Environmental surveillance of norovirus and hepatitis A virus in raw oysters at seafood retails in Sinaloa, Mexico: detection of GII.P13 norovirus genotype

2019 ◽  
Author(s):  
Claudia Villicaña ◽  
Luis Amarillas ◽  
América Cota-Álvarez ◽  
Josefina León-Félix ◽  
Bruno Gómez-Gil
Keyword(s):  
Genetic Diversity ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Binary Logistic Regression ◽  
Significant Negative Correlation ◽  
Human Infection ◽  
Fecal Coliforms ◽  
Environmental Surveillance ◽  
Period 2 ◽  
Food Borne

Abstract Background Norovirus (NoV) and hepatitis A virus (HAV) have emerged as the leading agents of non-bacterial acute gastroenteritis and hepatitis, respectively, primarily associated to food-borne outbreaks followed by the consumption of seafood. In Mexico, little is known about the molecular epidemiology of NoV and HAV, and a few studies have reported their presence in aquatic environments. In Sinaloa, the pleasure oyster (Crassostrea cortiziensis, Hertlein) is one of the main species consumed at seafood retails; however, information about the presence and genetic diversity of NoV and HAV is lacking. The aim of the present study was to investigate the prevalence and molecular diversity of NoV and HAV in raw pleasure oysters expended at seafood retails in Sinaloa, Mexico. Methods A total of 68 samples were collected at several seafood retails at four periods: Period 1 (October-November, 2010); Period 2 (December 2010-January 2011); Period 3 (March-April, 2011); and Period 4 (May, 2011). These oyster samples were tested for the presence of NoV and HAV by retrotranscription (RT)-nested PCR and RT-PCR, respectively. Enumeration of fecal coliforms was also conducted. In addition, an analysis of Binary Logistic Regression (BLR) was performed to determine a possible correlation of these enteric viruses with the presence of fecal coliforms and environmental temperature. Results Overall, NoV was detected in 57.3% of the 68 samples showing the highest occurrence (11/13) during Period 1 (October-November, 2010). No HAV-positive samples were detected. Fecal coliforms were more frequently detected (16/20) on Period 4 (May, 2011). A significant negative correlation between NoV and fecal coliforms was observed. A total of 28 sequences were obtained from NoV amplicons and surprisingly, phylogenetic analysis showed that all NoV sequences obtained from oysters belonged to GII.P13 genotype. Conclusions The results indicated that raw oysters expended at seafood retails are potential sources of human infection due to the presence of NoV, which interestingly were represented only by GII.P13 genotype. This is the first report confirming the presence of GII.P13 in Mexico, which may contribute with the better understanding of NoV genetic diversity and epidemiology.

scholarly journals Contamination of Foods by Food Handlers: Experiments on Hepatitis A Virus Transfer to Food and Its Interruption

2000 ◽  
Vol 66 (7) ◽  
pp. 2759-2763 ◽  
Author(s):  
S. Bidawid ◽  
J. M. Farber ◽  
S. A. Sattar
Keyword(s):  
Hepatitis A ◽  
Infectious Virus ◽  
Hepatitis A Virus ◽  
Infectious Hepatitis ◽  
Topical Agent ◽  
Food Handlers ◽  
Virus Removal ◽  
Virus Transfer ◽  
Food Borne ◽  
Water Rinsing

ABSTRACT Hepatitis A virus (HAV) is an important pathogen which has been responsible for many food-borne outbreaks. HAV-excreting food handlers, especially those with poor hygienic practices, can contaminate the foods which they handle. Consumption of such foods without further processing has been known to result in cases of infectious hepatitis. Since quantitative data on virus transfer during contact of hands with foods is not available, we investigated the transfer of HAV from artificially contaminated fingerpads of adult volunteers to pieces of fresh lettuce. Touching the lettuce with artificially contaminated fingerpads for 10 s at a pressure of 0.2 to 0.4 kg/cm2resulted in transfer of 9.2% � 0.9% of the infectious virus. The pretreatments tested to interrupt virus transfer from contaminated fingerpads included (i) hard-water rinsing and towel drying, (ii) application of a domestic or commercial topical agent followed by water rinsing and towel drying, and (iii) exposure to a hand gel containing 62% ethanol or 75% liquid ethanol without water rinsing or towel drying. When the fingerpads were treated with the topical agents or alcohol before the lettuce was touched, the amount of infectious virus transferred to lettuce was reduced from 9.2% to between 0.3 and 0.6% (depending on the topical agent used), which was a reduction in virus transfer of up to 30-fold. Surprisingly, no virus transfer to lettuce was detected when the fingerpads were rinsed with water alone before the lettuce was touched. However, additional experiments with water rinsing in which smaller volumes of water were used (1 ml instead of 15 ml) showed that the rate of virus transfer to lettuce was 0.3% � 0.1%. The variability in virus transfer rates following water rinsing may indicate that the volume of water at least in part influences virus removal from the fingerpads differently, a possibility which should be investigated further. This study provided novel information concerning the rate of virus transfer to foods and a model for investigating the transfer of viral and other food-borne pathogens from contaminated hands to foods, as well as techniques for interrupting such transfer to improve food safety.

scholarly journals Molecular epidemiology and genetic diversity of duck hepatitis A virus type 3 in Shandong province of China, 2012–2014

2017 ◽  
Vol 61 (04) ◽  
pp. 463-472 ◽  
Author(s):  
R. ZHANG ◽  
L. XIA ◽  
J. CHEN ◽  
Y. GONG ◽  
L. ZHANG ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Molecular Epidemiology ◽  
Virus Type ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Shandong Province ◽  
Duck Hepatitis ◽  
Type 3

scholarly journals Detection of both Hepatitis A Virus and Norwalk-Like Virus in Imported Clams Associated with Food-Borne Illness

2002 ◽  
Vol 68 (8) ◽  
pp. 3914-3918 ◽  
Author(s):  
David H. Kingsley ◽  
Gloria K. Meade ◽  
Gary P. Richards
Keyword(s):  
Hepatitis A ◽  
Magnetic Beads ◽  
Hepatitis A Virus ◽  
New York State ◽  
Rna Extraction ◽  
The United States ◽  
Fecal Coliforms ◽  
Most Probable Number ◽  
Genotype I ◽  
Probable Number

ABSTRACT Hepatitis A virus (HAV) and Norwalk-like virus (NLV) were detected by reverse transcription-PCR in clams imported into the United States from China. An epidemiological investigation showed that these clams were associated with five cases of Norwalk-like gastroenteritis in New York State in August 2000 (Food and Drug Administration Import Alert 16-50). They were labeled “cooked” but appeared raw. Viral RNA extraction was performed by using dissected digestive tissues rather than whole shellfish meats; this was followed by glycine buffer elution, polyethylene glycol precipitation, Tri-Reagent treatment, and purification of poly(A) RNA with magnetic beads coupled to poly(dT) oligonucleotides. We identified HAV and NLV as genotype I and genogroup II strains, respectively. Both viruses have high levels of homology to Asian strains. An analysis of fecal coliforms revealed a most-probable number of 93,000/100 g of clam meat, which is approximately 300-fold higher than the hygienic standard for shellfish meats.

scholarly journals Development of Methods To Detect “Norwalk-Like Viruses” (NLVs) and Hepatitis A Virus in Delicatessen Foods: Application to a Food-Borne NLV Outbreak

2000 ◽  
Vol 66 (1) ◽  
pp. 213-218 ◽  
Author(s):  
Kellogg J. Schwab ◽  
Frederick H. Neill ◽  
Rebecca L. Fankhauser ◽  
Nicholas A. Daniels ◽  
Stephan S. Monroe ◽  
...  
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Internal Standard ◽  
Food Samples ◽  
Viral Rna ◽  
Norwalk Virus ◽  
Specific Primers ◽  
Food Borne ◽  
Common Causes ◽  
Food Borne Illness

ABSTRACT “Norwalk-like viruses” (NLVs) and hepatitis A virus (HAV) are the most common causes of virus-mediated food-borne illness. Epidemiological investigations of outbreaks associated with these viruses have been hindered by the lack of available methods for the detection of NLVs and HAV in foodstuffs. Although reverse transcription (RT)-PCR methods have been useful in detecting NLVs and HAV in bivalve mollusks implicated in outbreaks, to date such methods have not been available for other foods. To address this need, we developed a method to detect NLVs and HAV recovered from food samples. The method involves washing of food samples with a guanidinium-phenol-based reagent, extraction with chloroform, and precipitation in isopropanol. Recovered viral RNA is amplified with HAV- or NLV-specific primers in RT-PCRs, using a viral RNA internal standard control to identify potential sample inhibition. By this method, 10 to 100 PCR units (estimated to be equivalent to 102 to 103 viral genome copies) of HAV and Norwalk virus seeded onto ham, turkey, and roast beef were detected. The method was applied to food samples implicated in an NLV-associated outbreak at a university cafeteria. Sliced deli ham was positive for a genogroup II NLV as determined by using both polymerase- and capsid-specific primers and probes. Sequence analysis of the PCR-amplified capsid region of the genome indicated that the sequence was identical to the sequence from virus detected in the stools of ill students. The developed method is rapid, simple, and efficient.

scholarly journals Genetic diversity, haplotype analysis, and risk factor assessment of hepatitis A virus isolates from the West Bank, Palestine during the period between 2014 and 2016

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0240339
Author(s):  
Kamal Dumaidi ◽  
Hayah Qaraqe ◽  
Amer Al-Jawabreh ◽  
Rasmi Abu-Helu ◽  
Fekri Samarah ◽  
...  
Keyword(s):  
Risk Factors ◽  
Genetic Diversity ◽  
Phylogenetic Analysis ◽  
Hepatitis A ◽  
Nucleotide Diversity ◽  
Hepatitis A Virus ◽  
West Bank ◽  
Haplotype Network ◽  
Hand Washing ◽  
The West

Background Hepatitis A virus (HAV) infection is one of the major causes of acute viral hepatitis. HAV genotypes and its genetic diversity is rarely investigated in our region as well as worldwide. Aims The aims of the present study were to determine the HAV genotypes and its risk factors and to investigate the genetic diversity of the HAV isolates in the West Bank, Palestine. Study design A cohort of 161 clinically and laboratory-confirmed HAV (IgM-positive) cases and 170 apparently healthy controls from all the districts of the West Bank, Palestine during the period of 2014 to 2016 were tested for HAV infection using IgM antibodies, RT-PCR and sequence analysis of the VP3/VP1 junction region of the HAV genome. Phylogenetic analysis, genetic diversity and haplotypes analysis were used to characterize the VP3/VP1 sequences. Results All the 34 sequences of the HAV were found to be of HAV-IB sub-genotype. The phylogenetic analysis showed four main clusters with cluster III exclusively consisting of 18 Palestinian isolates (18/23-78%), but with weak bootstrap values. A high haplotype diversity (Hd) and low nucleotide diversity (π) were observed. Cluster III showed high number of haplotypes (h = 8), but low haplotype (gene) diversity (Hd = 0.69). A total of 28 active haplotypes with some consisting of more than one sequence were observed using haplotype network analysis. The Palestinian haplotypes are characterized by closely related viral haplotypes with one SNV away from each other which ran parallel to cluster III in the phylogenetic tree. A smaller Palestinian haplotype (4 isolates) was three SNVs away from the major haplotype cluster (n = 10) and closer to others haplotypes from Iran, Spain, and South Africa. Young age, low level of parent’s education, infrequent hand washing before meals, and drinking of un-treated water were considered the major HAV risk factors in the present study. Conclusion Haplotype network analysis revealed haplotype variation among the HAV Palestinian sequences despite low genetic variation and nucleotide diversity. In addition, this study reconfirmed that age and parent’s level of education as HAV risk factors, while hand washing and treating drinking water as protective factors.

scholarly journals Genetic Diversity of Hepatitis A Virus in China: VP3-VP1-2A Genes and Evidence of Quasispecies Distribution in the Isolates

PLoS ONE ◽  
2013 ◽  
Vol 8 (9) ◽  
pp. e74752 ◽  
Author(s):  
Hao Wang ◽  
Huihui Zheng ◽  
Jingyuan Cao ◽  
Wenting Zhou ◽  
Yao Yi ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Hepatitis A ◽  
Hepatitis A Virus

scholarly journals Survival of Murine Norovirus, Tulane Virus, and Hepatitis A Virus on Alfalfa Seeds and Sprouts during Storage and Germination

2013 ◽  
Vol 79 (22) ◽  
pp. 7021-7027 ◽  
Author(s):  
Qing Wang ◽  
Kirsten A. Hirneisen ◽  
Sarah M. Markland ◽  
Kalmia E. Kniel
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Storage Period ◽  
Testing Time ◽  
Murine Norovirus ◽  
Food Borne ◽  
Time Period ◽  
Tulane Virus ◽  
Viral Etiologies ◽  
Alfalfa Seeds

ABSTRACTHuman norovirus (huNoV) and hepatitis A virus (HAV) have been involved in several produce-associated outbreaks and identified as major food-borne viral etiologies. In this study, the survival of huNoV surrogates (murine norovirus [MNV] and Tulane virus [TV]) and HAV was investigated on alfalfa seeds during storage and postgermination. Alfalfa seeds were inoculated with MNV, TV, or HAV with titers of 6.46 ± 0.06 log PFU/g, 3.87 ± 0.38 log PFU/g, or 7.01 ± 0.07 log 50% tissue culture infectious doses (TCID50)/g, respectively. Inoculated seeds were stored for up to 50 days at 22°C and sampled during that storage period on days 0, 2, 5, 10, and 15. Following storage, virus presence was monitored over a 1-week germination period. Viruses remained infectious after 50 days, with titers of 1.61 ± 0.19 log PFU/g, 0.85 ± 0.21 log PFU/g, and 3.43 ± 0.21 log TCID50/g for MNV, TV, and HAV, respectively. HAV demonstrated greater persistence than MNV and TV, without a statistically significant reduction over 20 days (<1 log TCID50/g); however, relatively high levels of genomic copies of all viruses persisted over the testing time period. Low titers of viruses were found on sprouts and were located in all tissues as well as in sprout-spent water sampled on days 1, 3, and 6 following seed planting. Results revealed the persistence of viruses in seeds for a prolonged period of time, and perhaps of greater importance these data suggest the ease of which virus may transfer from seeds to sprouts and spent water during germination. These findings highlight the importance of sanitation and prevention procedures before and during germination.

scholarly journals New trends in food- and waterborne viral outbreaks

2014 ◽  
Vol 66 (1) ◽  
pp. 1-9 ◽  
Author(s):  
Dragoslava Radin
Keyword(s):  
Food Production ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Contaminated Water ◽  
Food Handlers ◽  
Emerging Viruses ◽  
Current Trends ◽  
Food Borne Pathogens ◽  
Food Borne ◽  
Public Health Challenges

Current trends in food- and waterborne viral diseases have been reviewed. Awareness and surveillance of viral food and waterborne pathogens is generally not sufficient, with emphasis placed on noroviruses, hepatitis A virus, rotaviruses and newly emerging viruses. In addition, previously unknown food-borne pathogens, many of which are zoonotic, are constantly emerging. Food can be contaminated with a virus either at the source via contaminated water, or at the point of service by infected food handlers. Viruses can spread by water, direct person-to-person contact, airborne droplets or vomit, and they can persist in the environment as a source of continuing infection despite disinfection efforts. Food production and supply practices change, and food-borne pathogens seem able to exploit novel opportunities, for example fresh produce, and generate new food safety and public health challenges.

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