ISOLATION AND IDENTIFICATION OF BENEFICIAL BACTERIAL SPECIES

ABSTRACT

This study focused on the isolation and identification of beneficial bacterial species, particularly Lactic Acid Bacteria (LAB), from various fermented food products including yoghurt, Akamu, and Ogiri. The primary aim was to discover alternative and cheaper sources of essential nutrients crucial to human nutrition, such as vitamins and proteins. The methodology involved preparing samples of these fermented foods, followed by serial dilution and culture on MRS agar. The bacterial isolates were then characterized through colony morphology, microscopic features, and a series of biochemical tests to ascertain their identity.

Nine bacterial isolates were obtained from the samples, with a focus on evaluating their nutrient-producing capabilities. The isolates were tested for their ability to synthesize essential vitamins such as Thiamine (Vitamin B1), Riboflavin (Vitamin B2), and Niacin (Vitamin B3). The results revealed that Riboflavin was the most abundantly produced vitamin, with a concentration of 10.04 mg/100g, followed by Niacin at 2.79 mg/100g, while Thiamine was produced in much smaller quantities (1.12 mg/100g). The production of proteins by these isolates was relatively low, with the highest yield being 0.01 mg/100g. Notably, none of the isolates demonstrated the ability to produce starch, which was consistent with prior findings in microbial studies.

The findings of this study align with previous research indicating that LAB, primarily isolated from fermented products, can serve as an alternative source of vital nutrients, particularly vitamins. The study concludes that LAB holds potential for use in nutritional supplementation, especially in resource-limited settings where conventional nutrient sources may be scarce or expensive. However, further research is needed to explore the industrial application of these bacteria in nutrient production and to enhance their protein synthesis capabilities

TABLE OF CONTENTS

CHAPTER ONE

INTRODUCTION

1.1     Background of the Study

1.2     Aims and Objective of the Work

CHAPTER TWO

LITERATURE REVIEW

2.0     What Bacterial Actually Mean

2.1     Lactic Acid Bacteria

2.2     Acetic Acid Bacteria: A Second Group of Bacterial Importance in Food Fermentati

2.3     Bacteria of Alkaline Fermentation

2.4     Bacteria and the Soil: Nitrogen Fixation:

2.5     Human and Bacteria

2.6     Bacteria and Food Industry

2.7     Bacteria in Other Technologies

2.8     Lactic Acid Bacteria

2.8.1 Taxonomical Classification of Lactic Acid Bacteria

2.8.2 General Position of Specific Lactic Acid Bacteria

2.8.3 Occurrence of Lactic Acid Bacteria in Nature:

2.8.4 Antimicrobial Properties of Lactic Acid Bacteria

2.9     Measurement Issues

2.9.1 Aerosol Sampling Issues   

2.9.2 The Problem of Culturability

2.9.3  Enumeration without Culture

2.9.4 Measurement of Bacterial Emission Fluxes

2.10   Mechanism of Emission to the Atmosphere

2.11   High Altitude Concentration Measurement

2.12   The Effect of Climatological Variable on Ambient Bacteria

2.12.1 Temperature

2.12.2 Relative Humidity

2.12.3 Wind Speed

2.12.4 Rain Events

2.12.5 Total Particulate Mass

2.12.6 Diurnal Cycle

2.13   Bacterial Aerosol Over Land by Ecosystem

2.13.1 Crops, Rural and Remote Continental Location

2.13.2 Forests

2.13.3 Desert

2.13.4 Grass Lands 

2.13.5 Shrub

2.13.6 Urban

2.13.7 Coastal.

2.14 Bacteria in Marine Air

2.15 Probiotics:

2.15.1 Safety

CHAPTER THREE

MATERIALS AND METHODS

3.1     List of Materials

3.2     Source of Materials

3.3     Medium Preparation

3.4     Sample Preparation

3.5     Isolation and Identification of Microbes   

3.6     Characteristics of Isolates

3.7     Colony Features/Morphology

3.8     Microscopic Features/Gram Stain Reaction of Isolates

3.9     Biochemical Reaction Tests

3.10   Sugar Utilization Tests

3.11   Identification of Isolates

3.12   Screening of the Isolates for the Production of Essential Nutrients

3.12.1  Determination of Vitamin B1

3.12.2 Determination of Riboflavin (Vitamine B2)

3.12.3 Determination of Niacin (Vit B3)

3.12.4 Determination of Starch

3.12.5 Determination of Protein

CHAPTER FOUR

RESULTS

4.1     Occurrence of Lactic Acid Bacteria in Fermented Foods (Akamu Yoghurt and Ogiri)

4.2     Nutrient Production by Lactic Acid Bacteria Isolated From Fermented Foods

CHAPTER FIVE

DISCUSSION AND CONCLUSION

5.1     Discussion

5.2     Conclusion

References

LIST OF TABLES

Table 1:          Showing occurrences of Lactic Acid Bacteria in fermented foods

Table 3:          Characterization of lactic acid bacterial isolates

Table 2:          Colonial Mophology of Isolates

Table 4:          Niacin production by Lactic Acid Bacteria isolated from fermented foods

Table 5:          Riboflavin production by Lactic Acid Bacteria isolated from fermented foods

Table 6:          Thiamine production by Lactic Acid Bacteria isolated from fermented foods

Table 7:          Showing protein production by Lactic Acid Bacteria isolated from fermented foods

CHAPTER ONE

INTRODUCTION

1.1 BACKGROUND OF THE STUDY

Lactic acid bacteria were first discovered by Scheele (1789) from sour milk. Pasteur discovered in 1857, that the souring of milk was caused by the microorganisms. Lactic acid was first produced commercially by M/s Clinton processing company, USA. Lactic acid bacteria have been widely used for the fermentation of many fermented product such as cheese, sourdough, buttermilk, brined vegetables, yoghurt and sauerkraut (Salim et al., 2006).

Lactic acid bacteria are widespread in nature and predominant microflora of milk and its products. Lactic acid bacteria are one of the important groups of microorganisms in food fermentation. A wide variety of strains are routinely used as starter cultures to manufacture dairy products such as curd, cheese, whey and yoghurt (Ayad et al., 2004). These bacteria produce organic acid hydrogen peroxide and several enzymes during fermentation (Fredrick and Luc, 2004). These compounds not only contribute to desirable effect on food flavor and texture, but also inhibit undesirable microflora and extending shelf life of products. Growth of spoilage and pathogenic bacteria in the fermented foods were inhibited due to the production of antimicrobial substances by lactic acid bacteria as their competition for nutrients

Beneficial bacteria are essential for good health- indeed, they are essential to life itself as bacterial are so intertwined with our body’s ability to function that you would actually die without them. Life is not aseptic. Beneficial bacterial interact with your immune system. In an interesting theory called the hygiene hypothesis, researchers have discovered that an overly sanitary home, birth and environment (i.e. lack of exposure of micro biota) may contribute to ante immune   diseases and healthy children eat dirt. In effect, beneficial bacterial actually train your benefit.

They also play critical role in your body’s ability to fully absorb the nutrient you eat. They break down potentially negative components of your diet like oxalic acid, manufacture natural vitamins and perform a slew of their duties that keep you well and healthy.However, the beneficial role of these bacterial is not restricted to within the body, it goes far beyond the internal body or even external body of animals and humans, as we all know that the body is in desperate need of major nutrients for its  survival and proper functioning. The nutrient required and consumed by man is immune even too numerous to mention, these beneficial bacterial acts on and degrade so many sources (substrates) to produce many essential nutrients required by man.

Nowadays, enormous amount of agriculture and industrial cellulosic wastes have been accumulating in environment. Celluloses are regarded as the most important renewable resources for bioconversion. Many cellulose substance were hydrolysed to simple sugars for making single cell protein, sweetness, etc. it have become the economic interest to develop am effective method  to hydrolyze the cellulosic biomass.

Cellulases are the inducible bioactive compounds produced by microorganism during their growth on cellulosic matters (Mukesh et al; 2012)

Beneficial bacterial are screened for the production of antibiotics because they (some of them) posses antimicrobial potential, hence they are screened and identified for the Novel production of antibiotics.

However, these bacterial are industrially beneficial  in the production of many other nutrients, too many to be mentioned here, from lots and lots of substrates, hence this project is mainly geared towards isolating, identifying and screening of these bacterial species, from various sources and using them to produce some important identifiable nutrients from various sources.

1.2    AIMS AND OBJECTIVE OF THE WORK

The aims of this project are geared towards:

      i.         Isolation and identification of beneficial bacterial species which are not harmful to man, from various sources.

     ii.         Screening these bacterial species for their capacity to produce these essential nutrients sources.

   iii.         It aims to discover alternative as well as cheaper sources of some essential nutrients which forms major part of human nutrients.

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