ARS | Publication request: Bioenergetic Confirmation of Viable Pathogens in Foods by Atp-Bioluminescence

Submitted to: Meeting Abstract
Publication Acceptance Date: May 21, 2004
Publication Date: August 1, 2004
Citation: Tu, S. 2004. Bioenergetic Confirmation Of Viable Pathogens In Foods By Atp-Bioluminescence. Meeting Abstract. Luminescence 19:182, 2004.

Technical Abstract: The cellular ATP content of fourteen freshly harvested bacteria including Bacillus, Campylobacter, Citrobacter, Escherichia, Lactobacillus, Listeria, Pediococcus, Pseudomonas, Salmonella, Streptococcus and Yersinia, was determined using a luciferin-luciferase bioluminescence approach. Incubation of bacteria with carbonyl cyanide meta-chlorophenyl hydrazone (CCCP), a membrane protonophore, prior to cell breakage substantially lowered the bioluminescence signals indicating a decrease of cellular ATP content. The addition of CCCP after cell breakage had no detectable effect on the ATP levels. This differential effect of CCCP was not observed using heat-killed bacteria, i.e., the ATP content was not affected by CCCP incubation. The CCCP effects on cellular ATP level were detectable in bacterial suspensions with 10(3) to 10(6) CFU/mL. This procedure was applied to detect E. coli O157:H7 spiked in ground beef. After a brief enrichment for about 6 h at 37 degrees C, the E. coli cells were captured and concentrated by specific immunomagnetic beads. The captured E. coli cells were then subject to the CCCP treatment. The results demonstrated that the presence of less than one CFU of viable E. coli O157:H7 per gram of ground beef could be detected.