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Enteric Virus Detection: Recent Developments and Application in Food and Waters

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A special issue of Microorganisms (ISSN 2076-2607). This special issue belongs to the section "Food Microbiology".

Deadline for manuscript submissions: closed (31 July 2020) | Viewed by 16483

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Dr. Gloria Sanchez

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Environmental Virology and Food Safety Lab (VISAFELab), Department of Preservation and Food Safety Technologies, IATA-CSIC, Av. Agustín Escardino 7, 46980 Valencia, Spain
Interests: emerging viruses; molecular techniques; food safety
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Enteric virus contamination, particularly norovirus and hepatitis A and E viruses, in food and water is a global public health concern. Because of the increasing number of viral food- and waterborne outbreaks, it has become even more important to have reliable, standardized, and widely applicable techniques for their detection, quantification, and characterization in food and water. Since their infectious dose is very low, sensitive methods are therefore needed when screening food products and water samples for viruses. In such a scenario, improvements to the currently available procedures must be undertaken in order to assess the occurrence of the most relevant viruses in food and water matrices.

The current Special Issue will publish state-of-the-art research and review papers on recent development and application of molecular technologies used for monitoring human enteric viruses in food and water samples. We specifically welcome manuscripts discussing recent developments on methods for improving virus concentration and molecular techniques, such as digital PCR and next-generation sequencing, in different types of food matrices and environmental waters. Furthermore, manuscripts investigating the occurrence of the most relevant human enteric viruses in food products and water are also welcome in this issue.

Dr. Gloria Sanchez
Guest Editor

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Keywords

enteric viruses
norovirus
hepatitis A virus
hepatitis E virus
molecular detection methods
foodborne
waterborne
occurrence

Published Papers (5 papers)

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Research

12 pages, 1833 KiB

Open AccessArticle

Interlaboratory Validation of a Detection Method for Hepatitis E Virus RNA in Pig Liver

by Eva Trojnar, Matthias Contzen, Dominik Moor, Anja Carl, Sabine Burkhardt, Jochen Kilwinski, Kornelia Berghof-Jäger, Sascha Mormann, Ulrich Schotte, Anne Kontek, Nadine Althof, Dietrich Mäde and Reimar Johne

Microorganisms 2020, 8(10), 1460; https://doi.org/10.3390/microorganisms8101460 - 23 Sep 2020

Cited by 2 | Viewed by 2789

Abstract

Background: In the last years, the number of notified hepatitis E cases in humans has continuously increased in Europe. Foodborne infection with the zoonotic hepatitis E virus (HEV) genotype 3 is considered the major cause of this disease. Undercooked liver and raw sausages containing the liver of pigs and wild boar are at high risk of containing HEV. However, so far, no standardized method for the detection of HEV-RNA in pig liver is available. Methods: An international collaborative study on method reproducibility involving 11 laboratories was performed for an HEV-RNA detection method, which consists of steps of sample homogenization, RNA extraction and real-time RT-PCR detection, including a process control. Naturally contaminated pork liver samples containing two different amounts of HEV and a HEV-negative pork liver sample were tested by all laboratories using the method. Results: Valid results were retrieved from 10 laboratories. A specificity of 100% and a sensitivity of 79% were calculated for the method. False negative results were only retrieved from the sample containing very low HEV amounts near the detection limit. Conclusions: The results show that the method is highly specific, sufficiently sensitive and robust for use in different laboratories. The method can, therefore, be applied to routine food control as well as in monitoring studies. Full article

(This article belongs to the Special Issue Enteric Virus Detection: Recent Developments and Application in Food and Waters)

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14 pages, 1363 KiB

Open AccessArticle

Enhancement of the Molecular and Serological Assessment of Hepatitis E Virus in Milk Samples

by Ibrahim M. Sayed, Ahmed R. A. Hammam, Mohamed Salem Elfaruk, Khalid A. Alsaleem, Marwa A. Gaber, Amgad A. Ezzat, Eman H. Salama, Amal A. Elkhawaga and Mohamed A. El-Mokhtar

Microorganisms 2020, 8(8), 1231; https://doi.org/10.3390/microorganisms8081231 - 12 Aug 2020

Cited by 12 | Viewed by 3195

Abstract

Hepatitis E virus (HEV) infection is endemic in developing and developed countries. HEV was reported to be excreted in the milk of ruminants, raising the possibility of transmission of HEV infection through the ingestion of contaminated milk. Therefore, the detection of HEV markers in milk samples becomes pivotal. However, milk includes inhibitory components that affect HEV detection assays. Previously it was reported that dilution of milk matrix improves the performance of HEV molecular assay, however, the dilution of milk samples is not the best strategy especially when the contaminated milk sample has a low HEV load. Therefore, the objective of this study is to compare the effect of extraction procedures on the efficiency of HEV RNA detection in undiluted milk samples. In addition, we assessed the effect of the removal of milk components such as fats and casein on the performance of the molecular and serological assays of HEV. Phosphate buffered saline (PBS) and different milk matrices (such as whole milk, skim milk, and milk serum) were inoculated with different HEV inoculums and subjected to two different extraction procedures. Method A includes manual extraction using spin column-based extraction, while method B includes silica-based automated extraction. Method A was more sensitive than method B in the whole milk and skim milk matrices with a LoD_95% of 300 IU/mL, and virus recovery yield of 47%. While the sensitivity and performance of method B were significantly improved using the milk serum matrix, with LoD_95% of 96 IU/mL. Interestingly, retesting HEV positive milk samples using the high sensitivity assay based on method B extraction and milk serum matrix increased the HEV RNA detection rate to 2-fold. Additionally, the performance of HEV serological assays such as anti-HEV IgG and HEV Ag in the milk samples was improved after the removal of the fat globules from the milk matrix. In conclusion, HEV RNA assay is affected by the components of milk and the extraction procedure. Removal of inhibitory substances, such as fat and casein from the milk sample increased the performance of HEV molecular and serological assays which will be suitable for the low load HEV milk with no further dilutions. Full article

(This article belongs to the Special Issue Enteric Virus Detection: Recent Developments and Application in Food and Waters)

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19 pages, 1920 KiB

Open AccessArticle

Effects of Weather and Environmental Factors on the Seasonal Prevalence of Foodborne Viruses in Irrigation Waters in Gyeonggi Province, Korea

by Zhaoqi Wang, Hansaem Shin, Soontag Jung, Daseul Yeo, Hyunkyung Park, Sangah Shin, Dong Joo Seo, Ki Hwan Park and Changsun Choi

Microorganisms 2020, 8(8), 1224; https://doi.org/10.3390/microorganisms8081224 - 11 Aug 2020

Cited by 12 | Viewed by 2446

Abstract

This study aimed to investigate the prevalence of foodborne viruses in reservoirs (an important resource of irrigation water) and its correlation with environmental and weather factors. From May 2017 to November 2018, we visited ten reservoirs and a river in the Anseong region of South Korea and collected a total of 192 samples in accordance with the environment protection agency guidelines. We recorded the weather factors (temperature, humidity, and accumulated precipitation) and investigated the surrounding environment factors (livestock, fishing site, the catchment area of reservoirs, etc.). Our research results show that from the river and reservoirs, the detection rates of human norovirus GII, adenovirus, rotavirus, human norovirus GI, and astrovirus were 27.1, 10.4, 10.4, 4.16, and 3.1%, respectively. Their viral load ranged from −1.48 to 1.55 log₁₀ genome copies/l. However, hepatitis A virus was not detected in any irrigation water sample. Although no sampling was performed in winter, foodborne viruses and male-specific coliphages were frequently found during spring (40.78%) and autumn (39.47%). Interestingly, the significant correlation between the accumulative precipitation and the number of detected norovirus and adenovirus was confirmed by linear regression analysis. Furthermore, when the accumulative precipitation ranged from 20 to 60 mm, it significantly affected the viral load and prevalence. Among the environmental factors, recreational facilities such as fishing sites and bungalow fishing spots were identified as contamination sources by correlation analysis. Our research results confirmed the correlations between environmental contamination factors in the reservoir and weather factors with the prevalence of foodborne viruses in the reservoir. These facilitates the assessment of potential foodborne virus contamination during crop irrigation. In addition, predictive models including environmental and weather factors should be developed for monitoring and controlling the safety of irrigation waters in reservoirs. Full article

(This article belongs to the Special Issue Enteric Virus Detection: Recent Developments and Application in Food and Waters)

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8 pages, 713 KiB

Open AccessArticle

Assessment of ISO Method 15216 to Quantify Hepatitis E Virus in Bottled Water

by Enric Cuevas-Ferrando, Antonio Martínez-Murcia, Alba Pérez-Cataluña, Gloria Sánchez and Walter Randazzo

Microorganisms 2020, 8(5), 730; https://doi.org/10.3390/microorganisms8050730 - 13 May 2020

Cited by 6 | Viewed by 2812

Abstract

Hepatitis E virus (HEV) is one of the causative agents of water-borne human viral hepatitis and considered in Europe an emerging zoonotic pathogen. Analysis of bottled water through a standard method validated for HEV can contribute towards the risk management of this hazard. Putting some recent reports by the European Food Safety Authority in place, this study aimed to assess the performance of the concentration and extraction procedures described in ISO 15216-1:2017 for norovirus and hepatitis A virus on HEV detection. Following the ISO recommendation, the bottled water samples were spiked using serially diluted HEV fecal suspensions together with mengovirus as process control and concentrated by filtration via positively charged nylon membranes. In order to extract viral RNA from the resulting concentrates, two different methods were compared in this study: The one recommended in the ISO norm, NucliSens^® MiniMag^® system (NS), and an alternative commercially available kit NucleoSpin^®RNA virus kit (MN). Finally, three reverse transcription quantitative PCR (RT-qPCR) assays were used to quantify HEV titers. The evaluated procedures resulted in average HEV recoveries of 14.08 ± 4.90% and 3.58 ± 0.30% for the MN and NS methods, respectively. The limit of detection (LoD_95%) was 1.25 × 10⁴ IU/L for both extraction methods combined with the three RT-qPCR assays tested, with the exception of NS extraction coupled with RT-qPCR1 that showed a LoD_95% of 4.26 × 10³ IU/L. The method characteristics generated in this study support the limited suitability of the ISO 15216-1:2017 concentration procedure coupled with the evaluated RT-qPCR assays for detecting HEV in bottled water. Full article

(This article belongs to the Special Issue Enteric Virus Detection: Recent Developments and Application in Food and Waters)

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21 pages, 5058 KiB

Open AccessArticle

Epidemiological Surveillance of Norovirus and Rotavirus in Sewage (2016–2017) in Valencia (Spain)

by Cristina Santiso-Bellón, Walter Randazzo, Alba Pérez-Cataluña, Susana Vila-Vicent, Roberto Gozalbo-Rovira, Carlos Muñoz, Javier Buesa, Gloria Sanchez and Jesús Rodríguez Díaz

Microorganisms 2020, 8(3), 458; https://doi.org/10.3390/microorganisms8030458 - 24 Mar 2020

Cited by 35 | Viewed by 4521

Abstract

The aim of the present study was to perform the molecular epidemiology of rotaviruses and noroviruses detected in sewage samples from a large wastewater facility from the city of Valencia, Spain. A total of 46 sewage samples were collected over a one-year period (September 2016 to September 2017). Norovirus and rotavirus were detected and quantified by RT-qPCR, genotyped by semi-nested RT-PCR and further characterized by sequencing and phylogenetic analyses. Noroviruses and rotaviruses were widely distributed in sewage samples (69.6% for norovirus GI, 76.0% norovirus GII, and 71.7% rotaviruses) and viral loads varied from 4.33 to 5.75 log PCRU/L for norovirus GI, 4.69 to 6.95 log PCRU/L for norovirus GII, and 4.08 to 6.92 log PCRU/L for rotavirus. Overall, 87.5% (28/32) of GI noroviruses could not be genotyped, 6.25% (2/32) of the samples contained GI.2 genotype, and another 6.25% (2/32) were positive for GI.4 genotype. The most common genotype of GII noroviruses was GII.2 (40%, 14/35), followed by GII.6 (8.6%, 3/35) and GII.17 (5.7%, 2/35) while the remaining GII strains could not be typed (45.7%, 16/35). Rotavirus VP4 genotype P[8] was the only one found in 19 out of 33 rotavirus-positive samples (57.7%). G2 was the most prevalent rotavirus VP7 genotype (15.2%, 5/33) followed by G3, G9, and G12, with two positive samples for each genotype (6.1%, 2/33). In one sample both G1 and G2 genotypes were detected simultaneously (3%). The results presented here show that the surveillance of noroviruses and rotaviruses in sewage is useful for the study of their transmission in the population and their molecular epidemiology. Full article

(This article belongs to the Special Issue Enteric Virus Detection: Recent Developments and Application in Food and Waters)

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