiotic, supporting owners' compliance and welfare of dogs.

Clostridioides difficile Toxin B Induced Senescence: A New Pathologic Player for Colorectal Cancer?

Clostridioides difficile (C. difficile) is responsible for a high percentage of gastrointestinal infections and its pathological activity is due to toxins A and B. C. difficile infection (CDI) is increasing worldwide due to the unstoppable spread of C. difficile in the anthropized environment and the progressive human colonization. The ability of C. difficile toxin B to induce senescent cells and the direct correlation between CDI, irritable bowel syndrome (IBS), and inflammatory bowel diseases (IBD) could cause an accumulation of senescent cells with important functional consequences. Furthermore, these senescent cells characterized by long survival could push pre-neoplastic cells originating in the colon towards the complete neoplastic transformation in colorectal cancer (CRC) by the senescence-associated secretory phenotype (SASP). Pre-neoplastic cells could appear as a result of various pro-carcinogenic events, among which, are infections with bacteria that produce genotoxins that generate cells with high genetic instability. Therefore, subjects who develop IBS and/or IBD after CDI should be monitored, especially if they then have further CDI relapses, waiting for the availability of senolytic and anti-SASP therapies to resolve the pro-carcinogenic risk due to accumulation of senescent cells after CDI followed by IBS and/or IBD.

Sensitivity and specificity of in vivo COVID-19 screening by detection dogs: Results of the C19-Screendog multicenter study.

Trained dogs can recognize the volatile organic compounds contained in biological samples of patients with COVID-19 infection. We assessed the sensitivity and specificity of in vivo SARS-CoV-2 screening by trained dogs. We recruited five dog-handler dyads. In the operant conditioning phase, the dogs were taught to distinguish between positive and negative sweat samples collected from volunteers' underarms in polymeric tubes. The conditioning was validated by tests involving 16 positive and 48 negative samples held or worn in such a way that the samples were invisible to the dog and handler. In the screening phase the dogs were led by their handlers to a drive-through facility for in vivo screening of volunteers who had just received a nasopharyngeal swab from nursing staff. Each volunteer who had already swabbed was subsequently tested by two dogs, whose responses were recorded as positive, negative, or inconclusive. The dogs' behavior was constantly monitored for attentiveness and wellbeing. All the dogs passed the conditioning phase, their responses showing a sensitivity of 83-100% and a specificity of 94-100%. The in vivo screening phase involved 1251 subjects, of whom 205 had a COVID-19 positive swab and two dogs per each subject to be screened. Screening sensitivity and specificity were respectively 91.6-97.6% and 96.3-100% when only one dog was involved, whereas combined screening by two dogs provided a higher sensitivity. Dog wellbeing was also analyzed: monitoring of stress and fatigue suggested that the screening activity did not adversely impact the dogs' wellbeing. This work, by screening a large number of subjects, strengthen recent findings that trained dogs can discriminate between COVID-19 infected and healthy human subjects and introduce two novel research aspects: i) assessement of signs of fatigue and stress in dogs during training and testing, and ii) combining screening by two dogs to improve detection sensitivity and specificity. Using some precautions to reduce the risk of infection and spillover, in vivo COVID-19 screening by a dog-handler dyad can be suitable to quickly screen large numbers of people: it is rapid, non-invasive and economical, since it does not involve actual sampling, lab resources or waste management, and is suitable to screen large numbers of people.

Fluorescent Light Energy in the Management of Multi Drug Resistant Canine Pyoderma: A Prospective Exploratory Study.

The increase in prevalence of staphylococcal antimicrobial resistance has been also associated with pyoderma in dogs, and prolonged antibiotic treatment, as often needed in severe cases of pyoderma, has been related to influencing possible development of multidrug resistance (MDR). Fluorescent light energy (FLE) has been indicated to improve pyoderma lesions as adjunct therapy to systemic antibiotics. In the present study, we evaluated the effect of FLE on clinical signs of MDR canine deep pyoderma (CDP) and interdigital furunculosis (CIF) when administered as solely management. Sixteen client-owned dogs affected by CIF (five dogs) and CDP (eleven dogs) were scored using a dedicated scoring system and received a single FLE applications twice weekly, until clinical resolution was achieved. Mean time to achieve complete resolution was 5.20 ± 3.56 weeks (median 3 weeks) for CIF cases and 4.18 ± 1.47 weeks (median 4 weeks) for CDP ones. FLE shows promise as an aid to managing clinical signs while reducing reliance on antibiotics for MDR CDP and CIF. In this study, FLE was responsible for the decrease in lesion scores and resolution of MDR pyoderma infection without any adjunct therapy, having a potential useful role to play in antibiotic stewardship programs, efficiently promoting complete clinical resolution of MDR lesions while optimizing the use of antibiotics.

Measuring Biochemical Variables and Serum Amyloid A (SAA) in Working Mules in Central Italy.

According to FAO reports, the global mule population counts about 9 million mules. This hybrid cross of a male donkey and a female horse is mainly used for draft purposes because they are thought to be strong and hardy animals. Most consider mules to be less susceptible to disease and fatigue compared to horses. Therefore, the aim of this study was to investigate the effects of fieldwork on biochemical variables and serum amyloid A in working mules. Blood samples were collected from 10 healthy, female, working mules before and after 8 h of fieldwork. According to statistical analysis, a significant influence (p < 0.05) of fieldwork was found on mules' electrolyte profile with increased levels of sodium, chloride, and calcium, as well as on blood urea nitrogen and creatinine. After a day of fieldwork, serum sodium, chloride, calcium, urea, and creatinine concentrations were increased, supporting decreases in body water and renal blood flow. However, without comparison to a group of mules that were not exercised yet maintained under similar ambient conditions, it is uncertain whether these changes can be attributed to exercise. Further, no change in SAA concentration was found after exercise, indicating that the work performed did not result in systemic inflammation.

Fluorescence Biomodulation for Canine Interdigital Furunculosis: Updates for Once-Weekly Schedule.

Interdigital furunculosis is a common multifactorial, inflammatory disease of the canine interdigital skin in which lesions commonly become secondarily infected. Fluorescence biomodulation (FBM) administered twice weekly has shown to effectively control clinical manifestation as adjunct therapy to systemic antibiotic. Since twice weekly regimen could be unaffordable for some pet owners, the aim of this study was to evaluate the effect of once weekly application of FBM in combination with systemic antibiotic on clinical manifestations of canine interdigital pyoderma, comparing the results to those present in literature. Twelve dogs diagnosed with interdigital pyoderma received antibiotic plus once weekly FBM application. Dogs were scored until complete healing based on global lesion score and neutrophil engulfing bacterial score. The results obtained demonstrated that once weekly application of FBM exerts the same beneficial effect on interdigital furunculosis healing as per twice weekly, indicating that once weekly regimen is well tolerated and is yielding similar results to twice weekly applications.

Clostridioides difficile toxin B alone and with pro-inflammatory cytokines induces apoptosis in enteric glial cells by activating three different signalling pathways mediated by caspases, calpains and cathepsin B.

Clostridioides difficile infection (CDI) causes nosocomial/antibiotic-associated gastrointestinal diseases with dramatically increasing global incidence and mortality rates. The main C. difficile virulence factors, toxins A and B (TcdA/TcdB), cause cytopathic/cytotoxic effects and inflammation. We demonstrated that TcdB induces caspase-dependent, mitochondria-independent enteric glial cell (EGC) apoptosis that is enhanced by the pro-inflammatory cytokines TNF-α and IFN-γ (CKs) by increasing caspase-3/7/9 and PARP activation. Because this cytotoxic synergism is important for CDI pathogenesis, we investigated the apoptotic pathways involved in TcdB- and TcdB + CK-induced apoptosis indepth. EGCs were pre-treated with the inhibitors BAF or Q-VD-OPh (pan-caspase), Z-DEVD-fmk (caspase-3/7), Z-IETD-fmk (caspase-8), PD150606 (calpains), and CA-074Me (cathepsin B) 1 h before TcdB exposure, while CKs were given 1.5 h after TcdB exposure, and assays were performed at 24 h. TcdB and TcdB + CKs induced apoptosis through three signalling pathways activated by calpains, caspases and cathepsins, which all are involved both in induction and execution apoptotic signalling under both conditions but to different degrees in TcdB and TcdB + CKs especially as regards to signal transduction mediated by these proteases towards downstream effects (apoptosis). Calpain activation by Ca2+ influx is the first pro-apoptotic event in TcdB- and TcdB + CK-induced EGC apoptosis and causes caspase-3, caspase-7 and PARP activation. PARP is also directly activated by calpains which are responsible of about 75% of apoptosis in TcdB and 62% in TcdB + CK which is both effector caspase-dependent and -independent. Initiator caspase-8 activation mediated by TcdB contributes to caspase-3/caspase-7 and PARP activation and is responsible of about 28% of apoptosis in both conditions. Caspase-3/caspase-7 activation is weakly responsible of apoptosis, indeed we found that it mediates 27% of apoptosis only in TcdB. Cathepsin B contributes to triggering pro-apoptotic signal and is responsible in both conditions of about 35% of apoptosis by a caspase-independent manner, and seems to regulate the caspase-3 and caspase-7 cleaved fragment levels, highlighting the complex interaction between these cysteine protease families activated during TcdB-induced apoptosis. Further a relevant difference between TcdB- and TcdB + CK-induced apoptosis is that TcdB-induced apoptosis increased slowly reaching at 72 h the value of 18.7%, while TcdB + CK-induced apoptosis increased strongly reaching at 72 h the value of 60.6%. Apoptotic signalling activation by TcdB + CKs is enriched by TNF-α-induced NF-κB signalling, inhibition of JNK activation and activation of AKT. In conclusion, the ability of C. difficile to activate three apoptotic pathways represents an important strategy to overcome resistance against its cytotoxic activity.

Invisible steps for a global endemy: molecular strategies adopted by Clostridioides difficile.

Clostridioides difficile infection (CDI) is on the rise worldwide and is associated with an increase in deaths and socio-health burden. C. difficile has become ubiquitous in anthropized environments because of the extreme resistance of its spores. Based on the epidemiological data and knowledge of molecular pathogenesis of C. difficile, it is possible to predict its progressive colonization of the human population for the following reasons: first, its global spread is unstoppable; second, the toxins (Tcds) produced by C. difficile, TcdA and TcdB, mainly cause cell death by apoptosis, but the surviving cells acquire a senescence state that favours persistence of C. difficile in the intestine; third, proinflammatory cytokines, tumour necrosis factor-α and interferon-γ, induced during CDI, enhance the cytotoxicity of Tcds and can increase the survival of senescent cells; fourth, Tcds block mobility and induce apoptosis in immune cells recruited at the infection site; and finally, after remission from primary infection or relapse, C. difficile causes functional abnormalities in the enteric glial cell (EGC) network that can result in irritable bowel syndrome, characterized by a latent inflammatory response that contributes to C. difficile survival and enhances the cytotoxic activity of low doses of TcdB, thus favouring further relapses. Since a 'global endemy' of C. difficile seems inevitable, it is necessary to develop an effective vaccine against Tcds for at-risk individuals, and to perform a prophylaxis/selective therapy with bacteriophages highly specific for C. difficile. We must be aware that CDI will become a global health problem in the forthcoming years, and we must be prepared to face this menace.

Faecal proteome in clinically healthy dogs and cats: Findings in pooled faeces from 10 cats and 10 dogs.

BACKGROUND: In the scientific literature, there are only a few manuscripts available on small animal faecal proteomics. METHODS: In the present pilot study, this evaluation was performed using pooled faecal samples from 10 clinically healthy dogs and, for the first time, in 10 clinically healthy cats by mean of two-dimensional electrophoresis followed by liquid chromatography-tandem mass spectrometry. RESULTS: Our results showed the presence of nine (albumin, alkaline phosphatase, chymotrypsin-C-like, cytosol aminopeptidase, elastase-3B/proteinase E, immunoglobulins and nuclear pore membrane glycoprotein 210) and 14 (albumin, caspase recruitment domain-containing protein, chymotrypsin-like, deleted in malignant brain tumours 1 protein-like, hypothetical protein LOC107375, immunoglobulin, kallikrein-1, superoxide dismutase, transthyretin precursor, interstitial collagenase-like) different proteins in canine and feline faeces, respectively. CONCLUSION: These preliminary findings document the presence of a range of proteins in the faeces of apparently healthy dogs and cats and may serve as a basis for larger, prospective studies to establish reference proteomic data against which diseased populations can be compared.

Updates on Laboratory Evaluation of Feline Cardiac Diseases.

Laboratory tests can be altered in cardiovascular diseases and the investigation of specific tests or biomarkers may provide additional information about myocardial damage. Traditional laboratory tests, such as cell blood count, serum biochemistry, and coagulation, can be useful in investigating patients, but are not specific. However, markers like Troponin and Natriuretic Peptides may possibly furnish further data on myocardium damage and can be used in both studying and monitoring cats with cardiac disease. Moreover, the evaluation of the thyroid profile is very important as hyperthyroid cats concomitant cardiovascular diseases are very common and they can also be a direct consequence of endocrinopathy. The purpose of this manuscript is to provide the widest possible overview of what is present in the literature about the feline clinical pathology of heart diseases through a rational division of the main alterations of traditional tests and biomarkers.

Increased Expression of Toll-Like Receptor 4 in Skin of Dogs with Discoid Lupus Erythematous (DLE).

Discoid lupus erythematous (DLE) is a common autoimmune skin disorder of dogs where keratinocytes play a pivotal role both in the innate and adaptive immune responses. As for the innate response, pattern recognition receptors (PRR), including Toll-like receptors (TLRs), can activate macrophages and immune tissue cells allowing for transmission and transduction of signals through cytokine and chemokine release to improve host defenses. In particular, TLR4 can also recognize endogenous molecules such as heat shock proteins produced during reactions to tissue damage. The aim of this study was to evaluate the expression of TLR4, a bacterial lipopolysaccharide sensor, in the skin of dogs with DLE and in normal skin to evaluate a possible involvement of this receptor in the disease pathogenesis. Skin samples of affected dogs had a diffuse and intense expression of TLR4 in the epidermis. Also, the inflammatory infiltrates were immunolabelled. The expression was significantly higher in DLE skin compared to normal skin (**** p < 0.0001). In conclusion, dogs with DLE showed an altered expression of TLR4, which might play an important pathogenic role in the ongoing immunopathologic process, thus being considered a valuable therapeutic potential target for DLE.

Use of nutraceuticals in the stallion: Effects on semen quality and preservation.

Nutritional supplements are widely used in the equine industry with the aim of improving horse health, sports or reproductive performances. Over the years, a number of studies have focused on investigating the effects of several dietary compounds on the quality and preservation of stallion semen. This paper reviews the literature available on the use of nutritional supplementation for the improvement of reproductive performance and semen quality in equine species, critically appraising the benefits and negative effects of several compounds found in complementary feeds such as PUFAs from different sources, vitamins and antioxidants, carnitine and botanical extracts. Different nutraceuticals have been highlighted to improve stallion fertility by providing optimal levels of antioxidants, with the most promising results obtained by the combination of PUFAs and antioxidants that resulted to be essential for the maintenance of normal reproductive functions and the reduction of cryodamage in cooled and frozen equine semen.

The Effectiveness of Fluorescent Light Energy as Adjunct Therapy in Canine Deep Pyoderma: A Randomized Clinical Trial.

A single centre, single-blinded, prospective, randomized, controlled clinical study was conducted to evaluate the effectiveness of twice weekly fluorescent light energy therapy (Phovia™) as adjunct to systemic antibiotics in the management of deep pyoderma in dogs. Dogs with clinical lesions consistent with deep pyoderma, positive bacterial culture, and showing neutrophil engulfing bacteria at cytology were included in the study. Assessments were undertaken weekly for 8 weeks and every 2 weeks thereafter until 12 weeks after enrolment. At each visit, lesions were scored and cytology was conducted to determine a neutrophil engulfing bacteria score. All dogs (Groups A and B) were treated with systemic antibiotic twice daily, and Group B received additionally Phovia twice weekly. Median treatment duration was 11.7 weeks for Group A and 5.7 weeks for Group B. After 8 weeks of treatment, the percentage of dogs that achieved clinical resolution was 35.0% and 88.0% for Groups A and B, respectively. Lesion scores showed highly statistically significant difference in favour of Group B from week 3 to 8, and neutrophil engulfing bacteria scores showed statistical difference from week 2 onwards in favour of Group B. These results indicate that Phovia, when used as an adjunct to systemic antibiotics, can accelerate time to clinical resolution in cases of canine deep pyoderma.

Current Applications and Future Perspectives of Fluorescence Light Energy Biomodulation in Veterinary Medicine.

The purpose of this review is to determine the state of the art of the mode of action and potential applications of fluorescence photobiomodulation in veterinary medicine. After a summary of the assets that have led the translation of such light-based therapies from bench side into clinical use, recent advances in canine dermatology using this brand-new approach are presented, and future scenarios where this type of care may provide benefits over the current standard care are highlighted.

Management of canine perianal fistula with fluorescence light energy: preliminary findings.

BACKGROUND: Canine perianal fistula (CPF) is a severe, painful, debilitating skin condition affecting the perianal skin. It often interferes with the quality of life of both dog and owner. Conventional medical treatment involves the use of immunosuppressive therapy; however, the successful resolution of lesions can be limited by poor owner compliance, adverse drug effects and dependence on costly therapies. HYPOTHESIS/OBJECTIVES: The present study aimed to assess the potential benefits of fluorescent light energy (FLE) on cases of CPF. ANIMALS: Four dogs with active perianal fistulas METHODS AND MATERIALS: FLE was applied as sole management therapy once a week with two consecutive applications in the same session for each dog until clinical signs had significantly improved, with weekly assessments for a six week period. Dogs were assessed by measuring the size of lesions at the start of the study and then weekly for six weeks, using planimetry software. Owners recorded vocalization and straining frequency scores during their pet's defaecation, and perianal licking frequency on a 0-5 point scale to assess the response to therapy. RESULTS: All dogs improved with FLE, achieving a significant reduction in vocalization, straining and licking after two weeks (P = 0.002). After five weeks of FLE therapy, lesional areas had significantly decreased (P = 0.04). Only one dog required more than seven applications. No adverse events were reported. CONCLUSION AND CLINICAL IMPORTANCE: FLE may be a promising alternative therapy for CPF.