Journal of Applied Microbiology, Vol.121, No.5, 1416-1426, 2016
Survival and expression of acid resistance genes in Shiga toxin-producing Escherichia coli acid adapted in pineapple juice and exposed to synthetic gastric fluid
AimsThe aim of this research was to determine the ability of non-O157 Shiga toxin-producing Escherichia coli (STEC) serogroups to survive with exposure to synthetic gastric fluid (SGF) after adaptation to pineapple juice (PJ) at room and refrigerated temperatures compared to E.coli O157:H7 and to examine the relative transcriptional expression of acid resistance (AR) genes, rpoS, gadA and adiA. Methods and ResultsResistant and sensitive strains belonging to five different STEC serogroups (O26, O103, O104, O111 and O157; n=10) were used in this study. All strains were adapted in PJ (pH 38) stored at 4 and 20 degrees C for 24h, and then the relative transcription levels of genes in all strains were quantified using a real-time quantitative-PCR assay. After adaptation in PJ, the STEC strains were exposed to SGF (pH 15 and 20) at 37 degrees C for 2h. Generally, the STEC adapted in PJ at 4 degrees C displayed enhanced survival compared to acid adaptation in PJ at 20 degrees C and nonadapted controls with exposure to SGF (P<005). Moreover, resistant strains exhibited higher survival rates compared to sensitive strains (P<005). Overall, adaptation at 4 degrees C resulted in significantly (P<005) enhanced gene expression levels in PJ, and transcript levels of gadA were higher than those of the rpoS and adiA genes. ConclusionsThe up-regulation of AR genes due to adaptation in PJ at low temperature may increase STEC survival in acidic environments such as the gastrointestinal tract. Some non-O157 STEC strains, including serotypes O103:H2 and O111:H8, showed relatively high AR levels similar to those of STEC O157:H7. Significance and Impact of the StudyInduction of AR genes in acidic fruit juice, and potentially in other acidic foods may increase the risk of foodborne illness by non-O157 STEC serogroups.
Keywords:acid resistance;gene expression;pineapple juice;real-time quantitative-PCR;Shiga toxin-producing Escherichia coli;synthetic gastric fluid