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In the present study the antimicrobial activity of chloroform and methanolic extracts of Opuntia cochenillifera for both cladode and fruits was investigated. Methanolic extract was found to be an effective against the microbes namely, E. coli, Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa Candida albican C.glabrata C.haemulonii C.Tropicalis. Maximum activity was observed against E. coli, B. Subtilis and, C.albican and C.glabrata at 40mg/ml. Agar well diffusion assay was used to determine minimum inhibitory concentration of all test microorganisms. Introduction Nowadays, antimicrobial resistance has become a major global problem which is caused by the indiscriminate use of antimicrobial drugs. The use of medicinal plants, which provide a rich source of novel antimicrobial agents in human history as many infectious diseases have traditionally been treated with herbal medicines. Plant based antimicrobials represent a vast source for medicines and further exploration of the plant. Antimicrobials of origin have enormous therapeutic potential [1]. Human infections, especially those involving microorganisms i.e. bacteria, fungi, cause serious infections in tropical and subtropical countries of the world. In recent time, multiple drug resistance in human pathogenic microorganisms has been developed due to indiscriminate use of commercial antimicrobial drugs commonly used of such diseases [2, 3]. In general, bacteria have the genetic ability to transmit and acquire resistance to drugs which are utilized as therapeutic agents [4]. Medicinal plants represent a rich source of antimicrobial agents. Plants are used medicinally in different countries and are a source of many potent and powerful drugs. A wide range of medicinal plant parts are used to extract as raw drugs which possess different medicinal properties. Medicinal plants are major source of new chemical substances with potential therapeutic effects [5]. There is need to identify novel substance active towards highly resistant pathogens until natural products have been approved as new antimicrobial drugs, [6, 7]. Opuntia species have been used by humans for thousands of years [8] besides being consumed as food or beverages. Most portions of the plants have been used as medicine. This species is free of spines and spine-hairs on maximum portion [9, 10]. This plant has been used to treat rheumatism, diarrhoea and other inflammatory problems as well as a diuretic and analgesic, particularly for ear and tooth aches [11, 15]. The anti-inflammatory principle was related to a b-sitosterol [16]. Despite these biological properties, validation of any antibiotic potential against bacterial and fungal species has not been reported much from India. International Journal of Pharmacology, Phytochemistry and Ethnomedicine Submitted: 2016-06-16 ISSN: 2297-6922, Vol. 3, pp 84-89 Accepted: 2016-06-24 doi:10.18052/www.scipress.com/IJPPE.3.84 Online: 2016-06-30 © 2016 SciPress Ltd., Switzerland SciPress applies the CC-BY 4.0 license to works we publish: https://creativecommons.org/licenses/by/4.0/ Materials and Methods Plant material Habit Inflorescence Fruit Collection of raw materials The plant material collected from Gulbarga University campus located at southern part of India. The Plant species were identified with the reference of the Digital flora of Karnataka, Flora of Presidency of Madras, Flora of Gulbarga district and the Flora of Karnataka. [17, 19] The cladodes and fruits were shade dried, powdered mechanically and stored in airtight containers for extraction. Preparation of crude extracts The powdered material of opuntia cochenillifera fruit and cladode were subjected to successive solvent extraction. The powder is taken separately in 1 litre capacity thimble of soxlet apparatus and refluxed successively with chloroform and methanol for 48hrs in 8 batches of 500g each. Then it was allowed to evaporate which yield crude extracts used for antimicrobial activity. Test Microorganisms Authentic pure cultures of human pathogenic bacteria were obtained from the Medicinal plants and Microbiology laboratory, Department of Botany, Gulbarga University Kalaburagi Karnataka, India. And candida sps were taken from (MTCC) Chandigarh India Preparation of sample In the study of antimicrobial activity, the extract was dissolved in Dimethyl sulphoxide (DMSO). The corresponding concentration was expressed in term of mg of extract per ml of solvent (mg/ml). Antibacterial activity Antibacterial study (plate hole diffusion or agar well diffusion) assay was used to determine the growth inhibition of bacteria by plant extracts. Total of 25ml of nutrient agar plate was prepared and poured into sterile Petri dishes. The depth of the medium was approximately 4 mm. After the medium got solidified, the plates were allowed to dry for one hour. The bacterial/yeast suspension equal 1.5×108 cells /ml in sterile normal saline was prepared following the method [20] and inoculated on nutrient agar media by sterile cotton swabs. Wells in 6 mm diameter were punctured in the media using sterile cork borers. These wells were placed in Petri dishes allowing a distance of 2 to 4 cm between each well and filled with 20 μl of the different concentration extract .The plates were then incubated at 37 0 C for 24 hours. Following incubation, bioactivity was determined by measuring the inhibition zones around the crude extract in mm. All tests were done in triplicate. DMSO solvent is considered as negative control. Streptomycin was used as a standard antibacterial drug. International Journal of Pharmacology, Phytochemistry and Ethnomedicine Vol. 3 85 |
