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智利鲑鱼养殖场分离出的细菌对氟甲喹敏感性降低的机制特征分析

Characterization of Mechanisms Lowering Susceptibility to Flumequine among Bacteria Isolated from Chilean Salmonid Farms.

作者信息

Concha Christopher, Miranda Claudio D, Hurtado Luz, Romero Jaime

机构信息

Laboratorio de Tecnología Enzimática para Bioprocesos (TEB), Universidad de La Serena, La Serena 1700000, Chile.

Laboratorio de Silvigenómica y Biotecnología (SILGENBIO), Universidad de La Serena, La Serena 1700000, Chile.

出版信息

Microorganisms. 2019 Dec 14;7(12):698. doi: 10.3390/microorganisms7120698.

Abstract

Despite their great importance for human therapy, quinolones are still used in Chilean salmon farming, with flumequine and oxolinic acid currently approved for use in this industry. The aim of this study was to improve our knowledge of the mechanisms conferring low susceptibility or resistance to quinolones among bacteria recovered from Chilean salmon farms. Sixty-five isolates exhibiting resistance, reduced susceptibility, or susceptibility to flumequine recovered from salmon farms were identified by their 16S rRNA genes, detecting a high predominance of species belonging to the genus (52%). The minimum inhibitory concentrations (MIC) of flumequine in the absence and presence of the efflux pump inhibitor (EPI) Phe-Arg-β-naphthylamide and resistance patterns of isolates were determined by a microdilution broth and disk diffusion assays, respectively, observing MIC values ranging from 0.25 to >64 µg/mL and a high level of multi-resistance (96%), mostly showing resistance to florfenicol and oxytetracycline. Furthermore, mechanisms conferring low susceptibility to quinolones mediated by efflux pump activity, quinolone target mutations, or horizontally acquired resistance genes (, , , ) were investigated. Among isolates exhibiting resistance to flumequine (≥16 µg/mL), the occurrence of chromosomal mutations in target protein GyrA appears to be unusual (three out of 15), contrasting with the high incidence of mutations in GyrB (14 out of 17). Bacterial isolates showing resistance or reduced susceptibility to quinolones mediated by efflux pumps appear to be highly prevalent (49 isolates, 75%), thus suggesting a major role of intrinsic resistance mediated by active efflux.

摘要

尽管喹诺酮类药物对人类治疗非常重要,但它们仍在智利鲑鱼养殖中使用,目前氟甲喹和恶喹酸已被批准用于该行业。本研究的目的是增进我们对从智利鲑鱼养殖场分离出的细菌中赋予对喹诺酮类药物低敏感性或耐药性机制的了解。通过16S rRNA基因对从鲑鱼养殖场分离出的65株对氟甲喹表现出耐药性、敏感性降低或敏感的菌株进行鉴定,检测到属于该属的物种占主导地位(52%)。分别通过微量稀释肉汤法和纸片扩散法测定了在不存在和存在外排泵抑制剂(EPI)苯丙氨酸 - 精氨酸 - β - 萘酰胺的情况下氟甲喹的最低抑菌浓度(MIC)以及菌株的耐药模式,观察到MIC值范围为0.25至>64μg/mL,并且多耐药水平较高(96%),大多对氟苯尼考和土霉素耐药。此外,还研究了由外排泵活性、喹诺酮类药物靶点突变或水平获得的耐药基因(、、、)介导的对喹诺酮类药物低敏感性的机制。在对氟甲喹耐药(≥16μg/mL)的菌株中,靶蛋白GyrA发生染色体突变的情况似乎不常见(15株中有3株),这与GyrB中突变的高发生率形成对比(17株中有14株)。由外排泵介导对喹诺酮类药物耐药或敏感性降低的细菌分离株似乎非常普遍(49株,75%),因此表明主动外排介导的固有耐药起主要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b96/6955667/9d216fd52286/microorganisms-07-00698-g001.jpg

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