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Antibacterial Potency Screening of the Crude Extracts of Adenanthera Pavonina L

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Antibacterial Potency Screening of the Crude Extracts of Adenanthera Pavonina L Universal Journal of Microbiology Research 3(3): 36-40, 2015 http://www.hrpub.org DOI: 10.13189/ujmr.2015.030302 Antibacterial Potency Screening of the Crude Extracts of Adenanthera pavonina L. Abdul Matin, W Islam*, O Ali Mandal Institute of Biological Sciences, University of Rajshahi, Bangladesh Copyright © 2015 by authors, all rights reserved. Authors agree that this article remains permanently open access under the terms of the Creative Commons Attribution License 4.0 International License Abstract The anitbacterial potency screening of the crude of A. pavonina have been used for the treatment of different extracts of leaves, seed and stem wood of Adenanthera human disorders like chronic rheumatism, diarrhea, pavonina L. were tasted against 15 Gram-positive and dysentery, antiseptic paste, pulmonary affections, chronic Gram-negative bacteria along with a standard antibiotic, ophthalmia and reported to have properties such as Ciprofloxacin. The in vitro screening for antimicrobial antioxidant, antibacterial by Jayasinghe et al. [2]. There have potency was carried out by following the agar diffusion and also been reports on cytotoxicity of crude extract determined micro broth dilution techniques. Among the fifteen bacterial using the brine shrimp assay observed by Ajaiyeoba et al. [3]. strains tested the chloroform fraction of the seed extract The plants also contain alkaloid, octacosanol, saponin, showed promising antibacterial efficacy against stigmasterol, HCN-glocoside, lignoceric acid and Salmonella-β-haemolyticus, Serratia dysenteriae and 2,4-di-OH-benzoic acid. Here we report a comparative Escherarichia coli. The susceptibility order of extracts were potency of extracts obtained in different solvents from leaves, seed>leaf>stem. The MIC values of the chloroform seed seed and stem wood of A. pavonina to inhibit Gram-positive extract were determined 256µg/ml againstB. cerus; 128 and Gram-negative bacteria due to their random use in µg/ml against S.-β–haemolyticus, S. dysenteriaeand 64 customary and traditional medicine to cure common ailments µg/ml against Klebsiella sp. The MIC values of the such as rheumatism, diarrhea, chronic ophthalmia etc. chloroform extracts of stem wood were128µg/ml S.-β–haemolyticus; 64 µg/ml against B. megaterium, S. sonnei and 32 µg/mlagainst S. typhi. 2. Materials and Methods Keywords Adenanthera Pavonina, Extract, Plant collection and identification: The plant specimen Antibacterial Screening, Disc Diffusion Assay, Zone of was collected from Botanical Garden, Rajshahi University Inhibition, MIC Campus. Identification of voucher specimens was confirmed at the Taxonomical Section, Department of Botany, University of Rajshahi, Bangladesh. Preparation of extract: The leaves, seeds and steam 1. Introduction woods was cut into small pieces, dried in shade, stored in cotton bags and then finely powdered (100 g) separately with The plants are the natural chemical factories that the help of a grinder. Each ground material was soaked in synthesize unmemorable compounds. The plants-derived 500 mL petroleum ether, acetone, chloroform and methanol compounds have been utilized by the human kind from time (BDH, pooleg England) separately for 24-72 h and filtered immemorial in different sectors of life, including public (Whatman No. 1). The filtered one was then allowed to health and pest management. The test plant is native to vaporize in rotary evaporator until completely dried and kept 0 Bangladesh and is easily available. They are well known for in a refrigerator at – 20 C with proper labeling. Then dried their medicinal values. extract (100 mg and 50 mg), for further study, was weighed Adenanthera pavonina L. is a deciduous, erect, unarmed and dissolved in 10 mL of respective solvents for dilution. tree 6-15 m high, distributed up to native to India, China, Antibacterial screening: Inhibitory activity of the Malaysia and Africa. In Bangladesh it is occasionally grown extracts was performed by following the agar well diffusion under cultivation reported by Zarnowski et al. [1]. It is a method of Perez et al. [4]. Fifteen pathogenic bacteria (six monogenic plant in family Fabaceae and commonly known gram-positive and nine gram-negative) were selected for the as Rakta chandan, Ranjana, Rakta kambal, Bead tree, Coral antibacterial test and were cultured at the Molecular wood tree, Red sandalwood, etc. Tradinally, different parts Laboratory, Institute of Biological Sciences, Rajshahi Universal Journal of Microbiology Research 3(3): 36-40, 2015 37 University. The test extracts were dissolved in respective amount (2.048 mg), and then broth medium (2 ml) was added solvents in such a manner that the desired concentrations (50 to each of the vials and agitated well to make sample solution and 200 μg/disc) for application in the disc have been whose concentration become 1024 µg/ml. The standard obtained. Standard antibiotic discs of Ciprofloxacin (30 antibiotic Ciprofloxacin solution 512 µg/ml was used for μg/disc) was also used for comparison. comparison. The six gram positive bacterial strains, viz., Staphylococcus aureus (ATCC-259233), Sercinia lutea (QL-166),Bacillus subtilis (QL-40),Bacillus megaterium 3. Results and Discussion (QL-38), Bacillus cereus (ATCC-14603) and Antibacterial Effect Streptococcus-β–haemolyticus (ATCC-10389) and nine gram negative bacterial strains, viz., Salmonella typhi The anitbacterial activity of A. pavonina extractives (ATCC-14028),Shigella dysenteriae (AL-35587), collected in petroleum ether, acetone, chloroform and Escherichia coli (FPFC-1407), Shigella shiga methanol of leaves, seed and stem wood were tested against (ATCC-26107), Shigella boydii (AL-17313), Shigella 15 bacteria. A correlation was found between the sonnei (AJ-8992), Proteus vulgaris, Klebsiella pneumoniae antibacterial activity observed by agar disc diffusion assay and Pseudomonus aeruginosa (ATCC-27853) were and MIC determination. obtained from the mother stock of the Enteric Microbiology The highest average zone of inhibition was found 25 mm Laboratory, ICDDR,B Dhaka and the Molecular Biology and 22 mm against: S.-β –haemolyticus by chloroform Laboratory, Institute of Biological Sciences, University of fraction of seed extract and stem wood extract and also Rajshahi, Bangladesh. against methanol fraction of same bacteria (Table 2) when Antibacterial activity was determined as diameter of compared to the ciprofloxacin 33 mm in diameter against inhibition zone using disc diffusion method of Perez et al. [4]. Proteus sp. of seed extract and S. typhi of stem wood extracts. Nutrient agar (NA) and Saboroud Dextrose agar (SDA) were Then moderate zone of inhibition were recorded respectively distributed in sterilized petridishes. This was accomplished against gram positive and gram negative bacterial strains in by placing 10µl of the extract on a small (6mm) filter paper both type of extraction either chloroform and methanol by disc. This disc was placed on an agar growth medium seed, leaf and stem wood presented in Tables (1-3). The containing a confluent lawn of microorganism. The highest average zone of inhibition was found 24 mm against concentration of the organism was also10µl/petridish. The S. boydii by pet. Ether and acetone fraction of leaf extracts absence of bacterial and fungal growth around the disc compared to the ciprofloxacin 33 mm in diameter against S. indicated that the plant extract contains antimicrobial typhi and Proteus sp. of seed extract and leaf extracts properties against that particular organism. respectively (Tables 1-2). Remaining of the lowest average Determination of Minimum Inhibitory Concentration zone inhibition activity was 6 mm against S. dysenteriae of (MIC): The serial dilution technique was followed using leaf extract of chloroform fraction whereas the comparative nutrient broth medium of Reiner [5] to determine the MIC study of ciproflozacin showed; 26 mm in diameter against values of the chloroform and methanol extracts against B. gram positive bacteria S. aureus and 28 mm in diameter cerus, B. Megaterium, S. aureus, S. sonnei and S. dysenteriae. against gram negative bacteria B. cereus, Klebsiella sp. The selected extracts were taken into different vials in a fixed Table 1. Antibacterial activity of the leaf extracts of A. pavonina and the standard Ciprofloxacin. Test organisms Diameter of zone of inhibition (in mm) each disc=6mm Chloroform extract Methanol extract Pet. Ether extract Acetone extract Ciprofloxacin 30 µg/disc 50 200 50 200 50 200 50 200 µg/disc Gram positive bacteria S. aureus 8 16 - - - - - - 33 B. cereus - - 9 20 9 19 10 20 28 B. megaterium 9 15 10 22 10 21 12 24 31 B. subtilis - - - - - - - - 30 S. lutea - - - - - - - - 31 S.-β-haemolyticus 10 20 8 17 11 23 - - 30 Gram negative bacteria S. typhi 8 15 7 16 - - - - 31 S. dysenteriae 6 17 - - 9 17 8 18 30 S. shiga - - - - 8 18 9 20 31 S. sonnei - - - - - - - - 30 S. boydii - - - - 12 24 10 21 30 E. coli - - - - - - - - 31 Klebsiella sp. 8 18 6 14 9 19 - - 28 P. aeruginosa - - - - - - - - 30 Proteus sp. - - - - - - - - 31 38 Antibacterial Potency Screening of the Crude Extracts of Adenanthera pavonina L. Table 2. Antibacterial activity of the seed extracts of A. pavonina and the standard Ciprofloxacin. Test organisms Diameter of zone of inhibition (in mm) each disc=6mm Chloroform extract Methanol extract Pet. Ether extract Acetone extract Ciprofloxacin µg/disc 50 200 50 200 50 200 50 200 30 µg/disc Gram positive bacteria S. aureus - - - - - - - - 26 B. cereus - 14 - 18 11 21 7 16 28 B. megaterium - 15 - 16 10 20 6 15 30 B. subtilis - - 10 20 - - - - 32 S. lutea - - - - 12 22 10 21 26 S.-β-haemolyticus 12 25 10 21 9 18 - - 32 Gram negative bacteria S. typhi - - - - 10 20 - - 33 S. dysenteriae 10 18 - - 11 21 8 16 29 S. shiga - - - - - - - - 30 S. sonnei - - - - - - - - 30 S. boydii - - - - 9 16 11 18 29 E. coli - 22 - - - - - - 29 Klebsiella sp. - - 9 16 12 20 10 19 31 P. aeruginosa - - - - - - - - 31 Proteus sp. - - - - - - - - 33 Table 3.Antibacterial activity of the stem wood extracts of A. pavonina and the standard Ciprofloxacin. Test organisms Diameter of zone of inhibition (in mm) each disc=6mm Chloroform extract Methanol extract Pet.
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