Tarun Agarwal et al.

/ JPBMS, 2012, 15 (12)

Available online at www.jpbms.info

Research article JPBMS

ISSN NO- 2230 7885 CODEN JPBSCT

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL SCIENCES

Comparative Analysis of Antibacterial Activity of Jatropha curcas Fruit Parts

Dr. Singh Rachana, Agarwal Tarun*, Rastogi Rinki, Arora Neha, Rastogi Meghna.
Amity Institute of Biotechnology, Amity University, Lucknow, India.

Abstract:

The present study presents the effectiveness of Jatropha curcas Fruit against some selected microorganisms which are known to cause diseases in human beings and the comparative study of extent of antimicrobial properties of various fruit parts of Jatropha curcas namely, Fruit Pericarp, Seed Coat and Seed Kernel. Cold Aqueous, Methanolic, Ethanolic, and Ethyl Acetate extracts of dried fruit parts were prepared at a final concentration of 500 mg/ml and were tested against pathogenic microorganisms such as Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli using agar well diffusion method. The Methanolic extracts of the Seed Kernel showed the maximum zone of inhibition of 25.5mm.

Keywords: Antibacterial properties, Jatropha curcas fruit, Fruit Pericarp, Seed Coat and Kernel, Agar Well Diffusion. Introduction:
Widespread use of drugs is leading to the development of resistance against them in the pathogen [1] and also the side effects associated with them is urging people not to use them. Therefore there is a constant and urgent need to develop new antimicrobial drugs for the treatment of infectious disease from medicinal plant [2]. Anti-microbial agents are substances that kill microorganisms or inhibit their growth. They are widely employed to cure bacterial diseases. Antimicrobial agents that reversibly inhibit growth of bacteria are called bacteriostatic whereas those with irreversible lethal action on bacteria are known as bactericidal [3]. Ideally, antimicrobial agents disrupt microbial processes or structures that differ from those of the host. They may damage pathogens by hampering cell wall synthesis, inhibiting microbial protein and nucleic acid synthesis, disrupting microbial membrane structure and function, or blocking metabolic pathways through inhibition of key enzymes [4]. The J. curcas Linnaeus plant originated from Mexico and was spread to Asia and Africa by Portuguese traders as a hedge plant. J. curcas L. belongs to the family Euphorbiaceae. In many subtropical and semi-arid regions, traditionally, J. curcas is used for its medicinal properties and its seeds contained semi dry oil which has been found to be useful for medicinal purposes. It has played a major role in the treatment of various diseases including bacterial and fungal infections. The seeds and leaves extracts of J. curcas, have shown molluscidal and insecticidal properties [5]. They showed that methanol extracts of J. curcas had the highest activity against vector snails of the human parasites Schistosoma spp. [6]. The extracts of many Jatropha species including J. curcas displayed potent cytotoxic, anti-tumor and anti1

microbial activities in different assays. The latex of J. curcas also showed antibacterial activity against Staphylococcus aureus, however the antimicrobial activity of the other parts have not been fully investigated [7]. The objectives of this study was to investigate the effectiveness of Jatropha curcas Fruit against some selected microorganisms which are known to cause diseases in human beings and the comparative characterization of extent of antimicrobial properties present in the fruit parts of Jatropha curcas namely, Fruit Pericarp, Seed Coat and Seed Kernel.
Figure 1: Jatropha curcas Fruit Pericarp (Pale Yellow), Seed Coat (Brown) and Seed Kernel (White)

Materials and methods

Sample Collection The fresh fruits of Jatropha Curcas were collected from Kanpur after proper identification and brought to the laboratory. The Jatropha Curcas fruits were washed with tap water followed be distilled water. The fruits were then dried as a whole. Each part of the dried fruit, namely, fruit pericarp, seed coat and the seed kernel were separated and were grinded into fine powder by the help of motar pestle.

Journal of Pharmaceutical and Biomedical Sciences (JPBMS), Vol. 15, Issue 15

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Bacterial strains and culture preparations Three pathogenic strains namely Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli, available at Amity University, Lucknow were subcultured and used throughout the study. Preparation of Plant Extract Antimicrobial metabolites from the dried fruit parts of Jatropha Curcas were extracted using various solvents such as Cold Aqueous, Methanol, Ethanol and Ethyl Acetate. 2 gm of powdered dried sample was soaked in 20 ml of the respective solvents (1:10) and kept in dark for 45 days so as to let the secondary metabolites to diffuse out into the used solvents. It was then filtered in weighed petri plate and dried in hot air oven at 50oC, so as to evaporate the solvents. The dried metabolite extract was dissolved in double volume of DMSO (Dimethyl Sulfoxide) thus giving the final concentration of extract to 500 mg/ml. Antibacterial Susceptibility Assay Antibacterial susceptibility assay was carried out by well diffusion method [8] wherein sterile Nutrient agar plates were prepared and spreaded with 50ml of the available bacterial cultures against which antibacterial activity was tested. There after 3 wells of 8 mm diameter were dug with the help of sterile borer. Two plates were prepared

for each microbial strain and for each part of the fruit of Jatropha Curcas used in the studies. In the plate 1; the 1st, 2nd and 3rd well was filled with 60l of standard antibiotic Tetracycline, Methanolic and Ethanolic extract respectively. In the plate 2; the 1st, 2nd and 3rd well was filled with 60l of standard antibiotic Tetracycline, Cold Aqueous and Ethyl acetate extract respectively. Plates were incubated at 370C for 24 hours. The antibacterial activity of each extract was expressed in terms of mean of diameter of Zone of Inhibition (in mm) produced by each extract at the end of incubation period.

Results:

Antibiogram analysis: In order to check the antimicrobial activity of extracted plant samples, agar well diffusion method was used. Table 1-3 below, shows the results of zone of inhibitions (ZOI) observed for the antimicrobial properties in the extracts of all the three parts of the fruit namely Fruit Pericarp, Seed Coat and Seed Kernel against the standard antibiotic tetracycline.

Table 1: Antibacterial Susceptibility Assay of Jatropha curcas Fruit Pericarp Extract EXTRACTS ZONE OF INHIBITION (ZOI) AGAINST (in mm) S.NO. Escherichia coli By Extract By Tetracycline Pseudomonas aeruginosa By Extract By Tetracycline Staphylococcus aureus By Extract By Tetracycline

1 2 3 4

Ethanol Methanol Ethyl Acetate Cold Aqueous

20.0

17.0 17.0 18.5 18.5

12.5 12.5 17.5

13.0 13.0 14.0 14.0

19.5 17.5

16.5 16.5 15.0 15.0

Note: Well diameter = 8mm.

Table 2: Antibacterial Susceptibility Assay of Jatropha curcas Seed Coat Extract S.NO. EXTRACTS ZONE OF INHIBITION (ZOI) AGAINST (in mm) Escherichia coli By Extract By Tetracycline Pseudomonas aeruginosa By Extract By Tetracycline Staphylococcus aureus By Extract By Tetracycline

1 2 3 4

Ethanol Methanol Ethyl Acetate Cold Aqueous

16.5 17.0

16.0 16.0 17.0 17.0

21.0 13.5

12.0 12.0 13.5 13.5

18.0 20.0 14.0

17.0 17.0 15.0 15.0

Note: Well diameter = 8mm.

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Table 3: Antibacterial Susceptibility Assay of Jatropha curcas Seed Kernel Extract. S.NO. EXTRACTS ZONE OF INHIBITION (ZOI) AGAINST (in mm) Escherichia coli By Extract By Tetracycline Pseudomonas aeruginosa By Extract By Tetracycline Staphylococcus aureus By Extract By Tetracycline

1 2 3 4

Ethanol Methanol Ethyl Acetate Cold Aqueous

13.0 14.0 13.5 13.0

15.0 15.0 15.5 15.5

12.0 25.0 12.5 11.5

13.0 13.0 13.5 13.5

25.5 12.5

16.5 16.5 14.0 14.0

Note: Well diameter = 8mm.

Discussion:

activity; both antibacterial as well as antifungal properties

[7].

Researchers have extensively studied the biological properties of Jatropha curcas and their results showed that this plant is ethno-medically valuable. The Methanolic extracts of the leaves, roots and the stem have already been shown as to possess the antimicrobial
Figure 2: E. coli: Comparative study of Jatropha curcas extract various solvents.

The figure 2-4 shows the graphical comparisions made on the extent of antimicrobial properties possessed by the three parts of Jatropha curcas fruit against selected microorganisms.
Figure 3: S. aureus: Comparative study of Jatropha curcas extract of extract of various solvents

Figure 4: P. aeruginosa: Comparative study of Jatropha curcas extract of various solvents.

Agar well diffusion method was used here in order to determine the anti microbial properties of the plant extracts against the pathogens performed earlier by [9]. The Methanolic extract of Jatropha curcas Seed Kernel showed the maximum Zone of Inhibition (25.5 mm)
3

against Pseudomonas aeruginosa, followed by Zone of Inhibition (25 mm) of the same against Staphylococcus aureus. The Ethyl Acetate extracts of the Seed Coat showed the zone of inhibition of the 21 mm against Pseudomonas aeruginosa.

Journal of Pharmaceutical and Biomedical Sciences (JPBMS), Vol. 15, Issue 15

Tarun Agarwal et al. / JPBMS, 2012, 15 (12)

The Cold Aqueous extract of all the parts of the Jatropha curcas fruit showed Zone of Inhibition against all the bacterial strains. The Ethanolic, Methanolic, Ethyl Acetate extracts of Jatropha curcas Fruit showed much varied results against all the bacterial strains used in the studies. The Ethyl Acetate extracts of Jatropha curcas possess good antimicrobial properties against Staphylococcus aureus and Pseudomonas aeruginosa. The Ethanolic, Methanolic, Ethyl Acetate and Cold aqueous extracts of Jatropha curcas of the Seed Kernel showed a clear Zone of Inhibition against Pseudomonas aeruginosa and Escherichia coli. But only the Methanolic and Cold Aqueous extracts of the same showed ZOI against Staphylococcus aureus. The antibacterial activity of Fruit of Jatropha curcas may be indicative of presence of metabolic toxins or broad spectrum antimicrobial compounds that act against gram +ve as well as gram ve bacteria especially in solvents Ethanol, Ethyl Acetate, Cold Aqueous and Methanol. Based on the above research work it can be concluded that the fruit of Jatropha curcas can be a very good source for herbal drugs especially in Cold Aqueous, Ethyl Acetate and Methanol as a solvents. This can be explored further for the extraction of antimicrobial agents by more sophisticated procedures for extraction in order to increase the yield.

The future prospects of present research work includes isolation and purification of the therapeutic antimicrobial compounds from the active extract and there further pharmacological evaluation by several method such as NMR, MS, GC-MS, TLC, HPLC.

Acknowledgement:

We are thankful to our HOI, Dr. Rajesh K. Tiwari and the Management of Amity Institute of Biotechnology, Amity University, Lucknow for giving the permission and providing us the facilities to perform the research work. We would also like to the Lab Assistants and staff without whose kind cooperation the working could not have been possible.

References:
1.Davis J, Science, 2009, 264, 375-82. 2.Cordell GA, Phytochemistry, 2000, 55, 463-480. 3. Rajesh B, Rattan LI, Essentials of Medical Microbiology, 4th Edition, Jaypee Brothers Medical Publishers (P) Ltd., New Delhi 110002, India, 2008, 500 pp 4. Arekemase MO, Kayode RMO, Ajiboye AE, International Journal of Biology, 2011, 3(3), 52-59. 5. Rug M, Ruppel A, Trop. Med. Int. Health, 2000, 5, 423430. 6. Daouk RK, Dagher SM, Sattout EJ, J. Food Prot., 1995, 58, 1147-1149. 7. Thomas OO, Fitoterapia, 1989, 60, 147-161. 8. Perez C, Anesini C, J. Ethnopharmacology, 1993, 44, 4146. 9.Khan JA, Kumar N, J. Pharma and Biomed. Sciences, 2011, Vol. 11, 1-3.

Conclusion:

Conflict of Interest: - None Source of funding: - Not declared Corresponding Author:Tarun Agarwal., Amity Institute of Biotechnology, Amity University, Lucknow, India. Address: 566/18 KHA, JAI PRAKASH NAGAR, ALAMBAGH, LUCKNOW, U.P. 226005 Mobile: +919792729363.

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