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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