Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 183-196

ISSN: 2319-7706 Volume 3 Number 4 (2014) pp. 183-196 http://www.ijcmas.com

Original Research Article Preliminary phytochemical and in vitro control of selected pathogenic organisms by ethanolic extract of kola seeds

Obey Jackie1, T.Anthoney Swamy2* and Ngule Chrispus Mutuku2

1Department of Medical Laboratory Sciences, Faculty of MLS, University of Eastern Africa, Baraton, P.O. Box 2500, Eldoret, Kenya 2Department of Chemistry, University of Eastern Africa, Baraton, P.O. Box 2500, Eldoret, Kenya *Corresponding author

A B S T R A C T

Garcinia kola is cultivated for its economic importance. The is mainly used as a medicinal herb in West Africa. Although it is bitter the plant is used as a snack and a stimulant due to the high content of caffeine in the seeds of the plant. The current study was done to investigate the antibacterial activity of the plant seeds K e y w o r d s and to analyse the presence of important pharmaceutical compounds. From the study the plant seeds were found to contain tannins, saponins, flavonoids, Antibacterial terpenoids, glycosides and alkaloids but phenols and steroids were found to be activity; absent in the plant seeds. Among all the organisms tested with the ethanol extract Phytochemical; of G. kola, only B. cereus with an inhibition zone of 10.17±0.477 and E. coli with Kola; an inhibition zone of 12.83±0.833 were inhibited. All other organisms were not Medicinal inhibited. The penicillin control showed large zones of inhibition and DMSO did not show any zones of inhibition. The data obtained in this research is a scientific justification of the plants traditional use in the treatment of various stomach problems. From this research it is worthy to recommend the plant seeds for the treatment of diarrhea caused by E.coli and all the ailments caused by B. cereus, however further research needs to be done to isolate the pharmaceutical compounds, investigate their mode of action and the effect of the same in the in vivo environment.

Introduction

Traditionally plants are used as substitute to preservation of the knowledge, however drugs for various ailments affecting the trend is changing with many humankind. The information on medicinal communities abandoning there cultural value of plants conventionally was passed practices, this therefore creates the need from generation to generation. This for the documentation of the information passing of information some how has led o n traditional medicine in both the

183 Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 183-196 traditional way and also provide scientific A number of studies have been done to rationalization in order to increase the validate the use of traditional medicine in confidence on the use of plants as the treatment against microorganisms alternative means of treatment. Since time causing diseases. Studies have shown drug immemorial plants have been used as reaction and the side effects caused by novel source and reservoir of chemical synthetic drugs have increased the risk of agents with great restorative activities malignancy with fake and adulterated (Gobalakrishnan et al., 2013, Mensah et drugs increasing the problem of antibiotic al., 2009, Sindigawad, 2010). According resistance which on the other hand has to Panda et al., (2012) nature is a paradise imposed both biological and economic which offers medicinal principles to costs (Chabot et al., 1992., Chessin et al., humanity through plants. 1995 and Green, 2007). The failure of synthetic drugs to treat against various Plants consist of a wide spectrum of diseases has led to increased compounds which human beings can use hospitalization and mortality which have in dealing with the ailments which affect been associated with methicillin resistance them. When God created man, He put him Stapylococcus aureus (MRSA) infections in the Garden of Eden and then He planted (Arekemase et al., 2012). all kinds of beautiful trees. This marked the beginning of the human kind to use Medicinal plants have been tested plants as food. The creator had a purpose extensively and found to have great for providing human beings with plants pharmacological uses such as, anti- originally. Christian believe that it is sin inflammatory activity, antibacterial which brought sicknesses upon the world activity, anti- diabetic activity, anti-fungal and also hold to the believe that it is the activity, anti cancer activity, antioxidant continued disobedience to the creator activity, hepatoprotective activity, which continues to increase the ailments haemolytic activity, larvicidal activity, affecting humans and animals today. After anthelmintic activity, pain relief activity, man disobedient, God did not provide central nervous system activity, sexual alternative source of food to man and this impotence and erectile dysfunction and could impress that the first provision God hypolipidemic activity (Hosahally, et al, had given to the human kind was enough 2012, Anthoney et al., 2013., Farook et to sustain man. al., 2011., Kisangau et al., 2007., Nature provides us with great source of Kamatenesi-Mugisha et al., 2005., Adu et cheap and safe alternative drugs. al., 2011., Deep et al., 2007., Joshi et al., According Ellen G. White (1897), the 2011., Arivoli et al., 2012., Ngule et al., Creator of the universe has given some 2014., Anthoney et al., 2014). simple herbs of the field that at times are beneficial and if every society was Garcinia kola tree is cultivated for its educated on the use of these medicinal economic importance. The plant is used as herbs in case of sickness, such suffering a medicinal plant in West Africa. might be prevented, and no doctor would Although it is bitter the plant is used as a be required. These old-fashioned, simple snack and a stimulant due to the high herbs, used intelligently, would have content of caffeine in the seeds of the recovered many sick people, who have plant. The plant is used in folk medicine to died under drug medication. treat against gram positive bacteria, Ebola

184 Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 183-196 virus infections, Flu, dysentery and vacuum evaporator (R -11) with a water diarrhea (Iwu, 1993). According to bath at 40oC. The extract was brought to Chinyere et al. (2013), the plants bark, dryness using vacuum and pressure pump seeds and stem are traditionally used in the at room temperature. The residue was then treatment of throat infections, acute fever obtained and used for the experiment. and inflammation of the respiratory tract. The leaves are also used to treat against Qualitative phytochemical analysis stomach problems and also a remedy for anthelmintic and typhoid (Irrine, 1981., The phytoconstituents analysis of extracts and Gill, 1992). The current study was for identification of bioactive chemicals done to investigate the antibacterial was done using standard procedures activity of the plant seeds and to analyse (Trease and Evans, 1989., Harbone, 1973 the presence of important pharmaceutical and Sofowara, 1993). compounds. 1. Tannins Materials and Methods About 0.5 g of the sample was put in a test Sample collection and preparation tube and 20 ml of distilled water was added and heated to boiling. The mixture The herb Garcinia kola was bought from was then filtered and 1 % of FeCl3 was Monrovia, in the West Africa added to the filtrate and observations region. The samples were identified by a made. A brown green color or a blue black taxonomist in the University of Eastern coloration indicates the presence of Africa, Baraton. The samples were cut in tannins. to small pieces and spread to dry at room temperature in the chemistry laboratory for 2. Saponins about three weeks. They were then ground into fine powder and put in transparent The crude solvent extract was mixed with polythene bags. 5 ml of water and vigorously shaken. The formation of stable foam indicates the Extraction procedure presence of saponins.

Using electric analytical beam balance 3. Flavonoids fifty grams of the powdered seeds of the Garcinia kola were placed in 500 ml A portion of the aqueous extract was conical flask, ethanol was added until the added in a test tube. To this, 5 ml of dilute samples were completely submerged in the ammonia and 2 ml of concentrated solvent. The mixture was then agitated for sulphuric acid were added. The thorough mixing. The mixture was kept appearance of a yellow color indicated the for 24 hours on a shaker for effective presence of flavonoids. extraction of the plant components. The extract was filtered using Butchner funnel; 4. Terpenoids Whatman number 1 filter paper and a vacuum-pressure pump. The filtrate was The extracts of the plant material was re-filtered again using the same apparatus. taken in to a clean test tube 2 ml of The solvent was evaporated using rotary chloroform was added and vigorously

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Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 183-196 shaken and then evaporated to dryness. To Bioassay study the residue, 2 ml of concentrated sulphuric acid was added and heated for Preparation of the Bacterial Suspension about 2 minutes. A grey color indicates the presence of terpenoids. The turbidity of each of the bacterial 5. Glycosides suspension was prepared to match to a 0.5 McFarland standard. The McFarland Salkowsks test standard was prepared by dissolving 0.05 g of BaCl2 in 50 ml of water to obtain a The extract of the plant material was 1% solution of Barium chloride (w/v). mixed with 2 ml of chloroform and 2 ml of This was mixed with 99.5 ml of 1% concentrated sulphuric acid was carefully sulfuric acid solution. Three five added and shaken gently, then the identical colonies of each bacterium was observations were made. A red brown taken from a blood agar plate (Himedia) color indicate the presence of steroidal culture and dropped in Mueller Hinton ring (glycone portion of glycoside). broth (Himedia). The broth culture was incubated at 370C for 2 - 6 hours until it 6. Alkaloids achieved turbidity similar to the 0.5 McFarland standards. The culture that The crude extract was mixed with 1% of exceeded the 0.5 McFarland standard were HCl in a test tube. The test tube was then each adjusted with the aid of a UV heated gently and filtered .To the filtrate a spectrophotometer to 0.132A0 at a few drops of Mayer s and Wagner s wavelength of 600 nm in order to obtain reagents were added by the side of the test an approximate cell density of 1x108 tube. A resulting precipitate indicates the CFU/ml. presence of alkaloids. Preparation of the Extract 7. Steroids Concentrations and Antibiotic

Libermann Burchard reaction Stock solutions for the extract were prepared by dissolving 500 mg in 1 ml of About 2 g of the extract was put in a test dimethylsulfoxide (DMSO). An tube and 10 ml of chloroform added and antibiotic control was made by dissolving filtered. Then 2 ml of the filtrate was 1µg of penicillin in 1 ml of sterile distilled mixed with 2 ml of a mixture of acetic water. DMSO served as a negative acid and concentrated sulphuric acid. Blue control. green ring indicate the presence of steroids. Determination of bioactivity of the Extract 8. Phenols Mueller Hinton agar plates were prepared The plants extract was put in a test tube as per the manufacturer s instruction. The and treated with a few drops of 2% of plate containing the medium were FeCl3 blue green or black coloration inoculated and sterile paper disc indicate the presence of phenols. (whatmann no. 1, 5mm diameter) were

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Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 183-196 impregnated with the 500mg/ml solution tannins can be used to treat hereditary of the extract (100µl) and allowed to dry. hemochromatosis which is a hereditary Three discs containing the plant extract disease characterized by excessive and two discs one containing penicillin as absorption of dietary iron. According to the positive control and the other Chung et al. (1998), many tannin containing DMSO as the negative were molecules have shown the ability to then spaced on the surface of the sterile reduce the mutagenic activity of a number petri dishes using sterile pair of forceps. of mutagens. The anti-carcinogenic and The discs were labeled on the underside of anti-mutagenic potentials of tannins may the plate. The plates were incubated at be related to their antioxidative property 370C for between 24 to 48 hours and the which is important in protecting cellular zones of inhibition were measured in oxidative damage including lipid millimeters with the aid of a transparent peroxidation. The growths of many fungi, ruler. yeast, bacteria and viruses have been proven to be inhibited by tannins. Tannins Results and Discussion have also been reported to exert physiological effects, such as to accelerate From the study the extract of Garcinia blood pressure, decrease the serum lipid kola was found to contain tannins, level, and produce liver necrosis and saponins, flavonoids, terpenoids, module immune responses. The dosage glycosides, alkaloids but steroids and and kind of tannins are critical to these phenols were found to be absent (Table. effects (Chung et al., 1998). 1). Tannins are secondary metabolites which were found to be present in the The presence of Saponins shows the plant seeds. They are glycosides of gallic potential of the plants to be used to or protocatechvic acids. There astringent produce mild detergents and intracellular property makes them useful in preventing histochemistry staining to allow antibody diarrhea and controlling hemorrhage due access to intercellular proteins (Maobe et to their ability to precipitate proteins, al., 2013). They have been found to treat mucus and constrict blood vessels hypercholesterolemia, hyperglycemia, (Kokwaro, 2009). This is the reason why antioxidant, anti-inflammatory, central traditional healers use plants reach in nervous system activities, anticancer and tannins to treat wounds and burns since weight loss (Maobe et al., 2013). They are they are able to cause blood clotting. used to stop bleeding, treating wounds and Some tannins have been reported to inhibit ulcers as it helps red blood cells to HIV replication selectively besides the use precipitate and coagulate (Just et al., of diuretics (Argal and Pathak, 2006). 1998). This can be attributed to the ability This shows how traditional plants rich in of saponins to bind with glucose and tannins can be used to control this cholesterol molecules. Saponins have also dangerous disease. Tannins have also been associated with inhibitory effect on shown antiparasitic effects (Akiyama et inflammation (Just et al., 1998).Saponins al., 2001). According to Bajal (1988), are used by the folkloric remedies of tannins can also be used to protect the Kashmir (India) in treating wounds (Foster kidney since when taken the poliovirus, and Duke, 1990) this is because of their herpes complex virus and various enteric ability to cause red blood cells coagulation viruses are inactivated. Foods rich in and therefore help in blood clotting,

187 Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 183-196 treating wounds and enteric ulcers property which is useful in improving problems (Chiej, 1984). Saponins have blood circulation in brain and in also been used to prevent Alzheimer disease (Sharma, 2006). Leaf hypercholesterolemia, antibiotic activity, extract of Ginkgo biloba which contains anti-inflammatory and anti-diabetic. flavonoids was used for improving blood circulation in brain varix. Several isoflavone can be used to improve blood Flavonoids are secondary metabolites with circulation. Free radicals including the polyphenolic structure and synthesized in hydroxyl, hydrogen peroxide, superoxide plants, through polypropanoid and lipid peroxide have been associated pathway(Ghasemzadeh et al., 2011). with a number of diseases such as Flavonoids have been classified in to six cardiovascular disease, cataracts, diabetes, sub-groups which include flavones, gastrointestinal inflammatory diseases, flavanol, flavanone, flava-3-ols, isoflavone cancer, asthma, liver disease, macular and anthocynidin. Flavonoids are known degeneration, periodental disease and to contain specific compounds called other inflammatory processes. These antioxidants which protect human, animal oxidants are produced during normal body and plant cells against the damaging chemical processes. They can be damaged effects of free radicals. Imbalance between through free-radical damage. free radicals and antioxidants leads to oxidative stress which has been associated Terpenoids have medicinal value such as with inflammation, autoimmune diseases, ant-carcinogenic, antimalarial, cataract, cancer, Parkinson s disease, antimicrobial and diuretics activity aging and arteriosclerosis. It also plays a (Deganhardt, 2003 & Pichersky and role in heart diseases and Gershezon, 2002). Evaluation of the ant- neurodegenerative diseases. Flavonoids inflammatory activity of three different are used as antioxidants because of their Copaiba oleoresins showed that the crude ability to scavage free radicals such as extract of the plant and its fractions of peroxide and hydroperoxide of lipid hexane, dichloromethane and methanolic hydroxyl hence inhibiting oxidation that extracts of other plant such as C. lead to degenerative diseases (Samatha et cearensis, C. reticulata and C. multijuga al., 2012). They can be used as anti- have anti-inflammatory potential (Viega et diabetic. According to Namki (1990), al., 2007). Terpenoids have also shown a flavonoids can be used to prevent great potential in treatment against disease synthesis of flavors that are caused by fat causing microorganisms. Terpenoids have oxidation. Flavonoids have been found to exhibited antibacterial activity against E. have antibacterial activity due to their coli, Staphylococcus, Pseudomonas ability to complex with extracellular and aeruginosa (Piera et al., 2011), Proteus soluble proteins and to complex with mirabilis (Piera et al 2011), Klebsiella bacterial cell wall (Marjorie, 1999). pneumoniae (Piera et al., 2011 & Santo et Flavonoids are produced by plant in al., 2008), methillin-resistant S. aureus, response to microbial infection and studies Staphylococcus epidermidis (Santo et al., have shown that they have antibacterial 2008), Listeria monocytogenes (Nero et activity against a wide range of micro- al., 2010), Enterobacter cloacae, yeast; organisms (Yadav and Agarwala, 2011). Candida albicans and fungi, Aspergillus Flavonoids have also vasodilator activity a flavus (Leandro et al., 2012). Terpenoids

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Table.1 Phytochemical analysis results

Phytochemical Observation Inference Tannins Brown black color Present Saponins Stable foam Present Flavonoids Yellow color Present Terpenoids Black-grey Present Glycosides Green brown Present Alkaloids Precipitate Present Steroids No blue green ring Absent Phenols No black coloration Absent Table.2 Antimicrobial Activity of Garcinia kola (bitter kola) against selected pathogenic microorganisms Extract Penicillin DMSO Microorganisms mean ±S.E (mm) mean ±S.E (mm) mean ±S.E. (mm) Serratia marcescens 0.00±0.000 32.33±0.760 0.00±0.000 Bacillus cereus 10.17±0.477 29.00±0.516 0.00±0.000 Escherichia coli 12.83±0.833 21.50±0.563 0.00±0.000 Proteus vulgaris 0.00±0.000 25.83±0.307 0.00±0.000 Salmonella sp 0.000±0.000 28.83±0.703 0.00±0.000 Key: S.E. = Standard error Table.3 Turkey s honestly significant Difference among micro - organisms using 500mglml of Garcinia kola (bitter kola) extract Comparison P-value Significance S. marcescens vs B. cereus 0.000 S S. marcescens vs E.coli 0.000 S S. marcescens vs P. Vulgaris 1.000 NS S. marcescens vs Salmonella s.p 1.000 NS S. marcescens vs S. marcescens control 0.000 S B. cereus vs E. coli 0.020 S B. cereus vs P. vulgaris 0.000 S B. cereus vs Salmonella s.p 0.000 S Salmonella s.p. vs P. vulgaris 0.000 S B. cereus vs B. cereus control 0.000 S E. coli vs P. vulgaris 0.000 S E. coli vs Salmonella s.p. 0.000 S E. coli vs p. Vulgaris 0.000 S E.coli vs E.coli control 0.000 S P. Vulgaris vs Salmonella s.p. 1.000 NS P. Vulgaris vs P. Vulgaris control 0.000 S Salmonella s.p. vs Salmonella s.p. control 0.000 S

189 Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 183-196 have also been found to reduce the growth 2004). Cryptolepine has also been used of melanoma cells on mice after oral clinically to treat malaria, colic and administration (Lima et al., 2003). stomach ulcers (Boye and Ampufo, 1983), Terpenoids have been proved scientifically and also used in anticancer drugs. to kill mosquito larvae. Terpenoids According to Karou et al. (2006), much extracted from C.reticulata species have study has been done on pharmacological shown potential in killing A. aegypti properties of alkaloids and proved to have (Geris et al., 2003). antiprotozoal, cytotoxic and anti- inflammatory properties. Glycosides are secondary metabolites from plants or animal sources in which a The presence of alkaloids in the plant sugar is bound to a non-carbohydrate justifies its medicinal value. Alkaloids moiety. The term Glycoside is a collective have been isolated from different plants term used for compounds formed with a and their medicinal values tested. The glycosidic bonding between a sugar and most important use of alkaloids already another compound other than sugar. known with its originality from plants is Cardiac glycosides have been used the use of alkaloids compounds in the traditionally as arrow poisons or as heart treatment of malaria. According to drugs. They are used to strengthen the Ameyawn and Duker-Eshon ( 2009), heart and make it function properly under many of the antimalarial drugs used today controlled therapeutic dose. Cardiac are quinoline derivatives manipulated glycosides bind to and inhibit Na+/K+- from cinchona species bark (Garnham, ATPase, inhibition of Na+/K+-ATPase 1966). Alkaloids have been identified for raises the level of sodium ions in cardiac their functions which include analgesic, myocytes, which leads to an increase in antiplasmodic and antibacterial activity the level of calcium ions and an increase (Okwu and Josiah, 2006). According to in cardiac contraction force (Schatzmann Ayitey and Addae (1977), bitter leaves and Rass., 1965). The unexpected results containing alkaloids are capable of relating glycosides with anticancer reducing headache associated with properties have created a great interest in hypertension. this secondary metabolite. This has lead to clinical trial of glycosides based drugs in Among all the organisms tested with the clinics (Newman et al., 2008). methanol extract of G. kola, only B. cereus and E. coli were inhibited (Table 2). All Alkaloids which are secondary other organisms were not inhibited. The metabolites, they can be defined as a penicillin control showed large zones of cyclic compounds which have nitrogen in inhibition and DMSO did not show any a negative oxidation state. They affect the zones of inhibition. Analysis of variance chemical transmitters action of the showed there was significant differences nervous system. They also have other in the zones of inhibition among the pharmacological activities such as bacterial organisms tested (p<0.001). analgesic, antispasmodic, antihypertensive A multiple comparison with the Tukey s effects and anti-arrhymic effects and test (Table.3) showed that the zones of antibacterial effect. Cryptolepine a major inhibition of S. marcescens was alkaloid in the plant S.acuta was found to significantly lower than that of B. cereus, be an antimalarial agent (Banzouzi et al., lower than E. coli and S. marcenscens

190 Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 183-196 control (p<0.001). There was no Streptococcus pyogenes and 37.0mm for significant difference between S a l m o n e l la typhi, (Ezeanya and Daniel, S. marcenscens and P. vulgaris and 2 0 1 3 ) . Between 5mg/ml to 25mg/ml of S. marcenscens and Salmonella sp. Garcinia kola and Cola nitida showed (p>0.05). B. cereus had significantly zones of inhibition against E.coli, S. higher zones of inhibition than all the aureus, S. typhi and K. pneumoniae. The organisms (p<0.001) but had significantly methanol soluble extract of G. kola lower zones of inhibition than E. coli showed zones of inhibition of 20mm, (p<0.05). E. coli also had significantly 25mm, 24mm, 9mm and 20mm against E. higher zones of inhibition than P. vulgaris coli, S. aureus, S. typhi and K. pneumonia and Salmonella sp. (p<0.001). There was respectively. That of Cola nitida showed no significant difference in the zones of 23mm, 9mm, 18.5mm, 12.5mm against E. inhibition of P. vulgaris and Salmonella coli, S. aureus, S.typhi, K pneumonia sp. (p>0.05). All the penicillin controls respectively. This study showed that Cola zones of inhibition against each organism nitida is a better antimicrobial against E. were significantly higher than those for the coli while G. kola is good for S. aureus, S. treatment with the extracts against the typhi and K. pneumonia, (Indabawa and organisms. Arzai, 2011).

The study is in conformity with other From the data obtained in this study it is studies in which Polyisoprenyl therefore worthy to mention that the plant benzophenone, kolanone in the petroleum can be used to treat against all the ether and hydroxyl biflavanols in the ethyl infections caused by Bacillus cereus and acetate fraction of G. kola showed activity even other species of Bacillus viz self- against gram positive and gram negative limited gastroenteritis, post traumatic bacteria (bacteriostatic) and against wounds, surgical wounds infections, Candida albicans and Aspergillus flavus burns, ocular infections such as (fungistatic). MIC of the GB 1 fraction endophthalmitis, corneal abscess and was 3.1x 10-7mg/ml and 3.0x10-3mg/ml panophthalmitis (Sankararaman and for E. coli (Madubunyi, 1995). In an agar Velayuthan, 2013 & Garcia-Arribas et al., well diffusion assay, G. Kola was tested 1988). The plant compounds can also be against several microorganisms and used to treat immunologically showed that the ethanol extracts exhibited compromised patients including AIDS and zones of inhibition ranging from 17 to 23 malignant disease victims (Cotton et al., mm while the aqueous hot water extract 1987 & Tuazon et al., 1979). The plant s showed zones ranging from 20 to 27 mm. capability to inhibit the growth of E. coli is Antifungal activity was shown against a scientific prove that the plant can be Staphylococcus aureus at 0.008mg/ml. used to a great extent to treat against Phytochemical compounds such as enteric infections caused by the bacteria. flavonoids, tannins saponins, steroids, cardiac glycosides and reducing sugar The presence of the important where found to be present (Arekemaje et phytochemicals is in conformity with the al., 2012). The acetic acid extract of results obtained in previous studies in Garcinia kola has also shown zones of which the plant seeds medicinal value was inhibition as high as 17.5mm for S. attributed to the presence of flavonoids, aureus, 18.5mm for E.coli, 35.0mm for tannins, saponins, triterpenes,

191 Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 183-196 benzophenones and xanthones (Iwu, 1982 Oono, T., Iwatsuki, K. 2001. and Geiger, 1988). The medicinal value of Antibacterial action of several tannins the plant could directly be attributted to against Staphylococcus aureus. J. one or a combination of two or more Antimicrobe. phytochemicals found in the plant to give Ameyaw, Y. and Duker-Eshun, G. a synergistic effect on the ailment been 2009.The alkaloid content of the treated against. The antibacterial activity ethano-plant organs of three observed in this study could be directly antimalarial medicinal plants species attributed to the presence of these in the eastern region of . important phytochemicals as indicated in Int.J.Chem, 7(1):48-58. previous studies (Ngule et al., 2013 and Anthoney, S.T., & Ngule, C.M. 2013. Anthoney et al., 2013). Chemical constituents of infused Plectranthus argentatus leaves. World The data obtained in this research is a Journal of Publisher, 1(3): 151-160. scientific justification the plants traditional Anthoney, S.T., Jackie, O., Ngule, C.M. use in the treatment of various stomach 2013. In vitro control of selected problems. From this research it is worthy pathogenic organisms by Vernonia to recommend the plant seeds for the adoensis roots. International Journal treatment of diarrhea caused by E.coli and of Pharmacy and Life Sciences, 4(8): all the ailments caused by B. cereus, 2855-2859. however further research needs to be done Anthoney, S.T., Ngule, C.M. & Obey, J. to isolate the pharmaceutical compounds, 2013. In vitro control of selected investigate their mode of action and the pathogenic organisms by Vernonia effect of the same in the in vivo adoensis. Int. J. of Pharm. & Sci, 4(8): environment. 2855-2859. Anthoney, S.T., Ngule, C.M. and Acknowledgment Machoka, R. 2014. Histopathological effects of Vernonia adoensis roots The authors of this paper are very much extract on the larvae of the mosquito thankful to the Department of Chemistry Culex quinquefanciatus. Journal of and Medical Laboratory Science, Pharmaceutical Biology, 4(1): 1-3. University of Eastern Africa, Baraton. Anthoney, S.T., Ngule, C.M. and Obey, J. Authors also thank to taxonomist Mr. Joel 2013. Phytochemical analysis of Ochieng Ondiek, University of Eastern Vernonia adoensis leaves and roots Africa, Baraton for identifying the plant. used as a traditional medicinal plant in Kenya. International Journal of References Pharmacy and Biological Sciences, 3(3): 46-52. Adu, F., Gbedema, S.Y., Akanwariwiak, Anthoney, S.T., Ngule, C.M. and W.G., Annan, K., and Boamah, V.E. Obey.2013. Phytopharmacological 2011. The effects of Acanthospermum analysis of methanolic aqua extract hispidum extract on the antibacterial (fractions) of Senna didymobotrya activity of amoxillin and ciprofloxacin, roots. International Journal of Journal for drugs and Medicines, 3(1): Bioassays, 02(11): 1473-1479. 58-63. Arekemaje, M.O., Aliyu., M.B., Kayode, Akiyama, H., Fujii, K., Yamasaki, O., R.M.O., Ajiboye A. E., Ajijolakewu

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