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从肉类加工设施分离出的单核细胞增生李斯特菌菌株对苯扎氯铵的耐受性与质粒携带的bcrABC操纵子的存在有关。

Benzalkonium chloride tolerance of Listeria monocytogenes strains isolated from a meat processing facility is related to presence of plasmid-borne bcrABC cassette.

作者信息

Minarovičová Jana, Véghová Adriana, Mikulášová Mária, Chovanová Romana, Šoltýs Katarína, Drahovská Hana, Kaclíková Eva

机构信息

Food Research Institute, National Agricultural and Food Centre, Priemyselná 4, 82475, Bratislava, Slovakia.

Department of Molecular Biology, Faculty of Natural Sciences, Comenius University, Bratislava, Slovakia.

出版信息

Antonie Van Leeuwenhoek. 2018 Oct;111(10):1913-1923. doi: 10.1007/s10482-018-1082-0. Epub 2018 Apr 6.

Abstract

Listeria monocytogenes is a serious foodborne pathogen capable of persisting in food processing environments. Tolerance to disinfectants used in industrial settings constitutes an important factor of Listeria survival. In the present study, the mechanism of tolerance to benzalkonium chloride (BAC) was investigated in 77 L. monocytogenes isolates from a meat facility. By PCR approach, the mdrL and lde chromosomal efflux pump genes were detected in all isolates. No isolate was positive for qacH and emrE genes. However, the bcrABC cassette was present in 17 isolates of serogroup IIa possessing the same AscI/ApaI pulsotype, the operon being localized on a plasmid. The significant relation of BAC tolerance with bcrABC presence was confirmed as all bcrABC positive isolates showed the highest minimal inhibitory concentration (MIC) values for BAC and increased sensitivity to BAC was observed after plasmid curing. No effect of the efflux pump inhibitor reserpine on BAC tolerance in bcrABC positive strains was observed in contrast to all bcrABC negative strains. Lower ethidium bromide efflux in bcrABC positive isolates compared to bcrABC negative and plasmid-cured L. monocytogenes isolates was observed. The expression of bcrABC genes was BAC-induced. The confirmed effect of bcrABC to increased BAC tolerance, coupled with its plasmid location, may be an important factor in potential dissemination of the biocide resistance among Listeria species. The understanding of molecular mechanisms of biocide tolerance should help to improve control measures to prevent further spread of L. monocytogenes in food production environments with frequent use of BAC.

摘要

单核细胞增生李斯特菌是一种严重的食源性病原体,能够在食品加工环境中持续存在。对工业环境中使用的消毒剂的耐受性是李斯特菌生存的一个重要因素。在本研究中,对来自一家肉类加工厂的77株单核细胞增生李斯特菌分离株对苯扎氯铵(BAC)的耐受机制进行了研究。通过PCR方法,在所有分离株中均检测到mdrL和lde染色体外排泵基因。没有分离株对qacH和emrE基因呈阳性。然而,bcrABC操纵子存在于17株血清型IIa的分离株中,这些分离株具有相同的AscI/ApaI脉冲型,该操纵子位于一个质粒上。BAC耐受性与bcrABC的存在之间的显著关系得到了证实,因为所有bcrABC阳性分离株对BAC的最低抑菌浓度(MIC)值最高,并且在质粒消除后观察到对BAC的敏感性增加。与所有bcrABC阴性菌株相比,未观察到外排泵抑制剂利血平对bcrABC阳性菌株的BAC耐受性有影响。与bcrABC阴性和质粒消除的单核细胞增生李斯特菌分离株相比,观察到bcrABC阳性分离株中溴化乙锭的外排较低。bcrABC基因的表达是由BAC诱导的。bcrABC对增加BAC耐受性的已证实作用,再加上其质粒定位,可能是李斯特菌属中生物杀灭剂抗性潜在传播的一个重要因素。了解生物杀灭剂耐受性的分子机制应有助于改进控制措施,以防止单核细胞增生李斯特菌在频繁使用BAC的食品生产环境中进一步传播。

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