Electron beam inactivation of Tulane virus on fresh produce, and mechanism of inactivation of human norovirus surrogates by electron beam irradiation.

Ionizing radiation, whether by electron beams or gamma rays, is a non-thermal processing technique used to improve the microbial safety and shelf-life of many different food products. This technology is highly effective against bacterial pathogens, but data on its effect against foodborne viruses is limited. A mechanism of viral inactivation has been proposed with gamma irradiation, but no published study discloses a mechanism for electron beam (e-beam). This study had three distinct goals: 1) evaluate the sensitivity of a human norovirus surrogate, Tulane virus (TV), to e-beam irradiation in foods, 2) compare the difference in sensitivity of TV and murine norovirus (MNV-1) to e-beam irradiation, and 3) determine the mechanism of inactivation of these two viruses by e-beam irradiation. TV was reduced from 7 log10 units to undetectable levels at target doses of 16 kGy or higher in two food matrices (strawberries and lettuce). MNV-1 was more resistant to e-beam treatment than TV. At target doses of 4 kGy, e-beam provided a 1.6 and 1.2 log reduction of MNV-1 in phosphate buffered saline (PBS) and Dulbecco's Modified Eagle Medium (DMEM), compared to a 1.5 and 1.8 log reduction of TV in PBS and Opti-MEM, respectively. Transmission electron microscopy revealed that increased e-beam doses negatively affected the structure of both viruses. Analysis of viral proteins by SDS-PAGE found that irradiation also degraded viral proteins. Using RT-PCR, irradiation was shown to degrade viral genomic RNA. This suggests that the mechanism of inactivation of e-beam was likely the same as gamma irradiation as the damage to viral constituents led to inactivation.

Electron-beam inactivation of a norovirus surrogate in fresh produce and model systems.

Norovirus remains the leading cause of foodborne illness, but there is no effective intervention to eliminate viral contaminants in fresh produce. Murine norovirus 1 (MNV-1) was inoculated in either 100 ml of liquid or 100 g of food. The inactivation of MNV-1 by electron-beam (e-beam), or high-energy electrons, at varying doses was measured in model systems (phosphate-buffered saline [PBS], Dulbecco's modified Eagle's medium [DMEM]) or from fresh foods (shredded cabbage, diced strawberries). E-beam was applied at a current of 1.5 mA, with doses of 0, 2, 4, 6, 8, 10, and 12 kGy. The surviving viral titer was determined by plaque assays in RAW 264.7 cells. In PBS and DMEM, e-beam at 0 and 2 kGy provided less than a 1-log reduction of virus. At doses of 4, 6, 8, 10, and 12 kGy, viral inactivation in PBS ranged from 2.37 to 6.40 log, while in DMEM inactivation ranged from 1.40 to 3.59 log. Irradiation of inoculated cabbage showed up to a 1-log reduction at 4 kGy, and less than a 3-log reduction at 12 kGy. On strawberries, less than a 1-log reduction occurred at doses up to 6 kGy, with a maximum reduction of 2.21 log at 12 kGy. These results suggest that a food matrix might provide increased survival for viruses. In foods, noroviruses are difficult to inactivate because of the protective effect of the food matrix, their small sizes, and their highly stable viral capsid.

Recovery of Salmonella spp. from raw produce surfaces using ultrasonication.

Fresh fruits and vegetables have been increasingly associated with outbreaks of foodborne illness. Microorganisms on the surface of raw produce may be difficult to remove for decontamination or microbial sampling due to entrapped or attached cells and porous surfaces. The objective of this study was to determine if ultrasonic treatment using 40 kHz with varying temperatures and agitation times can enhance removal and recovery of Salmonella spp. from raw produce surfaces. Strawberry, apple, and cantaloupe surfaces were spot inoculated with a fivestrain cocktail of nalidixic acid-resistant Salmonella spp. Produce were immersed in 0.1% buffered peptone water in either a Whirl-Pak bag for manual shaking (60 sec) or a sterile beaker for ultrasonic treatment (60 or 120 sec). Diluent temperatures of either 25 degrees C or 40 degrees C were used with these sampling protocols. Diluents were spiral plated onto tryptic soy agar supplemented with 50 ppm of nalidixic acid. No significant differences were observed between each Salmonella recovery method for each produce type. In this study, ultrasonic treatment using 40 kHz did not enhance recovery of Salmonella spp. from produce surfaces. The ultrasonic frequency, temperatures, and times used in this study did not enhance or diminish bacterial recovery. Additional research is still needed to determine the efficacy of other ultrasonic frequencies, diluents, and diluent temperatures for enhancing the recovery and enumeration of Salmonella spp. and other microorganisms from raw produce surfaces.