Extended-Spectrum β-lactamase-Producing Enterobacteriaceae Shedding in Farm Horses Versus Hospitalized Horses: Prevalence and Risk Factors

SIMPLE SUMMARY: This prospective study investigated the prevalence, molecular characteristics and risk factors of extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae (ESBL-E) shedding in three equine cohorts: (i) farm horses (13 farms, n = 192); (ii) on admission to a hospital (n = 168) and; (iii) horses hospitalized for ≥72 h re-sampled from cohort (ii) (n = 86). Bacteria were isolated from rectal swabs, identified, antibiotic susceptibility patterns were determined, and medical records and owners’ questionnaires were analyzed for risk factor analysis. ESBL shedding rates significantly increased during hospitalization (77.9%, n = 67/86), compared to farms (20.8%, n = 40/192), and horses on admission (19.6%, n = 33/168). High bacterial species diversity was identified, mainly in cohorts (ii) and (iii), with high resistance rates to commonly used antimicrobials. Risk factors for shedding in farms included horses’ breed (Arabian), sex (stallion), and antibiotic treatment. Older age was identified as a protective factor. We demonstrated a reservoir for antibiotic-resistant bacteria in an equine hospital and farms, with a significant ESBL-E acquisition. In light of our findings, in order to control ESBL spread, we recommend conducting active ESBL surveillance programs alongside antibiotic stewardship programs in equine facilities. ABSTRACT: We aimed to investigate the prevalence, molecular characteristics and risk factors of extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae (ESBL-E) shedding in horses. A prospective study included three cohorts: (i) farm horses (13 farms, n = 192); (ii) on hospital admission (n = 168) and; (iii) horses hospitalized for ≥72 h re-sampled from cohort (ii) (n = 86). Enriched rectal swabs were plated, ESBL-production was confirmed (Clinical and Laboratory Standards Institute (CLSI)) and genes were identified (polymerase chain reaction (PCR)). Identification and antibiotic susceptibility were determined (Vitek-2). Medical records and owners’ questionnaires were analyzed. Shedding rates increased from 19.6% (n = 33/168) on admission to 77.9% (n = 67/86) during hospitalization (p < 0.0001, odds ratio (OR) = 12.12). Shedding rate in farms was 20.8% (n = 40/192), significantly lower compared to hospitalized horses (p < 0.0001). The main ESBL-E species (n = 192 isolates) were E. coli (59.9%, 115/192), Enterobacter sp. (17.7%, 34/192) and Klebsiella pneumoniae (13.0%, 25/192). The main gene group was CTX-M-1 (56.8%). A significant increase in resistance rates to chloramphenicol, enrofloxacin, gentamicin, nitrofurantoin, and trimethoprim-sulpha was identified during hospitalization. Risk factors for shedding in farms included breed (Arabian, OR = 3.9), sex (stallion, OR = 3.4), and antibiotic treatment (OR = 9.8). Older age was identified as a protective factor (OR = 0.88). We demonstrated an ESBL-E reservoir in equine cohorts, with a significant ESBL-E acquisition, which increases the necessity to implement active surveillance and antibiotic stewardship programs.