scholarly journals Assessing the Occurrence of Waterborne Viruses in Reuse Systems: Analytical Limits and Needs

Pathogens ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 107 ◽  
Author(s):  
Charles P. Gerba ◽  
Walter Q. Betancourt
Keyword(s):  
Cell Culture ◽  
Virus Detection ◽  
Enteric Viruses ◽  
Enteric Virus ◽  
Culture Cell ◽  
Virus Infectivity ◽  
Recycled Water ◽  
Chemical Additives ◽  
Potable Reuse ◽  
Passage Number

Detection of waterborne enteric viruses is an essential tool in assessing the risk of waterborne transmission. Cell culture is considered a gold standard for detection of these viruses. However, it is important to recognize the uncertainty and limitations of enteric virus detection in cell culture. Cell culture cannot support replication of all virus types and strains, and numerous factors control the efficacy of specific virus detection assays, including chemical additives, cell culture passage number, and sequential passage of a sample in cell culture. These factors can result in a 2- to 100-fold underestimation of virus infectivity. Molecular methods reduce the time for detection of viruses and are useful for detection of those that do not produce cytopathogenic effects. The usefulness of polymerase chain reaction (PCR) to access virus infectivity has been demonstrated for only a limited number of enteric viruses and is limited by an understanding of the mechanism of virus inactivation. All of these issues are important to consider when assessing waterborne infectious viruses and expected goals on virus reductions needed for recycled water. The use of safety factors to account for this may be useful to ensure that the risks in drinking water and recycled water for potable reuse are minimized.

Application of Gene Probes to Virus Detection in Water

1989 ◽  
Vol 21 (3) ◽  
pp. 147-154 ◽  
Author(s):  
Charles P. Gerba ◽  
Aaron B. Margolin ◽  
Martinez J. Hewlett
Keyword(s):  
Cell Culture ◽  
Nucleic Acid ◽  
Virus Detection ◽  
Enteric Viruses ◽  
Serological Tests ◽  
Nucleic Acid Probes ◽  
Gene Probes ◽  
Detection Capabilities ◽  
Further Development ◽  
A Current

Gene probes offer a rapid and sensitive method for the detection of viruses in water and other environmental samples. Gene probes are small strands of nucleic acid labeled with radioactive or nonradioactive compounds for their detection. The target organism is identified by the hybridization of the probe to the organism's nucleic acid. Nucleic acid probes are at least 1000-fold more sensitive than serological tests such as enzyme-linked-immunoassay and do not first require cultivation of the virus for detection. Gene probes have been developed for organisms that do not grow in cell culture, and probes have been constructed for most of the major groups of enteric viruses. Gene probes have been applied to the detection of enteric viruses in water, marine sediment and shellfish. Radioactively labeled probes can detect as little as 1-10 infectious units of virus within 48 hours. A current disadvantage of probes is that they cannot determine the infectivity of the viruses; however, they can be used to quickly determine the growth of viruses in cell culture. Further development of nonradioactive probes should place virus detection capabilities into the hands of most water quality laboratories.

Enteric virus detection in Adriatic seawater by cell culture, polymerase chain reaction and polyacrylamide gel electrophoresis

1997 ◽  
Vol 31 (8) ◽  
pp. 1980-1984 ◽  
Author(s):  
Michele Muscillo ◽  
Annalaura Carducci ◽  
Giuseppina La Rosa ◽  
Laura Cantiani ◽  
Cinzia Marianelli
Keyword(s):  
Cell Culture ◽  
Polymerase Chain Reaction ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Virus Detection ◽  
Enteric Virus ◽  
Polyacrylamide Gel ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Comparative Study of Two Methods of Enteric Virus Detection and Enteric Virus Relationship with Bacterial Indicator in Poyang Lake, Jiangxi, China

2019 ◽  
Vol 16 (18) ◽  
pp. 3384 ◽  
Author(s):  
Wen ◽  
Zheng ◽  
Yuan ◽  
Zhu ◽  
Kuang ◽  
...  
Keyword(s):  
Water Quality ◽  
Poyang Lake ◽  
Virus Detection ◽  
Enteric Viruses ◽  
Enteric Virus ◽  
Jiangxi Province ◽  
Water Safety ◽  
Rt Pcr ◽  
Fecal Matter ◽  
Significant Difference

Currently, water contaminated with fecal matter poses a threat to public health and safety. Thus, enteric viruses are tested for as a part of water quality indicator assays; however, enteric viruses have not yet been listed in the criteria. Effective and sensitive methods for detecting enteric viruses are required in order to increase water safety. This study utilized enteric viruses as possible alternative indicators of water quality to examine fresh water in six sites in Poyang Lake, Nanchang, Jiangxi Province. The presence of norovirus geno-groups II (NoV GII), enteroviruses (EoV) and adenoviruses (AdV) were determined using Tianjin’s protocol and Hawaii’s protocol during a six month period from 2016–2017. The former used an electropositive material method for viral concentration and Taqman-q reverse transcription polymerase chain reaction (RT-PCR) to detect enteric viruses; while the latter used a filtration-based method for viral concentration and RT-PCR for enteric virus detection. There is a statistically significant difference between Tianjin’s method and Hawaii’s method for the detection of enteric viruses, such as NoV GII, EoV, and AdV (n = 36, p < 0.001). The enteric viruses showed no significant positive correlation with bacteria indicators (n = 36, p > 0.05). These data stress the need for additional indicators when establishing water quality systems, and the possibility of using enteric viruses as water quality indicators. It has become essential to improve shortcomings in order to search for an adequate method to detect enteric viruses in water and to implement such method in water quality monitoring.

Enteric viruses in drinking water supplies

2002 ◽  
Vol 2 (3) ◽  
pp. 17-22
Author(s):  
A.P. Wyn-Jones ◽  
J. Watkins ◽  
C. Francis ◽  
M. Laverick ◽  
J. Sellwood
Keyword(s):  
Drinking Water ◽  
Heavy Rainfall ◽  
Liquid Culture ◽  
Plaque Assay ◽  
Enteric Viruses ◽  
Enteric Virus ◽  
The Other ◽  
Raw Water ◽  
Rt Pcr ◽  
Water Supplies

Two rural spring drinking water supplies were studied for their enteric virus levels. In one, serving about 30 dwellings, the water was chlorinated before distribution; in the other, which served a dairy and six dwellings the water was not treated. Samples of treated (40 l) and untreated (20 l) water were taken under normal and heavy rainfall conditions over a six weeks period and concentrated by adsorption/elution and organic flocculation. Infectious enterovirus in concentrates was detected in liquid culture and enumerated by plaque assay, both in BGM cells, and concentrates were also analysed by RT-PCR. Viruses were found in both raw water supplies. Rural supplies need to be analysed for viruses as well as bacterial and protozoan pathogens if the full microbial hazard is to be determined.

Removal of PCR inhibiting substances in sewage sludge amended soil

1995 ◽  
Vol 31 (5-6) ◽  
pp. 311-315 ◽  
Author(s):  
Timothy M. Straub ◽  
Ian L. Pepper ◽  
Charles P. Gerba
Keyword(s):  
Cell Culture ◽  
Enteric Viruses ◽  
Treatment Method ◽  
Cytopathic Effects ◽  
Chelex 100 ◽  
Beef Extract ◽  
Polymerase Chain ◽  
Seminested Pcr ◽  
Soil Colloids ◽  
Amended Soil

Current methods for the detection of enteric viruses in soil involve elution of viruses from soil colloids using beef extract or other proteins. These eluates are then assayed in cell culture and observed daily for cytopathic effects (CPE). While this method is suitable for detection of enteric viruses by cell culture, these eluates contain humic acids and heavy metals that interfere with polymerase chain reaction (PCR) detection. Using beef extract eluates prepared from sludge amended soil, 10 different methods of eluate purification were evaluated for their ability to remove PCR inhibition and maximize sensitivity. The treatment method providing the greatest sensitivity of poliovirus detection by PCR was the combination of Sephadex G-50 and Chelex-100. Using this method 2 plaque forming units (PFU) could be detected after reverse transcription and 30 cycles of PCR. Thirty (30) cycles of seminested PCR were performed on these samples to verify nucleic acid sequences and increase sensitivity after the first 30 cycles of PCR. Using seminested PCR, sensitivity of detection using the Sephadex G-50 and Chelex-100 treatment method to 0.2 PFU. In addition to providing excellent sensitivity for viruses in sludge amended soils, this treatment method is relatively simple compared to other methods.

scholarly journals Evaluation of Murine Norovirus, Feline Calicivirus, Poliovirus, and MS2 as Surrogates for Human Norovirus in a Model of Viral Persistence in Surface Water and Groundwater

2007 ◽  
Vol 74 (2) ◽  
pp. 477-484 ◽  
Author(s):  
Jinhee Bae ◽  
Kellogg J. Schwab
Keyword(s):  
Cell Culture ◽  
Surface Water ◽  
Multiple Linear Regression Model ◽  
Viral Persistence ◽  
Environmental Stability ◽  
Great Promise ◽  
Feline Calicivirus ◽  
Virus Infectivity ◽  
Murine Norovirus ◽  
Surface Water And Groundwater

ABSTRACT Human noroviruses (NoVs) are a significant cause of nonbacterial gastroenteritis worldwide, with contaminated drinking water a potential transmission route. The absence of a cell culture infectivity model for NoV necessitates the use of molecular methods and/or viral surrogate models amenable to cell culture to predict NoV inactivation. The NoV surrogates murine NoV (MNV), feline calicivirus (FCV), poliovirus (PV), and male-specific coliphage MS2, in conjunction with Norwalk virus (NV), were spiked into surface water samples (n = 9) and groundwater samples (n = 6). Viral persistence was monitored at 25°C and 4°C by periodically analyzing virus infectivity (for all surrogate viruses) and nucleic acid (NA) for all tested viruses. FCV infectivity reduction rates were significantly higher than those of the other surrogate viruses. Infectivity reduction rates were significantly higher than NA reduction rates at 25°C (0.18 and 0.09 log10/day for FCV, 0.13 and 0.10 log10/day for PV, 0.12 and 0.06 log10/day for MS2, and 0.09 and 0.05 log10/day for MNV) but not significant at 4°C. According to a multiple linear regression model, the NV NA reduction rates (0.04 ± 0.01 log10/day) were not significantly different from the NA reduction rates of MS2 (0.05 ± 0.03 log10/day) and MNV (0.04 ± 0.03 log10/day) and were significantly different from those of FCV (0.08 ± 0.03 log10/day) and PV (0.09 ± 0.03 log10/day) at 25°C. In conclusion, MNV shows great promise as a human NoV surrogate due to its genetic similarity and environmental stability. FCV was much less stable and thus questionable as an adequate surrogate for human NoVs in surface water and groundwater.

scholarly journals Enteric Viruses and Inflammatory Bowel Disease

Viruses ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 104
Author(s):  
Georges Tarris ◽  
Alexis de Rougemont ◽  
Maëva Charkaoui ◽  
Christophe Michiels ◽  
Laurent Martin ◽  
...  
Keyword(s):  
Association Studies ◽  
Enteric Viruses ◽  
Enteric Virus ◽  
Viral Gastroenteritis ◽  
Blood Group Antigens ◽  
Genome Wide Association Studies ◽  
Virus Infections ◽  
T Helper ◽  
Inflammatory Bowel

Inflammatory bowel diseases (IBD), including ulcerative colitis (UC) and Crohn’s disease (CD), is a multifactorial disease in which dietary, genetic, immunological, and microbial factors are at play. The role of enteric viruses in IBD remains only partially explored. To date, epidemiological studies have not fully described the role of enteric viruses in inflammatory flare-ups, especially that of human noroviruses and rotaviruses, which are the main causative agents of viral gastroenteritis. Genome-wide association studies have demonstrated the association between IBD, polymorphisms of the FUT2 and FUT3 genes (which drive the synthesis of histo-blood group antigens), and ligands for norovirus and rotavirus in the intestine. The role of autophagy in defensin-deficient Paneth cells and the perturbations of cytokine secretion in T-helper 1 and T-helper 17 inflammatory pathways following enteric virus infections have been demonstrated as well. Enteric virus interactions with commensal bacteria could play a significant role in the modulation of enteric virus infections in IBD. Based on the currently incomplete knowledge of the complex phenomena underlying IBD pathogenesis, future studies using multi-sampling and data integration combined with new techniques such as human intestinal enteroids could help to decipher the role of enteric viruses in IBD.

scholarly journals Apparatus for Conditioning Unlimited Quantities of Finished Waters for Enteric Virus Detection

1974 ◽  
Vol 27 (6) ◽  
pp. 1177-1178 ◽  
Author(s):  
William F. Hill ◽  
Elmer W. Akin ◽  
William H. Benton ◽  
Charles J. Mayhew ◽  
Walter Jakubowski
Keyword(s):  
Virus Detection ◽  
Enteric Virus

scholarly journals Three-Year Study To Assess Human Enteric Viruses in Shellfish

2000 ◽  
Vol 66 (8) ◽  
pp. 3241-3248 ◽  
Author(s):  
F. Le Guyader ◽  
L. Haugarreau ◽  
L. Miossec ◽  
E. Dubois ◽  
M. Pommepuy
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Virus Detection ◽  
Enteric Viruses ◽  
Molecular Technique ◽  
Norwalk Virus ◽  
Viral Contamination ◽  
Long Period ◽  
Reverse Transcription Pcr ◽  
Human Enteric Viruses

ABSTRACT The main pathogenic enteric viruses able to persist in the environment, such as hepatitis A virus (HAV), Norwalk-like virus (NLV), enterovirus (EV), rotavirus (RV), and astrovirus (AV), were detected by reverse transcription-PCR and hybridization in shellfish during a 3-year study. Oyster samples (n = 108), occasionally containing bacteria, were less frequently contaminated, showing positivity for AV (17%), NLV (23%), EV (19%), and RV (27%), whereas mussel samples, collected in areas routinely impacted by human sewage, were more highly contaminated: AV (50%), HAV (13%), NLV (35%), EV (45%), and RV (52%). Sequences obtained from HAV and NLV amplicons showed a great variety of strains, especially for NLV (strains close to Mexico, Snow Mountain Agent, or Norwalk virus). Viral contamination was mainly observed during winter months, although there were some seasonal differences among the viruses. This first study of virus detection over a fairly long period of time suggests that routine analysis of shellfish by a molecular technique is feasible.

scholarly journals Diarrhea caused by rotavirus A, B, and C in suckling piglets from southern Brazil: molecular detection and histologic and immunohistochemical characterization

2018 ◽  
Vol 30 (3) ◽  
pp. 370-376 ◽  
Author(s):  
Paula R. Almeida ◽  
Elis Lorenzetti ◽  
Raquel S. Cruz ◽  
Tatiane T. Watanabe ◽  
Priscila Zlotowski ◽  
...  
Keyword(s):  
Virus Detection ◽  
Enteric Virus ◽  
Transmissible Gastroenteritis Virus ◽  
Southern Brazil ◽  
Rt Pcr ◽  
Viral Pathogen ◽  
Rotavirus A ◽  
Diarrhea Virus ◽  
Suckling Piglets ◽  
Transmissible Gastroenteritis

Rotavirus (RV) is an important viral pathogen causing diarrhea in piglets and other mammals worldwide. We describe 34 cases from 4 diarrheal outbreaks caused by RV in unvaccinated farrowing units in southern Brazil from 2011 to 2013. We performed autopsy, histologic examinations, bacterial culture, RV immunohistochemistry (IHC), and enteric virus detection through molecular assays for rotavirus A, B, and C, transmissible gastroenteritis virus, porcine epidemic diarrhea virus, sapovirus, norovirus, and kobuvirus. Histologically, villus atrophy (29 of 34) and epithelial vacuolation (27 of 34) occurred in all 4 outbreaks. Cell debris in the lamina propria occurred in 20 cases, mostly from outbreaks A (8 of 11), C (4 of 6), and D (7 of 11). IHC was positive for RV in 21 of 34 samples. RT-PCR was positive for RV in 20 of 30 samples; RV-C was the most frequently detected RV ( n = 17). Kobuvirus was detected in 11 samples, and, in 3 of them, there was single detection of this enteric virus.

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