Electronegative Filters Research Articles

Comparison of four concentration methods of adenovirus, norovirus and rotavirus in tap water

Human enteric viruses, as adenovirus (HAdV), norovirus (HuNoV) and rotavirus (RVA) are significant causes of gastroenteritis associated with consumption of contaminated water worldwide. Various methods have been described for their detection and monitoring in water. The aim of this study was to compare the performance of four conditions for concentrating HAdV, HuNoV and RVA from water matrices, in order to develop a single protocol that could simultaneously concentrate all target viruses from tap water. The tested conditions were based on the adsorption-elution using electronegative filters, in which we evaluated cation-coated filtration by MgCl2 with or without acid rinse by H2SO4 and two elution buffers, namely NaOH and tris-glycine-beef extract. Genomic material was extracted and amplified by real-time PCR and real-time RT-PCR using commercial kits. Based on the statistical analysis of amplification results (cycles of quantification), the condition involving cation-coated filtration by MgCl2 using electronegative filters with acid rinse by H2SO4 combined with NaOH elution allowed efficient recovery of both HAdV, HuNoV and RVA from tap water compared to the other conditions. These findings confirm the effectiveness of the approach used to monitor three major enteric viruses in tap water.

Food and Environmental Virology: Use of Passive Sampling to Characterize the Presence of SARS-CoV-2 and Other Viruses in Wastewater.

Fecal shedding of SARS-CoV-2 leads to a renaissance of wastewater-based epidemiology (WBE) as additional tool to follow epidemiological trends in the catchment of treatment plants. As alternative to the most commonly used composite samples in surveillance programs, passive sampling is increasingly studied. However, the many sorbent materials in different reports hamper the comparison of results and a standardization of the approach is necessary. Here, we compared different cost-effective sorption materials (cheesecloths, gauze swabs, electronegative filters, glass wool, and tampons) in torpedo-style housings with composite samples. Despite a remarkable variability of the concentration of SARS-CoV-2-specific gene copies, analysis of parallel-deposited passive samplers in the sewer demonstrated highest rate of positive samples and highest number of copies by using cheesecloths. Using this sorption material, monitoring of wastewater of three small catchments in the City of Dresden resulted in a rate of positive samples of 50% in comparison with composite samples (98%). During the investigation period, incidence of reported cases of SARS-CoV-2 in the catchments ranged between 16 and 170 per 100,000 persons and showed no correlation with the measured concentrations of E gene in wastewater. In contrast, constantly higher numbers of gene copies in passive vs. composite samples were found for human adenovirus and crAssphage indicating strong differences of efficacy of methods concerning the species investigated. Influenza virus A and B were sporadically detected allowing no comparison of results. The study contributes to the further understanding of possibilities and limits of passive sampling approaches in WBE.

Diurnal changes in pathogenic and indicator virus concentrations in wastewater

Wastewater-based epidemiology (WBE) has been commonly used for monitoring SARS-CoV-2 outbreaks. As sampling times and methods (i.e. grab vs composite) may vary, diurnal changes of viral concentrations in sewage should be better understood. In this study, we collected untreated wastewater samples hourly for 4 days at two wastewater treatment plants in Wales to establish diurnal patterns in virus concentrations and the physico-chemical properties of the water. Simultaneously, we also trialled three absorbent materials as passive samples as a simple and cost-efficient alternative for the collection of composite samples. Ninety-six percent of all liquid samples (n = 74) and 88% of the passive samplers (n = 59) were positive for SARS-CoV-2, whereas 87% and 97% of the liquid and passive samples were positive for the faecal indicator virus crAssphage, respectively. We found no significant daily variations in the concentration of the target viruses, ammonium and orthophosphate, and the pH and electrical conductivity levels were also stable. Weak positive correlations were found between some physico-chemical properties and viral concentrations. More variation was observed in samples taken from the influent stream as opposed to those taken from the influent tank. Of the absorbent materials trialled as passive samples, we found that tampons provided higher viral recoveries than electronegative filter paper and cotton gauze swabs. For all materials tested, viral recovery was dependent on the virus type. Our results indicate that grab samples may provide representative alternatives to 24-h composite samples if taken from the influent tank, hence reducing the costs of sampling for WBE programmes. Tampons are also viable alternatives for cost-efficient sampling; however, viral recovery should be optimised prior to use.

First quantitative detection of tomato brown rugose fruit virus in wastewater in Louisiana

We investigated the occurrence of tomato brown rugose fruit virus (ToBRFV) at a conventional wastewater treatment plant in Louisiana over a 13-month period, from March 2017 to March 2018. Influent, secondary effluent, and final effluent wastewater samples were collected monthly, and viruses were concentrated by the adsorption-elution method using an electronegative filter, followed by the detection using quantitative polymerase chain reaction. ToBRFV was detected in 10 (77 %) of 13 influent samples, 9 (69 %) of 13 in secondary effluent, and 6 (50 %) of 12 final effluents. The concentrations of ToBRFV in the influent samples ranged from 3.5 to 6.1 log10 copies/L and it was always higher than those in secondary or final effluents. Wastewater samples showed a high positive ratio of ToBRFV during fall and winter months. The findings highlight that routine monitoring of new viral indicator such as ToBRFV is necessary to understand its environmental distribution and correlation with pathogenic viruses. This is the first study providing quantitative data on the occurrence of ToBRFV in wastewater.

Detection of SARS-CoV-2 RNA in selected agricultural and food retail environments in Tehran, Iran.

The SARS-CoV-2 pandemic has and continues to impose a considerable public health burden. Although not likely foodborne, SARS-CoV-2 transmission has been well documented in agricultural and food retail environments in several countries, with transmission primarily thought to be worker-to-worker or through environmental high touch surfaces. However, the prevalence and degree to which SARS-CoV-2 contamination occurs in such settings in Iran has not been well documented. Furthermore, since SARS-CoV-2 has been observed to be shed in the feces of some infected individuals, wastewater has been utilized as a means of surveilling the occurrence of SARS-CoV-2 in some regions. This study aimed to investigate the presence of SARS-CoV-2 RNA along the food production and retail chain, from wastewater and irrigation water to vegetables in field and sold in retail. From September 2020 to January 2021, vegetables from different agricultural areas of Tehran province (n = 35), their irrigated agricultural water (n = 8), treated wastewater mixed into irrigated agricultural water (n = 8), and vegetables collected from markets in Tehran (n = 72) were tested for the presence of SARS-CoV-2 RNA. The vegetable samples were washed with TGBE buffer and concentrated with polyethylene glycol precipitation, while water samples were concentrated by an adsorption-elution method using an electronegative filter. RT-qPCR targeting the SARS-CoV-2 N and RdRp genes was then conducted. SARS-CoV-2 RNA was detected in 51/123 (41.5%) of the samples overall. The presence of SARS-CoV-2 RNA in treated wastewater, irrigation water, field vegetables, and market produce were 75, 37.5, 42.85, and 37.5%, respectively. These results indicate that SARS-CoV-2 RNA is present in food retail and may also suggest that produce can additionally be contaminated with SARS-CoV-2 RNA by agricultural water. This study demonstrates that SARS-CoV-2 RNA was detected in waste and irrigation water, as well as on produce both in field and at retail. However, more evidence is needed to understand if contaminated irrigation water causes SARS-CoV-2 RNA contamination of produce, and if there is a significant public health risk in consuming this produce.

Adsorption of SARS-CoV-2 onto granular activated carbon (GAC) in wastewater: Implications for improvements in passive sampling

Based on recent studies, passive sampling is a promising method for detecting SARS-CoV-2 in wastewater surveillance (WWS) applications. Passive sampling has many advantages over conventional sampling approaches. However, the potential benefits of passive sampling are also coupled with apparent limitations. We established a passive sampling technique for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in wastewater using electronegative filters. Though, it was evident that the adsorption capacity of the filters constrained their use. This work intends to demonstrate an optimized passive sampling technique for SARS-CoV-2 in wastewater using granular activated carbon (GAC). Through bench-scale batch-adsorption studies and sewershed deployments, we established the adsorption characteristics of SARS-CoV-2 and two human feacal viruses (PMMoV and CrAssphage) onto GAC. A pseudo-second-order model best-described adsorption kinetics for SARS-CoV-2 in either deionized (DI) water and SARS-CoV-2, CrAssphage, and PMMoV in wastewater. In both laboratory batch-adsorption experiments and in-situ sewershed deployments, the maximum amount of SARS-CoV-2 adsorbed by GAC occurred at ~60 h in wastewater. In wastewater, the maximum adsorption of PMMoV and CrAssphage by GAC occurred at ~60 h. In contrast, the adsorption capacity was reached in DI water seeded with SARS-CoV-2 after ~35 h. The equilibrium assay modeled the maximum adsorption quantity (qmax) in wastewater with spiked SARS-CoV-2 concentrations using a Hybrid Langmuir-Freundlich equation, a qmax of 2.5 × 109 GU/g was calculated. In paired sewershed deployments, it was found that GAC adsorbs SARS-CoV-2 in wastewater more effectively than electronegative filters. Based on the anticipated viral loading in wastewater, bi-weekly sampling intervals with deployments up to ~96 h are highly feasible without reaching adsorption capacity with GAC. GAC offers improved sensitivity and reproducibility to capture SARS-CoV-2 RNA in wastewater, promoting a scalable and convenient alternative for capturing viral pathogens in wastewater.

Comparison of Auto Sampling and Passive Sampling Methods for SARS-CoV-2 Detection in Wastewater.

Wastewater-based surveillance is emerging as an important tool for the COVID-19 pandemic trending. Current methods of wastewater collection, such as grab and auto-composite sampling, have drawbacks that impede effective surveillance, especially from small catchments with limited accessibility. Passive samplers, which are more cost-effective and require fewer resources to process, are promising candidates for monitoring wastewater for SARS-CoV-2. Here, we compared traditional auto sampling with passive sampling for SARS-CoV-2 detection in wastewater. A torpedo-style 3D-printed passive sampler device containing both cotton swabs and electronegative filter membranes was used. Between April and June 2021, fifteen passive samplers were placed at a local hospital’s wastewater outflow alongside an autosampler. Reverse transcription and quantitative polymerase chain reaction (RT-qPCR) was used to detect SARS-CoV-2 in the samples after processing and RNA extraction. The swab and membrane of the passive sampler showed similar detection rates and cycle threshold (Ct) values for SARS-CoV-2 RNA for the N1 and N2 gene targets. The passive method performed as well as the grab/auto sampling, with no significant differences between N1 and N2 Ct values. There were discrepant results on two days with negative grab/auto samples and positive passive samples, which might be related to the longer duration of passive sampling in the study. Overall, the passive sampler was rapid, reliable, and cost-effective, and could be used as an alternative sampling method for the detection of SARS-CoV-2 in wastewater.

Operational Constraints of Detecting SARS-CoV-2 on Passive Samplers using Electronegative Filters: A Kinetic and Equilibrium Analysis.

In developing an effective monitoring program for the wastewater surveillance of SARS-CoV-2 ribonucleic acid (RNA), the importance of sampling methodology is paramount. Passive sampling has been shown to be an effective tool to detect SARS-CoV-2 RNA in wastewater. However, the adsorption characteristics of SARS-CoV-2 RNA on passive sampling material are not well-understood, which further obscures the relationship between wastewater surveillance and community infection. In this work, adsorption kinetics and equilibrium characteristics were evaluated using batch-adsorption experiments for heat-inactivated SARS-CoV-2 (HI-SCV-2) adsorption to electronegative filters. Equilibrium isotherms were assessed or a range of total suspended solids (TSS) concentrations (118, 265, and 497 mg L–1) in wastewater, and a modeled qmax of 7 × 103 GU cm–2 was found. Surrogate adsorption kinetics followed a pseudo-first-order model in wastewater with maximum concentrations achieved within 24 h. In both field and isotherm experiments, equilibrium behavior and viral recovery were found to be associated with wastewater and eluate TSS. On the basis of the results of this study, we recommend a standard deployment duration of 24–48 h and the inclusion of eluate TSS measurement to assess the likelihood of solids inhibition during analysis.

Filtration, concentration and detection of salmonid alphavirus in seawater during a post-smolt salmon (Salmo salar) cohabitant challenge

Currently, the prevalence of salmonid alphavirus (SAV) in Norwegian Atlantic salmon farms is largely surveyed via sacrificing fish and sampling of organ tissue on a monthly basis. However, a more cost-efficient, straightforward, rapid, reliable, reproducible and animal welfare friendly method based on the detection of SAV in water could be considered as an alternative method. In the present study, such a method was developed and optimized through a 6 wk cohabitant challenge trial, using post-smolt Atlantic salmon Salmo salar L challenged with high or low doses of SAV subtype 3 (SAV3). Tank water and tissue samples from cohabitant fish were collected at 16 time points. SAV3 was concentrated from the water by filtration, using either electronegative or electropositive membrane filters, which were subsequently rinsed with one of 4 different buffer solutions. SAV3 was detected first in tank water (7 d post-challenge, DPC), and later in cohabitant fish organ tissue samples (12 DPC). The electronegative filter (MF-Millipore™) and rinsing with NucliSENS® easyMAG® Lysis Buffer presented the best SAV3 recovery. A significant positive correlation was found between SAV3 in the tank water concentrates and the mid-kidney samples. Based on these results, detection of SAV3 in filtrated seawater is believed to have the potential to serve as an alternative method for surveillance of SAV in Atlantic salmon farms.

Filtration, concentration and detection of salmonid alphavirus in seawater during a post-smolt salmon (Salmo salar) cohabitant challenge.

Currently, the prevalence of salmonid alphavirus (SAV) in Norwegian Atlantic salmon farms is largely surveyed via sacrificing fish and sampling of organ tissue on a monthly basis. However, a more cost-efficient, straightforward, rapid, reliable, reproducible and animal welfare friendly method based on the detection of SAV in water could be considered as an alternative method. In the present study, such a method was developed and optimized through a 6 wk cohabitant challenge trial, using post-smolt Atlantic salmon Salmo salar L challenged with high or low doses of SAV subtype 3 (SAV3). Tank water and tissue samples from cohabitant fish were collected at 16 time points. SAV3 was concentrated from the water by filtration, using either electronegative or electropositive membrane filters, which were subsequently rinsed with one of 4 different buffer solutions. SAV3 was detected first in tank water (7 d post-challenge, DPC), and later in cohabitant fish organ tissue samples (12 DPC). The electronegative filter (MF-Millipore™) and rinsing with NucliSENS® easyMAG® Lysis Buffer presented the best SAV3 recovery. A significant positive correlation was found between SAV3 in the tank water concentrates and the mid-kidney samples. Based on these results, detection of SAV3 in filtrated seawater is believed to have the potential to serve as an alternative method for surveillance of SAV in Atlantic salmon farms.

A novel passive sampling approach for SARS-CoV-2 in wastewater in a Canadian province with low prevalence of COVID-19

A simple passive sampling protocol using cheesecloth and electronegative filters coupled with a Tween®20-based elution technique provided reliable detection of SARS-CoV-2 in wastewater at targeted locations in a region of low COVID-19 prevalence.

Reduction of Cryptosporidium, Giardia, and Fecal Indicators by Bardenpho Wastewater Treatment.

Increased demand for water reuse and reclamation accentuates the importance for optimal wastewater treatment to limit protozoa in effluents. Two wastewater treatment plants utilizing advanced Bardenpho were investigated over a 12-month period to determine the incidence and reduction of Cryptosporidium, Giardia, Cyclospora, and fecal indicators. Results were compared to facilities that previously operated in the same geographical area. Protozoa (oo)cysts were concentrated using an electronegative filter and subsequently detected by fluorescent microscopy and/or PCR methods. Cryptosporidium and Giardia were frequently detected in raw sewage, but Cyclospora was not detected in any wastewater samples. Facilities with Bardenpho treatment exhibited higher removals of (oo)cysts than facilities utilizing activated sludge or trickling filters. This was likely due to Bardenpho systems having increased solid wasting rates; however, this mechanism cannot be confirmed as sludge samples were not analyzed. Use of dissolved-air-flotation instead of sedimentation tanks did not result in more efficient removal of (oo)cysts. Concentrations of protozoa were compared with each other, Escherichia coli, somatic coliphage, and viruses (pepper mild mottle virus, Aichi virus 1, adenovirus, and polyomaviruses JC and BK). Although significant correlations were rare, somatic coliphage showed the highest potential as an indicator for the abundance of protozoa in wastewaters.

Virus Reduction during Advanced Bardenpho and Conventional Wastewater Treatment Processes.

The present study investigated wastewater treatment for the removal of 11 different virus types (pepper mild mottle virus; Aichi virus; genogroup I, II, and IV noroviruses; enterovirus; sapovirus; group-A rotavirus; adenovirus; and JC and BK polyomaviruses) by two wastewater treatment facilities utilizing advanced Bardenpho technology and compared the results with conventional treatment processes. To our knowledge, this is the first study comparing full-scale treatment processes that all received sewage influent from the same region. The incidence of viruses in wastewater was assessed with respect to absolute abundance, occurrence, and reduction in monthly samples collected throughout a 12 month period in southern Arizona. Samples were concentrated via an electronegative filter method and quantified using TaqMan-based quantitative polymerase chain reaction (qPCR). Results suggest that Plant D, utilizing an advanced Bardenpho process as secondary treatment, effectively reduced pathogenic viruses better than facilities using conventional processes. However, the absence of cell-culture assays did not allow an accurate assessment of infective viruses. On the basis of these data, the Aichi virus is suggested as a conservative viral marker for adequate wastewater treatment, as it most often showed the best correlation coefficients to viral pathogens, was always detected at higher concentrations, and may overestimate the potential virus risk.

Relative abundance and treatment reduction of viruses during wastewater treatment processes — Identification of potential viral indicators

Waterborne pathogenic viruses discharged from wastewater treatment plants (WWTPs) pose potential public health risks. In the present study, we investigated the relative abundance, occurrence, and reduction of eleven different viruses at two WWTPs in southern Arizona over a 12-month period, from August 2011 to July 2012. Influent and effluent samples from the two WWTPs were collected monthly. Viruses were concentrated using an electronegative filter method and quantified using TaqMan-based quantitative PCR (qPCR) assays for each of the virus types (i.e., genogroup I, II and IV noroviruses, sapovirus, enterovirus, group A rotavirus, Aichi virus, pepper mild mottle virus, adenovirus, and JC and BK polyomaviruses), with murine norovirus internal control for the monitoring of extraction-RT-qPCR efficiencies. The pepper mild mottle virus, a plant virus, was found to be the most prevalent virus in both influent and effluent wastewater (annual mean concentration of 3.7–4.4×106copies/L and 4.6–6.3×105 copies/L in influent and effluent wastewater, respectively), showing a low reduction by the treatment processes (0.76–0.99 annual mean log10 reduction), and no significant seasonal change in concentration. Aichi virus, a human enteric virus, was also found in greater abundance, and showed lower reduction during wastewater treatment than other human enteric viruses. Our results suggest that these viruses could be used as potential indicators of wastewater reclamation system performance, with respect to virus occurrence and removal.

Occurrence of Cryptosporidium, Giardia, and Cyclospora in influent and effluent water at wastewater treatment plants in Arizona

We investigated the occurrence of Cryptosporidium, Giardia, and Cyclospora at two wastewater treatment plants (WWTPs) in Arizona over a 12-month period, from August 2011 to July 2012. Influent and effluent wastewater samples were collected monthly, and protozoan (oo)cysts were concentrated using an electronegative filter, followed by the detection of protozoa using fluorescent microscopy (Cryptosporidium oocysts and Giardia cysts) and PCR-based methods (Cryptosporidium spp., Giardia intestinalis, and Cyclospora cayetanensis). The concentration of Giardia cysts in the influent was always higher than that of Cryptosporidium oocysts (mean concentration of 4.8–6.4×103 versus 7.4×101–1.0×102(oo)cysts/l) with no clear seasonality, and log10 reduction of Giardia cysts was significantly higher than that of Cryptosporidium oocysts for both WWTPs (P<0.05). Log10 reduction of Giardia cysts at the WWTP utilizing activated sludge was significantly higher than the other WWTP using trickling filter (P=0.014), while no statistically significant difference between the two WWTPs was observed for the log10 reduction of Cryptosporidium oocysts (P=0.207). Phylogenetic analysis revealed that G. intestinalis strains identified in wastewater belonged to two assemblages, AII and B, which are potentially infectious to humans. C. cayetanensis was also detected from both influent and effluent using a newly developed quantitative PCR, with the highest influent concentration of 1.2×104copies/l. Our results demonstrated that these protozoan pathogens are prevalent in the study area and that efficacy of the conventional wastewater treatment processes at physically removing (oo)cysts is limited.

Detection of norovirus genogroup IV, klassevirus, and pepper mild mottle virus in sewage samples in South Korea

Norovirus (NoV) genogroup (G) IV has been infrequently isolated from patients suffering from acute gastroenteritis (AGE), although this virus has not been detected in Korea. Klassevirus, a novel virus belonging to the family Picornaviridae and a possible etiologic agent of AGE, and pepper mild mottle virus (PMMoV), which originates from processed pepper products and is shed in human feces, are suggested to be new indicators of fecal pollution. We aimed to investigate the presence of NoV-GIV, klassevirus, and PMMoV in sewage samples collected in Korea. Between December 2010 and February 2012, influent sewage samples were collected every month from a wastewater treatment plant located in the eastern part of Seoul in Korea. The sewage samples were concentrated by the adsorption elution method using an HA (pore size of 0.45μm with mixed cellulose ester) electronegative filter with an acid-rinse procedure. RT-PCR was performed using specific primers for the capsid gene of NoV-GII and NoV-GIV, the coat gene of PMMoV, and the VP0/VP1 gene of klassevirus. Among the 14 sewage samples tested, klassevirus was detected in eight (57.1%), PMMoV in eight (57.1%), NoV-GII in five (35.7%), and NoV-GIV in three (21.4%). NoV-GIV was detected in December 2010 and January and March 2011. PMMoV and klassevirus were frequently detected in winter. Phylogenetic analysis revealed that the NoV-GIV detected in this study belonged to G-IV1 lineage. This is the first study to confirm the presence of NoV-GIV, klassevirus, and PMMoV in sewage samples in Korea.

Methods for primary concentration of viruses from water samples: a review and meta-analysis of recent studies

Since the beginning of environmental virology in the mid-twentieth century, a key challenge to scientists in the environmental field has been how to collect, isolate and detect pathogenic viruses from water that is used for drinking and/or recreational purposes. Early studies investigated different types of membrane filters, with more sophisticated technologies being developed more recently. The purpose of this study was to look at the current state of the science of methods for the concentration of viruses from water. Several technologies were reviewed, and associated data were included in a meta-analysis which showed that electronegative filters, electropositive filters and ultrafilters are comparable in performance and that significant differences in recovery are due to virus type rather than filter type, water matrix or sample volume. This information is useful, as it will help to determine which method(s) should be used, particularly if there is a specific viral type being targeted for a particular study. In addition, it will be helpful when sampling different environmental water matrices and/or when budget allowance must be taken into consideration. Taken together, this will be useful in performing viral occurrence studies, which ultimately can help ensure safer water for both humans and the environment.

Concentration and Recovery of Viruses from Water: A Comprehensive Review

Enteric viruses are a cause of waterborne disease worldwide, and low numbers in drinking water can present a significant risk of infection. Because the numbers are often quite low, large volumes (100-1,000 L) of water are usually processed. The VIRADEL method using microporous filters is most commonly used today for this purpose. Negatively charged filters require the addition of multivalent salts and acidification of the water sample to effect virus adsorption, which can make large-volume sampling difficult. Positively charged filters require no preconditioning of samples, and are able to concentrate viruses from water over a greater pH range than electronegative filters. The most widely used electropositive filter is the Virosorb 1MDS; however, the Environmental Protection Agency has added the positively charged NanoCeram filters to their proposed Method 1615. Ultrafilters concentrate viruses based on size exclusion rather than electrokinetics, but are impractical for field sampling or processing of turbid water. Elution (recovery) of viruses from filters following concentration is performed with organic (e.g., beef extract) or inorganic solutions (e.g., sodium polyphosphates). Eluates are then reconcentrated to decrease the sample volume to enhance detection methods (e.g., cell culture infectivity assays and molecular detection techniques). While the majority of available filters have demonstrated high virus retention efficiencies, the methods to elute and reconcentrate viruses have met with varying degrees of success due to the biological variability of viruses present in water.

Molecular detection of three gastroenteritis viruses in urban surface waters in Beijing and correlation with levels of fecal indicator bacteria

To assess the presence of three gastroenteritis viruses responsible for human acute gastroenteritis in surface water, a 1-year study was carried out in the city of Beijing, China. A total of 108 urban surface water samples were collected from nine collection sites which were defined with a global positioning system in rivers or lakes from September 2006 to August 2007. The water samples were subjected to virus concentration using an HA electronegative filter, followed by polymerase chain reaction (PCR) for rotavirus (RV) astrovirus (AV), and norovirus (NV). It showed that the number of viruses detected in water samples from different sites was variable, totaling 63 virus strains, with rotavirus (48.1%) verified as the most prevalent detected, followed by astrovirus (AV, 5.6%), and norovirus (NV, 4.6%). RV was also quantified by real-time PCR and the concentration of RV ranged from 0 to 18.27 genome copies·L(-1). And the distributions of RV in surface water were abundant in cold weather (from September to February) while less prevailing in warm weather (from March to August). The high detection rate of RV we encountered in this study provided convincing evidence that RV circulated at a certain frequency in the Beijing population. There was no statistically significant correlation between RV levels and both fecal coliform (R (2) = 0.02) and Enterococcus faecalis (R (2) = 0.02) densities. Our study suggests prolonged virus persistence in aquatic environments and emphasizes the enteric virus group as the most reliable for environmental monitoring.

Detection of genogroup IV norovirus in wastewater and river water in Japan

To test wastewater and river water in Japan for genogroup IV norovirus (GIV NoV). Influent and effluent samples from a wastewater treatment plant and the Tamagawa River water samples were collected monthly for a year. The water samples were concentrated by the adsorption-elution method, using an HA electronegative filter with acid rinse procedure, followed by quantitative detection of GIV NoV using TaqMan-based real-time RT-PCR. Both wastewater and river water samples showed a high positive ratio of GIV NoV during winter and spring. The highest concentration in wastewater and river water was 6.9 x 10(4) and 1.5 x 10(4) copies l(-1), respectively. Presence of GIV NoV in the environments demonstrates that not only GI and GII NoVs but also GIV strains are circulating and that routine monitoring of GIV NoV in water environments is recommended to understand its epidemics, environmental distribution and potential health risks. This is the first study providing quantitative data on the occurrence of GIV NoV in environmental water over a 1-year period.