International Journal of Life-Sciences Scientific Research (IJLSSR), VOLUME 1, ISSUE 1, pp: 33-36
SEPTEMBER-2015
Research Article (Open access)
ANTIBACTERIAL ACTIVITY OF SKIMMIA LAUREOLA 1
Muhammad Aurang Zeb , 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 pathogenic strains of bacteria. The hot and cold water extract of Skimmia laureola were used against four bacterial strains Escherichia coli,Bacillus subtilus, 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 laureola 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-
Out of which 20,000 plants species are believed to be used medicinally
tains substance that can be used for therapeutic purpose or which is a
in the third world (Mukherjee 2004). Approximately 6000 species of
precursor for synthesis of useful drugs.”(Sofowora, 1982).This definition
flowering plants occur in Pakistan and 700 of them have medicinal value
of medicinal plant has been formulated by WHO (World Health Organi-
(Stewart 1972). With an estimation of WHO that as many as 80% of
zation). The plants that possess therapeutic properties or exert beneficial
world’s population living in rural areas rely on herbal traditional medi-
pharmacological effects on the animal body are generally designated as
cines as their primary health care, the study on properties and uses of
“Medicinal Plants”. It has now been established that the plants which
medicinal plants are getting growing interests. In recent years this inter-
naturally synthesis and accumulate some secondary metabolites, like
est to evaluate plants possessing antibacterial activity for various diseas-
alkaloids, glycosides, tannins, volatiles oils and contain minerals and
es is growing (Clark and Hufford 1993). The screening of plant extracts
vitamins, possess medicinal properties. Medicinal plants are very impor-
and plant products for antimicrobial activity has shown that higher plants
tant for the cure of different microbial infections (Pennanen et al. 1996).
represent a potential source of new anti-infective agents (Smet 1997,
Plants have been utilized as medicines for thousands of years. These
Cowan 1999,). The leaves, flowers, fruits and roots are extensively used
medicines initially took the form of crude drugs such as tinctures, teas,
for treating cold, cough, whooping-cough (Dhuley 1999). Plant extracts
poultices, powders, and others herbal formulations. There are 4,22,127
have great potential as antimicrobial compounds, especially in the treat-
plant species growing on planet earth about 35,000 to 70,000 plants spe-
ment of infectious diseases caused by resistant micro-organisms (Nasir
cies are used as medicinal plants (Hasan et al. 2007).
and Chanda 2006). It is important to mention that over 75% of popula-
*
tion in Pakistan is cured by using traditional medicines prescribed by
Address for Correspondence:
more than 50,000 traditional herb practitioners.
Abdul Halim
The different system of eastern medicines that is, Unani, Ayurvedic and
Department of Biochemistry
homeopathy etc are entirely based on medicinal properties of these
Hazara University Mansehra
plants. The practice of traditional medicine is widespread in China, In-
Khyber Pakhtunkhwa, Pakistan.
dia, Japan, Pakistan, Sri Lanka and Thailand (Hasan et al. 2007). Today
Email:
[email protected]
herbal products and extracts are widely used to control various human Received: 03 August 2015/Revised: 14 August 2015/Accepted: 26 August 2015
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-
http://ijlssr.com IJLSSR © 2015 All rights are reserved
International Journal of Life-sciences Scientific Research (IJLSSR), VOLUME 1, ISSUE 1 tracts both gram positive as well as gram negative bacteria.
sterilize by means of autoclaving under the 15psi and 121 degree centi-
MATERIALS AND METHODS
grade temperature for 15 minutes in autoclave. After this media was
Collection of sample
poured aseptically into Petri dishes in laminar flow cabinet.
Skimmia laureola was obtained from Pansar Store near Lari Adaa, Man-
Preparation of plant extract
sehra (Figure 1). The plant was identified by the Department of Botany
For the attainment of maximum plant antimicrobial activity agar well
and Herbarium, Hazara University Mansehra, Khyber Pakhtunkhwa,
diffusion method was used. The plant was dried at sunlight and then
Pakistan.
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 Figure 1: Skimmia laureola
Selection of media
10 grams powder of Skimmia laureola plant was soaked in cold 100 ml
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.
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 centrifuged 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-
a) Composition of nutrient and simple agar Table: 1 Composition of nutrient agar Serial No. 1 2 3 4 5 6
a) Cold water extraction
carded the pure extract were then ready for antimicrobial sensitivity test. (Figure 3, 4 & 5).
Components Yeast extract Beef extract Peptones Glucose monohydrates Agar Sodium chloride
Table: 2 Composition of simple agar Electric shaker Serial No 1 2
Components
Centrifuge 5702
Cold water extraction of Skimmia laureola
Agarose Agaropectin
Figure: 3, 4 & 5 Electric shaker and Centrifuge &Cold water extraction of Skimmia laureola
Preparation of media
b) Hot water extraction
20 gram of nutrient agar was dissolved in 1 liter of distilled water in a
10 gram of Skimmia laureola powder was soaked in 100 ml distilled
conical flask and 4 grams of simple agar is also added and plugged in
water in a conical flask and then it placed in the incubator at 37degree
flask and shackedto mix well. Then it is heated on the hot plate stirrer to
centigrade for 12 hours after this it was placed in the hot water bath for 2
dissolve the media completely. The media and all glass ware swabs were
hours and then centrifuge it for 7 minutes at 4400 rpm then filtered it
http://ijlssr.com IJLSSR © 2015 All rights are reserved
International Journal of Life-sciences Scientific Research (IJLSSR), VOLUME 1, ISSUE 1 through filter paper and repeated this process for 3 times the supernatant obtained from 3
rd
Table 3: Cold water extraction of Skimmia leureola
time centrifugation collected while pellet were dis-
S.No
Micro-
Zone of inhibition in mm
Mean
Organism
carded and this was considered as a pure hot extract of these plants and these were ready for sensitivity tests. (Figure 6, 7 & 8 ).
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 development of new chemotherapeutic agents. The first step towards this goal is the in vitro antibacterial activity assay (Tona et al. 1998). Many reports Electric shaker
Centrifuge 5702
Hot water extraction of Skimmia laureola Figure: 6, 7 & 8 Electric shaker and Centrifuge & Hot water extraction of Skimmia laureola
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
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 antibacterial activity was checked. The zone of inhibition was measured by scale in mm after 24 hours the antibacterial activity were assigned according to the zone of inhibition produced by the plant extracts.
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 negative 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 respectively but did not show any antibacterial activity against Escherichia coli. The hot water extract of Skimmia laureola showed the antibacterial activity 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
RESULTS AND DISCUSSION
mirabilis.
Results
REFERENCE
In the present research project the antibacterial activity of Skimmia lau-
[1]
reola, were checked against four bacterial strains among which were Gram positive and were Gram negative. These strains included Staphylo-
Sofowara, A. 1982. Medicinal plants and traditional medicine in Africa. Wiley (Chichester, West Sussex, New York).
[2]
Pennanen TA, Frostegard, Fritz H (1996). Phospholipid fatty acid composi-
coccus aureus, E.coli, Proteus mirabilis and Bacillus subtilis. The hot
tion and heavy metal tolerance of soil microbial communities along two
and cold water extract of Skimmia laureola, were checked against the
heavy metal-polluted gradients in coniferous forests. Applied and Environ-
four bacterial strains. The inhibitory activity which were shown by cold
mental Microbiology, 62: 420-428. [3]
and hot water extract against the micro-organisms.
of Pakistan. Taxonomic Chemical methods. 01: 1-5.
Table: 2 Hot water extraction of Skimmia leureola. [4] S.No
Micro-
Zone of inhibition in mm
Hasan A, Khan MA, Ahmad M (2007). Authenticity of folk medicinal plants
Mean
Trivedi PC and Sharma NK. Etnomed. Palnt, pp. 18-33. African Journal of
Organism 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
Mukherjee TK (2004). Protection of Indian traditional knowledge. Editors,
Biotechnology Vol. 8 (24), pp. 7082-7086, 15 December, (2009) Available online at http://www.academicjournals.org/AJB ISSN 1684–5315 © 2009. [5]
Stewart RR (1972). History and exploration of plants of Pakistan and adjoining area, in: Flora of Pakistan. Nasir E, Ali SI National Herbarium, Pa-
http://ijlssr.com IJLSSR © 2015 All rights are reserved
International Journal of Life-sciences Scientific Research (IJLSSR), VOLUME 1, ISSUE 1 kistan Agricultural Research Council, Islamabad. pp. 69-75 & 89-123. [6]
Clark AM, Hufford CD (1993) Discco and development of novel prototype antibiotics for opportunistic infections related to the acquired immunodeficiency Kathmandu University Journal of Science, Engineering and Technology vol. 5 No. 1 January, 2009, pp 143-150. 150 syndrome In: human medical agents from plants american chemical society (ACS Symposium series 534) Washington D C pp 228-241
[7]
De Smet PAGM (1997). The role of plant-derived drugs and herbal medicines in Healthcare. Drugs, 54: 801-840.
[8]
Cowan MM (1999). Plant products as antimicrobial agents. Clin. Microbiol. Rev. 12: 564-582.
[9]
Dhuley JN (1999). Antitussive effect of Adhatoda vasica extract on mechanical or chemical stimulation-induced coughing in animals. J. Ethnopharmacol. 67: 361-65.
[10] Nasir R, Chanda S (2006). Activity of some medicinal plants against certain bacterial pathogenic strains, Phytochemical, Saurashtra University, Rajkot360005, Gujrat, Indian. [11] Srinivasan K, Natarajan D, Dheen MAN, Perumal G, Mohanasundari C, Prabakar K, Sengottuvel R (2006). Antibacterial activity of selected medicinal plants. Hamdard Medics XLIX. 2: 5-8. [12] Tona L, Kambu K, Ngimbi N, Cimanga K and Vlietinck AJ (1998). Antiamoebic and phytochemical screening of some Congolese medicinal plants. J. Ethnopharmacol., 61: 57-65.
http://ijlssr.com IJLSSR © 2015 All rights are reserved
