Determination of optimum incubation time for formation of Pseudomonas aeruginosa and Streptococcus pyogenes biofilms in microtiter plate

Abstract

Pseudomonas aeruginosa and Streptococcus pyogenes are the most common pathogens to humans and are able to form a biofilm following ineffective precautionary approach. Biofilm is defined as a surface-attached community of bacterium embedded in an extracellular matrix which leads to tremendous problems in the environment, among humans and animals. This study aims to investigate the ability of P. aeruginosa and S. pyogenes to form biofilms in 96-well plate before further study in antibiofilm will be done. Initially, the 96-well plate was added with 100 μl of overnight P. aeruginosa culture with optical density (OD) 0.1 and S. pyogenes culture with OD 0.05. The cultures were incubated for 7 days at 37°C to justify the formation of biofilm. Subsequently, stained blue biofilm was detached from the plate by using 95% ethanol. Biofilms were finally measured using a micro plate reader at 570 nm and were classified based on the adherence strength formula. P. aeruginosa and S. pyogenes biofilms strongly adhered to the plates on days three, four, five and six. Interestingly on day three, biofilms showed the highest formation. However, moderate biofilm formation onto the plates by both P. aeruginosa and S. pyogenes were observed on day two, but non-adherence was observed on days one and seven. Day three is the optimum cultivation period for P. aeruginosa and S. pyogenes to switch into a strong biofilm in microtiter plate and could be beneficial for antibiofilm experiments.

Viability of Salmonella Typhimurium biofilms on major food-contact surfaces and eggshell treated during 35 days with and without water storage at room temperature

“…Typhimurium biofilm on the ES surface ( Kim et al., 2016 ). Similarly, it has been suggested that wood encourages biofilm formation because of its porosity and absorbency, which can entrap organic material and bacteria ( Adetunji and Isola, 2011 ; Al-kafaween et al., 2019 ). Moreover, moisture retention on the ES could also account for the greater viability of S. Typhimurium at low temperatures because the eggs were stored while still wet ( Rizk et al., 1966 ).…”

Evaluation of the effect of different growth media and incubation time on the suitability of biofilm formation by Pseudomonas aeruginosa and Streptococcus pyogenes

“…Previous studies have indicated that temperature, nutrients and other components in media affected the attachment of microorganisms to the surfaces of various materials (Hood and Zottola, 1997;Iversen et al, 2004). Study by (Al-kafaween et al, 2019b) showed that the E. coli produced biofilm on day 3 and day 5 in Brain heart infusion (BHI) and Luria broth (LB) at OD 0.05 and OD 0.1 (Al-kafaween et al, 2019). Studies have used TSB for Staphylococcus and Pseudomonas species but this depends on growing condition of microorganisms and optical density (Nyenje et al, 2013;Stepanovi et al, 2000).…”

“…or nonadherent [NA = OD ODC)]. ODC was referred to the density of the negative control (Al-kafaween et al, 2019;Al-kafaween et al, 2019a;Nyenje et al, 2013).…”

New Paradigm in Explaining Dry Socket: Case Studies

“…1-The onset: clinical symptoms culminate in up to 4 days, and mainly manifested through severe pain, this period is equivalent and just refer to the incubation period of Pseudomonas aeruginosa [8].…”