Study the Effect of Alchemilla vulgaris on Biofilm Production by Staphylococcus Aureus That Isolated from Wounds and Respiratory Tract of Patients

Abstract

Alchemilla vulgaris, an aggregate species commonly called lion's foot or Lady's mantle, is a perennial herbaceous plant from the Rosaceae family. Clinical specimens were cultured, including wound swabs, sputum, and throat swabs, and six isolates were identified as Staphylococcus aureus by colony morphology, Gram stain, cultivation on mannitol salt agar, coagulase test, and a set of biochemical tests, three isolates from wound swabs and three from the respiratory tract (sputum and throat swabs).  Alchemilla vulgaris(L.) seeds ethanol extract was extracted by 70% ethanol, and aqueous extract by Soxhlet, then dried by lyophilizer. Phytochemical investigation was performed using petroleum ether, ethyl acetate, and 70% ethanol. Preliminary phytochemical examination showed the presence of alkaloids, flavonoids, phenols, glycosides, tannins, and saponins in both alcoholic and aqueous extracts of Alchemilla vulgaris, with more concentration in the alcoholic extract than in the aqueous extract. Antimicrobial sensitivity test was done by disk diffusion method using six antibiotics, and most isolates were multidrug resistant to oxacillin and gentamicin, while ciprofloxacin and levofloxacin showed high activity. Methanol extract shows more activity than aqueous extract, and Staph. aureus from wound sources were more susceptible than those isolated from the respiratory tract. Inhibition zone ranged from 20- 11 mm in diameter for methanol extract and 16- 8 mm in diameter for aqueous extract. MIC was 100 mg/ mL. Biofilm was detected by two methods, Congo red agar and microtiter plate assay reveal strong biofilm formation for all isolates. Antibiofilm activity was examined, and both ethanol and aqueous extracts decreased the bacterial biofilm formation.