Biotechnol Res 2016; Vol 2(2):69-76 Biotechnol Res.2016; Vol 2(2):69-76 eISSN 2395-6763 Copyright © 2016 Epidi et al This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. ORIGINAL RESEARCH Phytochemical, antibacterial and synergistic potency of tissues of Vitex grandifolia 1 1 1 Justice Oyindeinyefa EPIDI , Sylvester Chibueze IZAH *, Elijah Ige OHIMAIN and Timothy T. EPIDI 2 1Department of Biological Sciences, Faculty of Science, Niger Delta University, Wilberforce Island, Bayelsa State, Nigeria 2Department of Crop Science, Faculty of Agriculture, Niger Delta University, Bayelsa State, Nigeria *Corresponding Author email: chivestizah@gmail.com • Received: 10 February 2016 • Revised: 19 March 2016 • Accepted: 23 March 2016 • Published: 04 April 2016 • ABSTRACT This study evaluated the antibacterial and synergistic efficacy of extracts of Vitex grandifolia against some pathogenic microorganisms. The V.grandifolia were obtained within the Niger Delta University campus. Extraction was carried out using water, ethanol, methanol and hexane. Standard agar well diffusion method was employed for the sensitivity test. The experiment was arranged in a Completely Randomized Design fashion. Ethanol extracts were superior in most cases with regard to the zone of inhibition. The highest synergistic zone of inhibition for Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus, Salmonella and Proteus species were 17.00mm (root + stem), 14.00mm (root +leaf +stem), 16.00mm (stem +leaf), 16.33mm (stem +leaf) and 17.00mm (root +stem) respectively. Analysis of variance showed that there were significant differences (P<0.05) in efficacy of their different plant tissues against the bacteria used in the study. The presence of phytochemicals such as tannins, saponins, flavonoids, cardiac glycosides and alkaloids is an indication of the antimicrobial potential of the plant.Thus, V. grandifolia could be used to wade off disease conditions caused by the microorganisms under study. KEY WORDS: Antibacterial, Microorganisms, Phytochemical, Synergistic, Vitex grandifolia Introduction leucoxylon, V. pinnata, V. scabra, V. mollis, V. altissima, V. glabrata, V. megapotamica, V quinata have been assessed The genus Vitex belongs to the Verbenaceae family. About 250 species of Vitex genus are found globally (Ganapaty and Vidyadhar, 2005). Of these, only about 36 have been reported (Owolabi et al., 2010). Of the species of Vitex i.e. V. grandifolia is one of the shrubs found in the tropical rain forest of the Niger Delta but predominantly found in the non- for their pharmacological potentials (Owolabi et al., 2010). Rani and Sharma (2013) reviewed the ethno pharmacology, morphology and microscopy, phytoconstituents, clinical studies, pharmacological reports and toxicological properties of the genus Vitex. The vitex genus typically has essential oils, flavonoids, iridoid, glycosides, diterpenoides and ligans (Rani and Sharma, 2013). wetland region of Nigeria. Hence, V. grandifolia is a facultative upland plant (i.e plants with estimated probability In the traditionally setting some species of Vitex are used for of 66–99% and 1–33% occurring in wetland and non-wetland rheumatic pains, sprains, inflammation, as anti-tubercular, respectively). It can be classified as a Meso-phanerophyte anticancer, diuretic, respiratory infections, in migraine, (mostly tall trees with height in the range of 8 – 30 meters). premenstrual About 16 species of Vitex including V. agnuscastus, V. reported that the leaves of V. negundo are used in the negundo, V. rotundifolia, V. trifolia, V. gardneriana, V. treatment of cold in Tamilnadu. Tijjani et al. (2012) reported problems, anti-fungal, and insecticidal (Ganapaty and Vidyadhar, 2005). Hemalatha et al. (2013) ferrugenia, V. cannabifolia, V. doniana, V. polygama, V. 69 | e I S S N 2 3 9 5 - 6 7 6 3 Biotechnol Res 2016; Vol 2(2):69-76 the activity of V. doniana Stem Bark on Peripheral and Materials and methods Central Nervous System of rat. The leaves and stem-bark Vitex grandifolia authentication have insecticidal potentials against Culexquinque fasciatus (Say) and Anopheles gambiae Giles (Diptera: Culicidae) (Azokou et al., 2013). Also, Epidi and Odili (2009) reported that the leaf powder of V. grandifolia had biocidal effect against Tribolium castaneum in stored groundnut Arachis hypogaea. V. grandifolia Gürke which is commonly known as Black plum, Chocolate and berry tree, (Azokou et al., 2013) is found in the Niger Delta region of Nigeria. The leaves of V. grandifolia are about 10–12 cm in length and 5–7 cm in width (Owolabi et al., 2010). Typical information about this plant is scarce in literature. However, based on these characteristics V. grandifolia bears edible fruits which are used in the production of alcoholic drinks; the bark can also be used in described by Owolabi et al. (2010), Lemmens (2008) and Azokou et al. (2013), the plant was identified. the treatment of stomach ache and diarrhea, bronchial complaints, rickets, sore and fever, while the leaves can be Source and preparation of organisms used for the treatment of diabetes mellitus and as a diuretic The microorganisms including S. aureus, P. aeruginosa, in the treatment of high blood pressure; and as medication Bacillus, Salmonella and Proteus species used in this study against colic, infections of the umbilical cord, toothache, were obtained from the stock culture in the Medical rheumatism and orchitis (Owolabi et al., 2010), hence it has Microbiology and Parasitology Department, College of medicinal properties. According to Ere et al. (2014), Health Sciences, Niger Delta University, Nigeria. The purity medicinal plants have been conventionally used for the of the microbes ascertained as follows: S. aureus used was treatment of several disease conditions in humans for streaked in Mannitol salt Agar which showed yellow thousands of years in many parts of the world. pigmentation. It was further grown on Nutrient Agar, the Medicinal plants which are nature’s gift to humanity (Masih et resultant growth on the Nutrient agar was subjected to al., 2014) form the basis of primary health care for a majority biochemical test. Klingon Iron Agar was used for the of people especially in developing countries (Iniaghe et al., confirmatory test of P. aeruginosa, Salmonella and Proteus 2012; Akinmoladun, 2007; Momoh et al., 2014). Statistics spp. The result was compared with those of known taxa shows that about 80% of global population obtain health using the scheme of Cheesbrough (2004). Also, S. aureus, care services from traditional medicine (Fatima et al., 2011; P. aerugionas, Salmonella, Bacillus, and Proteus species Minochecherhomji and Vyas, 2014; Gahlaut and Chhiller, used in this study were confirmed by conducting some 2013). The patronage to traditional medicine is due to biochemical tests such as gram reaction, motility, catalase, inaccessibility of modern drugs to many people in the rural coagulase, oxidase, citrate, urease and indole on the areas and economic factor (Amole and Ilori, 2010). organisms using the scheme of Benson (2002) and Medicinal plants are essential in the field of pharmacognosy Cheesbrough (2004). probably due to their clinical constituents (Devanaboyina et al., 2013). Antimicrobial screening of the extract Agar diffusion technique was used for the sensitivity test Despite the advancement in the field of microbiology (especially pharmaceutical microbiology), incidence of epidemics due to drug resistant microbes (mainly bacteria and fungi) and the occurrence of emerging and re-emerging microbes still occur (Masih et al., 2014). Several plants have demonstrated antimicrobial properties. Typically, medicinal plants are plants whose tissues including roots, leaves, bark possess healing properties (Minocheherhomji and Vyas, 2014; Kigigha et al., 2015). The above not-withstanding, there is dearth of information on the phytochemical and antimicrobial properties of V. grandifolia in literature hence, the need for this study. using the guide of Opoku and Akoto (2015), Ere et al. (2014) with slight modification using the guide of Kigigha et al. (2015). About 200 μl each of the bacteria was incubated for 24 hours at 37ºC and aseptically inoculated in Mueller Hilton agar plates which were prepared according to the manufacturer’s instruction. The plates were spread using cotton swabs. Four wells of 6.0 mm diameter were made on the agar plates using sterile cock bore, each being for each solvents water, ethanol, methanol and hexane. Approximately 100µl of the concentrated extracts from the different solvents was dispensed into the wells using micropipette. The plates were masked with tape to avoid shifting. Two controls were established using known 70 | e I S S N 2 3 9 5 - 6 7 6 3 Biotechnol Res 2016; Vol 2(2):69-76 sensitive disc. For the sensitivity disc analysis, forceps were statistical analysis. The data were expressed as Mean ± used to pick the disc into solidified agar plates. All the plates standard error. A one-way analysis of variance was carried were incubated at 37ºC for 24 hours under aerobic out at α = 0.05, and Duncan Multiple Range (DMR) Test was conditions. Thereafter, the resultant zone of inhibition was used for the multiple comparison. measured using a meter rule. Results and Discussion Phytochemical screening of the plants Bioactive constituents including Saponins, phlobatannin, Table 1 presents the result of phytochemical screening of V. Cardiac glycosides, alkaloids, flavonoids and tannins were grandifolia. The phytochemical assessment showed that V. determined using the scheme provided by Sofowora (1993), grandifolia contains saponin, alkaloids and flavonoids in all Harborne (1973), Trease and Evans (1993), Okwu (2005) the different plant tissues. Apart from the root of V. cited in Doherty et al. (2010). grandifolia, tannins were found in other tissues of the plant. Statistical analysis and absent in the stem. While, phlobatannin was absent in SPSS software version 16 was used to carry out the the root but present in the stem and leaves. Similarly, cardiac glycoside was present in root and leaves Table 1: Phytochemical constituent of V. grandifolia Tannins Root - Stem + Leaves ++ Saponins Flavonoids ++ + ++ + ++ + Cardiac Glycoside + + Alkaloids ++ ++ + Phlobatannin + + ++= Highly present; + = Moderately present; - = Absence The age and part of the plant play important role in the expectorant, assessment cough suppressant (Osuntokun and The Oluwafoise, 2015). Scientists have variously reported the phytochemicals found in the tissues of V. grandifolia are medical importance of tannins for the treatment of wounds, comparable to those found in other plants as shown for varicose ulcers, hemorrhoids, frostbite and burns (Doherty et of their bioactive constituents. example by the work of Doherty et al. (2010) and Kigigha et al., 2010; Osuntokun and Oluwafoise, 2015; Okwu and al. (2015) on Aframomum melegueta (Alligator pepper). The Okwu, 2004, Kigigha et al., 2015). presence of phytochemicals in plants is an indication that Table 2 presents the zone of inhibition of the different tissue they have functions of biological activity. For example, plants extracts of V. grandifolia. The zone of inhibition of the containing alkaloids have mechanisms by which they resist different bacteria in the ethanolic extracts of the different pests including microorganisms (Agu and Thomas, 2012). plant tissues ranged from 12.00 – 15.33 mm (root), 9.00 – Typically, alkaloids and their synthetic derivatives are 13.00mm (stem), 12.67 – 13.33 mm (leaves), 10.00 – effective for the treatment of analgesic, antispasmodic and 17.00mm (root + stem), 9.33 – 14.00 (root + leaves), 12.33 – bactericidal orientated diseases (Doherty et al., 2010; Opoku 16.33 mm (stem and leaves) and 11 .67 – 13.33 mm (stem+ and Akoto, 2015; Osuntokun and Oluwafoise. 2015). root+ leaves). Typically, there were significant differences Flavonoids which are hydroxylated phenolic compounds help (P<0.05) among the bacteria tested in each of the plant plants to resist disease causing microbes (Opoku and Akoto, tissues and their combinations. The synergistic potency of 2015). Flavonoids have several medicinal properties the different plants tissues showed that the highest zone of including antioxidant, anticarcinogens, antimicrobial and inhibition of 16.33 mm (Salmonella sp), 16.00mm (Bacillus antitumor properties, while the presence of saponin in plant sp), 14.00mm (P. aeruginosa), 14.00mm (Proteus sp) and is an indication that such a plant could be used as 17.00mm (S. aureus) were obtained from stem + leaves, 71 | e I S S N 2 3 9 5 - 6 7 6 3 Biotechnol Res 2016; Vol 2(2):69-76 stem + leaves, root + leaves, root + stem and root + stem respectively. However, there were significant variations (P<0.05) in the highest zone of inhibition that was achieved by each of the bacterium under study. Table 2: Zone of inhibition by different bacteria for the various extracts of V.grandifolia Plant Microorganisms Ethanol Methanol Hexane Water 14.33±0.33fghi 12.00±0.58bcdefgh 0.00±0.00a 0.00±0.00a tissues Root Bacillus sp. S.aureus 14.00±0.00abcd 13.67±0.88fghij 10.67±0.33bcde 9.00±0.58b P.areugenosa 12.33±0.88cdef 13.00±0.58defghij 10.67±0.88bcde 11.33±0.88cdefg 12.00±1.15bcdef 10.67±0.88bcd 13.33±0.88def 15.33±0.88i 15.33±0.88ghij 12.33±0.67cdefg 12.33±0.88bcde 11.00±0.58bcdefg 9.00±0.58a 11.00±0.58bcde 10.00±0.58bc 10.00±0.58bcde 13.00±0.58defg 13.33±0.88efghij 0.00±0.00a 10.00±0.58bcde P.areugenosa 12.67±0.33cdefg 15.00±0.58ij 11.33±0.88bcde 12.33±0.88fg Proteus sp. 12.00±0.58bcdef 11.00±0.58bcde 12.33±0.88bcde 9.33±0.88bc Salmonella sp. 12.00±0.58bcdef 10.67±0.88bcd 10.33±0.88bcd 11.00±0.58bcdef Bacillus sp. 13.33±0.88defgh 10.00±0.58bc 10.00±0.58bc 10.33±0.88bcdefg 13.00±0.58defg 11.33±0.88bcdef 10.00±0.58bc 10.00±0.58bcde P.areugenosa 12.67±0.88cdefg 12.00±0.58bcdefgh 11.33±0.88bcde 11.67±0.88defg Proteus sp. 12.67±0.88cdefg 9.67±0.88b 11.33±0.88bcde 10.33±0.88bcdef Salmonella sp. 12.67±0.88cdefg 11.33±0.88bcdef 11.00±0.58bcde 9.67±0.88bcd 10.00±0.58abc 12.67±0.88defghi 9.33±0.67b 0.00±0.00 a 17.00±0.58j 13.00±0.00defghij 11.00±0.00bcde 11.00±0.00bcdefg P.areugenosa 13.33±0.33defgh 11.00±0.58bcde 13.33±2.85def 0.00±0.00 a Proteus sp. 14.00±0.58efghi 15.00±0.58ij 17.00±0.58g 14.33±0.33 hi 11.33±0.88abcde 12.00±1.15bcdefgh 10.00±0.00bc 10.33±0.33bcdef 9.33±0.33ab 11.67±0.67bcdefg 0.00±0.00a 0.00±0.00a S.aureus 14.00±1.15efghi 14.33±0.33hij 13.67±1.76def 9.33±0.33bc P.areugenosa 14.00±0.58efghi 11.00±1.15bcde 10.33±0.33bcd 10.67±0.67bcdef Proteus sp. 12.00±1.00bcdef 0.00±0.00a 0.00±0.00a 10.67±0.67bcdef Salmonella sp. 13.67±0.88defghi 14.00±1.15ghij 11.67±1.20bcde 11.33±0.88cdefg 16.00±0.58hij 13.67±0.67fghij 12.33±0.88bcde 0.00±0.00a S.aureus 14.00±0.58efghi 13.33±0.33efghij 16.00±0.58fg 13.00±1.15gh P.areugenosa 13.33±2.03defgh 11.00±0.58bcde 0.00±0.00a 12.00±0.00efg 12.33±0.33cdef 15.33±0.88j 0.00±0.00a 0.00±0.00a 16.33±0.88ij 13.00±0.58bcdefghij 13.00±0.58cde 13.00±0.58gh Bacillus sp. 11.67±1.20bcdef 12.67±1.45defghi 11.33±0.88bcde 0.00±0.00a S.aureus 13.33±0.33defgh 12.00±0.58bcdefgh 13.00±1.15cde 9.33±0.33bc 13.00±1.15defg 14.00±0.58ghij 12.67±1.45cde 10.67±0.33bcdef 12.67±0.67cdefg 11.33±0.33bcdef 0.00±0.00a 11.00±0.58bcdefg Proteus sp. Salmonella sp. Stem Bacillus sp. S.aureus Leaves S.aureus Root+ Stem Bacillus sp. S.aureus Salmonella sp. Root+Leaves Bacillus sp. Stem+Leaves Bacillus sp. Proteus sp. Salmonella sp. Root+Stem+Leaves P.areugenosa Proteus sp. 12.33±0.88cdef 12.00±0.58bcdefgh 12.00±1.15bcde 12.00±0.58efg Salmonella sp. Each value is expressed as mean ± standard error (n = 3). Means followed by the same letters in each column is not significantly different at P< 0.05 according to the Duncan Statistics 72 | e I S S N 2 3 9 5 - 6 7 6 3 Biotechnol Res 2016; Vol 2(2):69-76 The zone of inhibition of the different bacteria from the in each of the plant tissues and its combinations. The methanolic extracts of the different plant tissues ranged from synergistic results of the different plants tissues showed that 10.67 – 13.67 mm (root), 10.67 – 13.33mm (stem), 9.67 – the highest zones of inhibition of 13.00 mm (Salmonella sp), 12.00 mm (leaves), 11.00 – 15mm (root + stem), 11.00 – 10.00mm (Bacillus sp), 12.33mm (P. aeruginosa), 15.33 mm 14.33mm apart from Proteus sp that was resistant i.e. (Proteus sp) and 13.00mm (S. aureus) were obtained from 0.00mm (root + leaves), 11.00 – 15.33 mm (stem + leaves) leaves+stem, and 11.33 – 14.00 mm (stem+ root+ leaves). There were respectively. Significant variations (P<0.05) existed in the significant difference (P<0.05) amongst most of the bacteria highest zone of inhibition that was achieved in each of the in each plant tissue and its combinations. The combinations bacterium under study. of the different plants tissues showed zones of inhibition of The significant differences (P<0.05) that existed among the stem, stem, root and stem+leaves 14.00 mm (Salmonella sp), 13.67 mm (Bacillus sp), different tissues or their combination against the selected 15.00mm (P. aeruginosa), 15.33mm (Proteus sp) and 14.33 microbes used in this study could be due to variation in their mm (S. aureus) in respect of root+ leaves, leaves + stem, bioactive constituents as well as the biochemical make-up of stem, leaves + stem and root + leaves respectively. There the different microbes including their metabolism, nutrition were significant variations (P<0.05) in the highest zone of and physiology. Also, the age of the plant and season of the inhibition that was achieved in each of the bacterium under study could also be contributing factors to the variation. study. Generally the ethanol extracts had superior zone of The zone of inhibition of the different bacteria from the inhibition. However, there were fluctuations between the hexane extracts of the different plant tissues ranged from highest zone of inhibition among the organisms as well as 10.67 – 13.33 mm apart from Bacillus sp that was resistant extracts (i.e. solvents). Of all the solvents used, ethanol had (root), 10.33 – 12.33mm apart from S. aureus that was better results though there was fluctuation among the (stem), 10.00 – 11.33 mm (leaves), 9.33 – different solvents used. The trend agrees with the work of 17.00mm (root + stem), 10.33 – 13.67 mm apart Proteus and Masih et al. (2014), Ere et al. (2014), Akintobi et al. (2013). Bacillus sp (root + leaves), 12.33 – 16.00 mm apart from P. The presence of insoluble active compound found in cold aeruginosa and Proteus sp that were resistant (stem and water and or denaturation of the active constituents by the leaves) and 11.33 – 13.00 mm apart from Proteus (stem+ hot water extraction process could be the reason for the root+ leaves). There were significant differences (P<0.05) lower zone of inhibition (Opoku and Akoto, 2015). resistant among most of the bacteria in each of the plant tissues and its combinations. The combinations of the different plant tissues showed that the highest zones of inhibition of 13.00 mm (Salmonella sp), 12.33mm (Bacillus sp), 13.33mm (P. aeruginosa), 17.00 mm (Proteus sp) and 16.00mm (S. aureus) were obtained from leaves+ stem, stem + leaves, stem + root, root+ stem and stem + leaves respectively. There were significant variations (P<0.05) exist in the highest zone of inhibition that was achieved in each of the bacterium under study. The zone of inhibition of the different bacteria from the aqueous water extracts of the different plant tissues ranged The zone of inhibition from this study had some similarity with findings of authors who used different plant species. Kamilla et al. (2009) reported zones of inhibition of methanolic leaf extract (100.00mg/ml) of Clitoria fernatea against some pathogenic microbes including B. cereus, B. subtilis, B. thuringiensis, S. aureus, P. aeruginosa, S. typhi and P. mirabilis as 13.70 mm, 11.30 mm, 10.00 mm, 11.00 mm, 13.30 mm, 21.00 mm and 8.70 mm respectively (leaf), 12.00 mm, 12.00 mm, 14.30 mm, 12.00 mm, 10.70 mm, 18.70 mm and 19.30 mm respectively (stem), 14.00 mm, 12.70 mm, 15.70 mm, 13.00 mm, 11.30 mm, 10.30 mm and from 9.00 – 15.33 mm apart from Bacillus sp (root), 9.33 – 13.70 mm respectively (flower), 12.30 mm, 12.00 mm, 14.00 12.33mm (stem), 9.67 – 11.67 mm (leaves), 10.33 – mm, 12.70 mm, 12.30 mm, 11.30 mm and 15.70 mm 14.33mm apart from P. aeruginosa and Bacillus sp that was respectively (root). Masih et al. (2014) studied the resistant (root + stem), 9.33 – 11.33 mm apart Bacillus sp antimicrobial activities of four plants and reported the zones that was resistant (root + leaves), 12.00 – 13.00 mm apart of inhibition as 29.00 mm and 30.00 mm for S. aureus and P. Proteus and Bacillus sp (stem and leaves) and 9.33 – 12.00 aeruginosa respectively (methanolic extract) and 33.00 mm mm apart from Bacillus sp (stem+ root+ leaves). There were significant differences (P<0.05) amongst most of the bacteria and 26.00 mm for S. aureus and P. aeruginosa respectively (ethanolic extract) for Acacia nilotica, 15.00 mm and 5.00 mm for S. aureusand P. aeruginosa respectively (methanolic 73 | e I S S N 2 3 9 5 - 6 7 6 3 Biotechnol Res 2016; Vol 2(2):69-76 extract) and 10.00 mm and 7.00 mm respectively for S. Akintobi et al. (2013) reported zones of inhibition for different aureus and P. aeruginosa respectively (ethanolic extract) for extracts of Zingiber offinale against S. aureus, P. mirabilis, B. Catharanthus roseus, 14.00 mm and 17.00 mm for S. aureus subtilis, S. typhi and P. aeruginosa as 9.00 mm, 11.00 mm, and P. aeruginosa respectively (methanolic extract) and 0.00 mm, 13.00 mm and 0.00 mm respectively (water 15.00 mm and 14.00 mm for S. aureus and P. aeruginosa extracts) and 13.00 mm, 17.00 mm, 0.00 mm, 10.00 mm and respectively (ethanolic extracts) for Sida cordifolia and 15.00 14.00 mm respectively (ethanolic extracts). However, the mm and 16.00 mm for S. aureus and P. aeruginosa slight variation that exists between this study and previous respectively (methanolic extracts) as 12.00 mm and 15.00 study could be due to age of the plants, type and mm for S. aureus and P. aeruginosa respectively (ethanolic biochemical extract) for Euphorbia birta. Samidurai and Saravanakumar concentration of the plant extracts used as well as the (2009) studied antibacterial activity of methanolic leaf amount of the extract used for the sensitivity test. composition of the different plants, extracts of Pemphisacidula forst and reported the zone of inhibition at different concentrations for S. aureus and P. aeruginosa as 8.00 mm and 6.00 mm respectively (5%), 4.00 mm and 8.00 mm respectively (10%). The zone of inhibitions for the known antibiotics used against S. aureus, P. aeruginosa, Salmonella, Proteus and Bacillus spp. are presented in Figure 1. S. aureus was resistant to Rifampin, Chloramphenicol and Levofloxacin, while the zone Akrayi and Abdulrahman (2013) reported the zone of of inhibition to other antibiotics used including Ciproflox, inhibition of 100% concentration distilled water extract of Norfloxacin, Gentamycin, Amoxil, Streptomycin, thyme, black dry lime and chili as 11.00 mm, 24.00 mm and Erthythromycin and Ampiclox ranged from 21.00 – 27.00mm. 16.00 mm respectively (P. aeruginosa ATCC) and 0.00 mm, P. aeruginosa was resistant to Chloramphenicol and 22.00 mm and 11.00 mm respectively (P. mirabilis). Mann Levofloxacin, while other antibiotics sensitivity ranged from (2012) reported the zones of inhibition of different extracts 16 – 26.00 mm. The sensitivity of Salmonella, Bacillus and (1000ug/disc) of Ocimum gratissimum against S. typhi, P. Proteus spp to all the antibiotics used for the study ranged aeruginosa, S. aureus, P. vulgaris and B. anthracis as 10.20 from 23.00 – 27.00 mm and 22.00 – 27.00 mm, 18.00 – mm, 12.00 mm, 0.00 mm and 0.00 mm respectively 25.00 mm respectively. The variation in the zone of inhibition (ethanolic extract), 12.50 mm, 15.50 mm, 0.00 mm, 0.00 mm could be due to biochemical composition of both the known and 0.00 mm respectively (hexane extract), 0.00 mm, 0.00 antibiotics as well as that of the microorganisms used for the mm, 17.00 mm, 0.00 mm and 0.00 mm (methanol extract). study. Figure 1: Antibiotics sensitivity (mm) results against the bacteria used in this study 74 | e I S S N 2 3 9 5 - 6 7 6 3 Biotechnol Res 2016; Vol 2(2):69-76 Since the extracts of different tissues of V. grandifolia is effective against both gram positive and gram negative bacteria used in this study, it can inferred that they can be used as a broad spectrum antibiotics against bacteria. Some of the known antibiotics reported in this study have been previously reported to be effective against both gram negative and positive bacteria (Pondei et al., 2012; Kigigha Azokou, A., Koné, M. W., Koudou, B. G. and Bi, H. F. T. (2013). Larvicidal Potential of Some Plants from West Africa Against Culexquinquefasciatus(Say) and Anopheles gambiaeGiles (Diptera: Culicidae). Journal of Vector Borne Diseases, 20(2):103–110. Benson, H. J. (2002) Microbiological Applications: Laboratory Manual in General Microbiology. Complete version, 5th edition. McGraw-Hill, New York. Cheesbrough, M. (2004). District Laboratory Practice in Tropical Countries. Low price Edition tissues 2. Cambridge press, England. al., 2009). Devanaboyina, N., Lakshmi, N. R., Satyanarayana, B., Sudeepthi, P., Hemachakradhar, K. and Raju, P. N. (2013). Preliminary Phytochemical Screening, Quantitative Estimation and Evaluation of Antimicrobial Activity of AlstoniaMacrophylla Stem Bark. International Journal of Science Inventions Today, 2 (1): 31-39. Conclusion Doherty, V. F., Olaniran, O. O. and Kanife, U. C. (2010). Antimicrobial activities of AframomumMelegueta (Alligator pepper). International Journal of Biology, 2(2): 126 – 131. et al., 2015; Ere et al., 2014; Samidurai and Saravanakumar, 2009; Amole and Ilori, 2010; Fatima et al., 2011; Kamilla et The use of different tissues of plant for the remedy of several ailments can be dated back to several centuries. The species of plant used by traditional medicine practitioners depend on the availability of such plant in their locality especially the rural dwellers. Microorganisms are ubiquitous and are known to cause several disease conditions. This study investigated the phytochemical, antibacterial and synergistic efficacy of the different tissues of V. gradifolia against disease causing bacteria. 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