Phytochemical analysis and antibacterial activity of some medicinal plants against methicillin-resistant Staphylococcus aureues

Tlre objective of this study is, first to report the prevalence of methicillin resistant Staphylococcus aureues (MRSA) infections and their antibiotic susceptibility patterns in Teaching and Emergency Hospital in Erbil city, and second to screen four medicinal plants: Eucalyptus globulus, Anethunt graveolens, Prosopis farcta and Ficus carica (both aqueous extract and ethanolic extract) for potential antibacterial activity against MRSA isolates, then screening the phytochemical properlies of these plants. Out of total 1 10 strains of Staphylococcus aureues isolated from different clinical specimens, \5 (13.6%) were found to be methicillin resistant. All MRSA strains were found to be resistant to penicillin and gentamycin while 86.66% were resistant to cephalothin ,doxycyciin and tobramycin. However, no strains were resistant to vancomycin. Determination of rninimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values were performed using micro broth dilution method. The MIC/\,IBC of aqueous and ethanolic extracts of E. gtobulzus against MRSA isolates were 25112.5 mg/ml and 6.2513.125 mg/ml respectively, while of P. farcta werc 100125 mg/ml and 25112.5 mg/ml respectively. The results also revealed that extracts of F. carcia were inactive against MRSA. Preliminary phytochemical analysis revealed the presence of tannins and phenols in almost all the crude extracts followed by alkaloids and resins, while the glycosides was not detected in any extracts of the four plants tested.

and hospital acquired infections.It can cause a range of infectious disease from mild condltions, such'as soft tissue infections, to severe life threatening debiliation such as endocarditic to death (1,2).Methicillin- resistant strains of staphylococci were identified immediately upon the introduction of methicillin into clinical practice (3).The f,rrst outbreak caused by MRSA occurred in a British hosnital in the early 1960s and emerged in the United States in the mild 1980s (1,4).MRSA strains have become frequently resistant to multiple other antimicrobial agents and many investigators have reported an increase in the incidence of MRSA during recenr years (5. 6, 7).
Interest in plants as source of antinlicrobial agents is growing.This is because of that the plant-derived medicines have been part of traditional health care in most parts of the world and because of the side effects and the resistance that pathogenic organisms build against antibiotic and there has been an alarming increase in the incidence of new and reemerging infectious diseases (8,9), and because the antimicrobial properties of medicinal plants are well documented in different parts of the world (10, 11,I2).
Eucalyptus species belong to the family Myrtaceae.It is a large genus of aromatic trees which are distributed wideiy throughout the world and are famous for their rapid growth (13, l4).
The oils of the Eucalyptus plants are used frecluently as a remedy for cold and cough.They are used in pharmaceuficals such as cough syrups, lozenges, nasal drops and mouthwash.Eucctlypttts is an ingredient in over-thecounter pharmaceuticals as temporary relief of minor aches and pains of muscles and has been employed in steam inhaiation treatments and treatment of wounds (15).Aqueous and organic extracts of the leaves of Eucalyptus species were investigated and the results show anti-inflammatory, antifungal, antibacterial activity against some gram positive and negative bacteria and may also provide potent antiseptic properties (lZ, 13, l6 Prosopis farcta is a small prickly spiny trees or shrubs.In its native range, it is wide spread and a weed of wheat and cotton fields, invading by root suckers. It is native to Northern Africa, south Western Asia, United States, Irac1, Kuwait, Iran and Turkey (14,21).The antimicrobial activity of extracts from Prosopis spp. was investigated and the results show antifungal, antibacterial activity against some gram positive and negative bacteria, antidiarrheic, antiparasitic (22,23,24).Ficus ts a genus of about 850 species of woody trees, shrubs, or small tree in the family diarrhea, toothache and microbiai infections (11).Various pharmacological actions such as antifungal, antibacterial, antioxidant and anti-inflammatory have been described for F.icus spp. (26,27, Z8).The aim of this study is to determine the prevalence of MRSA infection and their susceptibility pattern to various antimicrobial agents and to screen four medicinal plants for their potential antibacterial activity of Erbil Province on clinical'isolates of MRSA and then screening of these plants for their phytochemical compounds.  1.5rng/ml were recorded for ethanol exftact and 100mg/m1 and 25mglml for aqueous extract for P. farcta respectively.Also MIC and MBC values of 5Omg/ml and 25mglml and 12.5mg/ml and 12.5mg/ml were obtained for aqueous and ethanol extract of A. graveolens respectively.

Materials and methods
Only'extracts of F. carica were inactive against MRSA.The results of the preliminary phyochemical analysis are shown in Table (5).

Discussion
The prevalence rate of MRSA infection in the present study was found to be \3.60/0,which is in accordance with other reports (5,33).On the contrary, some of the reports show an alarmingly high incidence of MRSA infection.
Rijal and coworker demonstrate an increase in methicillin resistant Staphylococ.cttsaureues (56.14%) in patients visiting Western Regional Hospital in Pokhara (34), and Vidhani and colleagues found 51.6% of MRSA in Orthopaedics and Burn Units of LN Hospital, New Delhi (6).The SENTRY antimicrobial surveillance programme found that the prevalence of MRSA in hospital between 1997 and 1999 were very high in the countries like in Japan (71.6%), Singapore (62.9%),Taiwan (61.1%),Portugal (54.4%) and Australia (23.6%) (35).A11 MRSA isolates encountered in this study were completely resistant to antibiotics such as penicillin, gentamycin and oxacillin.A similar resr.rlt was noted for peniciiiin and oxacillin among MRSA strains from India, Nepal, Medagascor and Saudi Arabia (36,37,38,39).All MRSA isolates in this study were completely sensitive to vancomycin.A simiiar result was obtained for vancomycin in previous reports (7,38).
Staphylococcus aureues is a versatile human pa'thogen, it was strongly considered as a major cause of nosocomial infection.In recent years, the prevalence of MRSA has increased worldwide as it is evident from many surveillance studies (35, 40, 4l).
However, infection with MRSA varies widely from one geographic location to another, from hospital to hospital and over.time (42,43) aminoglycosides, tetracyclin, anc miscellaneous group, and this is probably due to the indiscriminate and empirical use of these drugs (2, 44 Umbelliferae, to which the plants used in the present study belong.The inhibitory effects of these medicinal plants on the microorganisms may therefore, be due to the presence of above phyochemical components (16).
Different plants possess different constituents and in different concentrations, which accounts for differential antimicrobial effects, as also suggested earlier (8).The mechanisms thought to be responsible for these phltochemicals against microorganisms vary and depend on above compounds.
Their mechanism of actions may include enzyme inhibition by the oxidized compounds and act as a source of stable free radical and often leading to inactivation of the protein and loss of fi rncrion Thev have the ahilitv to complex with extracellular and soluble proteins and to complex with bacterial cell walis and disrupt microbial membranes' (47, 50).
Tikrit Journal of Pharmaceutical Sciences 2009 S(2):lI6-126 Introduction Staphylococcus aureues is one of the most significant human pathogen ).
They are native to Southwest Asia' and the Eastem Mediterranean region (from Greece to Pakistan).The fruit of most species are edible though they are usualiy of only local economic importance or eaten as bush food (17,25).Varior:s parts of this plant like bark, leaves, tend.ershoots, fruits, seeds and latex are medicinally important.Ficus snn.has heen annlied in folk medicine for cough, sore throat, burning, Tikrit Journal of Pltarmaceuticul Sciences 2009 5(2):116-126 Moraceae.

Table Emergency
Hospital in Erbil city for various bacteriological examination.for aqueous and ethanol extracts of E. globulus respectively, while the corresponding MBC values are 12.5mg/m1 and 3.125mglml.The MIC and MBC values of 25mglrnl and obtained Phenols and tanninswere present in almost all the crude plant extracts (aqueous and ethanol) followed by alkaloids and resins, and

Table ( 4
): Minimum inhibitory and bactericidal concentration of plant extracts on MRSA.
). was strongly active against MRSA strains with ethanol extracts than water extract (MIC value was 6.25mglml and MBC was 3.125mglml).