MICROORGANISMS ASSOCIATED WITH HOMEMADE FRUIT JUICES

ABSTRACT

Microbiological analysis of some selected homemade drinks sold in Umudike was done. Four (4) popular homemade drinks, zobo, kunu, orange and pineapple drinks were used.10 ramdonly selected drinks of each type were collected in each of three market study area including Ahiaeke, Ndoro and Umudike and analyzed accordingly. Results show that the average bacteria load of the drinks were 1.78x105cfu/mL(zobo), 1.98x105cfu/ml(kunu), 1.82x105cfu/mL(orange drinks) and 1.63x105cfu/mL(pineapple). The ranges were 1.57x105cfu/ml to1.94x105cfu/ml in zobo, 1.73x105cfu/ml to 2.36x105cfu/mL for kunu, 1.73x105cfu/ml to 1.92x105cfu/ml for orange and 1.43x105cfu/ml for pineapple drink. The bacteria flora show deviate species with varying levels of occurrences including Lactobacillus (33.3), Salmonella (6.7), Staphylococcus (17.7), Pseudomonas (33.3), E. coli (16.7), Streptococcus (17.7) and Bacillus (46.7)for zobo. Lactobacillus (56.6), Salmonella (23.3), Staphylococcus (93.3) Pseudomonas (33.3), E. coli (30.0), Streptococcus (46.6) and Bacillus (53.3) for kunu. Lactobacillus (33.3), Salmonella (3.33), Staphylococcus (86.6) Pseudomonas (26.6), E. coli (3.30), Streptococcus (46.6) and Bacillus (30) for orange and Lactobacillus (23.3), Salmonella (0.00), Staphylococcus (66.6), Pseudomonas (30), E. coli (0.00), Streptococcus (0.67) and Bacillus (23.3) for pineapple .The fungi load was an average of 3.44 x103cfu/ml in zobo been in the range of 3.33x103cfu/ml to 3.67x102cfu/ml. The figure for kunu was in the range of 7.67x103cfu/ml to 1.01x104cfu/ml. Orange drinks were between 2.67x102cfu/ml and 3.33x102cfu/ml while pineapple drink had between 5.67x102cfu/ml and 8.33x102cfu/ml. The fungi isolate include yeast (55.83%), Aspergillus (23.32%), Penicillum (17.48%), and Rhizopus (14.99%).it was observed that the microbiological quality of the home made drinks sold in Umudike was poor as all had microbial load above the maximum permissible load reputation.

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 and Objectives                                                                                                    3

CHAPTER TWO

2.0       Literature Review                                                                                                       4

2.1       Microbiological Analysis of Some Packaged Fruit Juices                                         4

2.2       Microbial Quality of Commercially Packed Fruit Juices In South-East Nigeria    6

2.3       Microbiological Quality of Some Locally-Produced Fruit Juices In Ogun

State, South Western Nigeria                                                                                     7

2.4       Food-Borne Pathogens in Fruit Juices                                                                       9

2.4.1    Staphylococcus aureus                                                                                               10

2.4.1.1 General Characteristics and Taxonomy of Staphylococcus aureus                                    10

2.4.1.2 Source and Transmission of Staphylococcus aureus                                                  10

2.4.1.3 Diseases Caused by Staphylococcus aureus                                                               11

2.4.2    Bacillus cereus                                                                                                           11

2.4.2.1 General Characteristics and Taxonomy of Bacillus cereus                                       11

2.4.2.2 Habitat and Transmission of Bacillus cereus                                                             12

2.4.2.3 Diseases Caused by Bacillus cereus                                                                           12

2.4.3    Salmonella spp.                                                                                                           13

2.4.3.1 General Characteristics and Taxonomy of Salmonella spp                                        13

2.4.3.2 Habitat and Transmission of Salmonella spp.                                                                        14

2.4.3.3 Diseases Caused by Salmonella spp.                                                                          14

2.4.4    Shigella spp.                                                                                                               15

2.4.4.1 General Characteristics and Taxonomy of Shigella spp                                            15

2.4.4.2 Source and Transmission of Shigella spp.                                                                  16

2.4.4.3 Diseases Caused by Shigella spp.                                                                               16

2.4.5    Coliform Bacteria                                                                                                       16

2.4.5 Escherichia coli                                                                                                             17

2.4.5.1 General characteristics and taxonomy of Escherichia coli                                        17

2.4.5.2 Diseases Caused by Escherichia coli O157: H7                                                        18

CHAPTER THREE

3.0       Materials and Methods                                                                                               19

3.1       Collection of Samples                                                                                                19

3.2       Sterilization of Materials                                                                                            19

3.3       Preparation of Culture Media                                                                                     19

3.4       Inoculation and Isolation                                                                                            20

3.5       Purification of Isolates                                                                                               20

3.6       Identification of the Isolates                                                                                       20

3.7       Gram Staining                                                                                                            21

3.8       Biochemical Test                                                                                                        21

3.8.1    Catalase Test                                                                                                               21

3.8.2    Indole Test                                                                                                                  21

3.8.3    Citrate Utilization Test                                                                                               22

3.8.4    Hydrogen Sulphide (H₂S) Production Test                                                                22

3.8.5    Starch Hydrolysis                                                                                                       22

3.8.6    Motility, Indole, Urease (MIU)                                                                                  23

3.8.7    Coagulase Test                                                                                                           23

3.8.8    Oxidase Test                                                                                                               24

3.10     Identification of Fungal Isolates                                                                                 24

3.10.1  Colonial Morphology                                                                                                 24

3.10.2  Yeast Viability Staining                                                                                             24

CHAPTER FOUR

4.0       Result                                                                                                                          25

CHAPTER FIVE

5.0       Discussion, Conclusion and Recommendation                                                          33

5.1       Discussion                                                                                                                   33

5.2       Conclusion                                                                                                                  35

5.3       Recommendations                                                                                                      35

References                                                                                                                  36

LIST OF TABLES

CHAPTER ONE

1.0       INTRODUCTION

Juices are the aqueous liquids expressed or otherwise extracted usually from one or more fruits or vegetables, purees of the edible portion of one or more fruits or vegetables, or any concentrates of such liquids or purees (Fraternale, 2011). Fruit juices are beverages and nutritious drinks with great taste and which are commonly consumed for their refreshing attributes, nutritive values or vitamin content and health benefits (Nwachukwu and Aniedu, 2013; Rashed et al., 2013). According to Tasnim et al. (2010), fruit juices are becoming an important part of the modern diet in many communities. They are nutritious beverages and can play a significant part in a healthy diet because they offer good taste and a variety of nutrients found naturally in fruits. For example, orange juice is rich in vitamin C which is an excellent source of bio-available antioxidant phytochemicals as reported by Franke et al. (2005) and significantly improves blood lipid profiles in people affected by hyper-cholesterolemia (Kurowska et al., 2000; Tasnim et al., 2010). Fruit juices promote detoxification in the human body. Juices are fat-free, nutrient-dense beverages rich in vitamins, minerals and naturally occurring phytonutrients that contribute to good health (Tasnim et al., 2010).

Fruit juices are available in their natural concentrations or in processed forms. Juice is prepared by mechanically squeezing fresh fruits or is extracted by water. A whole fruit can be directly squeezed, macerated or crushed so as to produce a considerable amount of pulp or juice or may be extracted by water. The extracted juice could be used in their natural state or could be concentrated by evaporation or freezing and could be preserved by chemical means, bottling, canning or freezing (Oranusi et al., 2012). The constituents of processed juices are mainly water, sugar, preservatives, colour, and fruit pulp. The most commonly used preservatives are benzoic acid, sorbic acid or sulphur dioxide. Natural colours such as anthocynins and betanin are used. Acid is an essential universal constituent of juice and the most commonly used acid is citric acid Oranusi et al., 2012). Fruit juices are easy to process and blended with other products (Bate et al., 2001).

Fruit juices contain sufficient nutrients that could encourage the growth of microorganisms. The most important factors which could encourage or limit the growth of microorganisms in juices includes pH, hygienic practice and storage temperature as well as the concentration of the preservative used (Okigbo and Obire, 2009; Tasnim et al., 2010; Odu and Adeniji, 2013). Several reports of illnesses due to the food borne diseases associated with the consumption of fruit juices at several places around the world have been reported (Chumber et al., 2007). Moreover, several factors can act as source of microbial contamination of the fruit juices such as use of unhygienic water for dilution, dressing with ice, prolonged preservation without refrigeration, unhygienic surroundings often with swarming houseflies and fruit flies and airborne dust (Tasnim et al., 2010; Odu and Adeniji, 2013). Various authors hve reported the presence of pathogens, namely, Escherichia coli, Salmonella spp., Shigella spp., and Staphylococcus aureus. Changes in pH may also promote the growth of pathogens (Food and Drug Administration. Tasnim et al., (2010) asserted that water used for juice preparation can be a major source of microbial contaminants including coliforms, faecal coliforms, faecal streptococci, etc.

The quality of fruit juices is strictly maintained in the developed countries and enforced under several laws and regulations. However, in many developing countries, although most of these are government agencies that regulate and control food and drug administration, small and medium entrepreneurs may not adhere strictly to microbiological safety and hygiene (Tasnim et al., 2010) and lack of strict adherence to standard operating procedures (SOP). Thus, the contamination of the juice becomes inevitable which could pose a public health risk.

1.1       AIM AND OBJECTIVES

The aim of this study is to evaluate the microbiological quality of Homemade drinks sold in Umuahia metropolis.

The objectives are;

1.     Determination of total aerobic bacteria plate count and total fungal count.

2.     Isolation and identification of bacteria and fungi contaminants from different vended Home made drinks sold within Umuahia.

3.     Determination of the percentage frequency distribution of the microbial isolates on the vended homemade drinks samples.

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