Friday, 21 October 2016

A Research Proposal On Antimicrobial Activities Of Essential Oils On Some Food Spoilage Bacteria.

MADONNA UNIVERSITY,
ELELE, RIVERS STATE,
NIGERIA.
ANTIMICROBIAL ACTIVITIES OF ESSENTIAL OILS ON SOME FOOD SPOILAGE BACTERIA.

 BY

ODIRIH ONYINYE .E.
MB/12/031.
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. WESLEY BRAIDE
 MAY, 2016



CONTENTS

INTRODUCTION
AIM
OBJECTIVES
METHODOLOGY
STATISTICAL ANALYSIS
  
INTRODUCTION
According to World Health Organization (2007), consumption of foods contaminated with pathogenic microorganisms is a threat to human health. Essential oils are aromatic and volatile liquids extracted from plant material, such as flowers, roots, bark, seeds, peel, fruits, woods, and whole plant (Sanchez et al., 2010). Essential oils have been used for centuries in medicine, in food industry as flavours and in the cosmetic industry as fragrances, hand sanitizers, and almost about 3000 different essential oils are known, and 300 are used commercially in san In spite of the advances in the sanitation techniques and inspection services, the contamination of the foods with undesirable microorganisms is a potential risk during further processes, storage and distribution. Many of these oils have been shown to exert broad spectrum antimicrobial activity and even in laboratory tests, to kill or inhibit many foods – borne bacteria and this property have been an added benefit for their continued use Delamare et al., (2007). The use of chemical preservatives in the prevention of pathogenic and spoilage microorganisms in foods has lead to negative health effects. Microorganism have also be known to have acquired resistance against most of the chemical preservatives over the years, so the aim is to isolate these organisms and also asses their antibacterial effect of these essential oils with a view to establishing the possible role and enhancing food safety. 
AIM
The aim of the present study is to determine the antimicrobial activities of essential oils on some food spoilage bacteria.
OBJECTIVES
·        To study the antimicrobial effect of  essential oils on some food spoilage bacteria
·        To isolate, characterize and identify food spoilage bacteria from meat
·        To determine the phytochemical in plants
METHODOLOGY
      Collection of essential oil     
A total of 10 samples of commercial essential oil shall be obtained from Anthony Van Leeuwenhoek Research Center, Ihiagwa, Imo State.
        Isolation of Test Organisms
Pure cultures of different test organisms shall be obtained from meat samples. Serial ten folds dilutions shall be made up to 106 and approximately, 0.1ml aliquots of dilutions will be surface plated in duplicates on suitable medium. The plates shall be incubated at 37C for 24 h. Following incubation, the plates shall be examined and colonies characterized using standard methods.
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), Eosine methylene blue agar (EMB), Salmonella-shigella agar (SSA), nutrient agar (NA), MacConkey agar (MCA), and Deoxychocolate agar  (DCA)
 Obtaining Discs
A whatman No 1 filter paper shall be perforated to obtain a disc of 6mm diameter. The disc shall be sterilized in a hot air oven before use. Different concentrations of the essential oil diluted in DMSO shall be impregnated on the discs and allowed to dry.
Antimicrobial Activity    
The testing of the bacterial cultures for the inhibitory effect of essential oil shall be at different concentrations using dimethyl sulphur oxide (DMSO) as diluent. It shall be performed by using the disc diffusion method. The active cell suspension of the test organism standardized with equivalent of 0.5 McFarland shall be spread uniformly with the help of a sterile swab stick on Mueller-Hinton agar. Each experiment shall be done in triplicates and zone of inhibition measured in milliliter diameter shall be recorded and standard deviation calculated. Negative control shall be done using sterile distilled water in place of the essential oil and positive control using Ciprofloxacin. Plates shall be incubated at 37C for 24 hours before being examined for zone of inhibition. 

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