“…HF183/BacR287 values in the uncovered mesocosm were smaller than all 16 literature value inactivation rate constants whereas in the covered mesocosm, mean values were smaller than seven out of the nine literature value inactivation rate constants ( Fig. 4 ) (ranging from −0.03 to −2.55 d −1 ) ( Ahmed et al., 2014 , 2019 ; Bae and Wuertz, 2015 ; Balleste et al., 2018 , 2019 ; Dick et al., 2010 ; Eichmiller et al., 2014 ; Gilpin et al., 2013 ; Green et al., 2011 ; He et al., 2016 ; Jeanneau et al., 2012 ; Kirs et al., 2016 ; Liang et al., 2012 ; Walters and Field, 2009 ). Fig.…”
Fecal indicator bacteria (FIB) are typically used to monitor microbial water quality but are poor representatives of viruses due to different environmental fate. Viral fecal indicators have been proposed as alternatives to FIB; however, data evaluating the persistence of emerging viral fecal indicators under realistic environmental conditions is necessary to evaluate their potential application. In this study, we examined the persistence of five viral fecal indicators, including crAssphage and pepper mild mottle virus (PMMoV), and three bacterial fecal indicators (
E. coli
, enterococci and HF183/BacR287) in large-scale experimental ponds and freshwater mesocosms. Observed inactivation rate constants were highly variable and ranged from a minimum of −0.09 d
−1
for PMMoV to a maximum of −3.5 d
−1
for HF183/BacR287 in uncovered mesocosms. Overall, viral fecal indicators had slower inactivation than bacterial fecal indicators and PMMoV was inactivated more slowly than all other targets. These results demonstrate that bacterial fecal indicators inadequately represent viral fate following aging of sewage contaminated water due to differential persistence, and that currently used fecal indicator monitoring targets demonstrate highly variable persistence that should be considered during water quality monitoring and risk assessment.
“…HF183/BacR287 values in the uncovered mesocosm were smaller than all 16 literature value inactivation rate constants whereas in the covered mesocosm, mean values were smaller than seven out of the nine literature value inactivation rate constants ( Fig. 4 ) (ranging from −0.03 to −2.55 d −1 ) ( Ahmed et al., 2014 , 2019 ; Bae and Wuertz, 2015 ; Balleste et al., 2018 , 2019 ; Dick et al., 2010 ; Eichmiller et al., 2014 ; Gilpin et al., 2013 ; Green et al., 2011 ; He et al., 2016 ; Jeanneau et al., 2012 ; Kirs et al., 2016 ; Liang et al., 2012 ; Walters and Field, 2009 ). Fig.…”
Fecal indicator bacteria (FIB) are typically used to monitor microbial water quality but are poor representatives of viruses due to different environmental fate. Viral fecal indicators have been proposed as alternatives to FIB; however, data evaluating the persistence of emerging viral fecal indicators under realistic environmental conditions is necessary to evaluate their potential application. In this study, we examined the persistence of five viral fecal indicators, including crAssphage and pepper mild mottle virus (PMMoV), and three bacterial fecal indicators (
E. coli
, enterococci and HF183/BacR287) in large-scale experimental ponds and freshwater mesocosms. Observed inactivation rate constants were highly variable and ranged from a minimum of −0.09 d
−1
for PMMoV to a maximum of −3.5 d
−1
for HF183/BacR287 in uncovered mesocosms. Overall, viral fecal indicators had slower inactivation than bacterial fecal indicators and PMMoV was inactivated more slowly than all other targets. These results demonstrate that bacterial fecal indicators inadequately represent viral fate following aging of sewage contaminated water due to differential persistence, and that currently used fecal indicator monitoring targets demonstrate highly variable persistence that should be considered during water quality monitoring and risk assessment.
“…CrAssphage, a recently identified human fecal marker, was detected in all wastewater samples tested, at significantly higher concentrations than human enteric viruses and other indicator viruses. Previous studies also reported that crAssphage was highly abundant in various environmental samples 11,29,33,34,36,39 . Despite its abundance with high concentrations in water samples, several studies reported cross-reactions with feces from different animals 11,29,34,38 , raising questions about its suitability as a human fecal marker.…”
Section: Discussionmentioning
confidence: 76%
“…Moreover, the sequences of crAssphage variants detected in Europe were found to differ from those in the USA 11 . Both assays revealed the abundance of crAssphage of human origin; however, lower quantities (~3 log 10 lower) were also noted in samples of animal origin 11,29,33,36 , raising questions about the specificity of the assay. Although the appropriateness of crAssphage as a human fecal marker has been tested in many studies 11,29,[32][33][34][35]37,38 , to the best of our knowledge, no studies evaluating crAssphage as an indicator of virus reduction during water treatment processes have yet been reported.…”
this study was conducted to evaluate the applicability of crAssphage, pepper mild mottle virus (pMMoV), and tobacco mosaic virus (tMV) as indicators of the reduction of human enteric viruses during wastewater treatment. thirty-nine samples were collected from three steps at a wastewater treatment plant (raw sewage, secondary-treated sewage, and final effluent) monthly for a 13-month period. in addition to the three indicator viruses, eight human enteric viruses [human adenoviruses, Jc and BK polyomaviruses, Aichi virus 1 (AiV-1), enteroviruses, and noroviruses of genogroups I, II, and IV] were tested by quantitative pcR. indicator viruses were consistently detected in the tested samples, except for a few final effluents for crAssphage and TMV. The mean concentrations of crAssphage were significantly higher than those of most tested viruses. The concentrations of crAssphage in raw sewage were positively correlated with the concentrations of all tested human enteric viruses (p <0.05), suggesting the applicability of crAssphage as a suitable indicator to estimate the concentrations of human enteric viruses in raw sewage. The reduction ratios of AiV-1 (1.8 ± 0.7 log 10) were the lowest among the tested viruses, followed by TMV (2.0 ± 0.3 log 10) and PMMoV (2.0 ± 0.4 log 10). Our findings suggested that the use of not only AiV-1 and PMMoV but also TMV as indicators of reductions in viral levels can be applicable during wastewater treatment. Wastewater treatment plants (WWTPs) are currently facing numerous issues regarding insufficient treatment of human enteric viruses during the treatment process 1. Although a number of treatment procedures to remove physical, chemical, and microbiological waste from sewage are performed, these processess still need to be improved to reduce the viral content of water 2. The advanced multiple-barrier concept has been proposed for wastewater reclamation and reuse; however, it is insufficient to achieve complete reduction of human enteric viruses 3,4. In fact, advanced wastewater treatment has been developed to achieve reduction of viruses by facilitating chemical clarification, ultrafiltration, reverse osmosis, and advanced oxidation at different levels of regulatory control 5. However, despite implementing these modifications, the efficiency with which treatment can reduce viral levels in wastewater is still unsatisfactory. Moreover, no authorized regulatory standard for the reduction of viruses in wastewater treatment has yet been established 6. Human enteric viruses are the leading cause of waterborne illnesses and usually transmitted via the fecal-oral route 5-7. Infected individuals discharge millions of viral particles that ultimately enter sewage systems 7. In this context, it is important to study the efficiency of virus reduction at WWTPs to ascertain whether all viruses are removed from effluent samples or not. Fecal indicator bacteria, such as total coliforms and Escherichia coli, are not reliable indicators of the presence and removal of human enteric viruses during wastewa...
“…A higher abundance of bacterial markers than of viral markers was observed in sewage and in human feces (Ahmed et al, 2019a). Although faster decay rates of bacterial than of viral DNA markers have been reported in both freshwater and seawater (Ahmed et al, 2019c; Ballesté et al, 2019; Greaves et al, 2020), the higher abundance of bacterial markers in sewage could be due to the higher excretion rates of gut bacteria than of enteric viruses or to the better adsorption of viruses to particulate matters (Horswell et al, 2010).…”
Section: Discussionmentioning
confidence: 99%
“…CrAssphage (CPQ_056) was reported as up to six orders, expressed as copies/100 mL, in a secondary treated effluent in Louisiana, USA (Tandukar et al, 2020). Another crAssphage marker detected with a crAss-UP/LP assay, reported as four to six orders and expressed as copies/100 ml in WWTP-eff samples from Spain (Ballesté et al, 2019).…”
Identifying human sewage contamination via microbial source tracking (MST) marker genes has proven useful for effective water quality management worldwide; however, performance evaluations for these genes in tropical areas are limited. Therefore, this research assessed four human-associated MST marker genes in aquatic environments of Central Thailand: human polyomaviruses (JC and BK viruses [HPyVs]), bacteriophage crAssphage (CPQ_056), Lachnospiraceae Lachno3, and Bacteroides BacV6-21. HPyV and crAssphage assays were highly sensitive and specific to sewage (n = 19), with no cross-detection in 120 swine, cattle, chicken, duck, goat, sheep, and buffalo composite fecal samples. 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 sewage were Lachno3 and BacV6-21 (5.42-8.02 and non-detected-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. HPyVs showed higher levels (up to 4.33 log10 copies/100 mL) and higher 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%). Overall, both HPyVs and crAssphage are suggested as human sewage-associated MST markers in aquatic environments of Central 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.
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