ANTIBACTERIAL ACTIVITY OF SKIMMIA LAUREOLA

International Journal of Life-Sciences Scientific Research (IJLSSR), VOLUME 1, ISSUE 1, pp: 33-36 SEPTEMBER-2015

http://ijlssr.com IJLSSR © 2015 All rights are reserved

ANTIBACTERIAL ACTIVITY OF SKIMMIA LAUREOLA

Muhammad Aurang Zeb1, Abdul Halim

1*, Salim Ullah

1, Najeeb Ullah

1, Saadat Ullah Khan

1, M.Salahuddin

2, Mahjabin Rashid

3

1Department of Biochemistry, Hazara University Mansehra, Khyber Pakhtunkhwa, Pakistan

2Faculty of Medicine, University of Hong Kong, Pokfulam, Hong Kong

3College 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.

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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: [email protected]

Research Article (Open access)

International Journal of Life-sciences Scientific Research (IJLSSR), VOLUME 1, ISSUE 1


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



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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Health & Medicine

ANTIBACTERIAL ACTIVITY OF SKIMMIA LAUREOLA