Transcriptomic analysis of equine chorioallantois reveals immune networks and molecular mechanisms involved in nocardioform placentitis.

Nocardioform placentitis (NP) continues to result in episodic outbreaks of abortion and preterm birth in mares and remains a poorly understood disease. The objective of this study was to characterize the transcriptome of the chorioallantois (CA) of mares with NP. The CA were collected from mares with confirmed NP based upon histopathology, microbiological culture and PCR for Amycolatopsis spp. Samples were collected from the margin of the NP lesion (NPL, n = 4) and grossly normal region (NPN, n = 4). Additionally, CA samples were collected from normal postpartum mares (Control; CRL, n = 4). Transcriptome analysis identified 2892 differentially expressed genes (DEGs) in NPL vs. CRL and 2450 DEGs in NPL vs. NPN. Functional genomics analysis elucidated that inflammatory signaling, toll-like receptor signaling, inflammasome activation, chemotaxis, and apoptosis pathways are involved in NP. The increased leukocytic infiltration in NPL was associated with the upregulation of matrix metalloproteinase (MMP1, MMP3, and MMP8) and apoptosis-related genes, such as caspases (CASP3 and CASP7), which could explain placental separation associated with NP. Also, NP was associated with downregulation of several placenta-regulatory genes (ABCG2, GCM1, EPAS1, and NR3C1), angiogenesis-related genes (VEGFA, FLT1, KDR, and ANGPT2), and glucose transporter coding genes (GLUT1, GLUT10, and GLUT12), as well as upregulation of hypoxia-related genes (HIF1A and EGLN3), which could elucidate placental insufficiency accompanying NP. In conclusion, our findings revealed for the first time, the key regulators and mechanisms underlying placental inflammation, separation, and insufficiency during NP, which might lead to the development of efficacious therapies or diagnostic aids by targeting the key molecular pathways.

Emergence of Resistance to Macrolides and Rifampin in Clinical Isolates of Rhodococcus equi from Foals in Central Kentucky, 1995 to 2017.

The objective of this study was to determine the prevalence of Rhodococcus equi strains resistant to macrolides and rifampin over time in clinical samples from foals submitted to diagnostic laboratories in central Kentucky. We performed a retrospective observational study of all clinical samples from foals that were submitted to veterinary diagnostic laboratories in Kentucky between January 1995 and December 2017. Samples were included if the R. equi bacterium was cultured and tested for in vitro susceptibility to erythromycin or rifampin. In vitro susceptibility testing to erythromycin was available for 2,169 isolates of R. equi, while susceptibility testing to both erythromycin and rifampin was available for 1,681 isolates. Rifampin resistance was first detected in 2000, and erythromycin resistance was first detected in 2004. Between 1995 and 2006, the proportion of resistant isolates of R. equi was 0.7% for erythromycin and 2.3% for rifampin. There was a significant (P < 0.001) increase in the proportion of resistant R. equi between 2007 and 2017, with 13.6% of isolates being resistant to erythromycin and 16.1% being resistant to rifampin. Between 2007 and 2017, isolates of R. equi resistant to erythromycin or rifampin were significantly less likely to be isolated from feces than from the respiratory tract, other soft tissues, or musculoskeletal infections. The considerable increase in the prevalence of isolates of R. equi resistant to macrolides and rifampin since 2007 is of concern for both human and animal health.

In vitro culture of Parascaris equorum larvae and initial investigation of parasite excretory-secretory products.

Currently, diagnosis of Parascaris equorum infection in equids is limited to patent infections. The goals of this study were to culture P. equorum larvae in vitro and identify excretory-secretory (ES) products for prepatent diagnostic testing. Parascaris equorum L2/L3 larvae were hatched and cultured for up to 3 weeks for ES product collection. Fifth stage (L5) P. equorum were also cultured for ES product collection. Examination of ES fractions by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and silver stain revealed L2/L3 products ranging from 12-94 kDa and L5 products ranging from 12-189 kDa. Western blot analyses were conducted using polyclonal antibodies produced against P. equorum or Baylisascaris procyonis L2/L3 ES products, sera from rabbits inoculated with B. procyonis or Toxocara canis eggs, and sera from animals naturally infected with P. equorum or T. canis. Western blot results indicated parasite antigens migrating at 19 and 34 kDa may be useful for specifically detecting P. equorum infections.

Effects of location for collection of air samples on a farm and time of day of sample collection on airborne concentrations of virulent Rhodococcus equi at two horse breeding farms.

OBJECTIVE: To determine whether airborne concentrations of virulent Rhodococcus equi at 2 horse breeding farms varied on the basis of location, time of day, and month. SAMPLE POPULATION: 2 farms in central Kentucky with recurrent R equi-induced pneumonia in foals. PROCEDURES: From February through July 2008, air samples were collected hourly for a 24-hour period each month from stalls and paddocks used to house mares and their foals. Concentrations of airborne virulent R equi were determined via a modified colony immunoblot technique. Differences were compared by use of zero-inflated negative binomial methods to determine effects of location, time, and month. RESULTS: Whether mares and foals were housed predominantly in stalls or paddocks significantly affected results for location of sample collection (stall vs paddock) by increasing airborne concentrations of virulent R equi at the site where horses were predominantly housed. Airborne concentrations of virulent R equi were significantly higher from 6:00 pm through 11:59 pm than for the period from midnight through 5:59 am. Airborne concentrations of virulent R equi did not differ significantly between farms or among months. CONCLUSIONS AND CLINICAL RELEVANCE: Airborne concentrations of virulent R equi were significantly increased when horses were predominantly housed at the site for collection of air samples (ie, higher in stalls when horses were predominantly housed in stalls and higher in paddocks when horses were predominantly housed in paddocks). Concentrations of virulent R equi among air samples collected between the hours of 6:00 am and midnight appeared similar.

A proposal to leverage high-quality veterinary diagnostic laboratory large data streams for animal health, public health, and One Health.

Test data generated by ~60 accredited member laboratories of the American Association of Veterinary Laboratory Diagnosticians (AAVLD) is of exceptional quality. These data are captured by 1 of 13 laboratory information management systems (LIMSs) developed specifically for veterinary diagnostic laboratories (VDLs). Beginning ~2000, the National Animal Health Laboratory Network (NAHLN) developed an electronic messaging system for LIMS to automatically send standardized data streams for 14 select agents to a national repository. This messaging enables the U.S. Department of Agriculture to track and respond to high-consequence animal disease outbreaks such as highly pathogenic avian influenza. Because of the lack of standardized data collection in the LIMSs used at VDLs, there is, to date, no means of summarizing VDL large data streams for multi-state and national animal health studies or for providing near-real-time tracking for hundreds of other important animal diseases in the United States that are detected routinely by VDLs. Further, VDLs are the only state and federal resources that can provide early detection and identification of endemic and emerging zoonotic diseases. Zoonotic diseases are estimated to be responsible for 2.5 billion cases of human illness and 2.7 million deaths worldwide every year. The economic and health impact of the SARS-CoV-2 pandemic is self-evident. We review here the history and progress of data management in VDLs and discuss ways of seizing unexplored opportunities to advance data leveraging to better serve animal health, public health, and One Health.

Antibiotic prescription practices and opinions regarding antimicrobial resistance among veterinarians in Kentucky, USA.

BACKGROUND: Inappropriate antimicrobial use (AMU) is a global concern. Opinions of veterinarians regarding AMU and its role in the development of antimicrobial resistance (AMR) may influence their prescription practices. It is important to understand these opinions, prescription practices and their potential impact on the development of AMR in order to guide efforts to curb the problem. Therefore, the objective of this study was to investigate the antimicrobial prescription practices and opinions of veterinarians in Kentucky regarding AMU and AMR. METHODS: This cross-sectional study used a 30-question survey questionnaire administered to veterinarians who were members of the Kentucky Veterinary Medical Association. Survey responses from 101 participants were included in the study. Descriptive statistics were computed and associations between categorical variables assessed using Chi-square or Fisher's exact tests. Firth logistic models were used to investigate predictors of "Compliance with prescription policies" and "Cost of antimicrobial affects prescription decisions". RESULTS: Almost all (93%) respondents indicated that improper AMU contributed to selection for AMR. A total of 52% of the respondents believed that antimicrobials were appropriately prescribed, while the remaining 48% believed that antimicrobials were inappropriately prescribed. Significant predictors of compliance with prescription policies were availability of prescription policy at the veterinary facility (Odds Ratio (OR) = 4.2; p<0.001) and over-prescription (OR = 0.35; p = 0.025). Similarly, significant predictors of cost of antimicrobials affecting prescription decisions were lack of post-graduate training (OR = 8.3; p = 0.008) and practice type, with large animal practices having significantly lower odds of the outcome (OR = 0.09; p = 0.004) than small animal practices. CONCLUSION: Most veterinarians indicated that improper AMU contributed to selection for AMR. Since the odds of compliance with prescription policies were 4-times higher among veterinarians working at facilities that had prescription policies compared to those at facilities that didn't, more veterinary facilities should be encouraged to adopt prescription policies to help improve compliance and reduce AMR. Veterinarians would also benefit from continued professional education to help improve prescription practices, antimicrobial stewardship and curb AMR.

Identification of a Ruminant Origin Group B Rotavirus Associated with Diarrhea Outbreaks in Foals.

Equine rotavirus group A (ERVA) is one of the most common causes of foal diarrhea. Starting in February 2021, there was an increase in the frequency of severe watery to hemorrhagic diarrhea cases in neonatal foals in Central Kentucky. Diagnostic investigation of fecal samples failed to detect evidence of diarrhea-causing pathogens including ERVA. Based on Illumina-based metagenomic sequencing, we identified a novel equine rotavirus group B (ERVB) in fecal specimens from the affected foals in the absence of any other known enteric pathogens. Interestingly, the protein sequence of all 11 segments had greater than 96% identity with group B rotaviruses previously found in ruminants. Furthermore, phylogenetic analysis demonstrated clustering of the ERVB with group B rotaviruses of caprine and bovine strains from the USA. Subsequent analysis of 33 foal diarrheic samples by RT-qPCR identified 23 rotavirus B-positive cases (69.69%). These observations suggest that the ERVB originated from ruminants and was associated with outbreaks of neonatal foal diarrhea in the 2021 foaling season in Kentucky. Emergence of the ruminant-like group B rotavirus in foals clearly warrants further investigation due to the significant impact of the disease in neonatal foals and its economic impact on the equine industry.

Application of an automated surveillance-data-analysis system in a laboratory-based early-warning system for detection of an abortion outbreak in mares.

OBJECTIVE: To develop an early-warning automated surveillance-data-analysis system for early outbreak detection and reporting and to assess its performance on an abortion outbreak in mares in Kentucky. SAMPLE POPULATION: 426 data sets of abortions in mares in Kentucky during December 2000 to July 2001. PROCEDURES: A custom software system was developed to automatically extract and analyze data from a Laboratory Information Management System database. The software system was tested on data on abortions in mares in Kentucky reported between December 1, 2000, and July 31, 2001. The prospective space-time permutations scan statistic, proposed by Kulldorff, was used to detect and identify abortion outbreak signals. RESULTS: Results indicated that use of the system would have detected the abortion outbreak approximately 1 week earlier than traditional surveillance systems. However, the geographic scale of analysis was critical for highest sensitivity in outbreak detection. Use of the lower geographic scale of analysis (ie, postal [zip code]) enhanced earlier detection of significant clusters, compared with use of the higher geographic scale (ie, county). CONCLUSIONS AND CLINICAL RELEVANCE: The automated surveillance-data-analysis system would be useful in early detection of endemic, emerging, and foreign animal disease outbreaks and might help in detection of a bioterrorist attack. Manual analyses of such a large number of data sets (ie, 426) with a computationally intensive algorithm would be impractical toward the goal of achieving near real-time surveillance. Use of this early-warning system would facilitate early interventions that should result in more positive health outcomes.

Association of soil concentrations of Rhodococcus equi and incidence of pneumonia attributable to Rhodococcus equi in foals on farms in central Kentucky.

OBJECTIVE: To determine whether soil concentrations of total or virulent Rhodococcus equi differed among breeding farms with and without foals with pneumonia caused by R equi. SAMPLE POPULATION: 37 farms in central Kentucky. Procedures-During January, March, and July 2006, the total concentration of R equi and concentration of virulent R equi were determined by use of quantitative bacteriologic culture and a colony immunoblot technique, respectively, in soil specimens obtained from farms. Differences in concentrations and proportion of virulent isolates within and among time points were compared among farms. RESULTS: Soil concentrations of total or virulent R equi did not vary among farms at any time point. Virulent R equi were identified in soil samples from all farms. Greater density of mares and foals was significantly associated with farms having foals with pneumonia attributable to R equi. Among farms with affected foals, there was a significant association of increased incidence of pneumonia attributable to R equi with an increase in the proportion of virulent bacteria between samples collected in March and July. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that virulent R equi were commonly recovered from soil of horse breeding farms in central Kentucky, regardless of the status of foals with pneumonia attributable to R equi on each farm. The incidence of foals with pneumonia attributable to R equi can be expected to be higher at farms with a greater density of mares and foals.

Temporal trends and predictors of antimicrobial resistance among Staphylococcus spp. isolated from canine specimens submitted to a diagnostic laboratory.

BACKGROUND: Resistance to commonly used antimicrobials is a growing concern in both human and veterinary medicine. Understanding the temporal changes in the burden of the problem and identifying its determinants is important for guiding control efforts. Therefore, the objective of this study was to investigate temporal patterns and predictors of antimicrobial resistance among Staphylococcus spp. isolated from canine specimens submitted to the University of Kentucky Veterinary Diagnostic Laboratory (UKVDL) between 1993 and 2009. METHODS: Retrospective data of 4,972 Staphylococcus isolates assessed for antimicrobial susceptibility using the disk diffusion method at the UKVDL between 1993 and 2009 were included in the study. Temporal trends were assessed for each antimicrobial using the Cochran-Armitage trend test. Logistic regression models were used to investigate predictors of antimicrobial resistance (AMR) and multidrug resistance (MDR). RESULTS: A total of 68.2% (3,388/4,972) Staphylococcus isolates were S. intermedius group (SIG), 18.2% (907/4,972) were coagulase-negative staphylococci (CoNS), 7.6% (375/4,972) were S. aureus, 5.8% (290/4,972) were S. hyicus, and S. schleiferi subsp. coagulans comprised 0.2% (12/4,972) of the isolates. The overall percentage of AMR and MDR were 77.2% and 25.6%, respectively. The highest levels of AMR were seen in CoNS (81.3%; 737/907), S. aureus (80.5%; 302/375), and SIG (77.6%; 2,629/3388). The lowest levels of AMR were observed in S. hyicus (57.9%; 168/290) and S. schleiferi subsp. coagulans (33.3%; 4/12). Overall, AMR and MDR showed significant (p<0.001) decreasing temporal trends. Significant temporal trends (both increasing and decreasing) were observed among 12 of the 16 antimicrobials covering 6 of the 9 drug classes assessed. Thus, significant increasing temporal trends in resistance were observed to ß-lactams (p<0.001) (oxacillin, amoxicillin-clavulanate, cephalothin, and penicillin (p = 0.024)), aminoglycosides (p<0.001) (gentamicin, and neomycin), bacitracin (p<0.001), and enrofloxacin (p<0.001). In contrast, sulfonamide (p<0.001) (sulfadiazin) and tetracycline (p = 0.010) resistant isolates showed significant decreasing temporal trends in AMR. Staphylococcus spp., geographic region, and specimen source were significant predictors of both AMR and MDR. CONCLUSIONS: Although not unexpected nor alarming, the high levels of AMR to a number of antimicrobial agents and the increasing temporal trends are concerning. Therefore, continued monitoring of AMR among Staphylococcus spp. is warranted. Future studies will need to identify local factors responsible for the observed geographic differences in risk of both AMR and MDR.

Equine antibody response to larval Parascaris equorum excretory-secretory products.

Parascaris equorum is an intestinal nematode of foals and young horses that can produce mild to severe pathology. Current diagnosis is limited to detection of patent infections, when parasite eggs are identified during fecal examinations. This study examined the use of larval P. equorum excretory-secretory (ES) products in a western blot test for diagnosis of prepatent equine P. equorum infection. Sera from adult mares negative for patent P. equorum infections, foals prior to consuming colostrum, and P. equorum infected foals were used as controls in this study. Study samples included sera from 18 broodmares prior to parturition and sera from their foals throughout the process of natural infection. Sera from study horses were examined for IgG(T) antibody recognition of ES products. Foals naturally infected with P. equorum possessed IgG(T) antibodies against 19kDa, 22kDa, 26kDa, and 34kDa ES products. However, passive transfer of colostral antibodies from mares was shown to preclude the use of the crude larval ES product-based western blot test for diagnosis of prepatent P. equorum infections in foals.

Discrimination of healthy versus sick steers by means of continuous remote monitoring of animal activity.

OBJECTIVE: To test a unique electronic ear tag designed to collect movement data to determine whether physical activity of sick steers differed from that of healthy steers. ANIMALS: 206 steers. PROCEDURES: Physical activity in 2 groups of steers during November and December of 2010 (101 steers; the tag of 1 steer failed, and thus that steer was removed from the study, which resulted in data for 100 steers) and 2011 (105 steers) was monitored with an electronic ear tag device with an on-board triple-axis accelerometer. The accelerometer recorded motion in all 3 axes in the form of counts per minute. A radio-frequency transmitter on the ear tag delivered serial packets of motion data to a local server. An algorithm was developed to analyze the activity data to determine whether this technique could be used to assess health status with high accuracy. RESULTS: Steers that became sick had significantly fewer activity counts (approx 25% fewer), compared with the activity counts of steers that remained healthy the entire time. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, automated detection of health status in growing cattle was feasible through remote monitoring of animal activity. Early identification of sick animals should lead to improved health outcomes, increased marketability, and improved animal well-being and help to minimize the use of antimicrobials that could contribute to resistant bacteria.

Frequency of Corynebacterium pseudotuberculosis infection in horses across the United States during a 10-year period.

OBJECTIVE: To quantify the number of horses with Corynebacterium pseudotuberculosis infection identified in the United States from January 2003 through December 2012. DESIGN: Cross-sectional study. SAMPLE: State veterinary diagnostic laboratory records of 2,237 C pseudotuberculosis culture-positive samples from horses. PROCEDURES: 44 state veterinary diagnostic laboratories throughout the United States were invited by mail to participate in the study. Data requested included the number of C pseudotuberculosis culture-positive samples from horses identified per year, geographic location from which the C pseudotuberculosis culture-positive sample was submitted, month and year of sample submission, breed and age of horses, and category of clinical manifestation (ie, internal infection, external infection, or ulcerative lymphangitis). RESULTS: Of the 44 invited laboratories, 15 agreed to participate and provided data on affected horses from 23 states. The proportion of C pseudotuberculosis culture-positive samples submitted during 2011 through 2012 (1,213/2,237 [54%]) was significantly greater than that for the period from 2003 through 2010 (1,024/2,237 [46%]). Corynebacterium pseudotuberculosis was recovered from horses in states where the disease has not been previously recognized as endemic. Affected horses were identified year-round. The greatest proportion of C pseudotuberculosis culture-positive samples was identified during November, December, and January (789/2,237 [35%]). No significant association between the clinical form of disease and age or breed of horse was observed. CONCLUSIONS AND CLINICAL RELEVANCE: The occurrence of C pseudotuberculosis infection in horses increased during the 10-year period, and affected horses were identified throughout the United States. Further studies to determine changes in annual incidence and to identify potential changing climatic conditions or vector populations associated with disease transmission are warranted to help control the occurrence and spread of C pseudotuberculosis infection in horses.

Beta-hemolytic Streptococcus spp. from horses: a retrospective study (2000-2010).

The goal of this retrospective study was to have a comprehensive picture of the ß-hemolytic streptococci of horses including tissue/organ distributions and susceptibility patterns against specific antimicrobials between January 1, 2000 and December 31, 2010. A total of 2,497 ß-hemolytic streptococci were isolated from 2,391 cases, of which Streptococcus equi subsp. zooepidemicus was the most frequent isolate (72.0%). Other species isolated were Streptococcus dysgalactia subsp. equisimilis (21.3%), Streptococcus equi subsp. equi (5.8%), and unidentified ß-hemolytic streptococci (0.9%). As expected, S. equi was mostly isolated from lymph node abscesses and the respiratory tract in foals and adult horses. Streptococcus equi subsp. zooepidemicus and S. equisimilis were mostly isolated from placenta, fetal tissues, and genital tract of horses; S. zooepidemicus and S. equisimilis were also recovered in significant numbers from a number of other organs including lung, liver, brain, kidney, and joints, indicating a much broader tissue tropism than S. equi. In addition, more than 1 Streptococcus spp. was recovered in 106 cases, indicating the co-existence of these bacteria in some horses. This data also suggested that S. equisimilis is a major bacterial agent of horses, contrary to present knowledge. Based on Kirby-Bauer antimicrobial susceptibility data, streptococci were found to be generally susceptible to cephalothin, erythromycin, nitrofurantoin, penicillin, and ticarcillin and clavulanate. Resistance to antimicrobials has not developed over the years, except for gentamicin and tetracycline against S. equisimilis.

An investigation of a recent outbreak of nocardioform placentitis caused abortions in horses.

Nocardioform placentitis associated with gram positive branching actinomycetes caused a record number of abortions in mares diagnosed by the University of Kentucky Veterinary Diagnostic Laboratory (UKVDL) affecting the 2011 foal crop (2011 foal crop: the cohort of foals conceived during the 2010 breeding season). The goal of the present study is to make a comprehensive analysis of this outbreak in terms of frequencies of the bacteria causing nocardioform placentitis mediated abortions and to investigate the ages of fetuses, abortion months and breeding times. In the present study, characteristic slow-growing, pungent/soil odor gram positive branching actinomycetes were recovered in high numbers in placental specimens in 76 abortion cases diagnosed as nocardioform placentitis by pathologists. To determine the type of actinomycetes responsible for the abortions, PCR assays were performed on the gram positive branching bacilli. The most prominent actinomycetes species were Amycolatopsis spp. (37 cases, 48.7%) and Crossiella equi (C. equi) (22 cases, 28.9%). Six cases (7.9%) contained both Amycolatopsis spp., and C. equi. 10 isolates were unidentified by PCR assays and shown to have high DNA sequence homology to Streptomyces species, Microbacterium species, Nocardia species and Allokutzneria species, as evidenced by 16 rRNA gene sequencing and phylogenetic analysis. Nocardioform placentitis related abortions occurred mostly between December 2010 and April 2011 happening exclusively in the last trimester. Breeding time of aborted pregnancies ranged from March 2010 to July 2010, suggesting that if transmission of the actinomycetes agents occurred during breeding, it was not related to a specific season.

Antibiotic susceptibility patterns of Crossiella equi and Amycolatopsis species causing nocardioform placentitis in horses.

Nocardioform actinomycetes are significant causes of placentitis and abortions in horses. In the current study, antimicrobial susceptibility patterns of 38 Amycolatopsis spp. and 22 Crossiella equi isolates, the most common nocardioform actinomycetes causing placentitis in horses, were evaluated. Antimicrobial susceptibilities of these isolates were tested by broth microdilution method in a commercial system, which was designed for Nocardia spp., fast-growing Mycobacterium spp., and other aerobic actinomycetes. The minimum inhibitory concentration required to inhibit the growth of 90% of organisms (MIC(90)) of the following antibiotics tested for Amycolatopsis spp. were: 4 µg/ml for linezolid, trimethophrim-sulfametaxazole (TMP-SMX), and ciprofloxacin; 8 µg/ml for ceftriaxone, doxycycline, and minocycline; 16 µg/ml for amoxicillin-clavulanic acid, clarithromycin, and imipenem; >16 µg/ml for tobramycin; 32 µg/ml for amikacin and cefepime; and 128 µg/ml for cefoxitin. The MIC(90) levels for C. equi were 0.25 µg/ml for doxycycline; ≤1 µg/ml for minocycline; 2 µg/ml for linezolid and TMP-SMX; 4 µg/ml for ciprofloxacin; 8 µg/ml for amoxicillin-clavulanic acid, ceftriaxone, and imipenem; 16 µg/ml for clarithromycin; >16 µg/ml for tobramycin; 32 µg/ml for cefepime; >64 µg/ml for amikacin; and 128 µg/ml for cefoxitin.

Association of airborne concentration of virulent Rhodococcus equi with location (stall versus paddock) and month (January through June) on 30 horse breeding farms in central Kentucky.

OBJECTIVE: To determine whether the concentration of airborne virulent Rhodococcus equi varied by location (stall vs paddock) and month on horse farms. SAMPLE: Air samples from stalls and paddocks used to house mares and foals on 30 horse breeding farms in central Kentucky. PROCEDURES: Air samples from 1 stall and 1 paddock were obtained monthly from each farm from January through June 2009. Concentrations of airborne virulent R equi were determined via a modified colony immunoblot assay. Random-effects logistic regression was used to determine the association of the presence of airborne virulent R equi with location from which air samples were obtained and month during which samples were collected. RESULTS: Of 180 air samples, virulent R equi was identified in 49 (27%) and 13 (7%) obtained from stalls and paddocks, respectively. The OR of detecting virulent R equi in air samples from stalls versus paddocks was 5.2 (95% confidence interval, 2.1 to 13.1). Of 60 air samples, virulent R equi was identified in 25 (42%), 18 (30%), and 6 (10%) obtained from stalls during January and February, March and April, and May and June, respectively. The OR of detecting virulent R equi from stall air samples collected during May and June versus January and February was 0.22 (95% confidence interval, 0.08 to 0.63). CONCLUSIONS AND CLINICAL RELEVANCE: Foals were more likely to be exposed to airborne virulent R equi when housed in stalls versus paddocks and earlier (January and February) versus later (May and June) during the foaling season.

OAS1 polymorphisms are associated with susceptibility to West Nile encephalitis in horses.

West Nile virus, first identified within the United States in 1999, has since spread across the continental states and infected birds, humans and domestic animals, resulting in numerous deaths. Previous studies in mice identified the Oas1b gene, a member of the OAS/RNASEL innate immune system, as a determining factor for resistance to West Nile virus (WNV) infection. A recent case-control association study described mutations of human OAS1 associated with clinical susceptibility to WNV infection. Similar studies in horses, a particularly susceptible species, have been lacking, in part, because of the difficulty in collecting populations sufficiently homogenous in their infection and disease states. The equine OAS gene cluster most closely resembles the human cluster, with single copies of OAS1, OAS3 and OAS2 in the same orientation. With naturally occurring susceptible and resistant sub-populations to lethal West Nile encephalitis, we undertook a case-control association study to investigate whether, similar to humans (OAS1) and mice (Oas1b), equine OAS1 plays a role in resistance to severe WNV infection. We identified naturally occurring single nucleotide mutations in equine (Equus caballus) OAS1 and RNASEL genes and, using Fisher's Exact test, we provide evidence that mutations in equine OAS1 contribute to host susceptibility. Virtually all of the associated OAS1 polymorphisms were located within the interferon-inducible promoter, suggesting that differences in OAS1 gene expression may determine the host's ability to resist clinical manifestations associated with WNV infection.