STUDIES ON MICROBIAL SUCCESSION DURING UGBA (FERMENTED SEEDS OF PENTACLETHRA MACROPHYLLA BENTH) PRODUCTION

ABSTRACT

This study was carried out to determine the succession of microorganisms in fermented african oil bean seed.   The samples were prepared by two traditional methods, packaged in sterilized plastic container and fermented at two different temperature (ambient 28±20C) and 37.50C. Proximate analysis , physicochemical analysis, protein solubility and microbial analysis were carried out. These analyses were carried out at 24 hourly interval for 96 hours. The samples were prepared by two traditional methods, packaged in sterilized plastic container and fermented at two different temperature (ambient 28±20C) and 37.50C them identification of isolate The result showed an increase in protein, fat, crude fibre and ash contents of the samples after 96 hours of fermentation with an incr ease in pH value and a decrease in carbohydrate content for all samples after 96 hours fermentation. The peroxide values, iodine values, and saponification values of the oils decreased while the protein solubility content increased with fermentation time. The result of the total viable counts were 2.74×1017 Cfu/g, 2.34×1017 Cfu/g, 2.18×1017 Cfu/g, 2.9×1017 Cfu/g, for sample BT1 (boiled twice and fermented at 28 ± 2 oC ambient temperature), BT2 (boiled twice and fermented at 37.5 oC), B1 (boiled once and fermented at 28 ± 2 oC ambient temperature), BO1 (boiled once and fermented at 37.5 oC), respectively after 96 hours fermentation. No mould growth was found in the unfermented and fermented Ugba .Microorganisms isolated were Proteus sp, Bacillus Subtilis, Klebsiella Sp Staphylococcus Aureus and other bacillus species . Organoleptic changes in texture and colour were seen to be a factor of period of fermentation and storage.Result indicates that ugba spoilage is primarily a result of continued activity of B.subtilis ,B.lichenformis and B.pumilis. the main objective is to study microbial succession in fermented oil bean seed(Ugba).

TABLE OF CONTENTS

 TITLE                                                       PAGE                                                         

 Title Page                                                                                                                 i

Approval Page                                                                                            ii

Certification                                                        iii

Dedication                                                                      iv

Acknowledgement                         v

Table Of Content                                                           vi

List Of Tables                                                                             x

List Of Figures                                                                                     xi

Abstract                                                                               xii

CHAPTER ONE: INTRODUCTION                                                                                                                       

1.1  introduction                                                                                                                        1

1.2 Statement of Problem                                                                                                                      3

1.3 Research Objectives                                                                                                                      4

HAPTER TWO: LITERATURE REVIEW                                                    

2.1 Importance and description of African Oil Bean Tree                                                                                                                      5

2.2 Preparation of Ugba                                                                                                                      7

2.3 Micro – Organisms involved During Fermentation                                                                     9

2.4 Chemical Composition of the Seeds                                                                                        10

2.5 Nutritional Value of Fermented and Unfermented African Oil Bean Seed (Ugba)                            13

2.5.1 Vitamin and Mineral Content                     13

2.10   Handling and Storage of African Oil Bean Seed                                                                     19

CHAPTER THREE: MATERIALS AND METHODS

3.1     Study Area                                                                                                                                21                                                                                                     

3.2      Preparation of Ugba                                                                                                  21

3.3      Proximate Analysis                                                                                                   24

3.3.1    Moisture Content Determination                                                                                      24

3.3.2    Ash Content Determination                                                                                             24

3.3.3    Crude Protein Determination                                                                                            25

3.3.4    Fat Content Determination                                                                                              26

3.3.5     Crude Fiber Content Determination                                                                                 27

3.3.6     Determination of Carbohydrate                                                                                         27

3.4       Physico- chemical Analysis                                                                                                 27

3.4.1 Iodine Value Determination                                                                                              27

3.4.2 Saponification Value Determination                                                                                    28

3.4.3 Peroxide Value Determination                                                                                         29

3.4.4 pH  Determination                                                                                                           30

3.4.5 Protein Solubility Determination                                                                                         30

3.5   Microbiological Analysis and  Biochemical Tests                                                                31

3.5.1 Total viable count                                                                                                                31

                                                                       vii

3.5.2 Mould count                                                                                                              31

3.5.3 Identification of Isolates                                                                                                        32

3.5.4 Gram Staining                                                                                                              32

3.5.5 Motility Test                                                                                                                 32

3.5.6 Oxidase Test                                                                                                                 33

3.5.7 Citrate Utilization Test                                                                                                            33

3.5.9 Indole Test and                                                                                                               34

3.5.9   Methyl Red Test                                                                                                           35                                                                                                                            

3.6       Sensory Evaluation                                                                                                   35

3.7       Data Analysis                                                                                                             36

CHAPTER FOUR: RESULT AND DISCUSSION

4.1       Changes in chemical composition of African oil bean seed (Ugba) during                                      37

fermentation                                                                                                                  

4.1.1    Moisture content                                                                                                      37

4.1.2     Ash                                                                                                                           38

4.1.3    Fat                                                                                                                           38

4.1.4    Protein                                                                                                                       39

4.1.5    Crude fibre                                                                                                                40

4.1.6    Carbohydrate                                                                                                             40

4.2.0    Physico- chemical properties                                                             41

                                                      LIST OF TABLES

Table                                                 Title                                   Page

Table 1: Uses of Pentaclethra macrophylla Benth in West African            7 

Table 2: Major Micro – Organism Encountered in Fermentation of Sliced Oil

bean Seed (per gram)                     10

Table 3: Amino Acid content (g/100g protein) of African Oil Bean Seed         12

Table 4: Fatty Acid Composition of African Oil Bean Seed            13

Table 5: Mineral and Vitamin Content of Unfermented and Fermented Ugba                            14

Table 6: The Medicinal Uses of African Oil Bean Seeds plants  (Pentaclethra macrophylla Benth)   19

Table 7: Changes in total viable count (TVC) of Ugba during fermentation                 46

Table 8: Total viable microbial count from four different fermented ugba sample                       47

Table 9: Identification and characterization of Microbial Isolates from fermented ugba Samples                    48

Table 10: Identification and characterization of Fungal Isolates        49

Table 11: Percentage occurrence of fungal and bacterial isolates from fermented ugba samples       50

Table 12: Proximate Composition of Fermenting African Oil bean Seeds (ugba)                            51

Table 13: Temperature, Ph and Titrable Acid Values of the sample during formation                                           52

Table 14: Sensory evaluation of the fermented African oil bean seed (Ugba)  54

                                           LIST OF FIGURES    

Figure                                          Title                                                   Page             

Fig 1: A Flow Diagram for the Preparation of Ugba                               23

CHAPTER ONE

INTRODUCTION

1.1 Introduction

Ugba is the Igbo name for sliced fermented African oil bean seed (Pentaclethramacrophylla  Benth).The African oil bean seed is called several names in Nigeria, such as “Apara” by the Yoruba, “Ugba” or ” Ukpaka” by the Igbo’s (Enujiugha and Akanbi, 2005). It is consumed mostly in the eastern states of Nigeria as a local delicacy popularly known as “African salad” prepared with oil, pepper, fish and salt and also prepared with tapioca, stockfish and garden eggs. It can be eaten with boiled or roasted yam and cocoyam (Okafor et al., 1991; Mbajunwa et al., 1998). Ugba is a traditional food generally prepared in homes as a small family business. Its method of preparation varies from one place to another resulting in a non-uniform product (Njoku and Okemadu, 1989). According to Enujiugha (2000), “Ugba” is produced traditionally by boiling the seeds overnight for easy removal of the seed coat. The cotyledons are sliced and cooked until they are soft with reduced bitter taste. The sliced “Ugba” is washed about 5 times or more and fermented for 3 days (Enujiugha, 2000).

According to Enujiugha (2003), the cooked, processed and fermented seed “Ugba” is used to prepare some delicious African soup and sausages for eating different staples. Enujiugha (2003) also noted that it is a rich source of vitamins and minerals, having a high demand for local and export consumption. Previous research showed that fermentation gives a better nutritional product than the raw seed (Achinewhu, 1986; Enujiugha and Olagundoye, 2001). According to Siggel and Faucet (1976), fermentation is the oldest method of processing legumes. Other workers discovered that less fermented foods are produced and consumed in Nigeria and only few studies have been carried out on them. Onwuliri et al. (1986) and Onwuliri and Ekepenyong (1994) reported the chemical composition of Pito and Burukutu a known Nigerian traditional brand of alcoholic beverage brewed from sorghum and millet, while another worker studied the fermentation of cassava. Enujiugha and Akanbi (2002) reported that fermentation of African oil bean seeds to produce “Ugba” softens the cotyledon, improves its digestibility and nutrients availability. Obeta (1983) showed that “Ugba” is a cheap and important source of protein especially for people whose staple foods are deficient of proteins. The consumption of the Ugba could pose as a means of addressing the prevailing protein energy malnutrition (PEM) in developing countries (Enujiugha and Akanbi, 2008). It is a cheap and available source of plant protein in developing countries of the world. Its protein contains 20 amino acids and 80 % of its fatty acids are also essential (Enujiugha and Agbede, 2000; Ikediobi, 1981). “Ugba” is a low acid food and a product of alkaline fermentation (Enujiugha and Akanbi, 2005). The rise in pH into the alkaline region has been attributed by Njoku and Okemadu (1989) to activities of various hydrolytic enzymes.

In many Eastern communities in Nigeria, “Ugba” is consumed as a meat analogue due to its high protein content, the locally prepared” Ugba” is done through a mixed wild bacteria fermentation of the sliced, boiled and soaked African Oil bean seeds. Its raw seeds are bitter and contain antinutritional factors like paucine, cyanide, oxalate, saponin, phytate and tannins (Achinewhu, 1983; Enujiugha and Akanbi, 2005; Onwuliri etal., 2004). Thermal treatment of the seeds increases the nutrient bioavailability and the digestability and drastically reduces the level of antinutritional compounds while increasing iron, calcium, potassium, thiamine and riboflavin levels (Enujiugha and Ayodele-oni, 2003). Obeta (1983) and Odunfa and Oyeyiola (1985) Observed that the source of micro-organisms in fermenting seeds comes from handling processing, utensils used in processing and the leaves used in packaging. The result of this uncontrolled fermentation of “ugba” is a very short shelf- life product that lasts for 3-4 days, thereby, limiting the availability of “Ugba”. This is due to the activities of micro-organisms involved in the fermenting process that remains viable till the product is consumed or continues fermentation till the product becomes unacceptable. The product deterioration or spoilage is evidenced by organoleptic changes in colour, texture, odour and taste (Mbata and Orji, 2008). In this research work, a detailed study will be carried out on improving nutritional quality, freshness and general acceptability of “Ugba”. The chemical composition of hygienically processed “Ugba” fermented at different temperatures will also be evaluated. The aim is to study microbial succession in fermented oil bean seed (Ugba)

 1.2   Aim and  Objectives

The aim of this  research   is to study microbial succession in fermented oil bean seed (ugba). The  objectives are:

   i.     To evaluate the proximate composition of Ugba during the fermentation period .

ii    To evaluate the physico-chemical characteristics of Ugba at different fermentation temperatures and days.

iii      To monitor the microbial population of Ugba fermented at different temperatures and days.

iv      To determine the general acceptability of Ugba produced under different     fermentation temperatures (sensory evaluation)

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