scholarly journals Inactivation of a Human Norovirus Surrogate, Human Norovirus Virus-Like Particles, and Vesicular Stomatitis Virus by Gamma Irradiation

2011 ◽  
Vol 77 (10) ◽  
pp. 3507-3517 ◽  
Author(s):  
Kurtis Feng ◽  
Erin Divers ◽  
Yuanmei Ma ◽  
Jianrong Li
Keyword(s):  
Gamma Irradiation ◽  
Vesicular Stomatitis Virus ◽  
Bacterial Pathogens ◽  
Fresh Produce ◽  
Dose Range ◽  
Murine Norovirus ◽  
Human Norovirus ◽  
Virus Like Particles ◽  
Food Borne ◽  
Vesicular Stomatitis

ABSTRACTGamma irradiation is a nonthermal processing technology that has been used for the preservation of a variety of food products. This technology has been shown to effectively inactivate bacterial pathogens. Currently, the FDA has approved doses of up to 4.0 kGy to control food-borne pathogens in fresh iceberg lettuce and spinach. However, whether this dose range effectively inactivates food-borne viruses is less understood. We have performed a systematic study on the inactivation of a human norovirus surrogate (murine norovirus 1 [MNV-1]), human norovirus virus-like particles (VLPs), and vesicular stomatitis virus (VSV) by gamma irradiation. We demonstrated that MNV-1 and human norovirus VLPs were resistant to gamma irradiation. For MNV-1, only a 1.7- to 2.4-log virus reduction in fresh produce at the dose of 5.6 kGy was observed. However, VSV was more susceptible to gamma irradiation, and a 3.3-log virus reduction at a dose of 5.6 kGy in Dulbecco's modified Eagle medium (DMEM) was achieved. We further demonstrated that gamma irradiation disrupted virion structure and degraded viral proteins and genomic RNA, which resulted in virus inactivation. Using human norovirus VLPs as a model, we provide the first evidence that the capsid of human norovirus has stability similar to that of MNV-1 after exposure to gamma irradiation. Overall, our results suggest that viruses are much more resistant to irradiation than bacterial pathogens. Although gamma irradiation used to eliminate the virus contaminants in fresh produce by the FDA-approved irradiation dose limits seems impractical, this technology may be practical to inactivate viruses for other purposes, such as sterilization of medical equipment.

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.

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 Proton Permeability of HIV Virus like Particles and Vesicular Stomatitis Virus

2014 ◽  
Vol 106 (2) ◽  
pp. 62a
Author(s):  
Pei-I Ku ◽  
Jefferey Hodges ◽  
Michael L. Landesman ◽  
Peter Williams ◽  
Xiaolin Tang ◽  
...  
Keyword(s):  
Vesicular Stomatitis Virus ◽  
Proton Permeability ◽  
Virus Like Particles ◽  
Hiv Virus ◽  
Vesicular Stomatitis

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.

scholarly journals Efficient Production of Human Norovirus-Specific IgY in Egg Yolks by Vaccination of Hens with a Recombinant Vesicular Stomatitis Virus Expressing VP1 Protein

Viruses ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 444 ◽  
Author(s):  
Yang Zhu ◽  
Yuanmei Ma ◽  
Mijia Lu ◽  
Yu Zhang ◽  
Anzhong Li ◽  
...  
Keyword(s):  
Vesicular Stomatitis Virus ◽  
Large Scale ◽  
Passive Immunization ◽  
Therapeutic Interventions ◽  
Efficient Production ◽  
Blood Group Antigens ◽  
Scale Production ◽  
Human Norovirus ◽  
Vp1 Protein ◽  
Vesicular Stomatitis

Human norovirus (HuNoV) is responsible for more than 95% of outbreaks of acute nonbacterial gastroenteritis worldwide. Despite major efforts, there are no vaccines or effective therapeutic interventions against this virus. Chicken immunoglobulin Y (IgY)-based passive immunization has been shown to be an effective strategy to prevent and treat many enteric viral diseases. Here, we developed a highly efficient bioreactor to generate high titers of HuNoV-specific IgY in chicken yolks using a recombinant vesicular stomatitis virus expressing HuNoV capsid protein (rVSV-VP1) as an antigen. We first demonstrated that HuNoV VP1 protein was highly expressed in chicken cells infected by rVSV-VP1. Subsequently, we found that White Leghorn hens immunized intramuscularly with rVSV-VP1 triggered a high level of HuNoV-specific yolk IgY antibodies. The purified yolk IgY was efficiently recognized by HuNoV virus-like particles (VLPs). Importantly, HuNoV-specific IgY efficiently blocked the binding of HuNoV VLPs to all three types (A, B, and O) of histo-blood group antigens (HBGAs), the attachment factors for HuNoV. In addition, the receptor blocking activity of IgY remained stable at temperature below 70 °C and at pH ranging from 4 to 9. Thus, immunization of hens with VSV-VP1 could be a cost-effective and practical strategy for large-scale production of anti-HuNoV IgY antibodies for potential use as prophylactic and therapeutic treatment against HuNoV infection.

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.

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