FIG 2 .

Capsular polysaccharide (CPS) production by V. vulnificus under heat shock conditions. (A) Colony morphology. Wild-type and wbpP mutant strains of V. vulnificus were grown on LBS agar plates for 12 h at 30°C and 42°C, and then the opacity of the bacterial colonies was examined. "O" and "T" indicate the colonies with opaque and transparent morphologies, respectively. (B) Polyacrylamide gel electrophoresis of the CPS extracts. CPS preparations extracted from each strain grown at 30°C and 42°C for 48 h were analyzed by electrophoresis in a 5% stacking polyacrylamide gel and subsequent staining with Stains-All. (C) Quantification of the CPS contents. Concentrations of the extracted CPS were quantified using d-galacturonic acid as a standard as described by Taylor (53). The estimated concentrations were expressed as nanograms of galacturonic acid equivalents in 1 μl of CPS extract. (D and E) Dispersal of biofilms. (D) Biofilms formed by wild-type V. vulnificus in 2 ml AB-fumarate broth at 30°C (labeled "1") were further incubated in 4 ml PBS at 30°C or 42°C to allow the second biofilms (labeled "2" and "4") to be formed by the cells dispersed from the first biofilms. The remaining first biofilms (labeled "3" and "5") and newly formed second biofilms around the air-liquid interfaces were stained with crystal violet. (E) The amounts of crystal violet were estimated by spectrophotometry at 550 nm.