ANTIMICROBIAL ACTIVITY OF ETHANOLIC EXTRACTS OF
EIGHT MEDICINAL PLANTS USED IN TRADITIONAL MEDICINE IN
INDIA
Mrunal Sene P1,Ajay Mandal2* and Sandeep Kumar Dey3
1,2
Department of Biotechnology Engineering, Acharya Institute of Technology, Bangalore-560107, India.
3
Department of Biotechnology, Presidency College, Bangalore-560024, India.
ABSTRACT
Ethanolic extracts of some Indian plant species used in folk medicine were investigated for their antimicrobial activities against bacterial species Staphylococcus aureus, Escherichia coli, Proteus vulgaris and Pseudomonas aeruginosa. Zones of inhibition were observed in disc
diffusion for antimicrobial investigation against Gram-positive and Gram negative pathogenic bacteria. The plants showed 80% of antimicrobial activity, with significant difference in activity between the different plants. The most antimicrobially active plants were Hyptis suaveolens and Cyperus rotundus, whereas, the least active plant was Couroupita guianensis. The ethanolic extracts showed activity against
both Gram positive and negative bacteria.
KEYWORDS: Antibacterial activity; Ethanolic extract; zone of inhibition.
1. INTRODUCTION
Use of medicinal plants as traditional medicine is one of the common practices in India due to their wide pharmacological activities. Traditional medicines are being used at the primary health care level by many developed and developing countries. More than 80% of world’s
population depends on plants for primary health care needs. Since ancient time in India, herbal medicines have been the basis of treatment and cure for various diseases physiological conditions in traditional methods practiced such as Ayurveda, Unani and Siddha. Herbal medicines have various therapeutic uses such as healing wounds, treating inflammations due
Volume 4, Issue 7, 1529-1534. Research Article ISSN 2277– 7105
Article Received on 07 May 2015,
Revised on 28 May 2015, Accepted on 19 June 2015
*Correspondence for
Author
Ajay Mandal
etc. In the constant effort to improve the efficacy and ethics of modern medical practice, researchers are increasingly turning their attention to folk medicine as a source of new drugs (Haslam et al., 1989). Nowadays, the development of resistance by a pathogen to many of the commonly used antibiotics provides an impetus for further attempts to search for new antimicrobial agents to combat infections and overcome the problems of resistance and side effects of the currently available antimicrobial agents. Hence this in vitro study was aimed at screening selected Indian medicinal plants for their antimicrobial activity, evaluating their potential use in treating dermatomucosal infections caused by bacteria and fungus.
2. MATERIALS AND METHODS 2.1. Plant collection
In the current work plant species commonly used in folk medicine in India for the treatment of dermatomucosal infections were selected (Table 1). Mature plants were collected from several sites in the Bangalore areas in India during the January -March of 2015. Aerial parts of each medicinal plant were collected, dried in the shade and ground into a powdered material using an appropriate seed mill. Voucher specimens of the plants were deposited at the Department of Biotechnology, Acharya Institute of technology, Bangalore, India.
2.2. Preparation of the extracts and fractions 2.2.1. Ethanolic extracts
Aliquots (100 g) of each dry powdered plant were soaked in 95% ethanol, petroleum ether, or ethyl acetate until complete exhaustion of the herb. The extracts were filtered using Whatman filter paper no.1, and the filtrates were then evaporated under reduced pressure and dried using a rotary evaporator at 60°C. Dried extracts were stored in labelled sterile screw capped bottles at 20°C.
2.3. Antimicrobial activity test 2.3.1. Microorganisms
The microorganisms used included: Staphylococcus aureus (MTCC 902), Escherichia coli (ATCC 25922), Proteus vulgaris (MTCC 744), Pseudomonas aeruginosa (MTCC 7083) 2.3.2. Screening for antimicrobial activities.
using an inoculum containing 106 bacterial cells spread on Muller-Hinton agar plates (1 ml inoculums/plate). The discs (diameter, 6 mm) were each impregnated with 50 ml of extract (10 mg/disc) at a concentration of 200 mg/ml and placed on the inoculated agar and incubated at 37°C for 24 h for bacteria. On each plate an appropriate reference antibiotic disc was applied depending on the test microorganisms (Jawetz et al., 1995). Ampicillin, penicillin-G and, gentamicin served as positive controls for E. coli, S. aureus, P. aeruginosa and P. vulgaris, Reference antibiotic discs contained 10 mg antibiotic/disc. Each test was carried out in triplicate.
Table 1: Ethnobotanic data of studied plants
Sl. No Name of the plant Parts used Uses
1
Cynarospermum asperrimum (common name : Hill
blepharis)
leaves Some species of Blepharis are used in fever, malaria and inflammation
2 Tithonia diversifolia (Local
name : Lam numitlei) leaves Flower-heads are used for wounds and bruises.
3 Hyptis suaveolens (Local
name : Tukmaan) leaves
Plant is stimulant, carminative,sudorific and lactagogue. Infusion is used in catarrhal conditions, uterus
affections and parasitical cutaneous diseases. Roots are chewed with betel nuts as a stomachic, and its decoction is used as an appetizer.
4 Melilotus indicus (Local name
: Ingkhol yensil) shoot
Plant is used as a discultient and emollient, externally is used as a fomentation, poultice or plaster for swellings. The seeds are useful in bowel complaints and infantile diarrhea.
5 Couroupita guianensis Aubl (Cannon Ball Tree)
flower and bark
anti-inflammatory , skin infections, boils, malaria, stomachache and sores, larvicidal activity
6 Phyllanthus emblica(Indian
Gooseberry) fruits Sinusitis, vomiting, anaemia ascities, piles 7 Cyperus rotundus (nut
Grass) leaves Tuberculosis,fevetr,sweating,vomiting,dysentry 8 Terminalia chebula(hebulic
Myrobalan) bark Ulcer,asthma,ascities, brain tumor,fistula
Table 2: Antimicrobial activity of plant extracts against bacteria.
Sl. No Ethanolic extracts Inhibition zone diameter (mm)
S. aureus E. coli P. aeruginosa P. vulgaris
1 Tithonia diversifolia 6.2 6.2 5.8 7.2
2 Hyptis suaveolens 14.3 6.0 6.0 10.1
3 Melilotus indicus 7.0 4.8 5.0 5.6
4 Couroupita guianensis 5.0 5.0 5.0 5.6
5 Phyllanthus emblica 8.1 6.0 5.9 6.3
6 Cyperus rotundus 14.2 10.2 7.2 11.0
2.4. Statistical analysis
Data were analyzed and treatments compared using the 3-way ANOVA with 95% confidence limits (PB0.05).
3. RESULTS AND DISCUSSION 3.1. Antimicrobial activity of 8 plants
Antimicrobial activity of ethanolic extracts of 8 plants (Table 1) has been evaluated in vitro bacterial species which are known to cause dermic and mucosal infections. All plants studied in this work showed antimicrobial activity against some of the test microorganisms. This result showed that most of the studied plants (90%) are potentially a rich source of antimicrobial agents. However, the plants differ significantly in their activity against test microorganisms. The most active plants were Hyptis suaveolens and Cyperus rotundus, whereas, the least active plant was Couroupita guianensis (Table 2). The ethanolic extract of Tithonia diversifolia was effective against P. vulgaris, E. coli and S. aureus showing inhibition zones of 7.2, 6.2 and 6.2 mm, respectively. The ethanolic extract of Hyptis suaveolens was effective against S. aureus and P. vulgaris showing inhibition zones of 14.3 and 10.1 mm, respectively.The ethanolic extract of Melilotus indicus was effective against S. aureus showing inhibition zones of 7.0 mm, respectively. The ethanolic extract of Couroupita
guianensis was effective against P. vulgaris, showing inhibition zones of 5.6 mm, respectively. The ethanolic extract of Phyllanthus emblica was effective against S. aureus showing inhibition zones of 8.1 mm, respectively. The ethanolic extract of Cyperus rotundus was effective against S. aureus and P. vulgaris, showing inhibition zones of 14.2 and 11.0 mm, respectively. The ethanolic extract of Terminalia chebula was effective against S. aureus showing inhibition zones of 7.5 mm, respectively. The ethanolic extract of
Cynarospermum asperrimum was effective against S. aureus showing inhibition zones of 6.1
mm, respectively.
and demonstrates the importance of such plants in medicine and in assisting primary health care in this part of the world.
REFERENCES
1. Dafni, Yanive, Z., Solanaceae as medicinal plants in Israel, Journal of Ethnopharmacology, 1994; 44: 11±18.
2. Haslam, E., Lilley, T.H., YaCai, Martin, R., Magnolato, D., Traditional herbal medicines the role of polyphenols, Planta Medica, 1989; 55: 1±8.
3. Hussain, A.I.A., 1995. Isolation, identi®cation and determina-tion of antifungal constituents from some medicinal plants in Palestine. MSc Thesis, An-Najah National University, Nablus, Palestine.
4. Jawetz, E., Melnick, J.L., Adelberg, E.A., Brooks, G.F., Batel, J.S., Ornston, L.N., 1995. Medical Microbiology, vol. 20th ed. Appleton and Lang, New York.
5. Kandil, O., Radwan, N.M., Hassan, A.B., Amer, A.M.M., El-Banna, H.A., Extracts and fractions of Thymus capitatus exhibit antimicrobial activities. Journal of Ethnopharmacology, 1994; 44: 19±24.
6. Medoff, G., Kayashi, G.S., Mode of action of antifungal drugs. In: Howard, D.H., Howard, L.E. (Eds.), Fungi Pathogenic for Humans and Animals. Part B, Pathogenic-ity and Detection: 1. Marcel Dekker, New York: 1993, pp. 325±349.
7. Murray, P.R., Baron, E.J., Pfaller, M.A., Tenover, F.C., Yolke, R.H., 1995. Manual of Clinical Microbiology, vol. 6th ed. ASM, Washington, DC.
8. Palevitch, P.D., Yaniv, Z., 1991. Medicinal Plants of Holly-land, vols. 1±2. Tammuz, Tel-Aviv.
9. Rojas, A., Hernandez, L., Pereda-Miranda, R., Mata, R., Screening for antimicrobial activity of crude drug extracts and pure natural products from Mexican medici-nal plants. Journal of Ethnopharmacology, 1992; 35: 275±283.
10.Shtayeh, M.S., Hamad, A.Kh., 1995. Protection of the Palestinian Environment (in Arabic), Nablus.
11.Silva, F., Abraham, A., The potentiality of the Israeli ¯ora for medicinal purposes. Fitoterapia, 1981; 52: 195±200.
12.Yaghmour, R., 1997. Antimicrobial activity of 20 plants used in folkloric medicine in Palestine. MSc Thesis, An-Najah National University, Nablus.
