Antibacterial Activity of Methanolic and Ethanolic Extracts of Leaves and Fruits of Ficus palmata Forssk.

Arun Kumar*, Ruchika Guleria and Anu Bhardwaj

Abhilashi Institute of Life Sciences, Tanda, Ner Chowk, Dist. Mandi, Himachal Pradesh, India

ABSTRACT:

The various parts of Ficus palmata Forssk. has been traditionally used for the treatment of various human diseases. In the present study the antibacterial activity of methanolic and ethanolic extracts of leaves and fruits were assayed against Escherichia coli (Gram negative species) and Staphylococcus aureus (Gram positive species) by disc diffusion method. The extracts (methanolic and ethanolic) of leaves and fruits of this plant exhibit antibacterial activity against Escherichia coli and Staphylococcus aureus indicating their broad spectrum antibacterial properties. Ethanolic extracts of fruits found to have more antibacterial activity against the test organisms then the methanolic extracts. The zone of inhibition observed significantly higher than the standard tetracycline in some ethanolic fruit extracts against both the test bacterial species. The magnitude of antibacterial activity was observed to be more in fruit extracts then the leaves extracts.

KEYWORDS: Antibacterial activity, Latex, Ficus palmata, Zone of inhibition.

The Indian medicine systems such as Ayurveda, Siddha, Unani and homeopathy entirely depends upon the plants and the derivatives obtained from the plants¹. Ficus palmata Forssk.(Fig.1.) is a deciduous, moderate-sized tree, 6 to 10 metres in height; smooth, dull, ash gray, can be stripped off with the hand, exposing the white to light-yellow wood underneath, wood, moderately hard. Leaves are alternate, broad, ovate, membranous,12.92 cm long,14.16 cm broad, having reticulate pinnate venation and dentate margin; dark green and rough on the upper surface, light green and tomentose on the lower surface². Ficus palmata have high medicinal and nutritional value due to the presence of bio-active compounds³.

Fig.1: Plant of Ficus palmata Forssk.

The fruits of Ficus palmata contain chiefly sugars and mucilage and act as a demulcent and laxative. They are principally used as an item of diet in cases of abdominal disorder⁴ and in the diseases of the lungs and the bladder⁵. They are also used as a poultice⁶. Fruits are also recommended for the curing jaundice, oedema, anaemia, leucoderma and vitiligo⁷. Ripe fruits are used in dysentery and vomiting^8,9.

In the past lots of workers have investigated antibacterial activity of various species of Ficus such as in roots of Ficus benghalensis and Ficus racemosa L.¹⁰, leaves and fruits of Ficus auriculata Lour.¹¹, bark extract of Ficus platyphylla¹², leaves extract of Ficus tsiela L.¹³, bark of Ficus glomerata L.¹⁴, methanolic extracts of leaves, fruits and bark of Ficus glomerata L.¹⁵, whole plant of Ficus deltoidea Jack.¹⁶. It is clear that lots of species of Ficus exhibited antibacterial activity, but very little work has been done on the antibacterial activity of Ficus palmata till now to best of our knowledge. Due to various traditional medicinal uses of this plant, the present research work aimed to study the antibacterial activity of various extracts (methanolic and ethanolic) of this plant against Escherichia coli (Gram-negative bacterium) and Staphylococcus aureus (Gram-positive bacterium) causing various human diseases.

MATERIALS AND METHODS:

The collected plant specimen were identified and submitted in the herbarium of Abhilashi Institute of Life Sciences. The fresh leaves and fruits of Ficus palmata Forssk. were collected from village Tanda, Distt. Mandi, Himachal Pradesh (N:31⁰35’27.73’’, E :76⁰53’21.76’’, H: 746m msl) (Fig.2). The freshly collected leaves and fruits were stored at 4°C in aluminum foil.

Village Tanda Distt Mandi

(Himachal Pradesh), INDIA

N : 31⁰35’27.73”

E :76⁰53’21.76”

H: 746m

Fig.2: Place of collection of material.

Preparation of Plant Extract

Collected leaves and fruits from Ficus palmata were dried at room temperature and then ground into fine powder by using the mortal pastel. Then the different amount of powdered leaves and fruits such as 10 mg, 20 mg, 30 mg, 40 mg, 50 mg and 60 mg were mixed with 10 ml of ethanol or methanol for preparing respective extracts. Then mixtures were placed in auto shaker with very low speed for overnight and then filtered through the Whattman’s filter paper and used against the test bacterial species.

Test Organisms Used

In this study two bacterial species Escherichia coli and Staphylococcus aureus were used as test organisms. These were collected from Department of Microbiology, Abhilashi Institute of Life Sciences, Tanda, Distt. Mandi, Himachal Pradesh.

Media Used

In the present study different media like Nutrient broth, Nutrient agar and Eosin Methyl Blue (EMB) were used for culturing of test bacterial species.

Preparation of Suspension of Bacterial Culture

The test organisms used in the present study (Escherichia coli and Staphylococcus aureus) were firstly cultured in nutrient broth, incubated for 24 hrs in incubator shaker at 120 rpm at 37°C ± 2°C.

Determination of antibacterial effect

Paper Disc Method is used to study the antibacterial effect shown by various extracts of leaves and fruits. In this Whattman’s filter paper discs of 6mm in diameter were saturated with above prepared different concentrations of extracts. Sterile nutrient agar (for Staphylococcus aureus) and Eosin Methyl Blue (Escherichia coli) plates were inoculated with test organisms and then discs were placed. These plates were incubated at 37°C ± 1°C for 24 hrs in incubator, after which the zone of inhibition measured.

Statistical analysis

All experiments were carried out in triplicate. Data analysis done by using MS office 2010. Data are presented as arithmetic means and the results obtained were analyzed in terms of standard deviation.

RESULTS AND DISCUSSION:

Leaves of Ficus palmata Forssk.

Methanolic leaf extract of Ficus palmata Forssk. showed maximum inhibition zone 3.3 mm at concentration 60mg/10ml, minimum with inhibition zone 2.3 mm at concentration 20mg/10ml and 50mg/10ml against Escherichia coli respectively. Whereas methanolic leaf extract of 30mg/10ml showed maximum inhibition zone 7.0 mm and minimum inhibition zone 3.3 mm at concentration 10mg/10ml against Staphylococcus aureus.

Table 1: Antibacterial effect of extracts of Leaf of Ficus palmata.

	S. No.	Concentration Leaf: Methanol or Leaf: Ethanol (mg/ 10ml)	Test Bacterial sp.
			*Escherichia coli*	*Staphylococcus aureus*
			Zone of Inhibition(mm) Mean (± SD)	Zone of inhibition (mm) Mean (± SD)
Tetracycline			7.0 (± 0.0)	5.0 (± 0.0)
Methanolic Leaf Extract	1. 2. 3. 4. 5. 6.	10 mg 20 mg 30 mg 40 mg 50 mg 60 mg	NI** 2.3 ( ± 0.4) 2.6 ( ± 0.4) 3.0 (± 0.8) 2.3(± 0.4) 3.3(± 0.4)	3.3 (± 0.4) 4.0 (± 0.8) 7.0 (± 1.4) 0.0( ± 0.0) 6.3( ±2.0) 3.6( ± 0.4)
Ethanolic Leaf Extract	1. 2. 3. 4. 5. 6.	10 mg 20 mg 30 mg 40 mg 50 mg 60 mg	2.3 (± 0.4) 3.0( ± 0.8) 3.6 (± 0.4) 2.6(± 0.4) 2.6(± 0.9) 4.0(± 0.8)	NI** NI NI NI NI 1.3 (± 0.8)

* Data are the arithmetic means ± S.D. n=3

** No Inhibition.

Table 2: Antibacterial effect of extracts of Fruit of Ficus palmata.

No.

Concentration

Fruit: Methanol or Fruit: Ethanol (mg/ 10ml)

Test Bacterial sp.

Escherichia coli

Staphylococcus aureus

Zone of Inhibition(mm) Mean (± SD)

Zone of inhibition(mm) Mean (± SD)

Tetracycline

7.0(± 0.0)

5.0( ± 0.0)

Methanolic Fruit

Extract

10 mg

20 mg

30 mg

40 mg

50 mg

60 mg

2.6 (± 0.4)

3.3 (± 0.9)

3.6( ± 0.4)

4.0(± 0.0)

2.3 (± 0.4)

3.0 ±( 0.5)

4.6 ( ± 0.9)

2.6 (± 0.9)

3.6 ( ± 0.4)

3.6 (± 0.4)

0.0 (± 0.0)

4.3( ± 0.4)

Ethanolic Fruit

Extract

10 mg

20 mg

30 mg

40 mg

50 mg

60 mg

2.3( ± 0.9)

8.0 (±0.8)

6.6( ± 0.4)

7.3( ± 1.2)

8.0( ±1.6)

0.0 (± 0.0 )

6.6 (± 0.4 )

4.6 (± 1.2)

6.6 (± 2.3)

3.3 (± 0.4)

4.6 (± 0.4)

4.6( ± 0.4)

*Data are the arithmetic means ± S.D. n=3

Methanolic extract of 50mg/10ml also effective against Staphylococcus aureus with inhibition zone of 6.3 mm. Whereas no inhibition has been observed in case of Staphylococcus aureus with concentration of 40mg/10ml. Inhibition zone with ethanolic leaf extract of Ficus palmata Forssk. has been observed maximum 4.0 mm in concentration 60mg/10ml and minimum inhibition zone 2.3mm with concentration 10 mg/10ml for Escherichia coli, whereas inhibition zone 1.3 mm has been observed at concentration 60mg/10ml and other concentration of leaf extract has found ineffective against Staphylococcus aureus (Table 1).

Fruits of Ficus palmata Forssk.

Methanolic fruit extract of Ficus palmata Forssk. showed maximum antibacterial activity with inhibition zone 4.0 mm at concentration 40mg: 10ml and minimum antibacterial activity with inhibition zone 2.3 mm at concentration 50mg:10ml against Escherichia coli, inhibition zones of 3.6 mm, 3.3 mm, 3.0 mm and 2.6 mm has been observed with methanolic fruit extract concentrations of 30mg/10ml, 20mg/10ml, 60mg/10ml and 10mg/10ml respectively against Escherichia coli. Whereas methanolic fruit extract of 10mg/10ml showed maximum antibacterial activity with inhibition zone 4.6 mm and minimum 2.6 mm against Staphylococcus aureus, On other hand inhibition zone of 4.3mm, 3.6mm and 3.6mm has been observed with methanolic extract concentrations of 60mg/ml, 30mg/ml and 40mg/ml. Extract concentration 50mg/10ml does not show any effect against Staphylococcus aureus.

Ethanolic fruit extract of Ficus palmata Forssk. showed maximum antibacterial activity with inhibition zone 8.0 mm at concentration 20mg:10ml and 50mg:10ml, whereas minimum inhibition zone 2.3 mm has been observed at concentration 10mg:10ml against Escherichia coli, Other extract concentrations such as 40mg/10ml and 30mg/10ml exhibited inhibition zone of 7.3mm and 6.6mm against Escherichia coli, extract concentration of 60mg/10ml does not inhibit Escherichia coli growth. Ethanolic extract of concentrations of 10mg/10ml and 30mg/ml showed maximum inhibitions of 6.6 mm against Staphylococcus aureus and minimum inhibition zone 3.3mm has been observed with concentration 40mg/10ml. Whereas other concentrations 20mg/10ml, 50mg/10ml and 60mg/10ml exhibited antibacterial activity with inhibition zone of 4.6 mm. (Table 2).

DISCUSSION:

Methanolic as well as ethanolic extracts of leaves and fruits of Ficus palmata exhibited antibacterial activity against both types of bacterial species i.e. Escherichia coli (Gram negative species) and Staphylococcus aureus (Gram positive species) indicating that these parts of plant having broad spectrum of antibacterial activity. Such results also showed in the only single research work reported on this plant in terms of antibacterial activity in 2011¹⁷. Magnitude of antibacterial activity has been observed high in methanolic and ethanolic extracts of fruits of this plant. Whereas leaves extracts also showed antibacterial activity but with low intensity against test organisms. The ethanolic extract of fruits at concentrations 20mg/10ml and 50mg/10ml found to be most effective against Escherichia coli , where as ethanolic extract concentrations 10mg/10ml and 30mg/10ml found to be effective against Staphylococcus aureus. The magnitude of antibacterial activity in these concentrations found to be significantly higher than the tetracycline a well known antibiotic.

CONCLUSION:

Leaves and fruits of Ficus palmata have antibacterial activity against both Gram negative and Gram positive bacteria. Out of these two parts taken fruits possess high antibacterial activity and its ethanolic extracts found to be most effective.

ACKNOWLEDGEMENT:

Authors thanks the Department of Botany, Abhilashi Institute of Life Sciences (affiliated to Himachal Pradesh University, Shimla, Himachal Pradesh) for providing all the support for this work.

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Received on 20.09.2012

Modified on 03.10.2012

Accepted on 25.10.2012

Research Journal of Pharmacognosy and Phytochemistry. 4(6): November – December 2012, 310-313