Comparative Antimicrobial Studies of Methanolic Extract of Muntingia calabura, Basella alba and Basella rubra Leaves.

K.B. Premakumari*, Ayesha Siddiqua, Shanaz Banu, J. Josephine, Leno Jenita and Bincy Raj

Dayananda Sagar College of Pharmacy, Kumaraswamy Lay Out, Bangalore.

ABSTRACT:

In vitro antimicrobial activity of methanolic extract of Muntingia calabura, Basella alba and Basella rubra leaves was investigated. The extracts exhibited marked antimicrobial activity against gram positive and gram negative bacteria and fungi. Muntingia calabura showed higher inhibitory effect against the pathogens like Staphylococcus aureus, Bacillus subtilis, Escherichia coli. Basella rubra showed mild inhibitory activity against Staphylococcus aureus, Basella alba showed good inhibitory activity against Aspergillus niger. A Comparative study shows maximum antimicrobial activity in Muntingia calabura extract.

KEYWORDS: Muntingia calabura, Basella alba and Basella rubra and antimicrobial activity.

INTRODUCTION:

Herbal medicine is the most ancient form of health care known to human kind. The word herb is used in herbal medicine (also known as botanical medicine or phytotherapy or phytomedicine) means a plant or plant part that is used to make medicines, food flavors or aromatic oils for soaps and fragrances. An herb can be a part of plant used for its medicinal property^1-3. Muntingia calabura, also known as ‘gasagase hanninamara’ in kannada, is c to the family Elaeocarpaceae⁴, The leaves possess antinociceptive, anti-inflammatory, antipyretic, antiulcer and antimicrobial activities and reduce swelling of prostate gland respectively⁵. The leaf infusion is drunk as a tea like beverages⁴. According to traditional use the flowers are said to possess antiseptic property and also antispasmodic activity. Therefore the present study was planned to evaluate the antimicrobial activity of the leaves of Muntingia calabura, Bascilla Alba and Bascilla Rubra The methanolic extract of Muntingia calabura leaves showed high amount of phenolic content and it is having antioxidant activity⁶.

Bascilla Alba and Bascilla Rubra belonging to the family Bascillacae which is commonly known as Malabar spinach. A number of medicinal applications have been reported. Young leaves are used as a laxative and the red fruits juices as eye drops to treat conjunctivitis. In Kenya leaves are used to cure stomachache and constipation after child birth. Pulped leaves are applied as a poultice and sores. Its fruits have been used for dyeing and also used as ink, cosmetics and for coloring food. The main chemical constituent reported in the literature Basella saponins [A, B, C and D]^7,8.

TABLE NO 1: Antimicrobial activity of the leaves of Basella alba, Muntingia calabura and Basella rubra against pathogenic species by cup plate method

SI. NO	SAMPLE	Zone of inhibition in mm
		Staphylococcus aureus		Bacillus subtilis		E .coli		Klebsiella pneumoniae		Aspergillus niger
		200 (μg/ ml)	100 (μg/ ml)	200 (μg/ ml)	100 (μg/ ml)	200 (μg/ ml)	100 (μg/ ml)	200 (μg/ ml)	100 (μg/ ml)	200 (μg/ ml)	100 (μg/ ml)
1	Basella rubra	11	Nil	Nil	Nil	Nil	Nil	Nil	Nil	Nil	Nil
2	Muntingia calabura	14	10	14	12	10	Nil	Nil	Nil	Nil	Nil
3	Basella alba	Nil	Nil	Nil	Nil	Nil	Nil	Nil	Nil	16	12
Standard drugs	Streptomycin (100 μg/ ml)	18	16	22	17	20	18	21	18	-	-
	Flucanazole (25 μg/ ml)									19	16

MATERIALS AND METHODS:

Collection of plant:

The leaves of Muntingia calabura, Basella alba and Basella rubra were collected and shade dried for about 2 weeks. The leaves are then coarsely powdered and were subjected to extraction process.

Collection of Organism:

The organisms were collected from Department of Microbiology, Dayananda sagar College of pharmacy, Bangalore.

Preparation of plant extract:

The leaves are then coarsely powdered and extraction is carried out by kinetic maceration process using methanol as solvent for one week. Methanolic extracts were filtered through muslin cloth and Whatman filter paper. The filtrate was concentrated and gummy extract was obtained.

Microorganisms:⁹

Pathogenic bacteria strains like, Staphylococcus aureus (NCIM 2079) (Gram positive), subtilis (NCIM 2063) (Gram positive), Eschericia coli (NCIM 2065) (Gram negative), Klebsiella pneumoniae (NCIM 2957) (Gram negative) and fungi Aspergillus niger (NCIM 2316) were used for the screening of antimicrobial activity.

Evaluation of Antimicrobial Activity:

The antimicrobial activity of the extracts was evaluated by agar cup plate method. Nutrient agar media, Nutrient broth, Sabouraud Dextrose broth (SDB), Sabouraud Dextrose agar (SDA) was used for antibacterial and antifungal activity respectively. Streptomycin (100μg/ml) and Fluconazole (25μg/ml) in DMSO used as reference standards. The plates containing bacterial organism were incubated at 37°C for 24 hours and plates containing fungal organism were incubated at 27°C for 48 hours. The zone of inhibition was calculated by measuring the diameter of the zone of bacterial and fungal growth around the disc. An average of three independent determinations was recorded.

Determination of Minimum Inhibitory Concentration (MIC):

The MIC was determined by the micro dilution method using liquid media with different aliquots of the test material. 5ml of sterilized double strength nutrient broth was poured into sterilized test tube. Test solution of the extract of the concentration ranging from 100 and 200μg/ml were prepared in DMSO/Water.

0.1ml of the inoculum from standardized culture of test organism was spread uniformly. Wells were prepared by using a sterile borer of diameter 10mm and 100μl of the test substance, standard antibiotic and the solvent control were added in each well separately. The plates were placed at 4°C for 1 hour to allow the diffusion of test solution into the medium and plates were incubated at a temperature optimal for the test organism and for a period of time sufficient growth of at least 10 to 15 generations. The zone of inhibition of microbial growth around the well was measured in mm.

RESULTS AND DISCUSSION:

The inhibitory effect of three extracts of the leaves of Muntingia calabura, Basella alba and Basella rubra against pathogenic species in nutrient agar shown in Table 1.

In MIC, the test drug Muntingia calabura exhibited mild inhibitory properties against S.aureus, B.subtilis and E.coli. The test drug Basella rubra exhibited mild inhibitory properties against S.aureus, but in other organisms it failed to show the activity. The test drug Basella Alba exhibited mild inhibitory properties against fungi Aspergillus niger, but it failed to show the activity in both gram +ve and gram –ve organisms. Finally all concentrations Muntingia calabura, Basella alba and Basella rubra possessed noticeable antimicrobial activity against Gram positive, Gram negative bacteria and Fungi, when compared with standards.

Above results suggested that significant antimicrobial activity may be due to the presence of phytoconstituents like phytosterol, triterpenoids, saponins, and tannins in methanolic extracts.

REFERENCES:

1. Pharmacognosy kokate C.K, Purohit A.P and Gokhale S.B.10^th edition.

2. Huddleson F., Dufrain J., Barooms K., and Giefel M., Antibacterial substances in plants J Am Vet. Assoc. 1944, 105, 394-397.

3. Carlson H.J., Bissell H.D. and Meller M. Antibacterial substances separated from Plants J.bact.1946, 52, 155-168.

4. J.F. Morton, Fruits of warm climates, 1987, 65.

5. Zainul Amiruddin Zakaria, Med. Princ. Pract, 2007, 16, 130-136.

6. Ayesha Siddiqua, Premakumari K.B., Roukiya sultana, Vithya and Savitha. Antioxidant activity and estimation of total phenolic content of Muntingia calabura Jan-March 2010 Vol.2, N0.1, pp 205-208.

7. ICRAF, Nairobi, Kenya. Promoting the conservation and use of underutilized and neglected crops 16, IPGRI, Rome, Italy August 1995, Pp.171

8. Chemical and pharmaceutical Bulletin 49 (6) 770-776.

9. Ali MS, Ahmad VU, Usmanhani K, Azhar I and Amtul Z. Antimicrobial screening of some Caesalpiniaceae, Fitoterapia. 1999, 70; 299-304.

Received on 24.02.2010

Accepted on 28.03.2010

© A&V Publication all right reserved

Research Journal of Pharmacognosy and Phytochemistry. 2(3): May-June 2010, 246-248