Department of Microbiology, University of Agriculture in Cracow, Mickiewicza Ave 24/28, 30-059, Cracow, Poland.
Environ Sci Pollut Res Int. 2018 Aug;25(22):21789-21800. doi: 10.1007/s11356-018-2274-x. Epub 2018 May 23.
The aim of the study was to determine the antimicrobial resistance profile and the occurrence of extended-spectrum beta-lactamase genes and to analyze the genetic diversity of Escherichia coli strains isolated from the environment of horse riding centers. The study was conducted using E. coli strains isolated from the air, manure, and horse nostril swabs in three horse riding centers differing in the system of horse keeping-stable (OJK Pegaz and KJK Szary) and free-range (SKH Nielepice). Resistance to antibiotics was determined using the disk-diffusion method, and the PCR technique was employed to detect the extended-spectrum β-lactamase (ESBL) genes, while the genetic diversity of strains was assessed by rep-PCR. A total of 200 strains were collected during the 2-year study, with the majority isolated from KJK Szary, while the smallest number was obtained from SKH Nielepice. The strains were mostly resistant to ampicillin, aztreonam, and ticarcillin. The tested strains were most frequently resistant to one or two antibiotics, with a maximum of ten antimicrobials at the same time. Two multidrug-resistant (MDR) strains were detected in OJK Pegaz while in KJK Szary there were two MDR and one extensively drug-resistant (XDR) strain. The ESBL mechanism was most frequently observed in OJK Pegaz (20.31% of strains) followed by KJK Szary (15.53% of strains) and SKH Nielepice (15.15% of strains). Among the ESBL-determining genes, only blaTEM and blaCTXM-9 were detected-blaTEM was mostly found in KJK Szary (53.40% of strains), while the second detected gene-blaCTXM-9-was most frequent in SKH Nielepice (6.06% of strains). The rep-PCR genotyping showed high variation among the analyzed strains, whereas its degree differed between the studied facilities, indicating that the type of horse keeping (stable vs. free-range) affects the genetic diversity of the E. coli strains. Having regard to the fact that the tested strains of E. coli were derived from non-hospitalized horses that were not treated pharmacologically, we can assume that the observed antimicrobial resistance may be of both-natural origin, i.e., not the result of the selection pressure, and acquired, the source of which could be people present in the horse riding facilities, the remaining horses which were not included in the study, and air, as well as water, fodder, and litter of the animals. Therefore, it can be concluded that the studied horses are the source of resistant E. coli and it is reasonable to continue monitoring the changes in antimicrobial resistance in those bacteria.
本研究旨在确定从马厩环境中分离的大肠杆菌菌株的抗菌药物耐药谱、产超广谱β-内酰胺酶基因的情况,并分析其遗传多样性。研究采用空气、粪便和马鼻腔拭子中分离的大肠杆菌菌株,在系统上分为三种不同的马厩:封闭式(OJK Pegaz 和 KJK Szary)和散养式(SKH Nielepice)。使用纸片扩散法测定抗生素的耐药性,采用 PCR 技术检测产超广谱β-内酰胺酶(ESBL)基因,采用重复片段聚合酶链反应(rep-PCR)评估菌株的遗传多样性。在为期两年的研究中,共收集了 200 株菌株,其中大部分分离自 KJK Szary,而来自 SKH Nielepice 的菌株数量最少。这些菌株对氨苄西林、氨曲南和替卡西林的耐药率最高。受试菌株主要对一种或两种抗生素耐药,同时对最多十种抗菌药物耐药。在 OJK Pegaz 中检测到两株多药耐药(MDR)菌株,在 KJK Szary 中检测到两株 MDR 和一株广泛耐药(XDR)菌株。在 ESBL 机制中,OJK Pegaz 中观察到的比例最高(20.31%的菌株),其次是 KJK Szary(15.53%的菌株)和 SKH Nielepice(15.15%的菌株)。在 ESBL 决定基因中,只检测到 blaTEM 和 blaCTXM-9,blaTEM 主要在 KJK Szary 中发现(53.40%的菌株),而检测到的第二种基因 blaCTXM-9 在 SKH Nielepice 中最为常见(6.06%的菌株)。重复片段聚合酶链反应基因分型显示,分析的菌株之间存在高度的变异性,而在不同的研究设施之间,其程度存在差异,表明马的饲养方式(封闭式与散养式)会影响大肠杆菌菌株的遗传多样性。鉴于受试的大肠杆菌菌株来源于未接受药物治疗的非住院马,我们可以假设观察到的抗菌药物耐药性可能既有天然来源(即非选择压力所致),也有后天获得性来源,其来源可能是人在马厩设施中,未包括在研究中的其余马匹,以及空气、水、饲料和动物的垫料。因此,可以得出结论,研究中的马匹是耐药大肠杆菌的来源,有理由继续监测这些细菌中抗菌药物耐药性的变化。