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 species, Staphylococcus aureus, Lactobacillus plantarum, Micrococcus species, Lactobacillus brevis, Enterobacter species and Proteus species while the fungal isolates included Aspergillus species, Penicillium 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 × 107 cfu/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 ×105 cfu/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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