Surrogates for the Study of Norovirus Stability and Inactivation in the Environment: A Comparison of Murine Norovirus and Feline Calicivirus

2006 ◽  
Vol 69 (11) ◽  
pp. 2761-2765 ◽  
Author(s):  
JENNIFER L. CANNON ◽  
EFSTATHIA PAPAFRAGKOU ◽  
GEUNWOO W. PARK ◽  
JASON OSBORNE ◽  
LEE-ANN JAYKUS ◽  
...  
Keyword(s):  
Cell Culture ◽  
Thermal Inactivation ◽  
Genetic Relatedness ◽  
Mammalian Cell Culture ◽  
Feline Calicivirus ◽  
Murine Norovirus ◽  
Log Reduction ◽  
Culture Model ◽  
Dry Conditions ◽  
Ph Range

Human noroviruses (NoVs) are the leading cause of food- and waterborne outbreaks of acute nonbacterial gastroenteritis worldwide. As a result of the lack of a mammalian cell culture model for these viruses, studies on persistence, inactivation, and transmission have been limited to cultivable viruses, including feline calicivirus (FCV). Recently, reports of the successful cell culture of murine norovirus 1 (MNV-1) have provided investigators with an alternative surrogate for human NoVs. In this study, we compared the inactivation profiles of MNV-1 to FCV in an effort to establish the relevance of MNV-1 as a surrogate virus. Specifically, we evaluated (i) stability upon exposure to pH extremes; (ii) stability upon exposure to organic solvents; (iii) thermal inactivation; and (iv) surface persistence under wet and dry conditions. MNV-1 was stable across the entire pH range tested (pH 2 to 10) with less than 1 log reduction in infectivity at pH 2, whereas FCV was inactivated rapidly at pH values <3 and >9. FCV was more stable than MNV-1 at 56°C, but both viruses exhibited similar inactivation at 63 and 72°C. Long-term persistence of both viruses suspended in a fecal matrix and inoculated onto stainless steel coupons were similar at 4°C, but at room temperature in solution, MNV-1 was more stable than FCV. The genetic relatedness of MNV-1 to human NoVs combined with its ability to survive under gastric pH levels makes this virus a promising and relevant surrogate for studying environmental survival of human NoVs.

Hydrogen Peroxide Vapor Decontamination in a Patient Room Using Feline Calicivirus and Murine Norovirus as Surrogate Markers for Human Norovirus

2016 ◽  
Vol 37 (5) ◽  
pp. 561-566 ◽  
Author(s):  
Torsten Holmdahl ◽  
Mats Walder ◽  
Nathalie Uzcátegui ◽  
Inga Odenholt ◽  
Peter Lanbeck ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Cell Culture ◽  
Tissue Culture ◽  
Feline Calicivirus ◽  
Murine Norovirus ◽  
Patient Room ◽  
Human Norovirus ◽  
Log Reduction ◽  
Infective Dose ◽  
Hydrogen Peroxide Vapor

OBJECTIVETo determine whether hydrogen peroxide vapor (HPV) could be used to decontaminate caliciviruses from surfaces in a patient room.DESIGNFeline calicivirus (FCV) and murine norovirus (MNV) were used as surrogate viability markers to mimic the noncultivable human norovirus. Cell culture supernatants of FCV and MNV were dried in triplicate 35-mm wells of 6-well plastic plates. These plates were placed in various positions in a nonoccupied patient room that was subsequently exposed to HPV. Control plates were positioned in a similar room but were never exposed to HPV.METHODSVirucidal activity was measured in cell culture by reduction in 50% tissue culture infective dose titer for FCV and by both 50% tissue culture infective dose titer and plaque reduction for MNV.RESULTSNeither viable FCV nor viable MNV could be detected in the test room after HPV treatment. At least 3.65 log reduction for FCV and at least 3.67 log reduction for MNV were found by 50% tissue culture infective dose. With plaque assay, measurable reduction for MNV was at least 2.85 log units.CONCLUSIONSThe successful inactivation of both surrogate viruses indicates that HPV could be a useful tool for surface decontamination of a patient room contaminated by norovirus. Hence nosocomial spread to subsequent patients can be avoided.Infect Control Hosp Epidemiol 2016;37:561–566

scholarly journals Evaluation of Murine Norovirus, Feline Calicivirus, Poliovirus, and MS2 as Surrogates for Human Norovirus in a Model of Viral Persistence in Surface Water and Groundwater

2007 ◽  
Vol 74 (2) ◽  
pp. 477-484 ◽  
Author(s):  
Jinhee Bae ◽  
Kellogg J. Schwab
Keyword(s):  
Cell Culture ◽  
Surface Water ◽  
Multiple Linear Regression Model ◽  
Viral Persistence ◽  
Environmental Stability ◽  
Great Promise ◽  
Feline Calicivirus ◽  
Virus Infectivity ◽  
Murine Norovirus ◽  
Surface Water And Groundwater

ABSTRACT Human noroviruses (NoVs) are a significant cause of nonbacterial gastroenteritis worldwide, with contaminated drinking water a potential transmission route. The absence of a cell culture infectivity model for NoV necessitates the use of molecular methods and/or viral surrogate models amenable to cell culture to predict NoV inactivation. The NoV surrogates murine NoV (MNV), feline calicivirus (FCV), poliovirus (PV), and male-specific coliphage MS2, in conjunction with Norwalk virus (NV), were spiked into surface water samples (n = 9) and groundwater samples (n = 6). Viral persistence was monitored at 25°C and 4°C by periodically analyzing virus infectivity (for all surrogate viruses) and nucleic acid (NA) for all tested viruses. FCV infectivity reduction rates were significantly higher than those of the other surrogate viruses. Infectivity reduction rates were significantly higher than NA reduction rates at 25°C (0.18 and 0.09 log10/day for FCV, 0.13 and 0.10 log10/day for PV, 0.12 and 0.06 log10/day for MS2, and 0.09 and 0.05 log10/day for MNV) but not significant at 4°C. According to a multiple linear regression model, the NV NA reduction rates (0.04 ± 0.01 log10/day) were not significantly different from the NA reduction rates of MS2 (0.05 ± 0.03 log10/day) and MNV (0.04 ± 0.03 log10/day) and were significantly different from those of FCV (0.08 ± 0.03 log10/day) and PV (0.09 ± 0.03 log10/day) at 25°C. In conclusion, MNV shows great promise as a human NoV surrogate due to its genetic similarity and environmental stability. FCV was much less stable and thus questionable as an adequate surrogate for human NoVs in surface water and groundwater.

scholarly journals Quantitative form and fit of N95 filtering facepiece respirators are retained and coronavirus surrogate is inactivated after heat treatments

2020 ◽  
Author(s):  
Travis Massey ◽  
Monica Borucki ◽  
Samuel Paik ◽  
Kyle Fuhrer ◽  
Mihail Bora ◽  
...  
Keyword(s):  
Thermal Inactivation ◽  
Hepatitis Virus ◽  
Filter Material ◽  
Healthcare Facilities ◽  
Emergency Situations ◽  
Log Reduction ◽  
Quantitative Form ◽  
Dry Conditions ◽  
Dry Heating ◽  
Single Use

SummaryRe-use of filtering facepiece respirators (FFRs, commonly referred to as N95s) normally meant for single use only is becoming common in healthcare facilities due to shortages caused by the COVID19 pandemic. Here we report that mouse hepatitis virus (MHV) initially seeded on FFR filter material is inactivated (6 log reduction as measured by 50% tissue culture infective dose (TCID50)) after dry heating at 75 ºC for 30 minutes. We also find that the quantitative fit of FFRs after heat treatment at this temperature, under dry conditions or at 90% relative humidity, is not affected by single or ten heating cycles. Previous studies have reported that the filtration efficiency of FFR filters is not negatively impacted by these heating conditions. These results suggest that thermal inactivation of coronaviruses is a potentially rapid and widely deployable method to re-use N95 FFRs in emergency situations where re-using FFRs is a necessity and broad-spectrum sterilization is unavailable. However, we also found that a heat source that emits radiation (e.g., an exposed heating element) results in rapid qualitative degradation of the FFR. Finally, we discuss differences in the results reported here and other recent studies investing heat as a means to recycle FFRs and suggest that overall wear time and donning/doffing cycles are important factors that need to be considered.

Recovery and Disinfection of Two Human Norovirus Surrogates, Feline Calicivirus and Murine Norovirus, from Hard Nonporous and Soft Porous Surfaces

2015 ◽  
Vol 78 (10) ◽  
pp. 1842-1850 ◽  
Author(s):  
THOMAS YEARGIN ◽  
ANGELA FRASER ◽  
GUOHUI HUANG ◽  
XIUPING JIANG
Keyword(s):  
Sodium Hypochlorite ◽  
Limit Of Detection ◽  
Plaque Assay ◽  
Viral Rna ◽  
Feline Calicivirus ◽  
Murine Norovirus ◽  
Human Norovirus ◽  
Surface Type ◽  
Log Reduction ◽  
Porous Surfaces

Human norovirus is a leading cause of foodborne disease and can be transmitted through many routes, including environmental exposure to fomites. In this study, both the recovery and inactivation of two human norovirus surrogates, feline calicivirus (FCV) and murine norovirus (MNV), on hard nonporous surfaces (glass) and soft porous surfaces (polyester and cotton) were evaluated by both plaque assay and reverse transcription quantitative PCR method. Two disinfectants, sodium hypochlorite (8.25%) and accelerated hydrogen peroxide (AHP, at 4.25%) were evaluated for disinfection efficacy. Five coupons per surface type were used to evaluate the recovery of FCV and MNV by sonication and stomaching and the disinfection of each surface type by using 5 ml of disinfectant for a contact time of 5 min. FCV at an initial titer of ca. 7 log PFU/ml was recovered from glass, cotton, and polyester at 6.2, 5.4, and 3.8 log PFU/ml, respectively, compared with 5.5, 5.2, and 4.1 log PFU/ml, respectively, for MNV with an initial titer of ca. 6 log PFU/ml. The use of sodium hypochlorite (5,000 ppm) was able to inactivate both FCV and MNV (3.1 to 5.5 log PFU/ml) below the limit of detection on all three surface types. AHP (2,656 ppm) inactivated FCV (3.1 to 5.5 log PFU/ml) below the limit of detection for all three surface types but achieved minimal inactivation of MNV (0.17 to 1.37 log PFU/ml). Reduction of viral RNA by sodium hypochlorite corresponded to 2.72 to 4.06 log reduction for FCV and 2.07 to 3.04 log reduction for MNV on all three surface types. Reduction of viral RNA by AHP corresponded to 1.89 to 3.4 log reduction for FCV and 0.54 to 0.85 log reduction for MNV. Our results clearly indicate that both virus and surface types significantly influence recovery efficiency and disinfection efficacy. Based on the performance of our proposed testing method, an improvement in virus recovery will be needed to effectively validate virus disinfection of soft porous surfaces.

Determination of the Thermal Inactivation Kinetics of the Human Norovirus Surrogates, Murine Norovirus and Feline Calicivirus

2013 ◽  
Vol 76 (1) ◽  
pp. 79-84 ◽  
Author(s):  
HAYRIYE BOZKURT ◽  
DORIS H. D'SOUZA ◽  
P. MICHAEL DAVIDSON
Keyword(s):  
Thermal Inactivation ◽  
Weibull Model ◽  
Inactivation Kinetics ◽  
Feline Calicivirus ◽  
Murine Norovirus ◽  
Order Model ◽  
Human Norovirus ◽  
First Order ◽  
D Values ◽  
Kinetics Of

Studies are needed to bridge existing data gaps and determine appropriate parameters for thermal inactivation methods for human noroviruses. Cultivable surrogates, such as feline calicivirus (FCV-F9) and murine norovirus (MNV-1), have been used in the absence of human norovirus infectivity assays. This study aimed to characterize the thermal inactivation kinetics of MNV-1 and FCV-F9 at 50, 56, 60, 65, and 72°C for different treatment times (0 to 60 min). Thermal inactivation was performed using the capillary tube method with titers of 4.0 × 107 (MNV-1) and 5.8 × 108 (FCV-F9) PFU/ml in triplicate experiments, followed by standard plaque assays in duplicate for each experiment. Weibull and first-order models were compared to describe survival curve kinetics. Model fitness was investigated by comparing the regression coefficients (R2) and the chi-square (χ2) and root mean square error (RMSE) values. The D-values calculated from the first-order model (50 to 72°C) were 0.15 to 34.49 min for MNV-1 and 0.11 to 20.23 min for FCV-9. Using the Weibull model, the tD values needed to destroy 1 log PFU of MNV-1 and FCV-F9 at the same temperatures were 0.11 to 28.26 and 0.06 to 13.86 min, respectively. In terms of thermal resistance, MNV-1 was more sensitive than FCV-F9 up to 65°C. At 72°C, FCV-F9 was slightly more susceptible to heat inactivation. Results revealed that the Weibull model was more appropriate to represent the thermal inactivation behavior of both tested surrogates. The z-values were calculated using D-values for the first-order model and the tD values for the Weibull model. The z-values were 9.31 and 9.19°C for MNV-1 and 9.36 and 9.31°C for FCV-F9 for the first-order and Weibull models, respectively. This study provides more precise information than previous reports on the thermal inactivation kinetics of two norovirus surrogates for use in thermal process calculations.

scholarly journals Protein-RNA linkage and posttranslational modifications of feline calicivirus and murine norovirus VPg proteins

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e2134 ◽  
Author(s):  
Allan Olspert ◽  
Myra Hosmillo ◽  
Yasmin Chaudhry ◽  
Lauri Peil ◽  
Erkki Truve ◽  
...  
Keyword(s):  
Cell Culture ◽  
Life Cycle ◽  
Serine Phosphorylation ◽  
Viral Rna ◽  
Feline Calicivirus ◽  
Murine Norovirus ◽  
Wide Range ◽  
Positive Sense ◽  
Covalently Linked

Members of theCaliciviridaefamily of positive sense RNA viruses cause a wide range of diseases in both humans and animals. The detailed characterization of the calicivirus life cycle had been hampered due to the lack of robust cell culture systems and experimental tools for many of the members of the family. However, a number of caliciviruses replicate efficiently in cell culture and have robust reverse genetics systems available, most notably feline calicivirus (FCV) and murine norovirus (MNV). These are therefore widely used as representative members with which to examine the mechanistic details of calicivirus genome translation and replication. The replication of the calicivirus RNA genome occurs via a double-stranded RNA intermediate that is then used as a template for the production of new positive sense viral RNA, which is covalently linked to the virus-encoded protein VPg. The covalent linkage to VPg occurs during genome replication via the nucleotidylylation activity of the viral RNA-dependent RNA polymerase. Using FCV and MNV, we used mass spectrometry-based approach to identify the specific amino acid linked to the 5′ end of the viral nucleic acid. We observed that both VPg proteins are covalently linked to guanosine diphosphate (GDP) moieties via tyrosine positions 24 and 26 for FCV and MNV respectively. These data fit with previous observations indicating that mutations introduced into these specific amino acids are deleterious for viral replication and fail to produce infectious virus. In addition, we also detected serine phosphorylation sites within the FCV VPg protein with positions 80 and 107 found consistently phosphorylated on VPg-linked viral RNA isolated from infected cells. This work provides the first direct experimental characterization of the linkage of infectious calicivirus viral RNA to the VPg protein and highlights that post-translational modifications of VPg may also occur during the viral life cycle.

scholarly journals Protein-RNA linkage and posttranslational modifications of feline calicivirus and murine norovirus VPg proteins

2016 ◽  
Author(s):  
Allan Olspert ◽  
Myra Hosmillo ◽  
Yasmin Chaudhry ◽  
Lauri Peil ◽  
Erkki Truve ◽  
...  
Keyword(s):  
Cell Culture ◽  
Life Cycle ◽  
Serine Phosphorylation ◽  
Viral Rna ◽  
Feline Calicivirus ◽  
Genome Replication ◽  
Murine Norovirus ◽  
Wide Range ◽  
Positive Sense ◽  
Covalently Linked

Members of the Caliciviridae family of positive sense RNA viruses cause a wide range of diseases in both humans and animals. The detailed characterization of the calicivirus life cycle had been hampered due to the lack of robust cell culture systems and experimental tools for many of the members of the family. However a number of caliciviruses replicate efficiently in cell culture and have robust reverse genetics systems available, most notable feline calicivirus (FCV) and murine norovirus (MNV). These are therefore widely used as representative members with which to examine the mechanistic details of calicivirus genome translation and replication. The replication of the calicivirus RNA genome occurs via a double stranded RNA intermediate in the cytoplasm of the infected cell which is then used as a template for the production of new positive sense viral RNA, which is covalently linked to the virus-encoded protein VPg. The covalent linkage to VPg occurs during genome replication via the nucleotidylylation activity of the viral RNA-dependent RNA polymerase. Using FCV and MNV, we used mass spectrometry-based approach to identify the specific amino acid linked to the 5’ end of the viral nucleic acid. We observed that both VPg proteins are covalently linked to GDP moieties via tyrosine positions 24 and 26 for FCV and MNV respectively. These data fit with previous observations indicating that mutations introduced into these specific amino acids are deleterious for viral replication and fail to produce infectious virus. In addition, we also detected serine phosphorylation sites within the FCV VPg protein with positions 80 and 107 found consistently phosphorylated on VPg-linked viral RNA isolated from infected cells. This work provides the first direct experimental characterisation of the linkage of infectious calicivirus viral RNA to the VPg protein and highlights that post-translational modifications of VPg may also occur during the viral life cycle.

scholarly journals Hsp110 mitigates α-synuclein pathology in vivo

2019 ◽  
Vol 116 (48) ◽  
pp. 24310-24316 ◽  
Author(s):  
Yumiko V. Taguchi ◽  
Erica L. Gorenberg ◽  
Maria Nagy ◽  
Drake Thrasher ◽  
Wayne A. Fenton ◽  
...  
Keyword(s):  
Cell Culture ◽  
Molecular Chaperones ◽  
Mammalian Cell Culture ◽  
Lewy Bodies ◽  
Cell Culture Model ◽  
Culture Model ◽  
Presynaptic Protein

Parkinson’s disease is characterized by the aggregation of the presynaptic protein α-synuclein and its deposition into pathologic Lewy bodies. While extensive research has been carried out on mediators of α-synuclein aggregation, molecular facilitators of α-synuclein disaggregation are still generally unknown. We investigated the role of molecular chaperones in both preventing and disaggregating α-synuclein oligomers and fibrils, with a focus on the mammalian disaggregase complex. Here, we show that overexpression of the chaperone Hsp110 is sufficient to reduce α-synuclein aggregation in a mammalian cell culture model. Additionally, we demonstrate that Hsp110 effectively mitigates α-synuclein pathology in vivo through the characterization of transgenic Hsp110 and double-transgenic α-synuclein/Hsp110 mouse models. Unbiased analysis of the synaptic proteome of these mice revealed that overexpression of Hsp110 can override the protein changes driven by the α-synuclein transgene. Furthermore, overexpression of Hsp110 is sufficient to prevent endogenous α-synuclein templating and spread following injection of aggregated α-synuclein seeds into brain, supporting a role for Hsp110 in the prevention and/or disaggregation of α-synuclein pathology.

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