From Lab to Lake – Evaluation of Current Molecular Methods for the Detection of Infectious Enteric Viruses in Complex Water Matrices in an Urban Area

Leifels, Mats; Hamza, Ibrahim Ahmed; Krieger, Marion; Wilhelm, Michael; Mackowiak, Martin; Jurzik, Lars

doi:10.1371/journal.pone.0167105

Cited by 38 publications

(37 citation statements)

References 57 publications

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“…To retain diverse populations of crAssphage, each batch of stocks was prepared freshly by concentrating polluted canal water. Although different batches prepared could have contained different ratios of inactivated viruses to infectious viruses, viability detection of crAssphage as a water quality indicator is not as essential here as it is in cases of pathogenic viruses (Leifels et al, ).…”

Section: Resultsmentioning

confidence: 99%

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

Kongprajug

Chyerochana

Sangkaew

et al. 2019

Water Environment Research

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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.

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Section: Resultsmentioning

confidence: 99%

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

Kongprajug

Chyerochana

Sangkaew

et al. 2019

Water Environment Research

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“…However, this method has the limitation of not allowing the evaluation of virus viability and infectivity, defined as the capacity of the virus to infect host cells and to use their resources to produce new infectious virus particles. This aspect is important in assessing and understanding the risks to public health, because even if viral nucleic acids are detected by the PCR technique in water samples, they may not be able to infect human cells (191)(192)(193). Based on statistical correlations between genome copy numbers and infectious enteric viral particles in wastewater samples, Donia et al (194) proposed a cutoff value of 200 GC/liter to be used as an indication of EV survival in environmental monitoring (194).…”

Section: Enteric Virus Detection Methodsmentioning

confidence: 99%

Environmental and Adaptive Changes Necessitate a Paradigm Shift for Indicators of Fecal Contamination

et al. 2020

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Changes in the occurrence, distribution, and seasonal variation of waterborne pathogens due to global climate change may increase the risk of human exposure to these microorganisms, thus heightening the need for more reliable surveillance systems. Routine monitoring of drinking water supplies and recreational waters is performed using fecal indicator microorganisms, such as Escherichia coli, Enterococcus spp., and coliphages. However, the presence and numbers of these indicators, especially E. coli and Enterococcus spp., do not correlate well with those of other pathogens, especially enteric viruses, which are a major cause of waterborne outbreaks associated with contaminated water and food, and recreational use of lakes, ponds, rivers, and estuarine waters. For that reason, there is a growing need for a surveillance system that can detect and quantify viral pathogens directly in water sources to reduce transmission of pathogens associated with fecal transmission. In this review, we present an updated overview of relevant waterborne enteric viruses that we believe should be more commonly screened to better evaluate water quality and to determine the safety of water use and reuse and of epidemiological data on viral outbreaks. We also discuss current methodologies that are available to detect and quantify these viruses in water resources. Finally, we highlight challenges associated with virus monitoring. The information presented in this review is intended to aid in the assessment of human health risks due to contact with water sources, especially since current environmental and adaptive changes may be creating the need for a paradigm shift for indicators of fecal contamination. . Environmental and adaptive changes necessitate a paradigm shift for indicators of fecal contamination, Microbiol Spectrum 8(2):

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“…Em seguida realizou-se a extração e purificação do RNA com o QIAamp viral RNA Mini kit (Qiagen), seguindo as instruções do fabricante. RNA de Enterovirus já havia sido anteriormente detetado em massas de água para abastecimento e para fins recreativos de muitos outros países, nomeadamente, Estados Unidos, Japão, Holanda, Alemanha e África do Sul (Lodder et al 2010, Lee et al 2014, Lin & Sing 2015, Leifels et al 2016.…”

Section: Extração De Rna Para Deteção De Vírus Entéricos Por Rt-qpcrunclassified

Deteção de vírus entéricos em amostras de água natural superficial e de água para consumo humano

Salvador¹,

Caeiro²,

Aguilar³

et al. 2020

A&R

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Water-related infectious diseases are important causes of morbidity and mortality. Enterovirus and Hepatitis A and E viruses are, among others, representative of the enteric viruses, which are disease-causing agents mainly transmitted by the oral-fecal route, through water. The objective of this study is to search for enteric viruses by reverse transcription followed by Real-time PCR (RT-qPCR), in samples of natural surface water and in drinking water. It is also intended to evaluate the eventual association of these viruses with other fecal contamination indicators, and the efficacy of the water treatment plants in their elimination. It was confirmed the adequacy of the methodology implemented since, in the 15 samples analyzed so far, Hepatitis A virus and Enterovirus RNAs were detected in natural surface water samples, in two and three samples, respectively. Both viruses RNAs were detected in one of these samples. No viral nucleic acids were detected in drinking water samples. Fecal coliforms (microbial indicators of fecal contamination) were detected in natural water samples, but not in drinking water samples. Viral RNA and coliform detection only partially co-occurred.

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From Lab to Lake – Evaluation of Current Molecular Methods for the Detection of Infectious Enteric Viruses in Complex Water Matrices in an Urban Area

Cited by 38 publications

References 57 publications

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

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

Environmental and Adaptive Changes Necessitate a Paradigm Shift for Indicators of Fecal Contamination

Deteção de vírus entéricos em amostras de água natural superficial e de água para consumo humano

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