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ABSTRACT

This study was undertaken to evaluate some selected productive traits and relationship between three genetic groups of cattle (Bunaji, Friesian X Bunaji cross and Sokoto Gudali) using blood biochemical polymorphism. A total of 150 cows consisting of 50 per genetic group was randomly sampled from the (NAPRI) cattle herd and used for the study. The selected productive traits evaluated include body height: (BW) in Kg, body length in cm (BL) height at withers (HW) in cm, chest width (cw), heart girth (HG), rump width ((Rumw) teat length in cm (TL), rear udder height (RUH) udder circumference (UC), total milk yield (TY), average daily milk yield (ADY) and lactation length in days (LL). Blood biochmeical parameters, namely Haemoglobin, Transferrin and carbonic anhydrase were evaluated for genotypic and alletic frequencies of these blood protein polymorphism and their impacts on productive traits, correlated analysis of the measured trait, principal component analysis of the variables, stepwise linear regression and multivariate analysis involving discriminant genetic distance and classification components between breeds were all computed. Results showed that BW and other measured traits differed significantly (P<0005) among the genetic groups BW and BL were higher in the Friesian X Bunaji than the Bunaji and Sokoto Gudali which showed no significant (P>005) difference for these traits. The haemorglobin1ocus revealed overall allelic frequencies of 0.44 and 0.13 for HbAA and HbBB respectively. Low frequencies of alleles (0.20, 0.20 and 0.40) were observed in the Bunaji, Friesian X Bunaji and the Sokoto Gudali respectively. In the transferrin locus, only the A and B alleles were observed with overall frequencies of 0.16 and 0.09 respectively. Low genotypic frequency of 0.04 for AA was obtained in the Sokoto Gudali while the Bunaji and the Friesian X Bunaji recorded frequencies of 0.30 and 0.12 respectively. In the carbonic anhydrase locus, only the F and S alleles were observed with the overall frequencies of 0.17 and 0.23 respectively. Both the Tranferrin and Carbonic Anhydrase locus were not in Hardy-Weinberg equilibrium for the studied population. Significant (p<0.05 differences were obtained in all body and milk production traits measurements of the three studied population indicating clear genetic group distinction. It was noted that the Sokoto Gudali was superio to the Bunaji in most of these traits though the Sokoto Gudali is a relatively poor milker than the Bunaji. Study of blood protein polymorphism and productivity indicated significant (p<0.05) influence of the haemoglobin, transferrin and carbonic anhydrase on both body and milk
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production traits. Correlated studies were observed to be very significant between variables in the pooled analysis by majorly insignificant for individual genetic groups, also estimates were generally low though ranging from 0.87 – 0.84. Principal component analysis observed to show factors ranging from 3 in the pooled data to 5 in the Bunaji and 6 in the Friesian X Bunaji and Sokoto Gudali genetic groups. Generally communalities ranged from 0.31 – 099 whils proportion of variance accounted for by factors were 47% in the pooled and Bunaji and 58% in the cross breed and Sokoto Gudali. Multivariate analysis indicated TY, CW, ADY, RUH, LL, Rumwi, UC, TL and HG as the most discriminating variable among the genetic groups. Their respective partial R2 and F values were (0.84, 0.58, 0.22, 0.150.13, 0.09, 0.08, 0.15 and 0.04) and (2278.64, 613.22, 123.85, 81.12, 65.12, 43.02, 37.10, 79.68 and 16.52) with high significant value of (p<0.0001). Genetic distance among the genetic groups revealed high Mahalanobis value (3.94 – 4.95) between the Bunaji and Sokoto Gudali and Friesian X Bunaji and Sokoto Gudali. The Highest proper classification (84%) was in the Sokoto Gudali and it indicated greater genetic group homogeneity. However, multivariate studies clearly indicated manifestation of gene introgression across genetic groups through indiscriminate cross breeding. Perhaps as evidenced by the high levels of misclassification in the Bunaji, Friesian X Bunaji and Sokoto Gudali. There is a need for generic study using protein and DNA microsatellite makers to compliment the results arisen from morphometric differentiation of the two most popuous Nigerian breeds of cattle in the NAPRI herd.

 

 

TABLE OF CONTENTS

Contents page
Title Page ……………………………………………………………………………………………………………………………… i
Cover Page………………………………………………………………………………………………………………………………………….ii
DECLARATION …………………………………………………………………………………………………………………. iii
CERTIFICATION ………………………………………………………………………………………………………………. iv
DEDICATION ……………………………………………………………………………………………………………………… v
ACKNOWLEDGEMENT …………………………………………………………………………………………………….. vi
ABSTRACT ……………………………………………………………………………………………………………………….. vii
TABLE OF CONTENTS ……………………………………………………………………………………………………… ix
CHAPTER ONE …………………………………………………………………………………………………………………… 1
1.0 INTRODUCTION ………………………………………………………………………………………………………. 1
1.1 Justification of the Study ………………………………………………………………………………………………. 3
1.2 Objectives of the Study …………………………………………………………………………………………………. 4
1.3 Hypotheses of the Study ………………………………………………………………………………………………… 4
CHAPTER TWO………………………………………………………………………………………………………………….. 5
2.0 LITERATURE REVIEW ……………………………………………………………………………………………… 5
2.1 Biotechnology Options for Improving Livestock Production …………………………………………….. 5
2.1.1 Biochemical polymorphism …………………………………………………………………………………………… 6
2.1.2 Genetic control on biochemical polymorphism …………………………………………………………………. 6
2.2 Electrophoresis…………………………………………………………………………………………………………….. 7
2.2.1 Protein electrophoresis …………………………………………………………………………………………………. 7
2.2.2 Cellulose acetate electrophoresis ……………………………………………………………………………………. 8
2.2.3 Sodium dodecyl sulphate poly acrylamide gel electrophoresis …………………………………………… 10
2.3 Blood Protein and Enzyme Types ………………………………………………………………………………… 14
2.3.1 The haemoglobins ……………………………………………………………………………………………………… 14
2.4 Haemoglobin Types and Economic Traits: ………………………………………………………………….. 20
2.4.1 Haemoglobin types and production traits. ………………………………………………………………………. 20
2.4.2 Haemoglobin types and adaptation traits: ………………………………………………………………………. 22
2.4.3 Haemoglobin physiological function …………………………………………………………………………….. 22
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2.5 The Transferrins ………………………………………………………………………………………………………… 23
2.5.1 Transferrin transport mechanism ………………………………………………………………………………….. 23
2.5.2 Structure of TRANSFERRIN …………………………………………………………………………………….. 24
2.5.4 Role of transferrin in disease control……………………………………………………………..25
2.5.5Transferrin types and economic traits: ……………………………………………………………………………. 25
2.5.6 Transferrin types with age at first lambing. …………………………………………………………………….. 26
2.5.7 Transferrin types with lambing interval …………………………………………………………………………. 26
2.5.8 Physiological function of transferrin ……………………………………………………………………………… 27
2.6 Carbonic Anhydrase. ………………………………………………………………………………………………….. 27
2.6.1 Carbonic anhydrase in health and disease: ……………………………………………………………………… 28
2.6.2 Physiological function of carbonic anhydrase: ………………………………………………………………… 28
2.7 Genetic Types of Carbonic Anhydrase. …………………………………………………………………………. 29
2.8 PERFORMANCE TRAITS …………………………………………………………………………………………. 29
2.8.1 Morphometric measures in cattle ………………………………………………………………………………….. 29
2.8.2 Udder characteristic and milk production ………………………………………………………………………. 30
2.10 Genetic Introgression and Breeding Herds Morphometric Attributes …………………………………… 32
CHAPTER THREE…………………………………………………………………………………………………………….. 35
3.0 MATERIALS AND METHODS ………………………………………………………………………………….. 35
3.1 Location of the experiment ………………………………………………………………………………………….. 35
3.2 Animals and Management …………………………………………………………………………………………… 35
3.3 Bodyweights ………………………………………………………………………………………………………………. 36
3.4 Body Linear Measurement ………………………………………………………………………………………….. 36
3.5 Udder Measurements ………………………………………………………………………………………………….. 36
3.6Milk Yield Characteristics ……………………………………………………………………………………………. 37
3.7 Cellulose Acetate Electrophoresis ………………………………………………………………………………… 37
3.7.1 Blood collection and sample preparation ……………………………………………………………………….. 37
3.8Sample Preparation …………………………………………………………………………………………………….. 38
3.8.1 Blood haemolysates and plasma collection …………………………………………………………………….. 38
3.9Experimental Procedure: …………………………………………………………………………………………….. 38
3.9.1 Gel soaking ………………………………………………………………………………………………………………. 38
3.9.2Sample loading ………………………………………………………………………………………………………….. 39
3.9.3 Gel running ………………………………………………………………………………………………………………. 39
3.9.4 Gel staining………………………………………………………………………………………………………………. 39
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3.9.5 Gel and bands scoring ………………………………………………………………………………………………. 40
3.9.6 Estimation of the gene and genotypic frequency ……………………………………………………………… 40
3.10 Electrophoretic Conditions ………………………………………………………………………………………… 42
3.10.1 Measurement of genetic distance. ……………………………………………………………………………….. 43
3.11 Statistical Analysis ……………………………………………………………………………………………………. 43
3.11.1 Model for the analysis was a factorial anova design as illustrated below: …………………………… 43
3.11.2 Correlation analysis………………………………………………………………………………………………….. 43
3.11.3 Principal component analysis procedures……………………………………………………………………… 44
CHAPTER FOUR ………………………………………………………………………………………………………………. 47
4.0. RESULTS …………………………………………………………………………………………………………………. 47
4.1 Genotype and Gene Frequencies of Blood Proteins among Three Genetic Groups ……………. 47
4.1.1 Genotype and gene frequencies of haemoglobin among three genetic groups ……………………….. 47
4.1.2 Genotype and gene frequencies of transferrin among three genetic groups …………………………… 47
4.2 Performance of three Genetic groups in Morphometric and selected Milk Productio ……….52
traits ………………………………………………………………………………………………………………………………. 52
4.3 Effect of Haemoglobin Types on Morphology and Milk Production ………………………………… 52
4.4 Influence of Transferrin types on Morphology and Milk Production ……………………………….. 53
4.5 Influence of Carbonic Anhydrase on Morphology and Milk Production ………………………….. 58
4.6 Effect of Genetic Groups and Haemoglobin forms on Morphology and Milk Traits ………….. 60
4.7 Effect of three Genetic groups and Transferrin forms on Morphology and Milk Traits …….. 62
4.8 Effect of three Genetic Groups and Carbonic Anhydrase forms on Morphology and Milk Traits ………………………………………………………………………………………………………………………… 64
4.9 Correlated Studies ……………………………………………………………………………………………………… 64
4.9.1 Correlation of growth and milk traits for all genetic groups ………………………………………………. 64
4.9.2Correlation of growth and milk production traits in the Bunaji……………………………………………. 65
4.9.3Correlation of growth and milk production traits in the Friesian X Bunaji…………………………….. 65
4.9.4Correlation of growth and milk production traits in the Sokoto Gudali…………………………………. 65
4.10 Multivariate Principal Component Analysis ………………………………………………………………… 66
4.10.1Principal component analysis of morphometric traits for all genetic groups ………………………… 66
4.10.2Principal component analysis of morphometric traits in the Bunaji ……………………………………. 66
4.10.3Principal component analysis of morphometric traits in the Friesian X Bunaji …………………….. 71
4.10.4Principal component cnalysis of torphometric traits in the Sokoto Gudali …………………………… 71
4.11 Stepwise Linear Regression Predictor for Total Milk Yield …………………………………………… 71
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4.12 Discriminant Analysis of Selection Traits ……………………………………………………………………. 72
4.13 Genetic Distance and Classification into Genetic Groups ……………………………………………… 72
CHAPTER FIVE …………………………………………………………………………………………………………….. 82
5.0 DISCUSSION …………………………………………………………………………………………………………….. 82
5.1 Genotype and Gene Frequencies of Blood Proteins among three Genetic Groups ……………… 82
5.1.1 Genotype and gene frequencies of haemoglobin among three genetic groups ……………………….. 82
5.1.2Genotype and gene Frequencies of Transferrin among genetic groups …………………………………. 83
5.1.3Genotype and gene Frequencies of Carbonic Anhydrase among genetic groups …………………….. 84
5.2 Performance of Genetic groups in Morphometric and Selected Milk Production Traits…….. 84
5.3 Blood Polymorphism and Productivity …………………………………………………………………………. 86
5.3.1 Effect of haemoglobin types on morphology and milk production ……………………………………… 86
5.3.2 Effect of transferrin types on morphology and milk production………………………………………….. 87
5.3.3Effect of carbonic anhydrase types on morphology and milk production. ……………………………… 88
5.4 Correlated Relationship Studies …………………………………………………………………………………… 88
5.5 Principal Component Analysis of Morphometric Traits …………………………………………………. 90
5.6 Stepwise Linear Regression Predictor for Total Milk Yield …………………………………………….. 91
5.7 Multivariate Analysis of Selection Traits ………………………………………………………………………. 91
5.8 Genetic Distance Study ……………………………………………………………………………………………….. 92
CHAPTER SIX…………………………………………………………………………………………………………………… 94
6.0 SUMMARY, CONCLUSION AND RECOMMENDATIONS. ………………………………………… 94
6.1 Summary …………………………………………………………………………………………………………………… 94
6.2 Conclusion …………………………………………………………………………………………………………………. 96
6.3 Recommendations ………………………………………………………………………………………………………. 97
REFERENCES…………………………………………………………………………………………………………………….. 98

 

 

CHAPTER ONE

 

1.0 INTRODUCTION
Genetic characterization of indigenous breeds and their biochemical traits form one important component in a strategy to expand food production. Several studies have been published on various livestock species in Nigeria (Das and Deb, 2008). The lindigenous breeds have received less attention due to low performance in productivity which has shifted the interest of the breeders towards temperate cattle breeds to upgrade their local genetic resources. It is generally accepted that the highest amount of genetic diversity in the populations of livestock is found in the developing world where record keeping is poor, and the risk of extinction is high and on the increase. Recently, loss of genetic diversity within indigenous livestock breeds has been a major concern (Kastelic et al., 2005).
Most genetic research is now directed towards the investigation of the relationship between physiological, biochemical and metabolic products/markers to the productive efficiency of farm animals. Biochemical traits, including blood groups, blood proteins and enzymes have been studied with a view to explaining the physiological basis of performance traits.The classical approach to the breeding of superior animals is based on phenotypic variations observed among and between related groups of individuals, but these are only partly due to genetic variations and partly due to environmental influences. There may be however a great variety of genetic variations of a completely different nature that may reflect more accurately the genetic differences and productive efficiency between individuals (Desmarais and Pare, 1974). Polymorphism of blood protein first offered the possibility to study genetic differentiation before the advent of molecular markers. Consequently several livestock breeds including the domestic cattle, sheep and goat have been characterized for variations in major blood proteins (Di Stasio,
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1997). In addition to several important functions of blood proteins, several studies in cattle and sheep have already linked these markers to production traits and environmental adaptation (Vicovan and Rascu, 1989; Charon et al., 1996, Akpa et al., 2010). Blood polymorphism studies have been conducted extensively to identify biodiversity among livestock. Biochemical particles of blood can be determined easily at the post-natal period of young animals, and these components are merely or not affected by the environmental factors. Many research works have been conducted to detect the different types of blood components such as haemoglobin, albumin, glutathione, transferrin and potassium (Kuwar et. al., 2000). Rako et al. (1964) observed that higher productivity may be as result of special traits arising from certain biochemical processes in the organism and suggested the introduction of selection tests based on these biochemical traits. Existence of any significant relationship between activities of enzymes and other biochemical features with performance traits may help to identify a selection criterion which can be used in early life and minimize later recording of performance traits (Singh et al., 1983).
The existence of blood potassium and glutathione polymorphisms in cattle (Evans and Phillipson, 1957; Gonzales et al., 1984), sheep (Gurcan et al., 2010) and goats (Soysal and Ulku, 1998) have been established in some studies. Also several studies were carried out to establish the association between potassium and glutathione polymorphism of blood and various traits (Alpan and Ertugrul, 1991). If an association exists, then the erythrocyte potassium and glutathione types in blood can be utilized as a polymorphic marker among domestic animals (Lush, 1977). Even if some studies did not find any significant relationship with production traits (Soysal, 1983), a few studies displayed an important association between glutathione types and milk production traits in Finn sheep (Atroshi and Soudholn, 1982) and between the potassium
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and sodium types and some economical yield traits in livestock (Antunovic et al., 2004; Milewski and Szczepanski, 2006). Haemoglobin variants have been extensively studied in Zebu cattle and at least eight variants have been identified. Four migration bands were found, Hba, Hbl, Hbc and HbB, but the last band (HbB) may be possibly broken into two, named; HbB1 and HbB2. The respective gene frequencies were 0.563 + 0.012, 0.007 + 0.01, 0.021 + 0.002, 0.188 + 0.007 and 0.221 + 0.007. The genetic frequencies were in equilibrium (Mario, et. al., 1982). The existence of two types of haemoglobin; Hb and HB has been established (Huisman, et al., 1959). They are expressed as homozygous HbAA and HbBB and phenotypes with HbAC being a pre-adult form of Hb (Johnson et. al., 2002). Discrete differences in oxygen affinity of haemoglobin type A and B could also be established in cattle (Huisman, 1959).
1.1 Justification of the Study
Knowledge of the type of biochemical polymorphism and their association with productive and reproductive performances in animals have been successfully utilized in selecting superior performers in recent time (Guney et al., 2003 and Das et al., 2004). However few experiments linked various body features or physical characteristics, biochemical polymorphism and production traits. Some studies are available on some breeds of cattle from East Africa and Brazil on blood biochemical polymorphism, but not much study has been carried out on the breeds of cattle in Nigeria. This therefore necessitated this work.
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1.2 Objectives of the Study
i. To determine blood biochemical activities in relation to morphology and milk traits among three genetic groups of cattle. ii. To determine the blood biochemical polymorphism and their gene and genotypic frequencies among three genetic groups of cattle. iii. To establish the existing relationship among three genetic groups of cattle using Multivariate analysis of Morphometric and milk trait measures.
1.3 Hypotheses of the Study
i. Ho: There is no association in blood biochemical activities in relation to morphology and milk traits among three genetic groups of cattle ii. Ho: There is no association among the blood biochemical polymorphism and their gene and genotypic frequencies among three genetic groups of cattle iii. Ho: There is no relationship between analysis of Morphometric and milk trait measures among three genetic groups of cattle

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