scholarly journals Evidence of the Internalization of Animal Caliciviruses via the Roots of Growing Strawberry Plants and Dissemination to the Fruit

2015 ◽  
Vol 81 (8) ◽  
pp. 2727-2734 ◽  
Author(s):  
Erin DiCaprio ◽  
Doug Culbertson ◽  
Jianrong Li
Keyword(s):  
Contaminated Soils ◽  
Fresh Produce ◽  
Plaque Assay ◽  
The United States ◽  
Epidemiological Studies ◽  
Murine Norovirus ◽  
Plant Leaves ◽  
Human Norovirus ◽  
Tulane Virus ◽  
Bell Peppers

ABSTRACTHuman norovirus (NoV) is the leading cause of foodborne disease in the United States, and epidemiological studies have shown that fresh produce is one of the major vehicles for the transmission of human NoV. However, the mechanisms of norovirus contamination and persistence in fresh produce are poorly understood. The objective of this study is to determine whether human NoV surrogates, murine norovirus (MNV-1) and Tulane virus (TV), can attach and become internalized and disseminated in strawberries grown in soil. The soil of growing strawberry plants was inoculated with MNV-1 and TV at a level of 108PFU/plant. Leaves and berries were harvested over a 14-day period, and the viral titer was determined by plaque assay. Over the course of the study, 31.6% of the strawberries contained internalized MNV-1, with an average titer of 0.81 ± 0.33 log10PFU/g. In comparison, 37.5% of strawberries were positive for infectious TV, with an average titer of 1.83 ± 0.22 log10PFU/g. A higher percentage (78.7%) of strawberries were positive for TV RNA, with an average titer of 3.15 ± 0.51 log10RNA copies/g as determined by real-time reverse transcriptase quantitative PCR (RT-qPCR). In contrast, no or little virus internalization and dissemination were detected when TV was inoculated into bell peppers grown in soil. Collectively, these data demonstrate (i) virally contaminated soils can lead to the internalization of virus via plant roots and subsequent dissemination to the leaf and fruit portions of growing strawberry plants and (ii) the magnitude of internalization is dependent on the type of virus and plant.

scholarly journals Internalization and Dissemination of Human Norovirus and Animal Caliciviruses in Hydroponically Grown Romaine Lettuce

2012 ◽  
Vol 78 (17) ◽  
pp. 6143-6152 ◽  
Author(s):  
Erin DiCaprio ◽  
Yuanmei Ma ◽  
Anastasia Purgianto ◽  
John Hughes ◽  
Jianrong Li
Keyword(s):  
Real Time ◽  
Fresh Produce ◽  
Plaque Assay ◽  
Feed Water ◽  
Murine Norovirus ◽  
Rt Pcr ◽  
Romaine Lettuce ◽  
Human Norovirus ◽  
Genotype 4 ◽  
Tulane Virus

ABSTRACTFresh produce is a major vehicle for the transmission of human norovirus (NoV) because it is easily contaminated during both pre- and postharvest stages. However, the ecology of human NoV in fresh produce is poorly understood. In this study, we determined whether human NoV and its surrogates can be internalized via roots and disseminated to edible portions of the plant. The roots of romaine lettuce growing in hydroponic feed water were inoculated with 1 × 106RNA copies/ml of a human NoV genogroup II genotype 4 (GII.4) strain or 1 × 106to 2 × 106PFU/ml of animal caliciviruses (Tulane virus [TV] and murine norovirus [MNV-1]), and plants were allowed to grow for 2 weeks. Leaves, shoots, and roots were homogenized, and viral titers and/or RNA copies were determined by plaque assay and/or real-time reverse transcription (RT)-PCR. For human NoV, high levels of viral-genome RNA (105to 106RNA copies/g) were detected in leaves, shoots, and roots at day 1 postinoculation and remained stable over the 14-day study period. For MNV-1 and TV, relatively low levels of infectious virus particles (101to 103PFU/g) were detected in leaves and shoots at days 1 and 2 postinoculation, but virus reached a peak titer (105to 106PFU/g) at day 3 or 7 postinoculation. In addition, human NoV had a rate of internalization comparable with that of TV as determined by real-time RT-PCR, whereas TV was more efficiently internalized than MNV-1 as determined by plaque assay. Taken together, these results demonstrated that human NoV and animal caliciviruses became internalized via roots and efficiently disseminated to the shoots and leaves of the lettuce.

Comparing Human Norovirus Surrogates: Murine Norovirus and Tulane Virus

2013 ◽  
Vol 76 (1) ◽  
pp. 139-143 ◽  
Author(s):  
KIRSTEN A. HIRNEISEN ◽  
KALMIA E. KNIEL
Keyword(s):  
Tap Water ◽  
Plaque Assay ◽  
Heat Treatments ◽  
Viral Infectivity ◽  
Murine Norovirus ◽  
Human Norovirus ◽  
Human Noroviruses ◽  
Ph Values ◽  
Significant Difference ◽  
Tulane Virus

Viral surrogates are widely used by researchers to predict human norovirus behavior. Murine norovirus (MNV) is currently accepted as the best surrogate and is assumed to mimic the survival and inactivation of human noroviruses. Recently, a new calicivirus, the Tulane virus (TV), was discovered, and its potential as a human norovirus surrogate is being explored. This study aimed to compare the behavior of the two potential surrogates under varying treatments of pH (2.0 to 10.0), chlorine (0.2 to 2,000 ppm), heat (50 to 75°C), and survival in tap water at room (20°C) and refrigeration (4°C) temperatures for up to 30 days. Viral infectivity was determined by the plaque assay for both MNV and TV. There was no significant difference between the inactivation of MNV and TV in all heat treatments, and for both MNV and TV survival in tap water at 20°C over 30 days. At 4°C, MNV remained infectious over 30 days at a titer of approximately 5 log PFU/ml, whereas TV titers decreased significantly by 5 days. MNV was more pH stable, as TV titers were reduced significantly at pH 2.0, 9.0, and 10.0, as compared with pH 7.0, whereas MNV titers were only significantly reduced at pH 10.0. After chlorine treatment, there was no significant difference in virus with the exception of at 2 ppm, where TV decreased significantly compared with MNV. Compared with TV, MNV is likely a better surrogate for human noroviruses, as MNV persisted over a wider range of pH values, at 2 ppm of chlorine, and without a loss of titer at 4°C.

scholarly journals Effects of Abiotic and Biotic Stresses on the Internalization and Dissemination of Human Norovirus Surrogates in Growing Romaine Lettuce

2015 ◽  
Vol 81 (14) ◽  
pp. 4791-4800 ◽  
Author(s):  
Erin DiCaprio ◽  
Anastasia Purgianto ◽  
Jianrong Li
Keyword(s):  
Abiotic Stress ◽  
Biotic Stress ◽  
Fresh Produce ◽  
Extreme Weather Events ◽  
Murine Norovirus ◽  
Romaine Lettuce ◽  
Human Norovirus ◽  
Abiotic And Biotic Stresses ◽  
Biotic Stresses ◽  
Tulane Virus

ABSTRACTHuman norovirus (NoV) is the major causative agent of fresh-produce-related outbreaks of gastroenteritis; however, the ecology and persistence of human NoV in produce systems are poorly understood. In this study, the effects of abiotic and biotic stresses on the internalization and dissemination of two human NoV surrogates (murine norovirus 1 [MNV-1] and Tulane virus [TV]) in romaine lettuce were determined. To induce abiotic stress, romaine lettuce was grown under drought and flood conditions that mimic extreme weather events, followed by inoculation of soil with MNV-1 or TV. Independently, lettuce plants were infected with lettuce mosaic virus (LMV) to induce biotic stress, followed by inoculation with TV. Plants were grown for 14 days, and viral titers in harvested tissues were determined by plaque assays. It was found that drought stress significantly decreased the rates of both MNV-1 and TV internalization and dissemination. In contrast, neither flood stress nor biotic stress significantly impacted viral internalization or dissemination. Additionally, the rates of TV internalization and dissemination in soil-grown lettuce were significantly higher than those for MNV-1. Collectively, these results demonstrated that (i) human NoV surrogates can be internalized via roots and disseminated to shoots and leaves of romaine lettuce grown in soil, (ii) abiotic stress (drought) but not biotic stress (LMV infection) affects the rates of viral internalization and dissemination, and (iii) the type of virus affects the efficiency of internalization and dissemination. This study also highlights the need to develop effective measures to eliminate internalized viruses in fresh produce.

Application of chitosan microparticles against human norovirus

2021 ◽  
Author(s):  
Candace Barnes ◽  
Rebecca Barber ◽  
Keith R. Schneider ◽  
Michelle D. Danyluk ◽  
Anita C. Wright ◽  
...  
Keyword(s):  
Environmental Water ◽  
The United States ◽  
Virus Infectivity ◽  
Log10 Reduction ◽  
Human Norovirus ◽  
Virus Particles ◽  
Chitosan Microparticles ◽  
Natural Treatment ◽  
Fecal Suspension ◽  
Tulane Virus

Human norovirus (HuNoV) is the leading causative agent of foodborne outbreaks and is associated with the second most prevalent cause of waterborne infections in the United States. The goal of this research was to investigate the antiviral activity of chitosan microparticles (CM) against HuNoV GII.4 Sydney and its cultivable surrogate, Tulane virus (TuV), in suspensions mimicking fecally-contaminated water. CM was prepared by crosslinking chitosan molecules with sodium sulfate, and then its anti-noroviral activity was assessed using infectivity assay on TuV and RT-qPCR on TuV and HuNoV. A 3% CM suspension in PBS (pH 7.2) showed binding to TuV particles but with a negligible impact on virus infectivity (p>0.05). TuV and HuNoV suspended in fecal suspensions showed a 1.5-log10 reduction in genomic copies per ml following a 10-min contact time (p<0.05). Despite the negligible impact on viral infectivity, CM moderately binds to virus particles and helps purify environmental water by removing infectious virus particles. In this study, TuV served as a suitable surrogate for HuNoV by showing a similar log10 reduction in fecal suspension. Overall, the outcomes of thisresearch highlight the potential application of CM as a novel, natural treatment to minimize the spread of water-transmitted viral pathogens.

scholarly journals Comprehensive Comparison of Cultivable Norovirus Surrogates in Response to Different Inactivation and Disinfection Treatments

2014 ◽  
Vol 80 (18) ◽  
pp. 5743-5751 ◽  
Author(s):  
Theresa Cromeans ◽  
Geun Woo Park ◽  
Veronica Costantini ◽  
David Lee ◽  
Qiuhong Wang ◽  
...  
Keyword(s):  
Culture Method ◽  
Aichi Virus ◽  
Murine Norovirus ◽  
Human Norovirus ◽  
The Public ◽  
Test Parameters ◽  
Reverse Transcription Quantitative Pcr ◽  
Tulane Virus ◽  
Comprehensive Comparison ◽  
Identical Test

ABSTRACTHuman norovirus is the leading cause of epidemic and sporadic acute gastroenteritis. Since no cell culture method for human norovirus exists, cultivable surrogate viruses (CSV), including feline calicivirus (FCV), murine norovirus (MNV), porcine enteric calicivirus (PEC), and Tulane virus (TuV), have been used to study responses to inactivation and disinfection methods. We compared the levels of reduction in infectivities of CSV and Aichi virus (AiV) after exposure to extreme pHs, 56°C heating, alcohols, chlorine on surfaces, and high hydrostatic pressure (HHP), using the same matrix and identical test parameters for all viruses, as well as the reduction of human norovirus RNA levels under these conditions. At pH 2, FCV was inactivated by 6 log10units, whereas MNV, TuV, and AiV were resistant. All CSV were completely inactivated at 56°C within 20 min. MNV was inactivated 5 log10units by alcohols, in contrast to 2 and 3 log10units for FCV and PEC, respectively. TuV and AiV were relatively insensitive to alcohols. FCV was reduced 5 log10units by 1,000 ppm chlorine, in contrast to 1 log10unit for the other CSV. All CSV except FCV, when dried on stainless steel surfaces, were insensitive to 200 ppm chlorine. HHP completely inactivated FCV, MNV, and PEC at ≥300 MPa, and TuV at 600 MPa, while AiV was completely resistant to HHP up to 800 MPa. By reverse transcription-quantitative PCR (RT-qPCR), genogroup I (GI) noroviruses were more sensitive than GII noroviruses to alcohols, chlorine, and HHP. Although inactivation profiles were variable for each treatment, TuV and MNV were the most resistant CSV overall and therefore are the best candidates for studying the public health outcomes of norovirus infections.

Inactivation of Human Norovirus and Its Surrogates on Alfalfa Seeds by Aqueous Ozone

2015 ◽  
Vol 78 (8) ◽  
pp. 1586-1591 ◽  
Author(s):  
QING WANG ◽  
SARAH MARKLAND ◽  
KALMIA E. KNIEL
Keyword(s):  
Deionized Water ◽  
Murine Norovirus ◽  
Food Matrix ◽  
Human Norovirus ◽  
Viral Genomes ◽  
Tulane Virus ◽  
Foodborne Outbreaks ◽  
Alfalfa Sprouts ◽  
All Treatment ◽  
Alfalfa Seeds

Alfalfa sprouts have been associated with numerous foodborne outbreaks. Previous studies investigated the effectiveness of aqueous ozone on bacterially contaminated seeds, yet little is known about the response of human norovirus (huNoV). This study assessed aqueous ozone for the disinfection of alfalfa seeds contaminated with huNoV and its surrogates. The inactivation of viruses without a food matrix was also investigated. Alfalfa seeds were inoculated with huNoV genogroup II, Tulane virus (TV), and murine norovirus (MNV); viruses alone or inoculated on seeds were treated in deionized water containing 6.25 ppm of aqueous ozone with agitation at 22°C for 0.5, 1, 5, 15, or 30 min. The data showed that aqueous ozone resulted in reductions of MNV and TV infectivity from 1.66 ± 1.11 to 5.60 ± 1.11 log PFU/g seeds; for all treatment times, significantly higher reductions were observed for MNV (P < 0.05). Viral genomes were relatively resistant, with a reduction of 1.50 ± 0.14 to 3.00 ± 0.14 log genomic copies/g seeds; the reduction of TV inoculated in water was similar to that of huNoV, whereas MNV had significantly greater reductions in genomic copies (P < 0.05). Similar trends were observed in ozone-treated viruses alone, with significantly higher levels of inactivation (P < 0.05), especially with reduced levels of infectivity for MNV and TV. The significant inactivation by aqueous ozone indicates that ozone may be a plausible substitute for chlorine as an alternative treatment for seeds. The behavior of TV was similar to that of huNoV, which makes it a promising surrogate for these types of scenarios.

scholarly journals Interactions between Human Norovirus Surrogates and Acanthamoeba spp.

2015 ◽  
Vol 81 (12) ◽  
pp. 4005-4013 ◽  
Author(s):  
Tun-Yun Hsueh ◽  
Kristen E. Gibson
Keyword(s):  
Fresh Produce ◽  
Disease Outbreaks ◽  
The United States ◽  
Feline Calicivirus ◽  
Murine Norovirus ◽  
Virus Inoculation ◽  
Free Living ◽  
Content Type ◽  
Food Borne

ABSTRACTHuman noroviruses (HuNoVs) are the most common cause of food-borne disease outbreaks, as well as virus-related waterborne disease outbreaks in the United States. Here, we hypothesize that common free-living amoebae (FLA)—ubiquitous in the environment, known to interact with pathogens, and frequently isolated from water and fresh produce—could potentially act as reservoirs of HuNoV and facilitate the environmental transmission of HuNoVs. To investigate FLA as reservoirs for HuNoV, the interactions between twoAcanthamoebaspecies,A. castellaniiandA. polyphaga, as well as two HuNoV surrogates, murine norovirus type 1 (MNV-1) and feline calicivirus (FCV), were evaluated. The results showed that after 1 h of amoeba-virus incubation at 25°C, 490 and 337 PFU of MNV-1/ml were recovered fromA. castellaniiandA. polyphaga, respectively, while only few or no FCVs were detected. In addition, prolonged interaction of MNV-1 with amoebae was investigated for a period of 8 days, and MNV-1 was demonstrated to remain stable at around 200 PFU/ml from day 2 to day 8 after virus inoculation inA. castellanii. Moreover, after a complete amoeba life cycle (i.e., encystment and excystment), infectious viruses could still be detected. To determine the location of virus associated with amoebae, immunofluorescence experiments were performed and showed MNV-1 transitioning from the amoeba surface to inside the amoeba over a 24-h period. These results are significant to the understanding of how HuNoVs may interact with other microorganisms in the environment in order to aid in its persistence and survival, as well as potential transmission in water and to vulnerable food products such as fresh produce.

scholarly journals Internalization and dissemination of human norovirus and Tulane virus in fresh produce is plant dependent

2018 ◽  
Vol 69 ◽  
pp. 25-32 ◽  
Author(s):  
Zhihong Yang ◽  
Heather Chambers ◽  
Erin DiCaprio ◽  
Gary Gao ◽  
Jianrong Li
Keyword(s):  
Fresh Produce ◽  
Human Norovirus ◽  
Tulane Virus

Evaluation of Murine Norovirus Persistence in Environments Relevant to Food Production and Processing

2011 ◽  
Vol 74 (11) ◽  
pp. 1847-1851 ◽  
Author(s):  
S. FALLAHI ◽  
K. MATTISON
Keyword(s):  
Stainless Steel ◽  
Food Production ◽  
Room Temperature ◽  
Potable Water ◽  
Plaque Assay ◽  
The Other ◽  
Murine Norovirus ◽  
Human Norovirus ◽  
Log Reduction ◽  
Virus Survival

Human norovirus (NoV) causes outbreaks of acute gastroenteritis associated with many ready-to-eat foods, including fresh produce. Effective inactivation procedures must consider virus survival under conditions of produce production and processing. This study aimed to investigate the persistence of NoV in a variety of environments, using murine NoV (MNV) as a surrogate for NoV. MNV was incubated for up to 42 days at room temperature on stainless steel disks, on lettuce, on soil, and in potable water and titers determined by plaque assay. A 1-log reduction of MNV infectivity was observed after 29 days in water, 4 days on lettuce, 12 days on soil, and 15 days on stainless steel disks. MNV survived longer in water than in any of the other environments, indicating that drying may contribute to NoV inactivation. MNV genomes were not significantly reduced for up to 42 days, suggesting that genomic detection is not a reliable indicator of viability. Overall, our findings provide valuable information regarding the potential for NoV transmission in the food supply.

Enhanced Removal of Norovirus Surrogates, Murine Norovirus and Tulane Virus, from Aqueous Systems by Zero-Valent Iron

2018 ◽  
Vol 81 (9) ◽  
pp. 1432-1438 ◽  
Author(s):  
ADRIENNE E. H. SHEARER ◽  
KALMIA E. KNIEL
Keyword(s):  
Field Studies ◽  
Zero Valent Iron ◽  
Water Remediation ◽  
Murine Norovirus ◽  
Human Norovirus ◽  
Elution Buffer ◽  
Biological Contaminants ◽  
Tulane Virus ◽  
Viral Contaminants ◽  
Genomic Material

ABSTRACTViral contamination can compromise the safety of water utilized for direct consumption, produce irrigation, and postharvest washing of produce. Zero-valent iron (ZVI) is used commercially for chemical remediation of water and has been demonstrated to remove some biological contaminants from water in laboratory and field studies. This study investigated the efficacy of ZVI to remove human norovirus surrogates, Tulane virus (TV) and murine norovirus (MNV), from water and to characterize the reversibility and nature of viral association with ZVI. Genomic material of TV and MNV recovered from the effluent of inoculated water treatment columns containing a 1:1 mixture of ZVI and sand was 2 and 3 log, respectively, less than that recovered from the effluent of treatment columns containing only sand. Elution buffers (citrate buffers, pH 4 and 7, and virus elution buffer, pH 9.5, with and without added 1 M NaCl) did not increase recovery of infectious TV and MNV from ZVI as compared with elution with water alone. TV-inoculated lettuce washed with water in the presence of ZVI yielded 1.5 to 2 log fewer infectious TV from washwater as compared with lettuce washed with water alone or in the presence of sand. These data demonstrate the enhanced removal of human norovirus surrogates, TV and MNV, from water by ZVI and provide indications that unrecovered viruses are not readily disassociated from ZVI by buffers of various pH and ionic strength. These findings warrant further investigation into larger-scale simulations of water remediation of viral contaminants for potential application in the treatment of water used for drinking, irrigation, and food processing.

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