ISOLATION AND IDENTIFICATION OF MICROORGANISMS ASSOCIATED WITH SPOILT BAMBARA GROUNDNUT PUDDING ‘‘OKPA”

ABSTRACT

The project titled "Isolation and Identification of Microorganisms Associated with Spoilt Bambara Groundnut Pudding ‘Okpa’" aims to isolate and identify microorganisms present in spoiled Okpa, assess the potential health risks of consuming contaminated Okpa, and measure the microbial loads in spoiled samples. A total of 40 Okpa samples were collected from hawkers and markets in Umudike/Umuahia, wrapped aseptically, and analyzed in the laboratory. The study employed media such as nutrient agar, MacConkey agar, Salmonella Shigella agar, and Sabouraud dextrose agar for microbial culture. Sterility tests were conducted to ensure the media were contamination-free before inoculation. The samples were processed using the International Commission on Microbiological Specification of Foods (ICMSF, 2000) methods, involving serial dilution and spread plate techniques for bacteria and fungi cultures. The microbial load was calculated using the Total Viable Count (TVC) method. Various biochemical tests, including Gram staining, catalase, coagulase, oxidase, citrate utilization, indole, and sugar fermentation tests, were performed to identify the isolated microorganisms.

The results revealed several bacteria and fungi in the spoilt Okpa, including Staphylococcus aureus, Escherichia coli, Proteus species, Salmonella species, Shigella species, Streptococcus species, Bacillus species, Lactobacillus species, Pseudomonas species, Aspergillus, yeasts, Penicillium, Rhizopus, and Mucor. These microorganisms indicate potential contamination sources such as poor hygiene, environmental factors, and inadequate food handling practices. The study highlights the health risks associated with consuming contaminated Okpa, including foodborne illnesses caused by pathogens like Staphylococcus aureus and Escherichia coli.

The research emphasizes the need for improved hygiene and sanitary practices in the preparation and handling of Okpa to prevent contamination and ensure food safety. Recommendations include thorough cooking, using clean wrapping materials and utensils, maintaining a clean environment, and serving Okpa hot to reduce microbial loads. Additionally, educating food handlers on food safety and implementing strict monitoring by food inspectors are essential to prevent the sale of contaminated Okpa in markets and school environments. The study underscores the importance of these measures in preventing foodborne diseases and safeguarding public health.

TABLE OF CONTENTS

CHAPTER ONE

1.0       INTRODUCTION

1.1           Aims And Objectives

CHAPTER TWO

2.0       LITERATURE REVIEW

2.1       Description Of Bambara Groundnut

2.2       Origin And Distribution

2.3       Cultivation

2.4       Varieties

2.5       Production

2.5.1    Propagation And Planting

2.5.2    Growth And Development

2.5.3    Nutritional Requirement

2.5.4    Yield

2.5.5    Pest And Diseases

2.6       Uses

2.7       Nutritional Value

2.8       Processing

2.9        Bambara Groundnut-Effect Of Packaging

2.10        Microbial Spoilage Of Okpa

CHAPTER THREE

3.0       MATERIALS AND METHODS

3.1       Materials

3.2       Methods

3.2.1    Collection Of Materials (Okpa)

3.3       Sample Preparation/Sampling

3.4       Media Preparation

3.5       Sterility Test

 3.6      Culture Of Microbes From Okpa Samples

3.7       Identification Of Isolate

3.8       Gram Staining

3.9       Motility Test

3.10     Spore Staining

3.11     Biochemical Test

3.11.1  Catalase Test

3.11.2  Coagulase Test

3.11.3  Oxidase Test

3.11.4  Citrate Utilization Test

3.11.5  Indole Test

3.11.6  Sugar Fermentation Test

CHAPTER FOUR

RESULTS

CHAPTER FIVE

DISCUSSION AND CONCLUSION

5.1       Discussion

5.2       Conclusion

5.3       Recommendation

References

APPENDIX 1: Bambara groundnut plants (Vigna subterranean)

APPENDIX 2: Bambara groundnut plants (Vigna subterranean)

APPENDIX 3: Bambara groundnut plant (Vigna subterranean)

APPENDIX 4: Packaged Bambara Groundnut Pudding

APPENDIX 5: Steamed Bambara groundnut pudding

APPENDIX 6: Culture plates of microbes isolated from Bambara groundnut pudding “Okpa”

LIST OF TABLES

Table 1:           Morphology and Biochemical identification of Bacterial isolates

Table 2:           Identification of fungal isolates

Table 3:           Mean total aerobic bacterial count of okpa

Table 4:           Mean fungal count of Okpa

Table 5:           Occurrence of Bacteria Isolates in Spoilt Okpa Samples

Table 6:           Occurrence of Fungi Isolates in spoilt Okpa

LIST OF FIGURE

Fig. 1: Processes of Serial Dilution                                       

CHAPTER ONE

1.0       INTRODUCTION

Any food eaten has a direct influence on health. It is therefore an important task for food inspectors, food manufactures and food handlers to keep food safe from pathogenic microorganisms, especially when such foods are to be consumed without further processing that is ‘ready to eat’ foods.

A general observation of our society shows a social pattern characterized by increased mobility due to urbanization, large number of iterant workers and less family or home centred activities resulting in large percentage of the population depending on ready-to-eat foods for employment and food.

This situation however has resulted that food sanitary measures and proper food handling have been transferred from individual families to food vendors who rarely enforce such practices (Musa et al., 2002; Draper, 1996).

Okpa is a pudding/bean cake product of Vigna subterranean (L) verd which also is a ready to eat food after being steamed and it is eaten all over Nigeria and beyond (Lasekan and Akintola, 1987). Okpa is rich in phyto-proteins, vitamins and other nutrients and it is perfect meal with the right nutrient with good filling effect. Indeed, it makes delicious combination with most meals; it can be eaten with sell of rice or alone (Darlo et al., 1976).

Okpa preparation is basically the same all over Africa. It is made from Bambara groundnut flour which is usually made industrially by grinding the nut. But to make the paste simply soak the nuts, peel the skin and grind until it is smooth. Then pour the Okpa flour into a bowl, add the palm oil and mix thoroughly, until the flour changes from white to yellow. After which you add warm water, mix thoroughly until there is no lump, place it in a container of choice or banana leaves then leave for 45 minutes until it is done.

Furthermore, safe food storage temperate is difficult to maintain, since foods are often displayed over long periods and may not be reheated before serving. In other cases, sectors buy raw material from dubious source which may either or ready be contaminated with food borne pathogens or be unfit for consumption due to other reasons. However, microbial studies on ready to eat foods in American, Asian and African countries here revealed high microbial counts and a high incidence of food borne pathogens in food.

In Senegal, over 200 cases of food poisoning were traced from food made from dairy products (Rowland et al., 2000). Okpa being a ready to eat food and which can be contaminated in like manner has to be examined to determine its microbial quality.

            1.2           AIMS AND OBJECTIVES

The aims of this study are:

i)               To isolate and identify the microorganisms associated in ready to eat Okpa when contaminated or spoilt.

ii)             Possible health implications of eating contaminated or spoilt Okpa (Bambara groundnut pudding).

iii)           To check the microbial loads of the spoilt Okpa.

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