Annals of Applied Microbiology & Biotechnology Journal ISSN: 2576-5426

Research Article © Ahaotu EO, et al. 2020

Study on the Anti-microbial and Anti-inflammatory Properties of True Frangipani ( rubra) for the Prevention and Treatment of Diseases in Animal Agriculture Ahaotu E.O1, Nwabueze E2, Azubuike A.P3 and Anyaegbu F3 1Department of Animal Production Technology, Imo State Polytechnic Umuagwo, Nigeria 2Department of Science Laboratory Technology, Imo State Polytechnic Umuagwo, Nigeria 3Department of Agricultural Technology, Imo State Polytechnic Umuagwo, Nigeria

Abstract Plumeria rubra (frangipani) and flowers were extracted using ethyl acetate by maceration process of solvent extraction. The leaves and flowers were screened for phytochemicals, anti-microbial and anti-inflammatory activities. The leaves and flower extracts showed the presence of tannins, alkaloids, balsam, cardiac glycosides, phenols, terpenes and steroids. The extract indicates the absence of flavonoids, saponins and resins. The leaves and flower extracts showed the presence of alkaloids, cardiac glycosides, resins, terpenes and steroids; absence of flavonoids, tannins, saponins and balsam. The zones of inhibition ranges from 10-28mm and the extracts showed a broad spectrum of antimicrobial activity against gram positive and gram-negative bacteria. It was more pronounced on gram negative bacteria especially Proteus mirabilis. Furthermore, the ethyl acetate crude extract was effective against Pseudomonas aeruginosa which is usually resistant to most antimicrobial agents. The extracts were also effective against the fungi Candida albicans. The results of the study can serve as a valuable source of information and provide suitable standards for the prevention and treatment of diseases in animal agriculture.

Keywords: Plumeria rubra; Antimicrobial; Anti-inflammatory; Leaves; Flowers; Minimum inhibitory concentration; Minimum bactericidal concentration; Minimum lethal concentration

INTRODUCTION properties. Odoemelam et al., (2020) indicated that 10kg of Plumeria rubra meal added to the diet of Hy-line brown Plumeria rubra L. a member of family Apocyanaceae is a birds favours hen day production, egg weight, shell weight and common ornamental plant. Distributed throughout the tropics et al., (2020) however reported that the appreciable level of fat in the Plumeria rubra grows up to 15 meters in height. This plant is well known for their feed efficiency of the tested animals. Uduji religiousand cultivated value, nearcosmetic gardens. importance A deciduous and tremendous fleshy stemmed potential tree to be used as medicinal agents to cure infections, digestive reportedflower meal to bebased useful diets as antioxidantas additives andmight hypolipidemic. have accounted Leaves for the egg weight of birds fed these diets. The flowers have been potential, anti-oxidant properties. The plant is also mainly grown are simple, arranged in a whorl, with prominent veins, crowded diseases, anti-inflammatory and antipyretic action, anti-tumor at the end of branches. In traditional medicinal system different parts of the plant have been mentioned to be useful in a variety of The plant material is widely used as a purgative, febrifuge and for its ornamental and fragrant flowers. diseases (Dubey et al., 2014). remedy for diarrhoea and cure for itch. The leaves were reported Flowers are white, reddish pink and bluish with fragrance Plumeria is due to to have analgesic-antipyretic, anti-inflammatory, and antioxidant phenolic compound and is found to be a good source of natural dye Submitted: 05 April, 2020 | Accepted: 20 April, 2020 | Published: 22 April, (Rupali and Alka, 2014). The Pink flowers of 2020 for cloth (Kalam et al., 2014). The fruit is edible, latex is applied to ulcers, herpes and scabies and seeds possess haemostatic *Corresponding author: Ahaotu, E.O, Department of Animal Production properties. Root is bitter, carminative, and thermogenic (Ilyas Technology, Imo State Polytechnic Umuagwo, Nigeria, Email: emmaocy@ yahoo.com et al., et al., Copyright: © 2020 Ahaotu EO, et al. This is an open-access article 2017). 2016).Extract Leavesof leaves are of Plumeria useful in rubra inflammation, rheumatism, distributed under the terms of the Creative Commons Attribution antibacterial, antifungal, bronchitis and antipyretic (Gunja License, which permits unrestricted use, distribution, and reproduction in antibacterial activity against Streptoccocus. Epidermidis and (L.) showed significant any medium, provided the original author and source are credited. Escherischia strains (Singh, 2010). Methanolic extract showed antimicrobial activity against Bacillus anthracis, Pseudomonas Citation: Ahaotu EO, Nwabueze E, Azubuike AP Anyaegbu F (2020) Study on the Anti-microbial and Anti-inflammatory Properties of True Frangipani aeruginosa. The plant is reported to contain amyrinacetate, (Plumeria rubra) for the Prevention and Treatment of Diseases in Ani- mixture of amyrins, ß‐sitosterol, scopotetin, the iriddoids mal Agriculture. Ann Appl Microbiol Biotechnol J 4: 8. doi: https://dx.doi. isoplumericin, plumieride, plumieride coumerate and plumieride org/10.36876/apmbj865282 coumerate glucoside (Egwaikhide et al., 2009).

Ann Appl Microbiol Biotechnol J 4: 8 1/8 1147. 1(1): Res Clin Cardiol J The fruit is reported to be eaten in West Indies. In , Therefore, this study is designed to evaluate the anti-microbial however, it has been used as an abortifacient. The flowers Plumeria are aromatic and bechic and widely used in pectoral syrups. rubra) for the prevention and treatment of diseases in animal The essential oils from the flowers used for perfumery agriculture,and anti-inflammatory test for the properties activities of of true the hexane, frangipani ethyl ( acetate and aromatherapy purposes. Pod has aborti­facient and Plumeria rubra against hepatoprotective effects (Dawada, 2015). Bark is antino­ciceptive four species of Gram negative and ten species of Gram positive and anti-inflammatory. Leaves are found to have antiulcer bacteriaand methanol strains. leaf The and results flower ofextracts the preliminary of phytochemical activity, whereas flowers have profound­ antioxidant effects analysis will provide suggestions as to the probable secondary (Ogunwande et al., 2015). Flower of Plumeria was found to be a metabolites responsible for the activities of the extracts. good source of natural dye for producing various green, ivory and brown shades on silk cloth. MATERIALS AND METHODS The importance of medicinal and aromatic crops in the Collection and authentication national economy and their potential for the rapid growth of Plumeria rubra leaf was collected, from in and around the phyto pharmaceuticals, perfumery and allied industries in Nigeria botany garden of the Forestry Department, Imo State Polytechnic has been emphasized from time to time. Medicinal belong to the oldest known health care products that have been used by plant authenticated specimen is deposited in the Department of mankind all over the world in the form of folklore medicines or ScienceUmuagwo, Laboratory Nigeria Technology and authenticated of the institution. by taxonomist Authentication and the traditional or ethnic medicine (Santhi, 2010). The World Health specimens of the fresh leaves were kept for shade drying. Dried Organization (WHO) estimates that about 4 billion people, 80% specimen was powdered using mechanical grinder and passed of the world population presently use herbal medicine for some through 60 mesh sieve to get the powder of desired coarseness. aspect of primary health care (Reddy et al., 1999). In almost all Powdered material was preserved in an air tight container. and constitute the backbone of the traditional medicine. Extraction of plant material the traditional medicine, the medicinal plants play a major role Medicinal plants are inextricably inter-twined with the rich For preliminary phytochemical analysis, extract was prepared history, culture and culinary tradition of India. India has a rich and glorious ethno medical heritage. The endurance of herbal successive continuous extraction with different solvents as per medicine may be explained often without side effects both theby weighing polarity, 1kgpetroleum of the dried ether, powdered benzene, leafchloroform, was subjected ethanol to andhot on the illness and its symptoms. Various latest technological developments have led to increased accuracy in Estimation, concentratedaqueous. The usingextracts a rotary were filteredevaporator in each at low step temperature using Whatman (40- therapeutically active constituents in crude drugs. Plumeria 45filters0C) andpaper pressure. (Aggarwal The andpresence Paridhavi, or absence 2007). of theThe primary filtrate wasand rubraPurification, commonly Separation known as and Temple Determination tree a small of fugitive principle artistic and secondary Phyto-constituents was detected by usual prescribed tree belongs to the family . It is a small deciduous methods (Dhanapal et al., 2018). Preparation of crude extract havetree with5 cm thickacross, branches white with and yellow copious centre, milky in terminal juice; bark peduncle corky, cymes.fissured. Various Leaves parts 15-30 of cm the long, plant oblanceolate, are useful as thick. medicine. Flowers In powdered with a mechanical grinder and stored in airtight Ayurveda it is used in malarial fevers, antiseptic and stimulant The flowers collected was dried under shade and then (Kumar et al., 2011). The leaves of Plumeria rubra Linn is used with n-hexane and allowed to dry. The product thus obtained wascontainer. then extractedThe dried withpowder methanol material in of a the Soxhlet flowers apparatus. was defatted The bronchitis, cholera, rubifacient, cold and cough (Dhanapal et al., solvent was completely removed under reduced pressure and a in the treatments of ulcer, leprosy, inflammation, rheumatism, 2018). Plumeria rubra Linn plant is traditionally used for the semisolid mass was obtained. treatment of diarrhea, dysentery and typhoid. Microorganisms The emergence of antimicrobial resistant bacteria The two positive bacterial strains Bacillus subtilis, of antimicrobials in any area including disease treatment can Staphylococcus aureus and two negative bacterial strains potentiallypathogens has lead become to widespread a major publicdissemination health concern. of antimicrobial The use Pseudomonas aeruginosa, Escherichia coli including one fungal resistant bacteria. The increasing prevalence of antimicrobial strain Candida albicans are collected for their antimicrobial testing from Department of Science Laboratory Technology, Imo veterinary medicine. Resistant bacteria include both pathogens anddrug-resistant commensal bacteriaorganism, is with a major the later concern serving to as human a potential and Antimicrobial assay reservoir for mobile resistant elements. Since the plant kingdom State Polytechnic Umuagwo, Nigeria. still holds many species of plants containing substances of Disc diffusion method (Garba and Okeniyi, 2010) was used to medicinal values, which are yet to be discovered. Plumeria rubra test the antimicrobial activity of the extracts against four bacterial is one of the plants which have been used in traditional medicine for many years (Oladipupo et al., 2015). paper discs (6 mm diameter) were then impregnated with known strains and one fungal strain (Table 5). Dried and sterilized filter

Ann Appl Microbiol Biotechnol J 4: 8 2/8 Antimicrobial activities ml) and water separately using micropipette and the residual These were done according to the disc diffusion method. For the solventsamount of were the completely test substances evaporated dissolved (Doughari, in methanol 2006). (30 Discs μg/ test, 100 mg of the crude extract of Plumeria rubra was accurately containing the test material with different concentrations each measured by the electronic balance and taken into vial. Then one were placed on nutrient agar medium for bacterial strains and ml of ethanol was added and triturated in uni-directional manner. Sabouraud Dextrose Agar (SDA) for fungal strain uniformly Both gram positive and gram negative bacteria were used. The seeded with the test microorganisms. bacteria used for the anti-microbial activity of Ethanolic crude Negative controls were prepared using the same solvents extract of the Plumeria rubra were Escherichia coli, Salmonella as employed to obtain the extracts. As positive controls, typhi, Salmonella paratyphi, Shigella dysenteriae, Staphylococcus aureus, Streptococcus pyogenes. In this method, measured amount Candida spp. Ciprofloxacin (10 μg/ml) was used for Gram-positive and Gram- negative bacteria and Fluconazole (10 μg/ml) for of the test samples were dissolved in definite volumes of solvent testThe inoculatedmaterials having plates antimicrobialwere incubated activity at 37°C inhibited for 24 h the for growthclinical withto give known solutions amount of known of test concentrationsubstances using (μg/ml). micropipette Then sterile and ofbacterial the microorganisms strains and at and35°C a forclear, 48 distincth for fungal zone strain of inhibition (yeast). wasThe dried.Matricel Standard (BBL, Cookeville, antibiotic USA) discs filter and paperdiscs discson which are impregnated the solvent visualized surrounding the disc. The antimicrobial activity of the used to dissolve the samples is adsorbed and dried were used test agents was determined by measuring the diameter of zone of as positive and negative controls respectively. These discs are inhibition in millimeter. then placed in petri-dishes (120 mm in diameter) containing a suitable agar medium seeded with the test organisms using Physico chemical features sterile transfer loop for anti-microbial evaluation. The plates are The powdered drug was evaluated for its physico-chemical then kept at 40 C for facilitating maximum diffusion. The test parameters like total ash values, acid insoluble ash, water soluble material diffuses○ from the discs to the surrounding medium. The ash and loss on drying, and the results were tabulated (Table 1). plates are then kept in an incubator for 18-24 hours to allow the growth of the microorganisms. The antibacterial activity of the Fluorescence analysis of the extracts test agent is determined by measuring the diameter of the zone The extracts were prepared as per their polarity in hot of inhibition in term of millimeter. It is concluded that the plant successive extraction technique and they were treated with possesses potent antimicrobial activity (Gupta et al., 2008). Anti-inflammatory and anthelmintic activities reagents and the colour changes were observed under Ultra The methanolic extract of Plumeria rubra exhibited VioletExtractive light and values the results were tabulated (Table 3). The extracts were prepared according to the polarity and animal models. The extract (500 mg) exhibited maximum anti- significant anti-inflammatory activity on the tested experimental they were concentrated and their values were calculated with reference to air dried drug and the results were tabulated (Table commonly used as an experimental animal model for acute inflammatory effect. Carrageenan-induced oedema has been 4). method is widely used to evaluate the proliferative components inflammation and is believed to be biphasic. The cotton pellet for detecting the presence of various phyto-constituents present indicated that the methanol extract of P. rubra possess potent All the extracts of the drug was subjected to different tests of the chronic inflammation. The results obtained in this study alkaloids, saponins, proteins, carbohydrate, volatile oil and (Gupta et al., 2006). tannins.in the drug, Preliminary which revealed phytochemical the presence analysis of sterols, indicated flavonoids, a high anti-inflammatory activity in both acute and chronic models of the reasons behind the sedative activity of the plant. These and anthelmintic activity of P. rubra percentage of quercetin and flavonoids and this may be one activityThe saponins was evaluated extract was by used determining for testing theanti-inflammatory reduction in carrageenan induced hind paw edemaleaves. in albino The anti-inflammatory mice. The result parameters, which are being reported for the first time in this P. rubra extract exhibited plantplant, material.are significant towards establishing the Pharmacognostic standards for future identification and authentication of genuine anthelminticof the maximum effect dose of P. ofrubra 200mg/kg isa significant comparable reduction with that in of the the volume effect produced of inflammation. by reference The Table 1: Physico - Chemical Evaluation of the Crude Drug of Leaf of standards piperazine citrateextract on Indian of 25mg/ml adult concentration earthworms Plumeria rubra. (Pheretima posthuma) (Kumar et al., 2012). The chloroform and S/No Physical Evaluation %w/w ethanolic extract of P .rubra leaves shows antiulcer activity in 1. Total Ash albino rats.

2. Acid Insoluble Ash 6.03 The results of the antimicrobial screening have been represented in Table 5. The zone of inhibition of methanol extract Water Soluble Ash 3.942.42 3.4. Loss on Drying 0.5 ranged from 11.88 mm to 26.35 mm. The highest inhibition zone Ann Appl Microbiol Biotechnol J 4: 8 3/8 Table 2. Preliminary Phytochemical Tests for Drug Powder and Various Extracts of Leaf of Plumeria rubra. S. No Test Drug Powder Petroleum Ether Extract Benzene Extract Chloroform Extract Ethanol Extract Aqueous Extract 1. Sterols + + + + + - 2. Terpenoids - - - - + - - - - + +

3.4. FlavonoidsCarbohydrates + + - - - + + 5. Proteins + - - - + + 6. Alkaloids + - - - + + 7. Glycosides ------8. Saponins + - - - + + 9. Tannins + - - - + + 10. Mucilages + - - - + + 11. Volatile Oil + - - - - - + indicates positive reaction, -indicates negative reaction.

Table 3: Fluorescence analysis of leaf of plumeria rubra. ­­­­­­­­­­­­S. No Sample Colour in Day Light Colour in UV Light 1. Petroleum ether extracts Pale green Dark green 2. Benzene Extract Green Light green

3.4. ChloroformEthanol Extract Extract BrownishGreen green GreenDark Green 5. Aqueous Extract Brownish green Yellowish green

Table 4: Extractive values of leaf of plumeria rubra with different solvents. S. No Sample Extractability (%) 1. Petroleum ether extracts 9.5 2. Benzene Extract 7.2

3.4. ChloroformEthanol Extract Extract 5.86.7 5. Aqueous Extract 9.2

Plumeria rubra negative Pseudomonas aeruginosa, Escherichia coli, and fungal against Escherichia coli at the highest concentration followed species Candida albicans was carried out and methanol extract by26.35 Pseudomonas mm was formed aeruginosa, by the Staphylococcusmethanol extract aureus of and Bacillus subtilis. The zone of inhibition of water extract of Plumeria rubra extract of Plumeria rubra Linn. Linn. was less than that of the methanol extract and ranged from showed significant results against pathogens than that of water The results of this investigation should be helpful in the further experiments on antimicrobial activity of Plumeria rubra was formed by water extract of Plumeria rubra against Bacillus subtilis.7.01 mm The to methanol 13.38 mm. extract The highest that showed inhibition antibacterial zone 13.38 activity mm further more mechanistic work is essential to prove it as one of against the pathogens was active in all the given concentration bark. These studies confirms the potential of this plant but

theDISCUSSION specific antimicrobial plant. i.e. 8000, 4000, 2000 and 1000 μg/ml, whereas water extract activityshowed againstactivity Bacillus at the concentration subtilis and Pseudomonas of 8000 and aeruginosa. 4000 mcg/ml but the lower concentrations 2000 and 1000 μg/ml showed no for detecting the presence of various phyto-constituents present Methanol extract as well as water extract of Plumeria rubra All the extracts of the drug was subjected to different tests alkaloids, saponins, proteins, carbohydrate, volatile oil and species, Candida albicans at all the given concentrations (Table in the drug, which revealed the presence of sterols, flavonoids, 5).also In showed vitro antibacterial a significant activity zone of of Plumeria inhibition rubra against bark a against fungal Gram positive Staphylococcus aureus, Bacillus subtilis, Gram thistannins may (Tables be one 2, 3 of and the 4). reasons Preliminary behind phyto-chemical the sedative analysis activity indicated a high percentage of quercetin and flavonoids and

Ann Appl Microbiol Biotechnol J 4: 8 4/8 Table 5. Antimicrobial activity of Plumeria rubra bark Linn. S no. Sample Organism Concentration of Sample Zone Bore Name Diameter Size 1 Methanol Bacillus

Extract subtilis 10mcg\ml 8000mcg\ml (Ciprofloxacin) 25.87mm19.91mm 6mm 4000mcg\ml 18.64mm 6mm 2000mcg\ml 17.24mm 6mm 1000mcg\ml 14.12mm 6mm

Staphylococcus aureus 10mcg\ml (Ciprofloxacin) 28.33mm 6mm 4000mcg\ml 8000mcg\ml 19.58mm 23.11mm6mm 6mm 2000mcg\ml 16.56mm 6mm 1000mcg\ml 12.71mm 6mm

Pseudomonas

Aeruginosa 10mcg\ml 8000mcg\ml (Ciprofloxacin) 28.75mm 24.14mm 6mm 4000mcg\ml 22.08mm 6mm

2000mcg\ml1000mcg\ml 18.36mm15.47mm 6mm

Escherichia

Coli 10mcg\ml (Ciprofloxacin) 30.00mm 6mm 8000mcg\ml 26.35mm 6mm 2000mcg\ml4000mcg\ml 23.17mm16.68mm 6mm 1000mcg\ml 11.88mm 6mm

Candida albicans 10mcg\ml (Fluconazole) 17.36mm 6mm 4000mcg\ml 8000mcg\ml 10.41mm 11.23mm 6mm 6mm 2000mcg\ml 9.15mm 6mm 1000mcg\ml 8.58mm 6mm

2 Water Bacillus extract subtilis 10mcg\ml (Ciprofloxacin) 23.19mm 6mm 4000mcg\ml 8000mcg\ml 9.21mm 13.38mm 6mm 6mm 2000mcg\ml Nil 6mm 1000mcg\ml Nil 6mm

Staphylococcus aureus 10mcg\ml 8000mcg\ml(Ciprofloxacin) 25.91mm 9.22mm 6mm 6mm 4000mcg\ml 8.61mm 6mm 2000mcg\ml 8.01mm 6mm

1000mcg\ml 7.38mm 6mm Pseudomonas

10mcg\ml (Ciprofloxacin) 24.5mm 6mm

Ann Appl Microbiol Biotechnol J 4: 8 5/8 aeruginosa 8000mcg\ml 7.88mm 6mm 4000mcg\ml 7.01mm 6mm 2000mcg\ml Nil 6mm 1000mcg\ml Nil 6mm

Escherichia coli

8000mcg\ml 10mcg\ml (Ciprofloxacin) 8.58mm 31.75mm 6mm 6mm 4000mcg\ml 7.51mm 6mm 2000mcg\ml 7.04mm 6mm 1000mcg\ml Nil 6mm

Candida albicans

8000mcg\ml 10mcg\ml (Fluconazole) 8.22mm 15.36mm 6mm 6mm 4000mcg\ml 8.07mm 6mm 2000mcg\ml 7.62mm 6mm 1000mcg\ml 7.11mm 6mm Kumar et al., (2012). of the plant. These parameters, which are being reported for authenticationthe first time in of thisgenuine plant, plant are material.significant Though towards Plumeria establishing rubra isthe a temple Pharmacognostic tree, it is a highly standards reputed for drug future used identificationfor the prevention and and treatment of diseases in animal agriculture. The literature survey revealed that the various species of Plumeria is an important source of many pharmacologically and medicinally important chemicals such as plumieride, Figure 1a Dorsal View of the leaf. isoplumeride, fluvoplumericin, irriod glycoside and other various minor secondary metabolites. Study of pharmacological activities with different extracts obtained from different parts of the plant a(Figure number 1, of 2 anddiseases. 3) with The difference plant has inbeen vitro widely and instudied vivo model,for its pharmacologicalwhich show that activitiesthe compounds and regarded have beneficial as universal effects panacea against in plant having a wide spectrum of medicinal activities. Plumeria rubraethno - veterinary medicines and find its position as a versatile Figure 1b Ventral view of the leaf. resembling a broad spectrum antibiotic against the common uro- gastro pathogenicappears toEscherichia have significant coli, one of antimicrobial the common capacitybacteria with pathogenic strains and are relatively resistant towards synthetic drugs (Syakira and Brenda, 2010). As the global scenario is now changing towards the use of non-toxic plant products, development of modern drugs from Plumeria species should be emphasized. plumeric acid from the latex, leaves and fulvoplumerin from the bark of P. rubra possess local anesthetic,Ursolic cardiotonic acid from theand leaves, bacteriostatic activities respectively. P.rubra containing fulvoplumerin acts as inhibitors of human revereasetrancriptase. Methanolic extract of P.rubra showed hepatoprotective action immunodeficiencyagainst paracetamol virus induced type hepatic 1 (HIV) damage. Ethanolic extract Figure 1c Flowers. of Plumeria rubra

. (Apocynaceae) leaves and flowers were tested Ann Appl Microbiol Biotechnol J 4: 8 6/8 for antimicrobial activity against Gram-positive bacteria (Bacillus The antimicrobial activity was more pronounced in the gram- subtilis, Enterococcus faecalis and Staphylococcus aureus), Gram- negative Staphylococcus aureus, a gram-negative bacterium. The negative bacteria (Escherichia coli, Klebsiella Pneumonias, plant extract was also effective against the fungi Candida albicans. Pseudomonas aeruginosa, Salmonella typhimurium) and fungi World Health Organization (WHO) has emphasized the (Aspergillus niger and candida albicans) by disc diffusion need to ensure quality control of the raw materials used for method. The ethanol extract showed strong in vitro antimicrobial ethno-veterinary medicines by using modern techniques, by activity against E. faecalis, B. subtilis, S. aureus, P. aeruginosa, S. applying suitable parameters and standards. In the present typhimurium, A. niger and C. albicans respectively (Rasool et al., study various standardization parameters such as macroscopy, 2008). microscopy (histochemical and powder), physicochemical Plumeria rubra extracts were evaluated for antimicrobial standards, preliminary phytochemical investigation, which are activity using cup plate method and minimum inhibitory concentration against Escheria Coli, Bacillus subtilis, authentication and preparation of a suitable monograph for the Staphylococcus aureus and Aspergillus niger. It was observed that being reported for the firstPlumeria time inrubra this forplant the could future. be helpful in The prevention of oxidative damage to tissue could therefore strains. When compared with Ciprofloxacin as a standard, proper identification of aqueousa methanol extract extract was exhibited active againstsignificant fungal activity strains against as compared bacterial with standard Fluconazole (Surendra et al., 2012). be one of the mechanisms responsible for the anti-inflammatory Methanolic extract of P. rubra effect shown by this plant. Confirmation of the anti-inflammatory antitumor activity against Dalton lymphoma ascites in mice P.rubraactivity in animal model further justifies the traditional use of result shows that methanolic extractleaves of P. alba possesses significant this plant for inflammatory disorders. The ethno medical use of prolong the life span, reduce tumour volume and improve the antioxidantas a usefulpotential. remedy in inflammatory and arthritic disorders hematological parameters of the host (mice) (Radhacan significantlyet al., 2008). could possibly be because of its excellent anti-inflammatory and ACKNOWLEDGEMENT CONCLUSION The authors are thankful to the Director of Tetfund Institution It is reasonable from the result obtained to suggest that the – Based Intervention for providing funds and facilities to carry plant extracts possess broad spectrum antimicrobial activity. out the present research. REFERENCES 1. Aggarwal SS, Paridhavi M. Extraction, isolation and analysis of phyto-

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