Abstract

This study investigated the aminoglycoside resistance phenotypes and genotypes, as well as the prevalence of virulence genes, in Enterococcus species isolated from clinical patients in China. A total of 160 enterococcal isolates from various clinical samples collected from September 2013 to July 2014 were identified to the species level using the VITEK-2 COMPACT system. The antimicrobial susceptibilities of the identified Enterococcus strains were determined by the Kirby-Bauer (K-B) disc diffusion method. PCR-based assays were used to detect the aminoglycoside resistance and virulence genes in all enterococcal isolates. Of 160 Enterococcus isolates, 105 were identified as E. faecium, 35 as E. faecalis, and 20 isolates were classified as “other” Enterococcus species. High-level aminoglycoside resistance (HLAR) for gentamicin, streptomycin, and both antibiotics was identified in 58.8, 50, and 34.4% of strains, respectively. The most common virulence gene (50.6% of isolates) was efaA, followed by asa1 (28.8%). The most prevalent aminoglycoside resistance genes were aac(6')-Ie-aph(2''), aph(2')-Id, aph(3')-IIIa, and ant(6')-Ia, present in 49.4%, 1.3%, 48.8% and 31.3% of strains, respectively. Overall, E. faecium and E. faecalis were most frequently associated with hospital-acquired enterococcal infections in Zhejiang Province. All aminoglycoside resistance genes, except aph(2'')-Id, were significantly more prevalent in HLAR strains than amongst high level aminoglycoside susceptible (HLAS) strains, while there was no significant difference between HLAR and HLAS strains in regard to the prevalence of virulence genes, apart from esp, therefore, measures should be taken to manage infections caused by multi-drug resistant Enterococcus species.

Highlights

  • Enterococcus species are opportunistic pathogens that are part of the normal gut microbiota of both humans and animals, and can survive in a diverse range of harsh conditions [1,2]

  • Amongst the high-level aminoglycoside-resistant (HLAR) strains, comparison of the two species showed that rates of resistance were significantly higher in E. faecium than in E. faecalis for all antibiotics examined, except ERY, LZD, and CHL

  • Comparison of HLAR vs. HLAS E. faecium isolates revealed that rates of antibiotic resistance were only significantly different for TEC and VAN, while a significant difference was only observed for erythromycin between the HLAR and HLAS

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Summary

Introduction

Enterococcus species are opportunistic pathogens that are part of the normal gut microbiota of both humans and animals, and can survive in a diverse range of harsh conditions [1,2]. Enterococcus species to be identified was isolated from an endocarditis patient in the 1900s. Enterococci have been recognized as an important cause of nosocomial infections [3]. High-level aminoglycoside-resistant (HLAR) enterococci and vancomycin-resistant enterococci (VRE) have caused significant problems for clinical anti-infective therapy. HLAR strains are becoming more prevalent, mostly because of aminoglycoside modifying enzymes (AMEs) that are encoded within mobile genetic elements, which are widespread amongst enterococci [5]. Among the AMEs, the most prevalent gene is aac(6')-Ie-aph(2''), which encodes a bifunctional enzyme, AAC(6')-APH(2''), that confers resistance to a broad spectrum of aminoglycosides [6]. Other resistance genes, encoding the AMEs 2'-O-phosphotransferase (APH(2')), 3'-O-phosphotransferase (APH(3')), 3'-O-adenyltransferase (ANT(3')), 4-O-adenyltransferase (ANT(4')), and 6'-O-adenyltransferase (ANT(6')) are found on these mobile genetic elements. Enterococci produce many virulence factors, including collagen-binding protein (ace), aggregation substance (asa1), cytolysin (cylA), endocarditis antigen (efaA), enterococcal surface protein (esp), gelatinase (gelE), and hyaluronidase (hyl) [7,8,9]

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