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)
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