scholarly journals Quantitative RT-PCR Detection of Human Noroviruses and Hepatitis A Virus in Surface Water and Fresh Produce

2021 ◽  
Author(s):  
Mohamed N.F. Shaheen ◽  
Elmahdy M. Elmahdy ◽  
Lamiaa H. I. Mahmoud ◽  
Ibtisam A. Hammad ◽  
Elham R.S. Soliman
Keyword(s):  
Surface Water ◽  
Irrigation Water ◽  
Water Samples ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Fresh Produce ◽  
Surface Irrigation ◽  
Human Noroviruses ◽  
Human Enteric Viruses ◽  
Mean Concentration

Abstract Vegetables can be considered as a vehicle for the transmission of human enteric viruses such as noroviruses (genogroups I and II) and hepatitis A virus (HAV) when irrigated with contaminated irrigation or when prepared by infected food handlers. In the current study, we investigated the presence of hepatitis A virus (HAV) and human noroviruses (genogroups I and II) in fresh produce and surface water used in cultivation of this produce using real-time PCR. Samples were collected from six different points in the Mansoura and Giza regions, Egypt. Our analysis showed that at least one virus was found in 41.6% (30/72) of surface water samples and 27% (13/48) of fresh produce samples. HAV (23/72) with a mean viral concentration = 4 × 106 genome copies/litter (GC/L) was the most frequently identified virus in surface irrigation water samples, followed by human norovirus genogroup II (HNoV GII) (15/72, with a mean concentration = 1.2 × 106 GC/L, and human noroviruses genogroup I (HNoV GI) (12/72, with a mean concentration = 1.4 × 104 GC/L). Additionally, HAV (10/48) with a mean concentration = 5.2 × 105 genome copies/gram (GC/g) was also the most frequently detected virus in the fresh produce samples, followed by HNoV GII (8/48, with a mean concentration = 1.7 × 104 GC/g), meanwhile HNoV GI (6/48) was less detected virus with a mean concentration = 3 × 103 GC/g. This work suggests a wide prevalence of human enteric viruses in surface irrigation waters and fresh produce, which is of concern when the fresh produce is eaten raw. Thus, Additional monitoring for viral pathogens in irrigation water and food is needed to increase produce safely.

scholarly journals Molecular characterisation of hepatitis A virus strains from water sources in South Africa

2013 ◽  
Vol 69 (5) ◽  
pp. 923-933 ◽  
Author(s):  
R. Saïd ◽  
M. Wolfaardt ◽  
M. B. Taylor
Keyword(s):  
Surface Water ◽  
South African ◽  
Water Samples ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Water Sources ◽  
Nucleotide Sequence Analysis ◽  
Acid Extraction ◽  
Elution Method ◽  
Surface Water Samples

Hepatitis A virus (HAV) strains found in selected South African (SA) surface waters were characterised to establish what HAV types are circulating in the environment, thus reflecting circulation in the surrounding communities. Surface water samples used for irrigation or domestic purposes, and water samples from the outflow of wastewater plants were collected from six provinces. Viruses were recovered from the samples using a glass wool adsorption-elution method and then further concentrated using polyethylene glycol/sodium chloride precipitation. After automated nucleic acid extraction, samples were analysed for HAV by real-time reverse-transcriptase polymerase chain reaction. HAV strains were genotyped by nucleotide sequence analysis of the capsid gene VP1 and the VP1/P2B junction. HAVs were detected in 76% (16/21) of the surface water samples and in 37% (19/51) of the samples from the wastewater plants. Strains were characterised from 32 of the 35 samples and classified within genotype IB. The presence of genotype IB in the water sources confirms human faecal contamination. Hence, these faecally-contaminated water sources may be a potential transmission route of HAV infection and a potential source of contamination of irrigated fresh produce in SA.

Monitoring of human enteric viruses and coliform bacteria in waters after urban flood in Jakarta, Indonesia

2006 ◽  
Vol 54 (3) ◽  
pp. 203-210 ◽  
Author(s):  
C. Phanuwan ◽  
S. Takizawa ◽  
K. Oguma ◽  
H. Katayama ◽  
A. Yunika ◽  
...  
Keyword(s):  
Water Samples ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Tap Water ◽  
Enteric Viruses ◽  
Coliform Bacteria ◽  
High Concentration ◽  
Urban Flood ◽  
Human Enteric Viruses ◽  
Flooded Areas

Floodwaters in Kampung Melayu village, Jakarta, Indonesia, as well as river water and consumable water (including groundwater and tap water) samples in flooded and non-flooded areas, were quantitatively analysed to assess occurrence of viruses and total coliforms and E. coli as bacterial indicators after flooding event. High numbers of enterovirus, hepatitis A virus, norovirus (G1, G2) and adenovirus were detected at high concentration in floodwaters and waters sampled from Ciliwung River which runs across metropolitan Jakarta and is used widely for agriculture and domestic purposes by poor residents. One out of three groundwater wells in the flooded area was contaminated with all viruses tested while no viruses were found in groundwater samples in non-flooded areas and tap water samples. The results revealed that human enteric viruses, especially hepatitis A virus and adenovirus, were prevalent in Jakarta, Indonesia. This study suggested that flooding posed a higher risk of viral infection to the people through contamination of drinking water sources or direct contact with floodwaters.

Detection of Hepatitis a Virus and other Enteroviruses in Wastewater and Surface Water Samples by Gene Probe Assay

1991 ◽  
Vol 24 (2) ◽  
pp. 267-272 ◽  
Author(s):  
S. Dubrou ◽  
H. Kopecka ◽  
J. M. Lopez Pila ◽  
J. Maréchal ◽  
J. Prévot
Keyword(s):  
Surface Water ◽  
Cell Cultures ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Limit Of Detection ◽  
Blot Hybridization ◽  
Noncoding Region ◽  
Gene Probe ◽  
Dot Blot ◽  
Lower Efficiency

Enteroviruses were specifically detected by dot blot hybridization when using poliovirus type 1 (PV1) derived subgenomic radiolabeled cRNA probes (riboprobes) in environmental water specimens and in the cell cultures in which the viruses were amplificated. The riboprobe corresponding to the 5' noncoding sequence detected the majority of enteroviruses. Hepatitis A virus (HAV) was specifically detected by an HAV cRNA probe corresponding to the 5' noncoding region of its genome. By this test, the limit of detection of coxsackievirus B5 and echovirus 7 seeded in mineral water was 103 to 104 PFU/spot. In cell cultures, positive signals were observed in the lysates of cells infected by one PFU. Higher positive signals were obtained with a short PV1 probe (nt 221-670) corresponding to the 5' noncoding region, which is a well preserved sequence among the enteroviruses, than with PV1 genomic probe. Hybridization allowed a good detection of enteroviral RNAs in wastewater specimens, but with a lower efficiency in surface water. In this case, amplification of viruses in the cell cultures gave significant hybridization results.

Improved Detection of Norovirus and Hepatitis A Virus in Surface Water by Applying Pre-PCR Processing

2017 ◽  
Vol 9 (4) ◽  
pp. 395-405 ◽  
Author(s):  
Emmy Borgmästars ◽  
Mehrdad Mousavi Jazi ◽  
Sofia Persson ◽  
Linda Jansson ◽  
Peter Rådström ◽  
...  
Keyword(s):  
Surface Water ◽  
Hepatitis A ◽  
Hepatitis A 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 Recovery and Detection of Enteric Viruses from Non-Traditional Irrigation Water Sources

2019 ◽  
Vol 2 (3) ◽  
pp. 55 ◽  
Author(s):  
Brienna L. Anderson-Coughlin ◽  
Kalmia E. Kniel
Keyword(s):  
Irrigation Water ◽  
Water Samples ◽  
Hepatitis A Virus ◽  
Enteric Viruses ◽  
Environmental Water ◽  
Water Sources ◽  
Aichi Virus ◽  
Water Types ◽  
Virus Adsorption ◽  
Traditional Irrigation

The variability of environmental water samples impacts the allowance of one method to be universally ideal for all water types and volumes. Surface and reclaimed waters can be used for crop irrigation and may be referred to as non-traditional irrigation waters as these water types may be associated with a higher risk of microbial contamination compared to groundwater. These waters are typically more microbially and chemically complex than groundwater and have a higher risk of viral contamination. To detect viruses in these water types, an infinite number of variations can be made to traditional recovery methods. This protocol was developed based on a commonly used virus adsorption and elution (VIRADEL) method. Additional steps were included to simplify and efficiently reduce particulates in the viral concentrate and remove DNA from eluted nucleic acids prior to detection. Method alterations allow for volumes up to 40 liters to be processed with consistent recovery of enteric viruses including Aichi virus, hepatitis A virus, and noroviruses belonging to genogroups GI and GII. No inhibition was observed among either surface or reclaimed water samples. This protocol could be utilized in the monitoring of a wide array of irrigation water sources throughout irrigation processes.

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