MICROORGANISMS ASSOCIATED WITH KUNU ZAKI AND THEIR RELEVANCE IN PUBLIC HEALTH

ABSTRACT

This study evaluated kunu for microorganisms of public health important in Umuahia metropolis. A total of 19 bacterial and fungal species were isolated using the pour plate technique. The bacterial isolates identified using morphological and biochemical characterization included; Escherichia coli, Staphylococcus aureus, Bacillus spp, Lab plantarum, Klebsiella species, and Streptococcus spp. while the fungal species included; Aspergillus niger, Aspergillus flavus and Saccharomyces cerevisiae. The presence of these pathogens even in small numbers could render a beverage unsuitable for human consumption. The viable microbial counts on the kunu zaki drink revealed that the count ranges from 1.1x105 to 7.5x105, while the fungi count ranges from 1.0x105 to 5.5x105­­. In the evaluation of kunu zaki drink for microorganisms of public health importance, among the bacteria isolates, E.coli 4(21.0 ) had the highest level of occurrence followed by Klebsiella spp and Bacillus spp respectively at 3(15.7 ) each, while the least occurring isolates was Streptococcus spp with percentage at 1(5.3%). The high microbial load in this study may be attributed to poor sanitary conditions during production of the kunun drink.  Therefore, unclean water used in the preparation of the drink could lead to contamination, also the use of improperly washed hands after going to the toilet in the preparation of the drink could also be a source of contamination. Ingestion of some of these organisms is capable of causing human illnesses such as pneumonia, food-borne intoxification, diarrhea, gastroenteritis, among others. Therefore, there is need to maintain adequate sanitary conditions during the preparation, and handling of kunun drink to prevent contamination.

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                                                                                                       5

2.1       Kunun-Zaki as a Beverage                                                                                         5

2.2       Microbial Evaluation of Kunun-Zaki Prepared and Sold in Bauchi Metropolis      6

2.3       Microbial Quality of Kunun-Zaki Sold in Eiyenkorin, Kwara State, Nigeria            7

2.4       Microbiological Analyses of Kunu Drinks Locally Produced and Sold in Calabar           9

2.4.1    Determination of Total Bacterial and Fungal Counts                                                9

2.4.2    Purification of Microbial Isolates                                                                               10

2.4.3    Identification of Bacterial and Fungal Isolates                                                          10

2.5       Microbiological Analyses of Hawked Kunun and Zobo

            Drinks within Lautech Campus, Ogbomoso, Oyo State, Nigeria                                    11

2.6       Microbiological Quality of Kunu Drink Sold in Some Major

            Markets of Yenagoa   Metropolis, Nigeria                                                                12

CHAPTER THREE

3.0       Materials and Methods                                                                                               15

3.1       Collection of Samples                                                                                                15

3.2       Sterilization of Materials                                                                                            15

3.3       Sample Preparation and Isolation of Micro Organisms                                             15

3.3.1    Determination of Total Viable Plate Counts                                                              15

3.3.2    Determination of Total Fungal Plate Count                                                               15

3.4       Purification of Isolates                                                                                               16

3.5       Identification of Bacteria Isolates                                                                               16

3.6       Gram Staining                                                                                                            16

3.7       Biochemical Test                                                                                                        17

3.7.1    Catalase Test                                                                                                               17

3.7.2    Indole Test                                                                                                                  17

3.7.3    Citrate Utilization Test                                                                                               17

3.7.4    Hydrogen Sulphide (H₂S) Production Test                                                                17

3.7.5    Starch Hydrolysis                                                                                                       18

3.7.6    Motility, Indole, Urease (MIU)                                                                                  18

3.7.7    Coagulase Test                                                                                                           19

3.7.8    Oxidase Test                                                                                                               19

3.8       Identification of Fungal Isolates                                                                                 19

3.8.1    Wet Preparation                                                                                                          19

3.8.2    Colonial Morphology                                                                                                 19

CHAPTER FOUR

4.0       Results                                                                                                                        20

CHAPTER FIVE

5.0       Discussion, Conclusion and Recommendation                                                          26

5.1       Discussion                                                                                                                   26

5.2       Conclusion                                                                                                                  29

5.3       Recommendation                                                                                                       30

            References

LIST OF TABLES

CHAPTER ONE

1.0       INTRODUCTION

Kunun-zaki drink is a locally prepared indigenous non-alcoholic beverage which is widely produced and consumed in large quantity in Nigeria, especially the northern part of the country (Akoma et al., 2006). This local beverage is consumed in both wet and dry season but more in the dry season because of its optimal thirst quenching abilities. Kunu is consumed any time and by both adults and children as breakfast drink, food complement, as a refreshing drink usually used to entertain visitors.

It is used to stimulate appetite and is commonly served at social gatherings and according to Onuorah et al., (2007), the non-alcoholic nature of the drink makes it to be readily consumed by Christians and Muslims alike as a substitute for alcoholic ones. The drink is mostly consumed within 20-35 h of its production due to its poor keeping quality (Akoma et al., 2006). This drink is not expensive because the grains and other ingredients used for its production are locally sourced and are mostly grown within the savannah region and almost throughout the year. The packaging materials are also available, cheap and easily affordable. No elaborate equipment and expertise are required for its production (Oluwalana and Adedeji, 2013). It is also very nutritious and of medicinal value. Kunun-zaki has been reported to be rich in vitamins, minerals, carbohydrates and proteins (Efiucwere et al., 2005). Kunu zaki processed from Sorghum grain has been reported to contain 11.6% protein, 3.3% fat, 1.9% ash and 76.8% carbohydrate and arrays of amino acid. The different types of kunu in Bauchi are: kunu zaki, kunu gyade, kunun tsamiya, kunun baule, kunun jiko amshar and kunu gayamba as they are called in different tribes of Bauchi state.

The drink is produced from fermented millet, sorghum (guinea corn) and maize in decreasing order of preference. In some culture, the grains are used in composite form especially millet and sorghum (guinea-corn) in a ratio of 1:2 w/w (Abega, 2007). It is sweetened with honey and sugar together with small quantities of sweet potatoes and spices (Ginger or black pepper). Its processing is mostly done by women using simple household equipment and utensils. The processed kunun zaki is usually packaged and sold in 50ml to 1 litre plastic bottles and at times tied in some disposable polythene bags or in bulk inside large containers and distributed under ambient temperature or cooled in a refrigerator where available.

In developing nations like Nigeria, it has not been possible to have control over the processing and sales of this beverage. Kunu zaki has high moisture content; its method of production is crude and is packaged under unhygienic condition. It is also highly consumed due to the cost of other non-alcoholic drinks (Abega, 2007). The drink is hawked in the motor parks, school premises and in market places where it is exposed to contamination. These and other factors predispose kunu zaki to microbial contamination.

The production of kunu drink is still at village technology level where procedures and materials or ingredients used are always not standardized. In general, the process of production involves steeping the grains for 6-24 hours, wet milling with spices, wet sieving and partial gelatinization of the slurry (Oranusi et al., 2003). This is followed by addition of sugar and spices depending on the producer, such as ginger (Zingiber officimals) and alligator pepper (Afromonium melegueta) or red pepper (Capsicum species), or black pepper (Piper guineense). These serve as flavor and taste improver which is not quantified (Gaffa et al., 2002). The consumption of locally made kunu drinks by both old and young people including market women and children at home pose serious health threats because these products are subject to microbial contamination as producers do not adhere to standard methods of preparation. They also lack information on the microbiological safety of ready-to-eat beverages and their health implications (Oranusi et al., 2003).

Studies have shown that kunun plays a vital role in the dietary pattern of the people in developing countries like Nigeria (Ogbonna et al., 2011). Mbachu et al., (2014) reported that kunun zaki contains 0.3% protein, 1.0% fat, 1.52% ash, 12.2% carbohydrate and 8.9 mg of vitamin C per 100 ml. More so, some studies have demonstrated the health benefits of kunun to include but not limited to reduction of risk associated with diabetes, lowering of blood cholesterol, prevention of formation of blood clot and helps to fight against cancer (Ofudje et al., 2016). Traditionally, the production procedure of kunun varies depending on the taste and cultural habits of the consumers, thus leading to variation in quality and stability. Aboh and Oladosu (2014) reported that production methods of kunun-zaki are crude, ingredient concentrations are neither quantified nor standardized, but instead preparation is largely a matter of family tradition. There is possibility of contamination at various stages of unit operations. The production is usually carried out under unhygienic conditions which may predisposed it to many pathogens of public health importance. Ayandele, (2015) noted that the spices usually added being agricultural commodities may contain high level of microbial impurities, which can serve as a source of contamination. Umaru et al., (2014) reported that locally prepared beverages may serve as vehicles for zoonotic and food-borne diseases or pathogens; such as Staphylococcosis, Salmonellosis, Brucellosis, Tuberculosis, Listeriosis, Shigellosis, E. coli, etc. Thus, this study was undertaken to assess the microbial quality of kunun drink prepared and sold in Eiyenkorin, Kwara State, Nigeria. Findings from this study can assist government regulatory agencies in monitoring the quality of kunun and other local beverages.

1.1       AIM AND OBJECTIVES

To evaluate Kunun-zaki drink (a locally prepared indigenous non-alcoholic beverage drink) for microorganisms of public health importance, while the specific objectives are;

Ø  To isolate and identify the microorganisms associated with Kunun-zaki drink sold within Umuahia

Ø  To determine the percentage occurrence of various isolates from Kunun-zaki drink sold within Umuahia

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