Physicochemical Parameters Affecting Norovirus Adhesion to Ready-To-Eat Foods.

The adhesion of noroviruses to strawberry, turkey slices, ham, and cheddar cheese was studied using murine norovirus 1 (MNV-1) as a surrogate for human norovirus (NoV). Based on plaque assay, the recovery and adhesion of MNV-1 depended on the food type (turkey versus strawberry), pH of the initial suspension buffer (pH 4 versus pH 7), and food fat composition (C8 versus C18). Recovery of infectious particles from turkey was 68% compared to 9.4% from strawberry. On turkey, adhesion of MNV-1 was lower at pH 7 (pH of fecal matter), and virus particles adhered to this pH were recovered more easily (33,875 PFU) than at pH 4 (pH of vomitus). The presence of fat and the composition of fatty acids seemed to increase MNV-1 recovery and adherent viral particles recovered but did not affect adhesion (68% on fat-free turkey and regular turkey). Adherent MNV-1 particles recovered from stainless steel coated with saturated fatty acid (C8, C14, C18) increased significantly with chain length (P < 0.05), but adhesion did not seem to change. Using liquid droplet contact angle to measure surface energy, it was deduced that hydrophobic interactions contribute considerably to the adhesion of MNV-1 to stainless steel, polyvinyl chloride, and high-density polyethylene. IMPORTANCE Ready-to-eat (RTE) foods are major vehicles of transmission of foodborne viral pathogens, including NoV. The high incidence of gastroenteritis caused by viruses is due largely to their persistence in the environment and adhesion to different kinds of surfaces in the food industry, including the foods themselves. Compared with bacteria, adhesion of viruses to surfaces is poorly understood. Better knowledge of the physicochemical parameters involved in the adhesion of NoV to ready-to-eat foods is essential to devising effective strategies for reducing the persistence and, thus, the transmission of this virus.

Inactivation of Murine Norovirus Suspended in Organic Matter Simulating Actual Conditions of Viral Contamination.

Foodborne viral illnesses are frequent worldwide and costly for the society. Human norovirus is one of the most common causal agents. Although some norovirus genotypes can now be cultured, surrogates are still used for inactivation studies. The aim of this study was to evaluate the effects of different organic loads individually (artificial feces, real fecal matter, ASTM tripartite organic load, fetal bovine serum) on the efficacy of three highly used sanitization treatments (thermal inactivation, peracetic acid and sodium hypochlorite treatment) using murine norovirus 3 in solutions and surfaces. Based on plaque-forming units, we show that organic matter protects murine norovirus 3 against thermal inactivation (viral reduction of ~ 1 log compared to 2.67 with PBS). However, there was a low-level but significant protection against peracetic acid (viral reduction of ~ 2 log compared to 2.85 with PBS) and none in the presence of sodium hypochlorite. Our study showed that the tested organic matters do not behave similarly depending on the treatments, especially with heat treatments, which showed a higher protection. Furthermore, Feclone ™ artificial feces mimicked some aspect of real fecal matter and may be used instead. Our results will be helpful to researchers undertaking viral inactivation studies in which an organic matrix is used to simulate actual conditions of human norovirus environment.

Efficacy of oxidizing disinfectants at inactivating murine norovirus on ready-to-eat foods.

Noroviruses are the leading cause of foodborne illness, and ready-to-eat foods are frequent vehicles of their transmission. Studies of the disinfection of fruits and vegetables are becoming numerous. It has been shown that strong oxidizing agents are more effective than other chemical disinfectants for inactivating enteric viruses. The aim of this study was to evaluate the efficacy of oxidizing disinfectants (sodium hypochlorite, chloride dioxide and peracetic acid) at inactivating noroviruses on fruits and vegetables, using a norovirus surrogate, namely murine norovirus 3, which replicates in cell culture. Based on plaque assay, solutions of peracetic acid (85 ppm) and chlorine dioxide (20 ppm) reduced the infectivity of the virus in suspension by at least 3 log10 units after 1 min, while sodium hypochlorite at 50 ppm produced a 2-log reduction. On the surface of blueberries, strawberries and lettuce, chlorine dioxide was less effective than peracetic acid and sodium hypochlorite, which reduced viral titers by approximately 4 logs. A surprising increase in the efficacy of sodium hypochlorite on surfaces fouled with artificial feces was noted.

Comparison of RNA extraction methods for the detection of a norovirus surrogate in ready-to-eat foods.

Four nucleic acid extraction methods were evaluated for the purpose of quantifying a norovirus surrogate (murine norovirus [MNV-1]) concentrated from different food samples. Simple (strawberries and lettuce) and complex (sliced turkey breast, soft-shell clams, and potato salad) food matrices were inoculated with a viral suspension containing high (4×10(5) PFU) or low (4×10(3) PFU) numbers of viral particles. MNV-1 was eluted using either the Pulsifier™ or repetitive pipetting. The four methods were based on using magnetic silica (MiniMAG), non-magnetic silica (bioMérieux Basic kit), silica membrane (Qiagen kit), and phenol (TriReagent) for RNA extraction. The greatest recovery of viral RNA from simple matrices was obtained using magnetic silica for both inoculation levels. For strawberries, the addition of pectinase during the elution step improved RNA recovery when the Pulsifier was used with silica membrane extraction and when repetitive pipetting was used with magnetic silica extraction. In the case of complex matrices, the extraction of high or low numbers of MNV-1 was highest overall using magnetic silica. The exception was soft-shell clams with a high viral load, in which the greatest recovery was obtained with the phenol-based method. In general, magnetic silica was the most effective for extracting both high and low numbers of MNV-1 particles from a wide range of foods.

Attachment of noroviruses to stainless steel and their inactivation, using household disinfectants.

The aims of this study were (i) to evaluate the impact of pH and relative humidity on the attachment of norovirus (NoV) to fomites and (ii) to evaluate the effectiveness of different household disinfectants on NoV attached to fomites. Plaque assay and/or real-time reverse transcription PCR assay were used to determine the amount of murine and human NoV attached to stainless steel disks, i.e., the amount removed by sonication in elution buffer but not by surface rinses with water only. An enzymatic pretreatment was used for both human and murine NoV before the real-time reverse transcription PCR assay to avoid detection of RNA associated with inactivated virus. For both murine and human NoV, maximum attachment was obtained after a contact time of 10 min. Attachment of NoV to stainless steel does not appear to be affected by pH, although murine NoV was less attached (<2 log units) at pH 9 and at low relative humidity (25%) than was human NoV (3 log units). Sodium hypochlorite (3%) was the most effective disinfectant, producing a greater than 3-log reduction after 10 min compared with less than a 1-log reduction after treatment with quaternary ammonium compounds and ethoxylated alcohols. Murine NoV was more sensitive than human NoV to disinfectants by approximately 1 to 2 log units. These results will help improve strategies for decontaminating surfaces harboring NoV and thus reduce the incidence of illness caused by these pathogens in the food sector and domestic environments.