HUSCAP logo Hokkaido Univ. logo

Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Engineering / Faculty of Engineering >
Peer-reviewed Journal Articles, etc >

Simple and reliable enumeration of Escherichia coli concentrations in wastewater samples by measuring β-d-glucuronidase (GUS) activities via a microplate reader

This item is licensed under:Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International

Files in This Item:
STE136928.pdf767.17 kBPDFView/Open
Please use this identifier to cite or link to this item:http://hdl.handle.net/2115/83933

Title: Simple and reliable enumeration of Escherichia coli concentrations in wastewater samples by measuring β-d-glucuronidase (GUS) activities via a microplate reader
Authors: Satoh, Hisashi Browse this author →KAKEN DB
Kikuchi, Kai Browse this author
Katayose, Yutaka Browse this author
Tsuda, Shu Browse this author
Hirano, Reiko Browse this author
Hirakata, Yuga Browse this author
Kitajima, Masaaki Browse this author
Ishii, Satoshi Browse this author
Oshiki, Mamoru Browse this author →KAKEN DB
Hatamoto, Masashi Browse this author
Takahashi, Masahiro Browse this author
Okabe, Satoshi Browse this author →KAKEN DB
Keywords: Simple enumeration method
Fluorogenic enzyme substrate
Escherichia coli
Sewage wastewater
A microplate reader
Issue Date: 25-Jan-2020
Publisher: Elsevier
Journal Title: Science of The Total Environment
Volume: 715
Start Page: 136928
Publisher DOI: 10.1016/j.scitotenv.2020.136928
Abstract: Monitoring of Escherichia coli concentrations at wastewater treatment plants (WWTPs) is important to ensure process performance and protect public health. However, conventional E. coli enumeration methods are complicated and time- and labor-consuming. Here, we report a novel simple and reliable method based on β-D-glucuronidase (GUS) activity assay to enumerate E. coli concentrations in wastewater (WW) samples. An aliquot (20 μL) of the medium with fluorogenic enzyme substrate for E. coli and 180 μL of a WW sample were added to one well of a 96-well microplate. The microplate was placed in a microplate reader at 37°C. To this end, the fluorescence intensity of a fluorogenic enzyme substrate for E. coli was measured every 10 min over 3 h to determine GUS activity. The linear increase in the fluorescence intensity representing the GUS activities showed a positive correlation with E. coli concentrations in wastewater samples. However, the correlation equations were specific to WWTPs, which could be due to the difference in the E. coli population structures among WWTPs. We observed that the wastewater matrix is not a limitation to measure the GUS activity, and a WWTP-specific correlation equation can be used as a calibration curve to estimate the E. coli concentrations in the samples collected from that site. A comparison of the results with those of culture-dependent Colilert method proved that the current method is simple and useful for the enumeration of E. coli concentrations in wastewater samples reliably.
Rights: ©2020. This manuscript version is made available under the CC-BY-NC-ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/
https://creativecommons.org/licenses/by-nc-nd/4.0/
Type: article (author version)
URI: http://hdl.handle.net/2115/83933
Appears in Collections:工学院・工学研究院 (Graduate School of Engineering / Faculty of Engineering) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Export metadata:

OAI-PMH ( junii2 , jpcoar_1.0 )

MathJax is now OFF:


 

 - Hokkaido University