Antibacterial Activity of Cardiospermum halicacabum and Melothria heterophylla
1Mariyappan M., 1Bharathidasan R., 1Mahalingam R., 2Madhanraj P., 1Panneerselvam A. and
*1Ambikapathy V.
1P.G.Research Dept.
of Botany and Microbiology, A.V.V.M. Sri Pushpam
College (Autonomous), Poondi – 613503, Thanjavur. DT, Tamil Nadu, India.
2Dept of
Microbiology, Thanthai Hans Roever
College of Arts and Science, Perambalur - 621 212,
Tamil Nadu, India.
*Corresponding
Author E-mail: drva1967@yahoo.com
ABSTRACT:
The bacterial organisms were isolated from
drinking water (Bacillus, Borchothrix, Clavibacter sp,
Ancylobacter sp, and Brevi
bacterium). Selected Indian medicinal plants Cardiospermum
halicacabum and Melothria
heterophylla were selected for antibacterial
studies. The solvents used for the extraction of plant roots were n- butanol, ethyl acetate and distilled water. The invitro antibacterial activity was performed by agar well
diffusion method. The most susceptible Gram-Postive
bacteria was Bacillus sp, Brevibacterium sp, and the most
susceptible Gram-negative bacteria was Borchothrix
sp, Clavibacter
sp, and Ancylobacter
sp. The extracts of plant Cardiospermum halicacabum
and Melothria heterophylla
inhibited the growth of the bacterial strains investigated. The most active
extracts was compared with the standard antibiotics, penicillin, Streptomycin
and Ampicillin 100mg/disc). The results obtained in the
present study suggest that Cardiospermum halicacabum and Melothria
heterophylla could be used in treating diseases
caused by the test organisms. The results are discussed in detail.
KEYWORDS:
Medicinal Plants,
antibacterial activity, aqueous extract, n-butanol
excretory ethyalcetate extract
INTRODUCTION:
Medicinal plants are a source of great
economic value in the Indian subcontinent. Nature has bestowed on us a very
rich botanical wealth and a large number
of diverse types of plants grow in different parts of the country. India is
rich in all the three levels of biodiversity, namely species diversity, genetic
diversity and habitat diversity. In India thousands of species are known to
have medicinal value and the use of different parts of several medicinal plants
to cure specific aliments has been in vogue since ancient times. Herbal
medicine is still the mainstay of about 75-80% of the whole population, mainly
in developing countries. For primary health care because of better cultural
acceptability better compatibility with the human body and fewer side effects.
However, the last few years have seen a
major increase in their use in the
developed world1.
Infectious diseases are the leading cause of
death world –wide. Antibiotic resistance has become a global concern2.
The Clinical efficacy of many existing antibiotics is being threatened by the
emergence of multidrug – resistant pathogens3. Many infection
diseases have been known to be treated with herbal remedies throughout the
history of mankind. Natural products, either as pure compounds or as
standardized plant extract, provide unlimited opportunities for new drug leads
because of the unmatched availability of chemical diversity. There is a
continuous and urgent need to discover new antimicrobial compounds with diverse
chemical structures and novel mechanisms of action for new and re–emerging infectious diseases4 Therefore
researchers are increasingly turning their attention to folk medicine, looking
for new leads to develop better drugs against microbial infection5.
The increasing failure of chemotherapeutics and antibiotic resistance exhibited
by pathogenic microbial infectious agents has led to the screening of several
medicinal plants for their potential
antimicrobial activity6,7. India is a varietal emporium of
medicinal plants and is one of the richest countries in the world with regard
to genetic resources of medicinal plants. It exhibits a wide range in
topography and climate, which has a bearing on its vegetation and floristic
composition moreover, the agro-climatic conditions are conducive for
introducing and domesticating new exotic plant varieties8
PLANT MATERIALS COLLECTION:
The plant species namely Cardiospermum,
halicacabum and Melothria
heterophylla were collected in Thanjavur –karupamudaliyar kottai. The collected samples were carefully stored in
sterile polythene bags and used for the present study.
Sterilization of plant materials:
About 1gram of fresh and healthy roots was
taken for each solvent including aqueous. The roots of both plants (Cardiospermum hailcacabum
and Melothria heterophylla
) were sterilized with running tap water and soaked in 0.1% mercuric chloride
Finally, the roots were washed with distilled water ( three times ) and
shade dried.
Composition of nutrient Agar medium:
Chemicals Composition
Beef extract - 3g
Peptone - 5g
Sodium chloride - 5g
Agar - 20g
Distilled water - 1000ml
pH - 7.0
Preparation of plant root extract:
About one gram of sterilized roots were
ground in mortar and pestle with 10ml of aqueous and organic solvents (ethyl
acetate, and n-butanol it was filtered through Whatman No1 filter paper, the supernatant was
collected and stored for antibacterial screening
Antibacterial activity (Agar –well diffusion
method ):
The antibacterial activities of the roots
were tested against the selected bacterial strains. The 20ml of sterilized agar
medium was poured into each sterile petriplates and.,
allowed to solidify. The test bacterial cultures were evenly spread over the
appropriate media by using sterile cotton, Swab. Then a well of 0.5 cm was made
in the medium by using a sterile cork borer, 150 µl of each ethylacetate,
n-butanal and aqueous plant extracts were transferred
into separated wells. After these plates was incubated at 37oC for
24-48 hours. After incubation period, the results were observed and measured
the diameter of inhibition zone around the each well.
RESULT AND
DISCUSSION:
In the present investigation, the anti
bacterial properties of ethyl acetate, n – butanol
and aqueous extracts of two medicinal plants Cardiospermum
halicacabum and Melothria
heterophylla viz.., were tested against five
human pathogenic bacteria. The antibacterial properties of the two extracts of Cardiospermum
halicacabum were also comparatively analysed sensitivity test.
Antibacterial activity of Cardiospermum halicacabum:
The ethylacetate
extract of Cardiospermum halicacabum, exhibited maximum zone of inhibition
against Bacillus (17mm), Brochothrix (18mm), Clavibacter (16mm), Ancylobacter
(20mm) and Brevibacterium (20mm) were observed.
The n- butanol
extract of Cardiospermum halicacabum, showed maximum zone of inhibition against Bacillus
(25mm), Borchothrix (30mm), Clavibacter (30mm), Ancylobacter
(25mm) and Brevibacterium (30mm). The aqueous extract of root showed the
inhibition zone diameter ranging from 13-16mm against tested bacterial
pathogens (Table-2).
Antibacterial activity of Melothria heterophylla:
The ethylacetate
extract of Melothria heterophylla
exhibited maximum zone of inhibition against Bacillus (12mm), Ancylobacter (10mm) was observed.
The n-butanol
extract of Melothria heterophylla
showed maximum zone of inhibition against Bacillus (18mm), Borchothrix
(25mm), Clavibacter (25mm), Anchylobacter
(30mm) and Brevibacterium (30mm).
The aqueous extract of root showed no zone
inhibition against tested bacterial pathogens (Table-2).
Antibiotic sensitivity test (positive
control ):
The antibiotic sensitivity test using
standard antibiotics, viz.., streptomycin, ampicillin
and penicillin were tested against both bacteria studied. The results of
antibiotic sensitivity represented (Table-3).
All the antibiotics used were exihibited higher antibacterial activity. The results
confirmed that both the solvent extract of Cardiospermum
halicacabum and Melothria
heterophylla exihibited
a higher antibacterial activity against Bacillus,Borchothrix,
Clavibacter, Ancylobacter
and Brevibacterium.
Similarly. when compared to the standard
antibiotics. the solvent extract of Cardiospermum
halicacabum and Melothria
heterophylla showed higher antibacterial activity
against the bacteria (Table-3).
Antibacterial effect of solvents (negative
control):
The antibacterial effect of ethyl acetate,
n-butanol, and water solvents revealed no activity
against bacteria studied.
The maximum antibacterial activity was shown
by Cardiospermum halicacabum
and Melothria heterophylla,
reapectively. The methanol extracts of the
investigated plants showed maximum antibacterial activity against gram-negative
Ancylobacter, similar results were also
reported by venkatesan et al 20069.
In the present study Cardiospermum helicacabum plants showed maximum inhibition of 30mm
and minimum of 16mm.
Table-1Antibacterial activity of Cardiospermum halicacabum
plant extract.
|
S. No |
Name of the organisms |
Organic Solvents |
||
|
Ethyl acetate |
n-butanol |
Distilled water |
||
|
Zone of inhibition (mm ) |
||||
|
1 2 3 4 5 |
Bacillus subtilis Borchothrix campestris Clavibacter iranicus Ancylobacter aquaticus Brevibacterium linens |
17mm 18mm 16mm 20mm 20mm |
25mm 30mm 30mm 25mm 30mm |
13mm 15mm 12mm 17mm 16mm |
Table- 2 Antibacterial activity of Melothria heterophylla
plants extract.
|
S. No |
Name of the organisms |
Organic Solvents |
||
|
Ethyl acetate |
n-butanol |
Distilled water |
||
|
Zone of inhibition (mm ) |
||||
|
1 2 3 4 5 |
Bacillus subtilis Borchothrix campestris Clavibacter iranicus Ancylobacter aquaticus Brevibacterium linens |
12mm - - 10mm - |
18mm 25mm 25mm 30mm 30mm |
- - - - - |
Antibiotic sensitivity test on bacteria
(positive control)
|
S. No |
Name of the organisms |
Streptomycin |
Ampicillin |
Penicillin |
|
Zone of inhibition (mm ) |
||||
|
1 2. 3 4 5 |
Bacillus subtilis Borchothrix campestris Clavibacter iranicus Ancylobacter aquaticus Brevibacterium linens |
13 11 12 10 - |
10 10 - 8 - |
10 10 - 12 - |
The potential for developing antibacterial
from higher plants appears rewarding as it will lead to the development of phylomedicine to act against microbes. Plant-based
antibacterial have enormous therapeutic potential as they can serve the purpose
with lesser side effects that are often associated with synthetic antibacterial
(Lwu et al 1999)7.
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Received on 11.10.2011 Accepted on 15.11.2011
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Press All Right Reserved
Asian J. Pharm.
Res. 1(4): Oct. - Dec. 2011;
Page 111-113