investigate in vitro anti microbial activity of aerial parts of Calotropis
procera and Centella asatica Linn.,
that have been popularly used as folk medicines. The organic solvent plant
extracts are tested on the various micro-organisms including bacteria and fungi
using agar well diffusion technique. The length of the inhibition zone was
measured in millimeters from the edge of the well to the edge of the inhibition
zone. Calotropis procera showed significant to
moderate activity against (14mm) Pseudomonas marginalis
and (21mm) Streptococcus mutans with 100mg/ml DMSO
plant drug concentration. The results of (MICs) values are lowest at 66 and
highest at 152mg/ml for Calotropis asatica. The
extracts were assessed in an effort to validate the potential activity of the
medicinal plant plants against microbes.
of Content vi-vii
1.0 Introduction 1-6
2.0 Literature Review 7
2.1 Local Names 7
2.2 Botanical Description 7-8
2.3 Biology 8
2.4 Ecology 8
2.5 Biophysical Limits 8
2.6 Documented Species Distribution 8-9
2.7 Exotic 9
2.8 Products 9-11
3.0 Materials and Methods 12
3.1 Plant Materials and Extraction 12-13
3.2 Determination of Antimicrobial Activity 13-15
3.4 Results and Discussion 15-20
4.0 Result and Discussion 21
4.1 Proximate Analysis 21
4.2 Determination of Moisture Content 21
4.3 Determination of Crude Protein 21-22
4.4 Determination of Ash Content 22-23
4.5 Determination of Fat Content 23-24
4.6 Determination of the Crude Fiber 24-25
4.7 Determination of Carbohydrate or Nitrogen
Free Extract 25-27
5.0 Recommendation and Conclusion 28
5.1 Recommendation 28
5.2 Conclusion 28
According to a study conducted by
the World Health Organization (WHO) based on publications on pharmacopoeias and
medicinal plants in 91 countries, the number of medicinal plants is nearly 21,000.
Nearly 6-7 thousand species of medicinal plants out of about 17-18 thousand
flowering plants are known to be in use in folla and officially recognized
systems of medicine in India i.e. Ayurbeda, Sidha, Unani and Homeopathy.
India has a rich heritage of
knowledge on plant based, drugs both for use in preventive and curative
medicine. From ancient civilization the various parts of different plants were
used to eliminate pain, control suffering and counteract diseases. Plants
generally produce many secondary metabolites which constitute an important
source of microbicides, pesticides and many pharmaceutical drugs. It has also
been widely observed and accepted that the medicinal value of plants lies in
the bioactive phytocomponents present in the plants (Veeramuthu et al;
2006, Khan and Khan, 2008, Ghosh et al; 2008, Mandal et al; 2010,
Abubakar et al; 2011). Much work has been done on ethnomedicinal plant
in india (Nair 2005; Rout et al; 2009, Madhuri Sharma and Pandey Govind,
2009). Medicinal plants represent a rich source of antimicrobial agents (Kannan
et al; 2007); Jeeshna et al; Jarrar et al; 2010, Johnson et
al; 2010, Koochak et al; 2010). Because of the side effects and the
resistance that pathogenic microorganisms build against antibiotics, many
scientists have recently paid attention to extracts and biologically active
compounds isolated from plant species used in herbal medicine (Mst Nazma Yesmin
et al; 2008).
Calotropis procera linn. (Family Asclepiadeceae) is
a shrub or small tree up to 2.5 m height. All parts of the plant exude while
latex when cut or broken. It is known by various names like swallow wort, dead sea
apple, Sodom apple or milk weed, commonly used and known as Arka or madar.
Telugu name is Jilledachetta and in English Catotrope, Calotropis, Deadsea
fruit, desert wick, giant milk weed, muder fibre, rubber bush, rubber tree,
Sodom apple, swallow, wort. In the traditional Indian medicinal system, it has
been used for pain, asthma, bronchitis and dyspepsia, leprosy, Ulcers, tumors,
piles, spleen, liver and abdomen. The root bark in the form of paste with rice
vinegar is applied to elephantiasis of the legs and scrotum. The plant is also
known for its toxic properties that include iridocyclitis, dermatitis and acts
like a poison and produces lethal effects. Compounds derived from the plant
have been found to have emeto, cathartic and digitalic properties.
Antimicrobial activity of C. procera was previously
screened by (Kareem et al; 2008; Verahalarao Vadlapudi et al;
2004). In the leaves, mudarine is the principal active constituent as well as a
bitter yellow acid, resin and 3 toxic glycosides calotropin, Uscharin and
Calotoxin. The latex contains a powerful bacteriological enzyme, a very toxic
glycosides calactin (the concentration of which is increased following insect
or grasshopper attack as defense mechanism), Catatropin D1, Calotropin DII,
Calotropin F1, Calotropin F11, and a non toxic protealytic enzyme calotropin (2
% to 3 %). This calotropin is more proteolytic than papain and bromelain
coagulates milk and digest meat, gelatin and casein. The whole plant contains a
b-amyrin, teraxasterol, gigcintin, giganteol, isogiganteol, b-sitosterol and a
Centella asiatica (Family Mackinlayaceae) common
name include Gotukola, Asiatic pennywort, Indian Pennywort, Luei Gong Gen,
Takip, Kohol, Antanan, Pegagan, Pegaga Vallaarai leula kud, Baibua bok and
Brahmi. In India, famous as “Brahm”, it is used for improving the mental
ability as was carried out at Dr. A Laleshmipathy research center (now under
CCRAS) VHS, Adyar, Chennai (Appa Rao et al; 1973). Anti bacterial
activity of C. asiatica was previously screened by (Rohimi
Kiran Kunta et al; 2004).
The aim of the present study is
to investigate in vitro antimicrobial activity of Indian medicinal plants Calotrophs
procera and Calotrophs Asatica linn
against the resistant pathogens.
Antimicrobial activity of methanol extracts of Calotrophs procera
2: Antimicrobial activity of methanol extracts of Calotrophs asiatica
Other pathogens F.
Oxysporum, L. acidophilus, S. saliuarious
and S. aureus with 100mg/ml DMSO.
The results of MICs
values are lowest at 66 and highest at 152 mg/ml for Calotrophs procera
whereas 0.155mg/ml for Calotropis asatica. The variation
of antimicrobial activity of our extracts. No variation of antimicrobial
activity of our extracts might be due to the distribution of anti microbial
substances which varied from fraction to fraction of the crude extract. No
inhibitions were observed were observed with Calotropis procera
on P. expansum and Calotrophs asatica
on P. aeruginosa and S. mutans.
These extracts are
harmless and nonphytotoxic, it has been proved that extracts have inhibitory
effects on germination and on the viability of fungal spores as well. Both
plant extracts showed moderate to good activity against A. niger as it is a
saprophyte in soil causing black mould of Onion, garlic and Shallot, stem rod
of Dracaena, root stalla rot of Sanseviera, and boll rot of Cotton; spoilage of
Cashew kernels, dates, Figs. Vanilla pods and dried prune.
The effectiveness of
the active compounds present in plant extracts causes the production of growth
inhibition zones that appear as clear as surrounding the wells. However, plant
extracts was unable to exhibit antibacterial activity against tested bacterial
strains. These bacterial strains may have some kind of resistance mechanisms
e.g enzymatic inactivation, target sits modification and decrease intracellular
drug accumulation (Schwarz and Noble, 1999) or the concentration of the
compound used may not be sufficient.
The adverse effect of
Calotropis procera consumption are reported to cause
blisters, lesions and eruptions when taken by patients for the treatment of
joint pains and gastrointestinal problems. Due to its toxicity, the latex
extracted from the stem has traditionally been used to make poison arroles. The
latex is highly toxic to human eyes and produces sudden painless dimness of
vision with photophobia (Basak et al; 2009).
phytochemicals are identified in different parts. Calotropis procera
flowers contain terpenes, multiflorenol, and cyclisadol (Al-yahya et al;
1990). The latex contains caoutcholic calotropin, calotoxin 0.15%, Calactin
0.15%, Uscharin 0.45%, Trypsin, Voruscharin, Uzarigenin, Syriogenin and
proceroside (Atef et al; 1999). Chemical constituents of calotrophs
procera flowers are lupeol, Uscharin, proceroside, proceragenin (Cardenolide),
Syriogenin, Taraxase-20 (30)-en-3-(4-methyl-3-periotenoate), 3-thiazoline
cardenolide, gigantin, giganteol, isogiganteol, Uscharidin, Uzarigenin
Voruscharin a, Calotropeol, 3-epimoretenol, a-lactuceryl acetate and
alactuceryl isovalerate (Ansari and Ali, 199). Root bark of Calotropis
procera contains triterpenes, A new norditerpenyl ester, name
calotropterpenyl ester, and two unknown pentacyclic triterpenoids, namely
Calotropursenyl acetate and Calotropfriedelenyl acetate, akundarol
isovaleratae, mundarol isovalerate and quercetin, 3-rutinoside (Akhtar and
Malik, 1998; Ansari et al, 2001). The principal active medicinals are
asclepin and mudarin (Raghubir et al; 1999).
No inhibition was
observed with controls, which proves that solvents could not act as
antimicrobial agents. In almost all test, crude methanolic extracts showed
better inhibition against all the tested bacterial and fungal strains,
indicating that active ingredients in plant materials could be extracted into
However, the highest
antibacterial activity of Calotropis procera was observed
due to the presence of secondary metabolites such as alkaloids, flavanoids and
steroids against S. aureus.
P. aeruginosa are
wide spread in soil, water and sewage and this can be considered as an
indication of their involvement in the natural process of mineralization of
organic matter. It has long been a troublesome cause of secondary infections of
wound, especially burns, giving rise to blue green pus. It produces meningitis,
when introduced by lumber puncture and urinary tract infection when introduced
by Catheters and instruments or irrigation solutions (Willey et al;
S. aureus occur
harmlessly as a normal flora of the skin and mucous membrane and it is one of
the commonest bacterial pathogens encountered in the community causing severe
food poisoning or minor skin infections to several life threatening infections
(Brooks et al; 2004).
methanol extract having strong inhibition activity against P. aeruginosa and S.
aureus was previously reported by (Thangavel Arumugam et al; 2001)
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