title: Antibiotic additive and synergistic action of rutin, morin and quercetin against methicillin resistant Staphylococcus aureus.
abstract: BACKGROUND
  To determine the effect of flavonoids in conjunction with antibiotics in methicillin resistant Staphylococcus aureus (MRSA) a study was designed. The flavonoids included Rutin, Morin, Qurecetin while antibiotics included ampicillin, amoxicillin, cefixime, ceftriaxone, vancomycin, methicillin, cephradine, erythromycin, imipenem, sulphamethoxazole/trimethoprim, ciprofloxacin and levolfloxacin. Test antibiotics were mostly found resistant with only Imipenem and Erythromycin found to be sensitive against 100 MRSA clinical isolates and S. aureus (ATCC 43300). The flavonoids were tested alone and also in different combinations with selected antibiotics.
  
  
  METHODS
  Antibiotics and flavonoids sensitivity assays were carried using disk diffusion method. The combinations found to be effective were sifted through MIC assays by broth macro dilution method. Exact MICs were determined using an incremental increase approach. Fractional inhibitory concentration indices (FICI) were determined to evaluate relationship between antibiotics and flavonoids is synergistic or additive. Potassium release was measured to determine the effect of antibiotic-flavonoids combinations on the cytoplasmic membrane of test bacteria.
  
  
  RESULTS
  Antibiotic and flavonoids screening assays indicated activity of flavanoids against test bacteria. The inhibitory zones increased when test flavonoids were combined with antibiotics facing resistance. MICs of test antibiotics and flavonoids reduced when they were combined. Quercetin was the most effective flavonoid (MIC 260 μg/ml) while morin + rutin + quercetin combination proved most efficient with MIC of 280 + 280 + 140 μg/ml. Quercetin + morin + rutin with amoxicillin, ampicillin, cephradine, ceftriaxone, imipenem, and methicillin showed synergism, while additive relationship was indicated between morin + rutin and amoxicillin, cephradine, ceftriaxone, imipenem, and methicillin. Quercetin alone had an additive effect with ampicillin, cephradine, ceftriaxone, imipenem, and methicillin. Potassium leakage was highest for morin + rutin + quercetin that improved further in combination with imipenem. Morin and rutin alone had no activity but in combination showed activity against test bacteria.
  
  
  CONCLUSIONS
  The flavonoids when used in combination with antibiotics were found to increase each other activity against test bacteria. The relationship between the flavonoids and antibiotics in most of the cases was additive. However in a few cases synergism was also observed. Flavonoids alone or in combinations also damaged bacterial cell membrane.
authors: Department of Microbiology, Kohat University of Science and Technology, Kohat, KP, Pakistan. pharmacyst2000@yahoo.com.
authors: Muhammad Usman
authors: MU
authors: Amin
authors: Department of Pharmacy, Shaheed Benazir Bhutto University, Sheringal, Dir Upper, KP, Pakistan. pharmacistkhurram@hotmail.com.
authors: Muhammad
authors: M
authors: Khurram
authors: Department of Microbiology, Kohat University of Science and Technology, Kohat, KP, Pakistan. baharkk75@yahoo.com.
authors: Baharullah
authors: B
authors: Khattak
authors: Department of Microbiology, Kohat University of Science and Technology, Kohat, KP, Pakistan. jafarkhan1@yahoo.com.
authors: Jafar
authors: J
authors: Khan
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004351:Drug Resistance / Q000187:drug effects*; D004357:Drug Synergism; D005419:Flavonoids / Q000494:pharmacology*; D006801:Humans; D008712:Methicillin / Q000494:pharmacology; D055624:Methicillin-Resistant Staphylococcus aureus / Q000187:drug effects*; D008826:Microbial Sensitivity Tests; D011794:Quercetin / Q000494:pharmacology*; D012431:Rutin / Q000494:pharmacology*; D047309:Flavones
chemical_list: D000900:Anti-Bacterial Agents; D005419:Flavonoids; D008712:Methicillin; D011794:Quercetin; D012431:Rutin; C008548:morin; D047309:Flavones