SEX IDENTIFICATION OF NIGERIAN INDIGENOUS FRIZZLE FEATHERED AND NAKED NECK CHICKS USING VENT SEXING AND MOLECULAR TECHNIQUE

ABSTRACT

Sex  determination of  Nigerian indigenous  frizzle  feathered and naked neck chicks  has  been a difficult  task especially  due  to the  monomorphism  and high variation in  the  feather  colour  of  this indigenous  chickens  using  the  feather  sexing method. Thus, the  vent  sexing method is  cheap  and easy  to  carry  out  depending  on  the  level  of  expertise  and  the  molecular  sexing  technique  was developed from  recent  studies  to accurately  sex  birds  using  a  PCR  based sex  determination protocol  targeting  the  CHD-W  and  CHD-Z  genes  in the  chicken’s  chromosome  Z  and chromosome W respectively.  The  percentage  accuracy  of  vent  sexing  method obtained in  this  study  were  66.7% and 77.8%  for  the  frizzle feathered  and  naked  neck  chicks  respectively.  The  result  observed  from this  study  show  that  the  vent  sexing  method is  easy  and  cheap to carry  out  but  relatively  low  in accuracy  and carrying  out  the  PCR  method  was  expensive  and inefficient  due  to the ineffectiveness of  the  DNA  premix  used in this  study. 

TABLE OF CONTENTS

CONTENT PAGE

Title Page …………………………………………………………………...…………………i

Certification ………………………………………………………...…………….…..…………. ii

Dedication……………………………………………………………………...………..………..iii

Acknowledgement…………………………………………………...………………………….. iv

Abstract……………………………………………………………………………………………v

Table of contents………………………………………………………….………………………vi

List of Tables……………………………………………………………………...…………...….ix

List of Figures…………………………………………………………………………………….ix

List of Plates……………………………………………………………………….……………..ix

CHAPTER ONE

1.0 INTRODUCTION……………………………………………………………...…………1

1.1 Justification…………………………………………………………..……………4

1.2 Objectives………………………………………………………..………………..4

1.2.1 Broad Objective………………………………………………..………….4

1.2.2 Specific Objectives……………………………………………….....…….4

CHAPTER TWO

2.0 LITERATURE RIVIEW

2.1 The Nigerian Indigenous chicken……………………………………..…………..5

2.2 The Frizzle feathered chicken……………………………………….…………….6

2.3 The Naked neck chicken……………………………………………….………….7

2.4 Sex Identification in chicken………………………………………...……………8

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2.5 Vent sexing of chicken……………………………………...……………………..9

2.6 Molecular techniques for sexing of chicken…………………………..…………10

2.6.1 DNA sampling………………………………………………………..….10

2.6.2 DNA storage……………………………………………..………………11

2.6.3 DNA extraction methods……………………………...…………………11

2.7 Sexing techniques based on Polymerase chain reaction (PCR)……………….…11

2.7.1 Reagents in regular PCR…………………………………………………12

2.7.2 PCR process and regular temperature profiles……………………...……13

2.8 Electrophoresis………………………………………………………...…………15

2.8.1 Gel electrophoresis………………………………………….……………15

2.9 CHD gene - based sex identification……………………….……………………16

CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Experimental site……………………………………………………..………….18

3.2 Experimental birds……………………………………………...………………..18

3.3 Management of birds…………………………………………………………….18

3.4 Semen collection and Artificial insemination……………………………………18

3.5 Egg collection and Incubation……………………………………...……………19

3.6 Collection of chicks and Tagging…………………………….………………….19

3.7 Vent sexing……………………………………………………...……………….19

3.8 Collection of Genomic DNA Samples……………………………………..…….20

3.9 Genomic DNA Extraction…………………………………………..……………20

3.10 Genomic DNA quantity and quality check…………………..…………………..21

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3.11 Polymerase Chain Reaction (PCR) amplification……………….……………….21

3.12 Electrophoresis……………………………………..…………………………….22

3.13 Data Collection………………………………………..…………………………22

3.14 Verification exercise……………………………………………………………..23

3.15 Statistical Analysis……………………………………………………………….23

CHAPTER FOUR

4.0 RESULTS AND DISCUSSION

4.1 Results…………..………………………………………………………………..25

4.2 Discussion…………………….………………………………………………….34

CHAPTER FIVE

5.0 CONCLUSION AND RECOMMENDATION

5.1 Conclusion……………………………………………………...………………..35

5.2 Recommendation………………………………………….……………………..35

REFERENCES……………………………….………………………………………………….37