The current study investigated the effects of heat treatment (85 °C or 100 °C for 5-20 min) on human norovirus (HuNoV) GII.4\'s capsid stability in fresh oysters. In addition, propidium monoazide (PMA) was used in viral samples to distinguish infectious viruses and evaluated using real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR). Further, we explored the effect of the heat treatment on oyster quality (Hunter color and hardness). The titer of HuNoV for oysters significantly (p < 0.05) decreased to 0.39-1.32 and 0.93-2.27 log10 copy number/μL in the non-PMA and PMA-treated groups, respectively, after heat treatment. HuNoV in oysters not treated with PMA showed a decrease of <1.5 - log10, whereas in PMA-treated oysters, a decrease of >1 - log10 was observed after treatment at 85 °C for 10 min. Treatments for both 15 min and 20 min at 100 °C showed a >99% log10 reduction using PMA/RT-qPCR. In the Hunter color, an increase in heat temperature and duration was associated with a significant decrease in \'L\' (brightness+, darkness-) and an increase in \'a\' (redness+, greenness-) and \'b\' (yellowness+, blueness-) (p < 0.05). Our findings confirmed that the hardness of oyster meat significantly increased with increasing temperature and time (p < 0.05). This study demonstrated that PMA/RT-qPCR was effective in distinguishing HuNoV viability in heat-treated oysters. The optimal heat treatment for oysters was 10 min at 85 °C and 5 min at 100 °C.

Human norovirus (HuNoV) is a common cause of outbreaks linked to food. In this study, the effectiveness of a non-thermal method known as high-pressure processing (HPP) on the viable reduction of an HuNoV GII.4 strain on raw crabs was evaluated at three different pressures (200, 400, and 600 MPa). HuNoV viability in raw crabs was investigated by using propidium monoazide/sarkosyl (PMA) as a nucleic acid intercalating dye prior to performing a real-time reverse transcription-polymerase chain reaction (RT-qPCR). The effect of the HPP exposure on pH, sensory, and Hunter colors were also assessed. HuNoV was reduced in raw crabs compared with control to HPP (0.15-1.91 log) in non-PMA and (0.67-2.23 log) in PMA. HuNoV genomic titer reduction was <2 log copy number/µL) when HPP was treated for 5 min without PMA pretreatment, but it was reduced to >2 log copy number/µL after PMA. The pH and Hunter colors of the untreated and HPP-treated raw crabs were significantly different (p < 0.05), but sensory attributes were not significant. The findings indicate that PMA/RT-qPCR could be used to detect HuNoV infectivity without altering the quality of raw crabs after a 5 min treatment with HPP. Therefore, HuNoV GII.4 could be reduced up to 2.23 log in food at a commercially acceptable pressure duration of 600 MPa for 5 min.

Human norovirus (HNoV) GII.4 and Vibrio parahaemolyticus may be found in sea squirts. Antimicrobial effects of floating electrode-dielectric barrier discharge (FE-DBD) plasma (5-75 min, N2 1.5 m/s, 1.1 kV, 43 kHz) treatment were examined. HNoV GII.4 decreased by 0.11-1.29 log copy/μL with increasing duration of treatment time, and further by 0.34 log copy/μL when propidium monoazide (PMA) treatment was added to distinguish infectious viruses. The decimal reduction time (D1) of non-PMA and PMA-treated HNoV GII.4 by first-order kinetics were 61.7 (R2 = 0.97) and 58.8 (R2 = 0.92) min, respectively. V. parahaemolyticus decreased by 0.16-1.5 log CFU/g as treatment duration increased. The D1 for V. parahaemolyticus by first-order kinetics was 65.36 (R2 = 0.90) min. Volatile basic nitrogen showed no significant difference from the control until 15 min of FE-DBD plasma treatment, increasing after 30 min. The pH did not differ significantly from the control by 45-60 min, and Hunter color in \"L\" (lightness), \"a\" (redness), and \"b\" (yellowness) values reduced significantly as treatment duration increased. Textures appeared to be individual differences but were not changed by treatment. Therefore, this study suggests that FE-DBD plasma has the potential to serve as a new antimicrobial to foster safer consumption of raw sea squirts.

This study investigated the antiviral effects of floating electrode-dielectric barrier discharge (FE-DBD) plasma treatment (1.1 kV, 43 kHz, N2 1.5 m/s, 5-30 min) against human norovirus (HuNoV) GII.4 in Jogaejeotgal Infectivity was assessed using real-time quantitative-PCR (RT-qPCR) following treatment of samples with propidium monoazide (PMA) and sodium lauroyl sarcosinate (Sarkosyl). This study also investigated the effects of FE-DBD plasma treatment on Jogaejeotgal quality (assessed using pH value and Hunter colors). Following inoculation, the average titers of HuNoV GII.4 in Jogaejeotgal significantly (P < 0.05) decreased with increases in the FE-DBD plasma treatment time in both the non-PMA-treated and PMA + Sarkosyl-treated samples; in the non-PMA and PMA + Sarkosyl treated Jogaejeotgal, HuNoV GII.4 titers (log10 copy number/µL) were to: 3.16 and 2.95 (5 min), 2.90 and 2.48 (10 min), 2.82 and 2.40 (15 min), 2.58 and 2.26 (20 min), 2.48 and 2.06 (25 min), and 2.23 and 1.91 (30 min), respectively. The average titers of HuNoV demonstrated significant (P < 0.05) reductions of 0.35 log10 (55.3%) in PMA + Sarkosyl-treated samples compared with the non-PMA treated samples following exposure to 5-30 min of FE-DBD plasma. Reductions of >1-log for HuNoV in PMA + Sarkosyl- treated Jogaejeotgal required treatments of FE-DBD of 5-30 min. Using the first order kinetic model (R2 = 0.95), GII.4 decimal reduction time (D-value) resulting from FE-DBD plasma was 23.75 min. The pH and Hunter colors (\"L\", \"a\", and \"b\") were not significantly different (P > 0.05) between the untreated and FE-DBD plasma-treated Jogaejeotgal. Based on these results, the PMA + Sarkosyl/RT-qPCR method could be assessing HuNoV viability following 5-30 min treatment of FE-DBD plasma. Furthermore, may be an optimal treatment for Jogaejeotgal without altering the food quality (color and pH).