ABSTRACT
Effect of garlic and vitamin E supplementation on gonadal and extra-gonadal sperm characteristics, hematological indices, serum biochemistry, linear body measurement, reproductive hormone and testicular histology of pre pubertal buck rabbit were investigated. A total of 36 bucks with average weight of 1.76kg were used for the 16 weeks experiment. The bucks were randomly assigned to four dietary supplementation and three replicates in a completely randomized design (CRD). They were 9 rabbits per treatment with 3 rabbits per replicate. One dietary treatment was formulated to meet the nutrient requirements of rabbit at four supplementation rate of 0% garlic and vitamin E (T1), 3% garlic (T2) vitamin E (T3), and 3% garlic and vitamin E (T4) respectively. Inclusion of vitamin E was at the rate of 3000ɩµ per kilo gram of feed. Performance parameter such as linear body measurement was taken at end of the experiment. Hematological indices, serum biochemistry, organ weight, semen characteristic, reproductive hormone, Testicular morphology and histology were all examined at the end of the experiment. Data collected on different parameters were subjected to analysis of variance (ANOVA). Mean procedure and ANOVA procedure was used to generate the mean and standard error of mean. Significant means were separated using Duncan Multiple range test. Results showed significant increase (P<0.05) in final body weight, ear length, fore limb, hind-limb and body length with range of values; 2.140kg-2.383kg, 11.665cm-13.250cm, 15.165cm-16.882cm, 26.000cm-27.832cm, 11.332cm-12.332cm, 5.082cm-6.165cm and 33.082cm-38.082cm respectively for rabbits fed diet T2 and T4. There was significant increase in tail length for bucks on T4 while T4 was similar with the control on the head to shoulder. Haematological parameters showed significant increase in red blood cell (RBC) count for vitamin E group (T3), group 2 (T2) was higher on mean corpuscular haemoglobin concentration MCHC while White Blood Cell (WBC) count decreased with supplementation. T4 monocytes value was significantly (P<0.05) higher. Serum biochemistry characteristics showed significantly higher value on total protein, globulin and albumin for group 3 (T3). The serum bilirubin and serum enzyme values proved T2 higher on creatinine and glucose value. Serum enzymes values showed significantly (P<0.05) higher ALT in T3 and decreased ALP with supplementation. Significantly (P<0.05) higher result were observed in all the parameter in semen evaluation except the semen volume which were similar in all the treatments with values ranging from 0.53ml-0.60ml. Testicular weight was increased with supplementation. Testicular weight was higher on T4. Reproductive hormone, Follicle stimulating hormone and testosterone increase with supplementation while the luteinizing hormone decreased. Testicular histology showed increased in density in interstitial space of the seminiferous tubules with supplementation. The result proves that both garlic and vitamin E improve semen production but combination of the two gives a better result. However garlic and vitamin E should be used together for quantitative and qualitative semen production.
TABLE
OF CONTENTS
PAGE
Title
Page i
Certification ii
Declaration iii
Dedication
iv
Acknowledgement v
Table
of Contents vi
List
of Tables xi
List
of Plates xii
Abstract xiii
CHAPTER 1: INTRODUCTION 1
1.1 Statement of
Problem 4
1.2 Justification 5
1.3 Objective of the Study 5
CHAPTER 2: LITERATURE
REVIEW
2.1 History of Rabbits 6
2.2 Rabbit Production 6
2.3 Breeds of Rabbits 7
2.4 Chinchilla 7
2.5 New Zealand White 7
2.6 System of Management 7
2.7 Handling 7
2.8 Housing 8
2.9 Feeding 8
2.10 Uses of Rabbit 9
2.11 Anatomy of Rabbits 9
2.12 Musculoskeletal System 9
2.13 Dentition and Oral Cavity 10
2.14 Skin 10
2.15 Eyes and Ears 11
2.16 Digestive Tract 12
2.17 Respiratory System 14
2.18 Cardiovascular System 14
2.19
Urinary System 14
2.20 Physiology of Male Reproductive System 15
2.21 Influence Garlic and Vitamin E on
Testosterone 16
2.22 Spermatogenesis 16
2.23 Libido 16
2.24 Semen 17
2.25 Semen Quality 17
2.26 Semen Characteristics 19
2.27
Semen Colour 19
2.28 Semen Volume 19
2.29 Sperm Concentration 20
2.30 Live Sperm Percentage 20
2.31 Sperm Motility 20
2.32 Semen Ph 21
2.33 Acrosomal Morphology 21
2.34 Abnormal Spermatozoa 21
2.35 Role of Seminal Granules in the Physiology
of Sperm After ejaculation 22
2.36 Artificial
Insemination 22
2.37
Artificial Vagina (AV) 24
2.38 Haematology 24
2.39 Haematological indices 25
2.40
Leucocytes 25
2.41 Erythrocyte Count 25
2.42 Serum Metabolites 25
2.43 Serum Enzymes 26
2.44 Serum Electrolytes 39
2.45 Linear Body Measurement 29
2.46 Spermatozoa,
Synthetic and Natural Fertility Drugs 30
2.47 Spermatozoa and
Antioxidant (Tocopherol) 31
2.48 Historical Perspective of Garlic 32
2.49 Garlic Preparations and their Chemical
Compounds 33
2.50 Atherosclerosis and Lipid Metabolism 34
2.51 Factors Affecting the Use of Garlic 35
2.52 Fibrinolytic Activity 37
2.53 Platelet Aggregation 38
2.54 Blood Pressure Lowering Effect 42
2.55 Diabetes Mellitus 44
2.56 Cardioprotective Properties of Garlic 44
2.57 Hormonal
Activities of Garlic (Reproductive Hormone) 46
2.58 Adverse Effects 47
CHAPTER 3: MATERIALS AND
METHODS
3.1 Experimental Location 50
3.2 Materials and Methods 50
3.3 Experimental Animals (New Zealand White)
and Management 50
3.4 Experimental Diet 51
3.5 Sources and Processing of Garlic 51
3.6 Experimental Design 52
3.7 Data Collection 53
3.8 Libido (Reaction Time) 53
3.9 Semen Collection 53
3.10 Semen Evaluation 54
3.11 Semen Volume 55
3.12 Sperm Motility 55
3.13 Live Sperm Percentage 55
3.14 Sperm Concentration 56
3.15 Morphology 56
3.16 Morphometric Measurement 56
3.17 Scrotal Morphometry 56
3.18 Testicular Morphometry 56
3.19 Testicular Histomorphometry 57
3.20 Linear Body Measurement 57
3.21 Organ Weight Characteristics 58
3.22 Blood Collection 58
3.23 Analysis of Blood Samples 59
3.24 Haemoglobin Concentration 59
3.25 Erythrocyte Count 59
3.26 Packed Cell Volume (PCV) 59
3.27 Leucocytes Count 59
3.28 Blood Indices 59
3.29 Biochemical Analysis 60
3.30 Hormonal
Assay Procedures 60
3.31 Data Analysis 61
CHAPTER 4: RESULTS AND
DISCUSSION 62
CHAPTER 5: CONCLUSION
AND RECOMMENDATIONS 89
References 90
LIST OF TABLES
1: Mean and Normal Range of Haematological Values of Rabbits 27
2: Mean and Normal Range of Biochemical Values of Rabbits 28
3: Performance Characteristics of Rabbits
Under Tropical Condition 30
4: Weekly Body Weight, Heart Girth, Length at Withers and Body
Length
of Rabbits 30
5: Gross Composition
and Calculated Nutrients of Experimental Diets
for Pre-pubertal Rabbit Bucks 52
6: Scores
and Description for Levels of Libido 53
7: Scores
and Description for Progressive Motility 55
8: Linear
Body Measurements of Rabbit Bucks Fed Diets
Supplemented with Garlic and Vitamin E. 62
9: Haematological Characteristics of Rabbit Bucks Fed Diets
Supplemented with Garlic and Vitamin E. 64
10: Serum
Biochemistry Characteristics of Rabbit
Bucks Fed Diets
Supplemented with Garlic and Vitamin E. 68
11: Serum
Bilirubin and Serum Enzyme Values of Buck Rabbits Fed
diet
Supplemented with Garlic and Vitamin E 71
12: Organ Weight Characteristics
of Rabbit Bucks Fed Diet
Supplemented with Garlic and Vitamin E 73
13: Semen
Quality Characteristics Buck Rabbits Fed Diet Supplemented
with
Garlic and Vitamin E. 76
14: Seminal Plasma Characteristic of Buck Fed Diet Supplemented
Garlic
and Vitamin E 80
15: Testicular
Morphomemetric of Buck Rabbits Fed Diet
Supplemented with Garlic and Vitamin E 81
16: Male Reproductive Hormone of Bucks Fed Diets Supplemented
Garlic and Vitamin E 84
LIST OF
PLATES
1a: Section
of Control Testis Showing Normal Architecture of the
Testis H&E X100 85
1b: Section of Control
Testis Showing the Lumen of the Seminiferous
Tubules H&E X400 85
2a: Section of T2
Testes on Garlic Supplementation.
H&E×100 86
2b: Section of T2
Testis Showing the Lumen of the Seminiferous
Tubules
H&E X400 86
3a: Section of T3
Testis on Vitamin E Supplementation.
H&E×100 87
3b: Section of T3
Testis Showing the Lumen of the Seminiferous
Tubules.
H&E X400 87
4a: Section of T4
Testis on Garlic & Lumen of the Seminiferous
Tubules. H&E X400 88
4b: Section of T4 Testis Showing the Vitamin E
Supplementation. 88
CHAPTER 1
1.0 INTRODUCTION
In the face of the Rising need for animal protein supply to
satisfy the ever teaming human population in the developing countries, coupled
with the prohibitive costs of the conventional feed stuffs, it has become
imperative to explore rabbit production potential. Animal protein consumption
in Nigeria and other developing countries has been reported to be comparatively
low (Ayorinde and Aromolaran, 1998). This had been traced to insufficient
supply of conventional protein product of animal origin, drop in the
reproductive capacity of most animal, resulting in a high cost of most animal
products essential for human nutrition. However animal protein can be obtained
from micro-livestock such as rabbit, grass cutter and snail, which also are high
in protein, (Betz and Wiedler, 1989).
According to (Mamattah, 1978) rabbit have no apparent competition
with humans for food and they are socially accepted on the combined merits;
their low space requirement, high reproductive rate, minimal zoonotic health
hazard, minimal capital investment as well as absence of social taboos against
the consumption of their meat. Rabbit is a good source of quality meat with a
balanced amino acid profile. The rabbit meat is highly digestible, wholesome,
tasty, low in cholesterol, sodium and fat but high in protein, (Biobaku and
Oguutona, 1997). Ajaji et al. (2012)
reported that rabbit has a number of characteristics such as small body size,
short generation interval which is as a result of short gestation period of
30-31 days, high reproductive potential, rapid growth and ability to utilize
forage and fibrous agricultural by-products. The author maintained that these
attributes make rabbits sexually mature animals 30% faster than other animals.
In spite of all these
advantages over other livestock, rabbit production has not achieved its
potential as cheap animal protein source in the tropics (Herbert and Adejumo,
1995).
Reproductive inefficiency
is the most constraint to efficient rabbit production in the tropics (Murray
and Meacham, 1993). The efficiency of sperm production, libido and quality of
sperm tend to remain uniform through out the reproductive life of an animal but
may be significantly altered by age, nutrition, environment, health status,
drugs, and chemicals (Togun and Egbunike, 2006).
Among these factors
nutrition, drugs and hormones are the most prominent. Sexual maturity is known
to be delayed by a poor nutrition regiment during growth (Omole, 1982). It also
affects age at puberty and stimulation of hypothalamus indirectly to produce
interstitial cells stimulating hormone that acts in the testicular tissue (Cogan
et al. 2004).
Vitamin E (tocopherol) is the major scavenger
for reactive oxygen species ROS inside membranes of body tissue (Welers and
Sies, 1988). It has been reported that vitamin E inhibits lipid peroxidation by
scavenging lipid peroxyl radicals much faster than the radicals can react with
adjacent fatty acid side chains or membrane proteins (Machlin, 1984; Welers and
Sies, 1988). Mmoa et al.(2012) reported
that supplementation with vitamin E reduced the generation of reactive oxygen
metabolism ROMs and prevented loss of motility, mainly during the storage of
spermatozoa. According to (Castellini et
al. 2000), restricted dietary protocol reduces libido and some sperm
quality trait.
Alium sativum commonly known as garlic is a vegetable species that can be
classified as either a food or a medicinal herb. It is a versatile vegetable
often used as ingredient in many dishes for perma, aroma and taste enhancement,
(Nasibeh and Arash, 2014). It is a
good source of dietary phytochemicals with proven antioxidant properties and
ability to modulate the detoxification system, (Elkayam et al. 2003).
The important components among others include allicin (allyl
2-proenethiosulfinate or diallythiosulfinate), which is thought to be the chief
bioactive compound present in aqueous garlic allyl methyl thisulfonate,
1-propenyl allylthiosulfonate. When crushed, Allium sativum yield allici, an antibiotic (Focke et al., 1990) and antifugal compound
(phytoncide) discovered by Cavallito and colleagues in 1944. Fresh or crushed
garlic also has enzymes, B vitamins, proteins, minerals, saponins, flavonoids
and maillard reaction products. Futher more, a phytoalexin (allixin) was found,
a nonsulfur compound with a y-pyrone skeleton structure with antioxidant
effects, antimicrobial effect (Aiten and Fisher 1994) antitumor promotive
effects (Aaron et al. 2001),
inhibition of aflatoxin B2DNA binding and neurotrophic effects (Aaron et al. 2001). Although the active
ingredient in garlic is said to be unknown, there is a lot of speculation
around allicinas being the main constitutive candidate (Hammami and El-May
2012).
Allicin is a very unstable molecule and break down very easily.
This is why it is recommended to eat fresh garlic as it contains highest amount
of allicin. It is also the reason for strong odour that garlic possesses. Due
to this instability, medical companies have now started to develop garlic
supplements that contain alliina stable precursor to allicin.
The effects of garlic on both plant and animal models lipids is
best known for its ability to decrease the amount of lipids or free fatty acids
as well as its anti-atherogenic effects in both models (Prasad, 2010). Allium
products have the potential to reduce the risk of developing cancer or
potential to decrease risk factors associated with cancers. However, feeding higher
garlic supplementation over a longer period of time to adult rat causes an
increase in epididymal spermatozoa, (Hammami and El-May 2012).Meanwhile,
studies by (Oi et al. 2001) found an
increased testicular testosterone with garlic supplementation. This study
focuses on evaluation of the effects of Allium
stativum and vitamin E supplementation on organ development, male
reproductive hormone and sperm characteristics of rabbit buck.
1.1 STATEMENT OF PROBLEM
The human population in developed countries is stabilized while
that of developing countries including Nigerian is still increasing rapidly, (Massanyi
et al., 2000). Thus the search for
alternative sources of protein to meet up the population challenges is
imperative. Economic indices indicate that as this population trend continues
more people are to be fed and demands that agricultural output needs to be
increased rather than through food importation into such countries (Allen and
tresini, 2000). In order to maximize food production and meet protein
requirement in Nigeria viable options need to be explored and evaluated (Owen,1992). Among such alternatives is the
use of livestock species that are yet to play a major role in animal production
in the country. Fast growing livestock such as rabbits possess a number of
features that might be of advantage in the small holder subsistence type
integrated farming in Nigeria.To achieve these, effort should be geared toward
enhancing the effectiveness of the reproductive performance of rabbit buck
through improvement of semen and the internal organ development. Although many
positive health effects have been attributed to the use of garlic, fighting
against microbes existing in respiratory system, increase testicular
testosterone and reduces other internal organ diseases (Hammami et al. 2009). The effects tend to be
undesirable on testicular function as many studies have observed, some adverse
effects on reproductive functions such as inhibition of leydigsteroidogenic
enzyme and sertolicell markers which are capable of inducing apoptosis in
testicular germ cells (spermatocytes and spermatids) characterized by increased
levels of active caspase (Banerjee et
al., 2001 and Khaki et al.,
2013).
1.2 OBJECTIVE OF THE
STUDY
Arising from this background, this study
is designed with the general objective of determining the gonadal and
extra-gonadal sperm characteristics and organ development of rabbit bucks with
dietary supplementation of garlic and vitamin E.
The specific objectives were to:
1.
Determine the
effect of garlic and vitamin E supplementation on the gonadal and extra gonadal
sperm characteristics of rabbit bucks.
2.
Evaluate the
effect of garlic and vitamin E on development of internal organs of rabbit
bucks such as; liver, kidney, lungs etc.
3.
Assess the
effect of garlic and vitamin E supplementation on male reproductive hormone of
rabbit bucks
4.
Examine the
effect of garlic and vitamin E supplementation on testicular histomorphology of
rabbit.
5.
Evaluate the
effect of garlic and vitamin E supplementation on haematological/serum
biochemistry of rabbit bucks
1.3 JUSTIFICATION
The retardation of male reproductive
function is one of the major manifestations of organ infection and toxicity.
These have prompted the need for exploring the productive effects of some
antioxidants (garlic and vitamin E) on testicular function hormone and internal
organ development of rabbit buck. Although the beneficial effects of garlic and
vitamin E have been exploited in different aspect of animal production, little
has been conducted on its activity on testicular function/semen characteristics
hormone and internal organ development of rabbit buck. Therefore, there is a
need to determine the effects of garlic on semen characteristics, reproductive
hormone and internal organs such as liver, kidney, lung etc of rabbit.
The needed improvement on the semen
characteristics hormone and internal organs of rabbit may be achieved by
investigating the effects of vitamin E and a medicinal plant such as garlic.
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