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A by-product of glucose produced during sterilization (121°C, 15 lb/in2 , 15 min) at neutral pH and in the presence of phosphate (i.e., phosphate-buffered saline) was bactericidal toEscherichia coli O157:H7 (ATCC 43895). Other six-carbon (fructose and galactose) and five-carbon (arabinose, ribose, and xylose) reducing sugars also produced a toxic by-product under the same conditions. Fructose and the five-carbon sugars yielded the most bactericidal activity. Glucose concentrations of 1% (wt/vol) resulted in a 99.9% decline in the CFU of stationary-phase cells per milliliter in 2 days at 25°C. AnrpoS mutant (pRR10::rpoS ) of strain 43895 (FRIK 816-3) was significantly (P &lt; 0.001) more sensitive to the glucose-phosphate by-product than the parent strain, as glucose concentrations from 0.05 to 0.25% resulted in a 2- to 3-log10 reduction in CFU per milliliter in 2 days at 25°C. Likewise, log-phase cells of the wild-type strain, 43895, were significantly more sensitive (P &lt; 0.001) to the glucose-phosphate by-product than were stationary-phase cells, which is consistent with the stability ofrpoS and the regulation ofrpoS -regulated genes. The bactericidal effect of the glucose-phosphate by-product was reduced when strains ATCC 43895 and FRIK 816-3 were incubated at a low temperature (4°C). Also, growth in glucose-free medium (i.e., nutrient broth) did not alleviate the sensitivity to the glucose-phosphate by-product and excludes the possibility of substrate-accelerated death as the cause of the bactericidal effect observed. The glucose-phosphate by-product was also bactericidal toSalmonella typhimurium ,Shigella dysenteriae , and aKlebsiella sp. Attempts to identify the glucose-phosphate by-product were unsuccessful. These studies demonstrate the production of a glucose-phosphate by-product bactericidal toE. coli O157:H7 and the protective effects afforded byrpoS -regulated gene products. Additionally, the detection of sublethally injured bacteria may be compromised by the presence of this by-product in recovery media.

applied and environmental microbiology

Lethality of a Heat- and Phosphate-Catalyzed Glucose By-Product to <i>Escherichia coli</i> O157:H7 and Partial Protection Conferred by the <i>rpoS</i> Regulon

Lethality of a Heat- and Phosphate-Catalyzed Glucose By-Product to Escherichia coli O157:H7 and Partial Protection Conferred by the rpoS Regulon