Indian Streams Research Journal

Volume 2, Issue. 7, Aug 2012

Available online at www.isrj.net

ISSN:-2230-7850 ORIGINAL ARTICLE

In Vitro Antibacterial Activity In The Extracts Of Solanum Nigrum

Parameswari.K1, Sudheer Aluru2 and Kishori. B3

1

.Department of Biotechnology Padmavati Mahila Visvavidyalam, Tirupati - 517 502. A.P. India. 2 .Department of Anthropology, Sri Venkateswara University ,Tirupati - 517 502. A.P. India. 3 Asst. Professor, Dept.of Biotechnology,Sri Padmavati Mahila University Tirupati 517 502. A. P. India. kktinku@rediffmail.com Abstract: The present investigation was carried out to study the anti-bacterial activity of ethanolic and methanolic extracts of stem, berries and whole plant of Solanum nigrum. The antibacterial activity was detected against Bacillus subtilis, Escherichia coli, Klebsiella pneumonia and Pseudomonas aeruginosa. Solanum nigrum plant extracts of, 10g, 50g and 100g were taken as per CLSI standards. The zones of inhibitions obtained were recorded and analyzed against standard control streptomycin. The methanolic extracts were showed highest antibacterial activity than ethanolic extracts. In both extracts, whole plant extract showed potential anti-bacterial activity than stem and berries. The results indicate that the whole plant extracts of Solanum nigrum may recommend to use in preparation of herbal drugs.

KEY WORDS: Antibacterial activity, Solanum .nigrum, stem, berries, whole plant extracts, agar well diffusion. INTRODUCTION Recently, in world 80% of populations are using medicinal plants to cure various diseases caused by micribial pathogens. Increasing the failure of synthetic drugs, side effects and development of antibiotic resistance by pathogenic microorganisms leads to development of the identification and screening of several medicinal plants for their potential antimicrobial activity (Iwu et al., 1999). Thousands of species are known to have medicinal value and several studies reported the antimicrobial activity of plants (Werner et al.,1999; Samy and Ignacimuthu, 2000). In this regard value of traditional medicine has recognized by World Health Organization and involved in creating strategies, guidelines and standards for plant medicines (WHO, 2002). In pharmaceutical industries, due to varied medicinal properties in all plant parts include root, stem, flower, fruit, whole plant and modified plant is used for extract as drugs. So, an extensive study is required to detect the medical properties of the plant. Several medicinal plants have been tried against pathogenic microorganisms (Haraguchi et al., 1999; Sashikumar et al., 2003). In ancient times parts of several medicinal plants are used to cure specific diseases (Hashim et al 2010). But the different parts of several medicinal parts to cure specific ailments have been in vogue from ancient times. The potential of higher plants as source for new drugs is still largely unexplored. Among the estimated 250,000-500,000 plant species, only a small percentage has been investigated phytochemically and the fraction submitted to biological or pharmacological screening is even smaller (Mahesh and Satish, 2008). Several medicinal plants have been tried against pathogenic microorganisms, hence to detect the plant medicinal properties an extensive study is required (Haraguchi et al., 1999; Sashikumar et al., 2003). Solanum species are the most
Please cite this Article as : Parameswari.K1, Sudheer Aluru2 and Kishori. B3 , In Vitro Antibacterial Activity In The Extracts Of Solanum Nigrum : Indian Streams Research Journal (Aug. ; 2012)

In Vitro Antibacterial Activity In The Extracts Of Solanum Nigrum

potent plants against pathogenic microorganisms. Solanum incanum (L) is one of the important traditional medicinal plant belongs to Solanacae family. Antibacterial activity of Solanum incanum was studied (John Britto, 2001; John Britto and Senthilkumar 2001; Pavitra et al., 2012) and presence of analysis of phytochemicals were also studied (Pavitra et al., 2012). Other solanum species, Solanum torvum (leaf, stem and roots) showed antibacterial and antifungal activity (Bari et al., 2010) and antibacterial activity of Solanum surattense whole plant extract (Patil Suhas et al., 2009) and leaf extract (Sheeba, 2010) were studied. Analysis, presence of phytochemicals and potent antibacterial activity of leaf, root and seed extracts were studied in Solanum nigrum (Sridhar et al., 2011). In view of this the present study was aimed to evaluate antibacterial activity of other plant parts like stem, berries and whole plant extracts of Solanum nigrum against to some bacterial pathogens. MATERIALS AND METHODS COLLECTION OF PLANT MATERIALS Fresh Solanum nigrum whole plants and plant parts like stem, berries and whole plant were collected randomly from nurseries in Bangalore. Botanical identification of plant material was done by Department of Botany, Indian Academy group of Institutions. The plant materials were washed thoroughly for 3-4 times with running tap water to remove dust from the surface and twice with sterile distilled water, shade dried at room temperature on a sterile blotting paper, after complete drying the plant materials were powdered using the blender and stored in separate air tight bottles. Then used for preparation of methanolic and ethanolic extracts. PREPARATION OF METHANOL EXTRACT Air-dried 10 g of each powdered plant material was mixed in a conical flask with 100ml of deionised distilled water and methanol, then shaken at 120 rpm for 30 minutes and kept for 24 h. After 24 h, each of the extracts was filtered rapidly through four layers of gauge and then by a more delicate filtration through Whatman no.1 filter paper. The resulting filtrates were then concentrated in a rotary evaporator for 24 hours to collect the Methanol extract. Similar method was used to prepare ethanol extract. TEST MICROORGANISM The four bacterial strains, Bacillus subtilus, Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa were collected from Department of Microbiology, Indian Academy group of Institutions, Bangalore as test bacteria for antibacterial activity assay. ANTIBACTERIAL ACTIVITY DETERMINATION A direct suspension of the test organisms was prepared in broth and the optical density (OD) of the solution was adjusted to 0.5 at 456 nm which corresponded to 1x 107 CFU. The antibacterial activity assay was performed by sensitivity tests by agar well diffusion method. Mueller Hinton Agar was used as media as per CLSI guidelines. The test bacterial strains 100l were inoculated in to Muller Hinton agar medium. After that 6 mm diameter were punched in agar plate. Different concentrations (10g,, 50g, and 100g) of stem, berries and whole plant extracts were poured in the wells. Standard antibiotic streptomycin (10g/ml) and blank (solvent and water) were used as positive and negative control. The plates were then incubated at 37oC for 24 hr to allow maximum growth of the microorganisms the antibacterial activity of the test samples was determined by measuring the diameter of zone of inhibition expressed in millimeter. The assay was repeated thrice and the mean of the three experiments was recorded. RESULTS AND DISSCUSION The results of antibacterial activities of the Soanum nigrum were shown in Table 1. The extracts showed varying degree of inhibitory effect. The inhibition zone of extracts was directly proportional to increasing concentration of field grown different extract of S. nigrum. Whole plant extracts showed highest antibacterial activity in compared with stem and berries in both methanolic and ethanolic extracts (Table 1). The small zone of inhibition was observed in 10g concentration of whole plant ethanolic extract against to Bacillus subtilis. No zone of inhibition was observed against to E coli, Klebsiella pneumonia and Pseudomonas aeruginosa. But zone of inhibition was observed in 10g concentration of methanolic
Indian Streams Research Journal Volume 2 Issue 7 Aug 2012

In Vitro Antibacterial Activity In The Extracts Of Solanum Nigrum

extract of stem and berries, against to all bacteria. Stem and berries showed minimum zone of inhibition (3.0mm 9.0mm) with 10g concentration of ethanolic extract against to tested microorganisms. Methanolic extracts of stem, berries and whole plant were showed highest antibacterial activity than ethanolic extracts. Maximum zone of inhibition was observed in 100g concentration of methanolic extracts of stem, berries and whole plant against to all bacterial strains. Similar antibacterial activity was showed by solanum species in different studies. Six different solvent extracts of Solanum nigrum leaf, seed and roots against to pathogenic bacteria such as B subtilis, B megaterium, Proteus vulgaris, Staphylococcus aureus, Klebsiella pneumonia and E.coli were also showed antibacterial activity, ethanolic extract of leaf showed highest activity than seed and root. (Sridhar et al., 2011). Patil Suhas et al. (2009) evaluated the antibacterial of Solanum surattense of methanolic extract of Whole plant using agar disc diffusion method, showed antibacterial activity against to Gram + ve bacteria (B subtilis and Streptococcus aureus) and it did not show considerable activity against to Gram -ve bacteria (E coli and Pseudomonas aeruginosa). In other studies, whole plant extract of Camellia sinensis (Yam e tal., 1997), extracts of stem, root, leaf, flower and whole plant of Tridax procumbens, (Aniel Kumar and Mutyala Naidu 2010), Andrographis paniculata (Aniel Kumar et al., 2010), Andrographis serpyllifolia (Revathi et al., 2012), Sida spinosa Linn. (Selvadurai et al., 2011), also showed antibacterial activity, among all extracts whole plant extract showed highest activity against to bacteria. The standard drug streptomycin (10g/ml) showed high degree of inhibition against E. coli and Klebsiella pneumoniae. CONCLUSIONS By the Sridhar et al. (2010) and present studies, antibacterial activity of all parts of the Solanum nigrum was evaluated. From the above results it can be concluded that Solanum nigrum extracts have great potential as antibacterial compounds against tested bacterial strains and that they can be used in the treatment of infectious diseases caused by resistant microorganisms. Antibacterial properties of Solanum nigrum plant extracts or whole plant extract can be effectively used for common infectious diseases causing bacterial species. ACKNOWLEDGEMENT We thank Dr. R. Senthil Kumar, Head, Department of Biotechnology, during the study and also for the provision of the media and other supplements required for the work. We also thank to Indian Academy group of institutions, Bangalore, Karnataka, for their full support to do work. REFERENCES
1.Aniel Kumar O. and L. Mutyala Naidu (2010). Antibacterial potential of Tridax procumbens L. against human pathogens. 2.Aniel Kumar O, Mutyala Naidu L and Raja Rao K.G. (2010). In vitro antibacterial activity in the extracts of Andrographis paniculata Burm. F. International Journal of PharmTech Research. 2: (2).1383-1385, 3.Bari ,M.A., Islam,1 W.,. Khan A.R . and Abul Mandal. (2010). Antibacterial and Antifungal Activity of Solanum torvum (Solanaceae). International Journal of Agricultural journal .12 (3): 386-390. 4.Haraguchi H, Kataoka S, Okamoto S, Hanafi Mand & Shibata K. (1999). Antimicrobial triterpenes from Ilex integra and the mechanism of antifungal action. Phytotherapia Residence, (13): 151-156. 5.Hashim H, Kamali EL, Mohammed Y. (2010). Antibacterial activity and phytochemical screening of ethanolic extracts obtained from selected Sudanese medicinal plants. Current Research Journal of Biological Science, 2(2): 143146. 6.Iwu, M.W., Duncan, A.R and Okunji, C.O. (1999). New antimicrobials of plant origin. In: Janick J.ed. Perspectives on New Crops and New Uses. Alexandria, VA: ASHS Press; pp. 457-462. 7.John Britto S, 2001. Comparative antibacterial activity study of Solanum incanum L. J. Swamy Botany Cl, 18: 8182. 8.John Britto S & Senthilkumar S, 2001. Antibacterial activity of Solanum incanum L. leaf extracts. Asian Journal of Microbiology Biotechnology Environmental Science, 3: 65-66. 9.Mahesh B. and Satish S. (2008). Antimicrobial ativity of some important medicinal plant against plant and human pathogens. World Journal of Agricultural Sciences 4 (S): 839-843. 10.Patil Suhas, Joshi Vijaya, Sutar Prasanna and Sambrekar Sudhir, (2009). Screening of whole plant extract of Solanum surattense for antibacterial activity. International Journal of pharmaceutical Sciences, 1(1):110-114. 11.Pavitra P.S. Janani V.S., Charumathi k.H., Indumathy R., Sirisha Potala and Rama S. Verma (2012). Antibacterial activity of plants used in indian herbal medicine. International journal of green Pharmacy, 23-28. 12.Revathi S.L., Suresh Kumar P., Sudarshana Deepa V, Senthil Kumar J and Anitha Janet Roshni.Y. (2012).

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In Vitro Antibacterial Activity In The Extracts Of Solanum Nigrum

Antimicrobial ial activity of Andrograpserpyllifolia folia (Rohl.ex.Vahl) Wrigh. International Journal of Pharmaceutical Science and Health care. 2: (1). 17-31. 13.Samy, R.P. and S. Ignacimuthu, (2000). Antibacterial activity of some folklore medicinal plants used by tribals in Western Ghats of India. J. Ethnopharmacol., 69: 63-71. 14.Sashikumar J M, RemyaM & Janardhanan K, 2003. Antimicrobial activity of ethno medicinal plants of Nilgiri biosphere reserve and Western Ghats. Asian Journal of Microbiology Biotechnology and Environmental Science, 5, 183-185. 15.Selvadurai S., Senthamarai R., Sri Vijaya Kirubha T. and Vasuki K. (2011). Anitimicrobial activity of ethanolic extract of the whole plant of Sida Spinosa Linn. (Malvaceae). Journal of Natural Product and Plant Resources. 1 (2): 3640. 16.Sheeba, E. (2010). Antibacterial activity of Solanum surattense Burm. F. Kathamandu University Journal of science, Engiineering and technology. 6: (I). 1-4. 17.Sridhar T.M., Josthna P. and Naidu C.V. (2011). In Vitro Antibacterial Activity and Phytochemical Analysis of Solanum nigrum (Linn.) - An Important Antiulcer Medicinal Plant. Journal of Experimental Sciences, 2(8): 24-29. 18. Werner, F., P. Okemo and R. Ansorg, (1999). Antibacterial activity of East African Medicinal plants. J. Ethnopharmacol., 60: 79-84. 19.World Health Organization. 2002. WHO Traditional medicine strategy 2002-2005, World Health Organization. 20.Yam T.S., Shah S., Hamilton- Miller J.M.T. (1997) Microbiological activity of whole and fractioned crude extracts of tea (Camellia sinensis), and of tea components. FEMS Microbiology Letters, 152: 169 -174.

Table 1: Antibacterial activity of ethanolic and methanolic extracts of Solanum nigrum.

Zone of Inhibition (mm) Ethanolic extracts Bacterial strains Concentration (g)

10

Methanolic extracts Whole plant

2..2 0.25 8.50.3 17.20.25 7..60.20 15.1 0..2 6.1 0.15 13.0 0.11 9..6 0.15 19.1 0.32

Stem
3..2 0.11 9.6 0.15 4.6 0.15 8.2 0.1 4..5 0.15 6.0 0..3 3..3 0.1 6..2 0.20

Berries
7..3 0.57 8.0 0.20 4.0 0.26 6.4 0.15 5.0 0.25 3.4 0.10 7.7 0.20 3.0 0.1 5.1 0.20

Stem
6..5 0.45 15.5 0.45 24.1 0.95 7..6 0.55 11.0 0.20 17.0 0.20 10.0 0.15 13.4 0.45 20.3 0.30 3.1 0.15 8..3 0.32 11.2 0.20

Berries
4..1 0.41 12.9 0.60 18.4 0.89 11.0 0.35 16.0 0.12 27.1 0.32 6..3 0.30 14.1 0.15 23.2 0.26 2.1 0.23 5..9 0.25 12.1 0.11

Whole plant
13..2 0.2 22.8 0.75 27.2 0.87 17.2 0.25 17.0 0.15 29.3 0..9 12.2 0..25 18.0 .0..25 24.7 0..32 4.8 0..20 10.2 0.25 14.3 0.30

Antibiotic (10 g)
10

Bacillus subtilus

50 100 10 50

16

E. coli

100 10

17

Klebsiella pneumonia

50 100 10

10

Pseudomonas aeruginosa

50 100

Values are mean S.D. of three readings.

Indian Streams Research Journal Volume 2 Issue 7 Aug 2012