Performance of viral and bacterial genetic markers for sewage pollution tracking in tropical Thailand

Watsawan Sangkaew; Akechai Kongprajug; Natcha Chyerochana; Warish Ahmed; Surapong Rattanakul; Thammanitchpol Denpetkul; Skorn Mongkolsuk; Kwanrawee Sirikanchana

doi:10.1016/j.watres.2020.116706

Performance of viral and bacterial genetic markers for sewage pollution tracking in tropical Thailand

Watsawan Sangkaew, Akechai Kongprajug, Natcha Chyerochana, Warish Ahmed, Surapong Rattanakul, Thammanitchpol Denpetkul, Skorn Mongkolsuk, Kwanrawee Sirikanchana^*

^*Corresponding author for this work

Randall Centre of Cell & Molecular Biophysics

Research output: Contribution to journal › Article › peer-review

22 Citations (Scopus)

Abstract

Identifying sewage contamination via microbial source tracking (MST) marker genes has proven useful for effective water quality management worldwide; however, performance evaluations for these marker genes in tropical areas are limited. Therefore, this research evaluated four human-associated MST marker genes (human polyomaviruses (JC and BK viruses [HPyVs]), bacteriophage crAssphage (CPQ_056), Lachnospiraceae Lachno3, and Bacteroides BacV6-21) for tracking sewage pollution in aquatic environments of Thailand. The viral marker genes, HPyV and crAssphage were highly sensitive and specific to sewage from onsite wastewater treatment plants (OWTPs; n = 19), with no cross-detection in 120 composite swine, cattle, chicken, duck, goat, sheep, and buffalo fecal samples. The bacterial marker genes, Lachno3 and BacV6-21, demonstrated high sensitivity but moderate specificity; however, using both markers could improve specificity to >0.80 (max value of 1.00). The most abundant markers in OWTP samples were Lachno3 and BacV6-21 (5.42–8.02 and nondetect–8.05 log₁₀ copies/100 mL), crAssphage (5.28–7.38 log₁₀ copies/100 mL), and HPyVs (3.66–6.53 log₁₀ copies/100 mL), respectively. Due to their increased specificity, the abundance of viral markers were further investigated in environmental waters, in which HPyVs showed greater levels (up to 4.33 log₁₀ copies/100 mL) and greater detection rates (92.7%) in two coastal beaches (n = 41) than crAssphage (up to 3.51 log₁₀ copies/100 mL and 56.1%). HPyVs were also found at slightly lower levels (up to 5.10 log₁₀ copies/100 mL), but at higher detection rates (92.6%), in a freshwater canal (n = 27) than crAssphage (up to 5.21 log₁₀ copies/100 mL and 88.9%). HPyVs and crAssphage marker genes were identified as highly sensitive and specific for tracking sewage pollution in aquatic environments of Thailand. This study underlines the importance of characterizing and validating MST markers in host groups and environmental waters before including them in a water quality management toolbox.

Original language	English
Article number	116706
Journal	WATER RESEARCH
Volume	190
DOIs	https://doi.org/10.1016/j.watres.2020.116706
Publication status	Published - 15 Feb 2021

Keywords

Fecal pollution
Freshwater
Indicator viruses
Microbial source tracking
Quantitative PCR
Seawater

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.watres.2020.116706

Cite this

@article{83ec4ca3aad640bfb7fa8027fece9c95,

title = "Performance of viral and bacterial genetic markers for sewage pollution tracking in tropical Thailand",

abstract = "Identifying sewage contamination via microbial source tracking (MST) marker genes has proven useful for effective water quality management worldwide; however, performance evaluations for these marker genes in tropical areas are limited. Therefore, this research evaluated four human-associated MST marker genes (human polyomaviruses (JC and BK viruses [HPyVs]), bacteriophage crAssphage (CPQ_056), Lachnospiraceae Lachno3, and Bacteroides BacV6-21) for tracking sewage pollution in aquatic environments of Thailand. The viral marker genes, HPyV and crAssphage were highly sensitive and specific to sewage from onsite wastewater treatment plants (OWTPs; n = 19), with no cross-detection in 120 composite swine, cattle, chicken, duck, goat, sheep, and buffalo fecal samples. The bacterial marker genes, Lachno3 and BacV6-21, demonstrated high sensitivity but moderate specificity; however, using both markers could improve specificity to >0.80 (max value of 1.00). The most abundant markers in OWTP samples were Lachno3 and BacV6-21 (5.42–8.02 and nondetect–8.05 log10 copies/100 mL), crAssphage (5.28–7.38 log10 copies/100 mL), and HPyVs (3.66–6.53 log10 copies/100 mL), respectively. Due to their increased specificity, the abundance of viral markers were further investigated in environmental waters, in which HPyVs showed greater levels (up to 4.33 log10 copies/100 mL) and greater detection rates (92.7%) in two coastal beaches (n = 41) than crAssphage (up to 3.51 log10 copies/100 mL and 56.1%). HPyVs were also found at slightly lower levels (up to 5.10 log10 copies/100 mL), but at higher detection rates (92.6%), in a freshwater canal (n = 27) than crAssphage (up to 5.21 log10 copies/100 mL and 88.9%). HPyVs and crAssphage marker genes were identified as highly sensitive and specific for tracking sewage pollution in aquatic environments of Thailand. This study underlines the importance of characterizing and validating MST markers in host groups and environmental waters before including them in a water quality management toolbox.",

keywords = "Fecal pollution, Freshwater, Indicator viruses, Microbial source tracking, Quantitative PCR, Seawater",

author = "Watsawan Sangkaew and Akechai Kongprajug and Natcha Chyerochana and Warish Ahmed and Surapong Rattanakul and Thammanitchpol Denpetkul and Skorn Mongkolsuk and Kwanrawee Sirikanchana",

note = "Funding Information: This work was financially supported by the Chulabhorn Research Institute , Thailand Science Research and Innovation (grant no. SRI6130703 ), and the Kurita Overseas Research Grant (grant no. 19Pth007 to A.K.) provided by Kurita Water and Environment Foundation. Publisher Copyright: {\textcopyright} 2020",

year = "2021",

month = feb,

day = "15",

doi = "10.1016/j.watres.2020.116706",

language = "English",

volume = "190",

journal = "WATER RESEARCH",

issn = "0043-1354",

publisher = "Elsevier Limited",

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TY - JOUR

T1 - Performance of viral and bacterial genetic markers for sewage pollution tracking in tropical Thailand

AU - Sangkaew, Watsawan

AU - Kongprajug, Akechai

AU - Chyerochana, Natcha

AU - Ahmed, Warish

AU - Rattanakul, Surapong

AU - Denpetkul, Thammanitchpol

AU - Mongkolsuk, Skorn

AU - Sirikanchana, Kwanrawee

N1 - Funding Information: This work was financially supported by the Chulabhorn Research Institute , Thailand Science Research and Innovation (grant no. SRI6130703 ), and the Kurita Overseas Research Grant (grant no. 19Pth007 to A.K.) provided by Kurita Water and Environment Foundation. Publisher Copyright: © 2020

PY - 2021/2/15

Y1 - 2021/2/15

N2 - Identifying sewage contamination via microbial source tracking (MST) marker genes has proven useful for effective water quality management worldwide; however, performance evaluations for these marker genes in tropical areas are limited. Therefore, this research evaluated four human-associated MST marker genes (human polyomaviruses (JC and BK viruses [HPyVs]), bacteriophage crAssphage (CPQ_056), Lachnospiraceae Lachno3, and Bacteroides BacV6-21) for tracking sewage pollution in aquatic environments of Thailand. The viral marker genes, HPyV and crAssphage were highly sensitive and specific to sewage from onsite wastewater treatment plants (OWTPs; n = 19), with no cross-detection in 120 composite swine, cattle, chicken, duck, goat, sheep, and buffalo fecal samples. The bacterial marker genes, Lachno3 and BacV6-21, demonstrated high sensitivity but moderate specificity; however, using both markers could improve specificity to >0.80 (max value of 1.00). The most abundant markers in OWTP samples were Lachno3 and BacV6-21 (5.42–8.02 and nondetect–8.05 log10 copies/100 mL), crAssphage (5.28–7.38 log10 copies/100 mL), and HPyVs (3.66–6.53 log10 copies/100 mL), respectively. Due to their increased specificity, the abundance of viral markers were further investigated in environmental waters, in which HPyVs showed greater levels (up to 4.33 log10 copies/100 mL) and greater detection rates (92.7%) in two coastal beaches (n = 41) than crAssphage (up to 3.51 log10 copies/100 mL and 56.1%). HPyVs were also found at slightly lower levels (up to 5.10 log10 copies/100 mL), but at higher detection rates (92.6%), in a freshwater canal (n = 27) than crAssphage (up to 5.21 log10 copies/100 mL and 88.9%). HPyVs and crAssphage marker genes were identified as highly sensitive and specific for tracking sewage pollution in aquatic environments of Thailand. This study underlines the importance of characterizing and validating MST markers in host groups and environmental waters before including them in a water quality management toolbox.

AB - Identifying sewage contamination via microbial source tracking (MST) marker genes has proven useful for effective water quality management worldwide; however, performance evaluations for these marker genes in tropical areas are limited. Therefore, this research evaluated four human-associated MST marker genes (human polyomaviruses (JC and BK viruses [HPyVs]), bacteriophage crAssphage (CPQ_056), Lachnospiraceae Lachno3, and Bacteroides BacV6-21) for tracking sewage pollution in aquatic environments of Thailand. The viral marker genes, HPyV and crAssphage were highly sensitive and specific to sewage from onsite wastewater treatment plants (OWTPs; n = 19), with no cross-detection in 120 composite swine, cattle, chicken, duck, goat, sheep, and buffalo fecal samples. The bacterial marker genes, Lachno3 and BacV6-21, demonstrated high sensitivity but moderate specificity; however, using both markers could improve specificity to >0.80 (max value of 1.00). The most abundant markers in OWTP samples were Lachno3 and BacV6-21 (5.42–8.02 and nondetect–8.05 log10 copies/100 mL), crAssphage (5.28–7.38 log10 copies/100 mL), and HPyVs (3.66–6.53 log10 copies/100 mL), respectively. Due to their increased specificity, the abundance of viral markers were further investigated in environmental waters, in which HPyVs showed greater levels (up to 4.33 log10 copies/100 mL) and greater detection rates (92.7%) in two coastal beaches (n = 41) than crAssphage (up to 3.51 log10 copies/100 mL and 56.1%). HPyVs were also found at slightly lower levels (up to 5.10 log10 copies/100 mL), but at higher detection rates (92.6%), in a freshwater canal (n = 27) than crAssphage (up to 5.21 log10 copies/100 mL and 88.9%). HPyVs and crAssphage marker genes were identified as highly sensitive and specific for tracking sewage pollution in aquatic environments of Thailand. This study underlines the importance of characterizing and validating MST markers in host groups and environmental waters before including them in a water quality management toolbox.

KW - Fecal pollution

KW - Freshwater

KW - Indicator viruses

KW - Microbial source tracking

KW - Quantitative PCR

KW - Seawater

UR - http://www.scopus.com/inward/record.url?scp=85097634435&partnerID=8YFLogxK

U2 - 10.1016/j.watres.2020.116706

DO - 10.1016/j.watres.2020.116706

M3 - Article

C2 - 33310444

AN - SCOPUS:85097634435

SN - 0043-1354

VL - 190

JO - WATER RESEARCH

JF - WATER RESEARCH

M1 - 116706

ER -

Performance of viral and bacterial genetic markers for sewage pollution tracking in tropical Thailand

Abstract

Keywords

UN SDGs

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