Veterinary Sciences — Open Access Journal

The presence of BRAF variant V595E, as well as an increased cyclooxygenase-2 (COX-2) expression in canine transitional cell carcinoma (TCC) are well-described in the literature. The aim of the present study was to investigate the correlation between breed (terrier versus non-terrier dogs), histological grade, COX-2 expression, and BRAF mutation in canine TCC. Therefore, transmural TCC biopsies from 65 dogs (15 terriers, 50 non-terriers) were graded histologically into low- and high-grade. Immunohistochemical evaluation of the intensity of COX-2 expression was performed using an immunoreactive score (IRS). Exon 15 of chromosome 16 was examined for the BRAF variant c.1799T>A by TaqMan^® SNP assay. TCC was low-grade in 20 cases (one terrier, 19 non-terriers) and high-grade in 45 cases (14 terriers, 31 non-terriers). Contrary to humans, histological grade was not significantly correlated to the intensity of COX-2 expression. BRAF mutation was detected in 11/15 (73%) TCC of terriers and in 18/50 (36%) TCC of non-terriers. Histological grade and BRAF mutation were not correlated significantly (p = 0.2912). Terriers had a considerably higher prevalence of high-grade tumors (p < 0.0001), as well as of BRAF mutation (p ≤ 0.05) compared to non-terriers. In non-terriers, neoplasms with BRAF mutation showed a significantly higher intensity of COX-2 expression than those without BRAF mutation (p ≤ 0.05). In conclusion, in contrast to humans, testing for BRAF mutation in canine TCC is a sensitive diagnostic method especially in terriers (73%) and may be recommended as a screening test. However, evidence of BRAF mutation in canine TCC is not a predictor for the histological grade. Moreover, a positive correlation between histological grade and the intensity of COX-2 expression was not found. Further studies are necessary to clarify the clinical and prognostic relevance of the elevated intensity of COX-2 expression of TCC with BRAF mutation detected in non-terriers. Full article

The present study was conducted to evaluate the toxicity induced by the increasing doses of potassium dichromate in rabbit doe. Twenty-eight adult does of 6 months of age were divided into four groups (A, B, C, and D; n = 7), with comparable average body weight (bw). Group A rabbits received only distilled water daily and served as a control, while groups B, C, and D received, respectively, 10 mg/kg bw, 20 mg/ kg bw, and 40 mg/kg bw of potassium dichromate via gavage for 28 days, after which animals were anesthetized with ether vapor and sacrificed. Blood samples were obtained via cardiac puncture and collected without anticoagulant for biochemical dosages and with anticoagulant (EDTA) for complete blood count. Follicle stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) were dosed in serum and in homogenates of ovary with the help of AccuDiag^TM ELISA kits from OMEGA DIAGNOSTICS LTD (Scotland, England) while respecting the immuno-enzymatic method. Activities of superoxide dismutase (SOD), glutathione (GSH), catalase (CAT), and concentration of malondialdehyde (MDA) in liver, kidney, ovary and uterus were measured. Hematology revealed a significant (p < 0.05) decrease in mean values of hemoglobin and platelets while white blood cells and lymphocytes showed a significant (p < 0.05) increase in exposed groups. No significant (p > 0.05) difference was registered in monocytes, red blood cells, hematocrits, and plaquetocrits values with respect to the control. No matter the organ considered, no significant (p > 0.05) change was recorded in weight and volume. Nephrotoxicity analysis registered a significant (p < 0.05) increase in urea and creatinine, unlike renal tissue protein, which decreased significantly (p < 0.05). However, hepatotoxicity registered no significant (p > 0.05) variation in aspartate aminotransferase but total protein, alanine aminotransferase, and total cholesterol increased significantly (p < 0.05), while hepatic tissue protein revealed a significant (p < 0.05) decrease. Analysis on reproductive parameters showed a significant (p < 0.05) decrease in ovarian and uterine tissue proteins, as well as in follicle stimulating hormone, luteinizing hormone, and estradiol. Oxidative stress markers recorded no significant (p > 0.05) difference in glutathione reductase except in ovary where a significant (p < 0.05) decrease was seen when compared with the control, while catalase revealed a significant (p < 0.05) decrease, except in liver where there was no significant (p > 0.05) change. Superoxide dismutase and malondialdehyde recorded a significant (p < 0.05) decrease and increase respectively, with respect to the control. Results obtained from this study showed that the reduction process of chromium in tissues may cause the generation of reactive oxygen species, which are involved in hematoxic, nephrotoxic, hepatotoxic, and reproductive toxicity effects. Full article

Nematode parasites have a profound impact on humankind, infecting nearly one-quarter of the world’s population, as well as livestock. There is a pressing need for discovering nematicides due to the spread of resistance to currently used drugs. The free-living nematode Caenorhabditis elegans is a formidable experimentally tractable model organism that offers key advantages in accelerating nematicide discovery. We report the screening of drug-like libraries using an overnight high-throughput C. elegans assay, based on an automated infrared motility reader. As a proof of concept, we screened the “Pathogen Box” library, and identical results to a previous screen using Haemonchus contortus were obtained. We then screened an in-house library containing a diversity of compound families. Most active compounds had a conjugation of an unsaturation with an electronegative atom (N, O, or S) and an aromatic ring. Importantly, we identified symmetric arylidene ketones and aryl hydrazine derivatives as novel nematicides. Furthermore, one of these compounds, (1E,2E)-1,2-bis(thiophen-3-ylmethylene)hydrazine, was active as a nematicide at 25 µm, but innocuous to the vertebrate model zebrafish at 50 µm. Our results identified novel nematicidal scaffolds and illustrate the value of C. elegans in accelerating nematicide discovery using a nonlabor-intensive automated assay that provides a simple overnight readout. Full article

We estimated the probability of exposure of people to questing ticks, infected with bacterial agents of the tick—borne zoonoses—in Aosta Valley, western Alps, Italy. We collected ticks by dragging, and from collectors’ clothes in three hiking trails, which were divided into an internal path, with short vegetation, and an external part with taller grass. Dragging yielded 285 Ixodes ricinus nymphs and 31 adults, and two Dermacentor marginatus adults. Eleven I. ricinus nymphs and 9 adults were collected from collectors’ clothes. Borrelia burgdorferi s.l. was identified by PCR in 12 out of 30 I. ricinus nymphs (prevalence = 40.0%, 95% confidence interval = 22.5, 57.5). The prevalence of infection by Rickettsia spp. was 13.3% (95% CI = 1.2, 25.5). The probability of encountering at least one questing I. ricinus infected by each bacterial agent (probability of exposure, E) in 100 m² was obtained by combining the number of collected nymphs, the prevalence of infection by each bacterial agent, the frequency of passage by visitors, and the probability of tick attachment to people. The mean number of nymphs collected by dragging was greatest in the internal part of hiking trails (mean = 7.9). Conversely, E was greater in the external part (up to 0.14 for B. burgdorferi s.l., and 0.07 for Rickettsia spp.), due to a greater probability of tick attachment to people in relatively tall vegetation. Full article

In acute pancreatitis (AP), kidney injury (KI) can occur. Urinalysis and some urinary biomarkers have been proposed as prognostic tools in human AP. The aim of the study was to evaluate urinalysis and urinary GGT-to-urinary creatinine (uGGT/uCr) in canine AP and their association with possible outcomes. AP diagnosis was based on clinical and laboratory parameters, abnormal SNAP^® cPL™ test and compatible imaging. Urinary KI (uKI) was defined if dogs had urinary casts and/or proteinuria. Dogs (n = 70) were divided in survivors and non-survivors according to the 15-day outcome. Data were analyzed using statistical software. Seventy dogs were retrospectively included, of which 24 dogs (34%) died. uKI was detected in 36 dogs (37%) which was associated with mortality (p = 0.01, Odds ratio (OR) 3.9, 95% CI 1.3–11.56). Non-survivors showed higher dipstick bilirubin levels than survivors (p = 0.0022). By excluding active sediments, urine protein-to-creatinine ratio (UPC) ≥2 was associated with mortality (p = 0.001, OR 47.5, 95% CI 4–571.9). The uGGT/uCr was available in 40 dogs, although no association of this factor with any outcome was found. The UPC ≥2 can be a negative prognostic factor in canine AP and further studies on uGGT/uCr are warranted. Full article

Porcine respiratory disease complex (PRDC) is a polymicrobial syndrome that results from a combination of infectious agents, such as environmental stressors, population size, management strategies, age, and genetics. PRDC results in reduced performance as well as increased mortality rates and production costs in the pig industry worldwide. This review focuses on the interactions of two enveloped RNA viruses—porcine reproductive and respiratory syndrome virus (PRRSV) and swine influenza virus (SwIV)—as major etiological agents that contribute to PRDC within the porcine cellular innate immunity during infection. The innate immune system of the porcine lung includes alveolar and parenchymal/interstitial macrophages, neutrophils (PMN), conventional dendritic cells (DC) and plasmacytoid DC, natural killer cells, and γδ T cells, thus the in vitro and in vivo interactions between those cells and PRRSV and SwIV are reviewed. Likewise, the few studies regarding PRRSV-SwIV co-infection are illustrated together with the different modulation mechanisms that are induced by the two viruses. Alterations in responses by natural killer (NK), PMN, or γδ T cells have not received much attention within the scientific community as their counterpart antigen-presenting cells and there are numerous gaps in the knowledge regarding the role of those cells in both infections. This review will help in paving the way for future directions in PRRSV and SwIV research and enhancing the understanding of the innate mechanisms that are involved during infection with these viruses. Full article

Flaxseed is rich in α-linolenic acid and is used in broiler chicken diets to enrich tissues with n-3 fatty acids (FA). However, non-starch polysaccharides (NSP) in flaxseed decreases nutrient digestibility and limits the availability of n-3 FA. Addition of carbohydrase enzymes to flaxseed-based diets can decrease the anti-nutritive effects of NSP. We hypothesized that flaxseed and enzyme supplementation affect lipid content and alter expression of genes related to lipid metabolism in broiler liver. Five day-old broiler chicks were fed a corn-soybean basal diet with 0% flaxseed, a basal diet with 10% of flaxseed, or 10% flaxseed + 0.05% enzyme diet up to day 42 of growth. Total lipids, including long-chain (≥20C) n-3 FA and monounsaturated FA, were increased in flax-fed broiler livers. Enzyme addition reduced arachidonic acid and total long chain n-6 FA. These changes were similarly reflected in phosphatidylcholine lipid species. Dietary flax and enzyme treatments up-regulated PPARα target genes CPT1A and ACOX1 while reducing expression of de novo FA synthesis-related genes. This study concludes that flaxseed and enzyme supplementation in broiler diets enhances LC n-3 FA species, while reducing n-6 FA species in hepatic phospholipids (PL). Flaxseed-based diets changes the expression of genes involved in FA lipid metabolism without affecting growth or production performance in broilers. Full article

This review serves to summarize parasites found in Domesticated animals which were found in the Neo-Tropics. Indigenous domesticated Neo-tropical animals include South American camelids, (Lama gunacoa, Lama glama, Lama pacos, Vicuna vicuna), guinea pigs (Cavia porcellus), chinchillas (Chinchilla lanigera), turkeys (Meleagris gallopavo) and ducks (Cairina moschata, Anas platyrhynchos, Dendrocyga autumnalis). These animals were chosen due to their origin of existence (Neo-tropics) and over time these animals became domesticated and were distributed throughout the world. Over eighty (80) references were collected for this review and the papers spanned over eighty (80) years from 1934 to 2018. The gastrointestinal parasites reported for each animal were tabulated and their effects in the animal noted. Parasites reported in domesticated Neo-tropical animals had little to no effect on wild and free ranging animals with a few cases of illness and decreased productivity. The majority of articles viewed these animals as reservoir host which can infect humans and other domesticated livestock. It must also be noted that research done in the past did not focus on the effect these parasites had on these animals but only observed their potential as reservoirs for parasitic diseases. Full article

Dilated cardiomyopathy (DCM) is a myocardial disease of dogs and humans characterized by progressive ventricular dilation and depressed contractility and it is a frequent cause of heart failure. Conventional pharmacological therapy cannot reverse the progression of the disease and, in humans, cardiac transplantation remains the only option during the final stages of heart failure. Cytoprotective gene therapy with vascular endothelial growth factor-B₁₆₇ (VEGF-B₁₆₇) has proved an effective alternative therapy, halting the progression of the disease in experimental studies on dogs. The aim of this work was to test the tolerability and feasibility of intracoronary administration, under fluoroscopic guidance, of VEGF-B₁₆₇ carried by adeno-associated viral vectors in canine DCM patients. Ten patients underwent the gene delivery procedure. The intraoperative phase was well tolerated by all dogs. Clinical and echocardiographic assessments at 7- and 30-days post-procedure showed stable conditions compared to the pre-procedure phase. The results of this work indicate that intracoronary VEGF-B₁₆₇ gene delivery is feasible and tolerated in dogs with DCM. Further monitoring/investigations are ongoing to evaluate the effects of this therapy on disease progression. Full article

This study aimed to investigate collagen (Coll-I, III, IV) and elastin in canine normal prostate and prostate cancer (PC) using Picrosirius red (PSR) and Immunohistochemical (IHC) analysis. Eight normal prostates and 10 PC from formalin-fixed, paraffin-embedded samples were used. Collagen fibers area was analyzed with ImageJ software. The distribution of Coll-I and Coll-III was approximately 80% around prostatic ducts and acini, 15% among smooth muscle, and 5% surrounding blood vessels, in both normal prostate and PC. There was a higher median area of Coll-III in PC when compared to normal prostatic tissue (p = 0.001 for PSR and p = 0.05 for IHC). Immunostaining for Coll-IV was observed in the basal membrane of prostate acini, smooth muscle, blood vessels, and nerve fibers of normal and PC samples. Although there was no difference in Coll-IV area between normal tissue and PC, tumors with Gleason score 10 showed absence of Coll-IV, when compared to scores 6 and 8 (p = 0.0095). Elastic fibers were found in the septa dividing the lobules and around the prostatic acini of normal samples and were statistically higher in PC compared to normal tissue (p = 0.00229). Investigation of ECM components brings new information and should be correlated with prognosis in future studies. Full article

Laryngeal chondritis, or “Texel throat”, is a disease affecting the upper respiratory tract of sheep with breeds like the Texel appearing to be predisposed. Previous work suggests the conformation of these breeds of sheep may be predisposing these animals to laryngeal disease. This study evaluated the anatomy of the Texel sheep larynx and describes incidental pathology. Forty-three larynges from rams of the Texel and Bluefaced Leicester breeds of sheep were measured and photographed. A larynx from each breed was submitted for computed tomography (CT) and magnetic resonance imaging (MRI). Measurements, photography, CT, and MRI demonstrated a difference in the anatomy of the larynx between breeds and a higher proportion of Texel sheep had laryngeal lesions. This study supports the hypothesis that the anatomy of the Texel sheep could be pre-disposing the breed to laryngeal chondritis. Full article

Non-typhoid salmonellosis is a common and problematic foodborne zoonotic disease in which pork and pork products can be an important potential source of infection. To prevent this disease, important efforts to monitor the situation in the main source, livestock, are conducted in most developed countries. In the European Union, European Food Safety Agency (EFSA) and European Center for Disease Control (ECDC) compile information at the member-state level, even though important differences in production systems and surveillance systems exist. Here, Salmonella surveillance systems in one of the main sources of foodborne salmonellosis, swine, and humans in Spain were reviewed to identify potential gaps and discuss potential ways of integration under a “One-Health” approach. Despite the extensive information generated through the surveillance activities, source attribution can be only routinely performed through ad-hoc outbreak investigations, and national reports on human outbreaks do not provide sufficiently detailed information to gain a better understanding of the epidemiology of the pathogen. Human and animal monitoring of Salmonella would benefit from a better exchange of information and collaboration. Analysis of spatio-temporal trends in livestock and humans could help to identify likely sources of infection and to target surveillance efforts in areas with higher prevalence or where specific strains are found. Full article

As members of the public and the veterinary profession are increasingly concerned about animal welfare, there has been an increased scholarly interest in the attitudes of veterinarians and students toward animals, as these may impact human behavior, which ultimately impacts animal welfare. Here we investigated Italian veterinary students’ demographic data and perceptions about nonhuman animal welfare issues that might be predictive of their attitudes. A survey eliciting information about demographics, knowledge, experience, and perceptions regarding different categories of animals, and including the Animal Attitude Scale (AAS), was administered to undergraduate veterinary medicine students in three Italian universities. Data were analyzed using nonparametric tests, and a value of p < 0.05 was considered statistically significant. In total, 876 students completed the questionnaire, with females (75.1%) making up a majority of students in all years of the course. Although veterinary students showed pro-animal welfare attitudes (mean score = 64.20 ± 0.24 out of 100), the findings suggested that year of study, gender, and geographical location had a significant impact (p < 0.05). In this study, we found a set of factors that, either individually or combined, help predict a student’s attitude toward animal welfare issues, which will be useful in improving the curriculum strategy in veterinary education in Italy. Full article

Tibial Tuberosity Advancement (TTA) is a surgical technique based on a linear osteotomy that determines a cranial advancement of the tibial tuberosity in patients suffering from cranial cruciate ligament rupture (CCL). The aim is to neutralize the cranial tibial thrust (CTT) and to reach a 90° angle between the patellar tendon and the tibial plateau with a physiological knee extension of 135°. In our study, a Ti6AI4V ELI (Titanium Aluminium Vanadium) titanium scaffold for the Porous TTA, with excellent properties of osteointegration and osteoconduction when subjected to cyclic loading has been adopted. Based on the previous scientific work on an ovine model, the use of this type of porous scaffolds has subverted the previous models. Scaffold production technology is based on direct mechanical manufacturing called Electron Beam Melting (EBM). For this study, 41 dogs, different breeds, medium-large size, weighing between 10 and 80 kg, aged between 1 and 13 years, were enrolled. The inclusion criteria were based on clinical evaluations (different gaits), drawer test and tibial compression, LOAD score (Liverpool Osteoarthritis in Dogs questionnaire), radiographic diagnosis in sedation with a 135° positioning of the joint and baropodometric investigations (Stance Analyzer). The results show that Porous TTA is an excellent method for functional recovery of the knee joint following the partial and total rupture of the CCL. Full article

Pneumoperitoneum may induce important hemodynamic alterations in healthy subjects. Pulse pressure variation (PPV) is a hemodynamic parameter able to discriminate preload dependent subjects. Anesthesia records of dogs undergoing laparoscopy were retrospectively evaluated. The anesthetic protocol included acepromazine, methadone, propofol and isoflurane administered with oxygen under mechanical ventilation. The hemodynamic parameters were considered five minutes before (BASE) and ten minutes after (P10) the pneumoperitoneum. Based on the cardiac index (CI) variation, at P10, dogs were classified as sensitive (S group, CI ≤ 15%) and non-sensitive (NO-S group). Data were analyzed with the ANOVA test and the ROC curve (p < 0.05). Fifty-five percent of dogs (S) had a reduction of CI ≥ 15% at P10 (2.97 ± 1.4 L/min/m²) compared to BASE (4.32 ± 1.62 L/min/m²) and at P10 in the NO-S group (4.51 ± 1.41 L/min/m²). PPV at BASE was significantly higher in the S group (22.4% ± 6.1%) compared to the NO-S group (10.9% ± 3.3%). The ROC curve showed a threshold of PPV > 16% to distinguish the S and NO-S groups. PPV may be a valid predictor of the hemodynamic response to pneumoperitoneum in dogs. A PPV > 16% can identify patients that may require fluid administration before the creation of pneumoperitoneum. Full article

Mast cells (MCs) are involved in angiogenesis, tissue remodeling and immunomodulation in several human and animal tumors, although their exact role is still controversial. Since no information is available in canine prostate carcinoma (PC) and normal prostate tissues, the aims of this study were to evaluate the possible correlations between MC distribution, molecular expression and microvessel density (MVD) in normal prostatic tissue and proliferative disorders of the canine prostate. All samples (6 normal, 15 benign prostate hyperplasia-BPH, 8 PC) were stained with Toluidine Blue and immunohistochemically evaluated for tryptase, c-Kit (CD117) and CD31. Mast cell density (MCD) and MVD were quantified by the hot-spot method. MCD was significantly increased in periglandular/peritumoral areas, when compared with intraglandular/intratumoral areas, in all groups (p = 0.03). C-Kit expression was strongly associated with PC (ρ = 0.75 p = 0.03), whereas positive correlation between tryptase and c-Kit expression (ρ = 0.64 p = 0.01) was observed in periglandular areas of BPH. MVD showed a correlation with MCD in BPH (ρ = 0.54 p = 0.04). Our data support the importance of c-Kit in regulating MC proliferation. The predominant location of MCs in peritumoral areas of canine PC was similar to the human counterpart, in which PC cells are supposed to produce substances attracting MCs to the tumor microenvironment. Full article

In dogs, melanomas are relatively common tumors and the most common form of oral malignancy. Biological behavior is highly variable, usually aggressive, and frequently metastatic, with reported survival times of three months for oral or mucosal melanomas in advanced disease stages. Classical clinical management remains challenging; thus, novel and more efficacious treatment strategies are needed. Evidence-based medicine supports the role of the immune system to treat neoplastic diseases. Besides, immunotherapy offers the possibility of a precise medicinal approach to treat cancer. In recent years, multiple immunotherapeutic strategies have been developed, and are now recognized as a pillar of treatment. In addition, dogs represent a good model for translational medicine purposes. This review will cover the most relevant immunotherapeutic strategies for the treatment of canine malignant melanoma, divided among five different categories, namely, monoclonal antibodies, nonspecific immunotherapy activated by bacteria, vaccines, gene therapy, and lymphokine-activated killer cell therapy. Full article

Despite the characteristic species specificity of Papillomaviruses (PVs), the bovine papillomavirus (BPV) types 1, 2, and—more rarely—13, can cross-infect equids, where they are involved in the pathogenesis of sarcoid neoplasms. Sarcoids are locally invasive fibroblastic skin tumors that represent the most common skin neoplasms in horses worldwide. The transmission mechanism of BPV is still controversial in horses. Thus far, direct and indirect routes have been implicated, while vertical transmission has been suggested after the detection of viral DNA in the semen of healthy stallions. Testing of the blood and placenta of non-sarcoid baring mares and their respective foals revealed that the equine placenta can harbor BPV DNA, leading us to speculate a possible prenatal vertical DNA transmission in equids. Full article

The objective of the present study was the identification of farming practices in the production of turkeys for human consumption, and their ranking in terms of the occupational probability of exposure to antimicrobial resistant (AMR) bacteria, for farm workers. We gathered evidence and data from scientific literature, on risk factors for AMR in farmers, and on the prevalence of those hazards across farming phases. We administered semi-structured interviews to public and private veterinarians in Northern Italy, to obtain detailed information on turkey farming phases, and on working practices. Data were then integrated into a semi-quantitative Failure Modes and Effect Analysis (FMEA). Those working practices, which are characterized by direct contact with numerous animals, and which are carried out frequently, with rare use of personal protection devices resulted as associated with the greatest probability of exposure to AMR. For methicillin resistant Staphylococcus aureus (MRSA), these included vaccination and administration of any individual therapy, and removal and milling of litter, given the exposure of farmers to high dust level. Indeed, levels of occupational exposure to MRSA are enhanced by its transmission routes, which include direct contact with animal, as well as airborne transmission. Level of exposure to extended spectrum beta lactamase (ESBL) is more strictly associated with direct contact and the oral-fecal route. Consequently, exposure to ESBL resulted and associated with the routinely tipping over of poults turned on their back, and with the individual administration of therapies. Full article