ABSTRACT
This research investigated the effect of biochemical polymorphisms on performance traits in Nigerian indigenous chicken genotypes. The chickens were obtained from pedigree mating of Normal feathered, Frizzle feathered and Naked Neck cocks to Normal feathered, Frizzle feathered and Naked Neck hens respectively to produce F1 offspring. One hundred and fifty five chicks (37 Frizzle, 79 Normal and 39 Naked Neck) were measured for body weight (g), breast girth (cm) and tibia length (cm). At 20weeks, 5ml of blood was collected from wing vein of each chicken into heparinized tubes labeled according to its tag number for electrophoresis. Each bird was scored as either fast (AA), midway (AB) or slow (BB) according to the mobility on the cellulose-acetate paper for each of transferrin, haemoglobin and carbonic anhydrase. Data obtained were subjected to general linear model procedure of statistical analysis system (SAS, 2002)and significant means were separated using tukey honestly significant difference. Hardy Weinberg‟s equation was used to calculate genotypic and allelic frequencies and tested using chi-square (χ2). Each of the three biochemical markers distinguished into three polymorphic forms viz; AA, AB and BB. The Frizzle feathered had significant higher (P<0.05) weight (338.54g) than both the Normal feathered (319.59g) and Naked Neck (295.51g) from day old to 8weeks, with no significant differences (P>0.05) from 12 to 20 weeks with no consistent trend for tibia length and breast girth. The effect of sexual dimorphism was significant (P<0.05) with males having significantly higher body weight (1168.79g) than females (937.06g), breast girth, males (25.74cm) and females (24.24cm) and tibia length, males (9.59cm) and females (8.96cm). The genotypic frequencies of Transferrin (TfAA, TfAB and TfBB) had 8, 140 and 7 respectively, haemgolobin (HbAA, HbAB and HbBB), had 49, 56 and 50 respectively and carbonic anhydrase (CaAA, CaAB and CaBB) had 63, 79 and 13 respectively. The effect of the polymorphic forms on body weight (g), breast girth (cm), tibia length (cm) showed that the AA had significant higher (P<0.05) body weight (g) than AB and BB for; transferrin (1434.75g, 1047.11g, 1047.43g respectively) and haemoglobin (1296.43g, 1029.59g, and 884.46g respectively). The AA was also higher (P<0.05) than the AB and BB for breast girth (cm) and tibia length (cm) in the case of transferrin and haemoglobin. The carbonic anhydrase showed no consistent significant differences (P>0.05) between the different genotypes for body weight (g), breast girth (cm) and tibia length (cm). It could be concluded that body weight of the genotypes indicated Frizzle feather may be suitable for meat production. The effect of sexual dimorphism showed that the parameters were sex influenced. The high level of heterozygosity indicates that more of the heterozygotes adapted and survived better than the homozygotes. The effect of the polymorphic forms of the biochemical markers on body weight (g), breast girth (cm), and tibia length (cm) showed that transferrin and haemoglobin could be used for body weight selection, while carbonic anhydrase may not be used for selection of the measured parameters.
TABLE OF CONTENTS
TITLE PAGE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .i
DECLARATION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .ii
CERTIFICATION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .iii
DEDICATION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .iv
ACKNOWLEDGMENT. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .v
ABSTRACT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .vi
TABLE OF CONTENT. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .vii
LIST OF TABLES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . .viii
LIST OF PLATES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .xii
CHAPTER ONE
1.0 INTRODUCTION. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . .1
1.1 Justification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3
1.2 Hypotheses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5
1.3 Objectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5
CHAPTER TWO
2.0 LITERATURE REVIEW. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6
2.1 Origin and Domestication of chicken . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . .6
2.2 The use of Chicken in Research and Development . . . . . . . . . . . . . . . . . . . . . . . . . .6
2.3 Tropical Indigenous Animal Genetic Resources (TIAnGR) . . . . . . . . . .. . . . . . . . . .7
2.4 Management System and Distribution of Indigenous Chicken in Nigeria . . . . . . . . .8
2.5 Growth of Indigenous Chicken . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
2.6 Comparison between indigenous (Unimproved) and exotic (Improved) chickens. .10
2.7 Traits of Economic Importance in Chicken. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
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2.8 Linear Body Measurement in Chicken. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12
2.9 Biotechnology in Animal Production. . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . ..13
2.10 Electrophoresis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14
2.11 Polymorphism of Transferrin in Poultry. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16
2.12 Polymorphism of Haemoglobin in Poultry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .18
2.13 Polymorphism of Carbonic Anhydrase in Poultry . . . . . . . . . . . . . . . . . . . . . . . . . .20
2.14 Sexual Dimorphism in Chicken . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . .21
CHAPTER THREE
3.0 MATERIALS AND METHODS. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .23
3.1 Location of the Experiment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .23
3.2 Source of Experimental Animal, Breeding and Management . . . . . . . . . . . . . . . . .23
3.3 Feeding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .23
3.4 Mating Procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .26
3.5 Egg Collection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .26
3.6 Incubation of Eggs and Management of Chicks . . . . . . . . . . . . . . . . . . . . . . . . . . . .26
3.7 Data Collection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .28
3.7.1 Growth parameters or traits. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .28
3.7.2 Blood collection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .28
3.8 Preparation of Buffer and Other Solutions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .28
3.9 Electrophoresisof Blood Samples. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
3.10 Statistical Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .31
CHAPTER FOUR
4.0 RESULTS. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33
4.1 Body Weight and Body Linear Measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33
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4.2 Effect of Sex on the Body Weight(g), Breast Girth(cm) and Tibia Length(cm) of the whole Population of the Indigenous Chickens. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .35
4.3 Transferrin Genotype, Genotypic and Allelic Frequencies of the Frizzle, Normal and Naked Neck Indigenous Chicken of Nigeria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .37
4.4 Haemoglobin Genotype, Genotypic and Allelic Frequencies of the Frizzle, Normal and Naked Neck Indigenous Chickens of Nigeria. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .40
4.5 Carbonic Anhydrase Genotype, Genotypic and Allelic Frequencies of the Frizzle, Normal and Naked Neck Indigenous Chickens of Nigeria . . . . . . . . . . . . . . . . . . . . . . . . .43
4.6 Effect of Transferrin Genotype on Body Weight (g), Breast Girth (cm) and Tibia Length (cm) of the whole population of the Indigenous Chickens. . . . . . . . . . . . . . . . . . . .47
4.7 Effect of Transferrin Genotypes on Body Weight(g), Breast Girth(cm) and Tibia Length(cm) within the Frizzle Indigenous Chicken Population . . . . . . . . . . . . . . . . . . . . .49
4.8 Effect of Transferrin Genotypes on Body Weight (g), Breast Girth (cm) and Tibia Length (cm)within the Normal Indigenous Chicken Population . . . . . . . . . . . . . . . . . . . .51
4.9 Effect of Transferrin Genotypes on Body Weight (g), Breast Girth (cm) and Tibia Length (cm)within the Naked Neck Indigenous Chicken Population . .. . . . . . . . . . . . . . .52
4.10 Effect of Haemoglobin Genotype on the Body Weight (g), Breast Girth (cm)and Tibia Length (cm)of the wholePopulation of Indigenous Chicken. . .. . . . . . . . . . . . . . . .55
4.11 Effect of Haemoglobin Genotypes on Body Weight (g), Breast Girth (cm) and Tibia Length (cm)withinthe Frizzle Indigenous Chicken Population . . . . . . . . . . . . . . . . . . . . .57
4.12 Effect of Haemoglobin Genotypes on Body Weight (g), Breast Girth (cm) and Tibia Length (cm)withinthe Normal Indigenous Chicken Population . . . . . . . . . . . . . . . . . . . .59
4.13 Effect of Haemoglobin Genotypes on Body Weight(g), Breast Girth(cm) and Tibia Length withinthe Naked Neck Indigenous Chicken Population . . . . . . . . . . . . . . . . . . . . .61
4.14 Effect of Carbonic anhydrase Genotype on the Body Weight(g), Breast Girth(cm) and Tibia Length(cm) of the whole Population of the Indigenous Chickens. . . . . . . . . . .63
4.15 Effect of Carbonic Anhydrase Genotype on Body Weight (g), Breast Girth (cm) and Tibia Length (cm)withinthe Frizzle Indigenous Chicken Population . . . . . . . . . . . . . . . . 65
4.16 Effect of Carbonic Anhydrase Genotype on Body Weight(g), Breast Girth(cm) and Tibia Length(cm) withinthe Normal Indigenous Chicken Population . . . . . . . . . . . . . . . .67
4.17 Effect of Carbonic Anhydrase Genotype on Body Weight(g), Breast Girth(cm) and Tibia Length(cm) withinthe Naked Neck Indigenous Chicken Population . . . . . . . . . . ..69
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CHAPTER FIVE
5.0 DISCUSSION. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .71
5.1 Body Weight and Body LinearMeasurement . . . . . . . . . . . . . . . . . . . . . . . . . . . . .71
5.2 Effect of Sex on Body Weight (g), Breast Girth (cm) and Tibia Length (cm) on the whole Population of the Experimental Indigenous Chickens . . . . . . . . . . . . . . . . . . . . . . . .72
5.3 Transferrin Genotype, Genotypic and Allelic Frequencies of the Frizzle, Normal and Naked Neck Indigenous Chicken of Nigeria . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . .73
5.4 Haemoglobin Genotypic and Allelic Frequencies of the Frizzle, Normal and Naked Neck Indigenous Chicken of Nigeria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .75
5.5 Carbonic Anhydrase Genotype, Genotypic and Allelic Frequency of the Frizzle, Normal and Naked Neck Indigenous Chicken of Nigeria . . . . . . . . . . . . . . . . . . . . . . . . . .76
5.6 Effect of Transferrin Genotype on the Body Weight (g), Breast Girth (cm) and Tibia Length (cm) of the whole population of indigenous chicken and within the individual populations of Frizzle, Normal and Naked Neck. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .76
5.7 Effect of Haemoglobin Genotype on the Body Weight (g), Breast Girth (cm) and Tibia Length (cm) of the whole population of indigenous chicken and within the individual populations of Frizzle, Normal and Naked Neck . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .78
5.8 Effect of Carbonic Anhydrase Genotype on the Body Weight, Breast Girth and Tibia Length of the whole population of indigenous chicken and within the individual populations of Frizzle, Normal and Naked Neck . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..79
CHAPTER SIX
6.0 SUMMARY, CONCLUSION AND RECOMMENDATIONS. . . . . . . . . . . . . . . .80
6.1 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .80
6.2 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . .80
6.3 Recommendations . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . .82
REFERENCES. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
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CHAPTER ONE
1.0 INTRODUCTION
The local breeds of chicken also referred to as indigenous chickens are genetically adapted to harsh environment facing limited resources and severe challenges from climatic conditions, pathogens and predators (Mwacharo et al., 2005; Sorenson, 2009). They are often utilized for several purposes simultaneously (Sorenson, 2009; Adeleke et al., 2011a), and possess both superior levels of genetic diversity/variation relative to commercial breeds (which have been selected for a particular performance traits) and they have unique traits of valuable local adaptations (Sorenson, 2009). Some local chickens have special characteristics of potential interest to commercial breeders. Therefore, indigenous chickens could be genetic source for future breeding strategies (Horst, 1999).
Genetic or protein polymorphism results from variations in the proteins or specifically in the amino acids for which the same genes code in different individuals, strains, breeds (Rege and Okeyo, 2006; Kwaga, 2006). This is the occurrence of two or more discontinuous forms of protein in a species/population in such a proportion that the rarest phenotype which has a frequency of more than 0.1 percent cannot be maintained merely by recurrent mutation (Das and Deb, 2008). Protein polymorphism can be used to map (locate) genes such as those causing a disease, for economic traits, selection of superior animals for breeding purposes (Akpa et al., 2011) and they can help match two samples of deoxyribonucleic acid (DNA) to determine if they come from the same source. In general, this variation in proteins can be used for the study of genetic diversity within a population‟s gene pool by applying two approaches viz; protein electrophoresis and protein
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immunology, bearing in mind that the basic principle behind electrophoretic mobility of enzymes as well as other proteins is mobility across gels, which denotes differences in allelic groups responsible for amino acid variations in the protein (Rege and Okeyo, 2006).
Transferrin is a polymorphic blood protein found in the serum and in milk of animals (Steppa et al., 2009), it is synthesized at high levels in an egg laying bird and is secreted as a major component of the egg white gene (Lee et al., 1980). Transferrin is characterized by the highest heterogeneity among all polymorphic blood proteins (Steppa et al., 2009). Das and Deb (2008) reported that three types of alleles viz TfA, TfB and TfC were observed in chicken on the gel electrophoresis separation. The chickens with type „TfB‟ have an advantage in egg production over the chickens with TfA. The effect of heterozygous transferrin (TfBC) appears to have significant variability in the fertility, hatchability and egg production (at least 90 days‟ production) and chicken with TfA have delayed sexual maturity while the chicken with the TfB has earlier age at sexual maturity.
Haemoglobin is an erythrocyte pigment, conjugated globins-prosthetic group, and also a polymorphic protein (Das and Deb, 2008). It carries oxygen and carbon dioxide (Das and Deb, 2008; Steppa et al., 2009) and participates in maintenance of proper blood reaction (Steppa et al., 2009). Das and Deb (2008), reported three haemoglobin polymorphic forms, controlled by two autosomal alleles A1 and A2 with genotypic frequencies as 0.96 : 0.04 for White Leghorn, 1.00 : 0.00 for local fowl, 1.00 : 0.00 for Guinea fowl and 0.85 : 0.15 for Japanese quail. Haemoglobin polymorphism affects growth rate, hatchability and susceptibility of chicken to Marek‟s disease (Das and Deb, 2008). Steppa et al. (2009), however, reported that there were two codominant autosomal alleles, found in the beta-haemoglobin chain in sheep.
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Carbonic anhydrase was not directly reported to affect weight of chicken, but, it was recognized to affect the transportation and utilization of haemoglobin in the body of chickens (Das and Deb, 2008), while haemoglobin is known to affect growth rate in chickens. The effect of the carbonic anhydrase on heamoglobin could therefore indirectly translate into effect on the growth rate of chickens.
1.1 Justification
In order to measure genetic diversity within and between breeds/strains, the field of molecular biology, particularly the application of molecular markers to study genetic diversity, has evolved very rapidly since the mid-1960s. The dominance of protein electrophoretic approaches to population genetics and evolutionary biology was, in the late 1970s, replaced by DNA analysis, primarily through the use of restriction enzymes, and in the 1980s by mitochondrial DNA analyses and DNA fingerprinting approaches (Rege and Okeyo, 2006). More recently, the introduction of PCR-mediated (polymerase chain reaction-mediated) DNA genotyping/sequencing has provided the first rapid and easy access to the ultimate genetic data (Miao et al., 2013). Although DNA based technologies are now the methods of choice, it would be a mistake to conclude that DNA markers provide the ultimate solution. Several alternative assays, such as protein/allozyme polymorphisms (biochemical), remain tremendously useful, especially in developing countries, because of their utility, ease, cost and amount of genetic information accessed or simplicity of data interpretation (Rege and Okeyo, 2006).
To date, characterization of quantitative traits in livestock, especially in Nigeria has been based majorly on phenotype or on estimated breeding values (EBV) derived from
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phenotype without proper knowledge on how many genes are actually playing effective role on the trait or knowing the specific effect of the individual genes, and thus the genetic foundation of traits is usually treated as “hidden actor” (Naqvi, 2007).
Transferrin could be obtained in blood serum, plasma and eggs of poultry (Frelinger, 1972; Jaayid et al., 2011) and the polymorphic forms of it affects sexual maturity, while sexual maturity of an animal is a function of its growth, and it therefore has implication to detect performance in terms of weight gain. Haemoglobin polymorphism on the other hand has been directly linked to affect the growth rate and hatchability of chickens. The polymorphic nature of both transferrin and haemoglobin is said to be responsible for the differences observed in performances of the birds and could serve for evaluation and selection for improvement in genetic performance (Das and Deb, 2008). Carbonic anhydrase participate in the regulation of ion, water and acid-base balance, while some members of carbonic anhydrase gene family have been suggested to promote cell proliferation and act as trophic/growth factor (Karhuma, 2002). Therefore, by applying different biochemical – genetic markers like transferrin, haemoglobin, carbonic anhydrase; the individual animal and the population as a whole could be genetically well characterized in relation to their performance (Jaayid et al., 2011).
This study therefore, was conducted to investigate the effect of biochemical polymorphisms on performance traits in indigenous chicken genotypes in order to know the basic genetic activities which contribute to the differences observed in terms of weight in the performance of Nigerian local chicken genotypes especially as research of this kind is limited in the country.
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1.2 Hypotheses: This study was carried out to investigate the following null and alternative hypotheses:
H0: Biochemical Polymorphic form of transferrin, haemoglobin and carbonic anhydrase in Nigerian local chickens of Normal feathered, Frizzle feathered and Naked Neck have no effect on performance traits.
HA: Biochemical Polymorphic form of transferrin, haemoglobin and carbonic anhydrase in Nigerian local chickens of Normal feathered, Frizzle feathered and Naked Neck have effect on performance traits.
1.3 Objectives
This study was designed to address the following:
i. To examine the differences in weight between and within the Nigerian local chicken Normal feathered, Frizzle feathered and Naked Neck.
ii. To calculate the genotypic and allelic frequencies of transferrin, haemoglobin and/or carbonic anhydrase of the different genetic groups and of the whole population combined.
iii. To determine the effect of the polymorphic forms of transferrin, haemoglobin and carbonic anhydrase on growth performance of Normal feathered, Frizzle feathered and Naked neck Nigerian indigenous chickens.
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