Antibacterial Action of Medicinal Plant Alysicarpus Vaginalis Against Respiratory Tract Pathogens
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ESSENCE - International Journal for Environmental Rehabilitation and Conservation Volume VII: No. 2 2016 [25 – 32] [ISSN 0975 - 6272] [www.essence-journal.com] Antibacterial action of medicinal plant Alysicarpus Vaginalis against respiratory tract pathogens Prashant Arya1, J. P Mehta1 and Sanjay Kumar2 Received: July 28, 2016 Accepted: October 22, 2016 Online: December 31, 2016 Abstract Since the beginning of human civilization, aureus MTCC 1144, Streptococcus medicinal plants have been used by mankind pneumoniae MTCC 655, Streptococcus for its therapeutic value. Nature has been a pyogenes MTCC 442, Klebsiella pneumoniae prime source of medicinal agents for thousands MTCC 4030 and Pseudomonas aeruginosa of years and an impressive number of modern MTCC 2474. Plants root were collected from drugs have been isolated from natural sources. different parts of Garhwal Himalaya and The Himalayan natives have a great wealth of authenticated at Botanical Survey of India, medicinal plants and traditional medicinal Northern Regional Center Dehradun. Properly knowledge. Monitoring of susceptibility of washed with water jet and shade dried Root of herbal drugs for bacterial pathogens is the plant at room temprature.The plant material important for appropriate choice of treatment. were crushed and extracted according to The present study was to evaluate the polarity, low to high in petroleum ether (PT), antibacterial activity and phytochemical chloroform (CH), methanol (MH) and Water analysis of various extracts of root of A. (WT) by using Soxhlet apparatus. The vaginalis (Chukalai) against selected common antibacterial activity was examined by agar respiratory tract pathogens i.e. Staphylococcus well diffusion method and the minimum inhibitory concentration (MIC) by twofold Keywords: Antibacterial activity | Agar well serial dilution method. Erythromycin was used diffusion method | Twofold serial dilution as positive control to determine the sensitivity method | Phytomedicine | Respiratory tract of the strains and Di Methyl Sulphoxide pathogens | Alysicarpus vaginalis (DMSO) used as negative control. The results For correspondence: showed that methanolic extract was more 1 active than other extracts in its antibacterial Department of Botany and Microbiology, H.N.B. Garhwal University Srinagar (Garhwal), India 2Department of Botany and Microbiology, Gurukul Kangri activity. The zone of inhibition exhibited by University (Haridwar) Email- [email protected] methanol extract against tested micro- 25 Arya et al./Vol. VII [2] 2016/25 – 32 organisms ranged between 06 mm to 13 mm, vascular plants, 1748 spp. species are medicinal. respectively. MIC values were recorded Uttarakhand is a store house of a rich variety between 3.12 mg/ml to 25 mg/ml for all the herbs and medicinal and aromatic plant species. organisms. This investigation supports a good Medicinal plants produce a wide variety of response to the use of A.vaginalis in herbal compounds which in addition to give them medicine and as a base for the development of characteristic pigments, odour and flavor new drugs and phytomedicine in foundation characteristics may also have antimicrobial for its use in treatment of respiratory infectious properties (Cowan, 1999). For thousands of diseases. years, traditional plant derived medicines have Introduction been used in most parts of the world and their use Our earth is full of medicinal plants. These in fighting microbial disease is becoming the medicinal plants having therapeutical properties focus on study (Bhavnani & Ballow, 2000 are very useful in curing various diseases. Almost Chariandy et al., 1999). Intensive studies on all our present medicines exhaled from medicinal extracts and biologically-active compounds plants. Approximately, 8000 species of medicinal isolated from medicinal plants have played an plants are used as different systems of medicines essential role in drug discovery in last decade. in India (Planning Commission, 2000). India is Various parts of such plants like root, tubers, blessed with huge biodiversity due to different bark, flowers, leaves and seeds are used for climatic zones, in which numerous medicinal medicinal purposes. More so, many of these plants were reported. The Indian state of plants have been known to synthesize active Uttarakhand, located in central Himalayan region, secondary metabolites such as phenolic is richly gifted with a large variety of plant compound found in essential oils with established species, many of which have medicinal potent insecticidal (Kambu et al., 1982) and properties. Medicinal plants play an important antimicrobial activities, which really has formed role in the lives of people in Uttarakhand by the basis for their applications in some providing basic health care and employment to pharmaceuticals, alternative medicines and the farmers (Alam & Kop., 2005). The Central natural therapies (R´ıos & Recio, 2005; Reynolds Himalayan Region covers the new state of India, 1996; Lis-Balchin & Deans, 1997). As if a provides excellent opportunities for studying the comparable study is made between antibiotics Traditional Knowledge Systems (TKS). The and pathogen resistance it is concluded that Indian Himalayan region alone supports about antibiotics provide the main basis for the therapy 18,440 species of plants (Angiosperms: 8000 of microbial (bacterial and fungal) infections. spp., Gymnosperm: 44 spp., Pteridophytes: 600 But, overuse of antibiotics has become the major spp., Bryophytes: 1736 spp., Lichens: 1159 spp. factor for the emergence and dissemination of and Fungi: 6900 spp.) of which about 45% are multi-drug resistant strains of several groups of having medicinal properties. According to microorganisms (Harbottle et al., 2006). Thus, in Samant et al., (1998) out of the total species of the light of the evidence of rapid global spread of 26 Arya et al./Vol. VII [2] 2016/25 – 32 resistant clinical isolates, the need to find new material was washed jet properly, dried under antimicrobial agents is of paramount importance. shade at room temperature and crushed to Alysicarpus vaginalis small pieces by using pestle and motor. The genus Alysicarpus belongs to family Preparation of Extract - Plant extracts were Fabaceae. Commonly called as Chukalai, prepared by immersing 200g of powdered There are approximately 78 species in their plant material in 600 ml of four different genus with 20 species are reported in India solvents according to polarity low to high i.e. (Haines, 1978). They commonly found in open petroleum ether (PT), Chloroform(CH), grass land, crop fields, and way sides, altitude methanol (MH) and water (WT), loaded in 600 to 1000m amsl Srinagar, pouri, Chauras Soxhlet assembly and extracted for 72 h and Kirtinagar in Garhwal Himalaya, almost through successive method (Ahmed et al., throughout India, ascending to 1000m 1998). Plant extracts were filtered through Afghanistan, Pakistan and tropical America. Whatman No. 1 filter paper and crude extracts The genus comprises annual Prostrate Herbs, obtained by removing solvent in vacuum perennial. Glabrous or a line of hairs on stem, evaporator at 30°C. Residues were stored at erect or diffuse at 30cm long, branched Leaves, 4°C until further use. Extracts were dissolved flowers small, mostly ca.5 mm, usually binate in dimethyl sulfoxide (DMSO) to a final at each node of rachis (Gaur R D., 1999). concentration of 200mg/ml for agar well Several species of Alysicarpus has been used in diffusion method. indigenous system of medicine an anti- Test Microorganisms - The five bacterial inflammatory in stomachache, and also an strains causing respiratory infections used in antidote to snake bite. It is also used in skin this study were Klebsiella pneumoniae MTCC diseases and as a diuretic. The leaves are used 4030, Pseudomonas aeruginosa MTCC 2474, in fever, jaundice and leaf paste is applied Staphylococcus aureus MTCC 1144, externally on skin allergy (Shankarnarayan, Streptococcus pneumoniae MTCC 655, 1988). Chemical Constituents of A. vaginalis is Streptococcus pyogenes MTCC 442. These Caffeic acid, Catechol, Cumaric acid, Gentisic Bacterial strains were procured from Institute acid, Gallic acid, P-hydroxybenzoic acid of Microbial Technology (IMTECH), Syringic acid, Vanillic acid, Salicyclic acid, Chandigarh. Acetic Acid, Ethyl acetate. (Shahin and Preparation of Inoculums – Stock cultures Ahmad, 2012). were maintained at 4°C on slopes of nutrient Materials and Methods agar. Active cultures for experiment were Plant Material - Plant was collected from prepared by transferring a loopful of cells from Chauras Kirtinagar (Tehri Garhwal) stock cultures to test tubes of Mueller-Hinton Uttarakhand and authenticated at Botanical broth (MHB) for bacteria that were incubated Survey of India (BSI), Northern Regional without agitation for 24 h at 37°C. Center Dehradun. Collected plant Root 27 Arya et al./Vol. VII [2] 2016/25 – 32 Antibacterial testing – The antibacterial tube. Control tubes were inoculated with same activity of different extracts was determined by quantity of sterile distilled water. All tubes agar well-diffusion method (Ahmed et al., were incubated at 37°C for 24 h. The lowest 1998). 0.1 ml of 12-16 h incubated cultures of concentration that did not permit any visible bacterial species were mixed in molten Mueller growth when compared with control was Hinton Agar medium no. 173 (Hi media Pvt. considered as the minimum inhibitory Ltd., Mumbai, India) and poured in pre- concentration. The MICs was considered as the