You are on page 1of 4



ISSN: 0976-9390
International Journal of
Chemical and Pharmaceutical Sciences
2013, June., Vol. 4 (2)

Antibacterial activity of Cassia auriculata linn against some pathogens
isolated from wound
Indira S* and Lakshmi M.
P.G. and Research Department of Microbiology and A Division of Biotechnology, S.T.ET. Womens College,
Sundarakkottai, Mannargudi, Thiruvarur, Tamil Nadu, India.
*Corresponding Author: E-Mail:

The in vitro antibacterial activity of Cassia auriculata L root extract of (Methanol, Ethanol ,
Aqueous) has been investigated against Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia
coli isolated from wound samples. The fresh and methanol extracts of Cassia auriculata L showed high
activity against nearly all the test microorganisms. The zone of inhibition of extract is very close and
identical in magnitude and is comparable with that of standard antibiotics used.
Key words: Cassia auriculata L, Wound, Antibacterial activity, Antibiotics.
Cassia auriculata Linn is a common plant aqueous extract of dry root of Cassia auriculata
in Asia, profoundly used in ayurvedic medicine as Linn was conducted using agar disc diffusion
a tonic, astringent and as a remedy for diabetes, method. The microorganisms used include
conjunctivitis and opthalmia. It is one of the Staphylococcus aureus, Pseudomonas aeruginosa,
principle constituents as a tonic, astringent and as Escherichia coli. The maximum activity was
a remedy for diabetes, conjunctivitis and observed against all microorganisms the
opthalmia. It is one of the principle constituents of minimum inhibitory concentration was
Avaarai panchaga chooranam- an Indian herbal determined depending on microorganisms. Cassia
formulation used in the treatment of diabetes to auriculata Linn was observed to have antibacterial
control the blood sugar value [1]. The shrub is activity and can be seed to combat against vast
especially famous for its attractive yellow flowers flora of microorganisms.
which are used in the treatment of skin disorders In an effort to expand the spectrum of
and body odour. It is widely used in traditional antibacterial agents from natural resources, Cassia
medicine for rheumatism, conjunctivitis and auriculata Linn belongs to leguminosa family has
diabetes. It has many medicinal properties. Its been selected. In the Indian literature, this plant
bark is used as an astringent, leaves and fruits has described to be useful against skin diseases,
anthelminthic, seeds used to treat in eye troubles liver troubles, tuberculosis glands and it use into
and root employed in skin disease [2]. the treatment of hematemsis, pruritis, leucoderma
Cassia auriculata Linn. commonly known and diabetics has been suggested. It has been
as tanner's cassia, also known as avaram in concluded that plant parts could be used as a
Tamil language. The shrub is especially famous for therapeutic agent in the treatment of
its attractive yellow flowers which are used in the hypercholesterolemia partially due to their fiber
treatment of skin disorders and body odour. It is and mucilage content. Besides its pharmacological
widely used in traditional medicine for uses the plant extract is also recommended as a
rheumatism, conjunctivitis and diabetes. It has pest and disease control agents in India. This plant
many medicinal properties. Its bark is used as an is widely used by tribal people to treat various
astringent, leaves and fruits anthelminthic, seeds ailments including ringworm and other fungal
used to treat in eye troubles and root employed in skin infections. The leaves are laxative and are
skin diseases. The antidiabetic, hypolipidemic and useful in skin diseases. This plant organ is known
antioxidant in skin disease and hepatoprotective to be an important source of secondary
effect [3]. metabolites, notably phenolic compounds.
Medicinal plants represent a rich source 2. EXPERIMENTAL
of antimicrobial agents. The most common 2.1. Materials
bacteria used in the pharmacognostics analyse.
Antibacterial potential of methanol, ethanol,


Research Paper

Ethanol, Methanol, Crystal violet, Grams After 24 hours, when growth was
iodine, Safranine, Ethyl alcohol, Tetracycline, observed, the colonies were sub cultured and
Amphiciline and Gentamycine were purchased were later subjected to IMVIC test.
from Madras Scientific Supplies, Trichy. 2.2.6. Collection of Plant Materials
2.2. Methodology The following medicinal plant was
2.2.1. Collection of samples selected for the study from the local area based on
their basic information available. The
The samples were collected by
(Government Hospital from Orathanadu(Tk), medicinal plant Cassia auriculata Linn root were
collected from Pinnaiyur village, Thanjavur
Thanjavur (district) using sterile cotton swab
(District) Tamil Nadu, South India, during January
from the infected patients. Sterile swab gently
rotated on the wound region. The collected
samples were immediately transferred to the 2.2.7. Preparation of root extract
laboratory for isolation of bacterial species. Cassia auriculata Linn roots were dried in
2.2.2. ISOLATION AND IDENTIFICATION OF an oven below 600 C for 2 hrs. They were finely
ORGANISMS powdered and extract with 80% water, ethanol,
methanol using Soxhlet apparatus at 55oC. The
Nutrient agar medium was poured on to
soluble part was concentrated over water bath
the sterile petriplates. After solidification, the
maintained at below 60oC and dried in a vacuum
collected wound samples were streaked on the
plate. Then the plates were incubated at 370C for metabolic extract of Cassia auriculata Linn
24 and 48 hours respectively. The isolated
colonies were identified by cultural and 2.2.8. Antibacterial activity
morphological characteristics. 2.2.9. Disc Diffusion Method
2.2.3. Identification of bacteria from wound Antibacterial activity was determined by
Samples disc diffusion method. Standard suspension of
Grams staining (Hans Christian Gram, bacteria was inoculated on the surface of Nutrient
1884) from the isolated colonies was performed agar plates. Ethanol, methanol, aequeous (1:1)
to gain the most valuable cellular and was used to dissolve the plant extract. Sterilized
morphological information about the bacteria. filter paper disc (5mm) containing 20L of each
Grams staining can be used to divide most extract (100mg/mL) was arranged on the surface
bacterial species effectively into two large groups of the inoculated plates and incubated at 370C for
such as gram positive and gram negative. It can 18-24 hours. Along with this 30g standard
also be used to examine clinical material directly Antibiotic discs(Himedia standard -Tetracycline,
[4]. Gentamycin, Ampicilin) were studied for
antimicrobial activity as a positive control
2.2.4. Motility Test
whereas the solvent used for preparing extract
Hanging drop slide with depression on was used as a negative control .At the end of
the upper side was cleaned, flamed and placed on incubation, inhibition zone formed around the
the table. A little vaseline or petroleum jelly was discs were measured with Himedia zone scale.
spread around the cavity of the slide. With the The study was performed in triplicate and the
help of match stick, petroleum jelly was applied on mean values were presented [5].
each of the four corners of the cover slip and
placed on a paper.. One loopful of culture was 2.2.10. Comparison of sensitivity of bacteria to
plant extract and antibiotics
transferred in the centre of the cover slip.
Depression slide was placed on to the cover slip; The comparative study of different
with the cavity facing down as a result the antibiotics and plant extract on bacteria were
depression covers the suspension. The slide was assayed by disc diffusion method.
gently pressed to form a seal between the cover Antibiotic discs were used to detect
slip and the slide. The preparation was lifted and antibiotic (Tetracycline, Genta mycin and
the hanging drop preparation turned quickly so Amphicillin) sensitivity of the bacterial isolates.
that the culture drop was suspended. The The nutrient agar plates were inoculated with
preparation was examined under low-power 0.1ml of bacterial suspension from nutrient broth.
objective of compound microscope with reduced The antibiotic discs were placed on the inoculated
light. Then switched to the high power objective plates and incubated at 370C for 24 hrs. Antibiotic
and preparation was examined again. sensitivity was assayed from the diameter of zone
2.2.5. Biochemical test of inhibition (Table 2, Figure 1).


Research Paper

3. RESULT AND DISCUSSION treatment of infectious diseases. Certain natural

products such as plants and different parts of
The present study is to prove the
plant leaves, roots, flowers, fruits, barks, seeds
antibacterial effect of commonly available
medicinal plant root extract against wound and oils are used to cure chronic and wound
pathogens. Our study deals with the antibacterial
causing pathogens such as Escherichia coli,
activity of Cassia auriculata Linn root extract
Pseudomonas aeroginosa and Staphylococcus
against some pathogens isolated from wound. The
aureus. Samples were collected from infected
samples were collected from infected patients by
patient by sterile cotton swab. From the
sterile cotton swab .From these samples, bacterial
collected samples, the pathogens of bacterial
species such as Escherichia coli, Pseudomonas pathogens such as Escherichia.coil, Staphylococcus
aureus Pseudomonas aeruginosa. The antibacterial
aeroginosa and Staphylococcus aureus were
potency of Cassia auriculata Linn roots were
recovered. Morphological characteristics of
tested with the three extracts (methanol, aqueous
bacterial colonies were confirmed by staining
and ethanol). From the above three extracts, the
techniques and isolated.
ethanol extracts showed the higher zone of
Organisms were also confirmed by the inhibition when compared to other by disc
biochemical activities (Table-1). The ethanol, diffusion method. Our study correlated with the
methanol and aqueous extract of Cassia auriculata methonolic extract of flowers showed significant
Linn roots were evaluated by disc diffusion reduction of blood glucose response towards
method. The ethanolic extract of C.auriculata Linn maltose ingestion and concurrently suppress
results. were showed the maximum zone of insulin activity in rats [6].
inhibition against Staphylococcus aureus 14 mm.
The aqueous and methanol extract of plant root The ethanolic extract of C. auriculata Linn
showed the maximum zone of inhibition against
extracts were showed the maximum zone of
Staphylococcus aureus (14mm). Among all the
inhibition against Escherichia coli 13 .5 and 13
extracts, the minimum activity of plant root
mm, (Table-2, and Fig-1).
extract showed at Pseudomonas aeruginosa.
Among all the extracts, the minimum Amsaveni, (2011) evaluate the antibacterial
activity of C.auriculata Linn roots showed at efficacy of Cassia auriculata Linn against Extented
Pseudomonas aeruginosa(8mm). The use of plant Spectrum Beta Lactamase (ESBL) produced E. Coli
compounds for pharmaceutical purpose has Methanol, ethanol ethyl acetate, hexane and
gradually increased. Several synthetic antibiotics chloroform crude and Soxhlet extracts of ethyl
are employed in the treatment of infections and acetate and hexane of leaves, roots and flowers of
communicable diseases. The harmful Cassia auriculata Linn were tested for
microorganism can be controlled with drugs and antibacterial activity by well diffusion method
the presence of multiple drug resistant bacteria against ESBL produced Escherichia coli.
created alarming clinical situations in the
Table - 1: Identification of bacterial Isolates by Biochemical test
Name of The Organisms
Name of the test Escherichia Coli Staphylococcus aureus Pseudomonas earuginosa
Gram staining + + +
Indole test + - -
Methyl red test + + -
Vogasproskauer - + -
Citrate utilization - - +
Catalase test + + +
Triple sugar Iron + - +
Table - 2: Antibacterial activity of Wound isolates
Organisms Cassia auriculata L Tetracycline Gentamycin Amphicilin
Escherichia coli 14.49.6 12.444.6 11.90.57 10.80.08
Staphylococcus aureus 12.05.06 10.30.05 10.90.00 9.90.00
Pseudomonas aeruginosa 11.20.01 9.70.5 9.05.88 9.72.5


Research Paper

Earlier study of ethanolic and methanolic 2. Kumar RS, Ponmozhi M, Viswanathan P and
extracts of the flowers showed antioxidant activity Nalini N. J. of Nutritional Biochemistry,
which may due to presence of flavonoids and 2003; 14: 452-458.
tannins. The aqueous and ethanolic extract of 3. Cappuccino and Nataire Shreeman S.
flowers at dose 0.25 and 0.5 kg for 30 days Microbiol., 1999; 28: 190-194.
exhibited a significant reduction in blood glucose,
serum triglycerides, total serum cholesterol level 4. Abesundara R, Muthutheyaru K, Umeshankar
and remarkable increase in plasma insulin level G, Muralitharan G, Cordairayen E and
[7]. The combined aqueous extract of C.auriclata L. Vasantha. Ind. J. Pharm Sci, 2004; 66(1):123-
and Aegle marmelos doses of 250,350 and 450 125.
mg showed a significant reduction in glucose and 5. Kumaran and Karunakaran JN. J.
lipid in serum on rats in dose dependent manner. Ethnopharmacol., 2007; 60:1-5.
It is possessed increment in serum insulin and
restoration of beta cells [8]. It is clearly evident 6. Praveen Kumar A and Randhir Singh Dahiya S.
through this study that synthetic drugs can be Biomedicine, 2012; 28: 190-194.
replaced with natural medicine (herbal medicine) 7. Sabu MC and Subburaju T. J.
which is safer to human systems and also friendly Ethnopharmacol., 2007; 80: 203-206.
to human body. More research is related to design
8. Sivaraj K, Jansen AM, Cheffer JJC and Svendsen
and pattern this medicine of herbal drugs [9].
AB. Planta. Med., 2009; 40:395-398.
9. Yesu Raj, D Arcy PF and Griffin JP. Adverse
1. Kumar RS, Ponmozhi M and Nalini M. Asia Drug React Toxicol. Rev., 2012; 13(2):65-76.
Pacific J. of Clinical Nutrition, 2002; 11: 2012.