Previous reports have demonstrated that O157 virulence genes, especially the Shiga toxin and LEE–encoded genes, are down-regulated in LB compared to minimal media [38–40]. In addition, presence of trace amounts of GS-4997 mouse glucose has also been shown to down-regulate LEE expression due to catabolite repression and/or acidic pH [38–40]. Hence, the lack of virulence gene
expression in LB in this study conforms to those findings. Experiments with acid-stressed, starved bacteria have shown MI-503 manufacturer that these are likely to be more virulent only on recovery, and over time . Even in minimal media that usually supports O157 virulence gene expression, several of these are suppressed as cultures reach the stationary phase . Butyrate, a key environmental cue in LEE gene expression was limited in the RF used in this study, which may have also caused the LEE suppression . Conditioned media from unrelated cultures have been shown to suppress Shiga toxin gene expression while maintaining O157 growth or suppressing PHA-848125 growth itself [33, 35, 42]. In fact, experimental studies have shown that it is easier to displace O157 in unfiltered rumen fluid versus autoclaved rumen fluid, by addition of “nonfermentable” sugars in the presence of the ruminal microflora . Thus, the
absence of O157 virulence gene expression in RF-preparations may be reflective of the stressful growth environment, suppression due to nutrient limitations, lack of inducers, oxygen deprivation, pH fluctuations and inhibitory metabolites released by resident microbiota. Previous studies have suggested development of acid resistance by Shiga-toxin producing E. coli (STEC) in the rumen as a means for better STEC survival through the ‘stomach-like’ acidic bovine abomasum [43, 44] and have prescribed a role for glutamate-dependent acid resistance system (Gad system) and the tryptophanase (tnaA) enzyme toward this end . Hughes et al., recently demonstrated that O157 LEE expression is down-regulated while the
Gad system is up-regulated in the rumen of cattle . This observation made in animals being fed a grain diet, having a ruminal pH of 5.93, selleck chemical derived a role for the SdiA gene in sensing the acylhomoserine lactone (AHL) signals in the rumen fluid and affecting differential expression of these genes. AHLs formed by ruminal resident flora, are effective only under highly acidic pH and hydrolyze at neutral-alkaline pH [46, 47]. Similarly, the Gad system that relies on the decarboxylation (gadA/B) of glutamate via proton consumption to increase cytoplasmic alkalinity is active at pH 4–4.6 . However, other degradative amino acid decarboxylase and acid-resistance systems are activated in response to low pH (5.2 to 6.9), fermentative-anaerobic growth and stationary phase growth [48, 49] and used more often than the Gad system to counter the deleterious effects of protons.