LuxS modulates motility and secretion of extracellular protease in fish pathogen Vibrio harveyi

In this study, an in-frame deletion of the luxS gene was constructed to reveal the role of LuxS in the physiology and virulence of V. harveyi. The statistical analysis showed no significant differences in the growth ability, biofilm formation, antibiotic susceptibility, virulence by intraperitoneal injection, and the ability of V. harveyi to colonize the spleen and liver of the pearl gentian grouper between the wild-type (WT) and the luxS mutant. However, the deletion of luxS decreased the secretion of extracellular protease, while increased the ability of swimming and swarming. Simultaneously, a luxS-deleted mutant showed overproduction of lateral flagella, and an intact luxS complemented the defect. Since motility is flagella dependent, 16 of V. harveyi flagella biogenesis related genes were selected for further analysis. Based on quantitative real-time reverse transcription-PCR (qRT-PCR), the expression levels of these genes, including the polar flagella genes flaB, flhA, flhF, flhB, flhF, fliS, and flrA and the lateral flagella genes flgA, flgB, fliE, fliF, lafA, lafK, and motY, were significantly up-regulated in the ΔluxS: pMMB207 (ΔluxS+) strain as compared with the V. harveyi 345: pMMB207 (WT+) and C-ΔluxS strains during the early, mid-exponential, and stationary growth phase.

Cell-Free Preparations of Lactobacillus acidophilus Strain La-5 and Bifidobacterium longum Strain NCC2705 Affect Virulence Gene Expression in Campylobacter jejuni

Campylobacter spp. are among the most commonly reported bacterial causes of acute diarrheal disease in humans worldwide. Potential virulence factors include motility, chemotaxis, colonization ability, adhesion to intestinal cells, invasion and epithelial translocation, intracellular survival, and formation of toxins. Probiotic Lactobacillus and Bifidobacterium strains are known to have an inhibitory effect against the growth of various foodborne pathogens. The objective of this study was to investigate the effect of Lactobacillus acidophilus strain La-5 and Bifidobacterium longum strain NCC2705 cell-free spent media (CFSM) on the expression of virulence genes (cadF, cdtB, flaA, and ciaB) of Campylobacter jejuni strain 81-176 and a luxS mutant, using real-time PCR. Our results demonstrated that the CFSM of both probiotic strains were able to down-regulate the expression of ciaB (ratio of −2.80 and −5.51, respectively) and flaA (ratio of −7.00 and −5.13, respectively) in the wild-type Campylobacter strain. In the luxS mutant, where the activated methyl cycle is disrupted, only the ciaB gene (ratio −7.21) was repressed in the presence of La-5 CFSM. A supplementation of homocysteine to restore the disrupted cycle was able to partially reestablish the probiotic effect of both strains. luxS and the activated methyl cycle might play an active role in the modulation of virulence of C. jejuni by probiotic extracts.

Consequences of Disrupting Salmonella AI-2 Signaling on Interactions Within Soft Rots

Within soft rots, Salmonella spp. reach population densities 10- to 100-fold higher than within intact plants. The hypothesis that Salmonella spp. exchange AI-2 signals with Pectobacterium carotovorum to increase its competitive fitness was tested using mutants involved in AI-2 production (luxS) or perception (lsrACDBF or lsrG). Co-infections of a wild-type Salmonella sp. and its AI-2 mutants (at ≈3 to 104) were established in green or red tomato (‘FL 47’ or ‘Campari’ for 3 or 5 days) as well as tomato co-infected with Pectobacterium (at 109) or its luxS mutant. There were no significant differences in the competitive fitness of Salmonella, indicating that AI-2 signaling is not a major input in the interactions between these organisms under the tested conditions. A Salmonella lsrG::tnpR-lacZ resolvase in vivo expression technology (RIVET) reporter, constructed to monitor AI-2-related gene expression, responded strongly to the luxS deletion but only weakly to external sources of AI-2. Growth in soft rots generally decreased RIVET resolution; however, the effect was not correlated to the luxS genotype of the Pectobacterium sp. The results of this study show that AI-2 signaling offers no significant benefit to Salmonella spp. in this model of colonization of tomato or soft rots.