Microscopic techniques to evaluate the biofilm formation ability of a marine bacterium Pseudomonas aeruginosa PFL-P1 on different substrata

Biofilm formation in bacteria is strongly affected by the nature of substrata. Different substrata such as glass, polystyrene, steel, ceramic, and rubber were used to assess the biofilm forming ability of a marine bacterium Pseudomonas aeruginosa PFL-P1 using a scanning electron microscope (SEM), atomic force microscope (AFM), and confocal laser scanning microscope (CLSM). The bacterium formed dense biofilms with varied aggregation on different substrata. SEM study revealed small rod-shaped cells with diverse arrangements within the biofilms on all the substrata under study. The AFM study revealed the highest roughness of 545 nm on the ceramic substratum. The biofilms formed on ceramic substratum were characterized with maximum roughness (742 nm), maximum peak height (1,480 nm), and maximum arithmetic mean height (611 nm), significantly higher than all the other substrata (p < .05). AFM studies confirmed that P. aeruginosa PFL-P1 exhibited biofilm heterogeneity on all the substrata. The CLSM study indicated a higher fraction of nucleic acids to alpha-polysaccharides ratio in the biofilms. COMSTAT analysis revealed the highest biofilm biomass of similar to 18 mu m(3)/mu m(2) on the ceramic substratum. The maximum biofilm thickness of similar to 50 mu m in the native state on the ceramic substratum was significantly higher than glass (p = .0015), polystyrene (p = .0001), steel (p = .0035), and rubber substrata (p = .0001). The higher surface roughness of ceramic substratum is accountable for more area for colonization, as evident from higher biomass and thickness of the biofilm. This study provides insight into the substratum properties, which modulate the biofilm forming ability in bacteria.

Automated summary

Biofilm formation in bacteria is influenced by the nature of substrata. A marine bacterium Pseudomonas aeruginosa PFL-P1 formed dense biofilms with varied aggregation on different substrata, with ceramic substratum showing the highest roughness and maximum biofilm thickness. The study provides insight into how substratum properties can impact biofilm formation in bacteria.