ISOLATION AND IDENTIFICATION OF LACTIC ACID BACTERIAL FROM KUNUN AYA (A FERMENTED NIGERIAN DRINK)

ABSTRACT

Samples of locally prepared Tiger- nut Milk (‘Kunun aya’) a fermented Nigerian drink was evaluated in this study The samples were separately subjected to proximate analysis, pH determination and Aerobic Mesophilic Bacterial Count using standard techniques to ascertain and compare their nutrient content, pH and microbial load respectively. Results of the proximate analysis as determined using standard procedures revealed that the locally prepared tiger nut milk (Kunun aya) had  moisture:71%, fat 1.63% crude protein 7.85% crude fibre; 0.33% ash, 0.35% and Carbohydrate 18.69%. The pH range of the samples were between 3.4 – 3.5while aerobic mesophilic bacterial count ranged between 5.4 x 106 to 1.086cfu/ml. Poor hygienic practice, poor quality tiger-nuts and water used in the preparation might be the cause of high microbial loads recorded. The microorganisms isolated were Bacillus specie Lactobacillus plantarum Staphylococcus aureus and Enterobacter species.

TABLE OF CONTENTS

Title page                                                                                                                    i

Certification                                                                                                                ii

Dedication                                                                                                                  iii

Acknowledgements                                                                                                    iv

Table of Contents                                                                                                       v

List of Tables                                                                                                              vii

Abstract                                                                                                                      viii

CHAPTER ONE

1.0              INTRODUCTION                                                                                        1

1.1       Aim                                                                                                                 3

1.2       Objectives                                                                                                       3

CHAPTER TWO 

2.0              LITERATURE REVIEW                                                                           4

2.2       Nutritional composition of tigernuts and its products                                                7

2.3       Tiger nut milk (kunun aya)                                                                              9

2.4       Flow chart of production                                                                                12

2.5       Microorganisms associated with tigernut milk (kunun aya)                           13

2.6       Lactic acid bacteria                                                                                         15

2.6.2    Contribution of lactic acid bacteria to food safety                                         17

2.6.3    Shelf life extension of lactic acid bacteria to fermented beverage                 19

2.6.4    Acid production                                                                                              19

2.6.5    Bacteriocins production                                                                                  21

2.6.6    Flavor formation                                                                                             22

CHAPTER THREE

3.0  MATERIALS AND METHODS                                                                      24

3.1       Isolation of lactic acid bacteria                                                                       24

3.2       Characterization and identification of isolates                                               24

3.3       Gram staining                                                                                                  25

3.4       Biochemical cultural characteristics                                                                25

3.4.0    Catalase test                                                                                                    25

3.5.1    Coagulase test                                                                                                 25

3.5.2    Citrate test                                                                                                      26

3.5.3    Motility, indole, urease test (MIU)                                                                 26

3.5.4    Triple sugar iron test                                                                                       27

3.5.5    Oxidase test                                                                                                    27

3.6       Sugar fermentation test                                                                                   27

3.7       Proximate analysis                                                                                          28

3.7.1    Moisture content determination                                                                      28

3.7.2    Total ash determination                                                                                  28

3.7.3    Crude protein determination                                                                           29

3.7.4    Fat content determination                                                                               29

3.7.5    Crude fibre determination                                                                              30

3.7.6    Carbohydrate determination                                                                           31

CHAPTER FOUR

4.0       RESULTS                                                                                                     32

CHAPTER FIVE

5.0       DISCUSSION, CONCLUSION AND RECOMMENDATION              37

5.1       Discussion                                                                                                       37

5.2       Conclusion                                                                                                      39

5.3       Recommendation                                                                                            40

References                                                                                                      41

Appendix

LIST OF TABLES

Table                                      Title                                                                            Page

4.1:      Total bacterial count                                                                                       33       

4.2:      The colonial morphology and biochemical characteristics of             34

the isolates

4.3:      pH values of the sample during formation                                                     36

4.4:      Proximate composition (%) of locally prepared kunun aya sample                36

CHAPTER ONE

1.0      INTRODUCTION 

Lactic acid bacteria (LAB) are groups of Gram positive, non-sporulating, anaerobic or facultative aerobic cocci or rods, which produce lactic acid as one of the main fermentation products of the metabolism of carbohydrates (Ocheme et al., 2011).

 Lactic acid bacteria (LAB) are naturally present in milk and milk products. LAB is generally associated with habitat rich in nutrients such as milk, cheese, meat, beverages and vegetables. In addition, (Adgidz, 2010) showed that lactic acid bacteria could be also isolated from soil, lakes, intestinal tract of animals and humans. Lactic acid bacteria (LAB) have been used for the fermentation of food and feed products since ancient days and today their major applications are still in the food and feed industry as starter cultures(Udeozor,2012).

Lactic acid bacteria are the most important bacteria in desirable food fermentations, being responsible for the fermentation of sour dough bread, fermented foods and beverages, all fermented milks and fermented vegetables. It plays an essential role in the production of all dairy products and is involved in the production of many other fermented foods and beverages, sausages, pickles, boza etc. Based on the end product of glucose fermentation lactic acid bacteria are grouped as either Homofermenters or Heterofermenters. The Homofermenters produce lactic acid as the major product of fermentation of glucose.(Oladele and Aina,2007)

LAB are recognized for their fermentative ability and thus enhancing food safety, improving organoleptic attributes, enriching nutrients and increasing health benefits. Many LAB species play an important role in the ripening process of cheese, especially to improve the consistency, aroma and flavor (Bosch and Alegna, 2005).

The origin of making milk from tiger nut is exclusive to the Spaniards to which it may have been introduced to by the Arabs. Tiger nut milk which is a beverage known as “Kunun Aya” in northern Nigeria is a refreshing healthy drink with many nutrients. It is a drink that gives energy and can be taken in any season of year, but more refreshing during the hot season. The Spanish call it chufa da harchata. Tiger nut milk is usually prepared raw and therefore is very rich in vitamin C and E and minerals such as phosphorus, magnesium, potassium, calcium, iron and also carbohydrates, unsaturated fats, protein and some enzymes, which aids digestion. This drink contains more iron, magnesium and carbohydrate than cow’s milk. It has advantage of not containing lactose, casein or sugar or protein in milk, or cholesterol and therefore ideal beverage for persons who do not tolerate gluten or cow’s milk (Odoemelan, 2003).

The search for lesser known and underutilized crops, many of which are potentially valuable as human and animal foods has been intensified to maintain a balance between population growth and agricultural productivity, particularly in the tropical and sub-tropical areas in the world (Odoemelan, 2003).

Tiger-nut was found to be a cosmopolitan perennial crop of the same genus as the papyrus plant.

Other names of the plant are Earth almond as well as Yellow-nut grass (Farhath et al., 2001; Belewu and Belewu, 2007).

1.1       Aim

The aim of this research work is to isolate and identify lactic acid bacteria from Tiger nut milk (Kunun aya)

1.2       Objectives

The objective of the study was to;

1.                  Isolation and identification of lactic acid bacteria

2.                  Evaluation of kunun aya nutritional quality.

3.                  Isolation and identification of mesophilic bacteria from kunun aya.

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