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I nternational J ournal of Universal Pharmacy and Bio Sciences 3(1): J anuary-February 2014
INTERNATIONAL JOURNAL OF UNIVERSAL
PHARMACY AND BIO SCIENCES
IMPACT FACTOR 1.89***
ICV 5.13***
Bio Sciences RESEARCH ARTICLE!!!


EVALUATION OF ANTIBACTERIAL ACTIVITY OF DIFFERENT PARTS
OF MEDICINAL FERN MARSI LEA MI NUTA L.
G. Nagarajan
1
and A. John De Britto
1
*
Plant Molecular Biology Research Unit,
Post Graduate and Research Department of Plant Biology and Plant Biotechnology,
St.Xavier's College (Autonomous), Palayamkottai - 627 002, Tamil Nadu, India


KEYWORDS:

Antibacterial activity,
medicinal fern, Marsilea
minuta.
For Correspondence:
A. John De Britto *
Address: Post Graduate
and Research Department
of Plant Biology and
Plant Biotechnology,
St.Xavier's College
(Autonomous),
Palayamkottai - 627 002,
Tamil Nadu, India.
*E-Mail:
bjohnde@yahoo.co.in,
Tel: 0091- 462- 4264374,
Fax: 0091- 462-2561765


ABSTRACT
The present aim is to study the antimicrobial activity of different
plant parts of Marselia minuta L. (rhizome, frond and sporocarp)
found around the Tirunelveli, Tamilnadu. Antibacterial activity
screened against Gram +ve and Gram ve bacteria. Collected plant
materials are dried and the soluble extracts are made using organic
solvent like ethanol. Antimicrobial activities are measured using
agar cup diffusion method. Greater the area of inhibition zone
indicates the presence of good potentiality of antimicrobial activity.
Rhizome and frond extracts show good antimicrobial activity than
sporocarp.
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INTRODUCTION:
Ferns and their allies are in a major division of the Plant Kingdom called Pteridophyta, and they have
been around for millions of years. There are over 250 different genera of ferns and about 12 000
species
(1)
. It has been observed that pteridophytes are not infected by microbial pathogens, which
may be one of the important factors for the evolutionary success of pteridophytes and the fact that,
they survived for more than 350 million years
(2)
. Internationally, several researchers showed the
therapeutic properties of certain fern species. Thus, the antioxidant, antimicrobial, antiviral, anti
inflammatory, anti tussive, antitumor, anti-HIV and antihelmintic properties of various fern extracts
were demonstrated.
Marsilea minuta belongs to the family of Marseliaceae. It is a perennial fern with slender, rooted,
creeping, branching rhizomes bearing erect leaves (sterile fronds) along their length
(3)
. The leaves,
which consist of four, 'clover-like' leaflets at the apex of a slender erect stalk, arise along the length
of each rhizome. At the base of the petioles the sporocarps are formed on about 5 mm long stalks.
The sporocarps are 3-4 mm long, oblong with rounded ends (when seen laterally), with their long
axis at right angles to the stalk. It is an aquatic fern widely distributed and common fern in India with
few or no threats, hence assessed as least concern. It has the various medicinal properties. The aim of
this paper was the evaluation of antibacterial activity of crude extracts obtained from rhizome, frond
and sporocarp.
MATERIAL AND METHODS:
Fresh specimens of ferns are collected from different places of Tirunelveli, Tamilnadu. The
specimens were identified and confirmed in St. Xaviers College Herbarium, Palayamkottai. The
specimens are then shade dried. The dried specimens are powdered. Rhizome, frond and sporocarp
extracts are made from powdered specimens in organic solvent ethanol. For each specimen and for
each extraction 5 gm. powder is taken in three conical flasks (100ml) to which 20 ml solvent is
poured respectively. The mouth of flasks are tightly plugged with non-absorbent cotton and sealed
with grease to prevent evaporation. Then the flasks are placed in a shaker for about 24 hours at room
temperature37
0
C. After shaking for 24 hours, the extracts are filtered using Whatman No-1filter
paper. The filtered extracts are tested for antimicrobial activities against both gram
+ve
and gram
ve

bacteria on nutrient agar plate by disc diffusion method
(4)
. The bacterium E. coli is taken as standard
gram
ve
specimen and the bacterium Bacillus subtilis is taken as standard gram +ve specimen for
testing the antimicrobial activity.
In present experiment fresh bacterial culture solution having concentration 106 cells/ml is taken and
discs of 6mm in diameter are made on nutrient agar plate for diffusion assay. Sterile distilled water is
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used as control. After incubation for 24 hours at 37
0
C, the diameter of inhibition zones are measured
and analyzed. Three replicates are made for each set of experiment.
RESULTS AND DISCUSSION:
The result of antibacterial activity of fern Marselia minuta is shown in table 1. The results show the
well antimicrobial activity for both gram
+ve
and gram
-ve
bacteria. The rhizome and frond extracts of
this species show good antibacterial activity. It indicates that the antibacterial substances present in
rhizome and frond are in good amount. The best antimicrobial activity is found in rhizome extracts
of this species. Sporocarp extracts also show antimicrobial activity but in fewer amounts than other
extracts. Gram
+ve
bacteria shows high antibacterial activity than gram
+ve
. The pH of the extracts
are showed in table 2.
Table: 1 Antibacterial activity of Marselia minuta
S.No
Name of the
bacterium
Diameter of inhibition zone (mm)

Control
(mm)

Rhizome Frond Sporocarp

1 Escherichia coli 20.5 15.6 10.5 5.0

2. Bacillus subtilis 25.3 20.1 12.6 5.0


Table: 2 pH value of extracted samples of Marselia minuta
S.No
P
H
value of the extract

Rhizome Frond Sporocarp

1 6.2 5.9
6.0


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The frond, rhizome and rachis of ferns are covered by glands densely. These epidermal glands
(5)

contain substances like phenolic compounds, glycosides, flavonoids and alkaloids
(6,7)
. These
substances are largely responsible for the antimicrobial activity and are being soluble in organic
solvents easily extracted in ethanol but less soluble in water
(8, 9)
.
The present results show the good antimicrobial activity of species indicating the presence of good
amount substances like phenolic compounds, glycosides, flavonoids and alkaloids. These
observations are good agreement with the findings of
(10,8,)
. The antimicrobial activities of the ferns
are also in agreement with the common usage of ferns in folk medicine for bacterial infection such as
infection of throat, boil, and ulcer and in wound healing
(8)
, tumour
(11)
, dermatophytes
(12)
.
The antibiotic spectra of this fern cover both gram positive and gram negative bacteria. These
observations provide support that the ferns produce a variety of antimicrobial substances. It is
necessary to keep in mind that the factors like climatic condition, nature of plant parts, age of plant at
the time of collection etc are also responsible for the enhancement of the activity of the antimicrobial
substances and it needs to be studied more in details.
REFERENCES:
1. Chang, H.C., Gupta S.K. and Tasay H.S, 2011: Studies on Folk Medicinal Fern: An Example
of Gu-Sui-Bu in H. Fernndez, A. Kumar, M.A. Revilla (Editors), Working with Ferns,
Issues and Applications, New York, Dordrecht, Heidelberg, London, Springer, 285-304.
2. Sharma, B.D. and Vyas, M.S., 1985: Bull. of Bot. Survey of India, 27, 90-91.
3. Kostermans, A.J.G.H, Wirjahardja, S. and Dekker, R.J., 1987: The weeds: description,
ecology and control. Weeds of rice in Indonesia [edited by Soerjani, M.; Kostermans,
A.J.G.H.; Tjitrosoepomo, G. Jakarta, Indonesia; Balai Pustaka, 24-565
4. Bauer, A.W., Kirby, W.M., Sherries, J.C. and Tuck, M. 1966: Antibiotic susceptibility testing
by a standardized disc diffusion method. American Journal of Clinical Pathology, 45:493-
496.
5. Manickam, V.S., Benniamin, A. and Irudayaraj, V. 2002: Antibacterial activity of leaf
glands of Christells parasitica (L.) Lev, Indian Fern J., 22: 87-88.
6. Alcaraz, L.E, Blanco, S.E, Puig, O.N, Tomas, F, Ferretti, F.H. 2000: Antibacterial activity of
flavonoids against methicillin-resistant Staphylococcus aureus strains. J Theor Biol, 205:
231-240.
7. Cushnie, T.P.T. and Lamb, A.J. 2005: Antimicrobial activity of flavonoids. Int J Antimicrob
Agents, 26: 343-356.
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8. Adedapo, A.A., Jimoh, F.O., Koduru, S., Masika, P.J. and Afolayan, A.J. 2009: Assessment
of the medicinal potentials of the methanol extracts of the leaves and stems of Buddleja
saligna. BMC Complement Altern Med, 9: 21.
9. Banerjee, R.D. and Sen S.P. 1980: Antibiotic activity of Pteridophytes. Ecol. Bot., 34(3):
284-298.
10. Sen, S. and Nandi P. 1951: Antibiotics from the Pteridophytes. Sci. Cult., 16: 328-329.
11. Creasey,W.A.1969: Antitumoral activity of the fern Cibotium schiedei. Nature., 222: 1281-
1282.
12. Davvamani, S.N., Gowrishankar, J., Anbuganpathi, G., Srinivasan, K., Natarajan, D.,
Perumal, G., Mohanasundari, C. and Moorthy K. 2005: Studies of antimicrobial activities of
certain medicinal ferns against selected dermatophytes, Indian Fern J, 22: 191-195.
Plate: 1 Antibacterial activity of Marselia minuta