Antibacterial activity of Ocimum gratissimum AND Gongronema latifolium ON Staphylococcus aureus AND Salmonella typhi

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1 Antiteril tivity of Oimum grtissimum AND Gongronem ltifolium ON Stphyloous ureus AND Slmonell typhi Bnkole, H. A. 1, Anjorin, A. A. 2*, Kzeem, M. I. 1, Ogehe, M. E. 1, Agfor, U. 1 1 Deprtment of Biohemistry, Lgos Stte University, Ojo, Lgos, Nigeri 2 Deprtment of Miroiology, Lgos Stte University, Ojo, Lgos, Nigeri *Corresponding Author: Emil: zlortories@yhoo.o.uk; dul-zeez.njorin@lsu.edu.ng Astrt Oimum grtissimum nd Gongronem ltifolium hve een used y herl mediine prtitioners in omintion with other hers in the tretment of Stphylool nd Slmonelll infetions. The im of this study ws to evlute ntiteril tivity of extrts of Oimum grtissimum (Efinrin) nd Gongronem ltifolium (Utzi) on Stphyloous ureus nd Slmonell speies. Extrts were otined y queous nd ethnoli methods with onentrtion rnge of 25% to 75%. The dimeter of inhiition zones y queous extrt of Oimum grtissimum ws etween 7-15 mm while tht of the ethnoli extrt ws etween 7 nd 12 mm. The minimum inhiitory onentrtion (MIC) ws 25% for the queous extrt of O. grtissimum on S ureus with no effet on Slmonell, while tht of the ethnoli extrt ws 75% for oth orgnisms. G. ltifolium showed MICs of 25% nd 75% for the queous extrt on S. ureus nd Slmonell respetively while the ethnoli extrts showed MICs of 25% nd 50% on S. ureus nd Slmonell respetively. Minimum teriidl onentrtion (MBC) for the queous extrts of G. ltifolium were 50% nd 75% for S ureus nd Slmonell respetively while tht of the ethnoli extrt ws 25% for S ureus nd 75% for Slmonell. O. grtissimum showed MBC t 75% onentrtion only for the queous extrt lone on S ureus. Therefore, the results otined indited tht the extrts of O. grtissimum nd G. ltifolium possess ntiteril tivities ginst S. ureus nd Slmonell speies. Key words: Antiteril tivity, Oimum grtissimum, Gongronem ltifolium, Stphyloous ureus, Slmonell typhi 114 ISSN (Online version) ejsiene@gmil.om

2 Introdution Spies hve long history of oth ulinry nd mediinl uses (Tpsell nd Swrhi, 2006). Some spies hve een reported to hve oth teriidl nd teriostti tivities (Onwuliri nd Wonng, 2005). They re generlly reognized to e sfe, either euse of their trditionl use without ny doumented detrimentl impt or euse of dedited toxiologil studies (Smid nd Gorris, 1999). Some spies re highly mediinl nd hve een used to sustin helth nd ure illnesses (Osunwole, 1999). Some of the ftors tht determine their ntimiroil tivities inlude onentrtion nd omposition of the spies, the type of miroorgnism, ph vlue, temperture nd phenoli sustnes present in the food (Sgdi, 2003). Oimum grtissimum elongs to the fmily Leguminoee ommonly known s Alfv. It is found in tropil nd wrm temperte regions suh s Indi nd Nigeri, prtiulrly in the svnnh nd ostl res (Okigo nd Ogonny, 2006). It is generlly sserted to possess vrious ulinry nd mediinl properties. The mediinl properties exert oth teriostti nd teriidl effets on some teri (Okigo nd Igwe, 2007). It hs een used in the tretment of different diseses suh s upper respirtory trt infetion, dirrhoe, hedhe, onjuntivitis, skin disese, pneumoni, tooth nd gum disorders, fever nd s mosquito repellents (Onjoi, 1986; Ilori et l., 1996; Okigo nd Mnek, 2008) nd tive ginst severl speies of teri inluding Listeri monoytogenes, Shigell, Slmonell nd Proteus (Nwosu nd Okfor, 1995; Akinyemi et l., 2005). Gongronem ltifolium, known s utzi in the south estern prt of Nigeri is tropil rinforest plnt whih elongs to the fmily Asepidee (Ugohukwu nd Bdy, 2002; Akinnug et l., 2011). It hs ntiteril tivity ginst Stphyloous ureus nd Esherihi oli nd shows strong urtive properties. It is thus used for the mngement of vrious diseses used y these orgnisms (Akinyemi et l., 2005). Slmonell is fulttive neroe tht uses foodorne disese tht result to ondition known s Slmonellosis while Stphyloous ureus on the other hnd hs een implited in severl illnesses nd stomh upset (Mosset nd Vn, 1990). Stphyloous ureus produes het stle toxin whih is mjor puli helth thret, eing one of the ommon use of hospitl nd ommunity quired 115 ISSN (Online version) ejsiene@gmil.om

3 infetions (Aires-de-Sour et l., 2006). This study ws undertken therefore, to investigte the ntiteril properties of the rude ethnoli nd queous extrts of Oimum grtissimum (efinrin) nd Gongronem ltifolium (utzi) ginst food-orne pthogens of Stphyloous ureus nd Slmonell Speies. Mterils nd methods Smple olletion The plnts used in this study, Oimum grtissimum (Efinrin) nd Gongronem ltifolium (Utzi) were purhsed from the vegetle setion of Bdgry lol mrket in Bdgry Lol Government Are, Lgos Stte, Nigeri. The plnts were txonomilly identified t the Deprtment of Botny, Lgos Stte University, Ojo. The plnt prts used re the stlk nd leves. Growth hit of the plnts Oimum grtissimum is n nnul her whih grows in severl regions ll over the world (Sjjdi, 2006, Simon et l., 1990). It grows in the ool moist nd tropil rin forest zones in nnul tempertures etween 6-24 C nd reeiving mm nnul preipittion (Gryer et l., 1996). Gongronem ltifolium (Aslpidee) is perennil edile plnt with soft nd plile stem. It is hrvested from forest in southestern sttes of Nigeri nd some other prts of Su-Shrn Afri (Okfor, 1995). Miroorgnisms The test miroorgnisms, Stphyloous ureus nd Slmonell speies were otined from the Deprtment of Miroiology, Lgos Stte University, Ojo, Nigeri. All the ultures were otined in their pure form. Preprtion of plnt extrts The fresh spies were seprted, lened nd wshed in sterile distilled wter, dried, weighed nd powdered finely using eletri lender. After otining fine powder, the spies were weighed. 25g of ir dried mteril ws shken in 250ml of 96% (w/v) ethnol t room temperture for the ethnoli extrtion nd in 250ml of ssterile distilled wter for the queous extrtion. The mixtures were llowed to stnd for 24 hours fter whih they were filtered using fine mesh loth. The solvents were evported to dryness t temperture of 40 0 C using wter th nd the extrts were then reonstituted in sterile distilled wter to mke onentrtions of 25%, 50% nd 75%. 116 ISSN (Online version) ejsiene@gmil.om

4 Antiteril sensitivity testing using filter pper method Filter pper diss of 6mm were prepred from Whtmn No.1 filter pper nd sterilized. Using ethnol dipped nd flmed foreps, the diss were inserted into the vrious onentrtions of the extrts nd pled septilly over the nutrient gr nd slmonell-shigell gr pltes seeded with the test miroorgnisms. A totl of five diss were pled on eh plte, with three for the vrious spie onentrtions, one for hlormpheniol whih served s positive ontrol nd the lst dis for distilled wter whih served s the negtive ontrol. The inoulted pltes were inuted t room temperture for 24 hours. The ntiteril tivity ws evluted y mesuring the zones of inhiition, whih is the ler zone round the vrious diss in millimetres. Dilution method for inhiitory effet nd teriidl effet (qulittive test) Ten millilitre of eh spie extrt ws inoulted with 0.1ml of pthogeni miroil ulture nd mixed well. Two hundred miroliters of this mixture ws pipetted into the mirorry pltes. After inution t 37 0 C for 24 hours, the wells were heked for turidity, spots, dots or pellets formed t the se of the wells. Any growth t prtiulr onentrtion represented minimum inhiitory onentrtion. Smples from the mirorry pltes tht did not show ny growth or turidity were used to inoulte fresh sterile roth tht ontined no spie extrts. 100ul of this ulture ws reintrodued into fresh 100ul nutrient roth. The lowest onentrtion of the spie extrt tht yielded no turidity or spots or dots of growth following this seond inoultion or su-ulturing showed the minimum teriidl onentrtion. Sterile distilled wter inoulted with teril strins without spies ws used s ontrol. Results Tle 1 shows the minimum inhiitory onentrtion (MIC) of the queous nd ethnoli extrt of Gongronem ltifolium nd Oimum grtissimum ginst S. ureus nd Slmonell typhi. At ll the onentrtions tested, queous nd ethnoli extrts of G. ltifolium inhiited the growth of S. ureus. However, it only inhiited the growth of S. typhi t high onentrtions. Aqueous extrt O. grtissimum inhiited the growth of S. ureus t 50 nd 75% onentrtion while it only inhiited S. typhi t 75% onentrtion. 117 ISSN (Online version) ejsiene@gmil.om

5 The Minimum Bteriidl onentrtion (MBC) of queous nd ethnoli extrt of Gongronem ltifolium nd Oimum grtissimum ginst S. ureus nd Slmonell typhi is depited in tle 2. Aqueous nd ethnoli extrts of G. ltifolium inhiited S. ureus t ll onentrtions exept 25% queous extrt while O. grtissimum extrts hd no effet exept t 75% queous extrt. As for S. typhi, extrts of G. ltifolium only inhiited the miro-orgnism t the highest onentrtion while O. grtissimum hd no effet. The ntiteril tivities of queous extrt of G. ltifolium with respet to the inhiition dimeters (mm) on the test orgnisms re shown in figure 1. The inhiition dimeter of S. ureus t 25 nd 50% re not signifintly different (p 0.05) from eh other. However, the inhiition dimeters t ll the onentrtions of the extrt were signifintly different from the ontrol (p 0.05). At ll the onentrtions tested, the ethnoli extrt of G. ltifolium were signifintly different (p 0.05) from one nother nd tht of the ontrol (figure 2). Figures 3 nd 4 showed the ntiteril tivities of queous nd ethnoli extrts of O. grtissimum on oth S. ureus nd S. typhi. At ll the onentrtions tested, the inhiition dimeters of the extrts were signifintly different (p 0.05) from one nother nd to the ontrol. However t 25% onentrtion, queous extrt of O. grtissimum hd no effet on S. typhi (figure 3). 118 ISSN (Online version) ejsiene@gmil.om

6 Tle 1: Minimum inhiitory onentrtion (MIC) of queous nd ethnoli extrt of Gongronem ltifolium nd Oimum grtissimum ginst S. ureus nd Slmonell typhi Smples Conentrtion S. ureus Slmonell typhi G. ltifolium 25% ++ - Aqueous Extrt 50% % Control - - G. ltifolium 25% ++ - Ethnoli Extrt 50% % Control ++ - O. grtissimum 25% - - Aqueous Extrt 50% % ++ - Control - - O. grtissimum 25% - - Ethnoli Extrt 50% % Control No growth inhiition; ++ Complete growth inhiition 119 ISSN (Online version) ejsiene@gmil.om

7 Tle 2: Minimum Bteriidl onentrtion (MBC) of queous nd ethnoli extrt of Gongronem ltifolium nd Oimum grtissimum ginst S ureus nd Slmonell typhi Smples Conentrtion S ureus Slmonell G. ltifolium 25% - - Aqueous Extrt 50% % Control - - G. ltifolium 25% ++ - Ethnoli Extrt 50% % Control - - O. grtissimum 25% - - Aqueous Extrt 50% % ++ - Control - - O. grtissimum 25% - - Ethnoli Extrt 50% % - - Control No growth inhiition; ++ Complete growth inhiition. 120 ISSN (Online version) ejsiene@gmil.om

8 Inhiition dimeters(mm) Inhiition dimeters(mm) Est Afrin Journl of Siene nd Tehnology, 2012; 2(1): % Extrt 50% Extrt 75% Extrt Chlormpheniol Distilled wter 0 S.ureus Slmonell typhi Figure 1: Antiteril tivity of queous extrt of Gongronem ltifolium. Groups with the sme letters re not signifintly different t (p < 0.05) d d % Extrt 50% Extrt 75% Extrt Chlormpheniol Distilled wter 5 0 S.ureus Slmonell typhi Figure 2: Antiteril tivity of ethnol extrt of Gongronem ltifolium. Groups with the sme letters re not signifintly different t (p < 0.05) 121 ISSN (Online version) ejsiene@gmil.om

9 Inhiition dimeters(mm) Inhiition dimeters(mm) Est Afrin Journl of Siene nd Tehnology, 2012; 2(1): d % Extrt 50% Extrt 75% Extrt Chlormpheniol Distilled wter 0 S.ureus Slmonell typhi Figure 3: Antiteril tivity of queous extrt of Oimum grtissimum. Groups with the sme letters re not signifintly different t (p < 0.05) d d % Extrt 50% Extrt 75% Extrt Chlormpheniol Distilled wter 0 S.ureus Slmonell typhi Figure 4: Antiteril tivity of ethnol extrt of Oimum grtissimum. Groups with the sme letters re not signifintly different t (p < 0.05) 122 ISSN (Online version) ejsiene@gmil.om

10 DISCUSSION Antimiroil tivities of queous nd ethnoli extrts of O. grtissimum nd G. ltifolium were exmined for the presene or sene of teril growth, zone of inhiition nd turidity. signifint differenes (p < 0.05) etween the vrious onentrtions. This finding shows ethnol to e etter solvent thn wter in the extrtion of the tive priniple of this plnt. Okigo nd Ogonny (2006) stted tht the oserved The results showed tht the extrts hve onentrtion dependent inhiitory effet on the test orgnisms. The queous extrt differenes etween the plnt extrts my e due to insoluility of tive ompounds in wter s in the se of G. ltifolium or of Oimum grtissimum showed higher the presene of inhiitors to the tivity thn the ethnoli frtion. This ntimiroil omponents. finding justifies the ethno- medil use of O. grtissimum leves s plster to over These results suggest tht the wound surfes nd y s ord fter it is soked in wter. This study lso supports the pplition of O. grtissimum in dermtologil rems ginst wound onentrtion of O. grtissimum ws not suffiient for the test miroorgnisms unlike the G. ltifolium whih showed MBC vlues for oth the queous nd infetion used y S. ureus in ethnoli frtions on oth orgnisms. ordne with the work of Pphn et l., (1994). Although, muh reserh hs not een rried out on the ntimiroil tivity of G. ltifolium whih hs een used for ges y the people of West Afri prtiulrly Nigerins for dietry nd mediinl purposes (Ej et l., 2011), ntimiroil tivity reveled in this study grees with the work of Nwinyi et l., (2009) who reported the ntimiroil This intivity my e due to ge of plnt, extrting solvent, method of extrtion nd time of hrvesting of plnt mterils nd lso tht vritions my lso e due to the different tive sustnes present in these plnts nd the ility of the different tive sustnes to effetively extrt these sustnes (Amdoih nd Oi, 1999; Okigo nd Ajle, 2005; Okigo nd Ogonny, 2006). tivities of G. ltifolium ginst S. ureus. The ethnoli extrts of G. ltifolium however were found to e more Previous report reveled tht O. effetive thn the queous extrt with grtissimum is rih in phytohemils like 123 ISSN (Online version) ejsiene@gmil.om

11 lkloids, tnnins, glyosides, steroidl terpenes nd flvonoids (Nweze et l., 2004). Flvonoids hve een reported to hve ntioxidnt tivity nd re effetive svengers of superoxide nions thus they n signifintly ffet the ell wll of the miroorgnisms whih my invrily led to the ollpse of the ell wll nd overll, ffet the entire mehnism of the miroil ell (Nwinyi et l., 2009). G. ltifolium hs een reported to onsist of through their nturl inhiitory mehnisms, either y inhiiting or killing the pthogens. This study hs provided the sis for the use of these two plnts in the tretment of ilments used y S. ureus nd Slmonell typhi. Sine the solvents used hd vrying effets on the tive priniples of these plnts, further studies need to e done in determining the est strtegy to e dopted in their extrtion nd dministrtion. flvonoids, tnnins, sponins, polyphenols, lkloids nd hydrogen ynide (Atnghwo et l., 2009). The presene of tnnins, lkloids, flvonoids, sponins nd polyphenols my e responsile for the ntiteril tivity of G. ltifolium (Irhim et l., 2006; Andy et l., 2008; Ej et l., 2011). Fromi (2003) noted tht the tive omponents of some Nigerin mediinl plnts reside in the phytohemil onstituents mentioned ove. Wht hs not een resolved is the seprtion of the speifi iotive omponents ginst speifi orgnisms nd this hs een noted to ffet qulity nd sfety of herl mediines (Ej et l., 2011). CONCLUSION This study showed tht the two plnts, efinrin nd utzi possess vrying degrees REFERENCES Aires-de-Sous, M., Boye, K., de Lenstre, H., Deplno, A., Enright, M. C., Etienne, J., Friedrih, A., Hrmsen, D., Holmes, A., Huijsdens, X. W., Kerns, A. M., Mellmnn, A., Meugnier, H., Rsheed, J. K., Splurg, E., Strommenger, B., Struelens, M. J., Tenover, F. C., Thoms, J., Vogel, U., Westh, H., Xu, J., nd Witte, W., (2006). High Inter lortory Reproduiility of DNA Sequene-Bsed Typing of Bteri in Multienter Study. Journl of linil miroiology, 44(2), Akinnug, A. M., Bmidele, O., Ekehi, P. nd Adeniyi, O. S., (2011). Effets of n Ethnoli Lef Extrt of Gongronem ltifolium on Some Hemtologil of ntimiroil tivity. These spies t 124 ISSN (Online version) ejsiene@gmil.om

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15 Legends to figures Figure 1: Antimiroil tivity of the queous extrts of G ltifolium ginst S. ureus nd Slmonell typhi t onentrtions of 25%, 50% nd 75% with hlormpheniol nd distilled wter serving s positive nd negtive ontrols respetively. Figure 2: Antimiroil tivity of the ethnoli extrts of G ltifolium ginst S ureus nd Slmonell typhi t onentrtions of 25%, 50% nd 75% with hlormpheniol nd distilled wter serving s positive nd negtive ontrols respetively. Figure 3: Antimiroil tivity of the queous extrts of O. grtissimum ginst S. ureus nd Slmonell typhi t onentrtions of 25%, 50% nd 75% with hlormpheniol nd distilled wter serving s positive nd negtive ontrols respetively. Figure 4: Antimiroil tivity of the ethnoli extrts of O grtissimum ginst S. ureus nd Slmonell typhi t onentrtions of 25%, 50% nd 75% with hlormpheniol nd distilled wter serving s positive nd negtive ontrols respetively. 128 ISSN (Online version) ejsiene@gmil.om