Growth Characteristics of Vibrio parahaemolyticus Biofilm on Glass Surfaces and Biofilm Detachment under Ultrasound Treatment
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Abstract:
The growth characteristics of Vibrio parahaemolyticus biofilms on glass surfaces, the effects of environmental factors during biofilm formation, and biofilm detachment under ultrasound treatment were investigated. The morphology of V. parahaemolyticus biofilms on the glass surface was observed by crystal violet staining, biofilm biomass was measured using an enzyme-linked immunosorbent assay (ELISA) plate reader at 595 nm, and the extent of biofilm detachment was measured by counting colony-forming units (CFU) using the plate count method. V. parahaemolyticus biofilms formed on the glass surface, and could be directly observed using crystal violet staining. The reticular structure formed by the V. parahaemolyticus biofilm became denser with increasing cultivation time. The optimum parameters for V. parahaemolyticus biofilm formation with respect to biomass were determined using the plate reader. For a culture medium salinity of 3%, a rotation speed of 70 r/min, and an incubation time of 24 h, the number of bacteria in the mature V. parahaemolyticus biofilm was 2.56 × 107 CFU/cm2. Optimal biofilm detachment was obtained under pulsed ultrasound treatment (30-s sonication at intervals of 30 s) applying a frequency at 50 kHz for four minutes, and the viability of bacteria was maintained under these conditions.
