Detection of Selected Equine Respiratory Pathogens in Stall Samples Collected at a Multi-Week Equestrian Show during the Winter Months.

The aim of this study was to use environmental sampling to determine the frequency of detection of selected equine respiratory viruses and bacteria in horses attending a multi-week equestrian show during the winter months. At four time points during showing, environmental sponge samples were collected from all stalls on the property and tested for the presence of equine herpesvirus-1 (EHV-1), EHV-2, EHV-4, equine influenza virus (EIV), equine rhinitis B virus (ERBV), Streptococcus equi ss. equi (S. equi), and S. equi ss. zooepidemicus (S. zooepidemicus) using real-time PCR (PCR). Environmental sponges were collected from all 53 barns by using one sponge for up to 10 stalls. Further, 2/53 barns were randomly selected for individual stall sampling in order to compare the results between individual and pooled stall samples. A total of 333/948 (35.13%, 95% CI 32.09-38.26%) pooled environmental stall sponges tested PCR-positive for at least one of the selected respiratory pathogens. Streptococcus zooepidemicus was the most commonly detected pathogen in pooled samples (28.69%, 95% CI 25.83-31.69%), followed by EHV-2 (14.45%, 95% CI 12.27-16.85%), EHV-4 (1.37%, 95% CI 0.73-2.33%), and a very small percentage of pooled stall sponges tested PCR-positive for EHV-1, ERBV, EIV, and S. equi. In individual samples, 171/464 (36.85%, 95% CI 32.45-41.42%) environmental stall sponges tested PCR-positive for at least one of the selected pathogens, following a similar frequency of pathogen detection as pooled samples. The detection frequency of true respiratory pathogens from environmental samples was higher during the winter months compared to previous studies performed during spring and summer, and this testing highlights that such pathogens circulate with greater frequency during the colder months of the year. The strategy of monitoring environmental stall samples for respiratory pathogens circumvents the often labor-intensive collection of respiratory secretions from healthy horses and allows for a more efficient assessment of pathogen buildup over time. However, environmental stall testing for respiratory pathogens should not replace proper biosecurity protocols, but it should instead be considered as an additional tool to monitor the silent circulation of respiratory pathogens in at-risk horses.

Multi-Centered Field Evaluation of a Salmonella spp. Point-of-Care PCR Assay Using Equine Feces and Environmental Samples.

The introduction of microfluidic card technology has opened the field for rapid point-of-care (POC) molecular assays, including fecal and environmental Salmonella spp. testing. The purpose of this study was to evaluate a novel POC PCR assay for the detection of Salmonella spp. in feces and environmental samples. A total of 143 fecal samples and 132 environmental samples were collected for POC PCR Salmonella spp. testing as well as qPCR testing. Each sample was inoculated into selenite broth and incubated for 18 to 24 hours. For the POC PCR assay, 14 µl of selenite broth were mixed with 126 µl of PCR reaction mix and pipetted into a microfluidic test card targeting the invA and ttrC gene of Salmonella enterica. For qPCR analysis, 200 µl of the selenite broth were processed for DNA purification and Salmonella spp. testing targeting the invA gene. The overall agreement between the POC PCR Salmonella spp. assay and qPCR assay was 88.1% for feces and 97.0% for environmental samples. Strong agreement and short turn-around-time make the POC device the first molecular diagnostic platform allowing detection of Salmonella spp. in a hospital setting without having to ship out samples to a veterinary diagnostic laboratory. The availability of an accurate POC PCR assay for the detection of Salmonella spp. will enhance the diagnostic capability of equine veterinarians to timely support a diagnosis of salmonellosis and also monitor the environment in order to reduce the risk of nosocomial infections.

Molecular Monitoring of EHV-1 in Silently Infected Performance Horses through Nasal and Environmental Sample Testing.

While the main goal in the management of an EHM outbreak focuses on identifying early clinical disease in order to physically separate infected horses, little effort is placed towards monitoring healthy horses. The assumption that EHV-1 shedding parallels clinical disease is erroneous, as subclinical shedders have been shown to be actively involved in viral spread. In an attempt to document the frequency of EHV-1 shedders and their impact on environmental contamination, we collected nasal swabs from 231 healthy horses and 203 environmental samples for the testing of EHV-1 by qPCR. Six horses and 28 stalls tested qPCR-positive for EHV-1. There was no association in the EHV-1 qPCR-positive status between nasal and stall swabs. While testing nasal secretions of healthy at-risk horses can detect active shedding at a specific time point, the testing of stall swabs allows to assess the temporal EHV-1 shedding status of a horse. The study results highlight the risk of subclinical EHV-1 shedders and stalls occupied by these horses as sources of infection for susceptible horses. The testing of individual stalls for the presence of EHV-1 may be a more practical approach than the collection of individual nasal swabs for the monitoring and early detection of the circulating virus. The results also highlight the need to improve the cleanliness and disinfection of stalls utilized by performance horses during show events.

Frequency of Detection of Respiratory Pathogens in Nasal Secretions From Healthy Sport Horses Attending a Spring Show in California.

The objective of this study was to determine detection frequency of respiratory viruses (equine influenza virus [EIV], equine herpesvirus-1 [EHV-1], EHV-2, EHV-4, EHV-5, equine rhinitis A virus [ERAV], ERBV) and bacteria (Streptococcus equi ss. equi[S. equi], S. equi ss. zooepidemicus[S. zooepidemicus]) in 162 nasal secretions and 149 stall swabs from healthy sport horses attending a spring show in California. Nasal and stall swabs were collected at a single time point and analyzed using qPCR. The detection frequency of respiratory pathogens in nasal secretions was 38.9% for EHV-2, 36.4% for EHV-5, 19.7% for S. zooepidemicus, 1.2% for ERBV, 0.6% for S. equi and 0% for EIV, EHV-1, EHV-4 and ERAV. The detection frequency of respiratory pathogens in stall swabs was 65.8% for S. zooepidemicus, 33.5% for EHV-2, 27.5% for EHV-5, 3.3% for EHV-1, 1.3% for EHV-4 and 0% for EIV, ERAV, ERBV and S. equi. Commensal viruses and bacteria were frequently detected in nasal secretions and stall swabs from healthy sport horses. This was in sharp contrast to the subclinical shedding of well-characterized respiratory pathogens. Of interest was the clustering of five EHV-1 qPCR-positive stalls from apparently healthy horses with no evidence of clinical spread. The results highlight the role of subclinical shedders in introducing respiratory pathogens to shows and their role in environmental contamination. The results also highlight the need to improve cleanliness and disinfection of stalls utilized by performance horses during show events.

Comparison of measured renal tumor size versus R.E.N.A.L. nephrometry score in predicting patient outcomes after robot-assisted laparoscopic partial nephrectomy.

Partial nephrectomy (PN) is a technically challenging procedure, making selection of appropriate patients paramount to a successful operation. To identify patients at increased risk of an adverse outcome after PN, there are a number of scoring systems available. The nephrometry score was initially described in a series of laparoscopic and open partial and radical nephrectomies. We compare the association of the nephrometry score with perioperative outcomes in a population of robot-assisted partial nephrectomies. A total of 119 patients were retrospectively reviewed. Correlation and regressional analysis was performed. We identified the separate variables R, E, N, and L to have limited correlation and no predictive value to patient outcomes. Nephrometry score and grade were found to have stronger correlation and predictive value than the individual components of the R.E.N.A.L. nephrometry score. Size of tumor measured on a continuous scale was found to have the strongest correlation and predictive value to outcomes. Outcomes predicted included operative time, length of stay, warm ischemia time, and entry into the collecting system.