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首页> 外文期刊>Applied and Environmental Microbiology >Molecular Fingerprinting of Dairy Microbial Ecosystems by Use of Temporal Temperature and Denaturing Gradient Gel Electrophoresis
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Molecular Fingerprinting of Dairy Microbial Ecosystems by Use of Temporal Temperature and Denaturing Gradient Gel Electrophoresis

机译：利用温度和变性梯度凝胶电泳对乳制品微生物生态系统进行分子指纹分析

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Numerous microorganisms, including bacteria, yeasts, and molds, constitute the complex ecosystem present in milk and fermented dairy products. Our aim was to describe the bacterial ecosystem of various cheeses that differ by production technology and therefore by their bacterial content. For this purpose, we developed a rapid, semisystematic approach based on genetic profiling by temporal temperature gradient electrophoresis (TTGE) for bacteria with low-G+C-content genomes and denaturing gradient gel electrophoresis (DGGE) for those with medium- and high-G+C-content genomes. Bacteria in the unknown ecosystems were assigned an identity by comparison with a comprehensive bacterial reference database of ～150 species that included useful dairy microorganisms (lactic acid bacteria), spoilage bacteria (e.g., Pseudomonas and Enterobacteriaceae), and pathogenic bacteria (e.g., Listeria monocytogenes and Staphylococcus aureus). Our analyses provide a high resolution of bacteria comprising the ecosystems of different commercial cheeses and identify species that could not be discerned by conventional methods; at least two species, belonging to the Halomonas and Pseudoalteromonas genera, are identified for the first time in a dairy ecosystem. Our analyses also reveal a surprising difference in ecosystems of the cheese surface versus those of the interior; the aerobic surface bacteria are generally G+C rich and represent diverse species, while the cheese interior comprises fewer species that are generally low in G+C content. TTGE and DGGE have proven here to be powerful methods to rapidly identify a broad range of bacterial species within dairy products.
机译：包括细菌，酵母和霉菌在内的许多微生物构成了牛奶和发酵乳制品中存在的复杂生态系统。我们的目的是描述各种奶酪的细菌生态系统，这些奶酪的生产技术不同，因此其细菌含量也不同。为此，我们开发了一种快速，半系统的方法，该方法基于具有暂时性温度梯度电泳（TTGE）的低G + C含量基因组细菌和变性梯度凝胶电泳（DGGE）的遗传谱分析（针对年龄较低和中等）的细菌G + C含量基因组。通过与约150种细菌的全面细菌参考数据库进行比较，为未知生态系统中的细菌分配了身份，其中包括有用的奶类微生物（乳酸菌），腐败菌（如假单胞菌和肠杆菌科）和病原菌（如单核细胞增多性李斯特菌）和金黄色葡萄球菌）。我们的分析提供了高分辨率的细菌，其包含不同商业奶酪的生态系统，并鉴定了常规方法无法识别的物种;在乳品生态系统中首次至少鉴定了两个属于嗜盐单胞菌和假单胞菌属的物种。我们的分析还揭示了奶酪表面与内部生态系统的惊人差异。有氧表面细菌通常富含G + C，代表各种物种，而奶酪内部包含的G + C含量通常较低的物种较少。事实证明，TTGE和DGGE是快速识别乳制品中各种细菌种类的有效方法。

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