MICROBIOLOGICAL EVALUATION OF UGBA

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ABSTRACT

This study was aimed at evaluating the microbiological quality of ready-to-eat Ugba sold within Umuahia metropolis. A total of fifty (50) samples of Ugba were collected from different sales point comprising of twenty-five (25) nylon packaged Ugba and twenty-five (25) leaf wrapped Ugba in five (5) different markets within Umuahia metropolis. The samples were processed aseptically in sterile peptone water and diluted serially in the 10-fold serial dilution. Appropriate diluents were inoculated into Nutrient agar for total bacteria count. The samples were also inoculated into MacConkey agar for total coliform count, de Man Rogosa Sharpe agar for lactic acid bacteria count and Sabouraud dextrose agar for fungal count. The bacterial isolates identified based on their morphological and biochemical characteristics were Bacillus speciesStaphylococcus aureusLactobacillus plantarumMicrococcus speciesLactobacillus brevisEnterobacter species and Proteus species while the fungal isolates included Aspergillus speciesPenicillium species and Mucor. From the present study, the microbial count of Ugba revealed that the samples had a total heterotrophic bacteria count which ranged from 1.38 × 10cfu/g to 2.61 × 107 cfu/g. The total coliform count  ranged from 6.1×105 cfu/g to 1.16 ×106 cfu/g. The total lactic acid bacteria count  ranged from 5.8 × 105 cfu/g to 1.14 × 106 cfu/g. The total fungal count  ranged from 1.7 ×105 cfu/g to 3.8 ×10cfu/g. It was observed in this study that Bacillus species is the most frequently occurring isolate with a high percentage occurrence of 26(52.0%), followed by Staphylococcus aureus at 15(30.0%), Lactobacillus plantarum at 14(28.0%), Micrococcus species at  9(18.0%), Proteus species at 7(14.0%), Lactobacillus brevis at 6(12.0%) and also Enterobacter species at 4(8.0%). Among the fungal species, Mucor was the most predominant and occurring at 24(48.0%), followed by Penicillium species at 13(26.0%) and also Aspergillus species at 12(24.0%). Conclusively, this study revealed facts about Ugba showing its poor shelf life associated with uncontrolled fermentation and poor packaging on the products and also underscores the need to maintain improved hygiene standards and good manufacturing practices (GMP) in the preparation of Ugba to minimize the microbial contamination of Ugba.







TABLE OF CONTENTS

Title Page                                                                                                                                            i

Certification                                                                                                                                          ii

Dedication                                                                                                                                       iii

Acknowledgements                                                                                                                           iv

Table of Contents                                                                                                                               v

Lists of Figures                                                                                                                                   vii

Lists of Tables                                                                                                                                  viii 

Abstract                                                                                                                                              ix                                                                                         

CHAPTER ONE: INTRODUCTION

1.1 Background of study                                                                                                                    1

1.2 Objective of This Research                                                                                                                  2

1.2.1 Aim of This Research                                                                                                               3

 CHAPTER TWO: LITERATURE REVIEW

2.1 Nature of The Plant and The Seed                                                                                               4

2.2 Preparation Method of Ugba                                                                                                        5

2.3 Chemical Composition of The Seed                                                                                            8

2.4 Nutritional Values of The African Oil Bean Seed                                                                       8

2.5 Benefits of The African Oil Bean Seed (Ukpaka)                                                                       9

2.6 Toxicology of The African Oil Bean Seed                                                                              11

2.7 Changes Associated With Fermenting African Oil Bean Seed                                                  11

2.8 Microbiological Changes During Fermentation                                                                         13

 

CHAPTER THREE: MATERIALS AND METHODS

3.1 Study Area                                                                                                                                  16

3.2 Sources of Samples                                                                                                                     16

3.3 Sample Collection and Preparation                                                                                             16

3.4 Sterilization of Glasswares                                                                                                          17

3.5 Preparation of Culture Media                                                                                                    17

3.6 Microbiological Analysis of Samples                                                                                                       17

3.7 Isolation and Characterization of Bacteria                                                                                  18

3.7.1 Isolation of Microorganisms                                                                                                   18

3.7.2 Characterization of Isolates                                                                                                     18

3.7.3 Gram Staining                                                                                                                          19                                                                                                                        

3.8 Biochemical Test                                                                                                                       19

3.8.1 Catalase Test                                                                                                                           19

3.8.2 Coagulase Test                                                                                                                        19

3.8.3 Oxidase Test                                                                                                                           19

3.8.4 Indole Test                                                                                                                              20

3.8.5 Urease Test                                                                                                                              20

3.8.6 Citrate Test                                                                                                                              20

3.8.7 Hydrogen Sulphide Test                                                                                                         20

3.8.8 Endospore Staining Test                                                                                                         21

3.8.9 Starch Hydrolysis Test                                                                                                            21

3.8.10  Sugar or Carbohydrate Fermentation Test                                                                           21

3.9 Fungal Identification                                                                                                                  22

CHAPTER FOUR: RESULTS

Results                                                                                                                                              23

CHAPTER FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATION

5.1 Discussion                                                                                                                                    30

5.2 Conclusion                                                                                                                                  32

5.3 Recommendation                                                                                                                     33

REFERENCES








 

LIST OF FIGURES


FIGURE                   TITLE                                                     PAGE


1                       Flow Chart of the Preparation of Ugba                        7                                                                                 



 

 

 

LISTS OF TABLES


TABLE                                       TITLE                           PAGE            

1 Total Heterotrophic Microbial Counts of Ugba Samples from Five Different Markets              25

2 Colonial Description and Biochemical Characteristics of the  Isolates                                        26

3 Macroscopic and Microscopic Characteristics of Fungal Isolates                                                   27

4a Percentage Occurrence or Frequency of Isolation of the  Bacterial Isolates                              28

4b Percentage Occurrence or Frequency of Isolation of the Fungal Isolates                                  29

 

                                                              

 




CHAPTER ONE

INTRODUCTION


1.1 BACKGROUND OF STUDY

There are various plant seeds that are fermented and used as food in some rural and urban parts of Nigeria, among which is Ugba from African oil bean (Pentaclethra macrophylla). African oil bean seed also known as “Ugba” and “Ukpaka” in Igbo language is a popular food delicacy in Nigeria especially among Igbo ethnic group. It is an alkaline-fermented product rich in protein and is obtained by a solid-state fermentation of the boiled, shredded seed of African oil bean tree (Pentaclethra macrophylla benth). Ugba is an essential food item for various traditional delicacies where it is mixed with slices of boiled stock fish (Ugba and okporoko). Ugba as a source of protein in developing countries of the world and Africa in particular is of primary importance because it is cheap and available. Fermented seeds are not just palatable but serve as a delicacy amongst consuming regions where it is consumed and garnished with other vegetables or staples. Consumption of fermented Ugba seeds could bridge the prevailing protein energy malnutrition (PEM) marasmus in developing countries (Enujiugha, et al., 2010). African oil bean tree (Pentaclethra macrophylla benth) is a woody plant predominant in the rain forest areas of West and Central Africa belonging to the family; leguminosaea and sub-family; mimosoidae. The seeds are oval, flat and black to grey in color. The seeds are composed of 44-47% oil, 36.2-43.89% protein and 4-17% carbohydrates. Unprocessed seeds are bitter and possess anti-nutritional factors amongst which are paucine, cyanide, oxalates, saponin, phytic acid, phytate and tannins. Processing of these seeds involves boiling, removal from pod, shredding or cutting into slices, further boiling, sieving, wrapping in banana or plantain leaves and fermentation. Thermal treatment induces a resultant rise in nutrient bioavailability and seed digestibility. Processing Ugba seeds drastically reduces the levels of the anti-nutritional compounds mentioned while increasing iron, calcium, potassium, thiamine and riboflavin levels (Ike et al., 2016). Fermentation is a chemical change in food brought about by enzymes from living microorganisms. Fermented foods are prepared from plant and animal materials by processes in which microorganisms play important role in modifying the substrate physically, naturally and sensorily. Some desirable changes in foods (especially legume based) which occur during fermentation include changes in texture and organoleptic characteristics (flavors, aroma and appearance or consistency); especially elimination of beany flavors, improvement in digestibility, enhancement in the quality of the product, improved safety (absence of toxins and partial or complete elimination of nutritional factors), increased nutritional quality and reduced cooking time. Flavor components are the primary contributors to product acceptability either desirable or undesirable, if fermentation is uncontrolled (Kabuo et al., 2013). The fermentation of the African oil bean seed effects better nutrient availability and digestibility with significant softening of the cotyledons. With successive processing steps during the fermentation, there is progressive softening of the cotyledons, reduced astringency and increased palatability, and enhanced meaty flavor. However, the fermented product Ugba, has a high rate of deterioration and susceptibility to microbial spoilage within two weeks of production. The major concern has been to seek ways of lengthening the shelf life without lowering the nutritional quality, freshness and consumer acceptance of  Ugba (Enujiugha et al., 2010).


1.2 OBJECTIVE OF THIS RESEARCH

The Objective of this work is to study microbial diversity and to determine the predominant microorganisms involved in the fermentation of African oil bean in five different markets in Umuahia metropolis by enumeration of the organisms and isolation of the bacteria.


1.2.1      AIM OF THIS RESEARCH

The growth and occurrence of organisms of public health importance in Ugba is of great concern. Natural fermentation process used routinely in the fermentation process of Ugba allows participation of diverse microorganisms which may include contaminants. This study was therefore carried out to investigate:

1.     The microbial load of ready-to-eat Ugba offered for public health consumption in Umuahia and its potential to pose risk to public health.

2.     To isolate, characterize and identify microorganisms associated with the products Ugba in Umuahia.

 

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