The effect of superchilled storage at -1°C on the microbial safety of oyster depurated with 0.2, 0.4, and 0.6 mg/L ozone was studied for 14 days. Fecal coliforms (4,100–16,000 MPN/100 g), Escherichia coli (1,500–3,650 MPN/100 g), Vibrio cholerae non-O1/non-O139 (13.0–102.0 MPN/g), and Salmonella spp. (2.270–3.035 × 103 CFU/g) were initially present in raw oysters. After 6 h depuration, fecal coliform counts decreased (P < 0.05) to 300, 20 and 20 MPN/100 g for 0.2, 0.4, and 0.6 mg/L treatments, while a 0.3 log decrease in control oysters was observed. Initial E. coli counts decreased (P < 0.05) in oysters to 50, 20, and 20 MPN/100 g for 0.2, 0.4, and 0.6 mg/L treatments, respectively. A 1 log reduction in V. cholerae non-O1/non-139 levels were observed in 0.4 and 0.6 mg/L-treatments after 2 and 4 h depuration. Salmonella spp. was not detected in oyster samples after 6 h depuration in 0.4 and 0.6 mg/L-ozone treatments. Considering the bacterial loads after depuration, at the end of superchilled storage the 0.4 mg/L-ozonated oysters attained lower (P < 0.05) fecal coliform levels (280 MPN/100 g) and E. coli counts in 0.4 and 0.6 mg/L-ozonated oysters (20 and 95 MPN/100 g, respectively). A 2-log decrease in V. cholerae non-O1/non-O139 levels on day 5 in 0.4 and 0.6 mg/L-ozonated oysters (< 0.3 MPN/g) was attained. V. cholerae non-O1/non-O139 counts in control oysters decreased 1 log on day 9 of superchilled storage. Salmonella spp. was not detected in ozonated and superchilled stored oysters. Levels of fecal coliforms, E. coli, Salmonella spp., and V. cholerae non-O1/non-O139 in non-ozone depurated oyster samples were higher than in control, 0.4 and 0.6 mg/L ozonated oyster samples during superchilled storage. The cumulative mortality rates after 14 days of storage for superchilled oysters (22.2%) was higher (P < 0.05) than 0.6 mg/L O3 (7.2%) and 0.4 mg/L O3 (5.8%) treatments, and control oysters (5.6%). pH values in control oysters decreased significantly (P < 0.05) throughout the storage period but not in oysters of both ozone treatments, indicating no detrimental effects on oyster survival. The results of this study suggest that superchilled storage enables ozonated shellstock oysters (0.4 mg/L-6 h) stored for 9 days to be safe human consumption.

Expression of the regulatory stress rpoS gene controls the transcription of cspA genes, which are involved in survival and adaptation to low temperatures. The purpose of this study was to assess the growth kinetics of naturally occurring V. parahaemolyticus in shellstock oysters and in vitro and the cold-shock-induced expression of the rpoS and cspA gene response in vitro during postharvest refrigeration. Naturally contaminated eastern oysters (Crassostrea virginica) and pathogenic (Vp-tdh) and nonpathogenic (Vp-tlh) isolates were stored at 7 ± 1 °C for 168 h and 216 h, respectively. The regulatory stress (rpos) and cold-shock (cspA) gene expressions were determined by reverse transcription PCR. At 24 h, the (Vp-tdh) strain grew faster (p < 0.05) than the (Vp-tlh) strain in oysters (λ = 0.33, 0.39, respectively) and in vitro (λ = 0.89, 37.65, respectively), indicating a better adaptation to cold shock for the (Vp-tdh) strain in live oysters and in vitro. At 24 h, the (Vp-tdh) strain rpoS and cspA gene expressions were upregulated by 1.9 and 2.3-fold, respectively, but the (Vp-tlh) strain rpoS and cspA gene expressions were repressed and upregulated by −0.024 and 1.9-fold, respectively. The V. parahaemolyticus strains that were isolated from tropical oysters have adaptive expression changes to survive and grow at 7 °C, according to their virulence.