Bioactivity and Phytochemical Screening of Nigerian Medicinal Plants Growing in Ondo and Ekiti State Against Bacterial Isolates from Pediatrics Hospital

Journal of Advances in Medical and Pharmaceutical Sciences 4(2): 1-14, 2015, Article no.JAMPS.18805 ISSN: 2394-1111

SCIENCEDOMAIN international www.sciencedomain.org

Bioactivity and Phytochemical Screening of Nigerian Medicinal Plants Growing in Ondo and Ekiti State against Bacterial Isolates from Pediatrics Hospital

Osuntokun Oludare Temitope1*

1Department of Microbiology, Faculty of Science, Adekunle Ajasin University, Akungba Akoko, P.M.B 001, Ondo State, Nigeria.

Author’s contribution

Author OOT is the sole author designed, analyzed, interpreted and prepared the manuscript for publication. He also designed the materials and methods used in the course of the research work and also prepare the first draft of the manuscript and the literature search. Author OOT read and approved the final manuscript for the final publication. Author OOT is a researcher and has researched into the antimicrobial and phytochemical properties of various medicinal plants in Nigerian and Africa.

Article Information

DOI: 10.9734/JAMPS/2015/18805 Editor(s): (1) Dongdong Wang, Department of Pharmacogonosy, West China College of Pharmacy, Sichuan University, China. (2) Palmiro Poltronieri, National Research Council of Italy, CNR-ISPA, Italy and Institute of Sciences of Food Productions, Operative Unit in Lecce, Italy. Reviewers: (1) Anonymous, National Autonomous University of Mexico, México. (2) Anonymous, University of Maringá, Brazil. (3) Anonymous, Manonmaniam Sundaranar University, India. (4) Thomas Eidenberger, Dept. of Bio- and Environmental Technology, Univ. of Applied Sciences Upper Austria, Austria. Complete Peer review History: http://sciencedomain.org/review-history/10321

Received 11th May 2015 th Original Research Article Accepted 4 July 2015 Published 25th July 2015

ABSTRACT

The Aqueous, ethanol, hexane and ethyl acetate extracts of the leaves, bark and roots of Acacia albida, Anchomanes difformis, Boscia senegalensis, Bridelia ferruginea, Ficus ingens, Indigofera arrecta and Moringa oleifera were tested for possible source of antibacterial activities against selected clinical organisms found in the pediatrics hospital in Ondo and Ekiti State, Nigeria. The selected clinical organisms used for this research work are Salmonella typhi, Staphylococcus aureus, Klebsiella pneumoniae and Escherichia coli. It was observed that all plant were potent against the clinical test organism. Anchomanes difformis, Moringa oleifera and Indigofera arrecta ______

*Corresponding author: E-mail: [email protected];

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extracts were the most active medicinal plants during the course of this research work. Boscia senegalensis, Acacia albida, Bridelia ferruginea and Ficus ingens extract were not as active against the test organism. All the medicinal plants extract shows evidence of antibacterial properties. Phytochemical screening of the medicinal plants reveals, very important and vital phytochemicals like saponin, tanins, alkaloids, flavonoids, cardiac glucosides and anthraquinones, at a very appreciable quantity.it also reveals presence of minerals like sodium, potassium, calcium, iron, zinc, copper and phosphorus at mg/100 g which are essential to biological and physiological activities. The main objectives of this research work are to investigate the antimicrobial potency and phytochemical activities of seven Nigerian Medicinal plants on Selected Clinical Organisms.

Keywords: Antibacterial assay; agar well diffusion method; phytochemical screening; minerals composition; nutrient and anti-nutrient composition; proximate composition.

1. INTRODUCTION The medicinal plants is taking a place of pride in the world health economy, knowing fully well The discovery of new antibiotics is a very that, developing countries can make earn a lot important aspect of medicinal plants, which is foreign exchange in this aspect of drug dated as far back as 500 to 700 BC. Pharmacist, production, drug discovery, development of new Pharmacologist, Biochemist and Pharmaceutical drugs and antibiotic through medicinal plants, microbiologist are taking the necessary clue from especially in Nigeria, where God has blessed us the advent of medicinal plants in the discovery abundantly with various degree of medicinal and production of new antibiotic for the cure of plants [7,8]. different diseases [1]. Traditional medicine like orthodox medicine has its own methods and techniques of applications, Medicinal Plants are good source of antimicrobial which however aims at finding a cure to various agents, which can be used as a form of health hazards [9]. The treatment and control of chemotherapy in the cure of different illness. diseases by the use of the available medicinal Some are used as chewing sticks, some are plants in a locality will continue to play a used for soup condiment and some Medicinal significant role in medical health care. Nearly all plants are use even for spiritual purpose in the cultures and civilizations from ancient times to South Western part of Nigeria [20]. Major health the present day have depended fully or partially hazard may not be cure by the conventional on herbal medicine because of their medicine, but medicinal plants are found to be effectiveness, affordability, availability, low useful in such areas. Poor people may not even toxicity and acceptability. Our empirical belief in have enough money to buy the conventional Nigeria and Africa on herbal drugs gives us an drug, but nature has blessed us with the edge on the use of medicinal plants which is on medicinal plants for food and also for the cure of the high increase in our daily activities, due to major health hazard [2,3]. ineffectiveness of most conventional drugs as a result of microbial resistance in some health The use of medicinal plants for therapy challenge in the developing countries, more predates the introduction of synthetic patients in Nigerian are seen in herbal medical antibiotics. It also predates the religious, social centers than ever. The intractable problem of and economic barriers [4]. Antibiotics are antimicrobial resistance has led to the naturally occurring or synthetic organic resurgence of interest in herbal products as compounds which inhibit or destroy bacteria, sources of novel compounds to suppress or generally at low concentration [5]. Many lives possibly eradicate the ever increasing problems have been saved by the use of synthetic of emergence of newer diseases [10,11]. antibiotics but in recent times, these disease causing microbes have developed resistance to Phytochemicals are plants produce chemical some of these antibiotics and have become compounds as part of their normal metabolic difficult to treat in our various health care activities. These phytochemicals are divided into homes,an example of our pediatric hospital. This two, this are the primary metabolites such as situation may lead to increased cost of sugars and fats found in all plants; and medication and in the long run leads to increased secondary metabolites which are compounds morbidity and mortality [6]. found in a smaller range of plants, serving a

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more specific function. It is these secondary The plant parts were authenticated by a Botanist metabolites and pigments that can have at the Department of Botany, Ekiti State therapeutic actions in humans and which can be University Ado Ekiti, and Adekunle Ajasin refined to produce drugs examples are inulin University, Akungba Akoko, Ondo State ,Nigeria. from the roots of dahlias, quinine from the A voucher specimen number 1784 was assigned cinchona, morphine, codeine and digoxin from to it for future reference. the foxglove [12] Plant profile, local and botanical names and its Alkaloids are a class of chemical compounds uses, are elucidate in the Table 1, to show that containing a nitrogen ring. Alkaloids are this plant are been in use for centuries [3,7]. produced by a large variety of organisms, including bacteria, fungi, plants, and animals, 2.2 Source of Microorganisms and are part of the group of natural products (also called secondary metabolites). Many Pure cultures of Salmonella typhi, alkaloids can be purified from crude extracts by Staphylococcus aureus, Klebsiella pneumoniae, acid-base extraction. Examples are the local Escherichia coli isolated from children stools anesthetic and stimulant cocaine; the attending the Pediatrics from the Federal Medical psychedelic psilocin; the stimulant caffeine; Center (Pediatrics) (FMC), Owo, Ondo State, nicotine; the analgesic morphine; the and Ekiti State Teaching Hospital (Pediatrics), antibacterial berberine; the anticancer compound Ekiti State, Nigeria Bacterial cultures were vincristine; the anti-hypertension agent reserpine; maintained on Blood agar slant. Authenticities of the choline-mimeric galatamine; the spasmolysis the isolates were preform through the use of API agent atropine; the vasodilator vincamine; the kits 20 [6]. anti-arhythmia compound quinidine; the anti- asthma therapeutic ephedrine; and the 2.3 Extract Preparation antimalarial drug quinine [13,14,15]. The root and branch of the test plant were well

dried and then grinded to powder. About 200 g In view of this, it is therefore very important to of the powder were separately soaked in 400 mL search for effective, efficient, low cost and of 95% ethanol, distilled water and ethyl acetate reliable traditional therapeutic agents, for the and Hexane in a 500 mL reagent bottle and development of locally made drugs (antibiotics), stoppered. This was allowed to stand for 14 days to reduce abuse of drugs and drug resistant to permit full extraction of the active ingredients. organisms which is a common factors in some The fluids were then filtered using Whatman No1 conventional drugs [16]. The main objectives of filter paper. The extracts were rotary dried to this research work is to investigate the obtain the concentrate. It was then kept in fridge antimicrobial potency and phytochemical at- 4ºC prior to use [6]. activities of seven Nigerian Medicinal plants on selected clinical organisms. 2.4 Antimicrobial Assay (Determination of Zones of Inhibition) Using Agar 2. MATERIALS AND METHODS Well Diffusion Method

The experiment involved seven types of Nigerian Twenty five milliliters of Mueller-Hinton agar medicinal plants, with four different extracting were poured into sterile petri dishes and allowed solvent, four clinical organisms, and three to set. Standardized test bacterial cultures were concentrations of medicinal plants extracts, the inoculated into the sterile Kliger iron agar plate four clinical isolates are Bacteria. using sterile cotton swab. A sterile cork borer of 6mm diameter were used to punch wells on 2.1 Source of Plant Materials the agar on each of the petri dishes. Three holes were made on the surface of the plate with The Nigerian Medicinal plants selected were one in the center to serve as the control. Each obtained from the roots and the branch of Acacia hole was labeled (A,B,C,D) representing a albida, Anchomaens difformis. Boscia particular concentration of 20 mg/mL, 40 mg/mL senegalenss, Bridelia ferruginea, Ficus ingens, and 60 mg/mL. The dishes were then filled into Indigofera arrecta and Moringa oleifera. The the wells with three drop (2 mL) of their plant were collected in Ikare Akoko, a tropical respective type of extract according to the rainforest, Ondo State, Nigeria with latitude labeled format. The central well containing the (7.21692 North) and longitutide (5.21561 East) diluent reagent Streptomycin was used as

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control. The process was carried out for each precipitates indicate the presence of Alkaloids extract and the inoculated petri dishes were [16]. left for few minutes for extract of diffuse into agar. The plates were incubated at 37ºC for 18- 2.6.2 Test for tannins 24 hours after which the zones of inhibition (if any) were measured [17,18,19]. One milliliter of the filtrate was mixed with 2 mL of FeC1, A dark green colour indicated a positive 2.5 Phytochemical Screening test for the tannins.

The extract were analysed for the presence of Alkaloid, Glycosides, Tannins, Saponins, 2.6.3 Test for saponins Anthraquinones, Anthocyanosides, Flavonoids, Reducing sugars, Cyanogenic [14,18,20,3]. One milliliter of the plant filtrate were diluted with 2 mL of distilled water; the mixture were 2.6 Qualitative Method of Analyses vigorously shaken and left to stand for 10 minutes during which time, the development of Plant filtrates were prepared by boiling 20 g of foam on the surface of the mixture lasting for the fresh plant in distilled water. The solution was more than 10 mm, indicates the presence of filtered through a vacuum pump. The filtrates saponins. were used for the phytochemical screening for flavonoids, tannins, saponins, alkaloids, reducing 2.6.4 Test for anthraquinones sugars, anthraquinones and anthocyanosides.

2.6.1 Test for alkaloids One milliliter of the plant filtrate was shaken with 10 mL of benzene; the mixture was filtered and 5 About 0.2 g was warmed with 2% of H2SO4 for mL of 10% (v/v) ammonia were added, then two minutes, it was filtered and few drops of shaken and observed. A pinkish solution Dragendoff's reagent were added. Orange red indicates a positive test.

Table 1. Profile of the medicinal plants under study

Botanical name Family Local names Part used Medicinal uses 1. Acacia albid. a Leguminosae Gawo Root Skin infections 2. Anchomanes difformis. Araceae Chakara Roots 1. Cough. 2. Respiratory diseases. 3. Boscia senegalensis. Capparidaceae Anza Roots 1. Anticancer. 2. Ulcer swellings. 4. Bridelia ferruginea. Euphorbiaceae Kirni Leaf 1. Antibiotic. 2. Skin infection, 3. Diuretic, 4. Febrifuge urethral discharge. 5. Ficus ingens. Moraceae Kawuri Root 1. Piles. 2. Diarrhea. 3. Laxative. 4. Diuretic. 6. Indigofera arrecta. Papilionoideae Ba-ba Leaf 1. Infections stomach. 2. Pain. 3.Snakebite. 4.Epilepsy. 7.Moringa oleifera. Moringacceae Zogale Leaf 1Asthma. 2.Eye infection. 3.Migraine. 4.Headache. 5.Febrifuge. 6.Abortifacient. Sofowora(2008) [2], Osuntokun (2014) [3]

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2.6.5 Test for anthocyanosides were added. The combined n-butanol extracts were washed twice with 10 mL of 5% aqueous One milliliter of the plant filtrate was mixed with sodium chloride. The remaining solution was 5mL of dilute HCI; a pale pink colour indicates heated in a water bath. After evaporation, the the positive test. samples were dried in the oven to a constant weight, the saponin content was calculated as 2.6.6 Test for flavonoids percentage of the starting material [6].

One milliliter of plant filtrate were mixed with 2 2.7.2 Flavonoids mL of 10% lead acetate; a brownish precipitate indicated a positive test for the phenolic About 10 g of the plant sample were extracted flavonoids. While for flavonoids, I mL of the plant repeatedly with 100 mL of 80% aqueous filtrate were mixed with 2 mL of dilute NaOH; a methanol, at room temperature. The whole golden yellow colour indicated the presence of solution was filtered through Whatman filter flavonoids. paper No 42. The filtrate were later transferred into a crucible and evaporated into dryness over 2.6.7 Test for reducing sugars a water bath; the dry content was weighed to a constant weight [14]. One milliliter of the plant filtrate was mixed with Fehling A and Fehling B separately; a brown 2.7.3 Cardiac glucosides colour with Fehling B and a green colour with Fehling A indicate the presence of reducing Legal tests (the killer-kiliani) were adopted as sugars. follows to detect the presence of Cardiac Glucosides. 0.5 g of the extract was added to 2 2.6.8 Test for cyanogenic glucosides mL of acetic anhydrate plus H2S04 [16].

This was carried out subjecting 0.5 g of the 2.7.4 Tannins extract 10 mL sterile water filtering and adding sodium picrate to the filtrate and heated to boil About 500 mg of the plant sample were weighed [16]. into a 50 mL plastic bottle. 50 mL of distilled water was added and shaken for 1 hour on a 2.6.9 Test for cardiac glucosides mechanical shaker. This was filtered into a 50 mL volumetric flask and made up to the marked Legal test (the killer-kiliani) were adopted as level. Then, 5 mL of the filtrate was transferred follows, 0.5 g of the extract were added to 2 mL into a test tube and mixed with 2 mL of 0.1 M of acetic anhydrate plus H2S04 [16]. FeCl in 0.1 M Hydrochloride and 0.008 M Potassium ferrocyanide. The absorbance was 2.7 Quantitative Method of Analyses measured at 120 nm within 10 minutes. The tannins content was calculated [16]. 2.7.1 Saponins 2.7.5 Alkaloids About 20 g each of dried plant samples were ground and, put into a conical flask after which 5 g of the plant sample were weighed into a 250 100 mL of 20% aqueous ethanol were added. mL beaker and 200 mL of 10% acetic acid in The mixture was heated using a hot water bath. ethanol was then be added, the reaction mixture At about 55ºC, for 4 hours with continuous were covered and allowed to stand for 4 hours. stirring, after which the mixture were filtered and This was filtered and the extract will be the residue re-extracted with a further 200 mL of concentrated on a water bath to one-quarter of 20% ethanol. The combined extracts were the original volume. Concentrated ammonium reduced to 40 mL over a water bath at about hydroxide was added drop-wise to the extract 90ºC.The concentrate was transferred into a 250 until the precipitation is complete. The whole mL separatory funnel and 20 rnL of diethyl ether solution were allowed to settle and the precipitate were added and then shaken vigorously. The was collected, washed with dilute ammonium aqueous layers were recovered while the ether hydroxide and then filtered; the residue being the layer was discarded. The purification process alkaloid, which was dried and weighed to a was repeated three times. 60 rnL of n-butanol constant mass [16].

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2.7.6 Phlobatannins 2.8.5 Determination of ash value

About 0.5 g of each plant extracts were dissolved The ash values were obtained by heating in distilled water and filtered. The filtrates were samples at 550ºC in a muffle furnace (Wise boiled in 2% HCl, red precipitate show the Therm, FHP-03, Korea) for 3 hours [24]. present of phlobatannins. 2.8.6 Determination of carbohydrate content 2.7.6.1 Total phenol (Spectrophotometric methods) The carbohydrate content was determined by

2 g each of the samples were defatted with 1mL subtracting the total crude protein, crude fiber, of diethyl ether using a soxhlet apparatus for 2 ash content and crude fat from the total dry hours. The fat free samples were boiled with 50 matter [26]. mL of ether for the extraction of the phenolic components for 15 minutes. 5 mL of the extracts 2.8.7 Determination of fiber content crude were pipetted into 5 mL flask and then 10 mL distilled water was added. 2 mL of ammonium Fiber was estimated by acid-base digestion with hydroxide solution and 5 mL of concentrated 1.25% H2SO4 (v/v) and 1.25% NaOH (w/v) amyl alcohol were also added. The samples solutions [25]. were made up to mark and left to react for 30 minutes. For color development. This was 3. RESULTS measured at 505 nm [6,21,22]. Table 2 shows the Antibacterial Activity of 2.8 Determination of Proximate Analysis Aqueous Extract of Medicinal Plants at 20 of Medicinal Plants mg/mL, 40 mg/mL, and 60 mg/mL Concentration. The medicinal plants used for this bioassay are The proximate parameters (moisture, dry matter, Acacia albida, Anchomanes difformis, Boscia ash, crude fats, proteins and fibers, nitrogen, senegalensis, Bridelia ferruginea, Ficus ingens, carbohydrates and energy values) were Indigofera arrecta, Moringa olefera and the determined using Association of Official clinical organism are Salmonella typhi Analytical Chemists Methods. Staphylococcus aureus, Klebsiella pneumoniae , Escherichia coli. The control for the experiment 2.8.1 Determination of moisture is Streptomycin at 20 mg/mL concentration. determination of moisture

Content was done by drying samples in oven Table 3 shows the Antibacterial Activity of (Wise Ven, WON-50, Korea) at 110ºC until Ethanol Extract of Medicinal Plants at 20 mg/mL, constant weight was attained [23]. 40 mg/mL, and 60 mg/mL Concentration. The medicinal plants used for this bioassay are 2.8.2 Determination of nitrogen content Acacia albida, Anchomanes difformis, Boscia senegalensis, Bridelia ferruginea, Ficus ingens, Nitrogen estimation was carried out by the micro- Indigofera arrecta, Moringa olefera and the Kjeldahl (BUCHI, KjelFlex K-360, Switzerland) clinical organism are Salmonella typhi method with some modification [24]. Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli. The control for the experiment 2.8.3 Determination of protein content is Streptomycin at 20 mg/mL concentration.

The crude proteins were subsequently calculated by multiplying the nitrogen content by a factor of Table 4 shows the Antibacterial Activity of Ethyl 6.25 [24]. The energy value estimation was done Acetate Extract of Medicinal Plants at 20 mg/mL, by summing the multiplied values for crude 40 mg/mL, and 60 mg/mL Concentration. The protein. medicinal plants used for this bioassay are Acacia albida, Anchomanes difformis, Boscia 2.8.4 Determination of crude fat and senegalensis, Bridelia ferruginea, Ficus ingens, carbohydrate content Indigofera arrecta, Moringa olefera and the clinical organism are Salmonella typhi Crude fat and carbohydrate respectively a Water Staphylococcus aureus, Klebsiella pneumoniae, Factors [25,26]. Crude fats were determined by Escherichia coli. The control for the experiment is Soxhlet apparatus using n-hexane as a solvent. Streptomycin at 20 mg/mL concentration.

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Table 2. Antibacterial activity of aqueous extract of medicinal plants at 20 mg/ml, 40 mg/ml, and 60 mg/ml concentrations

Selected clinical Salmonella typhi Staphylococcus aureus Klebsiella pneumoniae Escherichia coli Streptomycin organisms (control) Extracts 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL Acacia albida 10.0 12.0 13.0 10.0 12.0 14.0 10.0 12.0 13.0 4.0 7.0 10.0 2.50 Anchomanes 10.0 11.0 12.0 12.0 15.0 16.0 10.0 11.0 12.0 3.0 5.0 7.0 2.40 difformis Boscia 7.0 8.0 10.0 10.0 12.0 13.0 8.0 10.0 11.0 7.0 8.0 10.0 2.0 senegalensis Bridelia 9.0 12.0 13.0 13.0 14.0 15.0 5.0 7.0 8.0 3.0 6.0 8.0 3.0 ferruginea Ficus 7.0 14.0 15.0 10.0 15.0 19.0 5.0 8.0 10.0 6.0 7.0 8.0 2.7 ingens Indigofera 6.0 10.0 13.0 10.0 11.0 12.0 7.0 9.0 9.5 2.0 6.0 7.0 3.5 arrecta Moringa 6.0 7.0 10.0 7.0 10.0 12.0 4.0 5.0 7.0 5.0 8.0 10.0 4.0 olefera. Unit of zone of inhibition- mm

Table 3. Antibacterial activity of ethanol extract of medicinal plants At 20 mg/mL, 40 mg/mL, and 60 mg/mL concentrations

Selected Clinical Salmonella typhi Staphylococcus aureus Klebsiella pneumoniae Escherichia coli Streptomycin organisms (control) Extracts 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL Acacia albida 2.0 5.0 6.0 1.0 2.0 4.0 3.0 5.0 7.0 2.0 3.0 4.0 3.0 Anchomanes 3.0 5.0 7.0 1.0 4.0 6.0 2.0 5.0 8.0 1.0 3.0 8.0 2.9 difformis Boscia senegalensis 2.0 2..0 5.0 1.0 3.0 5.0 2.0 4.0 5.0 2.0 3.0 6.0 4.5 Bridelia ferruginea 5.0 6.0 6.0 2.0 4.0 6.0 3.0 3.5 5.0 2.0 4.0 7.0 4.0 Ficus Ingens 8.0 10.0 11.0 10. 12.0 13. 5.0 6.0 9.0 8.0 10.0 11.0 5.0 Indigofera arrecta 10.0 11.0 13.0 10.0 11.0 12.0 7.0 8.0 10 2.0 7.0 8.0 4.0 Moringa oleifera. 8.0 10.0 12.0 10.0 11.0 13.0 4.0 5.0 8.0 1.0 5.0 7.0 3.5 Unit of zones of inhibition-mm

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Table 4. Antibacterial activity of ethyl acetate extract of medicinal plants at 20 mg/ml, 40 mg/ml, and 60 mg/ml concentration

Selected Clinical Salmonella typhi Staphylococcus aureus Klebsiella pneumoniae Escherichia coli Streptomycin organisms (control) Extracts 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL Acacia albida 4.0 7.0 9.0 1.0 2.0 3.0 4.0 7.0 9.0 4.0 7.0 9.0 1.0 Anchomanes difformis 1.0 1.0 8.0 1.0 3.0 5.0 3.0 6.0 6.0 1.0 1.0 8.0 5.0 Boscia senegalensis 1.0 1.0 4.0 1.0 4.0 8.0 2.0 3.0 4.0 1.0 3.0 4.0 8.0 Bridelia ferruginea 3.0 3.0 5.0 2.0 3.0 5.0 1.0 2.0 7.0 3.0 4.0 6.0 7.0 Ficus ingens 1.0 1.0 2.0 1.0 5.0 8.0 1.0 4.0 8.0 1.0 4.0 5.0 5.0 Indigofera arrecta 1.0 4.0 8.0 1.0 4.0 7.0 1.0 3.0 6.0 1.0 4.0 8.0 5.0 Moringa oleifera. 1.0 7.0 9.0 1.0 4.0 8.0 1.0 4.0 8.0 1.0 7.0 9.0 8.0 Unit of zones of inhibition-mm

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Table 5 shows the Antibacterial Activity of 4. DISCUSSION Hexane Extract of Medicinal Plants at 20 mg/mL, 40 mg/mL, and 60 mg/mL Concentration. The The pediatrics hospital, little or nothing is known medicinal plants used for this bioassay are about the presence of microorganism in this Acacia albida, Anchomanes difformis, Boscia health care facility, but with this present study, it senegalensis, Bridelia ferruginea, Ficus ingens, is a fact that microbes care also be present and Indigofera arrecta, Moringa olefera and the isolated in this health care facility, putting into clinical organism are Salmonella typhi consideration that children are prone to microbe Staphylococcus aureus, Klebsiella pneumoniae, attack even if, they are still in the hospital. Escherichia coli. The control for the experiment is Adequate care must be taken to reduce the Streptomycin at 20 mg/mL concentration. presence of pathogen in the pediatrics hospital to avoid cross contamination and reemergence of

infectious diseases. Table 6 shows the Qualitative Analysis of Phytochemicals Screening of The Medicinal The Aqueous, Ethanol, Hexane and Ethyl Plants. The medicinal plants are Acacia albida, acetate extracts of the leaves, and roots of Anchomanes difformis, Boscia senegalensis, Acacia albida, Anchomanes difformis, Boscia Bridelia ferruginea, Ficus ingens, Indigofera senegalensis, Bridelia ferruginea. Ficus ingens. arrecta, Moringa olefera and the qualitative Indigofera arrecta and Moringa oleifera were parameter are Alkaloid, Cardiac Glycoside, examined for possible properties of antimicrobial Steroid, Anthraquinone, Phenol, Tannins, and phytochemical activities against the selected Saponin and Flavonoids. clinical organisms. The clinical organisms used are Salmonella typhi, Staphylococcus aureus, Klebsiella pneumoniae and Escherichia coli. Table 7 shows Quantitative Analysis of Minerals Present in Dried Powder of Medicinal Plants It was observed that in Aqueous extract of Extract (mg/100 g).The medicinal plants are Anchomanes difformis, Ficus ingens, Moringa Acacia albida, Anchomanes difformis, Boscia oleifera and Boscia senegalensis shows senegalensis, Bridelia ferruginea, Ficus ingens, appreciable activity of antibacterial potency Indigofera arrecta, Moringa olefera. The minerals against the test organism with the zone of that were are Sodium (Na2+), Potassium (K+), 2+ + 2+ inhibition from 10 mg/mL and 19 mg/mL and Calcium (Ca ), Magnesium (Mg ), Zinc (Zn ), Acacia albida, Bridelia ferruginea and Indigofera Iron (Fe2+), Lead (Pb2+), Copper (Cu-), - + arrecta shows lesser activity of antibacterial Manganese (Mn ) and Phosphorus (P2 ). potency between the ranges of 6 mg/mL to 12 mg/mL. This result conform with the finding of Table 8 shows the Quantitative Analysis Of Anti- [27,17], the action of plant against nutrient present In medicinal plants extract In microorganism. percentage (%). The medicinal plants are Acacia Ethanol extracts shows appreciable high albida, Anchomanes difformis, Boscia antibacterial activity in the plant extract of senegalensis, Bridelia ferruginea, Ficus ingens, Anchomanes difformis, Ficus ingens, Indigofera Indigofera arrecta, Moringa olefera. the anti- arrecta and Moringa oleifera while in Acacia nutrient detected are Tannin, Phenol, Phylate, albida, Boscia senegalensis, Bridelia ferruginea Oxalate, Saponin, Flavonoid, Alkaloids,Tannin shows appreciable low zones of inhibition and Phenol. between the ranges of 6 mL/mg to 12 mg/mL. this conform to the fact that plant are probable Table 9 shows the Quantitative Analysis of source of antimicrobial and therapeutic agent Proximate Composition of Medicinal Plants against some nosocomial organisms especially Extract (%). The medicinal plants are Acacia the organism found in pediatrics hospital and albida, Anchomanes difformis, Boscia other hospital related organism, if this is possible, senegalensis, Bridelia ferruginea, Ficus ingens, Medicinal plants serves as a source good Indigofera arrecta, Moringa olefera The antibiotics, then it can be consumed in are fined detectable parameter are percentage form in our day to day activity, like in our coffee composition of Ash, Moisture content, Crude tea, beverages, soup ingredient, food seasons protein, Fat, Fibre and Carbohydrate. and etc. this is a proof evident by [28].

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Table 5. Antibacterial activity of hexane extract of medicinal plants at 20 mg/ml, 40 mg/ml, and 60 mg/ml concentrations

Selected Clinical Salmonella typhi Staphylococcus aureus Klebsiella pneumoniae Escherichia coli Streptomycin organisms (control) Extracts 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL 40 mg/mL 60 mg/ mL 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL 40 mg/mL 60 mg/mL 20 mg/mL Acacia albida 1.0 3.0 4.0 0.0 1.0 6.0 2.0 5.0 8.0 3.0 5.0 10.0 5.0 Anchomanes difformis 5.0 6.0 7.0 4.0 5.0 10.0 5.0 6.0 7.0 4.0 8.0 10.0 4.0 Boscia senegalensis 4.0 6.0 8.0 6.0 8.0 10.0 5.0 6.0 7.0 4.0 8.0 10.0 5.0 Bridelia ferruginea 2.0 4.0 6.0 1.0 7.0 9.0 5.0 8.0 12.0 10.0 12.0 16.0 6.0 Ficus ingens 2.0 5.0 8.0 3.0 5.0 10.0 2.0 6.0 6.0 7.0 10.0 11.0 3.5 Indigofera arrecta 2.0 6.0 6.0 7.0 5.0 11.0 5.0 6.0 7.0 4.0 8.0 10.0 4.0 Moringa oleifera. 5.0 8.0 12.0 8.0 10.0 16.0 2.0 5.0 8.0 3.0 5.0 10.0 4.2 Unit of zones of inhibition-mm

Table 6. Qualitative analysis of phytochemicals screening of the medicinal plants

Medicinal plants used Alkaloid Cardiac glycoside Steroid Anthraquinone Phenol Tannins Saponin Flavonoids Acacia albida + + + - + + + -

Anchomanes difformis + + + - + + + + Boscia senegalensis + + + + + + + - Bridelia ferruginea - + + + + + - + Ficus ingens + + - + + + + - Indigofera arrecta + + + + + + + - Moringa oleifera. - + - + + + + + Key- Positive (+), Negative (-)

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Ethyl acetate plant extracts shows high zones of The activities of all the medicinal plants extracts, inhibition between 7 mg/mL and 10 mg/mL in the predicts the presence of antimicrobial evidence plant extract of Anchomanes difformis, Indigofera in the medicinal plants under study, it also arrecta while Acacia albida, Boscia senegalensis, brighten the hope for the advents of new Bridelia ferruginea and Ficus ingens shows low antibiotics which can be derived from medicinal zone of inhibition between 1 mg/mL and 5 plants source but not only new antibiotics but mg/mL. Anchomanes difformis and Indigofera evidence for new drugs for the cure of different arrecta are good source of inane booster in the infection associated with pathogen micro- case of HIV patient and some researchers are in organism, under which gram positive and gram the advent of making a good HIV drugs from the negative bacteria has been adequately taken into duo of Anchomanes difformis, and Indigofera consideration [2]. arrecta. This another proof that medicinal plant is a good source of therapeutic agent [29,30]. Ogundare et al. [31] shows that plant species of Anchomanes difformis, Indigofera arrecta Hexane plant extracts shows appreciable and probable V. tenoreana containing saponins, antibacterial activity. Anchomanes difformis, flavonoids, tannins and anthraquinones which Boscia senegalensis, Bridelia ferruginea, were found to have very potent antibacterial Indigofera arrecta and Moringa oleifera shows as well as antifungal activities. These higher zones of inhibition compare with low phytochemical constituents were further zones of inhibition in exhibited by Acacia albida reported to be responsible for many and Ficus ingens. antimicrobial activities of different plant species [32,1].

Table 7. Quantitative analysis of minerals present in medicinal plants extract (mg/100 g)

Medicinal Plants Na2+ K+ Ca2+ Mg+ Zn2+ Fe2+ Pb2+ Cu- Mn- P2+ sample used Acacia albida 22.73 29.11 30.45 26.05 28.04 7.88 ND 0.03 6.00 24.51 Anchomanes difformis 20.56 22,34 21.89 22.10 20.89 5.92 ND 0.04 6.34 25,91 Boscia senegalensis 14.59 14.20 18.77 22.10 21.59 22.10 2.24 1.20 6.23 17.90 Bridelia ferruginea 19.00 23.98 22.12 23.34 16.89 22.12 3.56 2.16 5.79 18.78 Ficus ingens 20.92 23.12 29.34 24.78 17.34 20.34 2.78 3.00 4.92 27.64 Indigofera arrecta 21.33 43.21 16.50 26.37 17.75 4.36 ND ND 14.33 88.43 Moringa oleifera. 20.00 26.14 31.46 22.05 25.03 5.77 ND 0.01 6.20 25.12

Table 8. Quantitative analysis of anti- nutrient present in medicinal plants extract in percentage (%)

PARAMETERS Acacia Anchomanes Boscia Bridelia Ficus Indigofera Moringa albida difformis senegalensis ferruginea ingens arrecta oleifera Tannin 2.20 2.10 2.32 2.37 2.30 2.25 ND Phenol 2.50 3.57 2.56 2.49 3.49 3.45 ND Phylate 16.30 15.27 15.68 15.79 12.33 12.45 2.25 Oxalate 3.66 3.74 6.57 6.51 8.51 8.55 1.50 Saponin 13.87 14.04 9.78 9.70 7.52 7.61 1.67 Flavonoid 8.59 8.52 6.43 6.56 10.38 10.41 ND Alkaloids 3.23 1.25 4.25 4.31 4.36 4.37 Phenol 2.50 3.57 2.56 2.49 3.49 3.45 ND

Table 9. Quantitative analysis of proximate composition of medicinal plants extract (%)

Medicinal plants used % Ash % MC % CP % Fat % Fibre %CHO Acacia albida 10.67 9.31 13.33 6.43 11.49 46.00 Anchomanes difformis 11.69 9.46 12.87 6.46 11.41 47.12 Boscia senegalensis 9.37 3.74 14.68 7.22 4.33 60.55 Bridelia ferruginea 9.39 3.79 14.72 6.82 4.46 60.90 Ficus ingens 8.72 7.32 16.25 5.34 8.53 53.72 Indigofera arrecta 8.78 7.33 16.19 5.40 8.50 53.79 Moringa oleifera 10.53 9.00 14.45 6.50 10.37 42.00 Keys: MC=Moisture Content; CP=Crude Protein; CHO=Carbohydrate

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Flavonoids have been reported to be activities, this important were x-rayed in this synthesized by plants in response to microbial research work. infections and are good antibacterial agents; tannins have been demonstrated to have 5. CONCLUSION antibacterial activities [33,6]. Nigerian populace are fed-up with the foreign Proximate composition of the medicinal plant conventional drugs which can be, and always under study, shows that, carbohydrate, moisture, been adulterated by some disgruntle elements in fat, fibres, ash and crude protein are all present the society, but, there is need for the herbal inclusive in plant analysis. It was observed that, drugs from our natural environment must be carbohydrate has the highest proportion followed developed. This herbal drugs cannot be by crude protein. Scientifically, two proximate adulterated, but can be improved upon, by the quantities are the most importance element in standardizing the production and herbal practice, life. Plants are the medicinal intrinsic worth due to the fact that medicinal plants are all over providing essential nutrients. Proteins and the four geo-political zone of the country, in other carbohydrates are important nutrients to be word the production and the use of the medicinal assessed in medicinal plants [8]. All fractions plant must be standardized and encouraged (cellulose, lignin, hemicellulose, pectin, gums and which can also be a source of revenue to the mucilage) of the fiber play a significant role Nigerian. The medicinal plants have an treating various diseases such as diabetes, antibacterial and phytochemical effectiveness on constipation, diverticulosis, cardio-vascular the tested bacterial isolates resulted within 24 diseases, and fatness, [8,21]. In recent years, hours of incubation of the organism to all the much work has been performed for producing plant extract. The aqueous, ethanol and ethyl and evaluating various sources of proteins and acetate extract display a high level of fibers, [29]. Proteins enclose essential amino antibacterial activity compared to the extract of acids and have the nutritional values for human hexane, but all the plant tested against the health. clinical organism have a certain degree of antibacterial activity, the best activity were found The importance of phytochemicals must be in Anchomanes difformis in aqueous extract, clearly emphases in this present study. Phenols Moringa oleifera in ethanol and ethyl acetate is another phytochemical that were found in extract and Indigofera arrecta in hexane extract the medicinal plant, they are weakly acidic, have little or less activity against the organism, Hydroxyl group attached directly to an knowing fully well, that the hexane extract do not aromatic ring, it serves as Antiseptic, anti- really do well in this research work, but in all, all inflammatory, antimicrobial, anti-tumor and plant were proven active because of their zones also a good disinfectant in its activity which is of inhibition. this result shows that medicinal found in table 6 [31]. plants have a great effect on clinical organism found on pediatric hospital in Ekiti and Ondo Flavonoids, a water soluble, super antioxidant state and free radical scavenger as antioxidant, anticarcinogens, antimicrobial, antitumor. CONSENT Saponins were observe to have bitter taste, foaming property, haemolytic effect on red It is not applicable. blood cells and emulsifying agent. It was also observed that saponin containing plants are good ETHICAL APPROVAL expectorant, cough suppressant, hemolytic activity [34]. It is not applicable.

Tannins were observable in medicinal plant ACKNOWLEDGEMENTS under study.they have unpleasant taste, tans leather, used in the production of leather and The Pediatrics in the Federal Medical Center ink, in treating wounds, varicose ulcers, (Pediatrics) (FMC), Owo, Ondo State, and Ekiti hemorrhoids, frostbite and burns, and it has a State Teaching Hospital (Pediatrics), Ekiti State, soothing relief, regenerates skin, anti- Nigeria. Department of Plant Science and inflammatory and diuretics in its activity [34]. In Biotechnology, Adekunle Ajasin University, other word phytochemical and medicinal plants Akungba Akoko, Ondo State, Nigeria and plays unquantifiable role in our day to day Department of Botany, Ekiti State University Ado

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