A comprehensive comparison of biofilm formation and capsule production for bacterial survival on hospital surfaces

Biofilm formation and capsule production are known microbial strategies used by bacterial pathogens to survive adverse conditions in the hospital environment. The relative importance of these strategies individually is unexplored. This project aims to compare the contributory roles of biofilm formation and capsule production in bacterial survival on hospital surfaces. Representative strains of bacterial species often causing hospital-acquired infections were selected, including Acinetobacter baumannii, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa. The importance of biofilm formation and capsule production on bacterial survival was evaluated by comparing capsule-positive wild-type and capsule-deficient mutant strains, and biofilm and planktonic growth modes respectively, against three adverse hospital conditions, including desiccation, benzalkonium chloride disinfection and ultraviolet (UV) radiation. Bacterial survival was quantitatively assessed using colony-forming unit (CFU) enumeration and the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay and qualitatively by scanning electron microscopy (SEM). Correlations between capsule production and biofilm formation were further investigated. Biofilm formation contributed significantly to bacterial survival on hospital surface simulators, mediating high resistance to desiccation, benzalkonium chloride disinfection and UV radiation. The role of capsule production was minor and species-specific; encapsulated A. baumannii but not K. pneumoniae cells demonstrated slightly increased resistance to desiccation, and neither showed enhanced resistance to benzalkonium chloride. Interestingly, capsule production sensitized K. pneumoniae and A. baumannii to UV radiation. The loss of capsule in K. pneumoniae and A. baumannii enhanced biofilm formation, possibly by increasing cell surface hydrophobicity. In summary, this study confirms the crucial role of biofilm formation in bacterial survival on hospital surfaces. Conversely, encapsulation plays a relatively minor role and may even negatively impact bacterial biofilm formation and hospital survival.