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Adaptability to Various Growth Conditions of Biofilm Associated Extended-Spectrum-Beta-Lactamases Producing Bacteria

Sinan Baho, Ruth Reid and Shivanthi Samarasinghe

Extended-Spectrum β-Lactamase (ESBL) producing bacteria are becoming increasingly prevalent in biofilmassociated infections. Bacteria form biofilms that allow their survival in hostile environments. The amount of formed biofilm is affected by external environmental factors. This study investigates the effect of specific parameters (media type, incubation condition, and growth stage) on the amount of produced biofilm on antibiotic resistant bacterial strains, Escherichia coli (CTX-M-15, TEM-3, and IMP-type) and Klebsiella pneumoniae (OXA-48, SHV-18, NDM-1, and KPC-3). The amount of biofilm formed was measured at different time points (6, 12, 24 and 48 h) of incubations under static and shaking conditions, using three different types of media (nutrient broth, LB broth, and AB broth). Statistical tests showed that there was a significant difference in biofilm level (p<0.01) for 64 out of 80 tests (80%) when grown under different types of media. Growing under different incubation conditions also showed a statistical difference in biofilm level (p<0.05) for 76 out of 120 tests (63%). Stage of growth of the same species also showed statistical difference, 20 out of 24 tests (83%) for E. coli and 24 out of 24 tests (100%) for K. pneumoniae. These findings suggested that biofilm formation is highly affected by incubation conditions, strains’ stage of growth, and media type demonstrating that these conditions may play a role in adaptability of the ESBLs on different environmental conditions and their increased prevalence in biofilm associated infections.