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- inshell
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.
TABLE OF CONTENTS
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
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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
3.1. 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
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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
3.9 Experimental Design 50
3.9.1 Statistical analysis 50
Chapter Four
RESULTS AND DISCUSSION
4.1 Effects of M. oleifera and G. latifolium on Body Weight (kg) 51
4.1.1 Effects of M. oleifera supplementation on Semen Colour and Volume 52
4.1.2 Effects of Moringa oleifera on Progressive Motility 53
4.1.3 Effects of Moringa oleifera on Sperm concentration 54
4.1.4 Effects of Moringa oleifera on Sperm Viability (Live/Dead) 55
4.1.5 Effects of Moringa oleifera on Sperm Morphology 56
4.2.1 Effects of Gongronema latifolium on Semen Colour and Volume 57
4.2.2 Effects of Gongronema latifolium on Progressive Motility 58
4.2.3 Effects of Gongronema latifolium on Sperm Concentration 59
4.2.4 Effects of Gongronema latifolium on Sperm Viability (Live/Dead Ratio) 59
4.3.1 Effects of Gongronema latifolium on Sperm Morphology 60
4.3.2 Combined Effects of M. oleifera and G. latifolium on Semen colour and Volume 62
4.3.3 Combined Effects of M. oleifera and G. latifolium on Progressive Motility 62
4.3.3 Combined Effects of M. oleifera and G. latifolium on Sperm Concentration 63
4.3.4 Combined Effects of M. oleifera and G. latifolium on Sperm Viability 64
4.4.1 Combined Effects of M. oleifera and G. latifolium on Sperm Morphology 65
4.4.2 Effects of M. oleifera and G. latifolium inclusion on Percent Fertility of toms Semen 66
4.4.3 Effects of M. oleifera and G. latifolium on Percent Dead -in- Shell Embryos 68
4.5.1 Effects of M. oleifera and G. latifolium on Percentage Hatched Eggs 69
4.5.2 Combined Effects of M. oleifera and G. latifolium on Percent Dead-in -Shell Embryos 70
4.5.3 Combined Effects of M. oleifera and G. latifolium on Percent Egg Hatchability 72
4.6.1 Effect of M. oleifera and G. latifolium on Fructose Concentration in Toms Semen 73
4.6.2 Cations Concentration in Tom Semen fed varying levels of M. oleifera and G.latifolium 74
4.7.1 Effects of M. oleifera and G. latifolium on Fructose Composition of Turkey Toms
Semen
75
4.8.1 Associations between semen quality parameters and body weight of treated tom 76
CHAPTER FIVE
5.0. 0 Summary and Recommendation 78
5.1. 0 Summary 78
5.2.0 Recommendations 79
CHAPTER ONE
INTRODUCTION
1.1. 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 invitro
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
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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.
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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
Turkey is one species of poultry that are bred exclusively by artificial insemination due to the
differences in body weight between the male and female. Most times the male weighs twice
more than the female, consequently the larger body sizes (weight) of the male accounts for
poor mating ability. Thus tom often spend more time preparing to mate with the female,
hence the female might lose interest or is weakened due to the long time spent by the male
and this results in ineffective mating. Secondly, studies have revealed that turkey toms are
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naturally clumsy during natural mating and as such when semen is released little or nothing is
discharged into the vulva of the hen turkey as a result of the tom’s awkward mating skills and
thus much of the semen is wasted. Unfortunately, the full potentials of artificial insemination
technique in Nigeria have not been fully utilized and improvement in turkey production is
limited by fewer numbers of experts in artificial insemination technique, particularly those
working on local turkey production.
Furthermore, avian semen has been found to be rich in PUFA which put them under pressure
of oxidative stress (Donoghue and Donoghue, 1997). The activity of antioxidant enzymes in
turkey spermatozoa is lower compared to that of chicken and this makes turkey sperm more
vulnerable to peroxidation (Aitken et al., 1999). In effect, an unsatisfactory egg fertility and
hatchability due to the low quality semen of turkey constitutes a major problem for turkey
breeders. Also, during hatching and early postnatal period, the accumulation of PUFAproducing
tissue makes it vulnerable to peroxidation. In poultry, vitamin E (combine with
selenium) provides protection against lipid peroxidation especially in turkey semen, this
vitamins is effectively transferred both from the paternal and maternal system to the egg yolk
and further to the developing embryo (Surai, 2002).
The biochemical constituents such as fructose, potassium and sodium of semen are important
for sperm fertility and deviation from normal values or proportion of these biochemical
components in seminal plasma may result to low or male infertility (Cevik et al., 2007).
Handler and Bulos(1965) reported that fructose serves as fuel supply for sperm cells, and in
the absence of fructose infertility would occur. In general, deviation from normal values of
biochemical components in seminal plasma is correlated with male infertility (Cevik et al.,
2007). The present study was designed to determine the effects of M. oleifera and G.
latifolium leaf meals in turkey toms’ diets on semen quality, biochemical parameters and egg
hatchability of local turkeys.
1.3 Objectives of the Study
The main objective of the study is to determine Semen Quality and Egg Hatchability in
Local Turkey Fed Diets Containing Moringa Oleifera and Gongronema Latifolium Leaf
Meals.
Specifically this study seeks to:
i. Determine the effects of Moringa oleifera and Gongronema latifolium on daily body
weight of turkey toms.
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ii. Evaluate the effects of Moringa oleifera and Gongronema latifolium on semen quality
parameters.
iii. Determine the fertilizing capacity of the spermatozoa obtained from the treated toms
on hatchability of eggs laid by inseminated hens.
iv. Determine the concentration of some biochemical constituents of turkey semen and
correlate them with some notable semen quality parameters.
v. Correlate body weight of the treated toms with their semen quality parameters.
1.4 Justification
The greatest achievement of every producer in the poultry industry is to maintain breeder
males capable of producing viable spermatozoa that can fertilize eggs which will hatch with
minimum mortality. However, the numbers of hatched eggs is dependent on the quality and
quantity of the spermatozoa, thereby determining the profitability of the production.
Unfortunately, reproductive efficiency in turkey has been compromised due to emphasis on
high body weight during genetic improvement. Artificial insemination therefore, has become
the most effective and widely used techniques in turkey production and for causing
reproductive improvement in most farm animals. The success of artificial insemination is
directly dependent on the quality of semen output and appropriate handling procedures to
sustain the fertilizing potential of the spermatozoa. In Nigeria, the turkey industry has not yet
utilized the high reproductive potentials offered by artificial insemination, as a major tool to
improve and optimize the genetic potential of the local breeds and eliminate the reproductive
challenges in turkey production.
The practice of using drugs or hormones to enhance reproductive efficiency in poultry has
been questioned in many areas because of their cumulative negative health effects in the
animal as well as their products (meat and egg) meant for human consumption. Alternative
measures are now being recommended for improving reproduction in farm animals through
the application of organic extracts of plant components (leaf, seeds, stem and root)
administered through feed or water. Most recently, some available underutilized plants have
been proved to have nutritional, medicinal and therapeutic properties which can improve
semen quality, fertility and even hatchability of the turkey eggs. Moringa oleifera and
Gongronema latifolium contain nutrients, vitamins, minerals, some beneficial phytochemicals
and antioxidants which are known to stimulate growth and improve reproductive efficiency in
humans and animals. The plants have been used as natural feed additives and have generally
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been proven to be effective and non-toxic when consumed by humans and animals. It is
against this background that this research was conducted.
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