ABSTRACT: This work was done in order to investigate the microbial content of some bolted soft drivels sold in Enugu interlopes. Soft drink are consumed in large quantities by the different age groups, adult and children of different class of people all over the state. This is aided by the general availability of these drink almost every place roleve people gather for work or recreational activities. It is the appropriate to subject these soft drinks to some kind of investigation by isolation and characterizing the microorganism found. Simple Grows staining and biochemical test method were employed to isolate and identify the organisms found different bacterial and feast were identified these organisms include Fvlavobacterium spp. Bacillus Eschericjhism coli, Enteribacter arccosines Micro coccusroseus Micrococcus various Bacillus subtitles Aciuetobacter iwoffi, Micrococcus inteus Bacillus polymea, flavobacterium, rigeuse sacchenomyces, conevisciae klurenomyce sff, Torulopsis spp, cadida valida Geotrichuin candidium, sacchanoyces, carlbergersis cryptococus, albidus, Hausennlsa spp etc.
TABLE OF CONTENT
CHAPTER ONE
Introduction
CHAPTER TWO
Literature review
History and overview of soft drink industries
Production of soft drinks
Microbiology of soft drinks
Microbial growth
Factors affecting microbial growth
Ingredients and their effects
CHAPTER THREE
Material and method
Sample collection
Sterilization of glass warier.
Preparation of dividend and media
3.4.2 Preparation of serial / isolation
Identification and characterization of isolate organism
Colonial morphology
Cellular morphology
Grannies straining
Biochemical characteristic
Oxidase test
Citrate utilization test
Starch hydrolysis test
Nitrate reduction
Starch fermentation
CHAPTER FOUR
4.1 Result and discussion
CHAPTER FIVE
Conclusion and recommendation
Appendix
References
LIST OF TABLE
Table 1: colony counting
Table 2: cultural characteristic of bacterial growth on nutrient agar
Table 3: cultural characteristic of yeast growth on Pdp agar.
Table 4: result obtaining from gram staining and cellular morphology of bacterial isolates.
Table 5: result obtained from gram staining and cellular morphology of yeast isolates.
Table 6: different bacterial identified from the result of biochemical test (Appendix A& B)
Table 7: organisms (yeast) identified from the result of biochemical test. (appendix C & D).
CHAPTER ONE
INTRODUCTION
Soft drinks were defined by Harry and Wilbert (1984) as a class of non alcoholic beverages which may be divided into two classes carbondrate soft drinks some time referred to as Soda ;’soda pop “etc) and still or non- carbondrate soft drinks. soft drinks in general contains 86 to 92% water, 7 to 10% nutrient sweetbriers, carbodioxide (if present) acid and flavoring. The food and drug administration standard of identity for carbonated soft drink define them as a class of beverage made by absorbing carbodiocide in potable water. The amount of carbonhdioxide used is not less than that which well be absorbed by the beverage at a pressure of one atmosphere and a temperature of 60F. these drinks contain no alcohol, or only such alcohol (not in excess of 0.5% by weight of the finished beverage) as is contributed by the flavoring ingredient that may be use in carbonated soft drinks in such properties are reasonable required to accomplish their intruded effects. These include nutritive sweeteners flavoring ingredient natural and antificial colour additive acidifying agent buffering agent emulsifying stabilizing or viscosity producing agent foaming agent caffeine and chemical preservatives the beginning of the carbonated beverage (soft) drinks) as recorded by Resrosier (1977) date back to the closing years of the eighteen (18th ) century
Morris (1959) defined micro organisms as plant or animal organisms of microscopic or submicroscopic dimensions. Plant microorganisms are generally classified into three major groups, namely fungi bacteria and Algae as for as beverage manfacture is concerned the fungi are the most important group of spoilage organisms because yeast and mould belong to this group. The protozoa are the most important of the animal microorganisms.
Actually individual microorganisms are microscopic in size they can form colonies clouds precipitates sediments and scums with time and these are visible to the naked eyes and are the direct signs of the presence of microorganisms.
The microbiological safety and ratability of the diverse type of soft drinks products depend on their formulation (including) the use of chemical preservative carbonation low PH values and pasteurization) this was stated by ganglion (1982). Batchelor (1985) put it that the inter related environmental factors that determine the microbial load of product or the susceptibility of a product to microbial spoilage are intrinsic and Extrinsic factors.
Intrinsic factor are derived from the product and its formulation according to pauezai comments they are not easy to change Extrinsic factor are variable introduced during processing packaging distribution and storage. They should be easier to change and control. Girighano further recorded that control should be by application of the air Hazard Analysis critical Control point (HACCP) system to raw material formulating processing condition and packaging examples of such control would be :
Measurement of temperature during heat treatment of pasteurized product
Measurement of carbonation level by routine monvtorning of carbendioxide pressures in carbonated products
Measurement of pH water activity product viscosity and preservatives in combination with other formulation parameters
Maintenance of good hygienic practice to prevent the building up of spoilage organisms on the plant equipment and in the production areas.
Most microbiological problem arise because of poor quality of raw material (eg fruit concentrates sugar and syrup) poor processing hygiene packaging material and storage condition and these lead to over coming of the preservation system applied drinking manufactures or production of the beverages (soft drinks) by the micro organisms.
JUSTIFICATION
Soft drink are consumed in large quantities by the massed both male and female cutting across different age group adult and children of different classes of people all oven the state. This is aided by the general availability of these drinks almost every place where people gather for work or recreational activities so it is necessary to let the people taking this soft drinks know the consequences of leaving their drink open there by allowing some microorganisms which grow in the presence of air (acropea or acrophile organisms) to gain their way into the drink. Also for then to know the implicated organism in the soft drink thereby helping them to reduce their soft drink intake.
AIM AND OBJECTIVES
Aim and objective of this project is to investigation the microbial content of bottled soft drinks by:
Isolating the microorganisms present in them
Identify and characterize the microorganisms present
HYPOTHESES:
H0 Yeast and would are the only organisms implicated and identified in the microbiological charateristion of the three brand of soft drinks.
H1 Yeast and mould are the only organisms implicated and identified in the microbiological characterization of the three brand of soft drink, bacteria were also implicated and identified.
LIMITATION
This study was limited to Enugu metropolis and three leading and most consumed soft drink this was due to economic and transportation difficulties encountered in the course of the study.
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