In vitro Screening of Antifungal Activity of Methanol Extract of Plumbago indica L. against some pathogenic species of fungi

 

Dibyajyoti Saha*, Swati Paul

Department of Pharmacy, BGC Trust University Bangladesh Chittagong

*Corresponding Author E-mail: saha.dibyajyoti@gmail.com

 

ABSTRACT:

The main objective of the present study was to determine the methanolic extract of Plumbago indica L. for antifungal activity. To determine the antifungal activity, agar disc diffusion method was used. The antifungal activity of the extracts was compared with standard drug Fluconazole (500 μg/disc). The methanol extract of Plumbago indica L. showed very good antifungal activity ranging from zone of inhibition (10.0-27.0) mm and Candida albicans was the most susceptible fungal strain of the methanolic extract of. Plumbago indica L .Due to these promising results, further in vivo studies over Plumbago indica L. must be conducted.

 

KEYWORDS Plumbago indica L.; methanol extract; antifungal activity.

 


INTRODUCTION:

Vast natural resources of medicinal plants are being used for thousands of years for the cure of many diseases in all over the world. If we could use medicinal plants properly we could get medicines at low cost and then it might be possible to fulfill the demand of our medication. This will supply low cost medicine to our poor people and we could establish a better health care system [1]. Recently, some higher plant products have attracted the attention of microbiologists to search for some phytochemicals for their exploitation as anti-microbials. Such plant products would be biodegradable and safe to human health [2] . Plumbago indica (Begali name: Agnichita ) belonging to the family Plumbaginaceae , is a family of flowering plants, with a cosmopolitan distribution. The family is sometimes referred to as the leadwort family or the plumbago family. Most species in this family are perennial herbaceous plants, but a few grow as lianas or shrubs. The plants have perfect flowers and are pollinated by insects. They are found in many different climatic regions, from arctic to tropical conditions, but are particularly associated with salt-rich steppes, marshes, and sea coasts. Plumbago popularly known as chittiramulam, in Tamil and white leadwort in English.

Plumbaginaceae is distributed as a weed throughout the tropical and subtropical countries of the world. The familyPlumbaginaceae consists of 10 genera and 280 species. The genus Plumbago includes 3 species, namelyPlumbago indica. L, Plumbago rosea. L, Plumbagocapensis. L, and Plumbago zeylanica .L, which are distributed in several parts of India [3]. Plumbago Indica root increases digestive power, promotes appetite and has long been marked as a powerful antiseptic. A liniment made from bruised root mixed with a few amount of bland oil is used in treating rheumatism, paralysis, leucoderma, enlarged glands and buboes and scorpion-sting [4]. Scraped root is inserted into the mouth of the womb to procure illegal abortion, a tincture of the root is used in secondary syphilis, leprosy, dyspepsia, hemorrhage, piles, flatulence, loss of appetite and other digestive complaints, and the milky juice of the plant is used in ophthalmia, scabies and as an antiseptic agent. In order to establish the above assertion about their validity, these medicinal plants must be subjected to extensive study in different research works. For this reason, Alpinia conchigera Griff. and Plumbago indica L. (Plumbaginaceae) two Bangladeshi plants under the family of Zingiberaceae and Plumbaginaceae, respectively were selected and subjected for chemical, biological and pharmacological investigations to explore the antimicrobial, cytotoxicity, antidiarrhoeal, anti-motility, analgesic activity of methanol extracts of the above mentioned plant species.

 

MATERIALS AND METHODS:

Collection of Plant material

The plants selected for present work Plumbago indica. L (Family: Plumbaginaceae ) and was collected from Naramuk, Rajsthali of Rangamati district. After collection, suitable herbarium sheet for each plant with some general information were prepared and send to Bangladesh Council

of Scientific and Industrial Research (BCSIR), Baluchara, Chittagong for identification. They provided us the scientific name of the plants.

 

Extraction

The collected plant (leaves and stems) was separated from undesirable materials or plants or plant parts and was shed-dried (35-50c). The plant was ground into a coarse powder with the help of a suitable grinder. The powder was stored in an airtight container and kept in a cool, dark and dry place until extraction commenced. About 185 gm of powdered plant material of Plumbago indica L. (Family: Plumbaginaceae ) was was taken in a clean, flat bottomed amber glass container and soaked in 1700ml of methanol The container with its contents was sealed and kept for a period of 10 days accompanied by continuous shaking. The whole mixture then underwent a coarse filtration by a piece of clean, white cotton materials. Then they were filtered by using Whatman filter paper number 1 and the solvent was made to evaporate under the room temperature. The obtained extract was collected .The residues were stored in a refrigerator until further studies.

 

Fungal Strains

The antifungal activity of plant extract were investigated against six pathogenic fungal strains such as Aspergillus niger, Blastomyces dermattitidis, Candida albicans, Pityrosporum ovale, Trichophyton spp, Microsporum spp., Cryphcoccus neoformans. All the fungal strains were collected from Bangladesh Council of Scientific and Industrial Research (BCSIR), Bangladesh.

 

Antifungal Assay

In vitro antifungal screening was performed by disc diffusion assay method [5, 6] where Potato Dextrose Agar (PDA) medium was used for the antifungal activity.

 

Their antifungal activity were tested against six fungal strains at a concentrations of 250 μg/disc, 500 μg/disc for each and the results were compared with griseofulvin (500 μg/disc). The activity was determined after 72 hours of incubation at 37.5oC.

 

Preparation of the medium

The weight amount of potato slice was boiled with a little amount of distilled water for 30 minutes and applied for course filtration by the help of cotton. The required amount of dextrose and bacterial agar medium were properly mixed in a conical flask. Finally the constituents of two flask were mixed thoroughly after the adjustment of volume by the distilled water the medium was sterilized in an autoclave. The pH of the medium was adjusted to 5.6.

 

Table- 1: Composition of the Potato Dextrose Agar (PDA) medium

Potato Dextrose Agar (PDA) medium (1000 ml)

Ingredients

Amount

Potato slice

200.0 gm

Dextrose

20.2 gm

Bacterial agar medium

16.0 gm

Distilled water

q.s

 

Result of the antifungal screening

The result of the antifungal screening assay of methanol extract of Alpinia conchigera Griff. against the tested fungal strains were shown in Table- 2

 

Table- 2: Anti-fungal activity of the crude extract of MEPI, standard and blank

Tested fungi

 

Zone of inhibition (mm)

MEPI

S

C

A

B

500 μg/disc

 

Aspergillus niger

14

21

27

-

Blastomycesdermatitides

13.5

25

26

-

Candida albicans

19

27

29

-

Pityrosporum ovale

12

20

22.5

-

Trichophyton spp

13

22

29

-

Microsporum spp

10

25

27

-

Cryphcoccus neoformans

11

20

21

-

 

 

 

 

 

 

 

 

 

 

 

[MEPI = Methanol extract of Plumbago indica L. A = 250μg/disc, B = 500μg/disc, S = Standard (Fluconazole) and C = Control]

 



Figure- 1: Comparison of Zone of Inhibition of different fungi with MEPI and standard


 

DISCUSSION:

The antifungal activities of the crude extracts were evaluated by the disc diffusion method against seven fungal strains using Fluconazole as standards. In the screening, the methanol extract of showed strong antifungal activity with zone of inhibition of (10.0- 27.0) mm respectively while the highest antifungal activity was seen against Candida albicans, Blastomyces dermatitides, Microsporum spp and Trichophyton spp . Candida albicansis the most susceptible fungal strain of the methanolic extract of Plumbago indica L.

CONCLUSION

The result shows that the methanolic extract of Plumbago indica L. possessed antifungal activity against all the tested fungal strains.. So the active principles which are responsible for this antifungal activity is to be explored. The isolation of these active constituents showing antifungal activity can be more useful and work is to be done in this regard.

 

REFERENCES

1. Ghani A. (2003) Medicinal plants of Bangladesh, 2nd Ed. pp.238. The Asiatic Society of Bangladesh, Dhaka.

2. Kumar A, Shukla R, Singh P, Prasad CS, Dubey NK (2008).Assessment of Thymus vulgaris L. essential oil as a safe botanicalpreservative against post harvest fungal infestation of food commodities. Innov. Food Sci. Emerg., 4: 575-580.

3. Van Der Vijver Lm. Distribution of plumbagin in the Plumbaginaceae. Phytochemistry 1974, Vol 11, Pages 32473248.

4. Bala Rathinasabapathi, Walid M. Fouad, and Celia A. Sigua . β-Alanine Betaine Synthesis in the Plumbaginaceae. Purification and Characterization of a Trifunctional, S-Adenosyl-l-Methionine-Dependent N-Methyltransferase from Limonium latifolium Leaves. Plant Physiol. 2001, Vol 126(3), Pages 12411249.

5. Bear AW, kirby WMM, Sherris JC, Turck M. (1966) Antibiotic susceptibility testing by standardized single disc method. Am. J. Clin. Pathol. 44: 493-496.

6. Rios JJ, Reico MC, Villar A. (1988) Antimicrobial screening of natural products. Enthopharmacol. 23: 127-149.

 

 

 

Received on 16.04.2012 Accepted on 22.05.2012

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Asian J. Res. Pharm. Sci. 2(2): April-June 2012; Page 55-57