ANTIMICROBIAL ACTIVITIES OF GRAPE FRUITS SEEDS (CITRUS PARADISI) EXTRACT USING STAPHYLOCOCCUS AUREUS AND ESCHERICHIA COLI AS TEST ORGANISM
    ABSTRACT
Antimicrobial activities of grape fruit seed extract ws investigated. The organisms used includes Staphylococcus aureus and Escherichia coli which belong to gram positive and gram negative organisms. Disc plate methods were used to determine the antibacterial activity of the grape fruit seed extract on the selected bacterial and gave a positive result. Analysis showed that there were lethal effects on the test organism after 48hours showing that the disc impregnated with the extract hail an inhibitory effect on the test organisms; Staphylococcus aureus and Escherichia coli which was indicated by a clear zone.
TABLE OF CONTENTS
CHAPTER ONE
1.0    Introduction    -    -    -    -    -    -    -    
1.1    Statement of problems -    -    -    -    -    -    
1.2    Aims of the study    -    -    -    -    -    -    
1.3    Significance of the study     -    -    -    -    -    
1.4    Scope of the study -    -    -    -    -    -    -    
1.5    Limitation -    -    -    -    -    -    -    -    
CHAPTER TWO
2.0    Literature review    -    -    -    -    -    -    
2.1    Historical background of grape -    -    -    -    
2.2    Components of grape fruit -    -    -    -    -    
2.3    Effective features of grape fruit -    -    -    -    
2.4    Important Application of grape fruit
2.5    Factors affecting antibacterial
CHAPTER THREE
3.0    Materials and method    -    -    -    -    -    
3.1    Reagents and apparatus-    -    -    -    -    -    
3.2    Collection of text organism    -    -    -    -    
3.3.0    Preparation of culture media -    -    -    -    
3.3.1    Plating technique (spread plate)    -    -    -    
3.3.2    Pour plate method-    -    -    -    -    -    
3.3.3    Microbial count    -    -    -    -    -    -    
3.3.4    Gram staining    -    -    -    -    -    -    
3.4    Preparation/extraction of the active ingredients -    
3.5    Determination of the extracts    -    -    -    -    
3.6.0    Phytochemical screening of the seed extract –    
3.6.1    Test for Tannins    -    -    -    -    -    -    
3.6.2    Test for Saponnies    -    -    -    -    -    
3.6.3    Test for Flavonoids     -    -    -    --    -    
3.6.4    Test for Cardiac Glycosides    -    -    -    -    
3.6.5    Test for Anthraquinone    -    -    -    -    
3.6.6    Test for Alkaloids    -    -    -    -    -    -    
3.7.0    Biochemical test -    -    -    -    -    -    
3.7.1    Catalase test     -    -    -    -    -    -    
3.7.2    Coagulate test    -    -    -    -    -    -    
3.7.3    Indole test    -    -    -    -    -    -    -    
3.7.4    Methyl red test    -    -    -    -    -    -    
3.7.5    Motility test    -    -    -    -    -    -    -    
3.7.6    Sugar fermentation    -    -    -    -    -    
CHAPTER FOUR
4.0    Results    -    -    -    -    -    -    -    -    
CHAPTER FIVE
5.1    Discussion     -    -    -    -    -    -    -    
5.2    Conclusion         -    -    -    -    -    -    
5.3    Recommendation -    -    -    -    -    -    -    
    References     -    -    -    -    -    -    -    
    Appendix     -    -    --    -    -    -    -    -    
LIST OF TABLES
Table 1:    Morphology characteristics and confirmatory test of bacteria isolated    -    -    -    -    
Table 2:     Biochemical test of the isolated bacteria -    
Table 3:    Effect of grape seed against staphylococcus aureus     -    -    -    -    -    -    
Table 4:     Effect of grape seed against Escherichia coli -    
Table 5:     Phytochemical test result     -    -    -
CHAPTER ONE
1.1    INTRODUCTION
Citrus paradise belongs to the Ructaceae family and is grown into two belts on both sides of the equator, from about 20 to 40 degrees latitude. It produces edible fruit called grape fruit.
All citrus is though to originate in the Himalayan region of south western china and Northern India. Columbus brought seed of citrus to the Western hemisphere in 1493 and planted them first on the island of Hispaniola, now called Haiti. Citrus became commercialized in America in the late 1805. Brazil has today become the dominant market leader of citrus (Morto, 2000). Citrus may also be processed into other fruit products such as fruit drinks, fruit beverages, nectar wine, cardinal, vinegar, canned fruits, syrup, pectin, jellies, James, fruit butters (Hagedorn, 2002).
The grape fruit is a subtropical citrus tree grown for its fruit which was originally named the forbidden fruit of Barbados. The fruit was first documented in 1750 by Rev. Griffith Hughes describing specimens from Barbados in the study, antimicrobial activities of the seed extracts of grapes were examined against Escherichia coli and Staphylococcus aureus, all tested bacteria were inhibited by the grape seed extracts using agar well diffusion method. The most sensitive  of the bacteria was A hydrophila while hasandede grape seed extract was the most inhibited extract on test bacteria. Hasandede seed extract at 10% concentration gave largest inhibition zone diameter. (Dr. Jacob, 2000). However, grape seed were powdered and the fatty. Material was extracted in a soxhelt extractor with petrlleum ether. The deflated powder was extracted with acetone.
Anecdotal claims for an antimicrobial effect are loosely supported by small abstract scientific experiment which apparently show some antimicrobial activity in vitro assays such as the agar diffusion. However, there is considerable scientific evidence that the antimicrobial activity associated with grape fruit seed extract is attributable to the contaminate or adulteration commercials grape seed extract preparations with synthetic antimicrobials.  After careful evaluation of the anecdotal claims for an anti-microbial effect, the preliminary studies state that the citricidal brand of grape seed extract was used for testing. Despite the anecdotal antimicrobial claims, the citricidal brand as well as the nutribiotic brand were both contaminated with synthetic benzethonium chloride that was implausible to be made from grape fruit seeds. Components of grape fruit juice is also referred to as the anti scurvy vitamin but lacks in most Africa diet alongside other vitamins and mineral (Melian, 2003).  
In such circumstance the vitamins in concentrated from is a therapeutic (Corinne, 2000). Grape extracts prevent potentials in cancer, cardiovascular disease and many other disorders.
1.2     STATEMENT OF THE PROBLEMS
•    Anti-microbial substances are agents that act antagonistically against Escherichia coli and Staphylococcus aureus.
•    It is hoped that many people will achieve better health by using grape fruit seed extract in prevention and treatment of diseases.
1.3    AIMS OF THE STUDY
•    To investigate into the microbial activity of grape fruit seed extract using Staphylococcus aureus and Escherichia coli.
1.4    SIGNIFICANCE OF THE STUDY
This work is indeed of colossal significance in many respects. These includes.
-    To provide technical and microbiological information for the grape fruit seed production in large scale.
-    To update the knowledge of unit of operation involved in the production of grape fruit.
-    To highlight to preventive potentials of grape fruit seeds.
-    To know the microbial activity of grape seed as an antimicrobial agents.
-    Alternative medication for bacterial infection.
1.5    SCOPE OF THE STUDY
This research work is confirmed to the antimicrobial activity of grape fruit using Escherichia coli and Staphylococcus aureus.
1.6    LIMITATION
This research work was limited to antimicrobial activities of gape fruit seed using Escherichia coli and staphylococcus as test organism. However, the extract was not tested on human injcted by the organism but it is believed that prior to its potency on the test organism, it would be able to cure human and animal diseases infected  by the organism. However the scope of this subject does not subject us to ascertain the minimum concentration of the organism extract that can inhibit the growth of the organism and thus was ignored