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KWAME NKRUMAH UNIVERSITY OF SCIENCE AND TECHNOLOGY, KUMASI COLLEGE OF SCIENCE DEPARTMENT OF FOOD SCIENCE AND TECHNOLOGY EVALUATION OF PROCESSING METHODS AND MICROBIAL ANALYSIS OF PITO (AN AFRICAN INDIGENOUS BEER), AT SELECTED PRODUCTION SITES IN GHANA BY JOHN-LEWIS ZINIA ZAUKUU (BSc Bio Analysis and Quality) A Thesis submitted to the Department of Food Science and Technology, College of Science in partial fulfilment of the requirements for the degree of Masters of Science in Food Science and Technology © 2015, Department of Food Science and Technology OCTOBER, 2015 DECLARATION I hereby declare that this submission is my own work towards the award of MSc. Food Science and Technology degree and that, to the best of my knowledge, it contains no material previously published by another person, nor material which has been accepted for the award of any other degree of the University, except where due acknowledgement has been made in the text. John-Lewis Zinia ZAUKUU ……………………… ………………… (Student) Signature Date Certified by: Prof. William O. ELLIS ……………………… ………………… (Supervisor) Signature Date Prof. Mrs Ibok ODURO ……………………… ………………… (Head of Department, Supervisor) Signature Date ii DEDICATION For giving me the best of all they had and withholding nothing good from me; I gladly dedicate this thesis report to my parents Mr. and Mrs. ZAUKUU and to my siblings. iii ACKNOWLEDGEMENT My utmost thanks go to the Lord of all wisdom, the source of all understanding in literature and science for good health and life To my supervisors Prof W.O Ellis and Prof. Ibok Oduro, I say thank you for believing in me to the end and for your strong motivation, attention and the encouragement i needed at various critical moments My sincere thanks also go to Ingrid Kuupole, William Appaw, Redeemer Agbolegbe, Damian Laryea and Christopher Asare for your support in making this work a success To my siblings, through you I understand better that blood is indeed thicker than water I say thank you to all my friends especially Benjamin Awaitey and Frances Wendy Grant for all the encouragement To all staff of the department of Food Science and Technology. I am indeed very grateful for your help iv TABLE OF CONTENT DECLARATION.......................................................................................................... ii DEDICATION.............................................................................................................iii ACKNOWLEDGEMENT .......................................................................................... iv TABLE OF CONTENT ............................................................................................... v LIST OF TABLES ....................................................................................................viii LIST OF FIGURES .................................................................................................... ix LIST OF PLATES ....................................................................................................... x LIST OF APPENDICES ............................................................................................ xi ABSTRACT ................................................................................................................ xii CHAPTER ONE .......................................................................................................... 1 INTRODUCTION........................................................................................................ 1 1.0 BACKGROUND OF STUDY ................................................................................. 1 1.1 PROBLEM STATEMENT ...................................................................................... 4 1.2 JUSTIFICATION .................................................................................................... 5 1.3 AIM AND OBJECTIVES........................................................................................ 5 1.3.1 AIM ....................................................................................................................... 5 1.3.2 OBJECTIVES ....................................................................................................... 5 CHAPTER TWO ......................................................................................................... 6 LITERATURE REVIEW ........................................................................................... 6 2.0 INTRODUCTION ................................................................................................... 6 2.1 Raw materials in indigenous Fermented foods ........................................................ 8 2.1.1 Millet ................................................................................................................... 10 2.1.2 Sorghum .............................................................................................................. 13 2.2 The Pito beverage .................................................................................................. 16 2.3 The Pito brewing process....................................................................................... 18 2.3.1 Malting ................................................................................................................ 18 2.3.2 Wort extraction ................................................................................................... 22 2.3.3 Fermentation, maturation and storage ................................................................. 24 2.4 Hazards associated with Pito ................................................................................. 26 2.4.1 Microbial hazards................................................................................................ 26 v 2.4.2 Heavy metals ....................................................................................................... 27 2.4.3 Mycotoxins ......................................................................................................... 28 2.4.3.1 Aflatoxins ......................................................................................................... 28 2.4.3.2 Health concerns associated with aflatoxin contamination ............................... 31 CHAPTER THREE ................................................................................................... 33 MATERIALS AND METHODS .............................................................................. 33 3.0 Research design ..................................................................................................... 33 3.1 Population of study ................................................................................................ 33 3.2 Data collection at the pito processing sites ............................................................ 34 3.3 Laboratory analysis and Handling procedures ....................................................... 35 3.4 Aflatoxin Determination ........................................................................................ 35 3.4.1 Extraction and clean-up protocol as described by AOAC (Official Method 999.07) and demonstrated in Plate 3.1 .............................................................. 35 3.4.2 AflaTest Affinity Chromatography (Plate 3.1) ................................................... 36 3.4.3 HPLC Quantification of Aflatoxins .................................................................... 37 3.5 Mould identification and mould count ................................................................... 38 3.5.1 Aseptic techniques in the laboratory ................................................................... 38 3.5.2 Microbial Sample Preparation ............................................................................ 39 3.5.3 Microbial sample preparation and enumeration ................................................. 39 3.5.4 Subculture of moulds .......................................................................................... 40 3.5.5 Identification of Molds ....................................................................................... 40 3.5.6 Coliform test ....................................................................................................... 40 CHAPTER FOUR ...................................................................................................... 42 RESULTS AND DISCUSSION ................................................................................ 42 4.0 Introduction ............................................................................................................ 42 4.1 Overview of interview and survey results ............................................................. 42 4.1.1 Processing method 1 ........................................................................................... 44 4.1.2 Processing method 2 ........................................................................................... 46 4.1.3 Processing method 3 ........................................................................................... 46 4.1.4 Processing method 4 ........................................................................................... 47 4.1.5 Processing method 5 ........................................................................................... 48 vi 4.3 Aflatoxin determination ......................................................................................... 56 4.4 Moulds Count........................................................................................................