OCCURRENCE OF BACTERIA PATHOGEN IN TWO FRUIT JUICES AND THEIR CONTROL USING LACTIC ACID BACTERIA ISOLATE FROM UGBA

ABSTRACT

This project work studied the occurrence of bacteria in packaged fruit juices and their control with lactic acid bacteria isolated from fermented African oil bean seeds (pentaclethra microphylla) packaged fruit juices Ribena and Happy Hour were selected randomly from five market locations in Orieugba, Umuahia city centre, Ubani central market, Ahiaeke daily market and Umudike Mammy markets respectively and studied. Four bacteria species were isolated and identified from the fruit juices with varying levels of occurrences including Staphylococcus species (60%), Escherichia coli (12.5%), Bacillus species (40%) and Pseudomonas species (30%). Also, two lactic acid bacteria, Lactobacillus plantarum and Lactobacillus casei were isolated from randomly selected Ugba samples with occurrences of 100% and 66.7% respectively. Crude extracts from both LAB isolates demonstrated antibacterial activity against the packaged fruit isolates in which the inhibition zone of L. plantarum was in the range of 8.67mm (Bacillus species) to 14.67mm (E. coli) while that of L. casei ranged between 8.33mm (Bacillus) and 14.67mm (E. coli). The two test LAB inhibited Staphylococcus minimally with inhibition zone of 8.67mm and 10.33mm for the L. casei and L. plantarum respectively whereas their inhibition zone against Pseudomonas species were 11.67mm each for the LAB isolate. The overall antimicrobial activities of the LAB isolates were lower than the activity of standard antibiotic which caused inhibition in the range of 18.00mm (Pseudomonas species) to 24.67mm (E. coli). The relative potencies of the LAB isolate compared with the reference standard antibiotics were in the range of 46.43% (Bacillus species) to 64.83% (Pseudomonas species) for L. plantarum and 41.94% (Staphylococcus species) to 64.83% (Pseudomonas species) for L. casei. Both LAB isolates were deemed to have potentials for use in the control of the packaged juice bacteria isolates. 

TABLE OF CONTENTS

Title page                                                                                                                                i

Certification                                                                                                                            ii

Dedication                                                                                                                               iii

Acknowledgements                                                                                                                iv

Table of contents                                                                                                                    v

List of tables                                                                                                                           vii

List of figures                                                                                                                         viii

Abstract                                                                                                                                   ix

CHAPTER ONE

1.0       Introduction                                                                                                                1

1.1       Aim and Objectives                                                                                                    2

CHAPTER TWO

2.0       Literature Review                                                                                                       4

2.1       Fruit juice                                                                                                                   4

2.2       Health benefits of fruit juices                                                                                     4

2.3       Heat Resistant Fungi Spoilage of Pasteurized fruit juices                                         6

2.4       Preservation of fruit juices                                                                                         6

2.5       Factors affecting shelf life of juices                                                                           7

2.6       Deterioration of fruit juices by microorganisms                                                        7

2.7       Microorganisms Associated with Spoilage in Fruits and Juices                              8

2.7.1    E. coli (Escherichia coli)                                                                                            10

2.7.2    Salmonella species                                                                                                     10

2.7.3    Staphylococcus aureus                                                                                               11

2.7.4    Pseudomonas species                                                                                                 11

2.7.5    Serratia species                                                                                                          11

2.7.6    Yeasts                                                                                                                         12
2.7.7    Molds                                                                                                                          13

2.7.8    Bacteria                                                                                                                       14

2.7.9    Lactic Acid Bacteria                                                                                                   15

2.7.10  Acetic acid bacteria                                                                                                    16

2.7.11  Spore formers                                                                                                             16

2.8       Ugba and Microorganisms involved in the fermentation                                           16

2.9       Changes that occur during fermentation                                                                    19

2.10     Optimization of Ugba Fermentation                                                                          21

2.11     Microbiological safety of Ugba                                                                                  22

CHAPTER THREE

3.0       Materials and Methods                                                                                               24

3.1       Source of Material                                                                                                      24

3.2       Microbial Analysis of Fruit Juices (Screening for bacteria in fruit juices)             24

3.3       Media Preparation                                                                                                      25

3.4       Isolation of bacteria from fruit juices                                                                                     26

3.4.1    Characterization of Bacteria Isolates                                                                          24

3.4.2    Identification of Isolates                                                                                             29

3.5       Isolation of LAB from Ugba                                                                                      29

3.6       Antibacteria activity of LAB isolates against fruit juice bacteria isolates                        29

CHAPTER FOUR

4.0       Results                                                                                                                        30

CHAPTER FIVE

5.0       Discussion, Conclusion and Recommendations                                                        38

5.1       DISCUSSION                                                                                                             38

5.2       Conclusion                                                                                                                  42

5.3       Recommendations                                                                                                      42

References                                                                                                                  43

            Appendix                                                                                                                    48

LIST OF TABLE

Table 4.1:        Antibacterial activity of LAB isolates from Ugba against two fruit juice

bacteria isolate (mm)                                                                                      37

LIST OF FIGURES

Figure 3.1        Sketch of serial dilution of fruit juices                                                           24

Figure 4.1        Occurrence of bacteria in fruit juices sold in markets within Umuahia         34

Figure 4.2        Occurrence of LAB isolates in Ugba samples                                               35

Figure 4.3        Relative potency of LAB isolates from Ugba against bacteria isolates

of fruit juices                                                                                                  36

CHAPTER ONE

1.0    INTRODUCTION

Fruit juice refer to liquid extracted from edible fruits or fruit parts, preserved fruits or any concentrate of such liquids (Codex-Stan, 2005), commercial fruit juices are available in many different types of packaging usually designed to ensure the availability of the juices in the same farm (Abbslurt, 2005).  Available research information show that both natural and synthetic preservatives are sometimes added to fruit juices to enhance their quality and shelf life (Anand and Sati, 2013; Rowe et al., 2012). Microorganisms including coliforms find their ways into fruit juices during processing especially through water (Tagmin, et al., 2010). In the same cause of production, different methods are applied to preserve the juices including the chemicals, freezing, canning, pasteurization etc (Fasoyiro et al., 2005).

The existence of microorganisms in fruit juices are said to be responsible for their fermentation, spoilage and food borne diseases (Essien et al., 2014). According to Inhamakov et al.,(2013), microbial contamination  occur in fruit juices due to errors in packaging, and on application of aseptic principles during processing, standard production regulation demands commercial fruit juices to comply with correct processing, storage and constant surveillance procedures (Igbal et al., 2016). The report of Splistoeaser et al., (1994), the presence of microorganisms in fruit juices originates from different sources which include the natural flora of the raw material (fruits), handlers and those that enter by chance inoculation from the surrounding of the processing environment. Other researchers observed  that the reason for presence of microorganism in processed fruit juices is due to the fact that high temperature heat treatments have deliteriory  effect on organoleptic properties of juices hence processors tends to apply limited heat which does not  like microbial spores thereby leaving residual microorganisms emanating from the spores. But Teli et al., (2014) observed that some microorganisms in fruit juices do not have any deleterious effects hence the juices are consumed without harm whereas stews may manifest in spoilage, fermentative or food borne diseases.  

There are reports of food borne diseases associated with consumption of fruit juices in several places (Clumber et al., 2007; Gosh et al., 2007; Lewis et al., 2006; Mosupye and Holy, 2000). Also, available records show that illnesses attributed to bacteria like. Coli, Shigella, Salmonella, Staphylococcus aureus have been recorded from consumed fruit juices (Barro et al., 2006; Sandeep et al., 2001). Again many reports show the isolation, many bacteria species in fruit juices within the bonetry. Olurofuwon et al., (2011) isolated many genera of bacteria from local fruit juices sold in Sagamu Ogun State including  Websolla, E. coli, Shigella, Salmonella, and Pseudomonas,while Olamiyi (2013) isolated many bacteria species from NAFDAC approved packaged fruit juices in Ilorin, Kwara state.

Lactic acid bacteria (LAB) are widely applied in the food industry due to their safe level and their probiotic potentials. This is partly due to reported cases of side effects with most chemical preservation. Against the above information, the project was designed to study the incidence of bacteria in two commercial fruit juices sold within Umuahia and its environ and the antibacterial activity of LAB isolates from Ugba against them.

1.1       Aim and Objectives

The aim of this project is to study the occurrence of bacteria in two fruit juices sold in Umuahia area and the effect of crude extracts (supplements) LAB isolate from fermented African oil bean seeds on them.

Objectives

The objectives of this project work are to carry out the following:

1.     To isolate LAB from Ugba

2.     To isolate and identify bacteria from two commercial fruit juices sold in Umuahia and its environ.

3.     To determine the antibacterial activity of crude extracts from LAB isolates from Ugba against the fruit juice bacteria isolates.

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