Application of Coagulation and Foam Concentration Method to Quantify Waterborne Pathogens in River Water Samples
One of the major challenges in detecting waterborne pathogens is the low concentration of the target bacteria in water. In this study, we applied the coagulation and foam concentration method to obtain DNA from water samples collected from upstream, near an estuary. The DNA samples were analyzed usi...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-11-01
|
| Series: | Water |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4441/14/22/3642 |
| _version_ | 1797463824142434304 |
|---|---|
| author | Yoshihiro Suzuki Atsushi Jikumaru Soichiro Tamai Kei Nukazawa Yoshifumi Masago Satoshi Ishii |
| author_facet | Yoshihiro Suzuki Atsushi Jikumaru Soichiro Tamai Kei Nukazawa Yoshifumi Masago Satoshi Ishii |
| author_sort | Yoshihiro Suzuki |
| collection | DOAJ |
| description | One of the major challenges in detecting waterborne pathogens is the low concentration of the target bacteria in water. In this study, we applied the coagulation and foam concentration method to obtain DNA from water samples collected from upstream, near an estuary. The DNA samples were analyzed using 16S rRNA gene sequencing to clarify the microbial community shifts and to identify potentially pathogenic bacteria. Bacterial communities changed as the river flowed downstream, most likely influenced by land use and human activities such as the discharge of wastewater-treatment plant effluent. Based on the 16S rRNA gene amplicon sequencing, potentially pathogenic bacteria were detected with greater than 0.1% of their relative abundances. Among these, <i>Yersinia ruckeri</i> and <i>Pseudomonas alcaligenes</i> were widely detected in the river water. In addition, digital PCR (dPCR) was used to quantify major waterborne pathogens. Shiga toxin-producing Escherichia coli (STEC), <i>Shigella</i> spp., and <i>Campylobacter jejuni</i> were all below the limit of detection. In contrast, general E. coli, which has the beta-D-glucuronidase gene (<i>uidA</i>) were detected by dPCR (copies/100 mL) at similar levels to those measured using the culture-based method (as colony forming units/100 mL). These results suggest that the coagulation and foam concentration method is useful for concentrating microbes and obtaining DNA from river water samples for environmental monitoring. |
| first_indexed | 2024-03-09T17:56:12Z |
| format | Article |
| id | doaj.art-662be082f5894548a0e64db594c252d3 |
| institution | Directory Open Access Journal |
| issn | 2073-4441 |
| language | English |
| last_indexed | 2024-03-09T17:56:12Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Water |
| spelling | doaj.art-662be082f5894548a0e64db594c252d32023-11-24T10:20:33ZengMDPI AGWater2073-44412022-11-011422364210.3390/w14223642Application of Coagulation and Foam Concentration Method to Quantify Waterborne Pathogens in River Water SamplesYoshihiro Suzuki0Atsushi Jikumaru1Soichiro Tamai2Kei Nukazawa3Yoshifumi Masago4Satoshi Ishii5Department of Civil and Environmental Engineering, Faculty of Engineering, University of Miyazaki, MiyazakI 889-2192, JapanDepartment of Civil and Environmental Engineering, Faculty of Engineering, University of Miyazaki, MiyazakI 889-2192, JapanDepartment of Civil and Environmental Engineering, Faculty of Engineering, University of Miyazaki, MiyazakI 889-2192, JapanDepartment of Civil and Environmental Engineering, Faculty of Engineering, University of Miyazaki, MiyazakI 889-2192, JapanCenter for Social and Environmental Systems Research, National Institute for Environmental Studies, Ibaraki 305-8506, JapanDepartment of Soil, Water, and Climate, University of Minnesota, Minneapolis, MN 55108-1095, USAOne of the major challenges in detecting waterborne pathogens is the low concentration of the target bacteria in water. In this study, we applied the coagulation and foam concentration method to obtain DNA from water samples collected from upstream, near an estuary. The DNA samples were analyzed using 16S rRNA gene sequencing to clarify the microbial community shifts and to identify potentially pathogenic bacteria. Bacterial communities changed as the river flowed downstream, most likely influenced by land use and human activities such as the discharge of wastewater-treatment plant effluent. Based on the 16S rRNA gene amplicon sequencing, potentially pathogenic bacteria were detected with greater than 0.1% of their relative abundances. Among these, <i>Yersinia ruckeri</i> and <i>Pseudomonas alcaligenes</i> were widely detected in the river water. In addition, digital PCR (dPCR) was used to quantify major waterborne pathogens. Shiga toxin-producing Escherichia coli (STEC), <i>Shigella</i> spp., and <i>Campylobacter jejuni</i> were all below the limit of detection. In contrast, general E. coli, which has the beta-D-glucuronidase gene (<i>uidA</i>) were detected by dPCR (copies/100 mL) at similar levels to those measured using the culture-based method (as colony forming units/100 mL). These results suggest that the coagulation and foam concentration method is useful for concentrating microbes and obtaining DNA from river water samples for environmental monitoring.https://www.mdpi.com/2073-4441/14/22/3642foam concentrationdigital PCRenvironmental DNApathogenic bacteriariver basin16S rRNA |
| spellingShingle | Yoshihiro Suzuki Atsushi Jikumaru Soichiro Tamai Kei Nukazawa Yoshifumi Masago Satoshi Ishii Application of Coagulation and Foam Concentration Method to Quantify Waterborne Pathogens in River Water Samples Water foam concentration digital PCR environmental DNA pathogenic bacteria river basin 16S rRNA |
| title | Application of Coagulation and Foam Concentration Method to Quantify Waterborne Pathogens in River Water Samples |
| title_full | Application of Coagulation and Foam Concentration Method to Quantify Waterborne Pathogens in River Water Samples |
| title_fullStr | Application of Coagulation and Foam Concentration Method to Quantify Waterborne Pathogens in River Water Samples |
| title_full_unstemmed | Application of Coagulation and Foam Concentration Method to Quantify Waterborne Pathogens in River Water Samples |
| title_short | Application of Coagulation and Foam Concentration Method to Quantify Waterborne Pathogens in River Water Samples |
| title_sort | application of coagulation and foam concentration method to quantify waterborne pathogens in river water samples |
| topic | foam concentration digital PCR environmental DNA pathogenic bacteria river basin 16S rRNA |
| url | https://www.mdpi.com/2073-4441/14/22/3642 |
| work_keys_str_mv | AT yoshihirosuzuki applicationofcoagulationandfoamconcentrationmethodtoquantifywaterbornepathogensinriverwatersamples AT atsushijikumaru applicationofcoagulationandfoamconcentrationmethodtoquantifywaterbornepathogensinriverwatersamples AT soichirotamai applicationofcoagulationandfoamconcentrationmethodtoquantifywaterbornepathogensinriverwatersamples AT keinukazawa applicationofcoagulationandfoamconcentrationmethodtoquantifywaterbornepathogensinriverwatersamples AT yoshifumimasago applicationofcoagulationandfoamconcentrationmethodtoquantifywaterbornepathogensinriverwatersamples AT satoshiishii applicationofcoagulationandfoamconcentrationmethodtoquantifywaterbornepathogensinriverwatersamples |