International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202Vol. 2(6), 60-62, June (2013) Int. Res. J. Biological Sci. International Science Congress Association 60 Short CommunicationEvaluation of Antibacterial Activity of Tecomella undulata leaves crude ExtractsSharma Abhishek1*, Patil Ujwala, KakkarShivani and Bhot MeetaDepartment of Botany, Birla College, Kalyan- 421304, MS, INDIA Department of Biotechnology, Birla College, Kalyan- 421304, MS, INDIAAvailable online at: www.isca.in Received 25th March 2013, revised 30th April 2013, accepted 3rd May 2013Abstract In the present study antibacterial activity of Tecomella undulata, leaves were checked on bacterial pathogens of humans. Hexane, chloroform and methanol extracts were used for the study by agar well diffusion method against seven bacteria. The methanol extract showed higher antibacterial activity in comparison to the other two solvents. Methanolic extract showed higher minimum inhibitory concentration against Klebsiella pneumoniae and Micrococcus luteus; whereas, less inhibitory effect was noted for chloroform and hexane extracts. Gram-negative bacteria were more susceptible than Gram-positive. Present study validates the traditional use of Tecomella undulata in medicine. Keywords: Tecomella undulata, Antibacterial activity, Agar-well diffusion. Introduction Tecomella undulata (Family, Bignoniaceae) is commonly known as Ammora (in English) or Rohida and is locally known as honey tree, desert teak, Marwar teak or white cedar. It is widely distributed in Arabia, southern Pakistan and northwest India up to an elevation of 1200 meters. In Pakistan, it is found in Baluchistan and Sind. Iridoid Glucoside, naphthoquinone, phytosterols, flavonoid glycoside, flavonol, fatty acid and triterpenoids were identified from different parts of Tecomella undulata which are pharmacologically active. It is reputed to be of considerable medicinal value in Ayurveda and traditionally, it is extensively used for treatment of several diseases. As no data on anti bacterial activity is available on Tecomella undulata, present study was proposed to check antibacterial potential of Tecomella undulata against Bacillus subtilis, Enterococcus faecalis, Escherichia coli, Klebsiella pneumonia, Micrococcus luteus, Proteus vulgaris and Pseudomonas aeruginosa using hexane, chloroform and methanol leaf extracts. Material and Methods The present study was done in the Department of Botany, Birla College, Kalyan. Leaves of Tecomella undulata were collected from Kalyan during winter season. Source of Microorganisms: Pure bacterial cultures were obtained from NCIM, Pune, India and were maintained in nutrient agar (Himedia)table 1. Extraction procedure: Leaf was washed under running tap water, excess water was drained off, shade dried and finally crushed to fine powder in mixture grinder. The dried powder of the leaves was allowed to Soxhlet for successive extraction with hexane, chloroform and methanol. The liquid extract were collected, filtered and evaporated to dryness. The extracts were dissolved in respective solvents to reach a low and high concentration (100 mg/ml and 300 mg/ml) and were kept in refrigerator till used. Table-1 List of tested bacterial strains and their catalogue numbers Bacteria Catalogue number Bacillus subtilis 2545 Enterococcus faecalis 2356 Escherichia coli 2641 Klebsiella pneumoniae 2883 Micrococcus luteus 2704 Proteus vulgaris 2813 Pseudomonas aeruginosa 2026 Antibacterial activity: The antibacterial activity was carried out by agar well diffusion method2-4. For this, the different solvent extracts of leaf of Tecomella undulata have been tested against gram positive and gram negative bacteria to find out the antibacterial activity. A nutrient broth was prepared (beef extract-3gm/L; Peptone-5gm/L; NaCl-5gm/L; Agar-16gm/L) and then sterilized in an autoclave at 15lb pressure for 15 minutes. The sterilized medium was poured into petri dishes. The solidified plates were bored with 6mm diameter sterilized cork borer. The plates with wells were used for the antibacterial studies. Various solvent extracts of leaf of Tecomella undulatawere subjected to individual microbiological tests to ascertain their antibacterial activity against seven species of microorganisms, Bacillus subtilis, Enterococcus faecalis, International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(6), 60-62, June (2013) Int. Res. J. Biological Sci. International Science Congress Association 61 Escherichia coli, Klebsiella pneumonia, Micrococcus luteus, Proteus vulgaris and Pseudomonas aeruginosa. The evaluation of antibacterial activity of the extracts was done by observing the clear zone around disks and measuring the diameter of the zone of inhibition (ZI) exhibited by the extracts around disks in mm by sliding digital vernier caliper. The Minimum inhibitory concentration (MIC) was determined through the dilution method5-10. Different concentrations of leaf extract in hexane, chloroform and methanol (ranging from 0.01 to 5.00 mg/ml) was tested separately for each bacterium and inhibition zone of microbial growth in the plates containing test solutions was judged by comparison with blank control plates. Experiments were performed in triplicate. The results of MIC were interpreted as sensitive (14 mm), intermediate activity (14-17 mm) and susceptible (&#x-3.3;堦17 mm). Results and DiscussionThe results of antibacterial activity and MIC values of solvent extracts of Tecomella undulata leaves are summarized in tables 2 and 3, respectively. Methanolic extract was found to be more potent as compare to n-Hexane and chloroform extracts against all tested bacteria. Hexane extract was found to be resistance against Klebsiella pneumoniae, Pseudomonas aeruginosa and Micrococcus luteus for low concentration, while Klebsiella pneumoniae showed intermediate activity to its high concentration. Escherichia coli, Proteus vulgaris, Bacillus subtilis and Enterococcus faecalis did not show any inhibition zones. Chloroform extract was found effective against Klebsiella pneumoniae and Micrococcus luteus showed intermediate activity against Pseudomonas aeruginosairrespective of its concentrations whereas Escherichia coli, Proteus vulgaris and Enterococcus faecalis were resistance to low concentration and evinced intermediate activity to high concentration. Bacillus subtilis did not show inhibition zone. Methanol extract exhibited promising activity against all tested bacteria for both concentrations. It showed fairly high degree of sensitivity against Klebsiella pneumoniae followed by Micrococcus luteus. Compared with standard antibiotic, methanol extract showed the high inhibition zone values than ciprofloxacin (10g). Hence it indicated that methanol extract of Tecomella undulate leaves has broad spectrum of antibacterial activity against all tested bacteria. Hexane, methanol and chloroform, a negative control, did not show zone of inhibition and indicated that it is not interfering with the formation of zone of inhibition. From the MIC values, it is observed that methanol extract showed significant antibacterial activity followed by chloroform and hexane extracts table 3. The MIC value of methanol extract was 0.01 mg/ml for Klebsiella pneumoniae while 0.1 mg/ml against Micrococcus luteus and 1.0 mg/ml for Enterococcus faecalis. The MIC value of chloroform extract for K. pneumoniae was 1.0 mg/ml, whereas for M. luteus it was 3.0mg/ml. The hexane extract exhibited the same value against all tested bacteria, it was &#x-3.3;堦5.0 mg/ml. Table2 Inhibitory zones of solvent extracts of Tecomella undulata against Gram-negative and Gram-positive bacteria Solvent Extract Zone of inhibition (mm) Concentrations (mg/ml) BS EF EC KP ML PV PA Hexane 100 Nil Nil Nil 12 11 Nil Nil 300 Nil Nil Nil 14 13 Nil 12 Chloroform 100 Nil Nil 12 13 19 13 15 300 16 13 14 23 20 15 17 Methanol 100 18 16 19 25 23 19 20 300 17 20 21 27 25 21 22 Ciprofloxacin 10g Nil 23 15 20 22 21 20 Negative Control Nil Nil Nil Nil Nil Nil Nil Table-3 MIC value of Tecomella undulata leaf extracts Bacteria MIC values (mg/ml) Methanol extract Chloroform extract Hexane extract Bacillus subtilis4.0 �5.0 �5.0 Enterococcus faecalis 2.0 �5.0 �5.0 Escherichia coli 1.0 �5.0 �5.0 Klebsiella pneumonia 0.01 1.0 �5.0 Micrococcus luteus 0.1 3.0 �5.0 Proteus vulgaris 2.0 5.0 �5.0 Pseudomonas aeruginosa 2.0 4.0 �5.0 International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(6), 60-62, June (2013) Int. Res. J. Biological Sci. International Science Congress Association 62 In the present study hexane, chloroform and methanol extracts of leaves were found to be the most sensitive against K. pneumoniae and Micrococcus luteus strains. Among the three extracts, the methanol extract exhibited promising results than chloroform and hexane extracts. Methanol proved as the most effective solvent for extracting broad spectrum of antimicrobial compounds from plants. Conclusion In conclusion, the present investigation of antibacterial activity of leaf extracts of Tecomella undulata reports that the various solvent extracts of leaf showed the antibacterial activity against bacterial pathogens of both gram positive and gram negative. Thus it can be employed as an antibacterial agent against many bacterial diseases. Further its, chemical characterization will disclose the bioactive phytoconstituents responsible for inhibitory activity of Tecomella undulata. References 1.Jain M., Kapadia R., Jadeja R. 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