Impact of routine laboratory culture media on in-vitro biofilm formation of Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecalis

Objectives: This study was aimed to determine the efficacy of four routine laboratory culture media onbiofilm formation of Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus feacalis.Methods: A sterile flat bottom 96 well plate was inoculated using 0.5 McFarland equivalent standardcell suspension of P. aeruginosa, S. aureus and E. feacalis and the growth rate of planktonic cells wasquantified by measuring the optical density (OD492) at two hour intervals. Influence of culture mediumon adhesion of bacteria as an initial step of biofilm formation in the presence of four culture media(Nutrient broth (NB), Brain Heart Infusion (BHI) broth, Luria-Bertani (LB) broth and RPMI 1640) wasquantified using MTT (3-[4, 5- dimethylthiazole-2-yl]-2, 5- diphenyltetrazolium bromide) assay after90 minutes adhesion. Biofilms of P. aeruginosa, S. aureus, E. feacalis and their 1:1 mixed biofilmswere developed and the growth was quantified using MTT metabolic activity at 24 hour time intervals.Scanning electron microscopy (SEM) was performed to assess the ultrastructure.Results: On comparing the relative growth of the bacteria in different culture media, the maximumgrowth of all three planktonic cultures was achieved using BHI broth. All mono species and mixedspecies cultures exhibited their maximum adhesion in the presence of RPMI 1640. All biofilm exhibitedthe maximum growth in BHI broth. SEM imaging had shown the enhanced growth of ultrastructure ofthe biofilm with the presence of BHI broth.Conclusions: The maximum planktonic and biofilm growth was achieved with BHI broth. However,bacterial adhesion was enhanced in the presence of RPMI 1640