Evaluation of Ki-67, goblet cell and MUC2 mucin RNA expression in dogs with lymphoplasmacytic and granulomatous colitis

Intestinal mucus barrier disruption may occur with chronic inflammatory enteropathies. The lack of studies evaluating mucus health in dogs with chronic colitis arises from inherent challenges with assessment of the intestinal mucus layer. It is therefore unknown if reduced goblet cell (GBC) numbers and/or mucin 2 (MUC2) expression, which are responsible for mucus production and secretion, correlate with inflammation severity in dogs with granulomatous colitis (GC) or lymphocytic-plasmacytic colitis (LPC). It is undetermined if Ki-67 immunoreactivity, which has been evaluated in dogs with small intestinal inflammation, similarly correlates to histologic severity in GC and LPC. Study objectives included comparing Ki-67 immunoreactivity, GBC population and MUC2 expression in dogs with GC, LPC and non-inflamed colon; and exploring the use of ribonucleic acid (RNAscope®) in-situ hybridization (ISH) to evaluate MUC2 expression in canine colon. Formalin-fixed endoscopic colonic biopsies were obtained from 48 dogs over an eight-year period. A blinded pathologist reviewed all biopsies. Dogs were classified into the GC (n=19), LPC (n=19) or no colitis (NC) (n=10) group based on final histopathological diagnosis. Ki-67 immunohistochemistry, Alcian-Blue/PAS staining to highlight GBCs, and RNAscope® ISH using customized canine MUC2-targeted probes were performed. At least five microscopic fields per dog were selected to measure Ki-67 labelling index (KI67%), GBC staining percentage (GBC%) and MUC2 expression (MUC2%) using image analysis software. Spearman's correlation coefficients were used to determine associations between World Small Animal Veterinary Association histologic score (WHS) and measured variables. Linear regression models were used to compare relationships between WHS with KI67%, GBC%, and MUC2%; and between GBC% and MUC2%. Median WHS was highest in dogs with GC. Median KI67% normalised to WHS was highest in the NC group (6.69%; range, 1.70–23.60%). Median GBC% did not correlate with colonic inflammation overall. Median MUC2% normalised to WHS in the NC group (10.02%; range, 3.05–39.09%) was two- and three-fold higher than in the GC and LPC groups respectively. With increased colonic inflammation, despite minimal changes in GBC% overall, MUC2 expression markedly declined in the LPC group (-27.4%; 95%-CI, −49.8, 5.9%) and mildly declined in the GC and NC groups. Granulomatous colitis and LPC likely involve different pathways regulating MUC2 expression. Decreased MUC2 gene expression is observed in dogs with chronic colitis compared to dogs without colonic signs. Changes in MUC2 expression appear influenced by GBC activity rather than quantity in GC and LPC.</p

The utility of diagnostic tests for immune-mediated hemolytic anemia

Background: A definitive diagnosis of immune‐mediated hemolytic anemia (IMHA) can be difficult to make. However, it is critical to differentiate IMHA from other causes of anemia due to the impact on prognosis and outcome for IMHA patients. Recently published American College of Veterinary Internal Medicine recommendations for the diagnosis of IMHA should be followed to concurrently confirm ongoing anemia, verify in vivo hemolysis, and detect anti‐erythrocyte antibodies. The reliability of immunologic IMHA tests varies depending on which test is used and how it is performed. Objectives: Our aims were to determine which tests are currently used in veterinary medicine to diagnose IMHA and review the utility of assays that have historically been used to diagnose IMHA. Methods: A short survey was designed to see which diagnostic tests for IMHA were currently being used by veterinary practices. The survey was distributed via listserves to veterinarians and veterinary technologists. A literature review was performed to report the utility of diagnostic tests for the diagnosis of IMHA. Results: Survey respondents indicated a variability in test protocols used to diagnose IMHA. Most respondents perform saline agglutination or Coombs’ tests to detect anti‐erythrocyte antibodies. Additional tests that can be used to support a diagnosis of IMHA are discussed in this review. Conclusions: A standardized diagnostic approach should be followed to differentiate IMHA from other causes of anemia. Test methodology can vary from one laboratory to another, and clinicians should be familiar with the procedures used by their laboratory

Prédiction des blessures des ischiojambiers en football à l'aide d'apprentissage automatique : étude préliminaire sur 284 footballeurs

peer reviewedThe hamstring injury is the number one injury diagnosis in football. One of the strategies for hamstring injury prevention is the identification of high-risk athletes. In this article, we implemented machine learning algorithms for injury risk estimation in 284 male footballers playing in 16 professional or semi-professional football teams from 3 countries. The predictors (input data) of the algorithms consisted of an athlete's baseline dataset, including hamstring injury history in the previous season and data measured during a maximum sprint of 30 m. The output data, binary, was the occurrence of hamstring injury. The three models used, logistic regression, random forests and AdaBoost, were compared to a dummy classifier. The results showed that it is possible, to a certain extent, to predict the occurrence of injury with these models. The comparison with the dummy classifier, when considering a set of metrics including F1-score, showed the interest of the three models used. Additionally, the relative importance of predictors can be measured, which can aid in understanding the predominant factors influencing injury. These results suggest avenues for hamstring injury prevention strategies

A retrospective multi‐center study of treatment, outcome, and prognostic factors in 34 dogs with disseminated aspergillosis in Australia

Background Disseminated aspergillosis (DA) in dogs has a guarded prognosis and there is a lack of a gold standard treatment protocol. Objective To retrospectively assess survival times and factors influencing survival times. Animals Dogs diagnosed with DA from January 2007 to June 2017. Methods Disseminated aspergillosis case data were retrieved from 13 Australian veterinary referral centers, with a diagnosis confirmed with culture or PCR. Factors influencing survival time after diagnosis were quantified using a Cox proportional hazards regression model. Results Thirty-four dogs met the study inclusion criteria. Twenty-two dogs were treated with antifungal treatment and 12 dogs received no antifungal treatment. Accounting for censoring of dogs that were either still alive on the date of data collection or were loss to follow-up, dogs treated with itraconazole alone (n = 8) had a median survival time (MST) of 63 (95% CI: 20−272) days compared to 830 (95% CI: 267-1259) days for the n = 14 dogs that received multimodal antifungal therapy

Hypovitaminosis D is associated with negative outcome in dogs with protein losing enteropathy: a retrospective study of 43 cases

Abstract Background Hypovitaminosis D has previously been shown to be prevalent amongst dogs with protein losing enteropathy (PLE). The hypothesis of this study was that Low 25-hydroxyvitamin D (25(OH) D) serum concentrations could be a risk factor for negative outcome in dogs with PLE. Forty-three dogs diagnosed with PLE (2005–2014) and which serum Vitamin D serum concentrations were collected and archived at −80 Degrees C were analyzed. Post-diagnostic communication with referring veterinarians was made to determine outcome of PLE dogss: Dogs which died due to PLE within 4 months after diagnosis (negative outcome group, n = 22) and dogs alive or which died due to another disease at the end point of the study (1 year after diagnosis, good outcome group, n = 21). Serum samples taken at the time of diagnosis were analysed for ionized calcium (iCa) concentrations and serum 25(OH) D concentration. Results Clinical (CCECAI) scores, age at PLE diagnosis, and iCa concentrations were not significantly different between dog groups. A significantly greater (p < 0.001) number of PLE dogs treated with hydrolyzed or elimination diet alone showed good outcome as compared to the PLE negative outcome group. Median serum 25(OH) D concentration was significantly (p = 0.017) lower in dogs with negative outcome versus PLE dogs with good outcome. Using logistic regression analysis, 25(OH) D serum concentration was shown to be a statistically significant factor for outcome determination. Cox regression analysis yielded a hazard ratio of 0.974 (95% CI 0.949, 0.999) per each one nmol/l increase in serum 25(OH) D concentration. Conclusions Low serum 25(OH) D concentration in PLE dogs was significantly associated with poor outcome. Further studies are required to investigate the clinical efficacy of Vitamin D (cholecalciferol) as a potential therapeutic agent for dogs with PLE

Clinical Guidelines for Fecal Microbiota Transplantation in Companion Animals

The Companion Animal Fecal Microbiota Transplantation (FMT) Consortium is an international group of veterinary experts that are currently performing FMT in dogs and cats. Based on available evidence and expert opinions, the Companion Animal FMT Consortium developed the first clinical guidelines for FMT in companion animals aimed at increasing the accessibility of this microbial-directed therapeutic in veterinary medicine. These clinical guidelines include recommendations and protocols for fecal donor screening, FMT product processing and preparation, and current FMT clinical indications and administration. These clinical guidelines are intended to be utilized by veterinarians in all practice types

Intestinal Small Cell Lymphoma: Are Dogs Big Cats?

Previously T-cell lymphomas of the gastrointestinal tract in human were classified as enteropathy-associated T-cell lymphoma (EATL) type I and type II. Type I is associated with celiac disease and characterized by large lymphocytes, whereas type II is not associated to enteropathies and characterized by small lymphocytes. In view of these differences, the nomenclature has been changed and EATL currently refers only to type I and type II has been renamed monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL).1 EATL has an aggressive clinical course and tumor cells most commonly have an αβ T-cell receptor phenotype. In comparison MEITL tumour cells express CD8, CD56, and megakaryocyte-associated tyrosine kinase. T-cell lymphoproliferative disorder (TLPD) is another type of small cell lymphoma described in the intestinal tract. This lymphoma has typically an indolent clinical course and commonly express CD8 and is negative for CD4 and CD56, Markers of T-cell lymphomas of the gastrointestinal tract have been much less extensively studied in cats and dogs and for this reason for the purpose of this lecture small cell lymphoma (SCL) will be used for neoplastic cells with nuclei smaller than 2 red blood cells in diameter and large cell lymphoma (LCL) for larger neoplastic cells. SCL characterized by infiltration of the intestinal mucosa by mature T-cells with variable epitheliotropism has been described for more than 10 years in cats. SCL has better outcome than other types of lymphoma in this species, with median survival times over 1.5 years.3 Although criteria have been described in cats to diagnose SCL, these are not as well defined in dogs. However, several recent studies support that SCL is also present in dogs and the clinical findings and outcome will be described in this presentation