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Biocide tolerance, phenotypic and molecular response of Lactic Acid Bacteria isolated from naturally-fermented Aloreña table olives throughout fermentation to induction by different physico-chemical stresses

机译:从自然发酵的Aloreña食用橄榄中分离出的乳酸菌在整个发酵过程中的杀菌剂耐受性,表型和分子响应,以不同的理化胁迫诱导

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摘要

Lactic acid bacteria isolated from Aloreña table olives throughout fermentation process were resistant at least to three antibiotics (Casado Muñoz et al., 2014), however they were very sensitive to all biocides tested in this study (MIC below the epidemiological cut-off values “ECOFF” determined in the present study) except 2-15% of Lc. pseudomesenteroides which were resistant to hexachlorophene and Lb. pentosus to cetrimide and hexadecylpiridinium. To give new insights of how LAB become resistant in a changing environment, the effect of different physico-chemical stresses -including antimicrobials- on phenotypic and genotypic responses of LAB was analyzed in the present study. The results obtained indicated that a similar phenotypic response was obtained under all stress conditions tested (antimicrobials, chemicals and UV light) producing changes in susceptibility patterns of antibiotics (increased MICs for ampicillin, chloramphenicol, ciprofloxacin, teicoplanin and tetracycline, while decreased MICs were shown for clindamycin, erythromycin, streptomycin and trimethoprim in the majority of strains). By means of statistical analysis, cross resistance between different antibiotics was detected in all stress conditions. However, expression profiles of selected genes involved in stress/resistance (rpsL, recA, uvrB and srtA genes) were different depending on the stress parameter, LAB species and strain, and also the target gene. We can conclude that, in spite of the uniform phenotypic responses to several stresses, the repertoire of induced and repressed genes were different upon the stress parameter and the LAB strain. So, a search for a target to improve stress tolerance of LAB especially those of importance as starter/protective cultures or as probiotics may depend on the individual screening of each strain although, we could predict the antibiotic phenotypic response to all stresses.
机译:在整个发酵过程中,从Aloreña食用橄榄分离出的乳酸菌至少对三种抗生素具有抗药性(CasadoMuñoz等,2014),但是它们对本研究中测试的所有生物杀灭剂都非常敏感(MIC低于流行病学临界值以下)本研究中确定的“ ECOFF”除外),其中Lc的含量为2-15%。对六氯苯和Lb有抗性的假美沙龙。戊糖到西曲美和十六烷基哌啶鎓。为了提供关于LAB如何在变化的环境中具有抗性的新见解,在本研究中分析了不同的物理化学应激(包括抗微生物剂)对LAB的表型和基因型反应的影响。获得的结果表明,在所有测试的胁迫条件下(抗微生物剂,化学药品和紫外线)都获得了相似的表型反应,从而改变了抗生素的敏感性模式(氨苄青霉素,氯霉素,环丙沙星,替考拉宁和四环素的MIC升高),而MIC却降低了。在大多数菌株中用于克林霉素,红霉素,链霉素和甲氧苄啶)。通过统计分析,在所有应激条件下均检测到不同抗生素之间的交叉耐药性。但是,取决于胁迫参数,LAB种类和菌株以及目标基因,参与胁迫/抗性的选定基因(rpsL,recA,uvrB和srtA基因)的表达谱是不同的。我们可以得出结论,尽管对几个应力有统一的表型响应,但在应力参数和LAB菌株上,诱导和抑制基因的组成是不同的。因此,尽管我们可以预测对所有压力的抗生素表型反应,但寻找能够提高LAB压力耐受性的目标,尤其是作为起子/保护性培养物或益生菌的目标,可能取决于每个菌株的个体筛选。

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