Prevalence of Microbial Isolates Cultured from Endometrial Swab Samples Collected from United Kingdom Thoroughbred Mares from 2014 to 2020.

Determining whether endometrial microbial isolates are pathogens, contaminants, or even part of the "normal" microbiome is extremely complex, particularly given the absence of "gold standard" tests for endometritis. Population-level benchmarking and temporal monitoring can provide novel insights and a wider context to improve understanding. This study aimed to (i) estimate the prevalence of endometrial isolates from swabs of Thoroughbred broodmares in Newmarket, UK between 2014 and 2020; and (ii) evaluate the effects of year, mare age, and cytology findings on isolate prevalence. Generalised linear mixed models with a logit link, both null models and models using year of sampling, mare age, or cytology findings as predictors, were fitted to estimate isolate prevalence. Over the 7-year period, data were available from 18,996 endometrial-swab samples from 6050 mares on 290 premises. The overall isolate prevalence was 35.5% (95% confidence interval (CI) 33.0-37.9), and this varied significantly between years. The most prevalent isolates were ß-hemolytic Streptococcus (17.9; 95% CI: 17-19) and E. coli (10.3%; 95% CI: 9.0-11.6). Isolate prevalence increased with mare age except for E. coli isolates, and with increasing category of cytology findings except for α-hemolytic Streptococcus isolates. The results provide novel estimates of isolate prevalence and highlight knowledge gaps around potential complexities in the interpretation of findings.

Antimicrobial resistance of endometrial bacterial isolates collected from UK Thoroughbred mares between 2014 and 2020.

BACKGROUND: Antimicrobial resistance (AMR) is important in equine reproduction, as antimicrobials have historically been widely used in the management of breeding mares. However, evidence of the characteristics of AMR in uterine isolates is limited in the UK. The objective of this retrospective study was therefore to describe temporal changes in AMR patterns of bacteria isolated from the endometrium of Thoroughbred broodmares in south-east England between 2014 and 2020. METHOD: Endometrial swabs were processed for microbiology and antimicrobial susceptibility testing (AST). For frequently isolated bacteria, changes in AMR patterns over time were assessed using a logistic regression model. RESULTS: From 18,996 endometrial swabs, 30.5% were positive for microbial culture. AST was performed on 2091 isolates, representing 1924 swabs collected from 1370 mares located at 132 premises. Beta-haemolytic Streptococcus (BHS, 52.5%) and Escherichia coli (25.8%) were most frequently isolated. In BHS, resistance to enrofloxacin (p = 0.02), nitrofurazone (p < 0.001) and oxytetracycline (p < 0.01) increased significantly between 2014 and 2020, while resistance to trimethoprim-sulfamethoxazole (p < 0.001) decreased. In E. coli, resistance to nitrofurazone increased (p = 0.04) and resistance to gentamycin (p = 0.02) and trimethoprim-sulfamethoxazole (p < 0.001) decreased. LIMITATIONS: Variations in the specimen collection protocols might have affected the frequency of isolates detected. CONCLUSION: Between 2014 and 2020, AMR changed in this bacterial population. However, there was no significant increase in resistance to penicillin (99.6% BHS susceptible), gentamycin (81.7% E. coli susceptible) or ceftiofur.

Bayesian accuracy estimates and fit for purpose thresholds of cytology and culture of endometrial swab samples for detecting endometritis in mares.

The overall aim of this work was to identify the potential impact of misclassification errors associated with routine screening and diagnostic testing for endometritis in mares. Using Bayesian latent class models (BLCM), specific objectives were to: 1) estimate the diagnostic accuracy of cytology and culture of endometrial swab samples to detect endometritis in mares; 2) assess the impact of different cytology thresholds on test accuracy and misclassification costs; and 3) assess the sensitivity (Se) and specificity (Sp) of a diagnostic strategy including both tests interpreted in series and parallel. Diagnostic and pre-breeding endometrial swab samples collected from 3448 mares based at breeding premises located in the South East of England between 2014 and 2020 were retrospectively analysed. Culture results were classified as positive according to three different case definitions: (A) > 90% of the growth colonies were a monoculture; (B) pathogenic or pathogenic and non-pathogenic bacteria were identified; and (C) any growth was observed. Endometrial smears were graded based on the percent of polymorphonuclear cells (PMN) per high power field (HPF). A hierarchical BLCM was fitted using the cross-tabulated results of the three culture case definitions with a cytology threshold fixed at > 0.5% PMN. Fit for purpose cytology thresholds were proposed using a misclassification cost analysis in the context of good antimicrobial stewardship and for varying endometritis prevalence estimates. Median [95% Bayesian credible intervals (BCI)] cytology Se estimates were 6.5% (2.2-11.6), 6.4% (2.2-10.8) and 6.3% (2.2-10.8) for scenario A, B and C, respectively. Median (95% BCI) cytology Sp estimates were 88.8% (83.1-94.8), 88.9% (83.9-93.8) and 88.8% (84.0-93.8) for scenarios A, B and C, respectively. Median (95% BCI) culture Se estimates were 37.5% (29.9-46.0), 42.3% (33.8-51.1) and 46.4% (35.7-55.9) for scenarios A, B and C, respectively. Median (95% BCI) culture Sp estimates were 92.8% (84.3-99.0), 91.5% (82.5-98.0) and 90.8% (80.1-97.4) for scenarios A, B and C, respectively. Regardless of the culture case definition, Se and Sp of cytology (> 0.5% PMN) was lower than previously reported for swab samples in studies using histology as the reference standard test. The misclassification cost term decreased as the cytology threshold increased for all scenarios and all prevalence contexts, suggesting that, regardless of the endometritis prevalence in the population, increasing the cytology threshold would reduce the misclassification costs associated with false positive mares contributing to good antimicrobial stewardship.

The Effect of Routine Postpartum Uterine Lavage on Endometrial Cytology, Culture, and Pregnancy Rates in Thoroughbred Broodmares.

Minimizing the interval between parturition and successful rebreeding of thoroughbred broodmares is important in maintaining annual foal production and broodmare profitability. Routine postpartum uterine lavage may help achieve this by aiding uterine clearance and involution in the early postpartum period and increasing pregnancy rates. This retrospective study investigates the effect of postpartum uterine lavage in 32 thoroughbred mares housed on a single farm in Australia in 2014 and 2015. In the control year (2014), postpartum uterine lavage was not performed. In the treatment year (2015), all mares received a uterine lavage 3 days postpartum. In both years, endometrial swabs were obtained for culture and cytology at the first postpartum estrus (foal heat swab) and the second postpartum estrus (second heat swab), prior to the mare being bred by natural cover at the second postpartum estrus. There was no significant difference in bacterial culture rates from foal heat or second heat swabs between the control and treatment years. Assessment of endometrial cytology of the foal heat swab and second heat swab revealed a significantly higher proportion of polymorphonuclear cells (PMNs) in the control year compared with the treatment year. Positive culture and cytology rates were significantly higher at foal heat than at the second postpartum heat in both years. No association of year, cytology, or culture with pregnancy rates was identified. In conclusion, routine postpartum uterine lavage significantly reduces the presence of endometrial PMNs at first and second postpartum estrus but does not reduce endometrial culture rates or increase pregnancy rates.