Nosocomial pathogen biofilms on biomaterials: Different growth medium conditions and components of biofilms produced in vitro

Background/Purpose (s): Nosocomial pathogens can develop biofilms on hospital surfaces and medical devices; however, few studies have focused on the evaluation of mono-and dual-species biofilms developed by nosocomial pathogens under different growth conditions. Methods: This study investigated biofilm development by nosocomial pathogens (Acinetobacter baumannii, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa) on biomaterials in different culture media and their components of the extracellular matrix biofilm. Results: The mono-species biofilms showed cell densities from 7.50 to 9.27 Log₁₀ CFU/cm² on natural rubber latex type I (NLTI) and from 7.58 to 8.79 Log₁₀ CFU/cm² on stainless steel (SS). Dual-species biofilms consisted of S. aureus + P. aeruginosa (7.87–8.27 Log₁₀ CFU/cm² in TSBP and TSBME onto SS; p < 0.05), E. coli + P. aeruginosa (8.32–8.86 Log₁₀ CFU/cm² in TSBME onto SS and TSBP onto NLTI; p < 0.05), and S. aureus + E. coli (7.82 Log₁₀ CFU/cm² in TSBME onto SS; p < 0.05). Furthermore, biofilm detachment after proteinase K treatment was 5.54–32.81% compared to 7.95–24.15% after DNase I treatment in the mono-dual species biofilm matrix. Epifluorescence microscopy and scanning electron microscopy (SEM) enabled visualizing the bacteria and extracellular polymeric substances of biofilms on SS and NLTI. Conclusion: Nosocomial pathogens can develop biofilms on biomaterials. Mono-species biofilms of Gram-negative bacteria showed lower densities than dual-species biofilms in TSBME and TSBP. Additionally, dual-species biofilms showed a higher concentration of proteins and eDNA in the extracellular matrix.