Enumeration of water-borne bacteria using viability assays and flow cytometry: a comparison to culture-based techniques
Enumeration of water-borne bacteria using viability assays and flow cytometry: a comparison to culture-based techniques
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DOI：
10.1016/s0167-7012(03)00201-x
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发表时间：
2003-12-01
影响因子：
2.2
通讯作者：
Saint, CP
中科院分区：
文献类型：
--
作者：
Hoefel, D;Grooby, WL;Saint, CP
保持集水区或配水系统的最佳条件在很大程度上依赖于水务部门能否获得快速、准确的水质数据，包括细菌质量的指示。在这项研究中，BacLight(TM) 细菌活力试剂盒和羧基荧光素二乙酸酯 (CFDA) 与流式细胞术 (FCM) 结合使用，可快速检测南澳大利亚各地原水和饮用水中的生理活性细菌。除了基于培养的异养平板计数 (HPC) 方法和商业 SimPlate(TM) 技术之外，还将结果与直接活菌计数 (DVC) 和定量 DVC (qDVC) 进行比较。原水和饮用水分析表明，与使用快速 FCM 检测检测到的生理活性细菌数量相比，DVC 和基于培养的技术报告的活细菌数量显着减少，其中这种差异在不同样品中似乎是非线性的。使用 qDVC 作为活力测定获得了不确定的结果。特别是，HPC 结果比原水 FCM 检测报告的结果低 2-4 个对数级。尽管FCM 检测报告的活性细菌数量在5.56 x 10(2) 至3.94 x 10(4) ml(-1) 之间，但经HPC 检测，饮用水中几乎没有细菌可培养。这些差异可能归因于非异养细菌的存在、亚致死损伤或采用活跃但不可培养 (ABNC) 状态。 (C) 2003 Elsevier B.V 保留所有权利。
Maintaining optimal conditions in catchments or distribution systems relies heavily on water authorities having access to rapid and accurate water quality data, including an indication of bacteriological quality. In this study, the BacLight(TM) bacterial viability kit and carboxyfluorescein diacetate (CFDA) were coupled with flow cytometry (FCM) for rapid detection of physiologically active bacteria from raw and potable waters taken from various locations around South Australia. Results were compared to the direct viable count (DVC) and quantitative DVC (qDVC), in addition to the culture-based methods of the heterotrophic plate count (HPC) and a commercial SimPlate(TM) technique. Raw and potable water analysis revealed that DVC and culture-based techniques reported significantly fewer viable bacteria compared to the number of physiologically active bacteria detected using the rapid FCM assays, where this difference appeared to be nonlinear across different samples. Inconclusive results were obtained using qDVC as a viability assay. In particular, HPC results were 2-4 log orders of magnitude below that reported by the FCM assays for raw waters. Few bacteria in potable waters examined were culturable by HPC, even though FCM assays reported between 5.56 x 10(2) and 3.94 x 10(4) active bacteria ml(-1). These differences may be attributed to the presence of nonheterotrophic bacteria, sublethal injury or the adoption of an active but nonculturable (ABNC) state. (C) 2003 Elsevier B.V All rights reserved.