Antimicrobial activity of Cymbopogon Species against Human Pathogens

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ABOUT AUTHOR
Rudra Sharan Dwivedi *
Bhopal University,
Working in RMS Regrow, Pune
* rudsd1987@gmail.com

ABSTRACT
Cymbopogon
, is a tall perennial grass with 55 species ofgrasses. Lemongrass is the source of Lemongrass oil, natural citral, which is used as a basic raw material for synthesis of β-ionone used for synthesis of a number of useful aromatic compounds and Vitamin-A. In the present study the determined plant parts (5g) of four CymbopogonSps. were ground, extracted with five solvents and filtered after 48hrs. The dry weight of the extracts was obtained by allowing the solvent to evaporate and was used to determine concentration in mg/ml. Microbial susceptibility assays using the agar dilution (Mueller-Hinton Agar) method (%v/v and corresponding mg/mL values) and the Minimal Inhibitory Concentration (MIC) were carried out for E.coli, Bacillus sp., Streptococcus sp., Staphylococcus sp.

REFERENCE ID: PHARMATUTOR-ART-1726

INTRODUCTION
The plants that possess therapeutic properties or exert beneficial pharmacological effects on the animal body are generally designated as “Medicinal Plants” (theepicentre.com). Medicinal plants are gifts to nature to cure limitless number of diseases among human beings. The abundance of plants on the earth’s surfaces has led to an increasing interest in the investigation of different extracts obtained from traditional plant as potential sources of new antimicrobial agents (Bonjor et al., 2004). Natural antimicrobials can be derived from plants, animal tissues, or microorganisms (Gordon et al., 2001).  Medicinal plants and their parts contains different chemical substances that can be used for therapeutic purpose or which are precursors for synthesis of useful drugs. 

Medicinal plants synthesize substances that are useful to the maintenance of health in humans and other animals. These include aromatic substances, most of which are phenols or their oxygen-substituted derivatives such as tannins. Many are secondary metabolites, of which at least 12,000 have been isolated a number estimated to be less than 10% of the total (ayurvedicmedicinalplants.com). Secondary metabolites are organic compounds that are not directly involved in the normal growth, development or reproduction of organisms. Plants synthesize a bewildering variety of phytochemicals such as alkaloids, phenolics, terpenoids, glycosides etc.Lemon grass oil contains geraniol, myrcene, citronellal, limonene, linalool and dipentene and none of the constituents were reported to be mutagenic with Salmonella assay. (Gomes et al., 1998)

Cymbopogon, a tall perennial grass has 55 species. Cymbopogon is the source of Lemongrass oil, a good source of natural citral, which is used as a basic raw material for synthesis of β-ionone used for synthesis of a number of useful aromatic compounds and Vitamin-A. Lemon grass is a long thick grass with leaves at the top and a solid portion several inches long at the root end. Lemon grass used for culinary purposes, drugs, flavoring, insect repellants, liquors and perfumes. The lemon grass oil is used in the manufacture of perfumes of soaps, hair oils, scents and medicines. It also have antibacterial properties. Ionone prepared from the Citral present in lemon grass oil is one of the most important raw material for the preparation of vitamin-A(indg.in).An antimicrobial is a substance that kills or inhibits the growth of microbes such as bacteria, fungi, or viruses. 

There are several methods used to extract plant secondary metabolites (example: antimicrobial compound) which includes distillation, steam distillation, hydro distillation, water and  steam distillation, solvent extraction, carbon dioxide extraction, cold pressing, florasols/phytols, maceration. Solvent extraction involves separating the constituents of a mixture using a volatile solvent ( ethanolor hexane ) that does not mix with water. The solvent takes up the molecules to be extracted because it has a strong affinity with them. To collect the active ingredients, the solvent is then removed by distillation ( Panna et al;2008)

In this article, I give an account of the antimicrobial activity of 4 differerent species of Cymbopogon (Cymbopogon citratus, Cymbopogon flexuosus, Cymbopogon martini and Cymbopogon winterianus)

Sample collection:  Medicinal plants such as Cymbopogon citratus, Cymbopogon flexuosus, Cymbopogon martini and Cymbopogon winterianus were collected from different nursery of Bhopal.

Solvent extraction: 5gm of plant leaves were taken and dried under shade. Dry weights of the leaves were weighed and crushed using mortar and pestle. The crushed plant leaves were extracted with 25ml of different solvents like ethanol, methanol, propanol, acetone and diethyl ether. The crushed plant material kept in rotator shaker along with solvents for 2 days. After incubation, solvents were filtered using Whatmann No.1 filter paper. The residue after filtration was dried and weighed.

Collection of microorganisms:   The strains used in this work were isolated from patients suffered with bacterial diseases. The isolates were identified by Gram staining, biochemical characterization and by using selective media. The organisms isolated were E.coli, Bacillus sp., Staphylococcus sp. and Streptococcus sp. The microorganisms were grown overnight at 370C   in selective media such as, E.coli  - EMB agar, Bacillus sp. – Nutrient agar,  Staphylococcus sp.– Mannitol salt agar,Streptococcus sp. – Blood agar.

For identification and screening Gram’s staining was done. The morphologically identified organisms were grown on selective media such as, EMB agar, Starch agar, Mannitol salt agar and Blood agar .These selectively grown isolates were used for biochemical characterization by indole production, methyl red, Voges-Proskauer, citrate, SIM agar, gelatin, catalase, oxidase, Urease, nitrate tests.

Determination of antimicrobial activity:  Antimicrobial activity was determined by agar diffusion method. Three to five identical colonies of E.coli, Bacillus sp., Staphylococcus sp., and Streptococcus sp., from each agar plate were lifted with a sterile loop and transferred into a tube containing 5ml of nutrient broth. All the bacteria mentioned above were incubated at 37° C for 24hours. The Mueller Hinton agar was sterilized and cooled to 45-50° C and then the fresh cultures of the microorganisms (0.1ml) were inoculated. Each inoculated petridishes were swirled to distribute the medium homogeneously and allowed to dry for 15-20minutes. Wells (6) of 7mm were made into previously seeded MH agar plates. Each well was filled with 50µl of each plant extract. The same quantity of 75% ethanol served as control. The plates were allowed to stand at room temperature for 1hour for extract to diffuse into the agar and then they were incubated at 37° C for 24hours. Subsequently, the plates were examined for bacterial growth inhibition. The diameter of cleared zones was measured in cm. the resulting zone diameter is shown in the table. The transparently cleared zones containing micro colonies showed bacteriostatic activity.

Determination of minimal inhibitory concentration
This test was performed on the bacteria that exhibited inhibition zones using the effective plant extracts. The extracts that exhibited considerable activity when used in different concentration like 10µl, 20µl, 30µl, 40µl. The ethanol, methanol, propanol, acetone and diethyl ether extracts are used for the assay of MIC against E.coli, Bacillus sp., Staphylococcus sp., and Streptococcus sp. The test culture was inoculated with MH agar and poured in petriplates. The different concentration of each extract was poured in the wells. Then the plates were incubated at 37°C for 24hours. The lowest concentration that did not permit any visible growth, it was considered as the minimum inhibitory concentration.  

Medicinal Use
Cymbopogon
 is used extensively in Ayurvedic Medicine. It is normally used for common cold with headache, diarrhea, wind-cold impediment pain, cold pain in stomach duct and abdomen, knocks and falls.

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Websites
*ayurvedicmedicinalplants.com.
*mapbd.com.
*wikipedia.org
*motherherbs.com.
*theepicentre.com

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