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首页> 外文期刊>Applied Microbiology >Strain Variability of Listeria monocytogenes under NaCl Stress Elucidated by a High-Throughput Microbial Growth Data Assembly and Analysis Protocol
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Strain Variability of Listeria monocytogenes under NaCl Stress Elucidated by a High-Throughput Microbial Growth Data Assembly and Analysis Protocol

机译：通过高通量微生物生长数据组装和分析方案阐明李斯特菌李斯特菌单核细胞增生的菌株变异性

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Listeria monocytogenes causes the severe foodborne illness listeriosis and survives in food-associated environments due to its high stress tolerance. A data assembly and analysis protocol for microbial growth experiments was compiled to elucidate the strain variability of L. monocytogenes stress tolerance. The protocol includes measurement of growth ability under stress (step 1), selection of a suitable method for growth parameter calculation (step 2), comparison of growth patterns between strains (step 3), and biological interpretation of the discovered differences (step 4). In step 1, L. monocytogenes strains ( n ?=?388) of various serovars and origins grown on media with 9.0% NaCl were measured using a Bioscreen C microbiology reader. Technical variability of the growth measurements was assessed and eliminated. In step 2, the growth parameters determined by Gompertz, modified-Gompertz, logistic, and Richards models and model-free splines were compared, illustrating differences in the suitability of these methods to describe the experimental data. In step 3, hierarchical clustering was used to describe the NaCl tolerance of L. monocytogenes measured by strain-specific variation in growth ability; tolerant strains had higher growth rates and maximum optical densities and shorter lag phases than susceptible strains. The spline parameter area under the curve best classified “poor,” “average,” and “good” growers. In step 4, the tested L. monocytogenes lineage I strains (serovars 4b and 1/2b) proved to be significantly more tolerant toward 9.0% NaCl than lineage II strains (serovars 1/2a, 1/2c, and 3a). Our protocol provides systematic tools to gain comparable data for investigating strain-specific variation of bacterial growth under stress.IMPORTANCE The pathogen Listeria monocytogenes causes the foodborne disease listeriosis, which can be fatal in immunocompromised individuals. L. monocytogenes tolerates several environmental stressors and can persist in food-processing environments and grow in foodstuffs despite traditional control measures such as high salt content. Nonetheless, L. monocytogenes strains differ in their ability to withstand stressors. Elucidating the intraspecies strain variability of L. monocytogenes stress tolerance is crucial for the identification of particularly tolerant strains. To enhance reliable identification of variability in bacterial stress tolerance phenotypes, we compiled a large-scale protocol for the entire data assembly and analysis of microbial growth experiments, providing a systematic approach and checklist for experiments on strain-specific growth ability. Our study illustrated the diversity and strain-specific variation of L. monocytogenes stress tolerance with an unprecedented scope and discovered biologically relevant serovar- and lineage-dependent phenotypes of NaCl tolerance.
机译：由于其高应力耐受性，Listeria单核细胞增强引起严重的食物中患病局部障碍，并在食物相关环境中存活。编制了用于微生物生长实验的数据组件和分析方案，以阐明L.单核细胞生成胁迫耐受性的应变变异性。该方案包括在应力下的生长能力的测量（步骤1），选择合适的生长参数计算方法（步骤2），菌株之间的生长模式（步骤3）和发现的差异的生物解释的比较（步骤4） 。在步骤1中，使用生物筛C微生物学读者测量在培养基上生长的各种血细胞增生的单核细胞增生（N？=β388）和在培养基上生长的培养基。评估和消除了生长测量的技术可变性。在步骤2中，比较了由Gompertz，修改的Gompertz，逻辑和理查兹模型确定的生长参数和无模型样条曲线，说明这些方法描述实验数据的适用性的差异。在步骤3中，使用分层聚类来描述通过生长能力的应变特异性变异测量的L.单核细胞增生的NaCl耐受性;耐受性速率较高的生长速率和最大光学密度和比易感菌株更短的滞后阶段。花键参数面积在曲线下最佳分类“差”，“平均”和“好”种植者。在步骤4中，证明了测试的L.单核细胞增生谱系I株（Serovars 4b和1 / 2b）比谱系II株（Serovars 1 / 2a，1 / 2c和3a）明显更高耐受9.0％NaCl。我们的协议提供了系统的工具，以获得用于研究胁迫下细菌生长的应变特异性变异的可比数据。分析病原体李斯特菌，使食物造成疾病局部病变引起，这可能是免疫功能性的致命性。 L.单核细胞增生耐受几种环境压力源，尽管传统的控制措施如高盐含量，但仍可持续存在食品加工环境并在食品中生长。尽管如此，L.单核细胞增生菌株的抵抗能力不同。阐明L.单核细胞增强胁迫耐受性的缠结菌株可变性对于鉴定特别耐受性菌株至关重要。为了增强细菌胁迫性耐受性表型的可变性鉴定，我们编制了一个大规模的方案，用于整个数据组件和微生物生长实验分析，提供系统的方法和清单，用于实验菌株特异性生长能力。我们的研究说明了L.单核细胞增强胁迫耐受性的多样性和菌株特异性变异，并发现了NaCl耐受的生物相关的血清和依赖性表型。

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