Applicability of LIVE/DEAD BacLight stain with glutaraldehyde fixation for the measurement of bacterial abundance and viability in rainwater
Applicability of LIVE/DEAD BacLight stain with glutaraldehyde fixation for the measurement of bacterial abundance and viability in rainwater
复制标题
DOI：
10.1016/j.jes.2016.05.030
复制
发表时间：
2017-01-01
影响因子：
6.9
通讯作者：
Zhang, Daizhou
中科院分区：
文献类型：
--
作者：
Hu, Wei;Murata, Kotaro;Zhang, Daizhou
雨水含有大量细菌，雨水是细菌从大气传播到陆地和水面的有效途径。然而，由于缺乏可靠的方法，有关雨水细菌的定量信息非常有限。本研究采用荧光显微镜计数法和4‘，6-二氨基-2-苯基吲哚(DAPI)染色对雨水中活细菌和非活细菌的数量进行定量，以确定雨水中细菌的总数。结果表明，活/死BacLight染色的细菌总数与DAPI染色的结果一致，平均检测效率为(109+/-29)%。戊二醛固定组与未固定组细胞计数之比平均为(106+/-5)%。阴性对照的细菌浓度通常比雨水样本低一个数量级。但是，在降雨量较小的情况下，阴性对照的丰度可能比雨水样本的丰度高。这些结果表明，活/死BacLight细菌活力测定结合戊二醛固定和仔细的阴性对照调查是一种适用于雨水中细菌细胞浓度和活力测定的方法。(C)2016中国科学院生态环境科学研究中心。爱思唯尔出版公司(Elsevier B.V.)
Rainwater contains substantial bacteria and rain is an efficient pathway for the dissemination of bacteria from the atmosphere to land and water surfaces. However, quantitative information on rainwater bacteria is very limited due to the lack of a reliable method. In this study, the epifluorescence microscopy enumeration with the LIVE/DEAD BacLight Bacterial Viability Kit stain was verified to quantify the abundance of viable and non-viable bacterial cells in rainwater, with the 4',6-diamidino-2-phenylindole (DAPI) stain for the reference of total cell counts. Results showed that the total counts of bacterial cells by LIVE/DEAD BacLight staining were consistent with those by DAPI staining, and the average detection efficiency was (109 +/- 29)%. The ratio of cell count with glutaraldehyde fixation to that without fixation was (106 +/- 5)% on average. The bacterial concentration in negative control was usually an order of magnitude lower than that in rainwater samples. However, in case of small precipitation, the abundance in negative control could be more than that in rainwater samples. These results indicate that the enumeration with LIVE/DEAD BacLight bacterial viability assay coupled with glutaraldehyde fixation and careful negative control investigation is an approach applicable to the measurement of the concentration and viability of bacterial cells in rainwater. (C) 2016 The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V.