International Journal of Research in Pharmacy and Biosciences Volume 1, Issue 1, November 2014, PP 1-4

Phytochemical Content and Antimicrobial Activities of Aegyptiaca ( Gourd) Leaves Extracts

Mankilik M, Mikailu A Directorate of Training and Research Department, Nigeria Institute of Leather and Science Technology (NILEST) Zaria, Nigeria Mhya DH Department of Biochemistry, Ahmadu Bello University, Zaria, Nigeria [email protected]

Abstract: The leaves extracts of Luffa aegyptiaca (Sponge gourd) were screened for phytochemicals, and then investigated for antimicrobial activity by disc diffusion technique. Pulverized leaves of Luffa aegyptiaca was extracted with different solvents by Soxhlet extraction and phytochemicals were determined by color test. Sterile paper disc impregnated with 10 µl of the test extract were aseptically placed onto the surface of the inoculated media while tetracycline (TET) was used as standard. Activities of the extracts were expressed according to zone of inhibition diameter (mm). The phytochemicals analysis of the extracts indicated the presence of some secondary metabolites include cardiac glycosides, flavonoids, saponinis and tannins. The antimicrobial investigations on the extracts against Staphylococcus species, Corynbacterium ulcerans, Bacillus subtilis, Salmonella typhi, E-coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Neisseria gonorrhaeae, and Candida albicans showed good activities with chloroform-extract exhibit the most potent antimicrobial activity. We therefore concluded that Luffa aegyptiaca leaves extract contain bioactive components that exhibit potential antimicrobial activities.

Keywords: Luffa aegyptiaca, Solvent-extracts, Phytochemicals, Antimicrobial activities.

1. INTRODUCTION The utilization of medicines produced from is increasing, thus leading to the interest in the use and importance of medicinal plants as well as intensive research to document scientific evidence for the claimed of their therapeutic efficacy [1, 2, 3]. Plants are known to have substance for self defense against microorganism and the active principle of the medicinal plants are known to be concentrated in organs such as; leaves, stem, root, seed, and fruits among others [4]. Herbs are widely used in Nigeria and other parts of African countries to treat ailments. The folkloric used of medicinal plants could be possibly due to the presence of various chemical agents (phytochemicals) which plays vital protective role against invaded pathogenic microorganism [5]. The ‘Luffa aegyptiaca’ commonly known as sponge gourd belongs to the Curcubitaceace family. It is a vigorous climbing annual with several lobed cucumber-like leaves. The fruits were also cucumber-like shape develops at maturing, with a network of fibers surrounding a large number of flat blackial seeds. It was reported to have been originated from India [6]. It is widely distributed in tropics, subtropics as a cultivated and/or neutralized plant. In Nigeria, Luffa aegyptiaca is grown in almost parts of the country as weed; and it have been reported to posses both medicinal and nutritional potential [6]. Based on this fact, this study wants to investigate and thus report the phytochemical content and antimicrobial activities of the leaves-extract of Luffa aegyptiaca.

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2. MATERIAL AND METHODS 2.1. Materials 2.1.1. Plant Sample Collection and Identification Fresh leaves of Luffa aegyptiaca were collected in Zaria and identified at the Herbarium Unit of the Department of Biological Sciences, Ahmadu Bello University Zaria, Nigeria. 2.1.2. Microbial Bacterial/fungi used for this study were obtained from the stock culture of the Medical Microbiology Unit, Department of Microbiology, Ahmadu Bello University Zaria. The microbial include; Staphylococcus species, Corynbacterium ulcerans, Bacillus subtilis, Salmonella typhi, E-coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Neisseria gonorrhaeae, and Candida albicans. 2.2. Methods 2.2.1. Plant Sample Extraction The leaves of Luffa aegyptiaca were air-dried at 25 oC for 7 days. It was the pulverized using mortar and pestle into fine powdered. The pulverized-leaves were each extracted with aqueous, methanol, ethanol, and chloroform using Soxhlet extractor. The extract was concentrated by allowing evaporating at 30 oC on water bath and then stored in an air-tired sterile container until used. 2.2.2. Phytochemical Analysis Extract of Luffa aegyptiaca leaves was qualitatively analyzed for tannins, flavonoids, saponins, glycosides, steroids, coumarins, anthraquinones and phlobatanins using standard method [7]. 2.2.3. Antimicrobial Testin: To assay for antimicrobial activity of Luffa aegyptiaca leaves extracts, disc diffusion technique was used. Briefly, filter paper (Whatman no. 1) was cut into small sized about 50 pieces and sterilized at 100 oC for 1 hour. About 10ml of the filtered sample of each extract was introduced into conical flasks containing the pieces of paper. The paper were dried at 45-50 oC and the antimicrobial activity of each extract was determined by placing one paper disc at the center of a blood agar plate seeded with test organism. The plate was incubated at 37 oC for 24 hours after which it was examine for growth inhibition. Tetracycline (TET) was used as standard antibiotics; activity of the extract was expressed according to zone of inhibition diameter (mm). 3. RESULTS AND DISCUSSION The physical and phytochemical studies of the leaves extract of Luffa aegyptiaca are presented in Table 1 and Table 2. The phytochemicals analysis indicates the presence of secondary metabolites including tannins, flavonoids, saponins, and cardiac glycosides. However, steroids, phlobatannins, and anthroquinone were not seen in any of the extract analyzed. Luffa aegyptiaca leaves extract contain some natural bioactive compounds that may be of medicinal importance. It have been reported that phytochemical agents plays important role in the treatment of diseases; for instance, cardiac glycosides have found to play significant important role in the treatment of congestive heart failure as well as cardiac arrhythmia [8]. Cardiac glycosides works by inhibiting Na+/K+ pump thereby increasing Sodium ions level in the monocytes this in turn rises the level of Ca2+. Increase in Ca2+ favors the contraction of the heart muscle by improving cardiac output and also reduces distention of the heart [9]. Table 1. Physical properties of Luffa aegyptiaca (Sponge gourd) leave extracts Extracts Indices Methanol (ME) Ethanol (EE) Chloroform (CE) Weight of Extracted (g) 40.00 40.00 40.00 Weight of extract(g) 9.20 9.80 10.00 Percentage yield (%) 23.00 24.50 25.00 Colour Greenish Greenish Greenish Texture powdered powdered powdered

International Journal of Research in Pharmacy and Biosciences 2 Phytochemical Content and Antimicrobial Activities of Luffa Aegyptiaca (Sponge Gourd) Leaves Extracts

Table 2. Phytochemical properties of Luffa aegyptiaca (Sponge gourd) leave extracts Extracts Indices Methanol (ME) Ethanol (EE) Chloroform (CE) Cardiac Glycosides + + + Flavonoids + ++ +++ Saponins ++ + + Glycosides + + + Tannins + ++ ++ NB: + = Mild, ++ = Moderate, +++ = highly present Some bioactive components in plant extracts is known to confer antimicrobial activity against disease- causing microorganism [10] and also offer protection to plants themselves against pathogenic microbial infection [11]. Cowan [12] has earlier reported the antimicrobial activity of some plant phytochemicals. In particular, flavonoids and tannins were reported to be responsible for the antimicrobial properties of some ethnomedicinal plants extracts [9]. Tanins have been known for their astringent property, antimicrobial activity, anti-inflammatory, and anti-diarrhoeal properties [12, 13]. Saponins are known for their medicinal properties as a natural blood cleanser, expectorant and antibiotics [14]. The observed antibacterial activity of the leaves extract of Luffa aegyptiaca is attributed to the presence of bioactive compounds in the extracts. Based on literature reports, we suggested that antimicrobial activity of Luffa aegyptiaca leaves extracts may be due to the presence of some bioactive extracted. In addition, studies of antimicrobial property of Luffa aegyptiaca leaves extract have showed good promising candidate (Table 3). Table 3. Antimicrobial activities of Luffa aegyptiaca (Sponge gourd) leave extract showing zones of inhibition of extract against microorganism (MM) Extracts Antibiotics Indices Methanol (ME) Ethanol (EE) Chloroform (CE) TET Bacillus subtillis 17 19 16 22 Candida albicans 16 18 20 24 Corynbacterium 0 0 0 0 ulcerans E. Coli 20 20 22 25 Klebsiella 20 17 22 24 pneumoniae Neisseria gonorrhaeae 18 22 14 26

Pseudomonas areogen 24 22 27 30 Salmonella typhi 20 22 14 24 Staphyylococus aureus 27 29 30 33 Staphyylococus 17 20 22 30 pyogens Noteworthy to say, the antimicrobial activity of the chloroform extract present the most activity. The activity exhibited by the extracts may be related to the presence and availability of the bioactive agents as suggested above. Yusha’u et al [15] have reported that the presence and availability of phytochemical agents influenced their bioactivity, and this may depends upon method or solvents of extraction employed. 4. CONCLUSION In sum, we can say that Luffa aegyptiaca leaves extract contain good phytochemical agents that have potential antimicrobial properties, and this may be of pharmaceutical importance. ACKNOWLEDGEMENT We wish to acknowledge the supports from the staffs of Medical Microbiology Unit, Department of Microbiology and that of the Herbarium Unit, Department of Biological Sciences, Ahmadu Bello University Zaria.

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REFERENCES [1] M. Hall and N. Nazir (2005). Quantification of the scientific research in United States about popular herbal remedies. Altern: Thes. Health Med., 11:34-37. [2] H.Y. Li, L Cai, and M Cai (2009). Hot topics in Chinese herbal drugs research documented in Pubmed/Medline by authors inside China and outside of China in the past 10 years: Based on Co-word cluster analysis. Journal of Altern. Complement Med., 15: 779-785. [3] I.O. Lawal, N.E. Uzokwe, A.B. Igboanugo, A.F. Adio, E.A. Awosa, J.O. Nwogwugwu, R.A. Adisoga et al (2010). Ethnomedicinal information on collation and identification of some medicinal plants in research Institutes of South-west Nigeria. African Journal of Pharm. Pharmacol., 4(1):001-007. [4] S.L. Kochar (1981). Tropical Crops: A Textbook of Economic Botany. Int College edition, McMillian, London, Uk. [5] S. Renu (2005). Useful metaboliotes from plant tissues culture. Biotechnology, 4(2): 79-93. [6] J.M. Stephen (2003). Gourd Luffa-luffa cylindrical, Luffa aegyptiaca and . Horticul Sci, Univ of Florida, 3: 19-21. [7] A. Sofowora (2008). Medicinal plants and traditional medicine in Africa. 3rd Ed. Spectrum Books limited, Ibadan, Nigeria. Pp:199-204. [8] J. Ngbede, R.A. Yakubu., and D.A. Nyam (2008). Phytochemical screening for active compounds in Cananum schein furthic (Atile) leaves from Jos north, Plateau state, Nigeria. Medical Reseraach Journal of Biological Science, 3(9): 1076-1078. [9] B. Singh and T.K. Bhat (2003). Potential therapeutic application of some anti-nutritional plant secondary metabolites. Journal Agriculture Food Chem. 51:5579-5597. [10] A.C. Farnsworth (2002). The role of ethnopharmacology drug development from plants. England Ciba. John Wiley and Sons, P 2-10. [11] W. De and E. Ifeoma (2002). Antimicrobial components of the bark extract of neem Azadirachta indica A Juss. Technol Dev, 8:23-8. [12] M.M. Cowan (1999). Plant products as anti-microbial agent. Department of Microbiology, Miami University, 12(4):24-29. [13] A.U. Fenglina, L.U. Ruilia, B. Huang, M. Liangh (2004). Free radical scavenging activity of extract prepared from fresh leaves of selected Chinese medicinal plants. Fitoterapia 75: 14-23. [14] E.A. Palomb (2006). Phytochemicals from traditional medicinal plants used in the treatment of diarrhea: Modes of action and effects on intestinal function. Phyto. Rev., 20: 717-24. [15] M. Yusha’u, M.M. Hamza, and N. Abdullahi (2010). Antibacterial activity of Adansonia digitata stem bark extracts on some clinical bacterial isolates. International Journal of Biomedical and Health Sciences, 6(3):129-134.

AUTHORS’ BIOGRAPHY Daniel Hassan Mhya is a holder of BSc & MSc Biochemistry and is currently pursuing PhD Biochemistry at the Department of Biochemistry, Ahmadu Bello Univiersity Zaria, Kaduna state, Nigeria. Mankilik Mary is a senior researcher at the Directorate of Training and Research Department, Nigeria Institute of Leather and Science Technology (NILEST) Zaria, Nigeria. She holds BSc & MSc Biochemistry. Mikailu Abdullahi is a senior researcher at the Directorate of Training and Research Department, Nigeria Institute of Leather and Science Technology (NILEST) Zaria, Nigeria. He holds BSc & MSc Microbiology and is presently pursuing PhD Microbiology at the Department of Microbiology, Ahmadu Bello University Zaria, Kaduna state, Nigeria.

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