ABSTRACT
This research aimed to evaluate the effect of acid stress on the expression of stress regulator (grxB and rpoS) and virulence (ompA, hfq, and cpa) genes of Cronobacter sakazakii Yrt2a. The results showed that C. sakazakii Yrt2a experienced decrease in number during acid stress and was no longer culturable 90 min post exposure to pH 3.0. During acid stress, the expression of grxB, rpoS, ompA, cpa and hfq was upregulated by 2.15; 2.19; 1.55; 1.1 and 1.41 log, respectively. However, all genes expression was downregulated when the bacteria entered the unculturable state. The expression of gene grxB, rpoS, ompA, cpa decreased to 1.04; 0.37; 0.84 and 1.71 log, respectively; while hfq gene expression reached a level lower than that of control. This research implies a supposition that during acid stress, C. sakazakii was capable of maintaining its culturability and pathogenicity until they are no longer culturable.
ABSTRACT
Indonesian kefir grains are potential sources of lactic acid bacteria (LAB) that may act as probiotics with specific functional properties. In this study we explored the potential of the probiotic and cholesterol-lowering effect of LAB isolated from Indonesian kefir grains obtained from Bogor, Bandung, Jakarta, and Yogyakarta. The results revealed that 10 isolates showed considerable survivability at low pH and bile salt with total cell reduction of ~3 log colony-forming units per milliliter after exposure to pH 2.5 and 0.5% (w/v) bile salt for 1 and 3 h, respectively. All strains exhibited strong antimicrobial activities against pathogenic bacteria and were sensitive to a wide spectrum of antibiotics but exhibited weak bile salt hydrolase activity. Identification based on 16S RNA suggested that nine isolates were Lactobacillus kefiri and one was Lactobacillus rhamnosus. The ability of the isolates to reduce cholesterol from the media varied, ranging from 22.08% to 68.75% with the highest reduction shown by L. kefiri JK17. The ability to remove cholesterol from the media decreased greatly in resting and dead cells, ranging from 14.58% to 22.08% in resting cells and from 7.89% to 18.17% in dead cells. It can be concluded that Indonesian kefir grains contain LAB potentially acting as probiotics capable of reducing cholesterol. The cholesterol-lowering effect especially occurs when the cells are metabolically active.
Subject(s)
Cholesterol/metabolism , Kefir/microbiology , Lacticaseibacillus rhamnosus/isolation & purification , Lactobacillus/isolation & purification , Probiotics/isolation & purification , Probiotics/metabolism , Anti-Bacterial Agents , Antibiosis , Bile Acids and Salts , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Hydrogen-Ion Concentration , Hydrolases/metabolism , Indonesia , Lactobacillus/classification , Lactobacillus/genetics , Lactobacillus/metabolism , Lacticaseibacillus rhamnosus/classification , Lacticaseibacillus rhamnosus/genetics , Lacticaseibacillus rhamnosus/metabolism , Microbial Viability/drug effects , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
Cronobacter spp. in powdered infant formula has been etiologically linked to meningitis and necrotizing enterocolitis in certain groups of infants. This study aimed to determine whether C. sakazakii Yrt2a strain experiencing desiccation stress could enter viable but nonculturable (VBNC) state as well as to examine the expression of genes associated with stress and virulence during the above states. Stress and VBNC conditions were determined based on viability and culturability assays. Expression of genes related to stress (rpoS) and virulence (hfq and ompA) was evaluated by real-time PCR. The results showed that C. sakazakii Yrt2a entered VBNC 24 days post exposure to 2 h of desiccation treatment. The expression of rpoS, hfq and ompA genes was up-regulated during stress conditions, suggesting that Cronobacter successfully managed stress to maintain its culturability while maintaining its virulence. The expression of the target genes decreased at VBNC state but remained higher than that of a normal state. These findings reinforce the assumption that C. sakazakii undergoing VBNC state maintains its pathogenicity.
