Multidrug-Resistant ESBL/AmpC-Producing Klebsiella pneumoniae Isolated from Healthy Thoroughbred Racehorses in Japan

Abstract

Extended-spectrum β-lactamase (ESBL)- and AmpC β-lactamase (AmpC)-producing Klebsiella spp. have become a major health problem, leading to treatment failure in humans and animals. This study aimed to evaluate the presence of ESBL/AmpC-producing Klebsiella spp. isolated from racehorses in Japan. Feces samples from 212 healthy Thoroughbred racehorses were collected from the Japan Racing Association Training Centers between March 2017 and August 2018. ESBL/AmpC-producing Klebsiella spp. were isolated using selective medium containing 1 µg/mL cefotaxime. All isolates were subjected to bacterial species identification (MALDI-TOF MS), antimicrobial susceptibility test (disk diffusion test), characterization of resistance genes (PCR), conjugation assay, and genetic relatedness (multilocus sequence typing/MLST). Twelve ESBL/AmpC-producing Klebsiella pneumoniae (ESBL/AmpC-KP) were isolated from 3.3% of horse samples. Antimicrobial resistance profiling for 17 antimicrobials showed all ESBL/AmpC-KP were multidrug-resistant (MDR). Only 1 isolate was confirmed as an ESBL producer (bla_CTX-M-2-positive), whereas the other 11 isolates were plasmid-mediated AmpC (pAmpC) producers (bla_CMY positive). On the basis of MLST analysis, the ESBL-KP isolate was identified as sequence type (ST)-133 and four different STs among AmpC-KP isolates, ST-145, ST-4830, ST-4831, and ST-4832, were found to share six of the seven loci constituting a single-locus variant. This is the first study to show K. pneumoniae carrying MDR pAmpC isolated from a racehorse.

Keywords: AmpC β-lactamase; Klebsiella pneumoniae; extended-spectrum β-lactamase; horse; multidrug resistance.

Figure 1

Characteristics of extended-spectrum β-lactamase/AmpC β-lactamase-producing Klebsiella pneumoniae (ESBL/AmpC-KP) isolated from Thoroughbred racehorses in Japan. ¹ Miho, Miho Training Center, Japan Racing Association, Ibaraki; Ritto, Ritto Training Center, Japan Racing Association, Shiga. ² ABPC, ampicillin; CXM, cefuroxime; CTX, cefotaxime; CAZ, ceftazidime; GM, gentamicin; KM, kanamycin; SM, streptomycin; TC, tetracycline; OTC, oxytetracycline; DOXY, doxycycline; CP, chloramphenicol; CL, colistin; NA, nalidixic acid; NFLX, norfloxacin; MAR, marbofloxacin; FOM, fosfomycin; STX, trimethoprim/sulfamethoxazole; red, resistance; yellow, intermediate; green, susceptible.