Comparative study of four extraction methods for enterovirus recovery from wastewater and sewage sludge

Belguith, Khaoula; Hassen, Abdennaceur; Aouni, Mahjoub

doi:10.1016/j.biortech.2005.03.022

Cited by 15 publications

(9 citation statements)

References 42 publications

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“…The result from this study (9.62 ϫ 10 2 copies per ml of raw wastewater) was similar to that of the Hewitt and colleagues study in New Zealand (34). The concentrations of HAdV 40/41 and EV in raw and secondary-treated wastewater were found to be similar to those previously reported (35)(36)(37).…”

Section: Discussionsupporting

confidence: 80%

Cross-Comparison of Human Wastewater-Associated Molecular Markers in Relation to Fecal Indicator Bacteria and Enteric Viruses in Recreational Beach Waters

Hughes

Beale

Dennis

et al. 2017

Appl Environ Microbiol

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Detection of human wastewater contamination in recreational waters is of critical importance to regulators due to the risks posed to public health. To identify such risks, human wastewater-associated microbial source tracking (MST) markers have been developed. At present, however, a greater understanding of the suitability of these markers for the detection of diluted human wastewater in environmental waters is necessary to predict risk. Here, we compared the process limit of detection (PLOD) and process limit of quantification (PLOQ) of six human wastewater-associated MST markers Among the six MST markers tested, HF183 was the most sensitive measure of human fecal pollution and was quantifiable up to dilutions of 10 Ϫ6 and 10 Ϫ4 for beach water samples seeded with raw and secondary-treated wastewater, respectively. Other markers and enteric viruses were detected at various dilutions (10 Ϫ1 to 10 Ϫ5 ). These MST markers, FIB, and enteric viruses were then quantified in beach water (n ϭ 12) and sand samples (n ϭ 12) from South East Queensland (SEQ), Australia, to estimate the levels of human fecal pollution. Of the 12 sites examined, beach water and sand samples from several sites had quantifiable concentrations of HF183 and PMMoV markers. Overall, our results indicate that while HF183 is the most sensitive measure of human fecal pollution, it should be used in conjunction with a conferring viral marker to avoid overestimating the risk of gastrointestinal illness.IMPORTANCE MST is an effective tool to help utilities and regulators improve recreational water quality around the globe. Human fecal pollution poses significant public health risks compared to animal fecal pollution. Several human wastewaterassociated markers have been developed and used for MST field studies. However, a head-to-head comparison in terms of their performance to detect diluted human fecal pollution in recreational water is lacking. In this study, we cross-compared the performance of six human wastewater-associated markers in relation to FIB and enteric viruses in beach water samples seeded with raw and secondary-treated wastewater. The results of this study will provide guidance to regulators and utilities on the appropriate application of MST markers for tracking the sources of human fecal pollution in environmental waters and confer human health risks.KEYWORDS microbial source tracking, human wastewater, fecal indicator bacteria, beach water, enteric viruses

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Section: Discussionsupporting

confidence: 80%

Cross-Comparison of Human Wastewater-Associated Molecular Markers in Relation to Fecal Indicator Bacteria and Enteric Viruses in Recreational Beach Waters

Hughes

Beale

Dennis

et al. 2017

Appl Environ Microbiol

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“…This non-significant variability in results from one site to another or on the same sampling site reflects an unmarked heterogeneity of the spatial and temporal distribution of viral particles. The heterogeneous distribution of EV was found in virtually all aquatic environments as revealed in several studies (Skraber et al 2004;Belguith et al 2006;Maul 2014). Discrepancy between positive rate in cell culture and positive rate in RT-PCR after culture suggested the presence of non-enterovirus CPE-causing viruses (such reovirus or adenovirus).…”

Section: Discussionmentioning

confidence: 97%

Effects of rainfalls variability and physical–chemical parameters on enteroviruses in sewage and lagoon in Yopougon, Côte d’Ivoire

et al. 2016

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The aim of this study was to assess the variability of the content of nutrients, oxidizable organic and particulate matters in raw sewage and the lagoon on the effect of rainfall. Then evaluate the impact of these changes in the concentration of enteroviruses (EVs) in waters. The sewage samples were collected at nine sampling points along the channel, which flows, into a tropical lagoon in Yopougon. Physical-chemical parameters (5-day Biochemical Oxygen Demand, Chemical Oxygen Demand, Suspended Particulate Matter, Total Phosphorus, Orthophosphate, Total Kjeldahl Nitrogen and Nitrate) as well as the concentration of EV in these waters were determined. The average numbers of EV isolated from the outlet of the channel were 9.06 9 10 4 PFU 100 ml -1 . Consequently, EV was present in 55.55 and 33.33 % of the samples in the 2 brackish lagoon collection sites. The effect of rainfall on viral load at the both sewage and brackish lagoon environments is significant correlate (two-way ANOVA, P \ 0.05). Furthermore, in lagoon environment, nutrients (Orthophosphate, Total Phosphorus), 5-day Biochemical Oxygen Demand, Chemical Oxygen Demand and Suspended Particulate Matter were significant correlated with EVs loads (P \ 0.05 by Pearson test). The overall results highlight the problem of sewage discharge into the lagoon and correlation between viral loads and water quality parameters in sewage and lagoon.

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“…The typing method was applied to 104 strains of HEV isolated from clinical samples sent to the Virology Unit of the University Hospital of Saint-Etienne (France) from 1977 to 2006 and to 12 strains of HEV recovered from environmental samples at the Faculty of Pharmacy of Monastir (Tunisia) from 1995 to 1997. The isolation technique was performed in both laboratories according to standard protocols as previously described (4,40). Once the enteroviral cytopathic effect involved more than 50% of the cell monolayer, the cells were scraped and an indirect immunofluorescence assay using the panenterovirus monoclonal antibody 5-D8/1 (DakoCytomation, Trappes, France) was performed to confirm the identification at the genus level (40).…”

Section: Methodsmentioning

confidence: 99%

Typing of Human Enterovirus by Partial Sequencing of VP2

Nasri

Bouslama

Omar³

et al. 2007

J Clin Microbiol

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The sequencing of the VP1 hypervariable region of the human enterovirus (HEV) genome has become the reference test for typing field isolates. This study describes a new strategy for typing HEV at the serotype level that uses a reverse transcription-PCR assay targeting the central part of the VP2 capsid protein. Two pairs of primers were used to amplify a fragment of 584 bp (with reference to the PV-1 sequence) or a part of it (368 bp) for typing. For a few strains not amplified by the first PCR, seminested primers enhanced the sensitivity (which was found to be approximately 10 ؊1 and 10 ؊4 50% tissue culture infective dose per reaction tube for the first and seminested assay, respectively). The typing method was then applied to 116 clinical and environmental strains of HEV. Sixty-one typeable isolates were correctly identified at the serotype level by comparison to seroneutralization. Forty-eight of 55 "untypeable" strains (87.3%) exhibited the same serotype using VP1 and VP2 sequencing methods. For six strains (four identified as EV-71, one as E-9, and one as E-30 by the VP2 method), no amplification was obtained by the VP1 method. The last strain, typed as CV-B4 by VP1 and CV-B3 by VP2 and monovalent antiserum, could exhibit recombination within the capsid region. Although the VP2 method was tested on only 36 of the 68 HEV serotypes, it appears to be a promising strategy for typing HEV strains isolated on a routine basis. The good sensitivity of the seminested technique could avoid cell culture and allow HEV typing directly from PCR products.Human enteroviruses (HEV) are among the most common of human viruses. Most infections are mild or asymptomatic, but some can lead to severe clinical presentations, especially in neonates and immunocompromised patients (32).The genus Enterovirus of the family Picornaviridae includes nonenveloped viruses comprising a 7,500-nucleotide singlestranded positive RNA genome protected by an icosahedral capsid. The genome encodes seven nonstructural proteins implicated in viral replication and maturation and four structural proteins, VP1 to VP4. VP1, VP2, and VP3 are located at the surface of the viral capsid and are exposed to immune pressure, whereas VP4 is located inside the capsid.The HEV serotypes were originally classified on the basis of antigenic properties and according to their natural and experimental pathogenesis: poliovirus (PV) infection in monkeys, coxsackievirus A (CV-A) and CV-B infection in suckling mice, and echovirus (E) infection in cell culture but not in mice (32). The molecular analysis of coding and noncoding regions (9) led to the classification of the 68 serotypes of HEV into five species (36): (i) HEV-A includes CV-A2, -3, -5 to -8, -10, -12, -14, and -16 and enterovirus 71 (EV-71) and -76; (ii) HEV-B includes CV-B1 to -6, CV-A9, and all Es, as well as EV-69, -73, -74, -75, -77, and -78; (iii) HEV-C includes CV-A1, -11, -13, -17, -20 to -22, and -24; (iv) EV-68 and -70 form the HEV-D group; and (v) the three serotypes of PV are still grouped into a separ...

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Comparative study of four extraction methods for enterovirus recovery from wastewater and sewage sludge

Cited by 15 publications

References 42 publications

Cross-Comparison of Human Wastewater-Associated Molecular Markers in Relation to Fecal Indicator Bacteria and Enteric Viruses in Recreational Beach Waters

Cross-Comparison of Human Wastewater-Associated Molecular Markers in Relation to Fecal Indicator Bacteria and Enteric Viruses in Recreational Beach Waters

Effects of rainfalls variability and physical–chemical parameters on enteroviruses in sewage and lagoon in Yopougon, Côte d’Ivoire

Typing of Human Enterovirus by Partial Sequencing of VP2

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