ANTIMICROBIAL ACTIVITY OF LEAF EXTRACTS OF CARICA
Research Articleejbps, 2018, Volume 5, Issue 7, 323-327.SJIF Impact Factor 4.918ISSN 2349-8870European ofJournalof Biomedical and Pharmaceutical SciencesEuropean JournalBiomedicalVolume: 5Issue: 7AND Pharmaceutical sciencesSaket et al.323-327Year: 2018http://www.ejbps.comANTIMICROBIAL ACTIVITY OF LEAF EXTRACTS OF CARICA PAPAYA AGAINSTINFECTION-CAUSING BACTERIA1Abhilasha Shrivastava, 2Bhajan lal Saket, 2Bharat Choudhary1Professor, Dept. of Botany, Govt. Model Science College, Rewa (M.P.)Research Scholar, Centre for Biotechnology Study, A.P.S.University, Rewa (M.P.)2*Corresponding Author: Bhajan Lal SaketResearch Scholar, Centre for Biotechnology Study, A.P.S. University, Rewa (M.P.).Article Received on 28/04/2018Article Revised on 18/05/2018Article Accepted on 08/06/2018ABSTRACTDue to increase in the thrust for the production of plant-based antimicrobials, the present study was performed onCarica papaya leaves. The study revealed that papaya leaves could contain active antimicrobial compounds whichmay hinder the growth of wound infection-causing pathogens in invitro conditions The leaf extract was preparedby using acetone, methanol, and water. The antimicrobial nature of the extract was studied by agar well diffusionmethod against wound infection-causing pathogens viz., Escherichia coli, Staphylococcus aureus, Proteusvulgaris, Klebsiella pneumoniae, and Pseudomonas aeruginosa. The acetone leaf extracts exerted pronouncedantibacterial effect on gram negative bacteria especially Pseudomonas sp. The results obviously justified theimportance of topical application of papaya leaf extracts to treat the wound infection as a traditional practice.KEYWORDS: Plant-based antimicrobials, Pseudomonas sp., acetone, methanol, and water.1. INTRODUCTIONEmergence of resistant strains of pathogenicmicroorganism has also continued to pose a major healthconcern about the efficacy of several drugs, mostimportantly antibiotics in current use (Timothy and Idu,2011). The search for newer sources of antibiotics is aglobal challenge pre-occupying research institutions,pharmaceutical companies, and academia, since manyinfectious agents are becoming resistant to syntheticdrugs (Latha and Kannabiran, 2006).The local use of natural plants as primary healthremedies, due to their pharmacological properties, isquite common in Asia, Latin America, and Africa,(Bibithaet al., 2002).Sofowora (1982) and Balandrin et al. (1985) definedmedicinal plants as a plant in which one or more organscontain substances that can be used for therapeuticpurposes or which it precursors for the manufacturing ofdrugs are useful for disease therapy. Since medicinalplants do not nearly save people from feeling pain butpermit them to emerge unscathed, they deserveinvestigation.Each part of papaya tree possess economic value when itis grown on a commercial scale (Krishna et al., 2008).Even though the active components are normallyextracted from all parts of the plant, the concentration ofthese components vary from structure to structure.www.ejbps.comPapaya (Carica papaya Linn) is commonly known for itsfood and nutritional values throughout the world. Themedicinal properties of papaya fruit and other parts ofthe plant are also well known in traditional system ofmedicine. However, parts known to contain the highestconcentration of the principles are preferred fortherapeutic purposes and it can either be the leaves, stem,barks, roots, bulks, corms, rhizomes, woods, flowers,fruits, and the seeds (Kafaru, 1994). C. papaya belongsto the family Caricaceae. It is known by a variety ofnames, viz., pawpaw, papaya, papayer, pepol, tinti, chichput, fan kua, wan shou kuo, Kavunagaci, kepaya etc.Various parts of the papaya plant, which include theleaves, fruit, seed, latex, and root, are known to containbioactive compounds. The plant parts are found topossess some properties like analgesic, amebicide,antibacterial, cardiotonic, cholagogue, digestive,emenagogue, febrifuge, hypotensive, laxative, pectoral,stomachic, and vermifuge (Afolayan, 2003).The leaves of the papaya plants contain chemicalcompounds of karpain, substance which killsmicroorganisms that often interfere with the digestivefunction (Udoh et al., 2005). Chymenopapain and papainare the two important bioactive compounds present in C.papaya. Papaya leaves are made as tea for the treatmentof malaria. Antimalarial and anti-plasmodial activity hasbeen noted in some preparations of the plant. Papayaleaf-extracts have phenolic compounds, such asprotocatechuic acid, p-coumaric acid, 5, 7-323
Saket et al.European Journal of Biomedical and Pharmaceutical Sciencesdimethoxycoumarin, caffeic acid, kaempferol, quercetin,and chlorogenic acid (Romasi et al., 2011; Peter et al.,2014). During the last few decades, considerableprogress has been achieved regarding the therapeuticproperties of papaya.2.2. Pathogen Used in the Present StudyThe test pathogens were procured in Centre forBiotechnology Studies, A.P.S. University and then theywere further reconfirmed by morphological, cultural andBiochemical characteristics. The cultures wereemulsified in 5ml of Nutrient Broth (NA) and incubatedfor 24 hrs. Fresh cultures were employed for assessingantibacterial activity of the papaya leaf-extracts.The use of C. papaya L (Caricaceae) in traditionalmedicine relies on papain, the active principle whichexerts an ulcer protective effect. The C. papaya possessantimicrobial, antioxidant, and anti-inflammatoryactivities. It is reported to heal chronic ulcer. ShivanandaNayak et al. (2007) examined the wound healing activityof the leaf extract of C. papaya in experimentallyinduced excision and dead space wounds in diabetic rats.The wound healing processes are further worsened bythe entry of pathogens. It is common traditional practiceto treat the wound with the leaf-extract of papaya toaccelerate the healing action. There are many reportsavailable which demonstrate the wound-healing propertyof the papaya leaves. In addition, papaya leaves possessantibacterial activity which might prevent themultiplication of wound infection-causing bacteria. Withthis in mind, the present investigation was undertaken topredict the antibacterial properties of papaya leavesagainst some wound-infection causing pathogens and tojustify plant-based compounds could replace syntheticones.2.3. Crude Extract PreparationThe crude extract from the leaves of papaya wasprepared according to the method proposed by Alabi etal. (2012). The aqueous extract was prepared bysuspending 100g of powdered leaves in 200ml ofdistilled water. This mixture was diluted with 300ml ofdistilled water, and then allowed to stand for 24 hrs. Theresulting extract was decanted and filtered through aWhatman filter paper. The filtrate was then concentratedwith rotary evaporator at 45ºC. (this is the sameprocedure adopted for preparing acetone and methanolextracts – 100g of powder 500ml of 95% ethanol, 100gof powder 500ml of 95% acetone)2.4. Test for Antibacterial ActivityAgar well diffusion method was adopted to assess theantibacterial activity of papaya leaf-extracts againstwound pathogens. The crude extracts were furtherdiluted with diluent at the concentration ranging25mg/ml, 50mg/ml, 75mg/ml, and 100mg/ml. For thetest, Muller-Hinton agar plates were swabbed with testorganism, and the wells (5mm) were filled with differentconcentration of the extracts. After incubation for 24 hrs,the antibacterial efficiency of the leaf-extract weredetermined by measuring the zone of inhibition formedaround the well.2. MATERIALS AND METHODS2.1. Collection of LeavesDisease free, fresh, young, and green leaves werecollected from the papaya plants. The leaves werewashed thoroughly 3–10 times in sterile distilled water.Then, they were air-dried under shade at roomtemperature for 8 days and finely powdered using ablender.2.5. RESULTInvestigation on Antibacterial Activity of Carica Papaya Leaf Extracts against Wound Infection-CausingBacteriaTable 1: Antibacterial activity of acetone extract of Carica papaya leaf on wound infection causing pathogens(well diffusion method).Diameter of zone of inhibition (mm)S.NO Wound pathogens12345www.ejbps.comStaphylococus aureusEscherichia coliKlebsiella pneumoniaProteus vulgarisPseudomonas 12111317mg/ml1213141524324
Saket et al.European Journal of Biomedical and Pharmaceutical SciencesTable 2: Antibacterial activity of methanol extract of Carica papaya leaf on wound infection causingpathogens (well diffusion method).Diameter of zone of inhibition (mm)S.NO Wound us aureus0010142345Escherichia coliKlebsiella pneumoniaProteus vulgarisPseudomonas aeruginosa0000121011131312142417151626Table 3: Antibacterial activity of aqueous extract of Carica papaya leaf on wound infection causingpathogens (well diffusion method).Diameter of zone of inhibition( mm)255075100S.NO Wound pathogensmg/mlmg/mlmg/mlmg/ml1Staphylococus aureus000102Escherichia coli0011113Klebsiella pneumonia0010114Proteus vulgaris0012135Pseudomonas aeruginosa0014153. RESULT AND DISCUSSIONThe antimicrobial properties of plants have beeninvestigated by a number of studies worldwide and manyof them have been used as therapeutic alternativesbecause of their antimicrobial properties (Adriana et al.,2007). Plants are the cheaper and safer alternativesources of antimicrobials (Pretorius et al., 2001;Doughari et al., 2007).Anibijuwon and Udeze (2009) extracted bioactivecompounds from leaf and root of C. papaya using waterand organic solvents, which were investigated forantibacterial activity against some human pathogenicbacteria. Both leaf and root extracts showed pronouncedinhibition against gram positive bacteria than the gramnegative bacteria tested, but the highest activity was seenwith Pseudomonas aeruginosa. In our present study, thebetter inhibition was seen with acetone extract againstPseudomonas aeruginosa than the other organisms tested(Table 1).Ocloo et al. (2012) studied the efficacies of crudeextracts of C. papaya seeds against Staphylococcusaureus, Escherichia coli and Shigella flexneri using discdiffusion method. The crude organic (acetone, methanol)extracts inhibited the growth of all three organisms. Theaqueous extract of C. papaya leaves were investigatedfor evaluation of wound healing potential in rats. Theseresults strongly document the beneficial effects of plantextract for the acceleration of wound healing process inrats (Mahmood et al., 2005).In our present investigation, all the extracts preparedfrom leaf (acetone, water, and methanol) exhibitedwww.ejbps.comhighest antibacterial activity against gram negativeorganism. These results are in agreement with resultsdeclared by many researches. Nirosha andMangalanayaki (2013) also reported gram negativebacteria are more susceptible to the extracts of papayaleaf and stem. But the results of Suresh et al. (2008)showed the antibacterial activity of the papaya leafextract was more pronounced on gram positive thangram negative bacteria. According to Jigna and Sumitra(2006), the plant leaf-extracts are more active againstgram positive than gram negative bacteria. Ourinvestigation results are contrary with their findings.Ogunjobi and Elizabeth (2011) reported that leaf andseed extract of C. papaya have inhibitory effect nas aeruginosa, Pseudomonas fluroescens,and Salmonella typhi. They also demonstrated thatethanol extracts contributed much more antibacterialactivity than aqueous extract (Ezeifeka et al., 2004). Ourfindings are correlated with their results, herein, acetoneextracts demonstrated the highest antibacterial activitythan aqueous and methanol extracts (Tables 1–3). Thehighest antibacterial activity brought about by organicsolvent might be due to better solubility of the activecomponents of leaves.From our present study, it could be concluded that thepapaya leaves might effectively inhibit the growth ofcertain wound infection-causing pathogens without anyside effects. The active principle compound may beextracted and further purified, and incorporated as a basecompound for the preparation of topical ointment.325
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