ABSTRACT- Medicinal Plants have been practiced for hundreds of centuries by tribes all over the world. From the earliest times until the end of nineteenth century plants are still the common source of medicinal treatment yet. Using natural, plant-derived medicines that are “healthier” then prescription drugs derived from synthesized products is something that appeals to consumers. The medicinal plants are of great importance because there are utilized as medicines. Aim of this research work was to evaluate the antibacterial activity of Skimmia laureola plant against various patho-genic strains of bacteria. The hot and cold water extract of Skimmia laureola were used against four bacterial strains Escherichia coli,Bacillus subti-lus, Staphylococcusaureus and Proteus mirabilis in order to check the antibacterial activity of Skimmia laureola. Antibacterial activity was conducted by agar well diffusion method. The Skimmia laureola showed different level of antibacterial activity. The hot and cold water extract of Skimmia lau-reola showed antibacterial activity against the micro-organism but not too maximum. Keywords: Medicinal Plants, Skimmia Laureola, Antibacterial Activity.

1. International Journal of Life-Sciences Scientific Research (IJLSSR), VOLUME 1, ISSUE 1, pp: 33-36 SEPTEMBER-2015
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ANTIBACTERIALACTIVITY OF SKIMMIA LAUREOLA
Muhammad Aurang Zeb1
, Abdul Halim1*
, Salim Ullah1
, Najeeb Ullah1
, Saadat Ullah Khan1
, M.Salahuddin2
, Mahjabin Rashid3
1
Department of Biochemistry, Hazara University Mansehra, Khyber Pakhtunkhwa, Pakistan
2
Faculty of Medicine, University of Hong Kong, Pokfulam, Hong Kong
3
College of Medicine, Mymensingh Medical College, University of Dhaka, Bangladesh
ABSTRACT- Medicinal Plants have been practiced for hundreds of centuries by tribes all over the world. From the earliest times until the end of
nineteenth century plants are still the common source of medicinal treatment yet. Using natural, plant-derived medicines that are “healthier” then
prescription drugs derived from synthesized products is something that appeals to consumers. The medicinal plants are of great importance because
there are utilized as medicines. Aim of this research work was to evaluate the antibacterial activity of Skimmia laureola plant against various patho-
genic strains of bacteria. The hot and cold water extract of Skimmia laureola were used against four bacterial strains Escherichia coli,Bacillus subti-
lus, Staphylococcusaureus and Proteus mirabilis in order to check the antibacterial activity of Skimmia laureola. Antibacterial activity was conducted
by agar well diffusion method. The Skimmia laureola showed different level of antibacterial activity. The hot and cold water extract of Skimmia lau-
reola showed antibacterial activity against the micro-organism but not too maximum.
Keywords: Medicinal Plants, Skimmia Laureola, Antibacterial Activity.
-------------------------------------------------IJLSSR-----------------------------------------------
INTRODUCTION
A medicinal plant is any plant which, in one or more of its organ, con-
tains substance that can be used for therapeutic purpose or which is a
precursor for synthesis of useful drugs.”(Sofowora, 1982).This definition
of medicinal plant has been formulated by WHO (World Health Organi-
zation). The plants that possess therapeutic properties or exert beneficial
pharmacological effects on the animal body are generally designated as
“Medicinal Plants”. It has now been established that the plants which
naturally synthesis and accumulate some secondary metabolites, like
alkaloids, glycosides, tannins, volatiles oils and contain minerals and
vitamins, possess medicinal properties. Medicinal plants are very impor-
tant for the cure of different microbial infections (Pennanen et al. 1996).
Plants have been utilized as medicines for thousands of years. These
medicines initially took the form of crude drugs such as tinctures, teas,
poultices, powders, and others herbal formulations. There are 4,22,127
plant species growing on planet earth about 35,000 to 70,000 plants spe-
cies are used as medicinal plants (Hasan et al. 2007).
Received: 03 August 2015/Revised: 14 August 2015/Accepted: 26 August 2015
Out of which 20,000 plants species are believed to be used medicinally
in the third world (Mukherjee 2004). Approximately 6000 species of
flowering plants occur in Pakistan and 700 of them have medicinal value
(Stewart 1972). With an estimation of WHO that as many as 80% of
world’s population living in rural areas rely on herbal traditional medi-
cines as their primary health care, the study on properties and uses of
medicinal plants are getting growing interests. In recent years this inter-
est to evaluate plants possessing antibacterial activity for various diseas-
es is growing (Clark and Hufford 1993). The screening of plant extracts
and plant products for antimicrobial activity has shown that higher plants
represent a potential source of new anti-infective agents (Smet 1997,
Cowan 1999,). The leaves, flowers, fruits and roots are extensively used
for treating cold, cough, whooping-cough (Dhuley 1999). Plant extracts
have great potential as antimicrobial compounds, especially in the treat-
ment of infectious diseases caused by resistant micro-organisms (Nasir
and Chanda 2006). It is important to mention that over 75% of popula-
tion in Pakistan is cured by using traditional medicines prescribed by
more than 50,000 traditional herb practitioners.
The different system of eastern medicines that is, Unani, Ayurvedic and
homeopathy etc are entirely based on medicinal properties of these
plants. The practice of traditional medicine is widespread in China, In-
dia, Japan, Pakistan, Sri Lanka and Thailand (Hasan et al. 2007). Today
herbal products and extracts are widely used to control various human
diseases (Srinivasan et al. 2006). The main objective of the research is to
screen and evaluate antibacterial activity of crude ethanol extract and to
find out minimum bactericidal concentration (MBC) against these ex-
*
Address for Correspondence:
Abdul Halim
Department of Biochemistry
Hazara University Mansehra
Khyber Pakhtunkhwa, Pakistan.
Email: abdulhalimbiochemist@gmail.com
Research Article (Open access)

2. International Journal of Life-sciences Scientific Research (IJLSSR), VOLUME 1, ISSUE 1
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tracts both gram positive as well as gram negative bacteria.
MATERIAL AND METHODS
Collection of sample
Skimmia laureola was obtained from Pansar Store near Lari Adaa, Man-
sehra (Figure 1). The plant was identified by the Department of Botany
and Herbarium, Hazara University Mansehra, Khyber Pakhtunkhwa,
Pakistan.
Figure 1: Skimmia laureola
Selection of media
During the whole research project two types of media were used that is
Nutrient Agar and Simple Agar. Nutrient Agar is the best culturing media
for testing micro-organism because it provide nutrient for the growth of
all type of bacteria.20 g of nutrient agar and 4g of simple agar was taken
for the preparation of media.
a) Composition of nutrient and simple agar
Table: 1 Composition of nutrient agar
Serial
No.
Components
1 Yeast extract
2 Beef extract
3 Peptones
4 Glucose monohydrates
5 Agar
6 Sodium chloride
Table: 2 Composition of simple agar
Serial
No
Components
1 Agarose
2 Agaropectin
Preparation of media
20 gram of nutrient agar was dissolved in 1 liter of distilled water in a
conical flask and 4 grams of simple agar is also added and plugged in
flask and shackedto mix well. Then it is heated on the hot plate stirrer to
dissolve the media completely. The media and all glass ware swabs were
sterilize by means of autoclaving under the 15psi and 121 degree centi-
grade temperature for 15 minutes in autoclave. After this media was
poured aseptically into Petri dishes in laminar flow cabinet.
Preparation of plant extract
For the attainment of maximum plant antimicrobial activity agar well
diffusion method was used. The plant was dried at sunlight and then
plant were crushed into coarse powder using grinder and then stored in
clean, dried plastic bags for further processing From powder about 20
grams was taken for experimental use to prepare plant extracts (Figure 2)
Figure 2: Powder form of Skimmia laureola
a) Cold water extraction
10 grams powder of Skimmia laureola plant was soaked in cold 100 ml
distilled water and shaken it on electric rotator at 200 rpm for 24 hours.
After 24 hours the solution was filtered through a filter paper then centri-
fuged at 4400 rpm for 7 minutes and repeat it 3 times the supernatant
appeared at top was collected which was considered as 100% pure plant
extract while the pellet appeared at bottom of centrifuge tubes was dis-
carded the pure extract were then ready for antimicrobial sensitivity test.
(Figure 3, 4 & 5).
Electric shaker Centrifuge 5702 Cold water extraction of
Skimmia laureola
Figure: 3, 4 & 5 Electric shaker and Centrifuge &Cold water extrac-
tion of Skimmia laureola
b) Hot water extraction
10 gram of Skimmia laureola powder was soaked in 100 ml distilled
water in a conical flask and then it placed in the incubator at 37degree
centigrade for 12 hours after this it was placed in the hot water bath for 2
hours and then centrifuge it for 7 minutes at 4400 rpm then filtered it

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through filter paper and repeated this process for 3 times the supernatant
obtained from 3rd
time centrifugation collected while pellet were dis-
carded and this was considered as a pure hot extract of these plants and
these were ready for sensitivity tests. (Figure 6, 7 & 8 ).
Electric shaker Centrifuge 5702 Hot water extraction
of Skimmia laureola
Figure: 6, 7 & 8 Electric shaker and Centrifuge & Hot water extrac-
tion of Skimmia laureola
Antimicrobial Assay
Media which was prepared and autoclaved was smeared or spread on the
Petri dishes in laminar flow cabinet. The Electric fan of laminar flow
cabinet was turned on to solidify the media and the pores are made in
Petri dishes containing media by tips in laminar flow cabinet. Then the
sterilized cotton swab was dipped in the distilled water and then dipped
in the bacterial culture placed it on the Petri dish containing media in
order to streak culture on the surface of nutrient agar media of Petri dish
uniformly. One cotton swab is used for only once streaking of one Petri
dish then discarded (cotton swab). Poured the hot and cold water extracts
of plants in the well in media of Petri dish by micro pipette of 100 ml.
After pouring all plates or Petri dishes were incubated in electric oven or
incubator for about 24 hours at 37 degree centigrade. And then antibac-
terial activity was checked. The zone of inhibition was measured by
scale in mm after 24 hours the antibacterial activity were assigned ac-
cording to the zone of inhibition produced by the plant extracts.
RESULTS AND DISCUSSION
Results
In the present research project the antibacterial activity of Skimmia lau-
reola, were checked against four bacterial strains among which were
Gram positive and were Gram negative. These strains included Staphylo-
coccus aureus, E.coli, Proteus mirabilis and Bacillus subtilis. The hot
and cold water extract of Skimmia laureola, were checked against the
four bacterial strains. The inhibitory activity which were shown by cold
and hot water extract against the micro-organisms.
Table: 2 Hot water extraction of Skimmia leureola.
S.No Micro-
Organism
Zone of inhibition in mm Mean
1 E.coli 11mm 13mm 11mm 11.6mm
2 B. subtilis 13mm 15mm 14mm 14mm
3 S. aureus 20mm 20mm 19mm 19.6mm
4 P. mirabilis 0mm 0mm 0mm Nil
Table 3: Cold water extraction of Skimmia leureola
S.No Micro-
Organism
Zone of inhibition in mm Mean
1 E. coli 0mm 0mm 0mm Nil
2 B. subtilis 10mm 9mm 11mm 10mm
3 S. aureus 11mm 10mm 12mm 11mm
4 P. mirabilis 10mm 10mm 9mm 9.66mm
DISCUSSION
Plants are important source of potentially useful structures for the devel-
opment of new chemotherapeutic agents. The first step towards this goal
is the in vitro antibacterial activity assay (Tona et al. 1998). Many reports
are available on the antiviral, antibacterial, antifungal, anthelmintic,
antimolluscal and anti-inflammator properties of plants. Some of these
observations have helped in identifying the active principle responsible
for such activities and in the developing drugs for the therapeutic use in
human beings. According to literature survey that Skimmia laureola was
found to have antibacterial activity the Proteus mirabilis and Salmonella
typhi at a concentration of 200 micro grams. Ampicillin, Tobramycin and
Amoxicillin were used as standard drugs. In the current investigation the
antibacterial activity of Skimmia laureola were checked against four
pathogenic bacterial strains among them two were Gram positive such as
Staphylococcus aureus and Proteus mirabilis and two were Gram nega-
tive such as Escherichia coli and Bacillus subtilis. The cold water extract
of Skimmia laureola showed the moderate antibacterial activity against
the Bacillus subtilis, Staphylococcus aureus and Proteus mirabilis. The
zone of inhibition measured were 10mm, 11mm and 9.66mm respective-
ly but did not show any antibacterial activity against Escherichia coli.
The hot water extract of Skimmia laureola showed the antibacterial ac-
tivity against the Staphylococcus aureus the zone of inhibition measured
were 19.6mm and showed a moderate activity against Eschrichia coli
and Bacillus subtilis but did not show any antibacterial activity Proteus
mirabilis.
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