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Development of an Improved Selective Agar Medium for Isolation of Yersinia pestis

机译：分离鼠疫耶尔森氏菌的改良选择性琼脂培养基的开发

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Existing media designed for selective isolation of clinically important members of the genus Yersinia were found to be unsatisfactory for the growth and isolation of Yersinia pestis. We report the development of a new selective agar medium (termed BIN) that supports the growth of Y. pestis. The development of the formulation of this medium was based on a fluorescence screening system designed for monitoring bacterial growth on semisolid media, using a green fluorescent protein-expressing strain. High-throughput combinatorial experiments can be conducted for the quantitative evaluation of the effect of different medium components on growth. Generation of fluorescence plots in this system, using microplates, allowed the quantitative evaluation of the growth rate of Y. pestis EV76 cultures in different agar compositions. The final BIN formulation is based on brain heart infusion agar, to which the selective agents irgasan, cholate salts, crystal violet, and nystatin were introduced. It was found that BIN agar is more efficient in supporting colony formation and recovery of Y. pestis than are the conventional semisolid media MacConkey agar and Yersinia-selective agar (cefsulodin-irgasan-novobiocin agar). The advantage of BIN over other media has been also demonstrated in recovering virulent Y. pestis from the mixed bacterial populations found in decaying carcasses of infected mice. The BIN medium is suggested as a selective medium for isolation and recovery of Y. pestis from various backgrounds.
机译：发现设计用于选择性分离耶尔森氏菌属临床重要成员的现有培养基对鼠疫耶尔森氏菌的生长和分离不令人满意。我们报告了一种新的选择性琼脂培养基（称为BIN）的发展，该培养基可支持鼠疫耶尔森氏菌的生长。该培养基配方的开发基于荧光筛选系统，该系统设计为使用表达绿色荧光蛋白的菌株监测半固体培养基上的细菌生长。可以进行高通量组合实验，以定量评估不同培养基成分对生长的影响。使用微孔板在该系统中生成荧光图，可以定量评估不同琼脂组合物中鼠疫耶尔森氏菌EV76培养物的生长速率。最终的BIN配方基于脑心浸液琼脂，向其中引入了选择性试剂irgasan，胆酸盐，结晶紫和制霉菌素。发现BIN琼脂比常规半固体培养基MacConkey琼脂和耶尔森氏菌选择性琼脂（头孢磺隆-irgasan-novobiocin琼脂）更能有效地支持鼠疫耶尔森氏菌的菌落形成和恢复。与其他介质相比，BIN的优势还可以从感染的小鼠尸体的腐烂尸体中发现的混合细菌种群中回收有毒的鼠疫耶尔森氏菌。 BIN培养基被建议用作从各种背景分离和回收鼠疫耶尔森氏菌的选择性培养基。

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