Sawaya K, Kaneko N, Fukushi K, Yaguchi J
Advanced Course of Civil and Environmental Engineering, Hachinohe National College of Technology, 16-1 Uwanotai, Tamonoki, Hachinohe 039-1192, Japan.
Water Sci Technol. 2008;58(7):1343-8. doi: 10.2166/wst.2008.445.
Direct microscopic methods using several fluorescent staining were applied to estimate the proportion of physiologically active bacteria in the water environment and evaluate the efficacy of disinfection with chlorine. 4',6-diamidino-2-phenylindole (DAPI) was used to determine total bacterial numbers, and 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) was chosen for direct detection of respiring bacteria. BacLight kit was used to assess bacterial membrane integrity. Bacteria with growth potential were enumerated using the DVC method and microcolony technique. The total bacterial number in river was 8 x 10(6)-3 x 10(10) cells/mL, and colony forming units on R2A medium were 1 x 10(4)-4 x 10(5) cfu/mL. In the case of wastewater treatment plant, 1-10% of total bacterial cells could form colonies. Physiologically active bacteria in river and wastewater treatment plant determined by fluorescent staining were much higher than those obtained by plate counting. The effect of chlorine on the physiological viability of Escherichia coli was also investigated. Microscopic viable bacteria were even more chlorine resistant than culturable bacteria. The inactivation rate coefficients of direct viable bacteria were one-second to third those of culturable bacteria.
采用几种荧光染色的直接显微镜方法来估计水环境中生理活性细菌的比例,并评估氯消毒的效果。使用4',6-二脒基-2-苯基吲哚(DAPI)来确定细菌总数,选择5-氰基-2,3-二甲基氯化四氮唑(CTC)直接检测呼吸细菌。使用BacLight试剂盒评估细菌膜完整性。使用DVC方法和微菌落技术对具有生长潜力的细菌进行计数。河流中的细菌总数为8×10⁶ - 3×10¹⁰个细胞/毫升,R2A培养基上的菌落形成单位为1×10⁴ - 4×10⁵ cfu/毫升。对于污水处理厂,1 - 10%的细菌细胞能够形成菌落。通过荧光染色确定的河流和污水处理厂中的生理活性细菌远高于平板计数法获得的结果。还研究了氯对大肠杆菌生理活力的影响。显微镜下的活菌比可培养细菌对氯的耐受性更强。直接活菌的灭活速率系数是可培养细菌的三分之一到二分之一。