ABSTRACT
Effect of different carbon sources on the growth of antimicrobial producing Bacillus species in ogiri-egusi sold in Ogbeta, Orie, Artisian, Kenyatta, Enugu State were evaluated using standard microbiological and analytical methods. 12 bacteria were isolated and identified such as Bacillus subtilis,(33.33), Bacillus pumulis (41.66), Bacillus licheniformis (25) were identified based on their colony biochemical characteristics.The isolates of the highest occurrences is bacillus pumilus. Agar well diffusion method was used to determine the antimicrobial activity against two pathogenic organisms, Escherichia coli, Staphylococcus aureus. Bacillus subtilis and Bacillus pumulis, gave a zone of inhibition Escherichia coli 0.6mm and 0.8mm after 24 hours of incubation also carbon sources of 5% and 10% sucrose, Glucose and soluble starch were used to check their different carbon sources on the isolates using the spectrophotometer.5% had the highest optimum growth of 1.400 for glucose 1.545 for sucrose,2.376 for soluble starch which can be added in maximum quantity to a medium to produce antibiotics.
TABLE OF CONTENTS
Title page – – – – – – – i
Certification – – – – – ii
Dedication – – – – – – – iii
Acknowledgement – – – – – iv
Abstract – – – – – – – – vi
Table of contents – – – – – – – vii
List of tables – – – – – – – x
List of figures – – – – – – xi
CHAPTER ONE: INTRODUCTION
1.1 Aims and Objectives – – – – 1
CHAPTER TWO: LITERATURE REVIEW
2.1 Traditional Fermented Foods – – – – 8
2.2 Ogiri – – – – – – – 12
2.3 Detail of some Traditional methods Employed
Manufacture ogiri-egusi – – – – 13
2.4 Microbial Ecology – – – – – – 15
2.5 Microbiology of Ogiri-Egusi – – – – 17
2.6 Biotechnological Changes – – – – 18
2.7 Antimcrobial Activity on Bacillus – – – 20
2.8 Biotechnology of Ogiri – – – – 25
2.8.1 Effect of different carbon – – – – 26
CHAPTER THREE: MATERIALS AND METHODS
3.1 Materials – – – – – – – 27
3.2 Sample collection – – – – – 27
3.3 Preparation of samples – – – – – 27
3.3.1 Isolation Techniques – – – – 29
3.3.2 Streak plate methods – – – – – 29
3.4 Identification of isolates – – – – 29
3.5 Gram staining – – – – – – 29
3.5.1 Starch hydrolysis – – – – – 30
3.5.2 Oxidase enzyme activity – – – – 30
3.5.3 Voges – proskeur Test – – – – – 31
3.5.4 Citrate utilization Test – – – – 32
3.5.5 Sugar fermentation Test – – – – – 33
3.5.7 Methyl red – – – – – – – 34
3.6 Effect of Different Carbon sources – – – 35
CHAPTER FOUR
Results – – – – – – – – 36
CHAPTER FIVE: Discussion, conclusion, Recommendation.
5.0 Discussion – – – – – – – 45
5.1 Conclusion – – – – – — 47
5.2 Recommendation – – – – – 48
References – – – – – – – 49
Appendix I – – – – – – 61
LIST OF TABLES
Table one: Some traditional fermented foods………………………9
Table Two: Characterization and identification of the isolates……29
Table three: Table One: Biochemical properties of ogiri-egusi .. 38
Table four: Percentage occurrence of microorganisms…………….40
Table five: Antimicrobial activity of isolates…………………… .41
Table six: Effect of carbon source on isolates. …………………44
LIST OF FIQURES:
Figure one: Antimicrobial activity of Bacillus subtilis against Escherichia coli 0.6mm zone of inhibition……………………………………………………………………………..42
Figure two:Antimicrobial activity of Bacillus pumilu against Escherichia coli 0.8 zone of inhibition……………………………………………………………………..43
CHAPTER ONE
1.0 INTRODUCTION
Traditional fermented condiments (OGIRI-EGUSI) based on vegetable proteins are consumed by different ethnic groups in Nigeria, have been the pride of culinary traditions for centuries. It is evident that these products have played a major role in the food habits of communities in the rural regions serving not only as nutritious non-meat proteins substitute but also as condiment and flavoring agents in soup.
Traditional methods of manufacture should take advantage of biotechnological progress to assure reasonable quality and at the same time assure safety of these products. The requirements for a sustainable biotechnological development of Nigerian condiments are discussed in the scope of the microbiology and biochemical changes of the raw materials. Fermented vegetables, proteins have potential food uses as protein supplements and as functional ingredients in the fabricated food (Achi. 2005).
Seeds of legumes may account for up to 80% of dietary protein and maybe the only source of protein for some groups. Their cooked forms are eaten as meals and are commonly used as fermented form as meals and are commonly used in fermented forms as condiments to enhance the flavors of food (Odunfa, 1985). With high content of protein, legume condiments can serve as a tasty condiment to sauce and soups and can substitute for the food flavoring condiments are prepared by traditional methods of uncontrolled solid subtract fermentation resulting in extensive hydrolysis of the protein and carbohydrate components (Fetuga et al. 1973).
Fermented foods are essential parts of the world, particularly African (Odunfa. 1985). Fermentation is one of the oldest and most economical methods of producing and preserving foods in developed countries (David and Aderibigbe 2010). In Africa, many proteineous oily seeds such as cotton seed (Gossypium hirsutum), African locust bean (Parkia) and melon seed (Citrillus vulgaris) are fermented to produce soup condiments (Odunfa, 1981 ), which give pleasant aroma to soups and sauces. In many countries especially Nigeria and India
where protein calories. Malnutrition is a major problem, these condiments serve as food source of energy, low cost protein and fatty acids in diets (Odumodu. 2007).Ogiri is an oily paste produced by fermented melon seeds (Citrillus vulgaris) in the western part of Nigeria. Oyenuga (1986) have the composition of melon seed.
A melon seed has high protein and low Carbohydrate content. Citrullus vulgaris is a member of the family Cucurbitaceae (Alfred, 1986). Ogiri is characterized with very strong pungent odour. Among the consumers, there are preferences fir Ogiri produced from specific locality. The production process being a local art makes the quality varies. The fermented products are also stored at ambient temperature (28+2) oC. For varied length of time (days or weeks). The population and types of micro organisms involved during fermentation and storage could have affected the quality of the product.
Fermented foods are essential parts of diets in all parts of the world particularly Africa (Odunfa, 1985). Fruits, vegetables, cereals, root crops, legumes and oil seeds are used in the production of fermented food. Fermentation is one of the oldest and most economical methods
of producing and preserving foods in developing countries (David and Aderibigbe, 2010). In Africa, many proteinaceous oily seeds such as cotton seeds (Gossypium hirsutum), castor bean (Parkia biblobosa) and melon seed (Citrullus vulgaris) are fermented to produce food condiments (Odunfa, 1981 ), which gave pleasant aromas to soups and sauces. In many countries especially Nigeria and India where protein/calories malnutrition is a major problem, these condiments serve as good source of energy, low cost protein and fatty acids in diets (Odumodu, 2007). Thereby, supplement the nutritive quality of the respective diets where they consumed Ogiri is one of the condiments consumed in the Eastern and Western parts of Nigeria especially by the Ibos. Ogiri is an oily paste produced by fermenting melon seeds (Citrullus vulgaris) in the Eastern and Western parts of Nigeria. Oyenuga (1988) have the composition of melon seed to be dry weight (88.9%); crude protein (32.6%); ether extract (50.2%); crude fibre (3.7%); silica free ash (3.45%). Minerals (mg\100g) content of shelled melon seed were Calcium(112); Phosphorus (1777); Magnesium (578); Potassium
(538); Sodium (5); Chlorine (32); Vitamins (N/g); A (30.65); D (11.20) and E (0.25). Melon seed has high protein and low Carbohydrate content. Citrullus vulgaris is a member of the family cucurbitatea (Alfred, 1986).
Ogiri is characterized with very strong pungent odour. Among the consumer, there are preferences for Ogiri produced from specific locality. The production process being a local art makes the quality of the product varies. The fermented products are also stored at ambient temperature (28.2+2)oC for varied length of time (days or weeks),(David and Aderibigbe, 2010). The population and types of micro organism, involved in fermentation can result in food poisoning. Others are responsible for producing antibiotics (Obeta, 1983).
1.1 AIM AND OBJECTIVE OF THE STUDY
1) Isolation of antimicrobial producing Bacillus species in Citrullus vulgaris
2) To elucidate reasons data for preferences in ‘Ogiri’ from the four different market in Enugu town.
3) To identify the characteristic ability of microorganisms responsible for fermentation of Citrillus vulgaris to produce Ogiri
4) To identify the potential microorganisms and to study the effect of different carbon sources on isolates.
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