Abstract
Cells of Salmonella typhimurium LT2 and other gram-negative bacteria were killed to 10-4-10-7 through freezing with a low concentration (0.5-2.0%) of sodium chloride (NaCl) plus ethanol in cooling rate of 1°C per min to-30°C, and holding at the lowest temperature.The bactericidal effect of freezing was influenced by cooling rate, the lowest temperature, holding time, and the ratio (R) of added NaCl to ethanol. The maximum effect was observed with a solution whose R was 70/30. While, the every concentration of NaCl or ethanol did not change the effect when they had a similar R-value. Proteins and carbohydrates did not influence the cidal effect, but glycerol did.The bactericidal effect of freezing was suggested to be caused by unfrozen solution of NaCl and ethanol which were concentrated through the extracellular slow freezing. The characteristics of ternary solution composed of NaCl, ethanol and water were discussed.
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