Antimicrobial activity of leaves extracts of Passiflora foetida

 

Swapnali Mohite*, Rutuja Shah, Naziya Patel

Adarsh College of Pharmacy, Vita, Tal. Khanapur, Dist. Sangli, Maharashtra, India.

 

ABSTRACT:

The present study was carried out to screen and evaluate antimicrobial activity of leaf extract of Passiflora foetida. Aqueous extract of leaves of Passiflora foetida was tested against Escherichia coli, Staphylococcus aureus, Bacillus spp., Pseudomonas aeruginosa which are known to be resistant to various antibiotics. A Passiflora foetida material which was used during this study collected from Kolhapur district. In this study, we examined the in-vitro effect of extracts of Passiflora foetida on Escherichia coli, Staphylococcus aureus, Bacillus spp., Pseudomonas aeruginosa. Aqueous extracts was prepared from fresh Passiflora foetida leaves. The susceptibility of tested bacteria to extract was determined by measuring the diameter of inhibition zones formed around well in plate. The results showed that the Passiflora foetida leaves possess antimicrobial activity.

 

 

 

INTRODUCTION:

Human infections particularly those involving micro- organisms i.e. bacteria, fungi, viruses, nematodes, they cause serious infections in tropical and subtropical countries of the world. In recent years, multiple drug resistance in human pathogenic microorganisms has been developed due to indiscriminate use of commercial antimicrobial drugs commonly used in the treatment of such diseases. Over the last three centuries, intensive efforts have been made to discover clinically useful antimicrobial drugs^{1, 2}. Plants are known to produce phytochemicals, which are potential sources of anticarcinogenic, anticancer, antimicrobial, and antioxidant activity.

 

The antimicrobial activities of medicinal plants extracts have been linked to the presence of bioactive compounds such as tannins, flavonoids and alkaloids which sometimes serve to protect the plants themselves against bacteria, fungi and viral infections as well as exhibiting their antimicrobial properties on these organisms³. The passion fruit tree (Passiflora) belongs to family Passifloraceae⁴. There are two recognized forms of passion fruit: purple (P. edulis Sims) and yellow (P. edulis var. flavicarpa)⁵.

 

In this study, we concentrate on the antibacterial activity of Passion flower (Passiflora species), which is an exotic and fast-growing perennial, vine, occurring in west USA and extend to the Asian countries like India. One of the most important and common species -Passiflora foetida was chosen in this study. The ethno botanical views of P. foetida, reports the decoction of leaves and fruits to treat asthma and biliousness, leaves and root decoction is emmenagogue, used in hysteria ⁶ and leaf paste is applied on the head for giddiness and headache ⁷. The major phyto- constituents of this plant contain alkaloids, phenols, glycoside flavonoids and cyanogenic compounds ⁸.

 

Information of Bacteria:

1. Escherichia coli- :

Escherichia coli is a gram negative bacteria facultatively anaerobic, rod shaped coli form bacteria of the genus Escherichia that is commonly found in the lower intestine of worm blooded organism. Most the E.coli strains are harmless but some serotypes can cause serious food poisoning in their host and are occasionally responsible for product recall due to food contamination the harmless straits are the part of normal flora of gut and can benefit their host by producing vitamin K. E.coli and other facultative anaerobes constitute about 0.1% of gut flora and fecal – oral transmission is the major route through which pathogenic strains of bacterium cause disease. Cells are able to survive outside the body for a limited of time which makes them potential indicator organism to test the environmental sample for fecal contamination.

 

2. Pseudomonas aeruginosa-:

Pseudomonas is a genus of gram negative, aerobic gamma proto bacteria belong to family Pseudomonadaceae and containing 191 validly described species, the member of the genus demonstrates a great deal of metabolic diversity and consequently are able to colonize a wide range of niches there ease of culture in-vitro and availability of an increasing number of Pseudomonas strains genome sequence has made a genus an excellent focus for scientific research the best studied species include P. aeruginosa in its role as an opportunistic human pathogen, the plant pathogen P.syringae, the soil bacterium P.putida and the plant growth promoting P.fluorescenes. Bacteria of their wide spread occurrence in water and plant seed such as dicots, the pseudomonas were absorbed early in the history of microbiology.

 

The generic name Pseudomonas created  for this organisms was defined in rather vague terms by walter migula in 1894 and 1900 as a gram negative rod shaped and polar flagellated bacteria with some sporulating species, the statement was later provided incorrect and was due to refractive granules of reserve material.

 

3. Staphylococcus aurous:

Staphylococcus is a genus of gram positive bacteria under microscope; they appear round and form in grape like clusters. The staphylococcus genus includes at least 40 species of this, nine have to subspecies, one has three subspecies and one has four subspecies. Most are harmless and reside normally on the skin and mucous membrane of human and other organisms found worldwide they are a small component soil microbial flora.

4. Bacillus spp.:

Bacillus is a rod shaped bacterium. Bacillus is found in many different taxonomic groups of bacteria. The Bacillus capitalized and italicized refers to specific genus of bacteria^{9, 10}.

 

MATERIALS AND METHODS:

The plant sample of Passiflora foetida, the leaf was used in this research work was collected from Kolhapur region, and authenticated from Department of Botany, Balwant College of Science, Vita. All other chemicals used were of analytical grade.

 

Equipments:

The equipment used was an analytical balance, bottles, funnel, rotary vacuum evaporator, petri dish, glass beakers, centrifuge, incubators, electric cooker, autoclave, hot air oven.

 

Experimental Methods:

1.    Cultivation of microorganism on suitable media:

a)    Vogel Johanson Medium- This medium contains more tellurite than Baired-Parker medium and is highly effective for Staphylococci with few other microorganisms able to grow within a 24 hrs incubation period.

b)   MacConkeys agar- This is used for the isolation of the E-coli because different types of formulation of this medium are available with the different degree of selectivity. The basis of selectivity is the inclusion bile salt which inhibits non intestinal bacteria and crystal violet inhibits cocci. Lactose is added in MacConkeys medium acid product of fermentation detected by using neutral rod is pink in acid and orange alkaline condition. So E-coli other lactose fermenter produces rod colonies and impart red colour to the surrounding medium after 48 hrs.

c)    Nutrient agar- This is used for isolation of Bacillus bacteria because bacillus bacteria it is simple medium used for the growth purpose bacillus bacteria ^{11, 12}.

 

2.    Extraction of plant materials:

20 g amount of the powdered leaves was weighed and percolated in 200 ml of 96% ethanol contained in 500 ml conical flask. The flask was agitated manually several times over a period of 24 h. The extract was filtered using Whatman No. 1 filter paper and the filtrate collected in a clean beaker was concentrated to dryness by evaporation over a steam bath at 80°C. The aqueous extract was similarly prepared using 20 g of the powdered leaves material in 200 ml of distilled water.

 

 

 

3. Antimicrobial activity of specific medicinal plant on organisms:

A. Preparation of agar plate:

i.     MacConkeys agar = 100 ml distilled water + 5.53 gm HIMEDIA of MacConkeys agar medium

ii.    Cetrimide agar = 100 ml distilled water + 4.67 gm HIMEDIA of Cetrimide agar medium

iii.  Vogel Johanson agar   medium = 100 ml distilled water + 6.1 HIMEDIA Vogel Johanson of agar medium

iv.  Nutrient agar = 100 ml distilled water + 2.8gm HIMEDIA of Nutrient agar medium

Take 4 conical flasks for preparation of media. All media were prepared as mentioned above formula, then mix thoroughly. All prepared media were sterilized in autoclave at 121^◦C/15 LBS pressure for 15 min. After that pour sterilized media in sterilized plates slowly before they solidify.

 

A)  Spreading and preparation of well:

Saline water was taken in that particular bacteria were inoculated and then suspension was spread on agar plates with the help of sterile glass spreader. Then prepare a well with the help of cork borer. Then plant extract was added to the well in aseptic condition.

 

B)   Incubation:

Incubation of plates was carried out at 37⁰C for 48 hrs. After 48 hour remove the plate from incubator and the inhibition zone were observed. Then zones were measured in mm ^{13, 14}.

 

RESULTS:-

Table 1: Results of inhibition zone in mm

Sr.No.	Name of Bacteria	Inhibition zone in mm
1)	Escherichia coli	12
2)	Bacillus spp.	10
3)	Staphylococcus aureus	13
4)	Pseudomonas aeruginosa	08

 

 

Fig. no. 1 Staphylococcus aureus on Vogel Johanson agar

 

Fig. no. 2 Bacillus spp. on Nutrient agar

 

 

Fig. no. 3 Escherichia coli on MacConkeys agar

 

 

Fig. no. 4 Pseudomonas aeruginosa on Cetrimide agar

 

DISCUSSION:

From above study it was concluded that aqueous extract of leaves of Passiflora foetida possess good antimicrobial activity and Pseudomonas aeruginosa is more sensitive towards Passiflora foetida.

 

 

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Accepted on 12.02.2018            © A&V Publications All right reserved