Murine Norovirus: Propagation, Quantification, and Genetic Manipulation

Hwang, Seungmin; Alhatlani, Bader Y.; Arias, Armando; Caddy, Sarah; Christodoulou, Constantina; Cunha, Juliana Bragazzi; Emmott, Edward; Gonzalez-Hernandez, Marta J.; Kolawole, Abimbola O.; Jia, Lianshun; Rippinger, Christine M.; Sorgeloos, Frédéric; Thorne, Lucy; Vashist, Surender; Goodfellow, Ian; Wobus, Christiane E.

doi:10.1002/9780471729259.mc15k02s33

Cited by 86 publications

(80 citation statements)

References 54 publications

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“…One major difference in the virus-infected cell lysates of FCV, SaV, MNV, and TV was the concentration of FBS used in their propagation. We propagated these viruses as reported previously for MNV in 10% FBS, TV in 5%, FCV in 2%, and SaV in 0% FBS (11,12,28). However, other studies have used various concentrations of FBS for propagation of these viruses.…”

Section: Discussionmentioning

confidence: 99%

“…As reported previously, SaV and FCV were cultured in minimum essential medium (MEM; Life Technologies, Grand Island, NY) supplemented with nonessential amino acids (1%) without or with 2% fetal bovine serum (FBS), respectively (11). TV and MNV were cultured in M199 and Dulbecco's modified Eagle's medium (DMEM) with high glucose (Life Technologies) supplemented with 5% and 10% FBS, respectively (12,28). In addition, all culture media used for cell maintenance and virus generation were supplemented with 1% HEPES and 1% antibiotic-antimycotic cocktail (Invitrogen, Carlsbad, CA, USA).…”

Section: Propagation Of Virusesmentioning

confidence: 99%

“…Cell culture propagation of SaV (Cowden strain), FCV (F9 strain), MNV (S7 strain), and TV in an LLC porcine kidney cell line (LLC-PK1; ATCC CL-101), the Crandell-Rees feline kidney cell line (CRFK; ATCC CCL-94), a mouse leukemic macrophage cell line (RAW 264.7; ATCC TIB-71), and an LLC monkey kidney cell line (LLC-MK2; ATCC CCL-7) was done as described previously (11,12,28). The MNV S7 strain was generously provided by Yukinobu Tohya, Department of Veterinary Medicine, Nihon University, Japan (29).…”

Section: Propagation Of Virusesmentioning

confidence: 99%

See 2 more Smart Citations

Feline Calicivirus, Murine Norovirus, Porcine Sapovirus, and Tulane Virus Survival on Postharvest Lettuce

Esseili

Saif

Farkas

et al. 2015

Appl Environ Microbiol

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bHuman norovirus (HuNoV) is the leading cause of foodborne illnesses, with an increasing number of outbreaks associated with leafy greens. Because HuNoV cannot be routinely cultured, culturable feline calicivirus (FCV), murine norovirus (MNV), porcine sapovirus (SaV), and Tulane virus (TV) have been used as surrogates. These viruses are generated in different cell lines as infected cell lysates, which may differentially affect their stability. Our objective was to uniformly compare the survival of these viruses on postharvest lettuce while evaluating the effects of cell lysates on their survival. Viruses were semipurified from cell lysates by ultrafiltration or ultracentrifugation followed by resuspension in sterile water. Virus survival was examined before and after semipurification: in suspension at room temperature (RT) until day 28 and on lettuce leaves stored at RT for 3 days or at 4°C for 7 and 14 days. In suspension, both methods significantly enhanced the survival of all viruses. On lettuce, the survival of MNV in cell lysates was similar to that in water, under all storage conditions. In contrast, the survival of FCV, SaV, and TV was differentially enhanced, under different storage conditions, by removing cell lysates. Following semipurification, viruses showed similar persistence to each other on lettuce stored under all conditions, with the exception of ultracentrifugation-purified FCV, which showed a higher inactivation rate than MNV at 4°C for 14 days. In conclusion, the presence of cell lysates in viral suspensions underestimated the survivability of these surrogate viruses, while viral semipurification revealed similar survivabilities on postharvest lettuce leaves. Human noroviruses (HuNoVs) are the leading cause of acute viral gastroenteritis in the United States (1). These viruses are 28 to 35 nm in diameter, nonenveloped, single-stranded RNA viruses that are members of the Caliciviridae family. An increasing number of HuNoV outbreaks have been associated with leafy greens (2). For example, food was the primary vehicle of transmission in 1,008 HuNoV outbreaks reported between 2009 and 2012 in the United States, and leafy greens were the primary food associated with these outbreaks (1). Therefore, there is a need to understand the occurrence and survival/persistence of HuNoV on leafy greens. However, assessing the survival of HuNoVs on leafy greens is hampered by the fact that these viruses are still refractory to routine growth in cell culture; hence, their infectivity cannot be readily quantified (3, 4). Therefore, surrogate culturable viruses are often used as proxies to investigate HuNoV transmission routes, disinfection, and survival in the environment.Among members of the Caliciviridae family, feline calicivirus (FCV) has been the most widely used HuNoV surrogate since the 1990s (5). In 2004, murine norovirus (MNV), a virus genetically more closely related to HuNoV, was propagated in cell culture (6) and, since then, it has been widely used as a HuNoV surrogate (7). In 2004, bile acids in i...

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Section: Discussionmentioning

confidence: 99%

Section: Propagation Of Virusesmentioning

confidence: 99%

Section: Propagation Of Virusesmentioning

confidence: 99%

See 1 more Smart Citation

Feline Calicivirus, Murine Norovirus, Porcine Sapovirus, and Tulane Virus Survival on Postharvest Lettuce

Esseili

Saif

Farkas

et al. 2015

Appl Environ Microbiol

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“…For light-sensitive MNV, neutral red (NR)-containing CR3 stocks were generated as described previously (18,28). For high-titer infections, virus stocks were concentrated as described previously (29). All light-sensitive virus preparations displayed about 4-log to 6-log reductions in viral titers upon the exposure of regular and concentrated virus stocks, respectively, to light.…”

Section: Methodsmentioning

confidence: 99%

Oral Norovirus Infection Is Blocked in Mice Lacking Peyer's Patches and Mature M Cells

Kolawole

Gonzalez-Hernandez

Turula

et al. 2016

J Virol

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“…Cell culture propagation of MNV, SaV, and TV was performed as described previously (17,25,26) using the following cell lines: a mouse leukemic macrophage cell line (RAW 264.7, ATCC TIB-71), an LLC-porcine kidney cell line (LLC-PK1, ATCC CL-101), and an LLC-monkey kidney cell line (LLC-MK2, ATCC CCL-7), respectively. Briefly, SaV was propagated in minimum essential medium (MEM; Life Technologies, Grand Island, NY) without fetal bovine serum (FBS), while TV and MNV were propagated in M199 cell culture medium and highglucose Dulbecco modified Eagle medium (Life Technologies) containing 5 and 10% FBS (Thermo Scientific, Rockford, IL), respectively.…”

Section: Preparation Of Virusesmentioning

confidence: 99%

Abiotic Stress and Phyllosphere Bacteria Influence the Survival of Human Norovirus and Its Surrogates on Preharvest Leafy Greens

Esseili

Gao

Tegtmeier

et al. 2016

Appl Environ Microbiol

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Foodborne outbreaks of human noroviruses (HuNoVs) are frequently associated with leafy greens. Because there is no effective method to eliminate HuNoV from postharvest leafy greens, understanding virus survival under preharvest conditions is crucial. The objective of this study was to evaluate the survival of HuNoV and its surrogate viruses, murine norovirus (MNV), porcine sapovirus (SaV), and Tulane virus (TV), on preharvest lettuce and spinach that were subjected to abiotic stress (physical damage, heat, or flood). We also examined the bacteria culturable from the phyllosphere in response to abiotic stress and in relation to viral persistence. Mature plants were subjected to stressors 2 days prior to inoculation of the viruses on leaves. We quantified the viral RNA, determined the infectivity of the surrogates, and performed bacterial counts on postinoculation days (PIDs) 0, 1, 7, and 14. For both plant types, time exerted significant effects on HuNoV, MNV, SaV, and TV RNA titers, with greater effects being seen for the surrogates. Infectious surrogate viruses were undetectable on PID 14. Only physical damage on PID 14 significantly enhanced HuNoV RNA persistence on lettuce, while the three stressors differentially enhanced the persistence of MNV and TV RNA. Bacterial counts were significantly affected by time and plant type but not by the stressors. However, bacterial counts correlated significantly with HuNoV RNA titers on spinach and with the presence of surrogate viruses on both plant types under various conditions. In conclusion, abiotic stressors and phyllosphere bacterial density may differentially influence the survival of HuNoV and its surrogates on lettuce and spinach, emphasizing the need for the use of preventive measures at the preharvest stage.H uman noroviruses (HuNoVs) are the leading cause of acute gastroenteritis and foodborne illnesses in the United States, causing 19 million to 21 million cases, 570 to 800 deaths, and $777 million in health care-associated costs annually (1). These singlestranded RNA viruses belong to the Caliciviridae family and are 28 to 35 nm in diameter and nonenveloped. Although these viruses cause self-limiting gastroenteritis, more serious infections can develop in high-risk groups, such as the elderly and immunocompromised populations (2). The virus is mainly transmitted to humans via the fecal-oral route either by contact with an infected person or fomites or by ingestion of contaminated food and water (2). Leafy greens are frequently associated with HuNoV outbreaks (1) and are globally recognized to be a high priority in terms of the microbial safety of fresh produce (3). Contamination of leafy greens with HuNoVs can occur at any stage along the farm-to-fork chain (3) through a number of sources, including fecally contaminated water used for irrigation, improperly treated sewage sludge used for fertilization, and food harvesters or food handlers who use improper hygiene practices (4, 5). Because HuNoVs have a low infectious dose (ϳ18 to 1,000 viral particles) (6) ...

show abstract

Murine Norovirus: Propagation, Quantification, and Genetic Manipulation

Cited by 86 publications

References 54 publications

Feline Calicivirus, Murine Norovirus, Porcine Sapovirus, and Tulane Virus Survival on Postharvest Lettuce

Feline Calicivirus, Murine Norovirus, Porcine Sapovirus, and Tulane Virus Survival on Postharvest Lettuce

Oral Norovirus Infection Is Blocked in Mice Lacking Peyer's Patches and Mature M Cells

Abiotic Stress and Phyllosphere Bacteria Influence the Survival of Human Norovirus and Its Surrogates on Preharvest Leafy Greens

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