Human noroviruses (HNoVs) cause significant gastrointestinal disease outbreaks worldwide. Tulane virus (TV) is a cultivable HNoV surrogate widely used to determine control measures against HNoVs. The objective of this study was to determine the heat inactivation kinetics (D‐ and z‐values) of TV in cell‐culture media and on spiked homogenized spinach using the first‐order and Weibull models. TV in cell‐culture media at approximately 7 log PFU/mL (PFU—plaque forming unit) in 2‐mL glass vials was heated at 52, 54, and 56 °C for up to 10 min in a circulating water bath. Survivors were enumerated using confluent host LLC‐MK2 cells in six‐well plates by plaque assay. Data from three replicate treatments assayed in duplicate were analyzed statistically. D‐values by the first‐order model for TV in cell‐culture media at 52, 54, and 56 °C were 4.59 ± 0.05, 2.91 ± 0.05, and 1.74 ± 0.07 min, respectively, with a z‐value of 9.09 ± 0.01 °C (R2 = 0.997). The Weibull model showed td = 1 values of 2.53 ± 0.08, 1.99 ± 0.10, and 0.57 ± 0.64 min, respectively, at the same temperatures. The D‐values for TV in spinach were 7.94 ± 0.21, 4.09 ± 0.04, and 1.43 ± 0.02 min and the z‐value was 10.74 ± 0.01 °C (R2 = 0.98) by the first‐order model and 4.89 ± 0.02, 3.21 ± 0.45, and 0.25 ± 0.38 min for the Weibull model at 50, 54, and 58 °C, respectively. In comparison to previously reported results for the cultivable HNoV surrogate, murine norovirus ‐1, TV in cell‐culture media and spiked on spinach homogenates showed lower D‐ and z‐values. TV may not be an ideal HNoV surrogate for heat inactivation studies in cell‐culture media or homogenized spinach in vacuum bags.
Aichi virus (AiV) that results in gastroenteritis worldwide, is spread through contaminated shellfish and water. The resistance/tolerance of AiV to common inactivation processes along with the absence of commercially available vaccines makes it necessary to study its thermal inactivation kinetics. This research evaluated the heat inactivation of AiV in cell-culture media using 2-ml sterile glass vials by the linear and Weibull models. Heat treatments of AiV titers of 7 log plaque forming units (PFU)/ml were conducted thrice in a water-bath at 50, 54, and 58 • C for up to 90 min. Plaque assays for each dilution in duplicate were used to determine infectious virus titers. Linear model D-values for AiV at 50 ± 1 • C (± = standard error) (come-up time = 68 s), 54 ± 0.7 • C (130 s), and 58 ± 0.6 • C (251 s) were 43.3 ± 4.23 (R 2 = 0.40, RMSE = 0.56), 5.69 ± 0.28 (R 2 = 0.80, RMSE = 0.43), and 1.20 ± 0.63 min (R 2 = 0.69, RMSE = 0.39), respectively, and the linear model z-value was 5.14 ± 0.39 • C (R 2 = 0.99, RMSE = 0.08). For the same temperatures, the Weibull model t d = 1 values were 20.98 ± 8.8 (R 2 = 0.62, RMSE = 0.46, α (scale parameter) = 2.30, β (shape parameter) = 0.38), 3.84 ± 0.69 (R 2 = 0.85, RMSE = 0.38, α = 1.08, β = 0.66), and 0.87 ± 0.10 min (R 2 = 0.80, RMSE = 0.32, α = 0.22, β = 0.61), respectively and the z-value (using T d = 1 ) was 5.79 ± 0.22 • C (R 2 = 1.0, RMSE = 0.03). A better fit was obtained with the Weibull model for log reductions versus time with higher R 2 and lower RMSE values. Application of AiV inactivation parameters can help reduce the risk of AiV outbreaks.
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