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Ethnobotanical
Leaflets 12: 728-33. 2008. In-Vitro
Antimicrobial Activity of Extracts of Passiflora edulis (Passifloraceae) and Sphaeranthus indicus (Asteraceae) *Asirvathamdoss, Pichai Anthoni Doss and
Rangasamydhanabalan Department of Microbiology,
RVS *Corresponding author: dossandro@gmail.com Issued ABSTRACT ����������� The
organic extracts of Passiflora
edulis and Sphaeranthus
indicus were evaluated for antimicrobial activity against clinically
important bacteria viz., Escherichia coli MTCC 443, Salmonella
typhi MTCC 734, Staphylococus aureus MTCC 737, Bacillus
subtilis, Streptococcus pyogens, Pseudomonas aeruginosa, Klebsiella pneumoniae,
Proteus vulgaris, Aspergillus flavus, Pencillium restrictum and Trichoderma
viride. The in-vitro antimicrobial activity was performed by agar disc
diffusion method. Streptococcus pyogens was the most resistant
bacterial strains against the methanol extracts. Key
words: Antimicrobial
activity, Petroleum ether, Methanol, Benzene, Chloroform, Ethyl acetate, hot
water Passiflora edulis and Sphaeranthus indicus INTRODUCTION ����������� Nature
has been a source of medicinal agents for thousands of years and an impressive
number of modern drugs have been isolated from natural sources, many based on
their use in traditional medicine (.R. Nair, et al.,2005) . Plants have a
great potential for producing new drugs of great benefit to mankind
(PAREKH et al., 2005). During last ten years the pace of development of new antimicrobial
drugs has slowed down while the prevalence of resistance bacteria is no
longer matched by expansion in the arsenal of agents (D.A. Akinpelu and T.M. Onakoya ., 2006). Many efforts have been
done to discover new antimicrobial compounds from various kinds of sources
such as soil, microorganisms, animals and plants. One of such resources is
folk medicine and systematic screening of them may result in the discovery of
novel effective compounds ((Janovska et al., 2003). The need of the hour is
to screen a number of medicinal plants for promising biological activity.
There are several reports on the antimicrobial activity of different herbal
extracts in different regions of the world (Chung et al., 2004; Nair R,
Chanda SV., 2004; De Boer HJ et al., 2005; Nair R et al., 2005). Because of
the side effects and the resistance that pathogenic microorganisms build
against antibiotics, recently much attention has been paid to extracts and
biologically active compounds isolated from plant species used in herbal
medicine (Essawi T, Srour M .,2000). Plant-based antimicrobials represent a
vast untapped source of medicines and further exploration of plant
antimicrobials needs to occur. Considering the aforesaid, the three
traditionally used medicinal plants belonging to three different families
were screened for their antimicrobial properties. Passiflora edulis (Passifloraceae)and Sphaeranthus indicus (Asteraceae). Passiflora
edulis (climber) Sims belongs to Passifloraceae. It is used in homeopathic medicine for the
treatment of insomnia, epilepsy, tetanus, muscle spasms and leaves presented
anxiolytic activity (L.N. Yuldashevaet al.,2005).Sphaeranthus indicus Linn
(herb) belongs to family Asteraceae . The plant is distributed
through out the plains and wet lands in MATERIAL
Plant material ����������� Fresh
plant samples were collected from The Nilgiris, Preparation of extracts: ����������� For solvent extraction, 50 g of air-dried powder was taken in 250 ml of organic solvent (Petroleum ether, chloroform, benzene, ethyl acetate methanol and hot water) in a conical flask, plugged with cotton wool and then kept on a rotary shaker at 190-220 rpm for 24 h. After 24 hours the supernatant was collected and the solvent was evaporated at room temperature. The repeated procedure was followed all the other solvents and water. The extracts were stored at 4 0C in airtight bottles. Microorganisms: ����������� �In vitro antimicrobial activity
was examined for all the organic extracts from two medicinal plants used by traditional
healers. Escherichia coli MTCC 443,
Salmonella typhi MTCC 734, Staphylococus aureus MTCC 737 was
obtained from IMTECH, Phytochemical Screening Test Phytochemical screening is done for analyzing secondary metabolites, which are responsible for curing ailment. The phytochemical screening of the plant extract was carried out by following methods used by Amarasingham et al.,(1964),Das and Bhattacharjee, (1970),Gibbs, (1974), Trease and Evans (1978), Santaram and Harborne (1984) to detect the presence or absence of certain bioactive compounds. Antibacterial
Assay
����������� Plant extracts of P.edulis and S.indicus, which was prepared with different solvents like Petroleum ether, Benzene, Chloroform, Ethyl acetate, Methanol and Water were used to test their antibacterial activity. ����������� Antibacterial activity
was demonstrated using a modification of the method originally described by
Bauer et al (1966) which is widely used for the antibacterial
susceptibility testing .Liquid nutrient agar media and the Petri plates were
sterilized by autoclaving at 120o C for 30 minutes. Under aseptic
conditions in the laminar airflow chamber, about 20ml of the agar medium was
dispensed into each Petri plate to yield a uniform depth of 4mm. After
solidification of the media the bacterial strains were swabbed on the surface
of the agar plates. ����������� Whatmann No.1 filter paper
was cut into small discs of diameter 0.4cm and autoclaved. These discs were
dipped into the different plant extracts of each three concentrations namely
(20mg/ml, 10mg/ml, 5mg/ml). The dipped discs were placed on the appropriate
swabbed Petri plates such that each Petri plates have three concentration of
each plant extract.����������� It was
then incubated at 37o C for 24 hours the antibacterial drug which
was used here was chloramphenicol for positive control and DMSO was used for
negative control. After incubation the zone of inhibition was measured in mm.
RESULTS ����������� The data reported in Table 1 presents the antimicrobial activity of the organic extracts of P.edulis and S.indicus. The results indicate that the extracts from the medicinal plants studied showed inhibition of growth of some of the tested micro organisms with to various degrees. The antibacterial activity of S. indicus and P.edulis leaf extract of all the solvents against Escherichia coli MTCC 443, Salmonella typhi MTCC 734, Staphylococus aureus MTCC 737, Bacillus subtilis, Streptococcus pyogens, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus vulgaris, Aspergillus flavus, Pencillium restrictum and Trichoderma viride was evaluated . The methanolic extract of both plants (P.edulis and S.indicus ) showed considerably more activity when compared to other all the solvents. Maximum antibacterial activity was shown against Streptococcus pyrogens, followed by S. aureus and B.subtilis. From the above results it can be concluded that plant extracts have great potential as antimicrobial compounds against microorganisms and that they can be used in the treatment of infectious diseases caused by resistant microorganisms. S. indicus showed maximum antibacterial activity and so this plant can be used to discover bioactive natural products that may serve as leads for the development of new pharmaceuticals that address hither to unmet therapeutic needs. Such screening of various natural organic compounds and identifying active agents is the need of the hour, because successful prediction of lead molecule and drug like properties at the onset of drug discovery will pay off later in drug development. REFERENCES � Ambavade SD, Mhetre NA, Tate VD, Bodhankar SL.,2006. Pharmacological evaluation of the extracts of Sphaeranthus indicus flowers on anxiolytic activity in mice.,Indian Journal of Pharmacology.,38 (4) :254-259 � R. Nair, T. Kalariya, Sumitra Chanda.,2005. Antibacterial Activity of Some Selected Indian Medicinal Flora, Turk J Biol.,29 : 41-47 �
Jigna Parekh, and Sumitra Chanda.,2006. In-vitro Antimicrobial Activities of Extracts of Launaea procumbens Roxb.
(Labiateae), Vitis vinifera L. (Vitaceae) and Cyperus rotundus L.
(Cyperaceae), African Journal of Biomedical
Research., 9 ; 89 -93 � Janovska, D., Kubikova, K., Kokoska, L. (2003).Screening for antimicrobial activity of some medicinal plant species of traditional Chinese medicine. Czech. J.Food Sci. 21: 107-111. � D.A. Akinpelu1 and T.M. Onakoya2.,2006. Antimicrobial activities of medicinal plants used in folklore remedies in south-western. African Journal of Biotechnology ., 5 (11): 1078-1081 � Jigna Parekh, Darshana Jadeja, Sumitra Chanda.,2005.Efficacy of Aqueous and Methanol Extracts of Some Medicinal Plants for Potential Antibacterial Activity., Turk J Biol.,29 : 203-210 � Chung PY, Chung LY, Ngeow YF et al.,2004. Antimicrobial activities of Malaysian plant species. Pharm Biol 42: 292-300. � Nair R, Chanda SV.,2004. Antibacterial activity of some medicinal plants of Saurashtra region. J Tissue Res 4: 117-120. �
De Boer HJ, Kool A, Broberg A et al.,2005.
Antifungal and antibacterial activity of some herbal remedies from � Nair R, Kalariya T, Chanda S.,2005. Antibacterial activity of some selected Indian medicinal flora. Turk J Biol 29: 41-47. � Essawi T, Srour M.,2000. Screening of some Palestinian medicinal plants for antibacterial activity. J Ethnopharmacol 70: 343-349. � L.N. Yuldasheva, E.B. Carvalho, M.-T.J.A. Catanho and O.V. Krasilnikov .,2005. Cholesterol-dependent hemolytic activity of Passiflora quadrangularis leaves., 38(7) 1061-1070 �
Amarasingam, R.P., Bisset, N.G., Millard, A.K.
and Woods, M.C, (1964). Phytochemical survey of � Bauer, A.W., Kirby, W.M.M.Sheeis, Durk M., (1996).Antibiotc susceptibility testing by a standard single disc method. Amer.j.chin Pathol.36: 493-496. � Das, A.K. and Bhatacharjee, A.K, (1970). A systematic approach to Phytochemical Screening, Trop, Sci.1254-58. �
Gibbs, R.D, (1974), Chemotaxonomy of
flowering plants. �
Trease. G.S., and Evans, H.C, (1978). Textbook
of Pharmacognosy. 9 th edition. Bailiar Zindall And Co., �
Harborne, J.B, (1984) (Eds.) Phytochemical
method a Chaman and Hall, Table 1. Phytochemical screening of Sphaeranthus indicus.
Table 2. Phytochemical screening of Passiflora
edulis (leaves).
�
�+�
- Positive������������������������ �-� -
Negative TP
��������� �-������������� Tannin �& Phenolic compound�������������������������������������� FL
��������� �-������������� Flavonoids
�������������������������� � ST
��������� �-������������� Steroids TE
��������� �-������������� Terpenoid������������� AA
������� �-������������� Amino
acids������������������������� GL ��������� �-������������� Glycosides OF ��������� �-������������� Oil& Fat���������������� ���� �����������CG����������
-������������ �Cardioglycosides CH
�������� �-������������� Carbohydrates Table
1a. Antibacterial activity of Sphaeranthus
indicus (Whole plant).
Table
1b. Antibacterial activity of Passiflora edulis (Leaves).
�
*Zone of inhibition in mm (Test were done in
duplicate) Standard
B.s � Bacillus subtilis , S.a - Staphylococcus aureus ,
S.p-Streptococcus pyrogens, E. coli �Eschershia coli , S.t � Salmonella typhi
, K.p-Klebsiellae pneumoniae, P.a-Pseudomonas aeruginosa |