Assessing human-specific CrAssphage recovery after acidification-filtration concentrating method in environmental water

TY - JOUR

T1 - Assessing human-specific CrAssphage recovery after acidification-filtration concentrating method in environmental water

AU - Petcharat, Thitirat

AU - Kongprajug, Akechai

AU - Chyerochana, Natcha

AU - Sangkaew, Watsawan

AU - Mongkolsuk, Skorn

AU - Sirikanchana, Kwanrawee

N1 - Funding Information: This research was supported by the Chulabhorn Research Institute, Grant No. BT 2017-01. Publisher Copyright: © 2019 Water Environment Federation

PY - 2020/1/1

Y1 - 2020/1/1

N2 - Pinpointing water pollution sources using host-specific gastrointestinal microbes, known as microbial source tracking (MST), have significant benefits for countries with water quality management issues related to pollution. A recently discovered bacteriophage, crAssphage, shows promise as a human-specific MST marker. However, loss of genetic materials during the recovery and the detection processes could alter the ability to measure virus quantities in a water sample. This study determined the crAssphage recovery efficiencies in water sources, including seawater, freshwater, and influent and effluent from a wastewater treatment plant, by spiking natural crAssphage concentrates prior to DNA extraction and quantitative PCR analysis. The results showed that river and seawater with no or low crAssphage background experienced no recovery loss. Evaluating recovery efficiencies in samples with high crAssphage backgrounds posed a challenge due to the inability to prepare high crAssphage titers. This study highlights the importance of intra-laboratory assessment of recovery efficiency in environmental samples for retrieving absolute crAssphage quantification with correction of bias among water samples and increase in data accuracy. Practitioner points: In laboratory assessment of recovery efficiency is crucial for bias correction and data accuracy for absolute crAssphage quantification in water samples. No loss in crAssphage recovery was observed in river and seawater that contained no or low crAssphage backgrounds. Inability to prepare high crAssphage spike concentrations remains the major limitation for evaluating recovery in samples with high crAssphage backgrounds. The results underline the importance of evaluating method recovery in real environmental samples that reflect actual matrix effect. Absolute crAssphage quantification, as human-specific pollution marker, could be used for prioritizing water quality restoration and area-based management plan.

AB - Pinpointing water pollution sources using host-specific gastrointestinal microbes, known as microbial source tracking (MST), have significant benefits for countries with water quality management issues related to pollution. A recently discovered bacteriophage, crAssphage, shows promise as a human-specific MST marker. However, loss of genetic materials during the recovery and the detection processes could alter the ability to measure virus quantities in a water sample. This study determined the crAssphage recovery efficiencies in water sources, including seawater, freshwater, and influent and effluent from a wastewater treatment plant, by spiking natural crAssphage concentrates prior to DNA extraction and quantitative PCR analysis. The results showed that river and seawater with no or low crAssphage background experienced no recovery loss. Evaluating recovery efficiencies in samples with high crAssphage backgrounds posed a challenge due to the inability to prepare high crAssphage titers. This study highlights the importance of intra-laboratory assessment of recovery efficiency in environmental samples for retrieving absolute crAssphage quantification with correction of bias among water samples and increase in data accuracy. Practitioner points: In laboratory assessment of recovery efficiency is crucial for bias correction and data accuracy for absolute crAssphage quantification in water samples. No loss in crAssphage recovery was observed in river and seawater that contained no or low crAssphage backgrounds. Inability to prepare high crAssphage spike concentrations remains the major limitation for evaluating recovery in samples with high crAssphage backgrounds. The results underline the importance of evaluating method recovery in real environmental samples that reflect actual matrix effect. Absolute crAssphage quantification, as human-specific pollution marker, could be used for prioritizing water quality restoration and area-based management plan.

KW - crAssphage

KW - freshwater

KW - method recovery

KW - microbial source tracking

KW - sea water

KW - wastewater

UR - http://www.scopus.com/inward/record.url?scp=85071772497&partnerID=8YFLogxK

U2 - 10.1002/wer.1209

DO - 10.1002/wer.1209

M3 - Article

C2 - 31433097

AN - SCOPUS:85071772497

SN - 1061-4303

VL - 92

SP - 35

EP - 41

JO - WATER ENVIRONMENT RESEARCH

JF - WATER ENVIRONMENT RESEARCH

IS - 1

ER -