Detection and distribution of vbnc/viable pathogenic bacteria in full-scale drinking water treatment plants

Guo, Lizheng; Wan, Kun; Zhu, Jianwen; Ye, Chengsong; Chabi, Kassim; Yu, Xin

doi:10.1016/j.jhazmat.2020.124335

Cited by 46 publications

(28 citation statements)

References 52 publications

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“…colony formation. Thus, molecular detection methods should be used to seek for pathogenic bacteria (Guo et al, 2021).…”

Section: Resuscitation Of Vbnc Cellsmentioning

confidence: 99%

“…Therefore, for detecting pathogenic bacteria, traditional culture‐dependent methods are not fully reliable for finding VBNC cells or those which do not develop visible growth, such as exemplified by micro‐colony formation. Thus, molecular detection methods should be used to seek for pathogenic bacteria (Guo et al ., 2021).…”

Section: Resuscitation Of Vbnc Cellsmentioning

confidence: 99%

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What do we mean by viability in terms of ‘viable but non‐culturable’ cells?

Chen

et al. 2021

Environ Microbiol Rep

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“…colony formation. Thus, molecular detection methods should be used to seek for pathogenic bacteria (Guo et al, 2021).…”

Section: Resuscitation Of Vbnc Cellsmentioning

confidence: 99%

Section: Resuscitation Of Vbnc Cellsmentioning

confidence: 99%

What do we mean by viability in terms of ‘viable but non‐culturable’ cells?

Chen

et al. 2021

Environ Microbiol Rep

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“…However, low DNA concentration or any genetic markers have been demonstrated to give high false-negative results, increased false discovery and artefactual results [44,52] that is likely to be the case for sequencing-based studies in chlorinated drinking water [29]. We found that many reported drinking water microbiome studies from chlorinated systems (including 16s rRNA amplicon sequencing/ metabarcoding, metagenomics) [20,[56][57][58][59][60][61][62][63][64] often exclude raw experimental data of extracted DNA concentrations that is crucial to assess the quality of the sequencing results.…”

Section: No Impact Of Water Volume On Dna Recoverymentioning

confidence: 87%

“…For example, comparison across samples with the same cell densities can correct taxa that appears to be a ‘rare’ genus [ 46 ]. Alternatively, qPCR (with and without propidium monoazide/PMA treatment) together with heterotrophic plate counts (HPC) can be applied to quantify total and viable opportunistic bacteria of concern in full scale chlorinated DWDSs [ 62 ]. We also foresee the increased importance of metaproteomics and metabolomics [ 94 , 95 ] to provide a complementary understanding of drinking water microbial communities.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of DNA extraction yield from a chlorinated drinking water distribution system

et al. 2021

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Desalination technology based on Reverse Osmosis (RO) membrane filtration has been resorted to provide high-quality drinking water. RO produced drinking water is characterized by a low bacterial cell concentration. Monitoring microbial quality and ensuring membrane-treated water safety has taken advantage of the rapid development of DNA-based techniques. However, the DNA extraction process from RO-based drinking water samples needs to be evaluated regarding the biomass amount (filtration volume) and residual disinfectant such as chlorine, as it can affect the DNA yield. We assessed the DNA recovery applied in drinking water microbiome studies as a function of (i) different filtration volumes, (ii) presence and absence of residual chlorine, and (iii) the addition of a known Escherichia coli concentration into the (sterile and non-sterile, chlorinated and dechlorinated) tap water prior filtration, and directly onto the (0.2 μm pore size, 47 mm diameter) mixed ester cellulose membrane filters without and after tap water filtration. Our findings demonstrated that the co-occurrence of residual chlorine and low biomass/cell density water samples (RO-treated water with a total cell concentration ranging between 2.47 × 102–1.5 × 103 cells/mL) failed to provide sufficient DNA quantity (below the threshold concentration required for sequencing-based procedures) irrespective of filtration volumes used (4, 20, 40, 60 L) and even after performing dechlorination. After exposure to tap water containing residual chlorine (0.2 mg/L), we observed a significant reduction of E. coli cell concentration and the degradation of its DNA (DNA yield was below detection limit) at a lower disinfectant level compared to what was previously reported, indicating that free-living bacteria and their DNA present in the drinking water are subject to the same conditions. The membrane spiking experiment confirmed no significant impact from any potential inhibitors (e.g. organic/inorganic components) present in the drinking water matrix on DNA extraction yield. We found that very low DNA content is likely to be the norm in chlorinated drinking water that gives hindsight to its limitation in providing robust results for any downstream molecular analyses for microbiome surveys. We advise that measurement of DNA yield is a necessary first step in chlorinated drinking water distribution systems (DWDSs) before conducting any downstream omics analyses such as amplicon sequencing to avoid inaccurate interpretations of results based on very low DNA content. This study expands a substantial source of bias in using DNA-based methods for low biomass samples typical in chlorinated DWDSs. Suggestions are provided for DNA-based research in drinking water with residual disinfectant.

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“…Today, vPCR technology is used in diverse research fields (e.g., in food, water, health, microbial ecology, agricultural soil, etc.) [25][26][27][28][29][30][31][32], it was shown that vPCR method can be used as a fast and effective viability test, as well as determining the microbial load in studies conducted in the field of environmental biotechnology and bioremediation. Observation of the viability by using the methods of molecular biology can be considered as the novelty and method applicability for the biodegradation studies.…”

Section: Discussionmentioning

confidence: 99%

Assessment of Comamonas testosteroni strain PT9 as a rapid phthalic acid degrader for industrial wastewaters

2021

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In this study, characterization of industry-borne Comamonas testosteroni strain PT9 isolate was performed by determining degradation ability on phthalic acid (PA). High-performance liquid chromatography analyses showed that strain PT9 completely degraded 102.94 mg/L of PA within 6 h. Viability polymerase chain reaction (vPCR) was performed with propidium monoazide treatment. vPCR showed that the PA has positively stimulated the cell growth during degradation. To consider the fate of PA, the proposed catalytic genes (ophA2, iphA2, tphA2, tphA3, pmdA, and pmdB) for the degradation pathways of PA isomers for C. testosteroni were screened in strain PT9. All genes except iphA2 were detected in strain PT9, and expression levels of related genes were analyzed by Real-Time PCR (qPCR).

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Detection and distribution of vbnc/viable pathogenic bacteria in full-scale drinking water treatment plants

Cited by 46 publications

References 52 publications

What do we mean by viability in terms of ‘viable but non‐culturable’ cells?

What do we mean by viability in terms of ‘viable but non‐culturable’ cells?

Evaluation of DNA extraction yield from a chlorinated drinking water distribution system

Assessment of Comamonas testosteroni strain PT9 as a rapid phthalic acid degrader for industrial wastewaters

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