ANTIMICROBIAL ACTIVITY OF AQUEOUS & METHANOLIC EXTRACT OF YOUNG & MATURE LEAVES OF PSIDIUM GUAJAVA L. (GUAVA)

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1 J. Indian bt. Sc. Vl. 92 (1&2) 2013 : 4751 ANTIMICROBIAL ACTIVITY OF AQUEOUS & METHANOLIC EXTRACT OF YOUNG & MATURE LEAVES OF PSIDIUM GUAJAVA L. (GUAVA) MEENAKSHI VAIDYA Department f Btany, Mithibai Cllege, Vile Parle (West), Mumbai minakshispv@gmail.cm Psidium guajava cmmnly called as guava belngs t family Myrtaceae. The leaves have been used in flk medicine as remedy fr diarrhea and als fr its antimicrbial prperties. In the present study a cmparative analysis f aqueus and methanlic extracts was dne fr bth yung and mature leaves. Phytchemical screening was carried ut by standard bichemical tests, and t evaluate the pssibility f nvel pharmaceuticals having antimicrbial ptential. The micrbial assay was dne by agar cup methd and MIC against the fllwing micrrganisms, and. Key Wrds: Psidium guajava, yung and mature leaves, antimicrbial, MIC Natural plants being used lcally as primary health remedies, due t their pharmaclgical prperties, is cmmn in many cuntries including India. Psidium guajava Linn.is als knwn as (guava), belngs t the family Myrtaceae which is widely distributed in trpical and warm temperature regins f the wrld (Wilsn et al. 2001). P. guajava cntains a number f phytcnstituents like tannins, essential il, flavnids, sapnins, lectins, vitamins, fiber, fatty acids, alanine, xalic acid, palmitic acid, quercetin, glutamic acid, d galactse and histidine (Li et al. 1999), Jaiarj 1999, Arima and Dann 2002, Begum et al. 2002, Gilani 2002, Michael et al. 2002, Jrdan et al. 2003). Plants are reservirs with healing pwers. A renewed interest in screening f plants fr bilgically active cmpunds, particularly the nes which effectively intercept the human ailments. Even tens f thusands f antimicrbial cmpunds exist, the ability f micrbes t develp resistance t even the mst pwerful antimicrbial cmpunds is amazingly rapid (Jayaraman et al. 2008). The mst ppular and attractive hypthesis is that vegetable and fruits cntain biactive cmpunds that have a prtective effect. Plant based extracts can be extracted frm any part f plant like seeds, rts, leaves, barks, fruits etc. One f such plant used traditinally is Psidium Received n February 25, 2012 Accepted n April 30, 2013 guajava Linn. The present wrk was carried ut t find pssible antimicrbial activities f the yung and mature leaves with respect t methanlic and aqueus extracts with reference t Gramnegative bacterial strains and a fungal strain using the agar cup diffusin methd, MIC and t crrelate the activity with the plant cnstituents present. MATERIALS AND METHODS The plant material fr the current studies was cllected frm the campus f VIVA Cllege, Virar (W) and which was authenticated (N ). The plant material was washed, dried at 40 C in ven fr a perid f seven days and then pwdered. Preparatin f Methanlic Extract Ten grams f dried pwdered material was first defatted with petrleum ether (6080 C) in Sxhlet apparatus fr 2 h. It was then extracted with methanl fr 6 h. The extract was filtered and the slvent was evaprated and recnstituted with DMSO (dimethyl sulfxide) fr further use. Of Aqueus Extract Water extract was prepared by adding ten grams f pwdered plant material with distilled water in a beaker n water bath with cnstant

2 ANTIMICROBIAL ACTIVITY OF LEAVES OF PSIDIUM GUAJAVA L. (GUAVA) 48 stirring, filtered, cncentrated. TEST ORAGANISM (NCIM 2803), (NCIM 2659) and (NCIM 3471) were used t check fr antimicrbial activity. The test rganisms were surced frm NCIM, Pune. Testing Fr Antimicrbial Activity The agar cup diffusin methd was used t check the antibacterial activity f the extract. 0.2mL f standardized bacterial stck 8 9) suspensins (10 10 clny frming units per ml was mixed with 20mL f sterile Mueller Hintn agar. The bulk seeded medium was pured int sterile Petri dishes and was left t set. Fr each testing rganism three such plates were prepared and in each f these plates 4 cups, 8 mm in diameter were cut using a sterile crk brer and the agar discs were remved. The cups were then filled with 40 µl extract f different cncentratins. Three replicates were carried ut fr each cncentratin f extract against each test rganism, simultaneusly additin f methanl and DMSO instead f extract were carried ut as cntrls. The plates were then incubated at 37 C fr 24 hr in upright psitin. After incubatin, the diameters f the results and grwth inhibitin znes were measured averaged and the mean values were tabulated. Minimum Inhibitry Cncentratin (MIC) The tube dilutin methd was used fr determinatin f minimum inhibitry cncentratin. The extract was serially diluted t give different cncentratin range (based n results btained fr Antimicrbial Susceptibility Testing (AST) fr each test rganism). A set f 9 sterile test tubes cntaining 5ml f sterile nutrient brth was prepared, 6 such tubes were inculated with 100 µl f bacterial culture suspensin and 1ml f plant extract and remaining tw tubes were inculated with 100 µl f bacterial culture, ne cntaining 1ml f DMSO and the ther cntaining Methanl. The last tube cntaining nly sterile nutrient brth was used as negative cntrl. The inculated tubes were then incubated at 37 C fr 24 hrs (fr bacterial cultures) and 25 C fr 2 days (fr fungal culture). After incubatin, the tubes were screened fr detecting the grwth which was cmpared against negative cntrl. The tube which did nt shw any grwth was taken as MIC f the extract. RESULTS AND DISCUSSION Antimicrbial Susceptibility Testing (AST) The methanlic and aqueus extract f Psidium guajava shwed antibacterial effect n bth Grampsitive and Gramnegative bacteria and als antifungal effect Antimicrbial activity was tested against Gram negative bacterium & and als against fungal strain. Minimum inhibitry cncentratin was als carried ut t find ut cncentratin f extract causing inhibitin f micrbial grwth. Methanlic extracts f bth yung and mature leaves shwed better results in cmparisn t aqueus extracts. MIC was calculated n the basis f the AST dne against the micrrganisms. P. shwed MIC best in methanlic extract f yung leaves, whereas E. and C. shwed MIC better in Methanlic extract f mature leaves. Aqueus extracts shwed MIC at higher cncentratin in cmparisn t methanlic extract (Table 1 and 2) As the plant matures physilgical changes takes place in different parts f the plant. The current study was carried ut t find the differences in the activity f leaves with different maturity. It was fund that methanlic extracts f bth the leaves gave gd results in cmparisn t aqueus extracts. P. shws mre susceptibility fr bth the extracts giving a zne measurement f 14mm and 13mm fr yung and mature leaves respectively, its MIC was fund t be 4.8% and 5 % fr methanlic extracts f yung & mature leaves respectively, 7 % and 6 % fr aqueus extracts f yung & mature leaves respectively, whereas E. shwed least susceptibility fr

3 MEENAKSHI VAIDYA 49 Table 1. Antimicrbial Susceptibilityactivity f Psidium guajava leaf extracts 1.Methanlic extract f yung leaves Organism Pseudmna s Organism Methanlic extracts f Psidium guajava(%) Mean inhibitin znes (mm) DMSO Methanl 13mm 13mm 12mm 12mm 11mm 11mm 13mm 12mm 12mm 11mm 12mm 11mm 11mm 10mm 10mm 2. Methanlic Extract f Mature leaves Methanlic extracts f Psidium guajava(%) Mean inhibitin znes (mm) ''Indicates n inhibitin (resistant t extract) DMSO Methanl 14mm 14mm 13mm 13mm 12mm 11mm 13mm 12mm 12mm 11mm 10mm 13mm 12mm 12mm 11mm 11mm 10mm 3. Aqueus Extract f Yung Leaves Organisms Aqueus extracts f Psidium guajava (%) Mean inhibitin znes (mm) Pseudmna s Organisms ''Indicates n inhibitin (resistant t extract) DMSO Methanl 12mm 12mm 11mm 11mm 11mm 11mm 12mm 11mm 11mm 4. Aqueus Extract f Mature leaves Aqueus extracts f Psidium guajava (%) Mean inhibitin znes (mm) ''Indicates n inhibitin (resistant t extract) DMSO Methanl 13mm 13mm 12mm 12mm 11mm 12mm 11mm 11mm 13mm 12mm 12mm 11mm ''Indicates n inhibitin (resistant t extract) Table 2.1 Minimum Inhibitry Cncentratin (Yung leaves) Methanlic extracts (%) Micrrganism DMSO Methanl Micrrganism DMSO Methanl Micrrganism DMSO Methanl +ve ve cntrl cntrl

4 ANTIMICROBIAL ACTIVITY OF LEAVES OF PSIDIUM GUAJAVA L. (GUAVA) Minimum Inhibitry Cncentratin (Mature leaves) Methanlic extracts (%) Micrrganism DMSO Methanl Micrrganism DMSO Methanl Cli Micrrganism DMSO Methanl +ve ve cntrl cntrl Minimum Inhibitry Cncentratin (yung leaves) Aqueus extracts (%) Micrrganism DMSO Methanl +v e c ntrl ve cntr l P seudmnas Micrrganism DMSO Methanl ve Micrrganism DMSO Methanl ve 2.4 Minimum Inhibitry Cncentratin (Mature leaves) Aqueus extracts (%) +ve ve Micrrganism DMSO Methanl cntrl cntrl Micrrganism DMSO Methanl ve Micrrganism DMSO Methanl ve aqueus extract f yung and mature leaves. Hwever, methanlic extracts shwed better results. The MIC results fr E. were 6.8% and 6% fr methanlic extract f yung and mature leaves and 9% & 8% fr aqueus extract. C. shwed results better than E. but lesser in cmparisn t P.. The results btained fr MIC were 6% and 5% fr methanlic extract f yung and mature leaves, and 8% and 7% fr aqueus extract f yung and mature leaves. Althugh there is nt a majr significance in the results. Fr mature and yung leaves still it can be said that the phytcnstituents present in yung and mature leaves d have a different mechanism and hence these plant materials shuld be screened prperly befre extractin f different phytcnstituents. REFERENCES Arima H & Dann G 2002 Islatin f antimicrbial cmpunds frm guava (Psidium guajava L.) and their structural elucidatin Bisci Bitechnl Bichem 66 (8)

5 MEENAKSHI VAIDYA 51 Begum S, Hassan S I & Siddiqui B S 2002 Tw new triterpenids frm the fresh leaves f Psidium guajava. Planta Med Gilani A H 2002 Triterpenids frm the leaves f Psidium guajava. Phytchemistry Jaiarj P 1999 Anticugh and antimicrbial activities f Psidium guajava leaves extract. J. Ethpharmacl Jayaraman K S, Manharan S M & IIachezian S 2008 Antibacterial, Antifungal and Tumr cell suppressin ptential f Mrinda Citriflia fruit extracts. Internatinal jurnal f integrative bilgy Jrdan M J, Margaria C A, Shaw P E & Gdner K I 2003 Vlatile cmpnents and arma active cmpunds in aqueus essence and fresh pink guava fruit puree (Psidium guajava L.) by GCMS and multidimensinal GC/GCO. J. Agric. Fd Chem Khan M L H & Ahmad J 1985 A pharmacgnstic study f Psidium guajava L., Internatinal Jurnal f Crude Drug Research Li J, Chen F & Lu J 1999 GCMS analysis f essential il frm the leaves f Psidium guajav.zhng Ya Cai Michael H N, Salib J Y & Ishak M S 2002 Acylated flavnl glycside frm Psidium guajava seeds. Pharmazie Wilsn P G, Brien O, Marcelle M, Gadek P A & Quinn C 2001 Myrtaceae Revised: A reassessment f Infrafamilial grups. Am. J. Bt

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