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一种基于氧化还原染料 Resazurin 的下游工艺控制工具,用于快速准确地测量微生物代谢活性。

A downstream process control tool based on the redox dye resazurin for rapid and accurate measurement of microbial metabolic activity.

机构信息

Department of Civil Engineering, Yeungnam University, Gyeongsan, 38541, Republic of Korea.

出版信息

Anal Bioanal Chem. 2023 Sep;415(22):5539-5550. doi: 10.1007/s00216-023-04828-y. Epub 2023 Jul 4.

Abstract

Many sophisticated water treatment plants need a reliable, fast, and economical microbial load detection method. We refined a colorimetric assay using the redox dye resazurin to assess viable microorganisms. Here, we have used a mixed bacterial suspension of significant multi-drug-resistant coliform bacteria isolated from hospital wastewater and constructed a resazurin reduction calibration curve which could accurately predict the level of microbial contamination. The number of viable microorganisms was calculated from calibration curve in terms of log colony forming units (CFU) per milliliter. Ultrasonication disinfection of bacterial suspension for a duration of 50 min measured by resazurin assay depicted a reduction of 16.94%, 26.48%, and 37.69% at 410 W, 580 W, and 700 W, respectively. A synergistic effect of the combined methods of ultrasonication and heat disinfection treatments on raw wastewater and secondary wastewater effluent was observed and was also evaluated using both resazurin assay and standard plate count method. For raw wastewater, about 1.8 log reduction was observed for ultrasonication alone and 4 log CFU/mL reduction for thermosonication. In the secondary wastewater effluent, a reduction of 2.9 and 3.2 log CFU/mL was recorded for ultrasonication and thermosonication respectively. Resazurin microbial viability test results were highly comparable with conventional colony plate count for all treatment procedures, suggesting its appropriateness for quick and reliable wastewater sample microbial viability monitoring.

摘要

许多先进的水处理厂都需要一种可靠、快速且经济的微生物负荷检测方法。我们利用氧化还原染料-resazurin 改进了比色分析方法,以评估活微生物。在这里,我们使用了一种从医院废水中分离出的多药耐药大肠菌群的混合细菌悬浮液,构建了 resazurin 还原校准曲线,可以准确预测微生物污染水平。活微生物数量根据校准曲线以每毫升菌落形成单位(CFU)的对数计算。通过氧化还原染料法测量,细菌悬浮液经超声处理 50 分钟后,在 410 W、580 W 和 700 W 时,分别减少了 16.94%、26.48%和 37.69%。超声和热消毒处理的联合方法对原废水和二级废水处理均具有协同作用,并用氧化还原染料法和标准平板计数法进行了评估。对于原废水,单独超声处理可减少约 1.8 个对数,热声处理可减少 4 个对数 CFU/mL。在二级废水处理中,超声和热声处理分别记录到 2.9 和 3.2 个对数 CFU/mL 的减少。氧化还原染料微生物活力测试结果与所有处理程序的常规平板计数高度可比,表明其适用于快速可靠的废水样品微生物活力监测。

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