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Host-like carbohydrates promote bloodstream survival of Vibrio vulnificus in vivo.

Lubin JB, Lewis WG, Gilbert NM, Weimer CM, Almagro-Moreno S, Boyd EF, Lewis AL - Infect. Immun. (2015)

Bottom Line: Sialic acids are found on all vertebrate cell surfaces and are part of a larger class of molecules known as nonulosonic acids.In fact, levels of bacterial persistence in the blood corresponded to the overall levels of these molecules expressed by V. vulnificus isolates.Taken together, these results suggest that molecules similar to sialic acids evolved to facilitate the aquatic lifestyle of V. vulnificus but that their emergence also resulted in a gain of function with life-threatening potential in the human host.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, University of Delaware, Newark, Delaware, USA.

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NulO contributes to biofilm formation and motility. (A) Abiotic biofilm assays were conducted in 96-well polystyrene plates as described in Materials and Methods. At least 2 biological replicates were performed for each experimental condition. The data from the 6-h time point are shown. Error bars represent standard deviations. Statistical evaluation employed one-way ANOVA with the Bonferroni multiple-comparison posttest (P < 0.05). (B and C) Swimming motility was assessed by placing aliquots of WT and isogenic Δnab2 strains onto marine agar plates (0.3%) and measuring zones of motility following 16 h of growth at room temperature. The ΔflhF strain is a mutant of a known flagellar regulator that leads to a nonmotile, aflagellar strain. (D) Transmission electron micrographs of WT CMCP6 and its isogenic Δnab2 strain. (E) Flagellar morphology was assessed by electron microscopy. Several hundred individual bacteria were examined and showed that larger proportions of the Δnab2 population exhibited shortened or missing flagella. Error bars represent standard deviations. At least 2 biological replicates were tested for each experimental condition. One-way ANOVA with Dunnet's posttest was used to examine whether differences between the WT strain and isogenic derivatives were statistically significant. ****, P < 0.0001; ***, P < 0.001; **, P < 0.01 ; *, P < 0.05.
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Figure 3: NulO contributes to biofilm formation and motility. (A) Abiotic biofilm assays were conducted in 96-well polystyrene plates as described in Materials and Methods. At least 2 biological replicates were performed for each experimental condition. The data from the 6-h time point are shown. Error bars represent standard deviations. Statistical evaluation employed one-way ANOVA with the Bonferroni multiple-comparison posttest (P < 0.05). (B and C) Swimming motility was assessed by placing aliquots of WT and isogenic Δnab2 strains onto marine agar plates (0.3%) and measuring zones of motility following 16 h of growth at room temperature. The ΔflhF strain is a mutant of a known flagellar regulator that leads to a nonmotile, aflagellar strain. (D) Transmission electron micrographs of WT CMCP6 and its isogenic Δnab2 strain. (E) Flagellar morphology was assessed by electron microscopy. Several hundred individual bacteria were examined and showed that larger proportions of the Δnab2 population exhibited shortened or missing flagella. Error bars represent standard deviations. At least 2 biological replicates were tested for each experimental condition. One-way ANOVA with Dunnet's posttest was used to examine whether differences between the WT strain and isogenic derivatives were statistically significant. ****, P < 0.0001; ***, P < 0.001; **, P < 0.01 ; *, P < 0.05.

Mentions: Given the common involvement of bacterial surface carbohydrates in biofilm formation (15, 33, 37–39), we next evaluated the potential role of NulO biosynthesis in the ability of V. vulnificus to generate biofilms on abiotic surfaces. The results showed that in the CMCP6 background, nab2 deletion resulted in a defect in biofilm formation compared to that of the WT (Fig. 3A). Moreover, complementation (Δnab2/pnab2 strain) completely restored the ability to form biofilms (Fig. 3A). These data implicate NulO molecules in the generation of V. vulnificus biofilms. However, in contrast to CMCP6, YJ016 (which expresses lower overall levels of NulOs) exhibited minimal, if any, nab2-dependent biofilm formation (not shown).


Host-like carbohydrates promote bloodstream survival of Vibrio vulnificus in vivo.

Lubin JB, Lewis WG, Gilbert NM, Weimer CM, Almagro-Moreno S, Boyd EF, Lewis AL - Infect. Immun. (2015)

NulO contributes to biofilm formation and motility. (A) Abiotic biofilm assays were conducted in 96-well polystyrene plates as described in Materials and Methods. At least 2 biological replicates were performed for each experimental condition. The data from the 6-h time point are shown. Error bars represent standard deviations. Statistical evaluation employed one-way ANOVA with the Bonferroni multiple-comparison posttest (P < 0.05). (B and C) Swimming motility was assessed by placing aliquots of WT and isogenic Δnab2 strains onto marine agar plates (0.3%) and measuring zones of motility following 16 h of growth at room temperature. The ΔflhF strain is a mutant of a known flagellar regulator that leads to a nonmotile, aflagellar strain. (D) Transmission electron micrographs of WT CMCP6 and its isogenic Δnab2 strain. (E) Flagellar morphology was assessed by electron microscopy. Several hundred individual bacteria were examined and showed that larger proportions of the Δnab2 population exhibited shortened or missing flagella. Error bars represent standard deviations. At least 2 biological replicates were tested for each experimental condition. One-way ANOVA with Dunnet's posttest was used to examine whether differences between the WT strain and isogenic derivatives were statistically significant. ****, P < 0.0001; ***, P < 0.001; **, P < 0.01 ; *, P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4496609&req=5

Figure 3: NulO contributes to biofilm formation and motility. (A) Abiotic biofilm assays were conducted in 96-well polystyrene plates as described in Materials and Methods. At least 2 biological replicates were performed for each experimental condition. The data from the 6-h time point are shown. Error bars represent standard deviations. Statistical evaluation employed one-way ANOVA with the Bonferroni multiple-comparison posttest (P < 0.05). (B and C) Swimming motility was assessed by placing aliquots of WT and isogenic Δnab2 strains onto marine agar plates (0.3%) and measuring zones of motility following 16 h of growth at room temperature. The ΔflhF strain is a mutant of a known flagellar regulator that leads to a nonmotile, aflagellar strain. (D) Transmission electron micrographs of WT CMCP6 and its isogenic Δnab2 strain. (E) Flagellar morphology was assessed by electron microscopy. Several hundred individual bacteria were examined and showed that larger proportions of the Δnab2 population exhibited shortened or missing flagella. Error bars represent standard deviations. At least 2 biological replicates were tested for each experimental condition. One-way ANOVA with Dunnet's posttest was used to examine whether differences between the WT strain and isogenic derivatives were statistically significant. ****, P < 0.0001; ***, P < 0.001; **, P < 0.01 ; *, P < 0.05.
Mentions: Given the common involvement of bacterial surface carbohydrates in biofilm formation (15, 33, 37–39), we next evaluated the potential role of NulO biosynthesis in the ability of V. vulnificus to generate biofilms on abiotic surfaces. The results showed that in the CMCP6 background, nab2 deletion resulted in a defect in biofilm formation compared to that of the WT (Fig. 3A). Moreover, complementation (Δnab2/pnab2 strain) completely restored the ability to form biofilms (Fig. 3A). These data implicate NulO molecules in the generation of V. vulnificus biofilms. However, in contrast to CMCP6, YJ016 (which expresses lower overall levels of NulOs) exhibited minimal, if any, nab2-dependent biofilm formation (not shown).

Bottom Line: Sialic acids are found on all vertebrate cell surfaces and are part of a larger class of molecules known as nonulosonic acids.In fact, levels of bacterial persistence in the blood corresponded to the overall levels of these molecules expressed by V. vulnificus isolates.Taken together, these results suggest that molecules similar to sialic acids evolved to facilitate the aquatic lifestyle of V. vulnificus but that their emergence also resulted in a gain of function with life-threatening potential in the human host.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, University of Delaware, Newark, Delaware, USA.

Show MeSH
Related in: MedlinePlus