Anti-biofilm activity of Coumarin compound and evaluate effect on the gene expression of icaA and cifA genes in Staphylococcus aureus

Abstract

The aim of this work is to examine the anti-biofilm capabilities of coumarin compound and apply them to specific virulence genes that contribute to the formation of biofilm in multidrug-resistant Staphylococcus aureus. The lab. of the UOB Institute of Genetic Eng. and Biotech.provided Twenty-five isolates of S. aureus. Ten isolates of S. aureus that were resistant to several drugs were chosen, and their capacity to produce biofilm was examined using the micro-titer plate method. The lab. of the UOB Institute of Genetic Eng. and Biotech. provided these isolates. By growing the isolates on Mannitol salt agar and applying the VITEK-2 technology, the diagnosis was verified. According to the findings, every isolate had a high capacity for biofilm production. Furthermore, a number of tests were carried out on the coumarin compound, such as determining the total phenolic content, assessing the antioxidant activity, evaluating the biofilm formation activity, and comparing isolates treated with a coumarin substance with isolates that were not in order to analyse the expression of the icaA and cifA genes. The findings demonstrated a progressive rise in the Coumarin compound's total phenolic content as concentration rose, with significant differences (P≤0.01), the highest values were 72.24 mg/g in 100 mg/ml. Furthermore, the findings showed that when compound concentrations rose, the scavenging activity (antioxidant activity) gradually increased. The activity of coumarin gradually increases from 29.58 at 0.312 mg/ml to 63.50 at 10 mg/ml. The activity of the natural antioxidant (vitamin C) increased sharply at lower concentrations (from 72.72 at 0.312 mg/ml to 90.43 at 2.5 mg/ml), but at higher concentrations, the rate of increase slowed and eventually reached a constant value of 92.22 at 10 mg/ml. Similarly, the artificial antioxidant (BHT) increased in activity as concentration increased, starting at 32.66 at 0.312 mg/mL and reaching 90.12 at 10 mg/ml. The study's findings demonstrated that the coumarin blocked 100% of S. aureus biofilm development at 8 mg/ml and decreased it at 2 mg/ml. The investigation revealed that all isolates possessed the virulence genes icaA and cifA, which are in charge of biofilm formation in S. aureus. Additionally, compared to the isolates that were left untreated, the gene expression data revealed lower levels of icaA and cifA following coumarin treatment.