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The objective of our study was to determine the clade affiliation of 116 contemporary equine Influenza A virus (EIV) isolates using pyrosequencing. The EIV isolates originated from horses with clinical signs of equine influenza and laboratory confirmation of EIV by real-time quantitative PCR (qPCR) in nasal secretions. Clade affiliation was performed on the basis of a single nucleotide polymorphism at 2 positions of the hemagglutinin 1 gene. Pyrosequencing was able to clearly classify EIV Florida sublineage prototype A/equine/Ohio/1/2003 and prototype A/equine/Richmond/1/2007 as clade 1 and 2, respectively. Out of the 116 EIV qPCR-positive samples, 113 (97.4%) were classified as belonging to clade 1 Florida sublineage, whereas 3 (2.6%) were classified as clade 2. All clade 1 EIV strains were detected in domestic horses, whereas the 3 clade 2 EIV strains originated from horses recently imported to the United States. Although clade 1 EIV strains are endemic in the United States, international transportation of horses represents a real risk in introducing clade 2 EIV strains into North America.

The combination of lower concentrations of antitumor drugs (carboplatin - CARB, doxorubicin - DOX, and methotrexate - MET) with propolis was investigated against canine osteosarcoma (spOS-2) and mesenchymal stem cells (MSC) in vitro. The mechanism of action in the combinations was analyzed. spOS-2 cells were incubated up to 72 h with propolis (50 μg/ml) alone or in combination with CARB (10-400 μmol/l), DOX (0.5-2 μmol/l) or MET (50-200 μmol/l). Cell viability was assessed by MTT assay, apoptosis/necrosis by flow cytometry, and MSC was incubated with the optimum combination. Propolis alone exerted no cytotoxic action against spOS-2 cells, whereas CARB (400, 200 and 100 μmol/l) exhibited the highest cytotoxic effects comparing to DOX and MET. The combination of propolis with the lowest concentrations of CARB led to better results comparing to CARB alone, which was not observed using DOX and MET. Apoptosis was involved in the action of propolis + CARB in spOS-2 cells. MSC were not affected by CARB/propolis, indicating that the cytotoxic action of the combination was specific to tumor cells but not to normal ones. Propolis improved the action of CARB against spOS-2 cells using lower concentrations of this drug, without affecting MSC. These findings are relevant and indicate a possible application of propolis in OSA treatment.

Background: Equine rhinitis B virus (ERBV) has been given little attention by practitioners compared to other respiratory viruses, mainly because of the lack of diagnostic modalities and association with clinical disease. The objective of the study was to determine the frequency of detection of ERBV in nasal secretions from 6568 horses with acute onset of respiratory signs. Methods: ERBV-positive qPCR results from nasal secretions submitted to a molecular diagnostic laboratory from 2013 to 2019 were reviewed. Results: A total of 333 ERBV qPCR-positive samples (5.1%) were detected with increasing yearly frequency since the introduction of the assay in 2013. In comparison, only three of 356 (0.8%) healthy horses tested qPCR-positive for ERBV. Median age for ERBV qPCR-positive horses was 3 years of age, and fever, coughing and nasal discharge were the most common signs reported. Further, co-infections with other respiratory pathogens were reported in 73 (21.9%) of ERBV qPCR-positive samples. Conclusion:ERBV is a commonly detected respiratory virus from nasal secretions of young horses presenting with fever, nasal discharge and coughing.

The use of renal biopsy through laparoscopy is increasingly present both in human and veterinary medicine. However, both techniques require skill and training to make the operator capable to do it. The learning curve allows the quantitative and qualitative assessment of the number of attempts and minimum time for the surgical procedure. The objective included establish the learning curve for laparoscopy-guided kidney biopsy procedures in dog and pig corpses. Six dogs and six pigs corpses weighing less than 10 kg were used for this study. All corpses underwent kidney biopsy performed through laparoscopy. Twenty-four operators, two per animal, performed 20 renal biopsies each (10 for each kidney), with 480 collection-procedures in total. Duration and difficulty of the procedure and the biopsy sample quality were evaluated and statistical analysis was performed using a mixed regression model with a random effect of individuals and multivariate analysis of data. There were 91.5% of the samples that were adequate for evaluation. There was no significant difference in the number of glomeruli or cortex percentage considering the attempts in either species, demonstrating the operator’s ability since first collection. Swine samples showed higher amounts of renal cortex than canine samples. The procedure duration was shorter as more attempts were performed in dogs and pigs. From the fourth repetition, the professional reached a plateau for the variable related to ‘collection’, and from the second, the professional presented uniform duration for ‘sample storage’. Operators of the swine model acquired more agility than the dog ones. The variable ‘difficulty’ decreased as more repetitions were performed, reaching a plateau in the sixth attempt. Seven renal biopsies laparoscopy-guided are required for an operator to be considered ‘capable’ to perform the procedure in the referred species included. The learning curve for image-guided kidney biopsy procedures improves the implementation of this technique and benefits patients that undergo this procedure.

OBJECTIVE Mesenchymal stromal (stem) cells (MSCs) have been studied to treat many common orthopedic injuries in horses. However, there is limited information available on when and how to use this treatment effectively. The aim of this retrospective study is to report case features, treatment protocols, and clinical outcomes in horses treated with MSCs. ANIMALS 65 horses presenting with tendinous, ligamentous, and articular injuries, and treated with MSCs prepared by a single laboratory between 2016 and 2019. Outcome information was available for 26 horses. PROCEDURES Signalment, clinical signs, diagnostic methods, treatment protocol features (prior and concurrent therapies, cell origin, dose, application site and number), and effective outcomes were analyzed. The analysis was focused on comparing the effect of different MSC treatment protocols (eg, autologous vs allogeneic) on outcome rather than the effectiveness of MSC treatment. RESULTS MSC treatment resulted in 59.1% (clinical lameness) to 76.9% (imaging structure) improvement in horses with diverse ages, breeds, sex, and lesions. The use of other therapeutic methods before MSC application (eg, anti-inflammatories, shockwave, laser, icing, resting, bandage and stack wrap, intra-articular injections, and/or surgical debridement) was shown to be statistically more effective compared to MSCs used as the primary therapeutic procedure (P < .05). Autologous versus allogeneic treatment outcomes were not significantly different. CLINICAL RELEVANCE A prospective MSC treatment study with standardization and controls to evaluate the different features of MSC treatment protocols is needed. The various case presentations and treatment protocols evaluated can be used to inform practitioners who are currently using MSCs in clinical practice.

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