Effect of weather on the die-off of Escherichia coli and attenuated Salmonella Typhimurium on preharvest leafy greens following irrigation with contaminated water.

The Food Safety Modernization Act (FSMA) includes a time-to-harvest interval following the application of non-compliant water to pre-harvest produce to allow for microbial die-off. However, additional scientific evidence is needed to support this rule. This study aimed to determine the impact of weather on the die-off rate of E. coli and Salmonella on spinach and lettuce under field conditions. Standardized, replicated field trials were conducted in California, New York, and Spain over two years. Baby spinach and lettuce were grown and inoculated with a ∼104 CFU/mL cocktail of E. coli and attenuated Salmonella Leaf samples were collected at 7 timepoints (0-96h) following inoculation; E. coli and Salmonella were enumerated. The associations of die-off with study design factors (location, produce type, and bacteria) and weather were assessed using log-linear and biphasic segmented log-linear regression. A segmented log-linear model best fit die-off on inoculated leaves in most cases, with a greater variation in the segment 1 die-off rate across trials [-0.46 (95% confidence interval (95% CI): -0.52, -0.41) to -6.99 (95% CI: -7.38, -6.59) log10 die-off/day] compared to the segment 2 die-off rate [0.28 (95% CI: -0.20, 0.77) to -1.00 (95% CI: -1.16, -0.85) log10 die-off/day]. A lower relative humidity was associated with a faster segment 1 die-off and an earlier breakpoint (the time when segment 1 die-off rate switches to the segment 2 rate). Relative humidity was also found to be associated with whether die-off would comply with FSMA's specified die-off rate of -0.5 log10 die-off/day.Importance The log-linear die-off rate proposed by FSMA is not always appropriate, as the die-off of foodborne bacterial pathogens and specified agricultural water quality indicator organisms appear to commonly follow a biphasic die-off pattern with an initial rapid decline followed by a period of tailing. While we observed substantial variation in the net culturable population levels of Salmonella and E. coli at each time point, die-off rate and FSMA compliance (i.e., at least a 2 log10 die-off over 4 days) appear to be impacted by produce type, bacteria, and weather; die-off on lettuce tended to be faster than that on spinach, die-off of E. coli tended to be faster than that of attenuated Salmonella, and die-off tended to become faster as relative humidity decreased. As such, the use of a single die-off rate for estimating time-to-harvest intervals across different weather conditions, produce types, and bacteria should be revised.