Flow cytometric quantification of viruses in activated sludge
Flow cytometric quantification of viruses in activated sludge
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DOI：
10.1016/j.watres.2014.10.018
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发表时间：
2015-01-01
期刊：
影响因子：
12.8
通讯作者：
Curtis, T. P.
中科院分区：
文献类型：
--
作者：
Brown, M. R.;Camezuli, S.;Curtis, T. P.
病毒可能在活性污泥系统的微生物动力学中发挥关键作用；然而，其量化的困难使得长期和大规模的研究成本高昂、及时且具有挑战性。因此，流式细胞术方案被优化并用于确定活性污泥样品中的病毒丰度。通过使用 Tween 80 和焦磷酸钠分离本土絮凝相关病毒，用 Tris-EDTA 稀释样品并用 SYBR Green II 染色，获得最佳流式细胞术特征和最高病毒计数。使用优化方案测定了 25 个活性污泥厂的病毒浓度，观察到的平均浓度为 2.35 x 10(9) mL(-1)。透射电子显微镜直接计数与流式细胞术计数高度相关 (p =
Viruses may play a critical role in the microbial dynamics of activated sludge systems; however the difficulty of their quantification makes long term and large scale studies costly, timely and challenging. Thus a flow cytometric protocol was optimised and employed to determine virus abundance in activated sludge samples. The best flow cytometry signature and highest virus count was obtained by separating the indigenous floc-associated viruses using Tween 80 and sodium pyrophosphate, diluting the sample with Tris-EDTA and staining with SYBR Green II. Using the optimised protocol viral concentrations from 25 activated sludge plants were determined, with average concentrations of 2.35 x 10(9) mL(-1) observed. Direct counts by transmission electron microscopy were highly correlated with flow cytometric counts (p =