CONCEPTION RATES OF ESTRUS-SYNCHRONIZED INDIGENOUS UGANDAN GOATS BY CERVICAL ARTIFICIAL INSEMINATION

BY

Nadiope Gideon (BVM)

A DISSERTATION SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF THE DEGREE OF MASTER OF SCIENCE IN LIVESTOCK DEVELOPMENT PLANNING AND MANAGEMENT OF MAKERERE UNIVERSITY

2010

DECLARATION
I, Nadiope Gideon, do declare that to the best of my knowledge, this work is original and has never been submitted to this or any other university for any award. Signed.Date

This work was conducted under the supervision and approval of: 1. Dr. David Owiny (BVM, MSc, PhD) Department of Veterinary Surgery and Reproduction Faculty of Veterinary Medicine Makerere University SignedDate

2.

Dr. Maria Gorretti Nassuna-Musoke (BVM, MSc, PhD) Department of Veterinary Surgery and Reproduction Faculty of Veterinary Medicine Makerere University

Signed..Date..

DEDICATION
This work is dedicated to the Active Poor and Vulnerable Persons who struggle to improve their livelihood through goat production.

ii

ACKNOWLEDGEMENTS
I am greatly indebted to my family, most especially my lovely wife Mrs. Monica Nadiope for allowing me use part of our family income to finance my studies for this Masters program at Makerere University and for the great encouragement and support. My sincere thanks and gratitude go to my supervisors, Dr. David Owiny and Dr. Maria Gorretti NassunaMusoke, Senior Lecturers of Theriogenology, Makerere University, for their keen personal interests, inspiring criticism and relentless guidance which turned this work into a reality. I do thank all the lecturers at Makerere University, most especially those who facilitated in the Masters of Science in Livestock Development, Planning and Management Program. My heartfelt appreciation goes to my mentor Dr. Lorna Brown of Dolen Ffermio in Wales UK. With her backing, support, encouragement, provision of equipment and materials, and training when I started my experimentation in goat artificial insemination. I sincerely thank Dr. Pradip Ghalsasi of Nimbkar Agricultural Research Institute, Phaltan, India for coming over to Uganda and providing me practical tutorials which have helped me to improve my knowledge, skills and competence in goat artificial insemination. Thanks go to Dr. Johan Steyr and Dr. Farnnie Steyr of South Africa for the part played in my goat artificial insemination training. I am grateful to James and Isabel Birkett who provided the laptop which I used all through this course, Professor Eymr Owen, Dr. Tim Smith and Dr. Denis Mugizi with whom I held consultations and Peter Isabirye with whom we worked on the statistics. My appreciations go to fellow colleagues with whom I have constructively interacted throughout the period of this course. Thanks go to all the research assistants most especially Vincent Bbosa and Richard Mugwanya, all farmers who allowed me to use their goats and assisted me in data collection. I remember my parent, sisters, brothers, friends and all those who assisted me in any way for their contributions; thank you indeed and God Bless You All.

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TABLE OF CONTENTS
Page Declaration..................................................................................................................... i Dedication ..................................................................................................................... ii Acknowledgements ...................................................................................................... iii Table of contents ......................................................................................................... iv LIST OF TABLES......................................................................................................... vi LIST OF FIGURES ..................................................................................................... vii List of abbreviations and symbols ............................................................................... viii Abstract........................................................................................................................ ix CHAPTER ONE ............................................................................................................ 1 Introduction ................................................................................................................... 1 1.1 background of the study ..................................................................................... 1 1.2 Statement of the study problem .......................................................................... 2 1.3 Objectives of the study ....................................................................................... 2 1.4 Hypotheses ........................................................................................................ 2 1.5 Justification of the study ..................................................................................... 2 CHAPTER TWO ........................................................................................................... 4 LITERATURE REVIEW ................................................................................................. 4 2.1 Reproductive methods used to enhance goat production ................................... 4 2.1.1 Selective natural breeding............................................................................... 4 2.1.2 Artificial insemination ..................................................................................... 5 2.1.2.1 Techniques of AI in goats................................................................................ 6 2.1.2.2 Timing of AI in goats ...................................................................................... 7 2.1.2.3 Factors that affect successful artificial insemination ....................................... 7 2.1.2.3.1 Sexual maturity in males ............................................................................. 8 2.1.2.3.2 Semen collection ......................................................................................... 9 2.1.2.3.3 Semen processing ..................................................................................... 10 2.1.2.3.4 Preservation of semen............................................................................... 11 2.2 Applied clinical physiology of the female goat reproduction .............................. 11 2.2.1 Signs of estrus (heat) and estrus detection in goats ...................................... 11 2.2.2 Estrus synchronization in goats..................................................................... 12 2.2.2.1 Principle of estrus synchronization in goats .................................................. 13 2.2.2.2 Advantages of estrus synchronization in goats ............................................. 13 2.2.2.3 Methods of estrus synchronization ................................................................ 13 2.3 Pregnancy diagnosis ........................................................................................ 15 2.4 Summary of the literature review ...................................................................... 16 CHAPTER THREE...................................................................................................... 18 Materials and methods ................................................................................................ 18 3.1 Description of the study area ............................................................................ 18 3.2 Study design and sampling procedure .............................................................. 18 3.3 Semen harvest ................................................................................................. 20 3.4 Semen assessment .......................................................................................... 21 3.5 Dilution of semen.............................................................................................. 22 3.6 Cervical AI of does ........................................................................................... 23 3.7 Data collection and analysis ............................................................................. 24

iv

CHAPTER FOUR........................................................................................................ 25 RESULTS ................................................................................................................... 25 CHAPTER FIVE .......................................................................................................... 28 DISCUSSION.............................................................................................................. 28 CHAPTER SIX ............................................................................................................ 32 CONCLUSIONS AND RECOMMENDATIONS............................................................ 32 6.1 Conclusion ....................................................................................................... 32 6.2 Recommendations ........................................................................................... 32 References ................................................................................................................. 34 APPENDICES ............................................................................................................. 39

LIST OF TABLES
Page Table 1: Pregnancy diagnosis techniques ..................................................................... 16 Table 2: The age of goats as shown by dentition .......................................................... 18 Table 3: Experimental design ........................................................................................ 20 Table 4: Estimating semen quality by observation ......................................................... 22 Table 5: Estimating semen density................................................................................ 22 Table 6: Response of does to estrus synchronization treatments .................................. 25 Table 7: Relationship between age, body condition scores (BCS) and conception rate ............................................................................................................ 27 Table 8: Effect of method of estrus synchronization and breed of buck on conception rate ............................................................................................................ 27

vi

LIST OF FIGURES
Page Figure 1: Effect of age group on conception rate ........................................................... 26 Figure 2: Effect of body condition score on conception rate .......................................... 26

vii

LIST OF ABBREVIATIONS AND SYMBOLS

%
/

Percentage Feet Artificial insemination Artificial vagina Body condition score Cubic centimeter controlled internal drug releasing device controlled internal drug releasing devices Centimeter Conception rates And the others Grams International units Milligram Mill litre Degrees Degrees Celicius P value Pregnancy diagnosis Poverty Eradication Action Plan Pairs of permanent incisor teeth Plan for Modernization of Agriculture Pregnant mare serum gonadotropine Ultra high temperature Uganda shillings Chi square

AI AV BCS Cc CIDR CIDRs Cm CR et al G IU Mg Ml

o o

P PD PEAP PI PMA PMSG UHT Ushs X2

viii

ABSTRACT
The conception rates of estrus-synchronized indigenous Ugandan goats with fresh semen cervical artificially inseminated were not known. The study aimed at establishing the effects of synchronization technique, type of semen, age and body condition of does on conception rate. A total of 160 Ugandan indigenous goats were randomly selected from Kabukye and neighboring villages. Does were randomly divided into four equal groups of which; two groups were treated for 17 days with 45 mg progesterone impregnated sponges and the other two groups with Controlled Internal Drug Releasing device (CIDR) containing 3 g progesterone. All does received intramuscular injections of 200 IU Pregnant Mare Serum Gonadotropine (PMSG) at sponge or CIDR removal. During the synchronization period, 2% sponges and 5% CIDR were lost before day 17. Semen was collected from one Boer and one Toggenburg bucks, and was evaluated to ensure that the bucks were fertile. Cervical artificial insemination (AI) was performed at a fixed time (48 and 56 hour) following progestagen withdrawal in (n = 37 sponge and n = 36 CIDR) treated does with fresh Boer semen, and in (n = 40 sponge and n = 36 CIDR) treated does with fresh Toggenburg semen. Pregnancy diagnosis done by observation of non return to estrus from day 17 to 22 post AI indicated that 93 (62.4%) does did not return to estrus. Ultrasound pregnancy diagnosis performed at day 48 post AI revealed conception rate 23 (62.2%) in does treated with sponge, 200IU PMSG, and inseminated using Boer fresh semen; 24 (66.7%) in does treated with CIDR, 200IU PMSG, and inseminated using Boer fresh semen; 27 (67.7%) in does treated with sponge, 200IU PMSG, and inseminated using Toggenburg fresh semen; and 18 (50.0%) in does treated with CIDR, 200IU PMSG, and inseminated using Toggenburg fresh semen. Levels of significances were tested using Chi-square test. There was no significant difference (P = 0.287) between use of sponge and PMSG, 50; (64.9%) or CIDR and PMSG 42; (58.3%) on conception rates. There was no significant difference (P = 0.430) between the bucks which provided semen on conception rate (Boer semen 47 (64.4%) and Toggenburg semen 45 (59.2%). The age determined by the number of pairs of permanent incisor teeth the does had, significantly (P < 0.05) influenced the conception rates across all the age groups. The nutritional status (body condition score measured on a scale of 1 - 5) significantly (P < 0.05) influenced the conception rate. These results indicated that, the use of sponge/PMSG and CIDR/PMSG intra vaginal progestagen treatments are equally efficient in synchronizing estrus in Ugandan indigenous goats. Chi-square test between semen used (P = 0.430), chi-square test between synchronization method (P = 0.287); indicated

ix

no significant difference between fertility of the Boer and Toggenburg. The overall fertility results (61.7%) are satisfactory; will increase/improve with experience.

CHAPTER ONE Introduction 1.1 background of the study

Artificial insemination (AI) for cattle is a routine and very popular exercise in Uganda. This technique is, however, still new for goats in Uganda even if it has been applied widely in several countries such as France, India, and Philippines (Leboeuf, 1992; Dao, 1996; Leboeuf et al., 1998; Pradip, 2004; Sohnrey and Holtz, 2005). Recently, interest in goat production has increased in the rural areas, where the climate is suitable for this kind of production. The indigenous goat breeds in Uganda are the Small East African, Mubende and Kigezi goats (Mason and Maule, 1960). These goats are being crossbred with the imported breeds for example the Boer goat, Toggenburg and Saanen. However, the numbers of bucks of those imported breeds are not adequate for the breeding program, importation of live goats is expensive, and importation of top quality bucks in large numbers is almost impossible. Good bucks are needed for the breeding, especially in the rural areas where the poor households desire to improve their livelihood (nutrition and incomes) through improving their goat productivity. According to Haenlein et al. (2008) artificial insemination (AI) has several advantages over natural service. It eliminates the necessity of keeping one or several bucks on the farm (depending on herd size). Buck related costs of feeding, housing, fencing and labor are eliminated when AI is used. AI can increase the rate of genetic improvement in herds, as long as superior bucks are consistently selected. In natural service, the prospective breeder has only the buck' pedigree to rely on, whereas AI bucks are progeny-tested for their s genetic transmission ability of milk and fat percentage, weight gain, conformation, etc. AI allows breeding of different portions of the herd to different bucks. Young does may be bred to not yet proven but high potential bucks, while the majority of the herd can be bred to proven high quality bucks. AI permits breeding of many does on one day when synchronization is practiced. The danger of transmission of diseases or parasites is also greatly reduced. The time of breeding can be more carefully regulated, and the owner knows exactly when the doe was bred, as opposed to pasture servicing by a buck that is allowed to run with the herd. AI induces good recordkeeping of dates of heat, breeding, pedigrees, etc; which aid in herd improvements as well as enabling the owner to make better management decisions.

1.2

Statement of the study problem

A number of exotic goats have been imported into Uganda in a bid to improve the

productivity of the indigenous breeds through crossbreeding programs. The importation of live animals is expensive, irregular, and may fail to capture top proven genetics. These factors lead to low levels of genetic improvement of the indigenous breeds, inbreeding and failure to implement the program over a wide area coverage due to lack of adequate number of bucks. With increased demand for quality products both for home use and trade, alternative methods for achieving higher levels of goat productivity need to be sought. Artificial insemination in goats in Uganda as a breeding method capable of contributing to the improvement in the productivity of indigenous goat breeds is at a low level of use. Although no documentations are available, it has been alleged goat inseminations achieved very poor conception rates and hence not worth to promote. Therefore, for adoption of goat AI as a method through which improving goat productivity can be achieved; the conception rate to cervical AI method was investigated to identify action of procedures that yield the highest conception rates.

1.3

Objectives of the study

The present study was carried out with the objective of comparing cervical AI conception

rates (CR) of indigenous Ugandan goats (the Small East African, Mubende and Kigezi goats) to a dairy breed and meat breed semen following estrus synchronization using intra-vaginal sponges and Controlled Interval Drug Release (CIDR) devices containing progesterone.

1.4

Hypotheses
The study hypothesized that: Estrus synchronization by use of sponges and CIDR yields similar conception rates. Conception rates to fresh Toggenburg semen and Boer semen do not differ. Conception rates are dependant on the age of the does inseminated. Conception rates are dependant on the body condition score of the does inseminated.

1.5

Justification of the study

The findings of this study will contribute to the planning, development and use of this

breeding biotechnology in spearheading the improvement of goat productivity in Uganda. The method of cervical artificial insemination utilizing fresh semen will enable development of skills for collection and use of fresh semen from the few proven elite bucks available to cover many more female goats than those each buck can serve in a given period of time. This will contribute to increased genetic improvement of the indigenous goats, and hence their productivity when favorable environment and management are achieved. Field

veterinary staff and eligible farmers can be trained and equipped to utilize this breeding method in the Plan for Modernization of Agriculture (PMA) and Poverty Eradication Action Plan (PEAP). Synchronization will enable programmed breeding, batch management of offspring and achieving uniform batches for sale in market-led production systems. Furthermore, the inadequate availability of liquid nitrogen, high costs of cryopreservation equipment and poor infrastructure favors the use of fresh semen in improving goat productivity in the remote rural areas of Uganda.

CHAPTER TWO LITERATURE REVIEW 2.1 Reproductive methods used to enhance goat production
The goat is a source of meat, milk, skin and fibre. Cross breeding for milk and meat, is a common practice. If we improve environment, management, nutrition, selection of breeding and disease control, we can improve the economics of goat farming. Improvement in reproductive efficiency will increase the economic viability of goat enterprise. The reproductive efficiency depends on the embryo/fetal development, litter size, survival viability, growth of newborn, suckling period, age at puberty and duration of reproductive life (Panhwar, 2007). Reproductive efficiency can be improved by the use of modern reproductive tools which result into more kids per genetically superior bucks and does.

2.1.1 Selective natural breeding

Selective natural breeding is whereby of animals having desirable characters are mated. In selective natural breeding, a doe (female goat) in heat is bred to a buck (a male goat) (Irvine, 1995). Advantages of natural breeding include simplicity in that goats have been breeding naturally for centuries. The doe owner selects which buck to use, and the buck takes care of the rest. The method offers cheap breeding cost because if a farmer does not own a buck, other goat owners could allow breeding to their buck for a fee (Irvine, 1995). At Kabukye in Kamuli district of Uganda, this fee ranges from around Uganda shillings (UShs.) 0 to 500 (US$ 0.25) depending on the quality of the buck being used. The buck owner uses his buck as a source of income by breeding other farmer' does. A doe is in heat s from 12 to 36 hours, and after breeding naturally semen can live for about 24 hours (Pradip, 2004). It' not essential to breed at exactly the correct timing, and it' easy to repeat the s s breeding for a few days (Panhwar, 2007). The disadvantages of natural breeding include difficulty in managing a buck (Irvine, 1995). Bucks require sturdier fences than does, and are usually housed away from does. During breeding seasons, bucks have some unusual behaviors such as urinating on their beards. This creates an offensive smell that attracts does which seem to appreciate it. There is lack of variety of bucks used. Irvine (1995) further explained that use of one buck or a few bucks present on the farm limits the selection of breeding bucks to be used for breeding. Selection of breeding bucks is limited by distance to be traveled to access bucks if the bucks belong to some other farms. Bucks frequently cost more than does and not readily available, this limits the number of bucks kept on the farm. Consequently, this may lead to inbreeding. Bucks also eat more than does, and destroy their fences more frequently.

2.1.2

Artificial insemination
In artificial insemination (AI), diluted fresh or frozen semen from a buck is

mechanically inserted into the cervix or uterus of a doe in heat (Irvine, 1995). Advantages of artificial insemination include elimination of the necessity of keeping one or several bucks on the farm (Haenlein et al., 2008). Costs of feeding, housing, separate fencing and labor are also eliminated. Once you have the necessary equipment, fresh or frozen semen is much less expensive than the costs of housing and feeding a buck (Irvine, 1995). AI can increase the rate of genetic improvement in a herd, as long as superior bucks are consistently selected for semen collection. It is therefore possible to quickly improve the quality of your herd using such resources which would have been spent on the management of the buck. In natural service, the prospective breeder has only the buck' pedigree to rely on (Haenlein et s al., 2008), whereas AI enables rapid evaluation of the growth rate and carcass characteristics of a bucks progeny at slaughter. Pedigree bucks can be progeny-tested (McDougall, 1995) for their genetic transmission ability of milk and fat percentage, weight gain, type conformation, etc, (Haenlein et al., 2008). AI also enables the widespread use of out-standing bucks and dissemination of valuable genetic material even to small farms (Hafez, 1993). Furthermore; AI facilitates progeny testing under a range of environmental and managerial conditions, thereby further improving the rate and efficiency of genetic selection (Hafez, 1993). AI is a method of importing new genes, even breeds, into a country, while reducing the risk of importing diseases (Peacock, 1996). AI has the potential to reduce disease transmission between breeding animal. This includes not only the venereal and reproductive diseases which are transmissible through mating, but other pathogens spread via contact, (McDonald and Pineda., 1989). AI allows breeding of different portions of the flock to different bucks. Young does may be bred to not yet proven but high potential bucks, while the majority of the flock can be bred to proven high quality bucks. AI also permits breeding of many does on one day when synchronization is practiced, which enables the time of breeding to be more carefully regulated, and the owner knows exactly when the doe was bred, hence good record keeping (Haenlein et al., 2008). In addition, AI enables compacted kidding that saves labor and simplifies husbandry and management routines (McDougall, 1995), as opposed to pasture servicing by a buck allowed to run with the herd. Record keeping also aids in herd improvements and enable the owner to make better culling decisions (Haenlein et al., 2008). AI allows one quality/proven buck to serve more does during a period of time than that buck could with natural service (McDougall, 1995).

2.1.2.1

Techniques of AI in goats

There are 3 main techniques of AI in goats, namely: blind AI, cervical AI and laparoscopic intrauterine AI.

Blind AI
Blind insemination consists of deposition of semen into the does anterior vagina with a pipette without an attempt to locate the cervix with the help of a light source (McDougall, 1995). This method requires very little equipment and skills. A dose of 500 million sperm from fresh semen per doe is used to give conception rates of 50 60 % (McDougall, 1995). Usually low conception rates are achieved with this technique even after using neat semen. This technique is practically useless if frozen semen is to be used (Pradip, 2004).

Cervical AI
According to Panhwar (2007), cervical insemination is carried out by lifting the hind legs of females at an angle of 60 from the ground. In this method it is necessary to locate the cervix with the help of a speculum and a source of light or the doe should be placed so as to allow the use of sunlight for the location of the cervix and insemination (Morrow, 1986). If the doe struggles, the helper should hold its body between her/his legs and hold its hind legs firmly, while the insemination is carried out. The inseminator should gently introduce a lubricated speculum into the vagina to determine the stage of estrus. If a large quantity of vaginal mucus is present, preventing the correct location of cervix, the assistant should tilt the animal down to allow the mucus to run out with the aid of the speculum. If the stage of oestrus is correct, the inseminator should hold the speculum while another person loads the pipette with semen and hands it carefully to the inseminator. The inseminator should then attempt to pass the tip of the pipette through the cervix carefully with a rotary motion. If this is achieved, then the pipette should be withdrawn a little and the semen deposited (Pradip, 2004). In 30 - 40% of does, it is possible to pass the pipette through the cervical folds and deposit the semen into the uterus. In the rest, semen is deposited into the cervix after penetration of 2 3 cervical folds (McDonald and Pineda., 1989). The doe should then be gently returned to the flock. Take care that no buck has access to the inseminated doe till estrus is over. Otherwise the buck may mate with it. Ensure that no urine or faecal matter contaminates the tip of the pipette during insertion as this can reduce conception rates. For every insemination a fresh, sterilized test tube should be used for thawing of semen. The speculum should be wiped clean with cotton wool and the pipette rinsed out with distilled water and then shaken dry between inseminations (Panhwar, 2007).

According to McDougall (1995), cervical insemination requires slightly more equipment than blind AI, and a conception rate of 60-70% can be achieved with 250 million sperms of fresh semen per doe. This technique of AI is suitable for farm and village use. It can be carried out by the goat owner as it is quite simple and effective if done with care. The doe in estrus should be removed from the flock. It should be inseminated about 12 hours after onset of estrus, i.e. does detected to be in estrus in the morning should be inseminated in the evening and those detected in the evening should be inseminated the next morning (Pradip, 2004).

Laparoscopic intrauterine AI
This involves bypass of the cervix by using a laparoscope to visualize the uterus in the abdomen, then injecting semen directly into the uterus with a needle pointed pipette (McDougall, 1995). This is a high-tech procedure performed using expensive equipment, requires certain amount of skills and therefore performed by veterinarians only. This technique enables use of frozen semen at a dose of 20 million live sperm per doe with conception rates of 60-70% and a conception rate of 80-90% with fresh semen (McDougall, 1995).

2.1.2.2

Timing of AI in goats

About 30 to 36 hours after the onset of estrus, the ovum is released from the ovary as reported by Pradip (2004). The ovum remains viable in the female reproductive tract for 12 to 24 hours after release. Following insemination, the sperm remains viable in the female reproductive tract for at least 24 hours. Pradip (2004) further reported that cervical AI should be carried out 12 to 18 hours after onset of estrus agreeing with Morrow (1986). This means that semen should be present in the reproductive tract when the ovum is released. Either fresh or frozen semen may be used. If AI is delayed, conception rates will be poor due to the aging of the ovum. When the doe comes into heat the vaginal mucous membrane begins to get pink due to an increase in blood supply. At the same time there is a vaginal discharge. Later the vagina starts to look pink and shiny. The best results are obtained if AI is carried out at this stage. If the doe is mated naturally then the vagina begins to look cloudy. When estrus is over, the pink colour becomes paler and the vagina looks whitish with tiny wrinkle. 2.1.2.3

Factors that affect successful artificial insemination

Artificial insemination can be a successful technique in goat production. However, the

success would depend upon the understanding of applied reproductive physiology such as selection of breeding goats, semen collection, semen processing, optimum number of live

and normal spermatozoa per inseminated goats, estrus behavior, stage of heat and insemination technique, (Panhwar, 2007). A reliable means of inseminating goats would be most advantageous, not only for commercial breeders interested in genetic improvement of their stock, but also for smaller producers who cannot afford a breeding buck of their own, and for persons who have a problem with the smell of a billy goat in their yard. In France where systematic genetic improvement of dairy goats is pursued, AI is part of the management routine (Sohnrey and Holtz, 2005). When appropriately conducted, AI using fresh semen produces conception rates comparable to those obtained with natural breeding (Leboeuf, 1992; Leboeuf et al., 1998) With frozen-thawed semen, lower conception rates generally are encountered (Sohnrey and Holtz, 2005). For the French situation, conception rates of 60 to 65% have been reported (Leboeuf, 1992; Leboeuf et al., 1998). In other circumstances including does under stress (Wayne 2002), use of fresh poorly cooled or frozen semen, inappropriate timing of insemination and improper deposition of semen poor conception rates are frequently experienced (Teresa, 2000). Unsatisfactory pregnancy rates are a common complaint when inseminating goats using cryopreserved semen through the traditional cervical method (Evans and Maxwell, 1987). Superior and more consistent pregnancy/conception rates may be accomplished when does are inseminated laparoscopically (Morrow; 1986, Moore et al., 1988 and Ritar et al., 1990). The advantage of laparoscopic insemination is that semen is deposited closer to the site of fertilization. However, this is a high level technology and high cost procedure which can be successfully done on station or established farms, but not on peasant farms settings.

2.1.2.3.1

Sexual maturity in males

According to Panhwar (2007), sexual maturity is the age or stage when an organism can reproduce; brought about by maturing of the reproductive organs and the production of gametes. Sexual activity in male is determined by placing them with estrus female daily during morning and evening hours from the age of 6 months and onwards. Males showing active courtship tendency for sexual mount or natural service are separated from the rest of flock (Morrow, 1986). The remaining males are given further chance for expression of sexual desire, till the majority of them have shown complete sexual mounting. Immature bucks of all breeds produce poor quality semen. As the bucks mature, they begin to produce high quality semen with regard to sperm motility and morphology. The sperm concentration per milliliter

and total number of sperm per ejaculate increase gradually up to two years of age (Youngquist, 1997).

2.1.2.3.2

Semen collection

Bucks are handled basically the same way as bulls for semen collection. Three basic methods may be employed, but all three require an artificial vagina. An artificial vagina is a double walled device with an opening at one end and collection tube at the other. The inner lining holding warm water should be coated with a light application of water soluble lubricating jelly (McDonald and Pineda, 1989). Good quality semen sample from breeding goats may be collected once daily from native and half breeds, and once or twice in quick succession (3-5 minutes) after every alternate day from exotic and higher cross-bred goats (Panhwar, 2007). In the first method, sexually active bucks should be trained for semen collection using artificial vagina. Daily during morning hours they are exposed to an estrous doe fixed in a service crate (Panhwar, 2007). The buck is allowed to mount a doe, with the semen collector manually diverting the buck' penis into the artificial vagina. The semen s collector does not touch the penis directly; instead the collector directs the penis into the artificial vagina by grasping the buck' prepuce (Morrow, 1986; McDonald and Pineda, s 1989). After ejaculation (usually 0.5 to 1.0 cc) has occurred, the artificial vagina is removed and tipped so that semen runs into the collection tube. This method may require practice and adjustment by both the buck and the collector before good samples are collected (Haenlein et al., 2008). The second method requires training the buck to mount a dummy instead of a live doe. The method follows the same procedures as the first method. Mounting may be facilitated by applying vaginal mucus scrapings of a doe that is in heat to the dummy, at least during the training process (Haenlein et al., 2008). The third method involves the use of an electro-ejaculator. The bucks rectum is emptied by a warm-water enema (Morrow, 1986). The buck is not required to mount an object, although an artificial vagina should still be used for semen collection. An electrode unit, which has a number of contact rings, is inserted into the buck' rectum. An electric s stimulation causes ejaculation. This technique generally results in good samples in quantity and quality. However, the sperm concentration of the sample is generally low. This method does not require extensive training, and will allow collections from bucks that may refuse or are unable to mount and serve an artificial vagina (Haenlein et al., 2008).

2.1.2.3.3 a)

Semen processing

Semen processing is done in 2 steps: semen evaluation and extension.

Semen assessment
Irrespective of semen collection procedure and frequency of semen collection, each

ejaculate is examined for its quantity and quality. Parameters considered in semen evaluation includes volume, color, consistency, motility, sperm density, and total sperm number per ejaculate, per cent live and normal spermatozoa. These parameters are generally correlated positively with fertility (Morrow, 1986; Panhwar 2007). According to Panhwar (2007), good quality semen is ivory to cream colored with thick consistency containing excellent wave motion. Sample containing less than +4 motility score with a density less than 3000 million sperm and less than 80 per cent live spermatozoa should not be used for breeding purposes.

b)

Extension of semen
Immediately after collection and physical evaluation, semen samples should be

extended to the correct density using available diluent, for example UHT skimmed milk, egg yolk citrate glucose and tris, (McDougall, 1995; Pradip 2004). During semen dilution, both the semen and the diluents should be at 30 Always add the diluent to semen. When C. extending semen, it is essential that the minimum number of spermatozoa is present in each dose in order to achieve an acceptable conception rate. The minimum number will vary depending on a number of factors, for instance 100, 250 and 500 million sperm for does in natural estrus, sponged in the breeding season and sponged out of the breeding season, respectively (Morrow, 1986; McDonald and Pineda, 1989; McDougall, 1995; Pradip 2004). For goats that have been synchronized the use of higher doses of PMSG requires that more live sperm be included in each doe dose. Where greater than 500 IU of PMSG is used, the 500 million sperm should be inseminated into each doe. If excess semen is available, then the ejaculate is split amongst the does available. The volume of semen inseminated is also critical in order to ensure that the semen remains in the cervix of the doe. The volume inseminated must not be more than 0.25 ml per doe. The ideal insemination volume is 0.1 ml, i.e. 0.1 ml should contain the required amount of spermatozoa. If only a small amount, e.g. 0.2 ml, of diluent have been added to the semen sample, then semen is best used immediately. This requires that when using a dose of 0.1 ml per doe, inseminate does in 1 hour after collecting the semen (McDougall, 1995). According to McDougall (1995), if semen is to be diluted and held for 46 hours then the semen should be diluted down to 1000 million sperm per ml with UHT skim milk. Trials by McDougall (1995) have revealed that

10

acceptable conception rates are achieved at this density when semen is stored for 4 -6 hours.

2.1.2.3.4

Preservation of semen

Goat semen can be stored successfully up-to 48 hours at 4-7 with satisfactory C sperm motility and fertility (Panhwar, 2007). Semen preservation by this procedure when done carefully, the entire cooling process is completed in 1.5 to 2 hours. Long term storage of goat semen is possible through deep freezing technique (Morrow, 1986). According to McDonald and Pineda, (1989) and Panhwar (2007), milk, tris and egg yolk citrate glucose diluents containing glycerol as the cryoprotective agent are used for freezing buck semen either by straw or pellet method. Straws filled with diluted semen and frozen in liquid nitrogen vapor can be stored for years in liquid nitrogen (-196 Post-thawing motility and fertility of C). frozen goats semen is relatively lower than the freshly diluted semen (Panhwar, 2007). Alternatively, semen may also be frozen into pellets by dropping cooled semen into depressions drilled or scratched into solid carbon dioxide (dry ice). This is a convenient method for freezing, but semen treated this way is more difficult to thaw and use (Morrow, 1986; Pugh, 2002).

2.2

Applied clinical physiology of the female goat reproduction

2.2.1 Signs of estrus (heat) and estrus detection in goats

For AI to be successful, it is necessary to detect the signs and the stages of estrus. According to McDonald and Pineda (1989); Hafez (1993) and Pradip (2004), the signs of estrus in goats include: rise in body temperature, bleating and frequent wagging of the tail, marked reduction in milk yield in milking does, mounting others and standing still when mounted by other does. There is also clear or white discharge from the vagina which may be seen stuck to the tail and around surrounding region, swollen and reddish vulva, and if there is a buck around, a doe in estrus will try to approach it or keep looking at it. Usually a person working with the does will be able to detect a doe in estrus. However, if there is a large flock of goats, it may not be possible to observe individual goats in estrus. In this case it is necessary to use heat detection aids such as use of bucks or fellow does (teasers) to detect females on heat. Examples of use of heat detection aids are outlined below as described by Pradip (2004): I. An intact buck is brought into the flock. The buck will follow the doe in estrus. However, there is a chance that the buck will mate with the doe if it is not complexly controlled.

11

II.

Using a buck with an apron tied on the abdomen of the buck to cover the penis. The apron is made of cloth measuring 60 cm by 45 cm with strings on the four sides to tie it properly to prevent mating. The apron should be washed daily and checked for holes or tears to avoid unwanted mating.

III.

Using a vasectomized buck that is six to nine months old. After a vasectomy, it is necessary to wait for three weeks before using the buck for estrus detection. Two or three vasectomized bucks are needed for estrus detection in a flock of fifty does.

IV.

Using a buck after tying a string around its prepuce, a technique used by shepherds. The buck can urinate but cannot protrude the penis from the prepuce, thus preventing mating. The other end of the string is tied around the testicles to hold it in place.

V.

Using an androgenized doe causing male-like sexual behavior in the doe for the purpose of detection of estrus. Injections of the male hormone testosterone (50 mg) are given to a doe every alternate day for twenty days. The treated doe will learn to detect estrus if it observes a buck doing so. Once estrus is detected in a flock, the marked/identified doe should be removed from the flock so that the teaser will detect any other doe that may be in estrus. If this is not done, it may pay attention only to the first doe and ignore the others that may be in estrus (Pradip, 2004). Animals tend to be more sexually active around dawn and dusk, which also corresponds to the coolest times of the day as well as times when human activity tends to be minimal. For whatever reason, observations of at least 30 minutes each at dawn and dusk will permit detection of a large percentage of does in heat. The best times to watch are first thing in the morning before feeding and/or milking and late in the evening after milking and/or feeding have occurred. The large flocks at pasture are best gathered in a small area (a corner of the field will work) to observe and to increase the interaction between does (Nix, 2002).

2.2.2 Estrus synchronization in goats

Synchronization of estrus is a technique, which is used to bring large number of animals in a flock into overt heat at the predetermined time (Panhwar, 2007). Manipulation of the estrus cycle in goats facilitates scheduling the breeding of does to be naturally serviced or artificially inseminated (Ott et al., 1980 a; Armstrong, 1982; Bretzlaff and Madrid, 1985; William and Braun, 1985; Mgongo, 1988; Bretzlaff et al., 1991; Wildevs 1993). Estrus synchronization together with AI is extensively applied in the reproductive management of goats (Leboeuf et al., 1998).

12

2.2.2.1

Principle of estrus synchronization in goats

According to (Pradip, 2004), in a flock, any normal healthy doe which is not pregnant is either in the luteal phase or oestrogenic phase. Progesterone hormone concentration increases in the luteal phase. This hormone is produced by the corpus luteum. At every estrus, one or more ova are released from the ovaries and at each ovum release spot; a corpus luteum is formed in the ovary. In any flock, 70 80% of the goats will be in the luteal phase because the period of the luteal phase is longer than the period of the oestrogenic phase. The luteal phase can be shortened or lengthened using hormones and this technique can be used to bring a group of goats in estrous at the same predetermined time.

2.2.2.2

Advantages of estrus synchronization in goats

The technique offers an opportunity to increase the efficiency of animal production in different ways and this is useful in the implementation of an AI program (Pradip, 2004). It reduces the time needed for detection of estrus, and it helps in conjunction with a procedure for controlling the time of ovulation to permit insemination on a predetermined schedule (Panhwar, 2007). Timed breeding following estrus synchronization allows for parturition at suitable times to take advantage of niche markets, feed supplies, labor, and rising price trends (Morrow, 1986, Whitley and Jackson, 2004; Haibel, 1990; Shelton and Lawson., 1982). Kids born to does following estrous synchronization are roughly of the same age at any given time, which makes it easy to manage their feeding according to nutrient needs and compare efficiency of growth. Synchronization of estrous enables accelerated breeding and the production of a flock every eight months thus reducing the kidding interval (Hafez, 1993). Estrus synchronization is necessary to carry out an effective Embryo Transfer program (Morrow, 1986; Pradip, 2004). It reduces the mortality at the time of parturition by avoiding breeding during extreme seasons. Successful control of breeding helps in reducing the neonatal mortality, and permits weaning, fattening and marketing of uniform group of animals. This promotes improved management of animals of the same age which provide better market price (Panhwar, 2007).

2.2.2.3

Methods of estrus synchronization

Estrus synchronization can be achieved by using buck introduction effect (Morrow, 1986). Usually if there is no buck in the flock and a buck is introduced, a large number of does will come into estrus. This is because the buck releases hormones (pheromones) from glands located at the base of its horns (Pradip, 2004). To get this effect, there should be no buck in the flock or within sight, smell or hearing for at least three weeks. Once a vasectomized buck or a buck with an apron is introduced, the does come into estrus within 5

13

 6 days and may be inseminated based on estrus detection (Pradip, 2004). This is a good strategy to naturally synchronize breeding does at the start of the breeding season or it can be used to induce does into heat out-of-season. Pharmacological methods of estrous synchronization include the use of

progestogens and luteolytic prostaglandins. The luteolytic prostaglandin (PGF2 ) causes luteal resgresssion and eventually the doe comes to estrus. This approach is only applicable in cyclic females and hence prostaglandin-based systems are restricted for use during the breeding season in goats (Morrow, 1986). Because not all stages of the estrous cycle are similarly receptive to treatment, a double injection system 11 days apart is the most widely used approach in goats. The mean time from injection to behavioral estrus is 46 to 48 hours (Morrow, 1986 and Wildeus, 2000). Prostaglandin method is however, not so successful in goats and hence is not commonly used (Pradip, 2004). Progesterone or a progestagen analogue is used to synchronize estrus in does during the breeding and non-breeding seasons. Progestagens are used in form of progesterone impregnated sponge or CIDR (Controlled Internal Drug Release). The most common route of progestagen application in goats is intra-vaginal (Bretzlaff, 1997). The sponge or CIDR is introduced into the vagina of the doe using an applicator. The device has a nylon string attached to it that hangs outside the vulva. The device is usually inserted over periods of 9 to 19 days and is removed by pulling the string (Wildeus, 2000). After removal the device should be burnt or buried deep to prevent it being eaten by animals or misused in other ways (Pradip, 2004). At time of removal, PMSG is injected to each doe to enhance ovulation. The synchronized does will exhibit estrus simultaneously around thirty hours after the removal of the sponge or CIDR (Pradip, 2004). Other procedures for synchronization of estrus are administration of fluorogestone acetate (FGA) sponges and medroxyprogesterone / methylacetoxyprogesterone (MAP) treatment for 12 - 21 days (Romano, 2002; Romano and Benech, 1996; Romano and Fernande, 1997; Leboeuf et al., 1998; Romano et al., 2000). These require an intramuscular injection of PMSG at progestagen treatment withdrawal (Greyling and Van der Nest, 2000; Motlomelo et al., 2002). The does may get 11 days of treatment with FGA impregnated intravaginal sponges and an intramuscular injection of PMSG and a synthetic PGF2 analogue 48 h before or at sponge withdrawal (Baril et al., 1993; Freitas et al., 1996 a, b; Freitas et al., 1997; Leboeuf et al., 2003). Estrus synchronization can also be achieved by the oral administration of progesterone in form of melengesterol acetate (MGA) in feeds. The treatment period vary

14

between 5 - 18 days before introduction of the males, and may also include an injection of gonadotropin at or before withdrawal of progestogens treatment followed by introduction of the male (Evans and Maxwell, 1987; Haibel 1990). Another estrus synchronization approach is by use of artificial lighting to synchronize out-of-season breeding in goats by mimicking natural photoperiod-induced estrus (Walkden-Brown and Martin 1997). Controlling lighting requires confinement of females. Alternatively, treatment of bucks and does with melatonin (the hormone of darkness) which mimics the short-day photoperiod after exposure to natural long-day photoperiod can be used to alleviate the need for controlled lighting and associated changes in confinement area (Singh-Knights and Knights, 2005). It must be remembered that, the type of treatment used and likely rate of success will depend greatly on the extent of breed susceptibility to photoperiod and time of the year in seasonal breeders, physiological state, body condition score (BCS) and nutritional status. Also, conception and pregnancy rates for these procedures tend to be lower than breeding at natural estrus (Singh-Knights and Knights, 2005).

2.3

Pregnancy diagnosis
A number of tests; as shown in Table 1 are used for detecting pregnancy in goats.

The choice of the technique depends on the stage of gestation, cost, accuracy and speed of diagnosis (Pradip, 2004).

15

Table 1: Pregnancy diagnosis techniques

No. a) Technique Used Non-return to estrus. Description Post breeding/insemination non-return to estrus gives an idea of pregnancy status. During the breeding season, most of the does return to estrus within 17-23 days after fertilization failure. At the end of breeding season, nonreturns to estrus are considered to indicate pregnancy. b) c) d) Visual assessment Laparotomy. Visual assessment through abdominal ballottement shows advance stage of pregnancy. Needs surgery. It gives 90 - 95% accuracy in goats of 5 weeks gestation. Laparoscopy/Endoscopy Presence or absence of pregnancy can be detected by direct observation of the uterus and ovaries through laparoscopy/endoscopy. Pregnancy can be detected at 40 days of gestation in goats. e) f) g) Radiography. Ultrasonic technique. Vaginal cytology. It has limitation because it can only be performed in a laboratory/hospital. It is good and produces immediate results and can be adopted for field use. It is impracticable under field conditions, although pregnancy can be detected up to 95% of animals at 40 days of gestation. h) Hormonal assay. This test is based on the level of pregnancy-dependent hormones in blood, milk or urine. Radio immunoassay and competitive protein binding an ELISA technique are used for the detection of hormone. With this technique pregnancy is diagnosed at earlier stage showing accuracy of 95%.

Source: (Panhwar, 2007) 2.4 Summary of the literature review

Despite the method of reproduction used to enhance goat production, the health of the doe and its reproductive system are important factors that contribute to the productivity of the flock. When AI has been selected as the breeding method, the AI technique to be utilized must be logically selected. The technique of AI as well as the skill and experience of the person carrying out the AI are very relevant to conception rates achieved. The deeper in the cervix the semen is deposited during cervical AI the better the results. The quality of the semen and

16

number of viable spermatozoa present in the semen should be carefully adhered to as recommended for the technique to be used. Fresh or frozen can be used however, usually lower conception rates are achieved with frozen semen mainly because spermatozoa partially loose their vigour on freezing (Pradip, 2004). Timing of the insemination in relation to the onset of estrus is of utmost importance and hence accurate estrus detection is necessary. Survival of spermatozoa deposited into the reproductive tract of the doe and the simultaneous presence of a receptive ovum in the uterus are two vital factors on which conception depends. Therefore the success of AI also depends mainly on the time of insemination. Estrus synchronization enables carrying out AI at predetermined time and kidding at the same time. It should be noted that conception rates are usually lower in maiden does, increases up to a certain age and decline thereafter. Stress should be avoided at the time of and after AI. After AI the doe should be returned to the surroundings and flock to which it is accustomed. It should also receive the same feed in adequate quantities. There is a risk of early embryonic loss if the doe' blood sugar declines due to a deficiency in nutrition. Where doe breeding seasons s occur, conception rates are usually better when AI is carried out during the normal breeding season (Pradip, 2004). It is therefore important to carefully consider these and all the other factors mentioned in this study for successful implementation of cervical goat AI in Uganda.

17

CHAPTER THREE Materials and methods 3.1 Description of the study area
The present study was done in Kabukye and the neighbouring villages, Kitayunjwa sub-county in Kamuli district of Uganda. Kabukye is located at approximately N 00.897400 and E 033.122690, and at approximate altitude of 1,101 metre above sea level. Kamuli district is located in south-eastern Uganda, it extends from 00 - 56 North / 330 - 05 East up to 010 - 20 North / 330 - 15 East (Appendix I), at approximate altitude of between 941 and 1,101 metres above sea level. The district experiences a bimodal type of rainfall which is about 110 mm during the main season that extends from March to May and least during the months of August through October. The district is mainly covered with savannah vegetation with scattered remains of the equatorial forest cover which has been depleted over time. The district has annual mean temperatures range from 220 C minimum to 290 C maximum.

3.2

Study design and sampling procedure

A total of 160 female goats used in the study were the indigenous breed selected

randomly from the population of 275 goats in Kabukye and neighbouring villages. They had kidded at least once, thus were said to be breeder does. All the does were managed on a browsing/grazing system used by peasant farmers. All the goats were kept under the same management conditions during the study period. They were all subjected to ultrasound pregnancy diagnosis to identify the non-pregnant ones. Those found to be not pregnant were objectively conditionally scored using a condition score of five points; Score 1 was given to the very thin doe and score 5 was given to the very fat doe (Ebert et al., 2006). The does selected were those of BCS of 3 - 4. Each flock was assigned a research number, and each doe in the flock given a number. The selected does age was estimated using their dentition as shown in Table 2.

Table 2: The age of goats as shown by dentition

Age (in approximate years) 1 2 3 4 Source: (Haenlein et al., 1992) Type of teeth One pair of permanent incisor Two pair of permanent incisor Three pair of permanent incisor Four pair of permanent incisor

18

Both purposive and stratified sampling methods were used in the selection of the does. The does were grouped and stratified for age and body condition. The sample size of the flock used in the study was determined using the formula by Morgan and Krejcie (1970): S= Where: S= N= P= D= X =
2

X2 N P (1 P) D2 (N 1) + X2 P (1 P)

required sample size. population size of 275 does. population proportion assumed to be 0.50 as it yields maximum sample size degree of accuracy at 0.05. table value of Chi square given as 3.841 at 95% confidence level.

required.

From the formula above, the sample size was calculated thus: S= 3.841 x 275 x 0.5 (0.5) (0.05)2 (274) + (3.841 x 0.05 x 0.5) = 264.06875 0.685 + 0. 96025 = 160.77

But S is taken to be 160. As shown in Table 3 below, the 160 selected does were randomly divided into two hormonal treatment groups of 80 does each. The two groups were estrous-synchronized using either 45 mg progesterone impregnated sponges (Syncro-part, Ceva, France) or CIDR containing 3g progesterone (EAZI-BREEB CIDR Sheep and Goat Device, Pharmacia and Upjohn, Australia) intra-vaginal inserts. On the 17th day, an injection 200 IU of pregnant mare serum gonadotropine (PMSG) hormone (PMSG-Intervet, Intervet, UK) was given at withdrawal of implant to enhance ovulation. Estrous detection was done using a vasectomised buck and bucks with aprons. For each hormonal treatment, the does were randomly divided into two groups for each hormonal treatment then given double intra-cervical AI using fresh semen of either Boer or Toggenburg bucks. The interval from

19

treatment withdrawal to first AI was approximately 48 hours, while the interval between the first and second AI was approximately 8 hours. Pregnancy diagnosis was done first by observation of non-return to cycling from day 17 to 21 post-insemination, then at 48 days following insemination by use of ultrasound (Dynamic Imaging, Dynamic Imaging, Scotland, UK).

Table 3: Experimental design

Treatment 45 mg progesterone impregnated Sponge + 200 IU PMSG Boer semen Toggenburg semen Total 40 40 80 CIDR containing 3 g progesterone + 200 IU PMSG 40 40 80 80 80 160 Total

3.3

Semen harvest
Semen used for intra-cervical insemination was collected using an artificial vagina on

the same day of insemination as described by (McDougall, 1995). Briefly, the artificial vagina (AV) was prepared by placing the dry liner in the centre of the outer casing with the ends folded back over the casing and tightly fixed with 2 thick rubber bands so that there were no water or air leaks. Using a hand spray bottle, the inside of the liner was sprayed with surgical methylated spirit (Mavids surgical spirit, Mavid Ltd, Uganda) to sterilize it. The AV was half filled with warm water (60 70oC) immediately before collection by slowly introducing the water through the tap on the AV using a 50 ml syringe. The hot water evaporated the alcohol from the liner. A semen collection glass that had been pre-warmed to 37oC was inserted into one end of the AV to make a tight fit. When all the alcohol had evaporated, air was blown into the assembled AV until the lumen was tight. The lumen of the AV was then lubricated with KY jelly (KY jelly, Johnson and Johnson, UK) to reduce friction. Following this, a padded cover was put over the sperm glass to keep it warm and the AV was held horizontally to keep the entrance warm as well, while keeping the tap on the AV pointed down towards the ground. As the buck held by an assistant was teased and the buck jumped on the doe, its preputial sheath was intercepted using the free hand that directed the penis into the AV as

20

the buck thrusted and ejaculated into it. As the buck dismounted, the AV was withdrawn, held upright and the tap was released to expel the air and allow semen to flow into the sperm glass, while holding onto the sperm glass with foil. The semen collected was placed into the water bath which was pre-set at 37oC, while the top of the sperm glass was covered with a foil. The average time Boer and Toggenburg semen was kept from time of collection to the beginning of insemination was 12.8 and 23.6 minutes respectively. The difference in time was because the Boer semen was always collected after that of the Toggenburg had been collected.

3.4

Semen assessment
Semen was evaluated by comparing with the following standard parameters

described by McDougall (1995): Volume Color Density Motility % abnormal 0.5 1.5ml ivory yellow 2,500 5,000 million sperm/ml 80 95% 5 -25%

Using a pipette, a drop of semen was transferred onto a warm microscope slide and viewed under low power (X100 magnifications) for swirl movements. Swirl movement were graded from zero (totally no movements) to five (very high movements easily observed even with unaided eyes) (McDougall, 1995). The higher the wave movements, the more concentrated the semen (Pradip, 2004). Semen was accepted for cervical AI when the swirl was 4 5 and sperm seen moving in the sperm glass. If swirl movement was poor, then the semen was subjected to a further examination for abnormal or dead sperm with the aid of a microscope. Before the evaluation for live spermatozoa, the sample was evaluated for mass activity and individual progressive motility of spermatozoa. During mass activity evaluation a drop of freshly collected semen was placed on a clean microscope glass slide and examined under the microscope to observe mass activity of the spermatozoa in the form of wave motion. The higher and more concentrated the wave movements, the better the quality of the semen was. Individual progressive motility of spermatozoa was done to observe individual spermatozoa movement. The proportion of spermatozoa showing motility by swimming straight in one direction or showing linear movement was observed. Further, the concentration of the semen was determined by observation of its density and colour. Following evaluation, the semen was graded as indicated in Tables 4 - 5 below as described by McDougall (1995).

21

Table 4: Estimating semen quality by observation

Grade 1 2 3 4 5 Source: (McDougall, 1995)

Swirl No movement Can see sperm moving but no waves Slow wave motion Moderate wave motion Very rapid wave motion

% Motility 0 20 30 45 50 70 75 80 85 95

Table 5: Estimating semen density

Grade 0 1 2 3 4 5 Source: (McDougall, 1995)

Density Watery Skim milk like Milky Thin creamy Cream Thick creamy

No. per ml (in millions) 0 250 500 1000 1500 2000 2500 3500 4000 5000 5000 6000

3.5

Dilution of semen
Semen was diluted in tris buffer extender (containing 80 ml tris and 20 ml egg yolk) to

achieve the essential minimum number of spermatozoa in each dose in order for an acceptable conception rate (Pradip, 2004). The amount of diluent that was added to the semen was calculated as described by McDougall (1995). Briefly, the volume, motility and density of the ejaculate were first recorded. The number of motile spermatozoa in the ejaculate was calculated by multiplying the volume x density x motility, and the number of doe doses by dividing the number of spermatozoa in the ejaculate by the number of sperm per dose (500 x 106 spermatozoa). The total volume of diluted semen required was calculated by multiplying the number of does by the chosen insemination volume (0.25 ml). The amount of diluent required to dilute the ejaculate was calculated by subtracting the volume of the ejaculate from the total volume of diluted semen required (McDougall, 1995).The average time Boer and Toggenburg semen was kept from time of dilution to the beginning of insemination was approximately 7.8 and 18.6 minutes respectively. The difference in time was because the Boer semen was always collected and diluted after that of the Toggenburg had been collected and diluted.

Example of calculating dilution rates

Volume of ejaculate Swirl Density of semen 1.5 ml 5 creamy = 4000 million/ml

22

Total sperm in ejaculate No. doe doses in ejaculate Insemination volume per doe Total volume of diluted ejaculate Volume of tris required

1.5 x 4000 = 6000 million 6000 million 500 million = 12 doe doses 0.25 ml 12 doe doses x 0.25 ml = 3.0 ml 3.0 ml 1.5 ml = 1.5 ml

3.6

Cervical AI of does
Cervical AI was carried out 12 18 hours after exhibition of estrus signs, and

repeated after 8 12 hours. Cervical insemination was done using an AI pipette as described by (Pradip, 2004). Briefly, pipette was sterilized using methylated spirit and flushed with tris (diluent) prior to using it for the first time to remove any traces of the methylated spirits. A 1ml syringe was attached to the pipette with silicon rubber tubing and the pipette loaded by sucking up 0.2 ml of air followed by 0.25 ml of semen then 0.l ml of air. Many pipettes were loaded and the loaded pipettes were protected from rapid cooling by standing them in dry test tubes sitting in a water bath at 30 - 37C. Different pipettes were used for different bucks and the pipettes were changed for every 20 does. Used pipettes were immediately flushed with water and placed into a bucket of detergent. During cervical insemination the does hind legs were lifted at an angle of 60 from the ground and restrained by an assistant. A speculum lubricated with a small amount of KY jelly, was gently inserted into the vagina and the cervix visualized with the aid of light from a torch. When the cervix was not easily located or the wall of the vagina collapsed into the speculum, the doe was held more vertically. When the vagina was full of mucus, the mucus was trapped in the jaws of the speculum and withdrawn to expel the mucus. After locating the cervix, the tip of the pipette was guided into the cervical into 2-3 cervical rings. Once the pipette was in the cervix, the plunger of the syringe was pressed to expel the semen into the cervix. The speculum was held against the cervix to prevent sucking the semen out from the cervix and then gently removed. The does hind quarter was then lowered to the ground and the pipette wiped with paper towel before it was reloaded for another doe. The average time Boer and Toggenburg semen was kept from time of collection to the end of insemination was 54.4 and 65.3 minutes respectively. The difference in time was because the Boer semen was always collected after that of the Toggenburg had been collected.

23

3.7
I. II. III. IV. V. VI. VII.

Data collection and analysis

For each doe, the following data were recorded: Synchronization method Interval from implant withdraw to onset of estrus Interval from implant withdraw to 1 st AI Interval from 1st to 2 nd AI Non return to estrus at day 17 to 22 (first sign for pregnancy) Ultrasound findings at day 48 (second test for pregnancy) Males used (Toggenburg/Boer).

The data obtained was captured in Microsoft Excel and analysed using the statistical package for social scientists (SPSS) version 10.0 programs. The frequencies, percentages and chi square analysis established the conception rate of indigenous goats using fresh semen cervical artificial insemination, the comparison of estrus synchronization by use of sponges and CIDR and the conception rates of indigenous goats to Toggenburg and Boer goat semen. The chi square tests were done at 95% significance level.

24

CHAPTER FOUR RESULTS

Out of the 160 treated does, 149 (93.1%) retained the intra-vaginal device and were treated with PMSG and were given double insemination at 48 and 56 hours after the removal of the intra-vaginal device. Table 6 shows response of does to estrus synchronization treatments. More does lost CIDR inserts (n=8; 10.0 %) than sponges (n=3; 2.8%). The number of does that did not show overt estrus were more (n=3; 3.8 %) for vaginal sponges than for CIDR (n=1; 1.3%). The mean time from the intra-vaginal device withdrawal to estrus was approximately 36 hours. The percentage of does that showed overt estrus signs in sponge treatment (n=74; 96.1%) was not quite different from those treated with CIDR (n=71; 98.6%). Out of the 92 does that were diagnosed pregnant using ultrasound method, the conception rate of the does was highest 45 (48.6%) in the group aged 3 years, intermediate 29 (31.5%) in the group aged 4 years, and least 18 (19.6%) in the group aged 2 as shown in Figure 1. Age group using the chi-square test significantly (P < 0.05, X2 = 0.0001) influenced the conception rates across all the age groups.

Table 6: Response of does to estrus synchronization treatments

Treatment Number synchronised Number that lost intravaginal device Number observed in estrus Number with silent estrus Number of does inseminated Non return to estrus at day 22 Pregnant at day 48 Boer semen Sponge + CIDR + PMSG PMSG 40 40 3 (7.5%) 36 (90.0%) 1 (2.5%) 37 (92.5%) 23 (62.2%) 23 (62.2%) 4 (10.0%) 35 (87.5%) 1 (2.5%) 36 (90.0%) 24 (66.7%) 24 (66.7%) Toggenburg semen Sponge + CIDR + PMSG PMSG 40 40 0 38 (95.0%) 2 (5.0%) 40 (100%) 28 (68.3%) 27 (67.7%) 4 (10%) 36 (90.0%) 0 36 (90.0%) 18 (50.0%) 18 (50.0%) 92 (61.7%) 61.7 Total 160 11 (6.9%) 145 (90.6%) 4 (2.5%) 149 (93.1%) 93 (62.4%) Overall Percentage 100 6.9 90.6 2.5 93.1 62.4

25

60 50 Conception rate % 40 30 20 10 0 1 2 3 Age in years estimated by pairs of permanent incisor a b

Figure 1: Effect of age group on conception rate Out of the 92 does that were diagnosed pregnant using ultrasound method, the conception rate was highest 36 (75.0%), for does with body condition score of 3.5, intermediate 11 (64.7%) for does with body condition score 4.0, and least 45 (53.6%) for does with body condition score 3 (Figure 2). The chi-square test showed that nutritional status (body condition score) significantly (P < 0.05, X2 = 0.003) influenced the conception rate.
80 70 Conception rate % 60 50 40 30 20 10 0 3 3.5 Body condition score 4 a b c

Figure 2: Effect of body condition score on conception rate The overall effects of age groups and body condition score on conception rate are summarized in Table 7 and showing that body condition score of 3.5 had the highest conception rate.

26

Table 7: Relationship between age, body condition scores (BCS) and conception rate
Age (pair of permanent incisor) 2 (n = 28) 3 (n = 78) 4 (n = 43) Overall
a,b,c

Conception rate BCS 3.0 (n = 84) 9 (52.9%)

a

Overall BCS 4.0 (n = 17)

BCS 3.5 (n = 48) 9 (90.0%)

b

0 (0.0%)c 7 (70.0%)c 4 (66.7%)c 11 (64.7%)c

18 (19.6%)a 45 (48.6%)b 29 (31.5%)c 92 (61.7%)

25 (51.0%)a 11 (61.1%)a 45 (53.6%)a

13 (68.4%)b 14 (73.7%)b 36 (75.0%)b

means in the same row, with different superscripts indicate a significant difference (P <

0.05) Out of the 92 (61.7%) does which were diagnosed pregnant following cervical insemination with fresh semen, 45 (59.2%) were inseminated using Toggenburg semen while 47 (64.4%) were inseminated with Boer goat semen (Table 8). There was no significant (P > 0.05) difference in conception rates between the semen used. There was also no difference (P > 0.05) in conception rates when the does were synchronized with sponges (64.9%) or with CIDRs (58.3%).

Table 8: Effect of method of estrus synchronization and breed of buck on conception rate Conception rate
Treatment Toggenburg semen Boer semen Total Sponge + 200IU PMSG (n = 77) 27 (67.7%) 23 (62.2%) 50 (64.9%) CIDR + 200IU PMSG (n = 72) 18 (50.0%) 24 (66.7%) 42 (58.3%) Total (n = 149) 45 (59.2%) 47 (64.4%) 92 (61.7%)

Chi-square test between semen used = (P = 0.430), chi-square test between synchronization method = (P = 0.287). No significant difference between fertility of the Boer and Toggenburg. The overall fertility results are satisfactory; will increase/improve with experience.

27

CHAPTER FIVE DISCUSSION

Cervical artificial insemination using fresh semen among Uganda indigenous goats can achieve conception rate of above 60%. The current study was conducted among goats managed on unimproved pastures and no feed supplements were given. Out of the 149 inseminated goats (Table 6), 62.4% did not return to estrus 22 days post insemination. Ultrasound scanning at Day 48 post insemination, detected 61.7% does pregnant (Table 7). This figure is a satisfactory conception rate, especially when compared with conception rates obtained by some other researchers: In this respect, Armstrong (1982) achieved pregnancy rate of 60% in goats synchronized using progesterone and PMSG. Wildeus (1993) stated that 63% conception rate could be obtained when progesterone in combination with PMSG were used for synchronizing estrus in dairy goats. These figures lies within the range of 51.7 to 87.5% reported for does synchronized with intra-vaginal progestagens during the breeding and non-breeding seasons, respectively (McDougall, 1995; Freitas et al., 1996 b, Freitas et al., 1997; Greyling and Van der Nest, 2000; Motlomelo et al., 2002). The logical explanation for these variations in conception rates may be the detrimental effects of synchronization on sperm transport and survival in the female reproductive tract (Pearce and Robinson, 1985) and differences in the time of occurrence of estrous (Baril et al., 1993). In addition, low conception rates in other studies could also be a result of improper AI techniques, deficiencies in management, and changes in the environment of the does just before or after breeding (Brown and Pradip, 2007). In the present study, insemination was carried out at 12 hours after exhibition of estrus and repeated after an interval of 8 hours as described by Pradip (2004). This and proper identification of the cervix as well as penetration of at least 2 3 cervical rings or more could be responsible for the high conception rates. None of the type of estrus synchronization procedures showed any advantage over the other with respect to the conception rates. Table 8 shows that 77 and 72 does were synchronized by sponges and CIDRs, respectively, and all treated does were injected with PMSG at withdrawal. There was no significant (P > 0.05) difference in conception rates when does were synchronized with sponges or with CIDR. This is in agreement with a similar study by Ritar et al., (1990) in Cashmere does in Australia comparing CIDR with intra-vaginal sponges, and at termination of intra-vaginal treatment PMSG (200 IU). In these experiments pregnancy rates of 39% were achieved for cervical insemination. In 10

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on-farm trials with mixed-breed ewes in Minnesota, CIDR performed similarly to sponges after a 14 day treatment period and an injection of PMSG (750 IU) at the end of intravaginal treatment (Hamra et al., 1989). Conception rates were 71% CDIR and 85% for sponges, respectively. These experiments suggested no differences between sponges and CIDR devices. During the estrus synchronization, 3(2.8%) sponges and 8(10%) CIDRs were lost. According to Wildeus (2000) intra-vaginal sponges have high retention rates, greater than 90% and hence loss of 2% was within the acceptable range. The good retention rate of CIDRs in this study was another advantage for synchronizing estrus (approximately 92%) although this was slightly lower than 96% retention reported by Al-Sobayil (2006). This could have been due to improper insertion and therefore pulled out by other goats (AlSobayil, 2006), or their length not being compatible with the size of the small indigenous goats. The high rate of response to estrus synchronization using vaginal sponges (92.5%) and CIDR (88.8%) (Table 6) and the high conception rates of 64.9% and 58.3% (Table 8), respectively, further supports the benefits of estrus synchronization in goats. However, from practical perspectives, there are some factors to be considered before the use of hormonal treatments in estrus synchronization. These factors include the availability of labor at time of treatment, breeding and kidding; adequacy of kidding facilities; breed and age of the buck especially if natural breeding is used; and the cost of drug used for synchronization. Male can serve more than 50 does in the season. However, it is better to use one buck to only 10 or 15 does when an estrus protocol is used because goats show estrus at the same time (Gordon, 1997). Alternatively, breeding of the does by use of artificial insemination can be used depending on the availability of the artificial insemination service. Out of the 77 does bred using Toggenburg fresh semen 59.2% got pregnant, while out of the 72 does bred using fresh semen from Boer buck 64.4% got pregnant. There was no breed difference (P > 0.05) in conception rates as shown in Table 8. These figures were well in the range of conception rate with freshly diluted goat semen which varies from 4085% (McDougall, 1995; Pradip, 2004; Panhwar, 2007). The results indicated that the application of goat artificial insemination technology would greatly improve the utilization efficiency of fine quality bucks and promote development of goat production. Breeding activities require tremendous amounts of energy. Breeding bucks should have a satisfactory body condition score and be put on an improved plane of nutrition as the breeding approaches to enhance the quality in both viability and morphology of spermatozoa

29

produced (Teresa, 2000). In addition, the timing of the cervical AI should be right and a correct AI technique should be used (Pradip, 2004). Figure 1 and Table 7 show the influence of age on conception rates following estrus synchronization and fixed time artificial insemination. There was significant age difference (P < 0.05) between age 2 and 3, age 2 and 4, and age 3 and 4in conception rates. Age 3 had the highest while age 4 had intermediate, and age 2 had the least conception rate as shown in Figure 1. This agrees with the findings by Webb (2004) that South African indigenous does in rural communal farming systems reproductive rate increases with age and peaks at 3 to 4 years of age, remains stable and then starts to decrease. Table 7 shows that body condition score (BCS) of 3.5 following a five point score had the highest conception rate (n=36; 75.0%). There was significant body condition score difference (P < 0.05) between BCS 3 and BSC 3.5, BCS 3 and BSC 4, and BCS 3.5 and BCS 4. According to Dogan et al 2004, lactating Saanen does 2 5 years of age and with good body conditions (BCS: 2.00 to 5.00) had an overall conception rate of 51.2% at 53 days following Cervical artificial insemination (AI) with fresh diluted semen performed at a fixed time 36 and 48 hours. In the present study, AI in synchronized does was carried out at a fixed time (48 and 56 hours) following sponge and CIDR withdrawal and PMSG treatment, attained an overall conception rate of 61.7% which was higher than 51.2% achieved by Dogan et al., (2004) above. The difference in the conception rate could be due to the differences in breed, nutrition, season, use of gonadotrophins and presence of the male after sponge removal as these factors are known to influence conception rates (Greyling et al., 1997; Rosado et al., 1998; Gordon, 1999; Ungerfeld and Rubianes, 1999; Zeleke et al., 2005). This study has demonstrated that conception rate of 61.7% can be achieved in the indigenous Ugandan goats using fresh semen cervical artificial insemination. Although the study was carried out on tethered and non supplemented does, it is anticipated that better results can be obtained when improved husbandry and management practices are put into use. It should also be noted that good semen processing, semen handling, good knowledge of estrus signs, timing of the insemination and insemination techniques contribute to conception rates that can be obtained. Estrus synchronization by the use of either sponges or CIDR yielded no difference in conception rates. The use of synchronization depends among other requirements on the availability of the hormones, cost, and the need to produce uniform age groups of animals or to reduce the kidding interval. Synchronization is also useful when using artificial insemination to serve a good number of does in that it helps to reduce frequent visits and cost of the inseminator. The conception rate to Toggenburg or Boer fresh semen did not differ. This is objectively useful for farmers to develop confidence

30

in making choice of which breed to produce by use of fresh semen cervical artificial insemination. However, the conception rates were dependent on age and body condition scores of the inseminated does. It is therefore important that age and body condition score should be monitored in a well planned breeding program.

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CHAPTER SIX CONCLUSIONS AND RECOMMENDATIONS 6.1 Conclusion

- 87.5% for cervical AI using fresh semen. No significant hindrance was found and this fertility rate was satisfactory. 2. There was no significant difference in conception rates between the use of sponges and CIDRs for synchronization of indigenous goats. 3. There was no difference in conception rates of indigenous goats between using fresh semen of either Toggenburg (dairy) or Boer (meat) bucks following estrus synchronization. 4. There was significant difference in conception rates between the ages of indigenous goats inseminated. 5. There was significant difference in conception rates between the body conditions scores of the indigenous goats inseminated although body condition score of 3.5 had the highest conception rate. 6. This study indicated that cervical AI with a dose of 500 x 106 spermatozoa could be used.

1. The overall conception rate of 61.7% in this study is within the recommended range 51.7

6.2

Recommendations
There is an increasing interest in goat production in Uganda today due to increasing

market demand, pressure on the available land resource favouring production of small ruminants to large ruminants. Consequently, a number of goat crossbreeding projects are striving to improve the productivity of indigenous goats. In the implementation of these projects, more often than not, there is not enough good quality exotic or crossbred bucks to be used. This study therefore recommends that: 1. For effective use of the available quality exotic and crossbred bucks to attain more doe coverage, goat cervical AI should be adopted and incorporated in the national goat breeding projects. This technique is simple, and does not require use of expensive equipment nor cryopreservation. 2. Veterinary staff based at sub counties should be trained and equipped to collect and evaluate semen for AI extension services. Farmers in remote areas who cannot afford to own bucks can be organised into groups, and some of them trained to take part in supervision of some activities like synchronization and estrus detection.

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3. Estrus synchronization should be adopted in order to produce goats of uniform size batches for the market demand requirements. 4. National Animal Genetic Resource Centre and Data Bank at Entebbe should establish AI centre for goats for both supply of semen and training personnel, and should carry out analysis of the cost effectiveness of goat AI. 5. A more detailed study on use of frozen goat for AI of indigenous goats is required.

33

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McDonald, L. E., Pineda, M. H. (1989). Veterinary Endocrinology and Reproduction. Philadelphia, London. 10, 355 383. McDougall, I., F. (1995). Cervical artificial insemination training manual for sheep farmers. Gloucestershire, 1-17. Mgongo, F. O. K. (1988). The effect of buck teasing on synchronization of estrus in goats after intravulvo-submucosal administration of cloprostenol. Theriogenology 30: 987-995. Moore, R. W., Miller, C. M and Hall, D. R. (1988). Cervical versus laparoscopic AI of goat after PMSG injection at or 48 hours before CIDR removal. Proc. New Zealand Soc. Animal.Production. 48:6970. Morrow, D. A. (1986). Current Therapy in Theriogenology: Diagnosis, Treatment, and Prevention of Reproductive Diseases in Small & Large Animals. A Saunders Title; 2 edition, 577 632, 880 889. Motlomelo, K.C., Greyling, J.P.C., Schwalbach, L.M.J. (2002). Synchronization of oestrus in goats: the use of different progestagen treatments. Small Ruminant Research, 45, 4549. Nix, J. (2002). Tips for Heat Checking Goats. Sweetlix livestock supplement system. www.sweetlix.com. Ott, R.S., Nelson, D.R., and Hixon, J.E. 1980 a. Fertility of goats following synchronization of estrus with prostaglandin F2 . Theriogenology, 13: 341-345. Panhwar, F. (2007). Modern reproductive methods used to enhance goat production. Agricultural Research Services, United States Department of Agriculture. 1999-2008 GoatWorld.Com. Peacock, C. (1996). Improving Goat Production in the Tropics. A manual for developing workers. FARM-Africa and Oxfam (UK and Ireland. 7, 235 -252. Pearce, D.T., Robinson, T.J. (1985). Plasma progesterone concentrations, ovarian and endocrinological responses and sperm transport in ewes with synchronized oestrus. Journal of Reproduction and Fertility, 75, 4962. Pradip, G. (2004). Artificial insemination of goats; technical training manual. Nimbkar Agricultural Research Institute, Phaltan 1 - 40. Pugh, D. G. (2002). Sheep and Goat Medicine. Philadelihia Saunders. Theriogenology of sheep and goats, 144. Ritar, A. J., P. D. Ball., OMay, P. J. (1990). Artificial insemination of Cashmere goats: effects on fertility and fecundity of intra-vaginal treatment, method and time of insemination, semen freezing process, number of motile spermatozoa and age of females. Reproduction Fertility Development, 2:377384. Romano, J.E and Fernandez, A. D. (1997). Effect of service on duration of estrus and ovulation in dairy goats. Animal Reproduction Science, 47, 107112. Romano, J.E. (1998). The effect of continuous presence of bucks on hastening the onset of estrus in synchronized does during the breeding season. Small Ruminant Research, 30, 99 103.

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Romano, J.E., Crabo, B.G and Christians, C.J. (2000). Effect of sterile service on estrus duration, fertility and prolificacy in artificially inseminated dairy goats. Theriogenology, 53, 13451353. Romano, J.E. (2002). Does in proestrus-estrus hasten estrus onset in does estrous synchronized during breeding season. Applied Animal Behaviour Science, 77, 329334. Shelton, M., Lawson, J. (1982). The Effect of Season on Reproductive Activity of Meat Type Goats in Texas. U. Proceedings of the Third International Conference on Goat Production and Disease (Abst.) pp341. Singh-Knights, D and Knights, M. (2005). Feasibility of Goat Production in West Virginia- A Handbook for Beginners. West Virginia University. Page 18 19. Sohnrey, B and Holtz, W. (2005). Transcervical deep cornual insemination of goats. American Society of Animal Science 1543-1547. Teresa, W. (2000). Advanced Caprine Reproduction Methods & Techniques. Agricultural Research and Extension Programs; Langston University. 5, 20. Ungerfeld, R and Rubianes, E. (1999). Estrus response to the ram effect in Corriedale ewes primed with medroxyprogesterone during the breeding season. Small Ruminant Research, 32, 8991. Walkden-Brown, S. W and Martin, G.B. (1997). Seasonal breeding in sheep and goats: making sense of the diversity of reproductive strategies. Australia. Society of Reproduction Biology. 28, 4. Wayne, P. (2002). Introduction to Artificial Inseminating. Whitetail Heartbeat of America. www.whitetailquest.com. Webb, E. C., Mamabolo, M. J. (2004). Production and reproduction characteristics of South African indigenous goats in communal farming systems. South African Journal of Animal Science 2004, 34 (Supplement 1) South African Society for Animal Science Peer-reviewed paper: 8th International Conference on Goats, 237. Whitley, N. C., Jackson, D.J. (2004). An update on estrus synchronization in goats: a minor species. Journal of Animal Science, E-Supplement: E270-276. Wildeus, S. (1993). Techniques to improve reproductive efficiency in goats and sheep. Proceedings of the 1993 Symposium on the Health and Diseases of Small Ruminants Jackson Hole, Wyoming, USA, June 12-13, 1993, 130-142. Wildeus, S. (2000). Current concepts in synchronization of estrus: Sheep and goats. Agricultural Research Station, Virginia State University, Petersburg 23806, Proceedings of the American Society of Animal Science, 1999. 1 14. William, F and Braun Jr. (1985). Manipulation of the reproductive cycle of the doe. Proceedingsof The 1985 symposium of American Association of Sheep and Goats Practitioners, February 27-28, 1985, 77- 85. Youngquist, R. S. (1997). Current therapy in large animal Theriogenology. W. B. Sanders campany USA, 62, 483.

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Zeleke, M., Greyling, J.P.C., Schwalbach, L.M.J., Muller, T., Erasmus, J.A. (2005). Effect of progestagen and PMSG on oestrous synchronization and fertility in Dorper ewes during the transition period. Small Ruminant Research, 56, 4753.

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APPENDICES APPENDIX I Map of Kamuli showing goat AI research area

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APPENDIX II List of equipment required For semen collection

Artificial vagina Liners Sperm glasses Insulating sleeve for sperm glass Box of thick rubber bands for holding liner to AV Litre of industrial methylated spirit for sterilizing liners Kettle to heat water for AV Tube of KY jerry to lubricate AV Hand held pump action sprayer to dispense methylated spirit Paper towels

For semen dilution

Bowl or water bath filled with water at 30oC Test tube racks Disposable test tubes or glass test tubes Box of Pasteur pipettes with rubber teat to transfer semen from sperm glass to test tube 1ml syringes for measuring UHT skim milk diluent into semen 250ml beaker to place test tube of diluted semen into to allow cool to 16oC Thermometers to check temperature of water for AV and water for holding and cooling semen Record sheet to record ejaculate details and calculate dilution rate

For insemination
AI pipette with 1ml syringe attached, to measure and draw up semen dose or goat AI gun 0.25ml straws AI sheath Container of PVA powder to seal straws Scissors to cut straws Roll of paper to dry straws

40

Speculum Flash light and batteries Tube of KY to lubricate speculum Roll of paper towels to wipe AI pipette between inseminations People to restrain does for insemination

For cleaning equipment

Laboratory detergent Test tube brash for cleaning sperm glasses Deionised water for rinsing sperm glasses Roll of house hold foil to cover sperm glasses while being sterilized Access to a clean oven to sterilize sperm glasses Industrial/surgical methylated spirit

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APPENDIX III Table 10: Data collection table

Doe mumber Age Body condition score Pd scan - pre AI Pregnant Sponge synchronized CIDR synchronized Sponge/CIDRwithdrawal time PMSG injection time Estrous time Interval withdrawalestrous Toggenburg buck semen used Boer buck semen used First AI time Interval withdrawal first IA time Second AI time Interval first second AI time Pd non return to estrus pregnant Pd scan post AI pregnant-CR Sponge lost CIDR lost Data collection table

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