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

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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.

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Acknowledgements

This research was supported by the Basic Science Research Program through the National Research Foundation (NRF), funded by the Ministry of Education (2020R1I1A305481611), Republic of Korea. The authors thank the Research Center for Natural Products and Medical Materials (RCNM) for technical support regarding the nanoindentation test.

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

    Arkadeep Mukherjee & Young-Ho Ahn

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Mukherjee, A., Ahn, YH. A downstream process control tool based on the redox dye resazurin for rapid and accurate measurement of microbial metabolic activity. Anal Bioanal Chem 415, 5539–5550 (2023). https://doi.org/10.1007/s00216-023-04828-y

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