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Ethidium and propidium monoazide: comparison of potential toxicity on Vibrio spp. viability
Letters in Applied Microbiology ( IF 2.0 ) Pub Date : 2020-11-15 , DOI: 10.1111/lam.13412
S. Copin 1 , J. Mougin 2 , V. Raguenet 1 , A. Robert‐Pillot 3 , G. Midelet 1 , T. Grard 2 , M. Bonnin‐Jusserand 2
Affiliation  

Vibrio spp., ubiquitous in the aquatic ecosystem, are bacteria of interest because of their involvement in human health, causing gastroenteritis after ingestion of seafood, as well as their role in vibriosis leading to severe losses in aquaculture production. Their ability to enter a viable but non-culturable (VBNC) state under stressful environmental conditions may lead to underestimation of the Vibrio population by traditional microbiological enumeration methods. As a result, using molecular methods in combination with EMA or PMA allows the detection of viable (VBNC and culturable viable) cells. In this study, the impact of the EMA and PMA was tested at different concentrations on the viability of several Vibrio species. We compared the toxicity of these two DNA-binding dyes to determine the best pretreatment to use with qPCR to discriminate between viable and dead Vibrio cells. Our results showed that EMA displayed lethal effects for each strain of V. cholerae and V. vulnificus tested. In contrast, the concentrations of PMA tested had no toxic effect on the viability of Vibrio cells studied. These results may help to achieve optimal PMA-qPCR methods to detect viable Vibrio spp. cells in food and environmental samples.

中文翻译:

乙锭和丙锭单叠氮化物:对弧菌属潜在毒性的比较。可行性

弧菌属普遍存在于水生生态系统中,是一种令人感兴趣的细菌,因为它们参与人类健康,摄入海鲜后会引起胃肠炎,以及它们在弧菌病中的作用导致水产养殖生产的严重损失。它们在压力环境条件下进入可行但不可培养 (VBNC) 状态的能力可能会导致传统微生物计数方法低估弧菌种群。因此,将分子方法与 EMA 或 PMA 结合使用可以检测活细胞(VBNC 和可培养活细胞)。在本研究中,测试了不同浓度的 EMA 和 PMA 对几种弧菌物种生存力的影响。我们比较了这两种 DNA 结合染料的毒性,以确定与 qPCR 一起使用的最佳预处理,以区分活的和死的弧菌细胞。我们的结果表明,EMA 对每个测试的霍乱弧菌和创伤弧菌菌株都显示出致死作用。相比之下,所测试的 PMA 浓度对所研究的弧菌细胞的活力没有毒性影响。这些结果可能有助于实现最佳 PMA-qPCR 方法来检测可行的弧菌属。食品和环境样品中的细胞。
更新日期:2020-11-15
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