Monday, 23 October 2017

A RESEARCH PROPOSAL ON EFFECTS OF ESSENTIAL OILS ON MEDICAL PATHOGENIC BACTERIA

A RESEARCH PROPOSAL ON EFFECTS OF ESSENTIAL OILS ON MEDICAL PATHOGENIC BACTERIA
A RESEARCH PROPOSAL ON EFFECTS OF ESSENTIAL OILS ON MEDICAL PATHOGENIC BACTERIA

MADONNA UNIVERSITY,
ELELE, RIVERS STATE,
NIGERIA.

EFFECTS OF ESSENTIAL OILS ON MEDICAL PATHOGENIC BACTERIA

BY
O******* E*******
MB/12/***.

A RESEARCH PROPOSAL SUBMITTED TO THE
 DEPARTMENT OF MICROBIOLOGY, FACULTY OF SCIENCES, 
MADONNA UNIVERSITY IN PARTIAL FULFILMENT
 OF THE REQUIREMENTS FOR THE AWARD OF
 A BACHELOR OF SCIENCES (B.Sc.) IN MICROBIOLOGY

SUPERVISION: DR. W***** B*****
MAY, 2016.




CONTENTS
Introduction
Aim
Objectives
Methodology
References

  
INTRODUCTION
The increasing resistance of microorganisms to conventional chemicals and drugs has prompted scientists to search for novel sources of biocides with broad-spectrum activities (Abad, et al., 2007). In nature, Essential oil play an important role in the protection of plants. They may also attract some insects to promote the dispersion of pollens and seeds or keep away other undesirable insects. Thus, Essential oil can play a role in mediating the interactions of plants with the environment (Bakkali, et al., 2008).
Essential oils are aromatic liquid materials that are derived from plant material (flowers, buds, seeds, leaves, twigs, bark, stems, herbs, wood, fruits and roots) but are generally stored by the plant in secretory cells, cavities, canals, glandular trichomes or epidermic cells. (Van de Braak and Leijten, 1999). They can be obtained by expression, fermentation, effleurage or extraction with the method of steam being only for commercial purpose. Essential oils are concentrated natural products with strong smells that are produced by aromatic plants as secondary metabolites. These oils are present as variable mixtures of primarily terpenoids, especially monoterpenes (C10) and sesquiterpenes (C15), although diterpenes (C20) may also be present. Essential oils are liquid, volatile, limpid and coloured and are soluble in lipids and organic solvents that have a lower density than water. Essential oils are extracted from various aromatic plants that are generally found in temperate or warm countries, where they often represent an important part of the traditional pharmacopoeia. These plants may be known for their antioxidant effects as well as their antiseptic and medicinal properties and fragrance and are often used in the preservation of foods and as analgesics, sedatives, anti-inflammatories, spasmolytics and local anaesthetics (Bakkali, et al., 2008). Essential oils contain a wide series of secondary metabolites that can inhibit or slow the growth of bacteria, yeasts and moulds (De Martino et al., 2009). The Essential oils and their components have a variety of targets, particularly the membrane and cytoplasm, and in certain situations, they completely alter the morphology of the cells. Essential oils of medicinal plants have been used traditionally against pathogenic bacteria that caused infectious disease in human. In many parts of the world, the extracts and essential oil of medicinal plants with active biological compounds are used for their antimicrobial and antiviral properties that have been used in folk medicine (Hassawi and Kharma, 2006). The WHO considers phytotherapy in its health programs and suggests basic procedures for the validation of drugs in developing countries. Infectious diseases are the second leading cause of death worldwide (Fazly-Bazzaz et al., 2005). Generally, Gram-negative bacteria are more resistant to Essential oils than Gram-positive bacteria (Trombetta et al., 2005). The structure of the Gram-positive bacteria cell wall allows hydrophobic molecules to easily penetrate the cells and act on both the cell wall and within the cytoplasm. Phenolic compounds, which are also present in the Essential oil, generally show antimicrobial activity against Gram-positive bacteria.


AIM

This brief review will be to determine the effects of Essential oils against medical pathogenic bacteria
OBJECTIVES

  • To study the effect of essential oils on some medical pathogenic bacteria.
  • To characterize and identify medical pathogenic bacteria·
  • To determine the phytochemical properties in plants used for the extraction of essential oils.


METHODOLOGY

Collection Of Essential Oils
I will be using Essential oils which will be gotten from Anthony Van Leeuwenhoek Research Centre Ihiagwa, Imo State. Nigeria. 

Bacterial Strains
To evaluate the effects of the Essential oils, I will be using stocked isolates which will also be acquired from Anthony Van Leeuwenhoek Research Centre Ihiagwa, Imo State. Nigeria. They will be sub-cultured in the laboratory and prepared for antimicrobial susceptibility testing. Each test organism will be sub-cultured on different media for viability testing.


Characterization And Identification Of Bacterial Isolates
Pure cultures of bacterial isolates shall be identified on the basis of their colonial morphology, microscope and biochemical characteristics. Isolates shall be identified with reference standard bacteriological manual.

Media To Be Used


The media to be used in this study shall include Mannitol Salt Agar (MSA), Eosin Methylene Blue agar (EMB), Salmonella-Shigella Agar (SSA), Nutrient Agar (NA), MacConkey Agar (MCA), and Deoxychocolate Agar (DCA)


Media Preparations
I will prepare the media that will be used based on Manufacturers specifications. An electronic balance will be used to measure the quantity of agar that will be needed with the aid of a spatula before dispensing water into the beaker of choice which will autoclaved at 1210C for 15minutes before pouring into the number of petri dishes that will be used

Preparation Of Inoculums

I will be using streak method to inoculate my isolates on the media that will be prepared before incubating at room temperature (370C) for 24hours. After incubation, I will standardize my cultures using 0.5 McFarland standard turbidity of 1% Bacl and 1%H2SO4.

I will prepare Muller Hinton Agar using manufacturer’s specifications after autoclaving and then allow to solidify.

After that, I will use different swab sticks to pick from my inoculum, mix it in peptone water inside the bottle and then smear it evenly on of the Muller Hinton Agar plate and using a spreader, spread the inoculum evenly on the agar surface and then leave it to incubate at 370C for 24hours.

Preparation Of Antimicrobial Disc

I will impregnate sterile antimicrobial disc (Whatman 6mm) using Volume to Volume dilution to introduce the Essential oils to the disc. Ciproflaxacin will be used as my positive control disc and Dimethylsulphuric Acid (DMSO) as my negative control disc.

Disc Diffusion Method

I will place my disc that will be impregnated with Essential oils and standardized antibiotic disc on the surface of the standardized 0.5 McFarland inoculum and then wait for a period of 30minutes for absorption before incubating at 370C for 24hours and then check to see if there is/are susceptibilities or zones of inhibition.


REFERENCES

Abad, M.J., Ansuategui, M., Bermejo, P. (2007). Active Antifungal Substances from Natural Sources: ARCHIVOC 2007: 116–145.

Bakkali, F., Averbeck, S., Averbeck, D., Idaomar, M. (2008). Biological Effects of Essential Oils—a Review: Food Chem. Toxicol; 46: 446–475.

De Martino, L., De Feo, V., Nazzaro, F. (2009). Chemical Composition and In Vitro Antimicrobial and Mutagenic Activities of Seven Lamiaceae Essential Oils: Molecules; 14: 4213–4230.

Fazly-Bazzaz BS, Khajehkaramadin, M., Shokooheizadeh, HR. (2005). In Vitro Antibacterial Activity Of Rheum Ribes Extract Obtained From Various Plant Parts Against Clinical Isolates Of Gram-Negative Pathogens. Iranian J. Pharmacol. Res.; 2: 87-91.

Hassawi, D., Kharma, A. (2006). Antimicrobial Activity of Some Medicinal Plants against Candida Albicans. J. Biol. Sci.; 6: 109-114.

Trombetta, D., Castelli, F., Sarpietro, M.G., Venuti, V., Cristani, M., Daniele, C., Saija, A., Mazzanti, G., Bisignano, G. (2005). Mechanisms of Antibacterial Action of Three Monoterpenes. Antimicrob. Agents Chemother: 49; 2474–2478.

Van de Braak, S.A.A.J., Leijten, G.C.J.J., (1999). Essential Oils and Oleoresins: A Survey in the Netherlands and other Major Markets in the European Union. CBI, Centre for the Promotion of Imports from Developing Countries, Rotterdam: p. 116.


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