SEMEN QUALITY AND EGG HATCHABILITY IN LOCAL TURKEY FED DIETS CONTAINING MORINGA OLEIFERA AND GONGRONEMA LATIFOLIUM LEAF MEAL

TABLE OF CONTENT

Title                                                                                                                                         i

Certification                                                                                                                ii

Dedication                                                                                                                               iii

Acknowledgement                                                                                                      iv

Table of content                                                                                                v

List of Tables                                                                                                          vi

List of Figures                                                                                                  vii

Abstract                                                                                                                  viii

Chapter One

1.0 Introduction                                                                                                     1

1.1 Background of the study                                                                                    1

1.2 Problem statement                                                                                              3

1.3 Objectives of the study                                                                                        4

1.4 Justification of the study                                                                             5

Chapter Two

2.0.0     Literature Review                                                                                  7                                                                

2.1.0    Origin and Distribution of Turkey                                                               7

2.1.2    Description of Local Turkey                                                                7

2.1.3    Turkey Production in Nigeria                                                        8

2.2.1    Body Weight and sexual maturity                                                   8

2.2.2    Body Weight and Semen Quality                                                         9

2.3.1     Physiology of semen production                                                        9

2.3.2    Physiology of Avian Sperm                                                                       10

2.3.3    Lipid Peroxidation of Semen                                                                      11

2.3.4    Metabolic Aspect of Antioxidant Defense.                                              12

2.4.0    Enhancing Reproductive Efficiency of Turkey                              13

2.5.0    Origin and Distribution of Moringa Oleifera                                     14

2.5.1    Nutritional Properties of Moringa Oleifera                                             15

2.5.2     Amino acid Content of Moringa Oleifera leaves                          15

2.5.3    Therapeutic Properties of Moringa Oleifera                                          16

2.5.4    Effect of Moringa Oleifera on Reproduction in Male                          17

2.6.0    Origin and Distribution of Gongronema latifolium                         17

2.6.1    Nutritional Properties of Gongronema latifolium                                   18

2.6.2   Therapeutic Properties of Gongronema Latifolium                             18

2.6.3   Effects of Gongronema latifolium on Reproduction of Male Animal                           19

2.7.0   Artificial Insemination                                                                        19

2.7.1    Artificial Insemination in Turkey                                                         20

2.7.2    Semen Collection                                                                              21

2.7.3    Semen Quality Evaluation                                                                   21

2.7.3.1  Semen Colour                                                                                    21

2.7.3.2  Volume of ejaculate                                                                       22

2.7.3.3  Motility Evaluation                                                                                 22

2.7.3.4   Motility Evaluation Technique                                                     23

2.7.3.5    Morphology Semen Evaluation                                                  23

2.7.3.6   Morphology Assessments Technique                                        24

2.7.3.7   Sperm Concentration                                                                            25

2.8.0      The Biochemistry of Semen                                                                 26

2.8.1      Determination of Fructose Concentration                                        27

2.8.2       Fructose as a Constituent of Seminal Plasma.                         28

2.8.3       Importance of Fructose Test in Evaluation of Fertility           28

2.8.4       Evaluation of Seminal Chemical Elements on Fertility                  30

2.8.5       Sodium and Potassium Concentration in Semen                   31

2.8.6       Sodium and Potassium Effects on Semen Quality and Fertility              31

2.9.0      Factors affecting poultry semen                                                32

2.9.1      Ambient Temperature                                                              32

2.9.2       Micro Bacterial Contamination                                         32

2.9.3      Photoperiod                                                                                       33

2.9.4      Nutrition                                                                                          33

2.9.5      Age Factor                                                                                        34

2.9.6      Oxidative stress                                                                                     34

2.9.7      Frequency of Ejaculation                                                                    34

2.9.8      Breed/species variation                                                                     35

2.9.1      Semen Collection Technique                                                     35

2.10.1     Artificial insemination                                                                      35

2.10.2     Site, Depths and Time of Insemination                         36

2.10.3     Fertilizing Capacity of the Sperm Cell in vitro                                  37

2.10.4    Duration of Fertile Period in Turkey Hen                                          38

2.10.5     Evaluation of Fertility and Hatchability                                      39

2.11.0    Factors influence Fertility                                                            39

2.11.1  Age Factor                                                                                                 40

2.11.2  Body weight of the Hen                                                                  40

2.11.3  Nutrition                                                                                             41

2.11.4  Stress                                                                                                41

CHAPTER THREE

MATERIALS AND METHODS                                                           42

•   Location  and Duration of the study                                                    42

3.2. Plan of the Study                                                                       42

3.3. EXPERIMENTAL MATERIALS                                                               42

3.3.1. Materials and Processing                                                                        42

3.3.2  Procurement and Management of Experimental Animals                43

3.3.3. Training of Toms for Semen Collection                                                      44

3.4 Data Collection                                                                                              44

3.4.1 The Effect of M. oleifera and G. latifolium on Body Weight:                  44

3.4.2. Semen collection                                                                                45

3.5 Semen Evaluation                                                                                           45

3. 5.1 Semen Colour                                                                                     45

3.5.2 Semen volume                                                                                   45

3.5.3 Motility Evaluation                                                                               46

3.5.4 Sperm Concentration                                                                                 46

3.5.5 Dead and Live /Normal and Abnormal Spermatozoa                              47

3.5.6    Sperm Morphological Evaluation                                                     48

3.5.7    Biochemical Analysis                                                                         48

3.6       Fertility trial (Phase 1V: Fertility and Hatchability)                              48

3.6.1    Artificial insemination                                                                          48

3.6.2    Egg collection, storage and hatchability                                               49

•   Experimental Design                                                                             50
• Statistical analysis                                                                                                       50

Chapter Four

RESULTS AND DISCUSSION

CHAPTER FIVE

5.0. 0  Summary and Recommendation                                                                   78

5.1. 0  Summary                                                                                78

5.2.0   Recommendations                                                                     79

LIST OF TABLES

Table 1:   Vitamin and mineral content of Moringa Oleifera leaf     15

Table 2:  Phytochemicals/Vitamins composition of Gongronema Latifolium                        18

Table3:  Seminal characteristics of Domestic animals                         26

Table 4:   Species Differences in Chemical Composition of Seminal Plasma 30

Table 5: Composition of the Experimental Diets M. oleifera (MO): G. latifolium (GL)        43

Table 6: Treatments combination of Factorial Experimental Arrangement      44

Table7: Treatment Arrangement                                                                 44

Table 8: The Effects M. oleifera on Semen Characteristics of Turkey Toms                           52

Table 9: The Effects of G. latifolium on Semen Characteristics of Turkey Toms                                57

Table 10 Combined Effects of M. oleifera and G. latifolium on Semen quality Traits of Toms.          62

Table 11: Effects of M. oleifera and G. latifolium on Fertility and Hatchability of Turkey Eggs         66

Table 12: Combined Effect of M. oleifera and G. latifolium on Fertility and Egg Hatchability of Turkey Tom’s semen                                70

Table 13: Chemical Composition of Semen 0f Turkey supplemented with M. oleifera or G. latifolium                                                               72

Table 14: Chemical Composition Turkey Semen Fed Combined level of M. oleifera andG. latifolium                                                                74

Table 15:   Measures of Association Between Body Weight and Semen Characteristics of Tom                                                                                76

LIST OF FIGURES

Figure 1: Semen collection, evaluation and insemination                       45

Figure 2: Slide preparation                                                               48

Figure 3: Effect of treatments on body weights of Turkeys across weeks                             51

Figure 4: Viability and morphological examination (Stained spermatozoa)                           56

Figure 5:  Hatched live poult and dead in-shell embryos              71

ABSTRACT

The experiment was conducted to determine semen quality, fertility, egg hatchability and some biochemical parameters in Nigerian local turkey toms fed diets containing Moringa oleifera (MO), Gongronema latifolium (GL) leaf meals and their combinations. A total of 72 Nigerian local turkeys comprising of 54 males and 18 females were used for the study. The males were randomly divided into 9 treatment groups, each treatment was replicated 3 times with 2 toms per replicate. The treatment diets were given only to the toms, starting from three month of age through the experimental period. The experimental animals were fed and given water properly, twice a day without restriction. All the management practices were carried out to the best of ability. The males in all the treatment groups were weighed weekly to determine their daily and weekly body weight gain. At 26 weeks of age, toms were trained for semen collection, and 32 weeks of age, semen was collected using abdominal massages. Samples were analyzed for colour, volume, progressive motility, sperm concentration, viability and sperm morphology. Fresh semen sample were also collected per treatment in vials’ stored in ice block and analyzed for fructose, Na and K.  A total of 18 hens were randomly shared 2 per treatment corresponding to the 9 treatments. Pooled Semen from each treatment was used to inseminate the hens twice a week at the beginning of egg lay and once a week subsequently. A total of 225 eggs were collected and incubated in weekly batches, analyzed for fertility and hatchability. The result revealed that M. oleifera and G. latifolium leaf meals had significant (P<0.05) effects on the semen quality parameters measured. M. oleifera fed tom at 3kg yielded the best result: ejaculate volume 0.58ml, motility= 92.93%, Conc.= 4.82(x10/ml ⁹), live sperm= 94.13%, normal sperm 91.38% and corresponding lower values for percentage dead and abnormal sperm. While, G. latifolium treated toms had a lower value for their semen quality parameters when compared with the control group. Interaction effects of M. oleifera and G. latifolium leaf meals were significantly (P<0.05) different. Compared with control semen quality traits were higher at 3kgMO+1.5kgGL inclusion, lower at 1.5kgMO + 1.5kgGL, and significantly reduced semen quality of toms fed 1.5kgMO+ 3kgGL diets.  Similarly, the percent fertile eggs, and percent hatched eggs were greatly improved at 3kgMO diets with corresponding decrease in percent infertile eggs and dead in shell embryos than the control. However, toms fed diet at 1.5kgGL and 3kgGL had their values for these parameters being severely reduced. Hatchability values increased to 88.39%  and 83.33% at 3kg MO+1.5kgGL and 3kg MO+3kgGL respectively with a decrease in percent dead- in- shell embryos to 16.99% and 19.12% respectively. Seminal fructose concentration (mg/100ml) was significantly (P<0.05) increased (5.86+2.76) at 3kgGL when compared with the control, but M. oleifera had a negligible increase in fructose concentration. However, M. oleifera fed toms (3kg) had a significant(P<0.05) increase in concentration of Na and K (0.39 and  0.35) respectively. These result suggest that improved fertility, eggs hatchability and reduction in percent embryo mortality can be achieved using M. oleifera at 3kg/100kg diet  and combination of M. oleifera+G. latifolium at rate of 3kgMO+1.5kgGL, but treatment with G.latifolium at the rate of 1.5kg, 3kg and combination at rate of 1.5kgMO +3kgGL caused reduced fertility in local Nigerian turkey.

CHAPTER ONE

INTRODUCTION

1. Background of the study

In Nigeria, poultry industry is once again experiencing growth due to the current regime’s effort at encouraging investments in the industry through several economic and agricultural policies and reforms including removal of import duties on agricultural products (Fasina et al., 2007). Notwithstanding, the current trend in growth within the industry it is still experiencing challenges as some species of poultry are left out. For instance, turkey production has not been as successful as chicken production in Nigeria. Its production is largely at the small holder level. This has been attributed to high cost of feed, inconsistencies in feeding program as well as lack of information on its nutritional requirements (Ojewola et al., 2002). Also, reproductive problems experienced under natural mating conditions, low fertility and poor hatchability as a result of poor quality semen due to oxidative stress amongst other factors (Bucak et al., 2010) militate against turkey production in Nigeria.  This situation is also evident from the FAO report (FAOStat, 2011), which shows that the population of  local turkeys in Nigeria is only about 1.05 million, being the smallest when compared with other poultry species. It is important to come to terms with the fact that advancement in the industry depends on the use of birds with high reproductive rate, adoption of better mating methods, use of high quality semen in insemination as well as good nutrition.

According to Donoghue and Donoghue (1997), avian spermatozoa are rich in polyunsaturated fatty acids (PUFA) which makes them vulnerable to lipid peroxidation especially during in-vitro manipulation. In particular fatty acids are the most vulnerable to lipid peroxidation. Generally, some features of avian semen have also been found to put it under pressure of oxidative stress. For instance, there is limitation in antioxidant recycling, because of very low activity or even absences of hexose mono-phosphate shunt in avian spermatozoa (Sexton, 1974). Also, the low production of NADPH (the coemzymes for glutathione reductase) has been implicated as a factor in reducing fertility of avian sperm. There are also observations that leukocyte contamination of the semen is responsible for increased generation of free radicals which affect the performance of turkey sperm (Halliwed and Gutteridge, 1999). Furthermore, the activity of antioxidant enzymes in turkey spermatozoa is also lower compared to that of chicken and this makes turkey sperm more vulnerable to the problem of peroxidation (Aitken, 1999). Worthy of note, is the fact that turkey spermatozoa are very dependent on oxidative metabolism to maintain optimal ATP level needed for sperm metabolism (Wishart, 1982). Therefore, any damage resulting from these discrepancies may leads to alteration of the membrane irreversibly, thereby affecting sperm function and fertilizing ability. In effect, antioxidant protection is thus absolutely vital for maintaining the fertility of turkey spermatozoa.

Studies have revealed how the reproductive efficiency of male breeder can be affected by a variety of factors such as breeding methods, environment (daily photoperiod, temperature housing, and nutrition) and frequency of semen collection and technique of artificial insemination (AI), especially in turkey (Sexton, 1983 and Lake, 1984). In addition, these authors have stressed the importance of evaluating the semen prior to insemination to improve the reproductive efficiency. Antioxidants have been reported to be efficient in diminishing lipid oxidation in avian spermatozoa which is a major factor in reduction of fertility. Worthy of note is the fact that natural antioxidant has the ability to increase the antioxidant capacity of the seminal plasma and reduce the risk of certain deleterious free radicals on sperm fertilizing ability (Chanda and Dave, 2009). Dawson et al. (1990) reported that the antioxidant properties of ascorbic acid are essential in maintaining the membrane and the genetic integrity of sperm cells by preventing oxidative damage to the sperm DNA. Also, studies have shown that antioxidants especially those of plant origin such as Moringa oleifera and Gogronema Latifolium have greater application potential for therapeutic and reproductive uses.

Moringa Oleifera plant in the family of Moringacea is native to India, naturalized in tropic and sub-tropical areas of the world (Price, 2002). It is widely distributed and cultivated in the northern part of Nigeria and it is called Zogale in Hausa. The plant is characterized as fast growing and drought resistant with an average height of 12 meters at maturity (Fuglie, 2001). All parts of the moringa tree is said to have beneficial properties. Nutritional analyses by Gopalan et al. (1989) and Fuglie (2001) indicate that Moringa leaves contain a wealth of essential amino acids, vitamins and minerals with higher values in their dried form than in its fresh form, except for vitamin C which is high in its fresh leaves. Fuglie (1999) also reported some specific plant pigments with demonstrated anti oxidant properties such as carotinoids, lutein, alpha-carotine, beta-carotine, xanthins and chlorophyll. Other phytochemicals contained in moringa which have powerful antioxidant ability include kaempferol, queretin, rutin, kaffeoylquinic acids, vitamins A, C and E, some valuable micro nutrients such as selenium and zinc are also found in the leaves of Moringa.

Recently, scientists have put more interest on the role of this plant in improving male reproduction efficiency. Cabacungan (2008) reported that a steady diet of moringa fruit boost the sperm count of male thus, improving their chances of fertilizing an egg. Interestingly, Serrano M. R (2008) reported an increase in sperm count in male mice when 1% concentration of moringa ethanolic leaf extract was administered. Cajuday and Pocsidio, (2010) also observed that mice administered with high and medium dose of the plant extract had enhanced spermatogenesis. This evidence was supported by increase in testicular and epididymal weights as was confirmed in the report of Gonzales (2001).

On the other hand, Gongronema latifolium of the family asclepiadaceae is a tropical rainforest plant primarily used as spice and vegetable and in traditional folk medicine. It is commonly called Utazi by the Igbo tribe in South Eastern of Nigeria. (Ugochuku et al., 2003; Ugochuku and Babady, 2002).  Phytochemical screening of the ethanolic extract of the plant shows that the root contains poly-phenol in abundance, Alkanoids, glycosides and reducing sugars in moderate amounts (Antai et al., 2009). Other chemicals such as B-sistosterol, lupenylester, pregnane ester and essential oil were found in the plant extracts as reported by Ekundayo (1980). Atawodi, (2005) also reported the antioxidant potentials of the plant, which was confirmed by the report of Nwanjo et al. (2006). In addition, the plant is suggested to be able to mop up reactive oxygen species in the system. According to Ugochuku and Babady (2002); and Ogundipe et al. (2003) ethanolic and aqueous extracts of the plant had hypoglycemic, hypolipidermic and antioxidant properties.

Evaluation of biochemical constituents of semen is an important criterion for assessing male fertility. Biochemical constituents of seminal plasma are said to play a role as sperm metabolites, nutrition of ejaculated sperm and provision of protection to spermatozoa against proteinase inhibitors, which help in sperm capacitation and local immunosuppression (Pesch et al., 2005). Therefore, ensuring that the various major biochemical constituents of semen are available in there right proportions is an indication of semen quality.

1.2 Problem Statement