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Experimental design for the optimization of propidium monoazide treatment to quantify viable and non-viable bacteria in piggery effluents

机译:优化单叠氮化丙啶处理以量化猪场废水中活菌和非活菌的实验设计

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摘要

BackgroundDistinguishing between viable and dead bacteria in animal and urban effluents is a major challenge. Among existing methods, propidium monoazide (PMA)-qPCR is a promising way to quantify viable cells. However, its efficiency depends on the composition of the effluent, particularly on total suspended solids (TSS)) and on methodological parameters. The aim of this study was evaluate the influence of three methodological factors (concentration of PMA, incubation time and photoactivation time) on the efficiency of PMA-qPCR to quantify viable and dead cells of Listeria monocytogenes used as a microorganism model, in two piggery effluents (manure and lagoon effluent containing 20 and 0.4 TSS g.kg−1, respectively). An experimental design strategy (Doehlert design and desirability function) was used to identify the experimental conditions to achieve optimal PMA-qPCR results.
机译:背景如何区分动物和城市污水中的活细菌和死细菌是一项重大挑战。在现有方法中,单叠氮化丙锭(PMA)-qPCR是定量活细胞的一种有前途的方法。但是,其效率取决于废水的成分,尤其取决于总悬浮固体(TSS)和方法学参数。这项研究的目的是评估三种方法学因素(PMA浓度,孵育时间和光激活时间)对两种猪场废水中PMA-qPCR定量用作微生物模型的单核细胞增生李斯特菌的活细胞和死细胞效率的影响。 (粪便和泻湖废水分别含20和0.4 TSS g.kg -1 )。实验设计策略(Doehlert设计和期望函数)用于确定实现最佳PMA-qPCR结果的实验​​条件。

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