Ethnobotanical Leaflets 13: 472-74 , 2009.

 

 

 

Invitro Antimicrobial Activity and Phytochemical Analysis of Ficus religiosa L. and Ficus bengalensis L. against Diarrhoeal Enterotoxigenic E. coli

 

Uma, B.*,  Prabhakar, K. and Rajendran, S.

 

Division of Microbiology, Rajah Muthiah Medical College and Hospital,

Annamalai Nagar Tamil Nadu, India, 608002

* Corresponding Author: E mail : amu_sri2003@yahoo.co.in

 

Issued 01 April 2009

 

Abstract

The barks of Ficus religiosa L. and Ficus bengalensis L., which belongs to family Moraceae, were investigated for invitro antibacterial activity and phytochemical analysis. The various solvents extract like aqueous, methanol, chloroform, petroleum ether and hexane were screened for antibacterial activity against Enterotoxigenic E. coli isolated from diarrhoeal patients. The preliminary phytochemical analysis of the methanol extracts of both the plants showed the presence of carbohydrates, flavonoids, aminoacids, steroids, saponins and tannins. The extracts were subjected for antibacterial activity against Enterotoxignic E.coli (ETEC) at 200mg/ml concentration by disc diffusion method. The results of antibacterial activity revealed that methanol extracts of both the plants barks exhibits good activity compared to chloroform and aqueous extracts. Petroleum ether and hexane extracts did not show any activity. The antibacterial activities of extracts were compared with standard antibiotics.

Key words: Ficus religiosa, Ficus bengalensis, diarrhoea, Disc diffusion Assay, medicinal plants.

Introduction

      Diarrhoea is one of the most common causes of morbidity and mortality among infants and children in developing countries. Although commensal representatives found in the intestinal flora of humans are non-pathogenic, certain strains are highly pathogenic. Enterotoxigenic E. coli (ETEC) is the most prevalent among the various types of diarrhoeagenic E. coli in India (Taneja et al., 2004).  The increasing antibiotic resistance to commonly used antibiotics exhibited by diarrhoeal pathogens has led to the screening of several medicinal plans for their potential antimicrobial activity (Mukherjee, et al., 1998). Plants are rich in a wide variety of secondary metabolites, such as tannins, terpenoids, alkaloids, and flavonoids, which have been found invitro to have antimicrobial properties (Majorie Murphy Cowan.1999). So, the use of and search for drugs and dietary supplement derived from plants have accelerated in recent years.

       Ficus religiosa. L. belongs to the family Moraceae, is commonly known as Peepal tree, and has many medicinal properties. The barks have been used for diarrhoea, dysentery, leucorrhea, menorrhagia, for vaginal and other urogential disorders. Ficus bengalensis belongs to the family Moraceae, which is commonly known as Banyan tree. It is used in Ayurveda for treatment of diarrhoea, piles, teeth and skin disorders (Warrier et al., 1995). The present study was aimed to carry out the preliminary phytochemical analysis and to screen invitro antibacterial activity against diarrhoeal Enterotoxigenic E. coli (ETEC) isolated from patients.

Materials and Methods

      The barks of F. religiosa and F. bengalensis were collected in and around Chidambaram, Tamilnadu, India and identified, confirmed and authenticated by the Department of Botany, Annamalai University, Tamilnadu, India. The barks were washed, shade dried and extracted with aqueous, methanol, chloroform, petroleum ether and hexane for 48 hours with occasional shaking in a beaker. The extracts were filtered. The filtrate was dried at 50 to 60 o. The extracts were dried and percentage yield was calculated and subjected to preliminary phytochemical analysis. The invitro screening of antibacterial activity was carried out using three Enteroxigenic E.coli (ETEC), isolated from diarrhoeal patients, attending Rajah Muthiah Medical College and Hospital, Annamali Nagar, Tamilnadu India.

       The antibacterial screening of the extracts were carried out by determining the zone of inhibition using disc diffusion method (Sahoo et al., 2006). The strains were grown to logarithmic phase in nutrient broth and the inoculum was prepared by adjusting the turbidity of bacterial suspension to 0.5 McFarland’s tube with nutrient broth (Mc Farland et al., 1987).

       The dried extracts were dissolved in 5% Dimethyl sulphoxide (DMSO) to the concentration 200mg/ml and finally sterilized by filtration. The sterile discs (6mm in diameter) were impregnated with 20 l of the above extracts to achieve desired concentration of 4mg/ml. The extract discs were placed on Muller-Hinton agar plates (Himedia), which were previously inoculated with test strains and incubated at 37oC for 24 hours. Amikacin disc (10g) and 5% DMSO impregnated discs were used as positive and negative controls respectively and the zones of inhibition were recorded.

Results

        Preliminary phytochemical analysis of the methanol extracts of the barks of F. religiosa and F. bengalensis showed the presence of carbohydrates, flavonoids, aminoacids, steroids, sopaninis and tannins. The antibacterial activity in terms of zone of inhibition is shown in Table 1.

Table. 1   Zone of inhibition of Ficus religiosa. L and Ficus bengalensis L.

Solvent extracts

Conc. of disc

Ficus religosa

Ficus bengalensis

ETEC 1

ETEC 2

ETEC3

ETEC 1

ETEC 2

ETEC3

Aqueous

4mg/ml

8mm

8mm

10mm

8mm

12mm

10mm

Methanol

4mg/ml

12mm

12mm

14mm

16mm

14mm

14mm

Chloroform

4mg/ml

10mm

10mm

12mm

12mm

12mm

12mm

Petroleum ether

4mg/ml

-

-

-

-

-

-

Hexane

4mg/ml

-

-

-

-

-

-

Amikacin

10 g

24mm

22mm

20mm

20mm

24mm

24mm

DMSO

5%

-

-

-

-

-

-

 

Discussion and Conclusion

The use of plants and plant preparations has been in existent since prehistory. The World Health Organization (WHO) reported that about 80% of the world’s population depend mainly on traditional medicine and the traditional treatment involve mainly the use of plant extracts (WHO, 1993). In the present study, among the various solvent extracts, methanol extract found to be more active against all the Enterotoxigenic E. coli, which is isolated from diarrhoeal patients. These findings suggest new pathway in elucidating a potent antimicrobial agent from Ficus religiosa L. and Ficus bengalensis L. in general in Ficus species. Invivo experiments are needed to confirm these findings.

References

1.      Husain,A., Virmani, O.P., Popli, S.P., et al, (1992). Dictionary of Indian Medicinal Plants, CIMAP, Lucknow, India.

2.      Morjorie Murphy Cowan, (1999). Plant Products as antimicrobial agents. Clinical Microbiology Review 12:564-582.

3.      Mc Farland, J., (1987). Standardization of bacterial culture for disc diffusion assay. Journal of America Medical Association 49: 1176-1178.

4.      Mukherjee, P.K., Saha, K., Murugesan, T., Mandal, S.C., Pal, M., Saha, B.P., (1998). Screening of antidiarrhoeal profile of some plant extracts of specific region of West Bengal, India. Journal of Ethno pharmacology 60: 85-89.

5.      Sahoo, S., Kar, D.M., Mohapatra, S., Rout, S.P., Dash, S.K., (2006). Antibacterial activity of Hybanthus enneaspermus against selected UTI pathogens. Indian Journal of Pharmaceutical Sciences 68:653-655.

6.      Warrier, D.K., Nambiar, V.P.K., Ramankutty, C., (1995). Indian Medicinal Plants   (1-5) Orient Longman Ltd, Madras.

7.      WHO, 1993. Summar 9 WHO guidelines for the assessment of herbal medicines. Herbal Grom, 28, 13-14.

8.      Taneja, N., Mohan, B., Khurana, S., Sharma, M., (2004). Antimicrobial resistance in selected bacterial enteropathogens in North India, Indian Journal of Medical Research 120: 39-43.