SD9700017 CONCEPTION RATES FOLLOWING OESTRUS ...

SD9700017

CONCEPTION RATES FOLLOWING OESTRUSSYNCHRONIZATION AND ARTIFICIAL

INSEMINATION IN THE NUBIAN GOATS

BY

AMBROSE SAMUEL JUBARAB.V.Sc. Assiut University (Egypt)

(May 1986)

A Thesis Submitted in Partial Fulfilment or theRequirements for the Degree of M.Sc.

Tropical Animal Production

Department of Animal Husbandry and GeneticsFaculty of Animal Production

University of Khartoum

-07July, 1996

We regret thatsome of the pagesin this report may

not be up to theproper legibilitystandards, eventhough the best

possible copy wasused for scanning

Dedication

To the Memory of my Parents, Samuel Jubara, Ludia Lurit.

To those who persevere my absency for these couple of years; my wifeLucy and Daughters Delight and Sophie.

(i)

TABL: OF CONTENTS

£a&e

Acknowledgement (vii)

Abstract (viii)

Abstract (Arabic) (x)

List ol'Tables (v)

List of Figures (vi)

INTRODUCTION 1

1.0 Literature Review 31.1 Economic Importance c 1 Nubian Goats 3

A. Description of Nubian Goats • 3B. Population Size 3C. Distribution 3D. Production Potentials 4

(i) Milk Production 4(ii) Meat Production 4(iii)Growth Rales of Nubian Kids 4

I',. Cuiiliibiition nl Nubian Goals in llicNational Economy 5

I1. lil'loils Coiuludal lor Cicuclic Improvementof Nubian Goats 6

1.3 Oestrus, Oestrus Period, Oestrous Cycle 6(i) Oestrus 6(ii) Oestrous Cycle Length 6(iii) Duration of Oestrus Period 7

1.4 Signs of Oestrus 71.5 Hormonal Events During Oestrus 7

(i) Gonadotrophins and Oestrogen 8(ii) Progesterone and The Corpus Luteum 9(iii) Prostaglandins and Luteolysis 9

1.6 Oestrus in Pregnancy 101.7 Factors Affecting Oest ous Cycle 10

(i) Climatic Conditions H(ii) Nutrition U

(ii)

1.8 Artificia](Hormonal) Control of Oestrous 11A. Synchronization of Oestrus Using

Prostaglandin and its Synthetic Analogues 11(i) Dose Levels of Prostaglandin (PGF2oc) 12(ii) Responsive Periods of Corpus Luteum to

Prostaglandin and its Analogues Treatment. .... 12(iii) Interval Between Prostaglandin Doses. 13(iv) Interval from Prostaglandin Injection

to Oestrus. 13(v) Fertility Due to T-eatment with Prostaglandin

or its Analogues. 13B. Synchronization of Oestious Using Progesterone

Impregnated Sponges. 13C. Synchronization of Oestr.is Using Progesterone/

Progestagen and Pregna~.t Mare Serum Gonadotrophin(PMSG). 14

1.9 Non-hormonal Methods of Oestrus Synchronization. .... 15A. Presence of the Male 15B. Factors Involved in the Teasing Effects 16

(i) Introduction of the Male. 16(ii) Continuous Exposure to Male. 17

1.10 Artificial Insemination (A.I) and Hand Mating 17A. Advantages of A.I. and Conception Rates Due to

A.I. in Goats. 17B. Methods of Semen Collection. 17

(i) Artificial Vagina (A.V). 17(ii) Electro-ejaculation. 18

C. Evaluation of Semen. 18D. Semen Diluents. ISE. Semen Dilution. 19I1'. liiNeiiiin.ilioii Tcdini(|iii\s. W

(i) Timing of one Vs. two Inseminations. 19(ii) Sites of Semen Deposition. 20(iii) Insemination Procedure. . . 20

G. Factors Affecting Fertility at A.I. 211.11 Hand Mating Technique. 211.12 Pregnancy Diagnosis (I'D). 21

I. Gestation Period. 21II. Importance of Pregnancy Diagnosis. 21

III. Methods of Pregnancy Diagnosis. 22(i) Palpation Techniques. • 22a. Abdominal PalpUion (Ballotement). 22b. Bimanual Rectal Probe-Abdominal

Palpation Technique. 22c. Cervical Palpation. 22

(ii) Pregnancy Diagnosis by Hormonal Assay 22

(iii)

A. Hormones of Gestation. 22a. Progesterone. 22b. Prostaglandin (PGF2oc) and Oxytocin 23c. Estrone Sulfate. 23d. Testosterone. 23e. Oestrac!iol-17j8. 24

B. Hormonal Assay Methods for Pregnancy Diagnosis. 24(iii)Ultrasonic Techniques for PD. 25a. Amplitude Depth Ultrasonic (A-Mode Technique) 25b. The Doppler Ultrasound Technique. 25c. Real-Time (B-Mode) Ultrasound Scanning. 25

(iv) Radiographic Technique. 26(v) Udder iixainination. 26(vi) Other Methods of Pregnancy Diagnosis 26a. Corpus Luteum Biometry and Weight. 26b. Vaginal Cytology. 26c. Absence of Return to Oestrus. 26

CHAPTER TWO:

2.0 Materials and Methods. 272.1 Study Area. 272.2 Environmental Condition. 272.3 Experimental Animals. 272.4 Husbandry. 28

2.4.1 Aging. 282.4.2 Housing. 282.4.3 Health. : 302.4.4 Foaling. 30

2.5 Hxpcrimcnlalion. 312.5.1 Adaptation Period. 312.5.2 Treatment. 322.5.3 Artificial Insemination and Hand Mating 37

A. Semen Collection. 37B. Semen Evaluation. 37

(i) Colour and Volume. 37(ii) Mass Activity. 37

C. Concentration of Spermatozoa. 38D. Semen Dilution. 39

(iii) Insemination Techniques. 392.5.4 Return to Service. 402.5.5 Conception Rates. 422.5.6 Pregnancy Diagnosis (PD). 42

1. Pregnancy Diagnosis by Progesterone Assay 42Using Radio-immuno Assay (RIA) 42

(iv)

Page

II. Pregnancy Diagnosis by Abdominal PalpationTechnique. 45

2.6 Statistical Analysis. 45

CHAPTER THREE:

3.0 Results. 463.1 Oestrus Parameters. 46

3.1.1 Response of Does to Different Treatments. 463.1.2 Insertion, Retention and Removal of

Progesterone Impregnated Intra-vaginalSponges. 46

3.1.3 Interval from the end of Treatment end to Oestrus. ... 463.1.4 Duration of Oestrus Period. 473.1.5 Frequencies of Manifestation of Oestrus Signs 47

3.2 Fertility Measures. 583.2.1 Return to Oestrus Rates. 5$3.2.2 Conception Rates. 583.2.3 Pregnancy Rates. 593.2.4 pseudo-Pregnaroy. 623.2.5 Oestrus in Pregnacy. 62

CHAPTER FOUR:

Discussion. 73Conclusions. 80Recommendations. 81List of RtTi'rences. 82

(v)

LIST OF TABLES

Page

Table 1: Environmental Conditions During Experimental Period(July - December). 28

Table 2: Ingredients and Composition of Ration 1 and 2 Fed

During the Experiment. 31

Table 3: Scoring System for Mass Activity (Wave Motion). 39

Table 4: Number of Does Retainii g the Sponges and the Easiness

of insertion and removal. 47Table 5: Does Responded to Treatments and Mean interval from

Treatment end to onset of Oestrus (hours) and MeanDuration of Oestrus period (hours) 48

Table 6: Relative Frequency (R.F) of Incidence of OestrusBehaviour in Relation to onset of Oestrus (Time 0)in hours. 57

Table 7: Number of Docs Inseminated, Number Returning toOestrus, Conception Rates and Pregnancy Rates andDoes Exhibiting Oestrus in Pregnancy. 60

Table 8: Serum Progesterone Co.icentration of the SudaneseNubian Goats 10 Weeks Post Insemination. 62

(vi)

LIST OF FIGURES AND PLATES

Plate No. 1: Physical Characteristics of Sudanese NubianGoats.

Plate No. 2: Pens for Intensive Housing of Nubian Goats.

Plate No. 3: Equipment for A.I. and Application ofIntravaginal Sponge from Left to Right Straw, Gun,Plastic Catheter, Vaginal Speculum, Light Stick,Artificial Vagina, Semen Collecting Tube (Graduated),Stringed Sponge, Plunger, Bevelled end Tube.

Plate No. 4: Restraining Doe for Sponge Application and A.I. 35

Plate No. 5: Insemination Procedure. 41

Plate No. 6: Inlravaginal Application of the ImpregnatedSponge. 36

Fig. No. 7: Frequencies of Maiiii'cslalion ul Signs ofOestrus.

Fig. No. K: Standard Curve. 43

Fig. No. 9: Graphical Presentation of Progesterone Profilesin Serum of Pregnant, Non-pregnant Goats Post-in.scmiiulioM. 63-72

(vii)

ACKNOWLEDGEMENTS

To my Supervisor Dr. Muna Mahgoub M. A; I extend my sincere gratitude forher encouragement and guidance throughout the year. I would like to register myfaithful bow to the blessed spirit and the message within her that does not ignore thereality of life which strengthen me to overcome feelings of - helplesseness andhopelessness. ?.

This work was initiated by Dr. Sharaf El Din Makawi, Head Department ofSurgery Gynaecology and Obstetrics who without reservations and open heartedlysurrounded me with his capable technical advice and guidance and genuinely markedand bloomed this thesis; to him I am extremely grateful.

The experimental animals, equipment and other working facilities in the goatsup grading project, Directorate of Animal Resources - Khartoum State were madeavailable for me by Dr. Muna Awad, who with the entire staff of the projectsurrrounded me witli cordial friendly working atmosphere, to them I extend my sinceregratitude.

Worthly acknowledged is the Administration and staff of Radio IsotopeLaboratory - Souba for provision of the progesterone Kits and to my colleagues Dr.Mohsin and Dr. Faisal of the same Ur:t who voluntarily trained me though for shortbut satisfactory and dutifully processec my sample when I was haridicaped, to them Iam greatly indebted.

My thanks are also due to Mr. Ahmed Abdel Gadier of Juba University, Mrs.MasUua Ibrahim, Miss Yousra A. of Federal A.I. Centre for their technical assistanceand lo Gabish Veterinary Clinic for allowing me to process my samples in theirlaboratory.

I am greatly indebted to colleagues Dr. I. I. Julia, Charles, E. W. for provisionof film and photography and Dr. John, K. who aspired me his time during samplescollection, and to my colleagues in batch five, my room mates wru^ contributed in oneway or another to make this work through. The moral and financial support of mybrother Major (ARMY) Justin Ladu was very much appreciated.

I was granted study leave with pay by my employer State Government of BahrEl Jebel, I am very grateful for that. Very much acknowledge is,the sacrifice madeby my mother-in-law in caring for my children during my absence.

Lastly, I gratefully acknowledge the effort exerted by Mr. Sylvanus YamaLumata, of Humanitarian Aid Commission, Early Warning Unit, Khartoum, fm Tsetting and typing this manuscript in su:h an excellent form.

(viii)

ABSTRACT v

This Experiment was designed to investigate into the efficacy of different

hormonal treatments in inducing and synchronizing oestrus in Sudanese Nubian goats

and their fertility following a fixed time artificial insemination programme using

Safcnen buck semen.

From a flock of 150 females of mixed breeds and crosses, 34 female Nubian

goats were selected and grouped by ages such that each age1 group should be

represented under each treatment. They were then randomly assigned to different

treatments Viz:

Four females were allowed to cycle naturally as control (Treatment A): ten were

injected intramuscularly with 125^g Cloprostenol, two doses given 13 days apart

(Treatment B): other ten females were fitted intravaginally with sponges impregnated

with 40 mg progesterone for 16 days (Treatment C): the last ten females were treated

as C but were injected intramuscularly with "300 I:U pregnant Mare Serum

Gonadotrophin (PMSG) two days before removal of sponges (Treatment D): one

vascctomizcd buck and other aspermic were immediately introduced to the treated

groups to aid in detection and initiation of oestrus. :

Dose in each treatment were monitored for interval from treatments to onset of

oestrus, oestrus signs, duration of oestrus period. They were then inseminated at a

fixed time seen in oestrus or not, meanwhile the controlled group were handmated.

Conception rates were estimated as non return rates, and early pregnancy (3 - 10

weeks) after insemination was diagnosed by prcgesterone Radio-Immuno-Assay (RIA)

and late pregnancy (90 - 110 days) post-insemination was diagnosed by abdominal

palpation technique.

The obtained results indicated that all the employed treatments were capable of

inducing and synchronizing oestrus in Sudanese Nubian goatSi Treatment B being

significantly higher than other treatments (P < 0.05).

(ix)

There was significant difference between treatments (P < 0.05) as far as the

duration of oestrus period is concerned in this study. Pregnancy rates were

significantly different between treatments (P < 0.05) in both RAI and abdominal

palpation methods of pregnancy diagnosis. However, the overall pregnancy rate by

RIA (12 = 37.50%) was less than that diagnosed by abdominal palpation (22 = 64%).

However, kidding confirmed the accuracy of diagnosis by RIA.

Treatment B has more advantages than C and D in oestrus induction and

synchronization and could easily be applied in a large flock of different ages with

minimal labour required, while C and D proved to be difficult in its application in non-

parous goats and requires assistance and some hygienic measures during application.

This study recommended cloprosterol (Treatment B), fixed time artificial

insemination, progesterone RIA for monitoring early pregnancy and return to heat, and

improve nutritional programs as strong pillars for adoption of intensive system for

dairy goats husbandry in the major tov'ns and rural areas during cropping season.

Further research in needed using progesterone impregnated sponges with

different concentrations and monitoring the absorped amount of progesterone into

blood circulation by RIA, before its recommendation for field application.

(x)

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INTRODUCTION

- 1 -

INTRODUCTION

Nubian goats are owned in Urban or peri-urban production systems which are

assuming increasing importance in the Sudan. They roam freely during the day,

scavenging in towns feeding on mixed natural vegetation, crops by-products, papers

and other garbages, destroying sensitive ecosystem, and are confined and fed

household wastes and occasionally concentrates at night. They are also owned by

settled agriculturalists where they are tethered during cropping season to prevent crops

destruction.

Areas of distribution of Nubian {joats in the Sudan, lie in the arid, semi-arid

zone where natural grazing lands are scarce and goats therefore are kept under sub-

optimal management. The trend here th :refore necessiates adoption of intensive type

of management which is in turn surrounded with controversial account on goat in that

it prevent the destructive effects of goats; and on the other hand it decreased time

available for observation of an individual goat for signs of oestrus in the absence of

male and hence will affect productivity of the goat. As such, synchronization of

oestrus as an aid for controlled breeding techniques removes the, need for oestrus

detection. This technique now is crucial as there is a growing jnterest among the

urban population in rearing high yielding expensive exotic dairy goatjS and their crosses

for family daily need of milk.

Synchronization of oestrus in farm animals arc means for bringing groups of

female animals into heal together in response to some hormonal treatments. These

animals will then conceive and give birch at almost similar times. The importance of

synchronization of oestrus in goats is managerial. It allows a farmer to predictJhe

time of oestrus and ovulation with reasonable accuracy (Smith, -4980; Britt, 1984).

This will reduce the time required for detection of oestrus signs and may make it

possible to breed at a fixed time without he need to detect oestrus signs (Hafez, 1987).

Other benefits of synchronizing oestrus include feeding animals in uniform group with

a diet according to the stage of production, it also facilitates supervision of a group of

kidding does, thus reduces neonatal mortality, it helps as well in organizing weaning,

— f —

fattening and marketing a batch of kids and rationalizes the use of labour,-buildings

and oilier resources, and secure continuous supply of milk and kids.

Several hormones /now being used in the induction and synchronization of

oestrus in goats; among which are prostaglandin F2« (PGF2 oc) and its synthetic

analogues Estrumate or Cloprosteno1, and synthetic progesterone (progestagens)

impregnated sponges such as Veramic and Cronolone. However, in the Sudan, no

work has been so far reported concerning induction and synchronization of oestrus in

indigenous Nubian goats using progesterone impregnated sponges, nor with PGF2«

or one of its synthetic analogue.

Despite the sizable contribution of the Nubian goats to the national economy,

little efforts were undertaken to improve their reproductive performance and hence

productivity. Futhermore, nothing has been known about. the reproductive

performance of the Nubian goats under hormonally controlled breeding condition in

the Sudan. Therefore, this work was undertaken to explore the following objectives:

1. Testing the efficacy of different hormonal methods in inducing and

synchronizing oestrus in indigenous Nubian goats.

2. Assessing ihcir fertility following synchronization of oestrus and a fixed time

artificial insemination.

- 3 -

1. LITERATURE REVIEW

1.1 Economic Importance of The Nubian Goat

i A. Description of Nubian Qoat

I The Nubian goat, among other indigenous goats (Desert goat, Nilotic dwarf,

f Taggari) is the only acknowledged dairy goat in the Sudan (Hassan and El Derani,

~" 1990) and is among the best dairy goafs in Africa (Devendra and McLeaory, 1982).

£

s • Nubian proper is the name for cistinction of Sudanese Nubian goats from other.v

,f similar breeds (FAO, 1991), its physical characteristics were described by Mason,

I ; , (1967); Devendra and McLeory, (1982); El Nairn, (1992), as a large size goat of 70 -

£ - 75 cm height males weighing 50 - 70 Kg and females 40 - 60 Kg. The head is small

y to medium, the forehead prominent with markedly convex profile in males and females

\l' and depressed just behind nostrils. Horns when present are rather light and of medium

*7 length; simple or partially twisted backwards or divergent sweep in males and usually

*4 backwards sweeping in females. Ears are long (25 cm), broad and pendulous,

% typically lop with bottom one-third turning upwards, white spottedwith black colour.

& Toggles, short in length, occur occasionally in both sexes. Beard usually absent. Legsif'p / are long but well proportioned. Colour generally black, other colours from dark

jf chocolate brown, light fawn also occur. Longer hairs on front legs and especially on

|S hind quarters and hind legs give apperance of breeches. (Plate No, 1).tt Z

t_k t K. Population Si/.ey , _e Nubian goats constitute 2.5 million out of 18.7 million head of goat in die:V\S Sudan (FAO, 1992; AOAD, 1994).

'"^j?- C. Distribution

:1t Distribution of Nubian goats throughout the arid and extreme arid areas

ffj .' essentially along rivers and in urban or peri-urban areas of the Northern part of the

- - Sudan North of 12°N and Westwards to El Obeid (FAO, 1991); •

r

- 4 -

D. Production Potentials

(i) Milk Production:

The milk yield and lactational lergdi of the Sudanese Nubian goat were 73.5

Kg, 147 days respectively with 1.5 - 2.0 Kg/day (Sulieman and El Shafei, 1984) and

were 7i.S±80 litres, 121 ±47 days with 0.6 litre/day (Khalaffalla, and Suliernan,

1990). These values were less than hose reported in temperate breeds, where it

reached 249.3±86 litres in a lactational length of 253±56 days in Saanen breed and

244.3±80 litres over a lactational length of 206±71 day*in Toggenburg breed and

162.4 + 73 litres over a lactational length of 173.5 ±49 days in Anglo-Nubian breed

(Khalaffalla and Sulieman, 1990). These values were within the .range reported for

local goats in other tropics by Devendra and Burns (1970), which were 160 Kg over

a lactation period of 120 days in Boer goats and average yield of 180 Kg over a 180

days lactation period in Malabar goat and those reported by El Nairn (1979) and

Kudouda (1985) in Sudan (73.5 - 110 Kg over a lactation length of 90 - 147 days) in

Nubian goat. Percent of lactating females Nubian goat increases above 70% in January

and fall to 40% in April Uien begins to increase to 60% in Nqvember to peak in

January again (FAO, 1991). ;

(ii) Meat Production: ,

1:1 Taycb, et al. (1987) reported < dressing percentage of ,43.2 at 14.1 Kg live

weigh! of Nubian goat fed with cotton seed cake as nitrogen source and a dressing

percentage of 38.4 at 12.1 Kg live weight with blood as nitrogen, sou ice. The carcass

composed of 23.0 and 20.7 percent gutlill in the previously mentioned two samples.

Head is 9.9 and 11 percent, hide 8.9 and 9.2 percent, feet 4.5 and 4.4 percent,

lean/bone/fat percent at 52.2/28.1/19.7 percent in cotton seed.cake supplemented

animals and 45.4/28.8/25.8 in blood supplemented ones. •

(iii) Growth Rates of Nubian Kids:

Birth weight of Nubian kids were reported by Maglad and Kudouda, (1987) to

be 2.96±0.07 Kg (males 2.5±0.5, females 2.1 ±0.3). Males were found to be

heavier than females at birth and upto 7 months of age. At 12 months males weighed

22.2 Kg and females 18.0 Kg. Average daily again at 1 - 4 weeks was 60 gm for

males, 70 gm for females and at 5 - 10 weeks males 60 gm, females 50 gm and at 21-

28 weeks males 90 gm, females 60 gm, and from birth to 1 year, males 56 gm and

females 50 gm.

E. Contribution..of Nubian.Goat in The National Economy ,

The role of goats rearing for the human population can most clearly be seen

from the ratio of goats to people. In Sudan, more than one goat is kept per two

inhabitants (Peters and Hoist, 1981). Nubian goats in the Sudan are kept to provide

human with animal protein as well as possibilities for earnings especially in times of

economic crises or under natural conditions which limit the rearing of larger

ruminants. Nubian goats contribution to the country's needs of milk and meat were

estimated to be 23.3%, 5% respectively (AOAD, 1982).

F. Efforts Conducted for Genetic Inprovement of Nubian Goats

Although milk production is a moderately heritable trait, it would take time to

achieve substantial increase by selection within the existing genotypes, because the

initial level of production is so low an-.i therefore more rapid increases might be

expected from cross breeding with exotic genotype (Kassahun, et al. 1989).

Exotic pure breeds of goats (Saanen, Toggenburg, Anglo-Nubian) were

introduced into the Sudan in 1976 as part of llic technical aid of the Overseas

Development Administration (ODA) of the Uniled Kingdom (KhalalTalla and El Shafei,

1990) for improving milk and meat production through cross-breeding with local

Nubian goats and multiplication of the pure breeds. Available information from 50%

crossbred betwee Nubian goat x Anglo-Nubian showed that lactational milk yield and

lactational length of crossbred does were 143.4±93 litres and 142±43 days

respectively, which was higher than that of local goats, and the average daily milk

yield almost equal to that scored by the temperate breed (i.O litre) and higher than 0.6

litre attained by the indigenous goat (Khalaffalla and Sulieman, 1990). El Nairn,

- 6 -

(1992) reported some results of 75% crossbred goat between Nubian x Toggenburg

with an average milk yield of 7 lbs per day, and a crossbred between Nubian x British

Alphine that gave 6 lbs per day, Nubian x Damascus x Toggenburg that gave 6 IBs

per day, Nubian x Saanen x Toggenburg that gave 7.5 lbs per day.-

1-2 Oestrus, Oestrus Period. Oestrus Cycle

(i) Oestrus:

Oestrus is a periodical sexual excitement in females brought about by the

ovarian activities under the control of anterior pituitary gland. (Devendra and

McLeory, 1982). Or it is a physiological state in which female animals seek males

and stand to be mounted and served. (Mackenzie, 1967; Lindsay and Fletcher, 1972;

Laing, 1979). Onset of oestrus was deemed to have occurred when the doe was served

by the buck (Llewelyn and Ogaa, 1995).

(ii) Oestrus Cycle Length: , ,

Tropical goats are reported to undergo ovarian activity throughout the year

(Devendra and Burns, 1983). The average length of oestrus cycle of goats in the

tropics is 18 - 21 days (Devendra and McLeory, 1982). In the Sudanese Nubian goat,

the length of oestrus cycle ranges from 19 - 24 days (Kudouda, 1985). Normal cycles

of <30 days and extended cycles of >30 days were reported by Llewelyn and Ogaa

(1995) in indigenous goals in Zimbabwe. Temperate broods also showed length of

oestrus cycle of 22.8, 23, 16.6 days in dry does, milking does .and yearling kids

respectively of both Saanen and Toggenburg in USA (Devendra and McLeory, 1982).

Short cycles of 5 - 10 days in Anglo-Nubian goats in philiphines^and 2 - 1 2 days in

Saanen and Toggenburg in USA were reported by the previous authors.

(iii) Duration of Oestrus Period:

Duration of Oestrus period is estii laled as the interval between first and last

coitus (Llewelyn and Ogaa, 1995). Mean duration for Oestrus periods in the tropics

range from 17 - 48 hours (Devendra and Burns, 1983). It was 12 - 48 hours in the

Sudanese Nubian goats (Kudouda, 1985). »

- 7 -

1.3 Signs of Oestrus

Mackenzie, (1967) reported that, female goats in oestrus are restless, bleating,

wagging tails, with red swollen vulva often showing discharges and the doe will stand

to be mounted by a buck or other females or it may mount other females. These signs

were also reported by Smith, (1980) in addition to the frequent urination of the female

and depressed appetite. The latter sign however, was not common in the Sudanese

Nubian nannies (Abdel Aziz; et al, 1982). Tail wagging was the most reliable

indicator of oestrus of the various behavioural traits (Llewelyn; et al, 1993) and its

onset was closely associated with the onset of the trait of seeking out a buck. In

contrast, bleating and restlessness were less pronounced and occurred only in 50 - 60%

of the oestrus periods. Incidence of bleating during oestrus was higher in the absence

of a buck (Coe and Allrich, 1989). Does in oestrus do mount other does, but the

frequency of this trait was low and was usually directed towards other does in oestrus;

as such, mounting behaviour was suggested to be an expression of dominance by the

high ranking females (Mathews, 1989). , .

Discharge of a thin clear mucus from vulva on oestrus day which became

cloudy, and thick creamy afterwards, was atributed to the cytologjcal changes in the

vaginal epithelium due to oestrogenic activity (Pretorius, 1977), which may sometimes

allows discharge to persist up to 7 days. This could be due to oestrogen secretion by

a new wave of developing follicles, and may explain the renewed interest shown by

the male on day 3, (Llewelyn; et al, 1993). Does were not receptive on day 3,

however possibly due to refractoriness to oestrogen (Carrick and Shelton, 19o9), or

to the antagonistic effects of progesterone (Davidge; et al, 1987); • The incidence of

vulval swelling was maximal on oestrus day and subsided 2 - 3 days later. Vulval

swelling and mucus discharge are good indicator of impending oestrus (Llewelyn,

1993).

1-4 Hormonal Events During Oestrus

(i) Gonadotrophins and Oestrogen: ;.

Oesirous behaviour is fully expressed when oestradiol concentration reached

threshold necessary to elicit positive feedback on lutenizing hormone (LH) secretion

t v

- 8 -

luteolysis, and the fall in plasma progesterone concentrations also lead to increased LH

frequency pulse from basal level as reported by Sutherland (1987). This suggests that

progesterone also regulates LH frequency pulse in goat as in sheep (Karsch, et al,

1983). During oestrus and until formation of a new corpusluteum after ovulation, the

progesterone concentration is reduced to a negligible value of 0.2 ng/ml. This amount

was believed to be of adrenal origin (Robertson, 1977). However, in sheep, no

preovulatory progesterone is secreted from the follicle as in other mammalian species.

Lleweyln, et al,; (1993) found that, the frequency of LH pulses remained elevated

although progesterone concentrations had risen to 2 ng/ml. This suggests that the

frequency of LH pulse is not modified in goat until progesterone concentrations rise

to more than 2-ng/ml (Sutherland, \lW: Clicmincau, ct al, 1988). Similar pattern was

seen on d;iy 3 o( the cycle in cow (Walters, ct al, 1084) and corresponds to a new

wave of follicular development (Savio, et al, 1990).

Mean peak of LH concentraiions was reported to be 102.1 ±7.8 ng/ml by

Chemineau, et al, (1982) with a surge lasted around 10 hours. Night, et al, (1990)

reported peak LH heights from 54 lo 114 ng/ml, m'mg ovine reference standards,

whereas Bretelaff, et al, (1988) obtained peak LH values above 300 ng/ml using

bovine LH as reference. In the goat, ovulation was found to occur 20 - 26 hours after

the LH surge (Mori and Kano. 1984; Greyling and van Niekerk, 1990 a) and

approximately 36 - 42 hours after onset of oestrus in British white docs.

(ii) Progesterone and The Corpus Luteum:

Corpus luteum attains full secretory activity of progesterone by about day 6 to

day 8 of die oestrous cycle and continues secreting at fairly constant level until day 15

(Edgar and Ronaldson, 1958). Maximum concentrations of progesterone being reached

on day 8 and then begin to fall i. day or two days before the next oestrous

(Cunningham, et al, 1975; Quirke. et al, 1979). Maximum concentrations of

progesterone during luteal phase was 2 ng/ml (Stabenfeldt, et al, 1969). Season and

nutrition (Lamond, ct al, 1972) as v\*.il as breed and ovulation rate (Thorburn, et al,

1969); Bindon, ct al, 1975; Quirke and Gosling, 1976) can influence the maximum

concentration of progesterone. Administration of exogenous progesterone in die early

days of the oestrous cycle, markedly affectd the development of corpus luteum and

may span its activity (Tlnvaites, 1971). This may be due to the interference of

progesterone with the normal mechanism responsible for the normal establishment of

luteal tissues.

(iii) Prostaglandins And Luteolysis:

Progesterone stimulates'the endonietrium to synthesize and store the luteolysis

agent (Wilson, et al, 1972). Pros tag land in (PGF2oc) is released from the uterus to

cause regression of the corpus luteum (Coding, 1974). What signals die rapid release

of PGF2oc at the end of the cycle? Alwachi, et al, (1979) evidenced a local control

exerted by die ovary on PGF2 oc synthesis in the endometrium of adjacent uterine horn

possibly via the involvement of oestrogen. Therefore administration of exogenous

oestradiol in appropriate doses near the end of tlie cycle will induce luteolysis as the

endogenous production of oestradial begins about 48 hours before the onset of oestrus

(Bjersing, et al, 1972).

1-5 Oestrus in Pregnancy

Symptoms of both physiological and psychological heat during pregnancy have

been noted long ago in several mammalian species such as cpws(Donald, 1943);

elephanis(Perry, 1953); ewes(William, 1956). Such heats which are not accompanied

by ovulation, arc not confined (o any pail of the gestation period and may occur

shortly before parturition in sheep and goals (William, el al. 1-950; Btclmoi, el al,

1974).

Pregnant ewes experiencing oestrus in early pregnancy can reach 13.7%

(Younis and Afifi, 1979); a value of 21% and 62% was reported by William, et al,

(1956). The proportion of oestrus in early pregnancy was lower; in younger females

of 1.5 - 2.5 years man older ones according to the former authors.

The most probable reasons for occurrence of oestrus during early gestation may

be due to excessive feeding on green lucerne during mating and early gestation as

lucerne is known to contain some oestrogenic compounds (Shutt, 1976). It may also

ft? - 10 -

4% lucerne is known to contain some oestrogenic compounds (Shutt, 1976). It may also

?/ be due to inherited endocrine activity such as individual physiological difference in the

£\], activity of pituitary gland resulting into variable amount of oestrogen in the blood

^ 1 sufficient to bring heat inspite of the depressing effect of the progesterone (Mirskaria

and Smirnov, 1941). Some authors were reported by Gordon, (1983) to have gone so

*j* far as to suggest that the frequent occurrence of oestrus in the pregnant ewe, make the

,T T practice of putting rams with ewes to detect non-pregnant females questionable.

jr 1-6 Factors Affecting Oestrous Cycte

T % Apart from diseases and hormoi>es, there are other factors that exert a

considerable influence on oestrous cycle cither directly or indirectly (Williamson and

Payne, 1978). The most important o" these factors appears to be the plane of

~~- nutrition, climatic conditions and housing.

~ ' (i) Climatic Conditions: ,

TS; The duration of oestrus period in West African dwarf goat was found to be

;i " longer in early dry and late dry season (36.1 hours and 32.8 hours .respectively, than

'/ in the early rainy season 19.8 hours) and late rainy season 22.4( hours (AJcusu and

*. i- Egbunike, 1990). When the fore-mentioned animals were treated with gonadotrophins,

-*V" 50% of the goats showed a positive response during the rainy months compared with

i; less lhan 35% of the goals treated during the dry months (Rosnina, et al 1992),

Oeslrous cycle length was found to be 21.2, 20.7, 19.8 days and the duration of

oestrus period 38.4, 40.8, 49.1 hours in Winter, Summer and Aulum respectively in

Egyptian Baladi goats under semi-arid environment (Younis, et al 1989).

£rr. Season and housing were found tD influence ovarian activities hence fertility,

possibly due to changes in temperature and photoperiod. The author (Llewelyn and

Ogaa, 1995) observed that the proportion of normal cycles occurring in winter was

^ 87.5% and 77.7% for goats in single ar j group pens respectively, falling to 62.5%

^ and 37.5% respectively in spring. Goats housed communally, experienced a greater

j . | fall in percentage of normal cycles in spring, possibly due ;to increased stress

associated with group feeding, and that oestrus duration in winter was 1.94 days VS

- 1 1 -

1.13 days in spring and the luteal phase length was not affected by season, but was

significantly associated with housing (16.93 days in single pens VS 18.32 days in

group pens).

(ii) Nutrition:

Onset of oestrous cycle, and attainrr ^nt of puberty and sexual maturity in goats

were very much delayed in inadequately Ted female goals (Williamson and Payne,

1978). Under-feeding over a long period, can cause a marked loss in weight and may

result in anocstrus (FAO, 1982). Severe tndcr-feeding or over-feeding may depress

the expression of heat and enhance the appearance of the condition of silent heat which

c:in be prevenied by provision of a more balanced feed enough protein, energy,

minerals mu! vitamins (Ward, el al, 1071). Mnhmoiid and Jajn. (1080) irporled low

conception rates among desert ewes grazing zinc deficient pasture in Sudan. In

another development, the time of onset of oestrus tend to be longer in oestrous

synchronized feed restricted goats than in non restricted , (Mani, ct al, 1992).

1.7 A r t i fi.ci nl _ ( I Iorni p.na I)__Con t r°.UQf_Q?stjrpus

A. Synchronization of Oestrous Using Prostnglaiulin andIjs_Syat]ieli£, Analogues:

Evidence of proslaglandin (FGF2oc) synthesis and release from the

endometrium was first revealed by Goding (1974). The natural PGF2oc, causes

normal lutcolysis through gradual degenerative changes, whereas analogues of PGF2oc

can have a very rapid and dramatic effect n progesterone synthesis in the lutein cells.

A single injection of 100 /xgm of the analogue cloprostcnol has been shown to result

in a fall in plasma progesterone concentration from 3.1 ng/ml to 0.9 ng/ml within a

period of few hours (Acritopulou. et al, 1977).

(i) Dose Levels of Prostaglandin (PGF2<x):

There has been some evidences that, oestrus response may be influenced by

PGF2« dose level. Oestrus was induced in 100% of treated females using 20 mg

PGFZoc compared with only 70% when 15 mg was employed (Hackett and Robertson,

1980). Several workers used a dose of 5 mg PGF2<x in goats (Debcnedctti, ct al,

1982); Akusu and Egbunikc, 1984; Pandey, et al, 1985). Percent of goals that showed

- 12 -

oestrus were 88%, 80 - 100%, 70 - 100% respectively. Similar results were obtained

when a 3 mg dose was used to induce oestrus in 83 - 100% of treated goats (Song and

Park, I9S4). A least dose of 0.265 mg w « used to induce oestrus in goats within 33;

30 - 75 hours (Costa, ct al, 1983).

Oestrus could be effectively synchronized by two doses of 250

Cloprostenol (Greyling and Vanderwesthuysen, 1979). A dose of 125 fj.gm was

considered lower and often insufficient to induce luteolysis. Variable doses of 62.5,

125, 250 ftg Cloprostenol given twice at an interval of 10 days, resulted in 87.5, 93.8,

100% of females showing oestrus (Greyling and VanNiekerk, 1986). These

discrepancies were suggested to be due to the ability of the corpus luteum to over

come small closes of Cloprostenol (Thorburn and Nichol, 1971; Simplicio and

Muchado, 1991). Another prostaglandin analogue carboprost from melhamine

employed in doses of 5, 6, 7 ^gm/Kg resulted in conception rates of 61, 38, 67%

respectively (Mahmoud, et ai, 1990).

(ii) Responsive Periods of Corpus Luteuin to Prostaglandin and Its Analogues

treatment:

The corpus luteum is only responsive to prostaglandin between day 4 and 14 of

the oestrous cycle (Chamley, et al, 1972, Douglas and Ginther, 1973). Haresign

(I97X, l()!S0) recommended I wo doses of iHUsia^laikiin given at ;i suitable interval in

order lo bring all animals ircaled into a synchronized oestrus as oestrus was scheduled

to occur 40 hours after the last dose and ovulalion takes place at 70 hours alter the last

dose (Aci iiojxjulou and liaresign, 1980).

(iii) Interval Between i'rostaglandin Doses:

Interval between proslaglandin doses may influence fertility. Ewe treated with

two doses of 125 /igm Cloprostenol given 12 days apart resulted in lower fertility rates

than those treated at 14 - 15 days interval (Fairnie, et al, 1977, 1978). An interval of

13 - 14 days between the two doses is optimal and should not be reduced; otherwise

acceptable fertility level to artificial insemination may not be achieved (Greyling and

VanNiekerk, 1986). However, 11 days interval in goats resulted in good conception

- 13 -

rates (Costa, ct al 1983; Alaisu and Egbunike, 1984; Pandey, et al. 1985). Poor

conception rates was obtained at 10 days intervals (Simplicio and Machado, 1991).

(iv) Interval From Prostaglandin Injection to Oestrus:

This interval was found to be longer after the first injection than after the

second. It was 62.4 VS; 55.3 hours (Greyling and VanNiekerk, 1986); 43 VS; 37.7

hours (Akusu and Egbunikc, 1984); 167.3± 13.6 VS; 100.3±8.7 hours (Panclcy, ct

al, 1985), 38+2 VS; 38.4 + 2.4 hours (Song and Park, 1984).

(v) Fertility Rnte Due to Treatment with ProstnglnnrJin or its Analogues:

Jennings, (1^75); nnd Roland, el nl (1078) r e t r i e d [xwr fcrlility rates in cwc.s treated

with Pros!;igl;uulin at both n:i!ur;il and A.!. This may be due to inlerfercncc of

prostaglandin with the efficiency of spermatozoa trans|X>rt in the cervix and into the

oviduct (Hawk and Conlcy, 1975). Poor conception rates may also result from

delayed time of insemination (Simplicio and Machado, (1991) or subsequent to lower

doses of prostaglandin (Mahmoud, et al 1990).

B. Synchronization of Oestrous Using Progesterone Impregnated Sponges

The Use of natural steroid (progesterone) in sponges may need further testing

(Gordon, 1975 a). It was found in extensive field trials that the 500 ing containing

sponges could be successfully employed in the induction of oestrus for early breeding,

and that progesterone impregnated sponges were as effective as other progestagens

(Fluorogestone acetate, FGA; Methyl acetoxy progesterone, MAP) in achieving

conception rate of 67.7% at 500 mg dose and 71.5% at 100 mg dose. However, this

high dose was viewed as unnecessary since optimal fertility in sheep was likely to be

associated with minimal dose of progesterone (Lamond, 1964, Gordon, 1971).

Other means of administering progestrone through vagina such as CIDR

(Controlled Internal Drug Rcleascr) arc used .is vaginal pessaries for synchronizing

oestrous in ewe (Welch, ct al, 1984) but with lower ovulation rate (Crosby et, al,

1983). However, this type of vaginal pessary ras no effect on any other reproductive

traits (Wheaton, et al, 1984; Knight, et al 1988).

- 14 -

Ishwar and Pandcy, (1992), obtained 100% synchronized oestrus with

progesterone impregnated sponges in Black Bengal goat. The authors (Ishwar and

Pandey, 1992) reported an interval range of 95 - 137 hours from the treatment to onset

of oestrus duration. However, Doijode, et al (1992) reported an interval of

80.52 + 3.04 hour and duration of oestrus period of 24.94 + 0.78 hours. Priming goats

in the tropics with progesterone intravaginal sponge give a response of 35 - 50% to

gonadotrophins treatment (Rosnina, et al, 1992).

>- Progesterone impregnated silastic intravaginal device (6.75%) was used to

synchronize oestrus in buflaloe followed by injection of 5 mg oestradiol 17 B at time

of insertion of the device (Rajamahendrait, et al 1980). The authors (Rajamahendran,

ct al 1980) reported mean interval to oestrus after removal of the device to be 102 + 10

hours from 77% of buffaloc cows showing oestrus and concluded that the 12 days

progesterone intravaginal device is effective in inducing fertile oestrus in buffaloe

cows.

C. Synchronization of Oestrus Using Progesterone/Progestagen and PregnantMare Serum Gonadotrophin (PMSG)

Pregnant Mare Serum Gonadotrophin (PMSG), is usually used in conjunction

with progesterone or progestagens intravaginal sponges before or after removal of the

sponge in anoestrous and cycling goats respectively (Corteel, 1975; Boshoff, 1980).

The idea behind the use of PMSG, was lo induce a mild superovulation, thus bringing. ; •

llic twining percentage of less piolilic breeds ol goats and sheep to an acceptable level

(Gordon, 1975 b). A dose range of 375 - 750 I.U. of PMSG was the chosen level to

synchronize oestrous as any increase beyond this will depress rather than enhance

conception rate (Botha, et al, 1975). However, a dose of 300 I.U. has been reported

to synchronized oestrus in goats (Bongso, et al, 1982).

Repeated administration of PMSG produces antibodies against PMSG (Anti-

PMSG) causing reduced ovarian stimulation after subsequent treatments (Baril, et al,

1992 b). However, Gherardi and Lindsay, (1980) observed that ewe did not show

evidence of nnti-PMSG activity after 17 consecutive doses given at intervals of 17

days.

In a cyclic ewe, PMSG given several days prior to withdrawal of intravaginal

progesterone sponge lead to increased suoerovulatory response (Roche, 1986). This

•-. does not apply to anoestrous ewes whici showed a reduced ovulatory and oestrus

response following PMSG administration several days prior to progestagen withdrawal

(Gordon. 1969).

1-8 Non Hormonal Methods of Oestrus Synchronization

Male (Buck, Ram) teasing technique is one of the inexpensive simple techniques

used to synchronize oestrus, ovulation and kidding in the tropics (Oldham, et al,

1979). The presence of a male or its related characteristics traits (sound, smell, sight)

triggers oestrus and induces ovulation, pregnancies in does thus necessitates

vasectomized males to be kept within audible and olfactory range of does (Llewelyn

and Ogaa, 1995). Sheiton, et al, (1960), reported more than 90%:of does that came

into oestrus and ovulate at the presence of the male. Only 50% of these goats did so

at smell, sound, sight of the buck and 26.3% were affected by. only male smell

(Carrera and Hernadez, 1971; Debenedetti, et al, 1982). Higher percentage of females

became pregnant when they were in frequent contact with the male (Moore and Hall,

1992), as such a buck may act as an outside factor in initiating ovarian activitiy and

in induction of oeslrous behaviour (Westhuysen, 1978; Ott, 1982).

I'rcmaiurc rcsiimplion oloesdous activity may be elicited by buck iuiroduciion

$.<. into a group of anoestrus females (Ott, 19c 1). The duration of such induced oestrus

y, lends to be shorter and more variable than those observed later in the season (Corteel,

et al, 1982), such short cycles were observed in Creole meat-goats after buck

introduction and were associated with short luteal phase (Knight, et al, 1981;

Chemineau, 1982). Introducing male only during oestrus or leaving it always with the

females, results into early embryonic deaths of 18.5% in day one after mating 34%

from day 1 - 30, 14% from day 3 0 - 6 0 , 16% from 60 - 90 days (Alexandre and

Borel, 1938).

B. Factors Involved in The Teasing Effects

(i) Introduction of The Male:

The teasing effects of a male which stimulate both ewes and does are partly due

to phermones (Knight and Lynch, 1980 a) and partly due to olfactory cues which

constitute the main sensory input from the males at teasing (Knight and Lynch, 1980).

Phermones are the unpleasant odour secreted from the subacious glands

particularly those located on the head and neck of buck or ram (Knight and Lynch,

1980 a). The ram and buck phermones were suggested to be of the same chemical

structure and have been identified as 6-trannonenal (Smith, et al, 1984) and series of

4-ethyl branched fatty acids (Sasada, et al, 1983; Sugiyama, 1983, Sugiyama, et al,

1986). The most intense goaty odour that attracts does in oestrus was Ethyl octanoic

acid. Other related compounds possessing goaty odour include 4-Methyl-Octanoic

(hircinoiacacid) which imparts a goaty flaour to mutton and goat meat (Wong, et al,

1975) and 4-Ethyl Octzenoic acid which \-. the main compound responsible for goaty

flavour of milk (Smith, et a!, 1984).

: J

Male introduction was believed to results in a sustained increase in LH through

increase in pulse frequency and release of the gonadotrophin. Ovulation as such,

occurs due to lowered sensitivity of the hypothalamus to the negative feedback action

of oestrogen (Martin, c! al, 1980).

(ii) Continuous Exposure to Males:

Females over ex|X)sed to males, became apparently less sensitive to male

stimuli, as over exposure modifies the sensitivity of nervous mechanisms mediating

oestrous behaviours (Lishman, 1975).

1.9 Artificial Insemination (A.I.) and Hand Mating

A. A^Jvarjtages^ojrj^J^^

Artificial insemination application h. goats has potential merits that favours its

further development. The advantages of A.I. in goats include reduction of cost of

breeding, avails and extends semen of bu.ics of high genetic quality, thus speeds up

j ^ the genetic change; it minimizes spre?.«i of reproductive diseases. Conception rates

p" resulting from A.I. was 50% in Palestinian goats, 70.8 and 80% in Barbari and

^ Jamnapari breeds of India respectively TDevendra and McLeory, 1982). Conception

•p rates of 66.7%, 75%; 100% were reported following A.I. in Angora goat in Turkey

J (Ozsar, et al, 1988) and was 45% in artificially inseminated local Brazilian dairy goat

f/ (Simplicio and Machado, 1991). Aryal, et al, (1992) reported conception rate of 31 %

£ , in Kiko goats. Conception rates following A.I. of goats in Switzerland was 74.7%

| (Summermattcr, 1993); it was 58.4% in oestrus induced goat VS 49.8% in natural

I. oestrus (Siunmermatter, 1990). Optimum conception rates in artificially inseminated

% goats in France using fresh semen was 70% (Leboeuf, 1993). From these figures, itit

|v seems that A.I. in goals needs to be perfected.

t B. Methods.of Semen Collection <

I, (i) Artificial Vagina (A.V.) Method:

\ Artificial vagina simulate natural vagina in providing temperature and pressure

necessary for erected penis to ejaculate. This method of semen collection has been

f recommended by several authors (Hafu:, 1980; Memon and Ott,, 1981; Hammond,

ft 1983; Chemineau, et al, 1991) to be the best method for semen collection.

r (ii) Electro-ejaculation Method:

-:' This method is useful in shy, untrained, injured bucks or rams, disabled bucks

f and rams and those lacking Libido (Hafez, 1980). Most commonly used electrot

ejaeukitut is the Ruakura Ram Probe (RRP) described by Laiug (1979).

(iii) Another old method of semen collection was the vaginal collection in which

semen is to be collected by a vaginal spoon from the anterior vagina right after

ejaculation (Sorensen, 1979). • •

C. E . U , j i j p X ^ j

The quality of the collected semen can be assessed on the basis of motility,

concentration and morphology. Motility of sperms in undiluted semen under low

power (40 x) Microscopy was described by a grade ranging from 0 - 5 according to

- 18 -

Evans and Maxwell (1987) as 0 score means no motion and score 5 means very fast

moving spermatozoa forming strong wave like appearance. Semen samples with very

good motility i.e. score 5 containing 90',o active spermatozoa or good i.e. score 4

containing 70 - 85% active spermatozoa can be used for A.I.; and samples with a

score of < 3 (45 - 65% active sperm cells) may result in decreased fertility (Evans

and Maxwell, 1987). However, semen samples with a wave motion graded > 3.0

were reported to be used in A.I. (Smith, et al, 1978).

Concentration of semen can be determined by either consistency of the semen

or by haemocytometer. Semen consistency ranges from thick creamy with mean

number of spermatozoa 5 x 107ml to clear (watery) with 0.7 x lOVml spermatozoa

(Evans and Maxwell, 1987). Fresh semen for insemination should contain at least 150

million/ml spermatozoa (Laing, 1979). A concentration of 200 million/ml was used

by Bongso and Falimah, (1982) in goal. However, 400 - 500 million seems necessary

for ewe insemination (Colas, 1978; Smith, et al, 1978).

D. Semen Diluents (Extenders)

These are meant to increase semen volume, protect spermatozoa viability and

preserve their fertilizing capacity (Salisbury, et al, 1979). Recognized types of semen

diluents arc milk diluent (whole or powdered skimmed or ultra heat treated) and

syntheiie diluents made up of buffers (Tris or cilrate) and energy sources (glucose or

fructose) and protective agents against cold shock at 5°C (Egg yolk) and a

cryoprotective agent during deep freezing (glycerolj. The following synthetic diluents

have received particular attention, these include: Lactose/egg yolk/skimmed milk,

tris/glucose/citric acid (Laing, 1979)... '••

Ritar and Salamon, (1991) suggested the addition of 1.5% egg yolk as a safer

margin for non-washed spermatozoa.

E. Semen Dilution

Pooled semen at least from three males contribution, can be extended in

powdered skimmed milk diluent (Colas, etal, 1978). In which 11.1 gm (Laing, 1979)

- 1 9 -

or 10 gm (Evans and Maxwell, 1987) of powdered, .skimmed milk and 0.2 gm of

glucose were dissolved in 100 ml distill A water, then heated to 92°C for 10 minutes

to destroy the spermicidal action of the iactenin in milk (Laing, 1979).

The diluent is then added to the semen at room temperature (25 - 30°C) and

kept under 10 - 15°C after dilution as it has no protective action of egg yolk against

cold shock if kept below 10°C.

F.

(i) Timing of One VS Two Inseminations:

Gordon nnd Crosby, (1980) achieved conception rales in excess of 70% after

inscmin:ition of cyclic owe with lof.il s\>cnn dose of 400 - 500 million/ml ;il 56 hours

after progestagen withdrawal. The (iming, when two inseminations arc carried out

after any hormonal control of oestrous cycle differs from one author to another.

Colas, et al (1973) inseminated ewe at 50 and 60 hours while Barlow, et al, (1974)

inseminated at 48 and 64 hours, timing by Langford and Hackett (1980) have been 54

and 60 hours and that employed by McClelland and.Quirke (1971) were 48 and 58

hours.

(ii) Site of Semen Deposition

Sites of semen deposition in goats and sheep may be vaginal, intracervical or

intrauterine. Vaginal insemination using fresh diluted semen is the simplest and

quickest method but requires large does of semen (Evans and Maxwell, 1987).

Intracervical deposition of semen is easier in does as the cervix can be

penetrated in 30 - 50% of does (Evans and Maxwell, 1987). In sheep, semen can only

be deposited in the first fold of the cervix (Colas, 1975 a; Gordon, 1975 a; Smith, et

a!, 1978). However, attempt to go deeper depresses rather than enhances the

conception rate (Smith, et al, 1978).

- 2 0 -

Intrauterinc insemination achieves good results with frozen thawed semen, but

involves minor surgery in sheep during embryotransfer (Boland and Gordon, 1978).

Although insemination into uterus via cervix was very difficult in ewe that doc (Gunn,

1936). Fukui and Roberts (1976) reported its possibility in both species.

(iii) Insemination Procedure

During insemination, females have to be restrained in such a way that it

involves minimum discomfort, less stress or injuries, allows quick and efficient

location of the insemination sites as stress may impair the passage of sperm through

the ewe's cervix. (Robinson, 1973). Standard methods for restraining females for

A.I. arc over the rail niiMlunl (livans nnc' Maxwell, l')K7), ami over a .siraw bale

methods (Gordon. 1083).

G. Factoj^jVJfec£mj^

Evans and Maxwell (1987), enumerated factors at A.I. known to affect fertility

such as number of spermatozoa in inseminate dose, time of insemination, method and

technique of 'insemination and inseminator experience, type of oestrus (natural or

artificially induced), female stress, climate and hygiene of the equipment.

1.10 Ilan d_Matji ngJTechnique

This technique involves taking females in oestrus to male for mating at a

prescribed periods of time. It was regarded as expensive and time consuming,

laborous practice and less effective in getting females pregnant than paddock mating

(Belschner, 1965). Hand mating can achieved conception rates equal to that achieved

in paddock mating only when males are kept away from females until 48 hours after

oestrus synchronization (Joyce, 1972).

1.11 Pregnan c vJ

I. Gestation Period:

Gestation period is the period calculated from the recorded date of fertile mating

or insemination til! the date of kidding. Gustation periods in several breeds of goats

in the tropics was reviewed by Devendra and McLeory (1982) to range from 144 - 153

- 21 -

days (146 days in average). These variations according to the above authors, were

related to environmental factors, breed of goat, genotype of kid, size of goat, sex of

kid, litter size and order of birth. Mean gestation periods were found to be 147.9±2

days in the Sudanese Nubian goat (Kudouda, 1985), 147 days in Anglo-Nubian, British

Alphine in Trinidad (Devendra and Rankine, 1971), and East African goat in Uganda

(Wilson, 1957), 149 cays in Angora goat in Egypt (Badawy, et al, 1972) and 143, 144

in Black Bengal goats in Bangladesh and India respectively (Ali, ei al, 1973).

II. Importance of Pregnancy Diagnosis:

Early testing for pregnancy is very important in dairy goats in order to prevent

delay in rebreeding, as delay in rebreeding will cause a decrease in income due to loss

of milk and kids per animal per year. It will also assists in any investigation of causes

and treatment of reproductive disorders in a breeding flock.

III. Methods of Pregnancy Diagnosis: M,

Pregnancy testing can be done by several methods but non of them is fully

satisfactory, (Richardson, 1972 b, Davies and Chaplin, 1983). A simple and reliable

inexpensive technique that provides immediate diagnosis in small; ruminants coud be

of real value in their management (Plant, 1980).

(i) 1'iilpation Techniques

(a) Abdominal Palpation (Ballotement); .'

This technique is easier in females of low body condition lhan in fat ones.

Accuracies of 80 to 95% was reported (Pratt and Hopkins, 1975) in 90 to 130 days

pregnant female and of 90.53% was reported in 61 - 130 days pregnant female (Goel,

et al, 1989 a).

(b) Bimaniwl Rectal Probe - Abdominal Palpation Technique:

This is very reliable after 70 and S»0 days of gestation (Hullet, 1972), but

manipulation of the plastic rod in the rectum of a female may cause serious trauma

(Plant and Tyrrell, 1974). Accuracy of 91 - 92% was reported (Chauhan and Waziri,

- 22 -

1991). However, reports of the practical assessment of this method for pregnancy

diagnosis in large number of females are not yet available.

(c) Cervi&il Palpation:

This is a simple Held technique for pregnancy diagnosis in does described by

Williams (1936). A firm, almost cartilaginous cervix palpated 50 days post-

insemination is an indication of failure of conception and a very soft cervix, not easily

reached at the same period, indicates pregnancy.

(ii) Pregnancy Diagnosis By Hormonal Assay

A. Hormones of Gestation

(a) Progesterone: The plasma progesterone concentration rise significantly from

3.91+0.51 ng/ml on day 40 to 5.9+0.51 ng/ml on day 60 and remain high up to 6

ng/ml until 5 days before kidding. These values in pregnancy are typical of luteal

phase in non-pregnant goat (Llewelyn, et al 1987, 1992). The rise in progesterone

level during pregnancy could be due to stimulation of progesterone synthesis by

caprine placental lactogcn (Currie, et al, 1977; 1990) or ovine placental lactogen in

sheep which begins to be secreted from day 44 onwards (Linzell and Heap, 1968).

Since placental secretion of progesterone is independent of hypotalamic -

piiuitary ,sup|x)rt, ihc female does not require Li I from anterior pituitary for placental

steroidgenesis. The level of progesterone from placental lactogen depends on the

number of foeti present i.e. it increases with the number of ibeli (Hayden, ct al,

1980); in contrast to this, Humblots et a! (1990) did not find any significant rise in

J progesterone level according to the number of foeti. The placental progesterone alone

, could not maintain pregnancy without the support of corpus luteum in an

I ovariectomized goat (Sheidrick, et al, 1980).

i(b) Prostaglandin (PGF2<x) and Oxytocin: In goats, PGF:o; produced by the

endometrium is responsible for luteolysis (Cook and Hameida 1984). In sheep,

PGF.oc is converted to luteotrophic PGE, which is an anti-!uteoiytic agent in ewe

(Ellinwood, et al, 1979).

- 7.1 -

Oxytocin, apart from the pituitary, is also secreted by the large granulosa ceils

which constitute the precursors of the corpus luteum (Kiem, et al, 1989). The

luteolytic action of oxytocin is possibly mediated via uterine PGF2cc synthesis (Cook

and Hoemida, 19S4).

(c) Estrone Sulfuie: Goat estrone sulfate level in the blood can be detected from

around day 50 of pregnancy (Chaplin and Holdsworth, 1982) and around day 70 - 80

in ewe (Tsang, 1978). High values of more than 1.1 ng/ml in the blood or urine are

strong evidence for the presence of viable foetus. High levels at more than 90 days

of gestation were positively correlated with the number of foeti and those does with

marked decrease in estrone sulfate at day 94 - 108 of gestation, delivered a stillbirth

and decomposed foeti (Rcfsal, et al, 1991).

(ti) Testosterone: Cook, (1990); observed that testosterone level,during oestrus in

the peripheral blood of goat showed high walues at the onset of luteolysis and reached

peak values between day 10 and 18 after nating. These findings could indicate that

testosterone may play a role in Juteolysis as well as an indicator of the mechanism

involved in maternal recognition of pregnancy (Cook, 1990).

(e) Oestradiol -17fi: Prandi, et al, (1986) found that, Oestradiol concentration was

always greater in the mammary venous blood (Itan in Ihe arterial blood in pregnant

goat at day 60. Biopsy from ihe gland showed conversion of oestrone to Oestradiol -

up.B. Hormonal Assay Methods for Pregnancy Diagnosis

A rapid immunoassay (both radio immunoassay RIA and enzyme immunoassay

EIA) were suggested as rapid progesterone and other hormones assay for pregnancy

diagnosis in goats on day 21 and 42 after mating with accuracy of 93.3% and 93.7%

achieved at day 21 and 42 respectively (Espechit, 1992). '

RIA and EIA can be used to assay progesterone, estrone sulfate, pregnancy

specific protein binding (PSPB), Oestrogen in milk or blood (plasma/serum) and in

hairs. Van de wiel, et al, (1986) found th; t, at day 21 after mating, the value of milk

- 24 -

progesterone were 127 and 6 n mol/1 for pregnant and non-pregnant goats respectively

and that with adoption of a minimum level of 32 n mol/1 for positive results, the

accuracy of positive pregnancy diagnosis was found to be around 80% and negative

diagnosis was 100%. A serum progesterone concentration of 1.5 ng/ml is considered

positive pregnancy (Jan. et al, 1980; Ozscar, et al, 1988), and the accuracy of

delecting pregnancy and non-pregnancy by RIA were 75% and 80% respectively

(Fleming, et al, 1990). However, accuracy of detecting non-pregnancy was reported

to be 100% (Chauhan and Waziri, 199!). :

Hair progesterone content could aiso be used for pregnancy diagnosis (Dazhi,

et al, 1986). A value of 9.85±1.20 ng/50 mg hair was found at day 120.

Serum progesterone concentration vas significantly higher in,pregnant females

carrying 2 and 3 foeti than 1 foetus 19.20, 29.9 and 9.2 ng/ml respectively as reported

by Chauhan and Waziri, (1991).

Tamanin and Chlesa (1986), reported the use of oestrone level and its

conjugates1 in scrum as s reliable method for pregnancy diagnosis in goats from day 55

of gestation, and its measurement at day 45 after mating can be used for differentiation

between pregnancy and pseudo-pregnancy.

Hoilz (1993) made use of oestrogen concentration in faeces as an indicator forpregnancy as the value of oestrogen increases markedly from the 5th week of

pregnancy. Values greater or equal to 74 ng/ml were considered as jxjsitive diagnosis.

The accuracy of pregnancy diagnosis using this criterion was 30, 90, 91 - 95% at 6,7,

and 8-12 weeks after mating, and the patten of oestrogen concentration in faeces was

similar to that in the blood serum.

Huniblot, et al, (1990) suggested the use of RIA for the diagnosis of pregnancy

or hue embryo mortality in goats, as PSPB profile in goats are similar to those found

in cuss's throughout pregnancy.

- 25 -

between pregnancy and pseudo-pregnancy.

liultz (1993) made use of oestrogen concentration in faeces as an indicator for

pregnancy as die value of oestrogen increases markedly from- die 5ill week of

pregnancy. Values greater or equal to 74 ng/ml were considered as positive diagnosis.

The accuracy of pregnancy diagnosis using this criterion was 30, 90, 91 - 95% at 6,7,

and 8 - 12 weeks after mating, and the pattern of oestrogen concentration in faeces was

similar to that in the blood serum.

llumblot, et al, (1990) suggested the use of RIA for die diagnosis of pregnancy

or late embryo mortality in goats, as P3PB profile in goats are similar to those found

in cows throughout pregnancy.

(iii) Ultrasonic Techniques for Pregnancy Diagnosis

(;j) Amplitude Depth Ultrasonic (A-Mode Technique):

This method seems to be more accurate between days 60 and 90 (Williams,

1986), 61 - 130 days (Goel, ct al, 1989) of gestation with accuracy of 91.9%. The

machine detects tissues densities and fluid filled uterus.

(b) The Doppler Ultrasound Technique:

This lediniquc deiecis foetal pulse rales after 50 - 60 days (Williams, 1986.

Bcrnardi and l-'agandcs, 1992) with accuracy of 99% lor pregnancy and 89% lor non-

pregiianey.

(c) Real-Time (B-Mode) Ultrasouid Scanning:

A 5 Mil, recta! probe of echotomography can be used between 25 to 100 days

after mating in goats as in sheep with 97% degree of accuracy (Baronet and Vaillan

Court, 1990). Twin pregnancy could be detected at day 31 of pregnancy (Ruseel,

1990), Tainturier, el. al, 1993).

- 2 6 -

(vi) Other Methods of 1'regnancy Diagnosis

(;i) Carpus I Ait cum Biometry and Weight:

Chungaih and Sharsna (1992), conclatcd the diameter and weight of the corpus

luteum with stage of pregnancy. The diameter increases with advancing pregnancy.

It was 8.17 + 0.29 mm at 15 - 30 days a ter mating and 12.5±0.05 mm at 121 - 150

days after mating.

(b) Vaginal Cytology:

Verma, et al, (1990), reported that, confirmation of absence of pregnancy and

exhibition of oestrus in goats could be reached by staining vaginal :,;..ear with

papanicolaous technique that revealed more clumps, superficial, intermediate and

parabasal cells with sharp round or rolled border with acidophilic cytoplasm.

(c) Absence of Return to Oestrus:

Hoidsworth and Davies (1979), hold on the non-return to oestrus 22 to 26 days

after insemination or mating in goats as a reliable sign of pregnancy.

CHAPTER TWO

MATERIALS AND METHODS

- 27 -

2 • 0 MAJM1UAJS, A N i l MJSIHQDS

2.1 £tiKJx_Area

The experiment wa.s conducted at Hclat Kuku in Eastern Nile Province at the

main centre for goats upgrading project from July to December 1995. The centre was

surrounded from the south by the Fattening Research Centre and from the east by the

Federal Artificial insemination centre, a land and a canal running from the Blue Nile

southwards to the northeast bordered.

2.2 Eiiv] r ojjjm £0J*)jCondil.tl.Q.QS«

Throughout the experimental period (July - December), the environmental

conditions were as shown in Table 1.

2.3 .fcAiieri m enjta I. A nj jrmijs:

A total of 150 females consisted of local indigenous goats and crosses of mixed

blood between locan and exotic pure breeds were purchased from Helat Kuku market

with an objective of cross breeding them with Saanen bucks to produce generations

with 75% Saanen blood. They were of various categories ranging from lVi - 5 years.

From this flock, 34 females Nubian goats were selected for experimental.

Table 1: Environmental Conditions During Experimental Period (July - December, 1995)

1

July

Ang

Sent

Get

{ Nov

Dec

Mean Temp. "C

Max.

37.8

37.8

39.8

38.6

36.2

32.1

Min.

26.2

25.9

?S.6

36.5

28.1

15.9

Mean Total

Rainfall (mm)

49.9

11.3

9.7

5.1

0.0

0.0

Mean Relative Humidity %

06 u.m.

60

58

50

51

50

48

09 a.m.

44

43

35

37

32

31

12 p.m.

39; 35

28

30

28

25

00 p.m.

50

51

50

49

51

49

from Shambat Mdcorological Station 1995"

- 2 8 -

2.4 Husbandry:

2.4.1 Aiding: The ages of the selected females were estiinated by dentition according

to McNitt (1983) which ranged from Wi to 5 years. Weights ranged from 10 to 28

Kgs. Parity was determined according to the external features, shape and structure of

udder and teats. Freedom from foreign body and pregnancy was done by abdominal

palpation, positive ones were replaced by others from the main flock.

2.4.2 Housing: The animals were housed in an open side shed. The roof was 3.0

meters in height and was constructed with corrugated and galvanized zinc sheets. The

floor was 1.0 meter above the ground, made of concrete (Plate No. 2).

- 29 -

Plate No. 1: Physical Characteristics of Sudanese Nubian Goat.

IF

liiiiiiiiiiiiiiiii::?:::r::x::::¥::::::>:

illl.

Plate No. 2: Pens for Intensive Housing of Nubian Goats.

The selected females were put into three separate pens each measures 8.5 x 5nr

with 1.25 m1 floor area per goat which was not consistent with die floor space

recommended for goats (2.16 rrr) by Devendra and McLeory (1982), or 1.5 n r by

Grunsell and HiJI (1980). Each pen was provided with water troughs and feed troughs

of 3.0 x 1.0 nv for 10 animals (0.3 nr/goat). The house was well ventilated, but not

well protected from rain when wind blew in from north-east direction.

2.4.3 Health: The goats were treated against ecto and endo-paraskes using Ivomec

super (MSD.AGVlZT.UvSA) 1.0 ml/50 Kg, injected intramuscularly; and were also

given Coccidostat supcrmed TS (Bremer-pharnia. GMBH.W.Germany) in drinking

water 1.0 gm/25 - 30 Kg body weight for three days. Tyloject 20% (Brerner-

pharma.GMBH.W.Germany) intramuscular cover was given in a dose of 1.0 - 3.0

ml/animal/day for five days. There was neither previous history of vaccination against

the maior infection.1; diseases, nor did screening for common abortive, diseases, was

done.

2.4.4 Feeding: The ingredients and composition of the rations offered throughout me

experimentation period were shown in table (No.2). Ration (1) was offered before the

adaptation period, then replaced by ration (2) during the adaptation period and for the

remaining part of the experiment. The concentrates were fed-, at the rate of 0.5

Kj',/lic;i(!AI;iy acconlinj: in rceoumit'iuhi'Dii for IVcdiuj-1, com;aurau\s to yearling and

pm'Minii dry dues (DcvaiwQ and McLeory, 1982) which ran^e from 0.2 - 0.7

Kg/hcad/day. The animals were allowed free access to salt lick. Sorghum stover

(Abu 70) or berseem (medicago sativiO were given once every other day. Water

troughs were fa led with fresh water twice daily in the morning; and afternoon. The

animals were allowed for exercise and to graze twice a week .in an adjacent field

previously cultivated with berseem and surrounded by Mesqoite shrubs (Prosopis

chilensis).

2.5 Experimentation

2.5.1 Adaptation Period:

The animals were divided according to age categories into three groups so that

each group was composed ol" representative from each age category (1 Vi - 2, 2 - 3, 4 -

5 years) and were then adapted for three weeks during which they were monitored for

oestrus, Oestrus signs and duration. ..

Table Ingredinets and Composisvon of Ration 1 and 2

Ration

Ration 1

[L. . _ ___^mlT,

i Ration 2

Ingredients

Wheat Bran

Molasses

Groundnut Cake

Urea

Salt

Soybean

Yellow Corn

Wheat Bran

Mineral Mixture

Salt

Ration 1: Manufactured by Kuku Fatteuing Research Centre.

Ration 2: Manufactured by University ot' Khartoum Feeds Factory and Poultry Farm.

Both rations (1 and 2) were analyzed in central animal nutrition research laboratory

Kuku research centre.

- 32 -

2.5.2 Treatment:

Aii the liiihiMi'.o proupfi wurc randomly assigned to 4.0 different treatments as follows:

1) Treatment (A): This is conduced in control group untreated and observed for

natural oestrus' and hand mating.

2) Treatment (H): Each female goat in t.hc group was injected with 125 /<m (0.5

ml) cstrumatc injection (Cioprostcnol), Copper LTD; Lot 053080 B intramuscularly.

The second dose was given 13 days from the first one. Number of females showing

oestrus following the first and second injections and interval from injection to

appearance of oestrus, ocslnis signs and duration of oestrus period were recorded.

3) Treatment (C): This treatment involved introduction of progesterone

impregnated sponges into the vagina. The sponges were impregnated with 40 fng of

progesterone (Aivefra.GMBH.Batch No. 24200) covering all the sponge matrix by

using a disposable syringe. Impregnation of the sponge could also be done after

insertion of the sponge in the bevelled cid of the applicator tube to prevent squeezing

out of flic progesterone. Equipments were as shown in Plate No.3.

The sponges were inserted intravr ginaily according to procedures described by

Evans and Maxwell. (1987) as follows:

1. The vulva and posterior vagina of the female were washed by acriflavin solution

before treatment.

2. The applicator (Plunger and bevelled tube) was also washed and disinfected by

acriflavin solution.

3. The impregnated sponge was then inserted into the bevelled end of the tube

with its anterior part dusted with antibiotic powder (Strcptopan), then the tube

was lubricated with vaseline before insertion.

4. The animal was lifted from the hina quarters to stand with the fore limbs, head

and neck between the assistant's lt^-s (Plate No. 4). The animal was held calm

and immobile. Then the tube was inserted gently, without much force and with

gentle twisting at the entrance of the genital tract.

5. After securing the sjx>nge in the anterior vagina, the tube was withdrawn

backwards 2 - 3 cm and the sponge was then pushed with the plunger. The

applicator was then removed leaving a 1 5 - 2 0 cm string protruding from the

vulva (Plate No.6).

6. The sponge was left for 16 days, then removed by pulling the string

downwards.

4. Treahnent (D): Involved insertion of progesterone impregnated sponges

huravaginaliy as in treatment (C) plus intramuscular injection of. 300 I.U. PMSG

(Folligonan Inter Vet) two days before rsmoval of the sponge, i.e. day 14th. The

number of females retaining the sponges until removal, females showing oestrus,

interval from the sponge removal to oeslrus, oestrus signs and duration of oestrus

periods were recorded for both treatment* C and D.

5. InirodudioM «>j" Malr.s:

A vaseclomi/.cd Nubian buck and a-spcrmic sexually active Saauen buck were

introduced lo the treated groups following termination o( each treatment lor detecting

females in oeslrus as well as initiation of oestrus.

- 34 -

INo. 3: liquipmcnl lor Application of Vaginal Sponge and A.I.

^^^^W^V^^SMlMii': § KpSX^Mi v:

lute No. 4: Restraining Doe lor Sponge Application and A.I.

1 s

Plate No. 6: Iniravaginal Application of Uie Impregnated Sponge.

- 3 7 -

2.5.3 Artificial Insemination and Hand Mating

A) Semen Collection:

A clean, dried, sterilized standard artificial vagina for small ruminants (IMVQ

40) was used for semen collection (Plate No. 3). The jacket was half filled with water

at 48 - 50°C through a tap fitted at the side after pulling the inner ruber liner with the

left hand to give a space. Pressure was adjusted by inflating air through an open

valve. The open end of the A.V. was lubricated with Vaseline into a depth of 3 cm.

Optimum temperature of A.V. before collection was 42 - 45°C and frequently checked

by a sterile thermometer. The collecting tube (MANUPYR) fitted at the other end was

maintained at temeperature of 30 - 37°C to avoid coldshock to the collected

spermatozoa.

A teaser or female in oestrus was introduced to the bucks and the collector took

crouching position at the right side of the teaser and held the A.V. along its flank with

an open end facing the male at an angle of 45°C. The penis of the buck upon

mounting was directed towards the open end of the A.V. Ejaculation was indicated by

a strong upwards and forward's thrust. ,,;

The valve and tap were then opened to release the pressure inside the A.V. and

replacement of water. The tubes containing the semen were placed in a water bath at

30°C. A jxx)l of Iwo cjactil.iles from each buck (three bucks) was prewired according

to Colas, et al, (1978) after evaluation.

B) Semen Evaluation

(i) Colour and Volume:

Semen volume and colour were read directly from the graduated tube and

recorded in millilitres. The consistency was described as creamy, milky, opaque and

watery depending on the content of spermatozoa and seminal plasma.

(ii) Mass Activity:

A drop of semen from each buck was placed on a clean warm *' \ . glass slide

and examined without a cover slip under 40 x of the olympus BH-2 Microscope.

Scoring system of mass activity was as tabulated by Evans and Maxwell, (1987),

ranging from 5 - 0, (Table No.3).

C) Concentration of Spermatozoa

A Burker-design Chamber haemocytometer was used to count spermatozoa.

Semen samples from the pool was diluted to die rate of 1 : 625 as follows: 0.1 ml of

semen was added to 0.9 ml of physiological saline solution, 1 : 10 to make dilution

(A) 0.8 ml from (A) was added to 49.2 ml of the saline to give 1 : 625 dilution (B).

A small drop of die diluted semen was deposited at the edge of the groove of the

haemocytometer under a coverslip and-feft for sometimes to allow spermatozoa to

settle down on the grid. Total number of spermatozoa was counted from five large

squares (each square has 16 small ones). Concentration of spermatozoa/mm3 was

calculated by the equation: l

No. of spermatozoa/mm3 = N x 4000 x D/80

N x 50 x D

N x 50 x 625

N x 31250

No. of sperm cells/ml = N x 31250 x 103

Where N was the number of sperm cells per 80 smaller squares. 4000 denotes the

reciprocal of the volume of the smaller squares. D stands for the rate of dilution.

- 3 9 -

Table 3: Scoring System for Mass Activity (Wave Motion)

Score

5

4

3

2

1

0

Class

Very Good

Good

Fair

Poor

Very Poor

Dead

Description

Dense, very rapidly moving waves, containing 90% or'activespermatozoa.

Vigorous movement with about 70 - 85% active-spermatozoa.

Slow moving >»'aves individual active sperm cells observed around45 - 65%. ;.,

No wave, poor movement of sperm cells with 20 - 40% of aliveactive sperm cslls.

Weakly moving with few living (10%) spermatozoa.

All spermatozoa are dead, motionless.

D) Semen Dilution '-'•

Skimmed powdered milk was the diluent used for fresh semen of bucks for

short term storage. It was prepared a day before the collection of semen. 10 gms of

the powder and 0.2 gms of glucose were dissolved in 100 ml distilled water stirred by

a glass rod and heated to 92 - 95°C in a waterbath for 10 minutes and incubated for

24 hours to adjust the PH to that of semen (7.0). The dilution was done with both the

diluent and semen in a water bath at 37°C.

Percentage of active s|)cniialozoa were assessed again alter dilution. The

diluted semen was then filled into a 0 5 ml French straws sucked by mouth after

homogenisation. The open side of the straws were sealed with a poly Vinyl alcoholic

powder.

(iii) Insemination Technique: The treated does whether observed in oestrus or not

after treatment termination were inseminated first at 24 hours and .then after 48 hours

(double insemination). Site of semen deposition was intra cervical or uterine as long

as the cervix could allow the passage of the straw. A volume of 0.5 ml fresh semen

containing 200 x 106 active spermatozoa were used.

- 40 -

Doe to be inseminated was restrained with the hind quarters held up and fore limbs on

the ground between die assistant's legs as described by Gordon, (1982) in sheep (Plate

No. 4). A lubricated plastic speculum \vi_h a light-stick was introduced into the vagina

to visualize the cervix, then the semen was deposited by pressing the inseminatiojn gun

(Plate No. 5).

2.5.4 Return to Service

Inseminated does were monitored for return to oestrus after 21 - 35, 60 - 90

days by the aid of vasectomized buck introduced directly after insemination. Those

does returning to oestrus from group B, C, D were hand mated as described by

Belsclmer (1965); Robinson, (1974); Jenning, (1976, 1977).

2.5.5 Conception Rates ,,

Conception rate based on non-return rates following first insemination (doubled)

were determined after 30 - 60 days post, insemination.

-41 -

Plate No. 5: Insemination Procedure

- 4 2 -

2.5.6 Pregnancy Diagnosis (PD)

I. Early Pregnancy Diagnosis by Progesterone Radio Imifiuno Assay (RIA)

(i) Blood Samples:

Blood for serum were taken from jugular vein using vacutainer tubes with no

anti-coagulants. Harvested samples stored at 4°C (in an ice batli) immediately after

collection and then centrifuged at 3000 'I for 15 minutes. Harvested sera samples

were stored at -20°C until assayed. ;

(ii) Progesterone RIA Kit:

The kit was assembled by the Animal Production Unit, FAO/IAEA, Agriculture

and Biotechnology laboratory, Seibersdorf, Austria (1996). It supplied reagents

enough for 500 samples which include:

1. Progesterone antibody-coated tubes; fi

2. Buffered 125 1 - progesterone;

3. Progesterone standards in blood/plasma in nmol/1 (0.0, 0.3, 1.6, 6.4, 15.9,

31.8,63.6) ' ,,>

4. External quality control samples (EQC). .,

(iii) Radio Immuno Assay Procedure:

The procedure was done according to FAO/IAEA assay protocol version 3.1 (1996),

1. Samples and oLlicr components were brought at rootii-icmpcruUirc beforestarting the assay.

2. Antibody coated tubes for samples, standards IQC and EQC were labelled.

3. The sera samples, standards and quality control were mixed with a vortex

mixer.

- 4? -

Percent BoundCo o

>

\

\

•V

ri

f t -

6o

CM

Fig No. 8. Standard Curve.\

a

•ou

- 44 -

4. The standards, quality control and samples were pipetted (100 ul of each) into

the bottom of corresponding tubes using a microlitre pipette fixed with

disposable plastic tips.

5. 1.0 ml of 125 1 - progesterone was pipetted to each tube within 5 minutes of

adding the standards, quality control and samples using multi-dispenser

instrument.

6. The tubes were then covered with aluminium foil and'incubated at .room

temperature for 3 - 4 hours.

7. All tubes in a foam decantation rack except total count, were vigorously

decanted into an appropriate radioacrve waste disposal pool pr container. Then

the rack was held upside down, tubes were stroke sharply downwards on

absorbent paper and allowed to drain for 2 - 5 minutes to remove residual

droplets.

• v'

8. The radioactivity in all the tubes were counted in a gamma .counter for a fixed

time (one minute).

9. Maximum percentage binding in the assay (R Max) was calculated using

formula.

B Max - Average CPM of Zero Standards (13,) x 100Average CPM of TC ,r

10. Percent binding values for all standards, samples and quality control tubes were

calculated using equation: <i

B/Bo = Average CPM of Standards/Samples/OCAverage CPM of Bo

11. Percent bound (B/Bo) and progesterone standard concentration were ploted on

vertical (Y) axis and horizonal (X) of Logk-log graph paper and the

progesterone concentrations of the samples and quality control were measured

by reading their percent bound values and interpolating from the standard curve

to the progesterone on the X-axis (Fig. 8).

II. PD By Abdominal Palpation Technique

Pregnancy was diagnosed at day 90—110 post-insemination by abdominal

palpation technique described by Hopkins, (1975); Plant (1980) confirmed then by

maternal physical and physiological changes on day 120 (Laing, 1979).

2.6 Statistical Analysis

Analysis of Variance (ANOVA) used in randomized complete block design was

used. Duncan's multiple range test and student "t" test were used to test significance

between means.

CHAPTER THREE

RESULTS

-46"-

3.1 Oestrus Parameters

3.1.1 Response of Does to Different Treatments

The response of 30 does to different hormonal treatments are shown in table 5.

Four does (control or treatment A) showed spontaneous oestrus during the adaptation

period. The response was 100%.

Following the administration of cloprostenol (treatment B) first dose; 6 (60%)

out of 10 does responded, one of the 10 does aborted a two months old foetus. Does

which failed to respond plus those responded to first dose (all 9 = 100%) showed

positive response following administration of the second dose of cloprostenol

(Treatment B) injected at 13 days apart.

Oestrus was observed in 7 (70%> of does out of 10 does following removal of

progesterone impregnated sponges (Treatment C) on day 16. Similarly, 7 (77.7%)

does out of 10 showed oestrus following termination of progesterone impregnated

sponges + PMSG treatment (Treatment D). There was a significant difference

between treatment B VS C and D (P<0.05). Treatment B was similar to A in the

induced oestrus.

• i

3.1.2 Insertion, Retention and Removal of Progesterone Impregnated Intra

Vagiiuil Sponges: Table 4 describes easiness of insertion and removal of intra vaginal

impregnantcd sponges and their retention within 16 days following treatment C and D.

Nineteen (95%) out of twenty does assigned to treatment C and D, retained the

sponges insilu until removed on day 16 of insertion. One doe aborted on day 12 and

consequently lost the sponge. Removal of the sponges were easier in pluriparous does

and became more difficult in primiparous and nulliparous does; Milky fluid was

observed following removal of the sponges. :<;i

3.1.3 Interval from the end of treatment to onset of Oestrus: >

The mean interval from termination of treatment to first signs of oestrus was

not statistically different (P>0.05). Tli<; mean and standard error (Mean±S.E) was

§

- 4 7 -

52.6±11.1 hours in treatment B (Table 5) and 53.3±17.6 hours in treatment C,

52.3 ±14.3 hours in treaunent D.

Table 4: Number of Does Retaining the Sponges and The easiness of Insertionand Removal of the Sponge.

Treatment

Progesterone Impregnated

Intra Vaginal Sponge

(Treatment C)

Progesterone Impregnated

Intra-vaginal Spongeplus PMSG

(Treatment D)

Parity

Nulliparous

Primiparous

Pluriparous

NuJliparous

Primiparous

Pluriparous

Insertion andRemoval

Difficult

Easy

Easier

Difficult

Easy

Easier

Number Retainingthe Sponges upto16 days

4

3

1

4

3

3

3.1.4 Duration of Oestrus Period '

There were significant differences between treatments (P<0.05) as far as the

duration of oestrus period was concerned (table 5). The mean ± S.E. of oestrus

period was 36.6±3.2 in treatment A, 52.6±4.8 hours in treatment B and 38.6±7.1

hours in treatment C and 52±7.8 hours in treatment D.

3.1.5 Frequencies of Manifestation of Oestrus Signs

The frequencies of manifestation of all oestrus traits (Table 6) peaked during

the time of oestrus (time, 0 hours) in all treatments with exception of bleating, and

mounting (Fig. 7, a,b,c,d). Does countinued to display tail wagging after oestrus

(+24 hours) with some little insignificant variation between treatments (Fig.7. e). The

does showed signs of interest in the male and frequently urinated in front of the male

before oestrus (-24 hours).

- 4 8 -

Table 5: Does Responded to Treatments x Mean interval from Treatment to Onset ofOestrus (hours) and Mean D" ration of Oestrus Period (hours).

Parameter

Does responding totreatments

Number (%)responded

Mean intervalfrom Treatmentend 10 Onset ofoestrus (hours).

X ± S.E"

Mean Duration ofoestrus period(hours).

X + S.E.

Age Group(Year)

1 Vi - 22 - 34 - 5

1 xh - 22 - 34 - 5

1 Vi - 22 - 34 -5

A

121

a4 (100%)

*

N.MN.MN.M

204248

b36.613.2

Treatments

B

333

ta1'

9 (100%)

544856

52.6±11.1

544856

a52.6±4.8

C

232'

c7 (70%)

366460

53.3,±17.5

362060

b I38.6±7.1

D

322

7 (77.7%)

6448*44

52.3±14.3

644844

a52±7.8

Not measured (N.M) ** Mean ± Standard Error (X±S.E).a,b,c, values in raws denoted by different superscript differ significantly (P <0.05).

Fig. 7: Frequencies of Manifestation of Behavioural and Vaginal Signs ofOestrus in Relation to onset of Oestrus in the Sudanese Nubian Goats.

- 4 9 -

Relative Frequency (R.F) of Incidence of Doe Behaviour inRelation to Onset of Oestrus (Time 0) in hours.

120

0 +24

D Control A iCIoprostenolB

I f Progesterone Sponges C (Progesterone +PMS6 D

Fig.7a:

Hours from onset of Oestrus

Tail Wagging during and after Oestrus. T?

- 5 0 -

Relative Frequency (R.F) of Incidence of Doe Behaviour inRelation To Onset of Oestrus (Time 0) in hours.

120

0

U Control A • Cfoprostenol B

I Progesterone Sponges C 1 Progesterone +PMSG D

Hours from onset of Oestrus

Fig.7b: Female interest in seeking on-, male and frequent urination.

-51 -

Relative Frequency (R.F) of Incidence of Doe Behaviour inRelation to Onset of Oestrus (Time 0) in hours.

o-24

D Control A iCioprostenolB

I Progesterone Sponges C I Progesterone +PMSG D

Hours from onset of Oestrus

Fig.7c: Male interest in female as sign of Oestrus.

- 5 2 -

Relative Frequency (R.F) ol Incidence of Doe Behaviour inRelation to Onset of Oestrus (Time 0) in hours.

120

-24 +24

D Control A 1 Cloprostenol B

i Progesterone Sponges C 1 Progesterone +PMSG D

Hours from onset of Oestrus

Fig.7d: Female restlessness as sign of Oestrus.

- 5 3 -

Relative Frequency (R.F) of Incidence of Doe Behaviour inRelation to Onset of Oestrus (Time 0) in hours.

o +24

J Control A iCIoprostenolB

I Progesterone Sponges C i Progesterone +PMSG D

Hours from onset of Oestrus

Fig.7e: Mounting behaviour as oign of Oestrus.

- 5 4 -

Relative Frequency (R.F) of Incidence of Doe Behaviour inRelation to Onset of Oestrus (Time 0) in hours.

120

100

80

S 603

I 40

20

0-24

r

0 +24

I Control A 1 Cloprostenol B

I Progesterone Sponges C 1 Progesterone +PMSG D

Hours from onset of Oestrus

Fig.7f: Bleating before Oestrus at night.

Relative Frequency (R.F) of Incidence of Doe Behaviour inRelation to Onset of Oestrus (Time 0) in hours.

120

-24

I Control A idoprostenolB

i Progesterone Sponges C I Progesterone +PMSG D

Hours from onset of Oestrus

Fig.7g: Characters of vulval discharges before, during and after Oestrus in Does.

- 5 6 -

Relative Frequency (R.F) of Incidence of Doe Behaviour inRelation to Onset of Oestrus (Time 0) in hours.

120

I Control A iCIoprostenol B

I Progesterone Sponges C 1 Progesterone +PMSG D

Hours from onset of Oestrus

Fig.7h: Vulval swelling and redness during Oestrus.

-57 -

Table 6: Relative Frequency (R.F) of Incidence of Doe Behaviour in Relationto Onset of Oestrus (Time 0) in hours.

Oestrus Traits

Tail Wagging.

Female interest in seekingMale and frequenturination infront of Male.

Male interest in theFemale.

Restlessness.

Mounting Behaviour.

Bleating.

Vulva Discharge.

Vulva Swelling andRedness.

RelativeFrequency fromtreatment endto Onset ofOestrus

-240

+ 24

• - 2 4

0+ 24

-240

+ 24

-240

+ 24

-240

+ 24

-240

+ 24

-240

+ 24

0+ 24

Treatments

A

10050

5010075

507525

7510050

255025

1002525

5010050

10050

B

10045

5510077

5550 *33

557745

2323~23

77 '>23- j23

23 i100 ;45M

100 |23 ;

i

C

10085

5710028

574228

577229

142914

852914

5710029

10042

D

10028

4210030

42. .4228

4010028

142814

1002814

2810042

10028

R.F. = No. of Doe Showing the Oestrus TraitNo. Responded to Each Treatment.

100

- 58 -

Males, on the other hand showed interest in a doe before oestrus (- 24 hours). This

interest increased during oestrus.

Bleating was intensely expressed by does before oestrus at night (- 24 hours) in the

absence or presence of the male in all treatments.

Vulva discharge was observed to begin before oestrus with low frequency (23 - 57%),

then peaked during oestrus (100%), and again declined after oestrus (29 - 50%) in all

treatments (Fig.7,g). The discharged mucus was a clear, thin, glistening fluid before

and during oestrus, then became claudy, creamy and thicker. The frequency of vulval

swelling and redness increased at oestrus and became less after 3 - 5 days from start

of oestrus (Fig.7.h).

Does were observed to be restless before oestrus day with highest frequency in

treatment A (75%) and was highest in treatment A and D and lowest in treatment C

during oestrus (Table 6).

Mounting activity was seen at lower frequencies throughout the oestrus period (Table

6, Fig.7,c). It was more clearly expressed by pluriparous and wqll bodied does than

others. They tend to mount primiparous rnd nulliparous does. P:

Depression of appetite however, was not observed, does continued to feed throughout

the oestrus period.

3.2 Fertility Measures

3.2.1 Return rates to oestrus upto day 30 - 60 following double insemination showed

no statistical significant differences (P>0.05) between treatments^-

3.2.2 Conception Rates:

Conception rates following double insemination showed no statistical significant

different between treatments (P>0.05), Table 7. i

-59 -

3.2.3 Pregnancy Rates:

Total pregnancy rates were significantly different between treatments (P < 0.05).

Pregnancy rates ranged from 55.5 - 77.8% (Table 7). It was observed that total

pregnancy rates were lower in treatment D compared with the high conception rate at

days 30 - 60 in the same treatment.

Early pregnancy (21 - 70 days) was determined from progesterone

concentrations by reading from standard curve which was the product of progesterone

radio-immuno assay (Fig. 8).

- 6 0 -

Tablu 7: Number of Does Inseminated, Returning to Oestrus, Conception Kates andPregnancy Rates and Docs Cxiiibiting Oestrus in Pregnancy.

Parameters

Number of Doesinseminated/handmated.

Returning to servicerates from day 30 - 60after insemination.

Total number (%).

Conception rates fromday 30 - 60 afterinsemination.

Toial number (%).

Actual pregnancy rateat 3 - 10 weeks (RIA).

Total number (%).

Pregnancy rates at day90-110(Abdominal palpation).

Total number (%).

Oestrus in pregnancy.

Total number (%).

Age Group(Years)

1 '/2 - 22 - 34 - 5

1 Vi - 22 - 34 - 5

1 'A - 22 - 34 - 5

11/2-22 - 34 - 5

1 Vi - 22- 34 - 5

L

3

a3

A

4

]

(25%)

I

(75%)

-

12

(75%)

-

2

7

a5

a7

1

Treatments

B

9

1

1

(22.2%)

223

(77.8%)

122

(55.6%)

223

(77.8%)

1

(14.3%)

C

10

21

3 (30%)

2

3

7 (70%)

22~

b4 (40%)

223

b7 (70%)

1

1 (14.3%)

I

8

c3

c5

D

9

1

(11.1%)

413

(88.9%)

•

12

(11.1%)

122

(55.5%)

-

a,b,c, values in raws denoted by different superscript differ significantly (P<0.05).

- 61 -

Mean Serum progesterone concentrations obtained in this study (Table 8) were

indicative of the reproductive status of the experimental goats post-insemination viz:

The pattern of progesterone concentrations in the serum of pregnant, non-

pregnant cyclic and acyclic goats were graphically presented in Fig. 9. In the pregnant

goats, the serum progesterone level bei;r.n at 2 ng/ml and then elevated to reach a

mean of 9.9 ng/ml as a maximum level and remained elevated above 2 ng/ml.

However, serum progesterone profiles of pregnant goats could not be disntiguished

between pseudo pregnant and prolonged luteal phase. On the other hand, progesterone

profiles of non-pregnant cycling goats began as that of pregnant, but progesterone level

fell sharply ( > 2 ng/ml) indicating return to oestrus and ovulation. The profiles of

acyclic goats had a prolonged period of baseline progesterone concentrations.

Table 8:

-62 -

Serum Progesterone Concen ration of the Sudanese Nubian Goats 10 Weelis PostInsemination.

Goat Status

Pregiuuu

Non-pregnant(cycling)

Non-pregnant(acycling)

Number (%)

12 (37.5%)

15 (46.8%)

5 (15.6%)

Mean Progesterone Concentration ng/mi

Minimum

4.9

2.8

0.1

Maximum

9.9

4.4

2.1

3.2.4 Pseudo Pregnancy

One pluriparous doe from treatment D was confirmed to be pseudo pregnant after

explusion of watery fluid on day 106 of pregnancy and subsequent expression of

maternal instincts.

3.2.5 Oestrus in Pregnancy

Two pluriparous pregnant does exhibited oestrus signs on day 65 and day 82 of

pregnancy (Table 7).

Fig. 9: Serum Progesterone Profiles of Pregnant and Non-pregnant Doses

10 Weeks Post-insemination.

-63 -

Serum Progesterone Concentrations of Sudanese Nubian GoatsPre-insemination (Week 0) and Post-insemination (3 -10 Weeks)

+ 106 $117 *2000

10

1947

106

117

2000

0.6

0.2

0.03

0.03

1 2.4

0.2

0.04

0.03

3.7

0.040.03

-.6

M

1 0 . 6004

0.2

0.3

1.40.4

0.1

8.2

50.2

0.2

2.9

0.2

0.2

0.1

0.03

1 0 . 1

0.04

0.8

3.7

1.30.11 ~

Weeks Post Insemination

Fig. 9 (A): Serum progesterone profiles of non-pregnant goats (control A). Lowervalues of progesterone concentration indicates 'non-pregnant goatsreturning to oestrus on the 4th week (No. 1947, 2000) and on the 6th and9th week (No. 106) or non-pregnant acyclic goat with basal progesteroneconcentration (No. 117).

- 6 4 -

Serum Progesterone Concentrations of Sudanese Nubian GoatsPre-insemination (Week 0) Post-insemination ( 3 - 1 0 Weeks)

105^1999

111

105

1999

0

3.2

0.03

2.7

3

8.2

0.04

5

4

10.1

0.1

8.8

5

7.5

0.1

10.7

6

5.9

0.9

8,5 •

7

13.2

0.11

11.9

8

_ 7.2

5.9

0.2

9

13.2

0.1,

13.2

10

_14.5_

0.5

11.9

Weeks Post Insemination

Fig. 9 Oil): Serum progesterone profile of cloprostenol treated goats showingpregnant goats (No. I l l , M6, 690, 122). Lower values of progesteroneconcentration indicates non-pregnant goat returning to oestrus on the 5thweek post-insemination (No. 1945) 7th week (No. 105) 8th week (No.1999, 107).

-65 -

Serum Progesterone Concentrations of Sudanese Nubian GoatsPre-insemination (Week 0) Post-insemination (3 -10 Weeks)

* 640+1945® 146

6401945

146

0

0.5

4.2

1.6

3

7.2

0.1

7.5

4

8.5

13.5

8.5

6

6.6

0.1

4.47

6"

6.3

0.1

5

7

8.2

0.2

6.6

8

8.8

0.1

6.5

9

13.2

0.03

8.2

10

8.8

0.2

7.5

Weeks Post Insemination

Fig. 9 (B2).

-66-

Serum Progesterone Concentrations of Sudanese Nubian GoatsPre-insemination (Week 0) Post-insemination ( 3 - 1 0 Weeks)

^107+122^690

107

\m|690

0

0.2

1

3

0.04

10.7

5

4

0.05

7.56.3

5

1.1

10.1

5.1

6

1.4

6.310.7

7

5

5.68.2

8

0.1

7,210.7

9

0.03

8.512.5

10

0.04

7.213.2

Weeks Post Insemination

Fig. 9 (B3).

- 6 7 -

Serum Progesterone Concentrations of Sudanese Nubian GoatsPre-insemination (Week 0) Post-insemination ( 3 - 1 0 Weeks)

* 1904+1946 & 1997

1904

1946

1997

0

50.8

8.2

3

10.7

5

0.04

4

3.55.5

10.1

5

55.4

9.1

6

3.28.2

10.1

7

7.25

6.5

8

4.55.95.4

9

5.29.15.4

10

6.36.6

10.1

Weeks Post Insemination

Fig. 9 (Cl): Serum progesterone profiles of goats treated with progesteroneimpregnated sponge showing pregnant goats (No. 1904, 1946, 104).Lower concentration indicates non-pregnant goats returning to oestrus at3rd week post-insemination (No. 1997, 1990) and 6th week (No. 1958)and on the 8th week (No. 1950, 1961) and on 7th week (1962).

- 6 8 -

Serum Progesterone Concentrations of Sudanese Nubian GoatsPre-insemination (Week 0) Post-insemination ( 3 - 1 0 Weeks)

-1950+1961®1958

1950

1961

1958

0.03

4.7

0.1

0.4

C.G0.1

21.4

4.4

7.5

6.3

1.14.1

6.3

0.17.5

5.9

0.25

0.1

4.44.5

0.04

C.30.1

0.2

5.50.1

Weeks Post Insemination

Fig. 9 (C2).

- 6 9 -

Serum Progesterone Concentrations of Sudanese Nubian GoatsPre-insemination (Week 0) Post-insemination ( 3 - 1 0 Weeks)

*1990 +1962 ® 104

1990

1962104

0

8.2

0.12.5

3

0.4

0.13.8

4

7.5

23.610.1

5

4.1

B.87.2

6

7.5

6.66.6

7

5

0.036.6

8

4.5

0.04

6.3

9

0.1

0.130.03

10

0.1

0.039.1

Weeks Post Insemination

Fig. 9 <C3)

- 7 0 -

Serum Progesterone Concentrations of Sudanese Nubian GoatsPre-insemination (Week 0) Post-insemination (3 10 Weeks)

1984+1949 $1982

1984

19491982

0

2.40.70.4

3

0.50.3

0.03

4

0.110.10.1

5

0.04

5.90.04

6

0.04

0.040.04

7

0.11

0.20.1

8

0.20.2

0.03

9

0.60.1

0.03

10

0.13

0.040.1

Weeks Post Insemination

Fig. 9 (Dl): Serum progesterone profiles of progesterone and PMSG treated goats,showing pregnant goats (No. 1989, 1998, 19"). Lower values ofprogesterone concentration indicates non-pregnant goats returning tooestrus on week 6th post-insemination (Goats No. 1949, 119) and week9 (Goat No. 1948), or non-pregnant acyclic goats (No. 1984, 1982).

-71 -

Serum Progesterone Concentrations of Sudanese Nubian GoatsPre-insemination (Week 0) Post-insemination ( 3 - 1 0 Weeks)

#1989+19-©1986*1948

1989

19-

1986

1948

0

2.56.3

0.61.4

3

6.69.4

0.04

0.03

4

23.66.6

0.03

0.03

5

13.28.5

1.12.4

6

7.53.4

0.04

8.2

7

10.18.5

0.14.7

8

5.4

6.3

0.040.6

9

8.8

8.5

0.030.04

10

5.49.4

0.031.3

Weeks Post Insemination

Fig. 9 (1)2).

- 7 2 -

Serum Progesterone Concentrations of Sudanese Nubian GoatsPre-insemination (Week 0) Post-Insemination ( 3 - 1 0 Weeks)

^ 1 9 9 8 + 1 1 9 ^ 1 9 8 8

1998

1191988

3.4

2.10.3

2.6

0.030.1

8.2

5.80.04

8

2.10.04

2.50.030.04

6.3

0.10.04

10.1

0.20.2

11.9

0.10.11

8.2

0.040.2

Weeks Post Insemination

Fig. 9 (D3).

CHAPTER FOUR

DISCUSSION

- 7 3 -

4.Response of f>oes to Different Treatments:

The three treatments, Cloprostenol injection (B), progesterone impregnated

intravaginal sponge (c) progesterone impregnanted intravaginal sponge plus PMSG (D)

were all found in this experiment to induce and synchronize oestrus in the Sudanese

Nubian goats. However, the number of does responding to treatment B were

significantly higher than those responding to both treatments C and D. The

significance of treatment B over C and D could be attributed to the rapid fall of

progesterone concentration resulting in the does coming to oestrus. Similarly,

Acritopulou, et al, (1977) found that, injection of cloprostenol resulted in a rapid fall

in plasma progesterone concentration from 3.1 ng/ml to 0.9 ng/ml within a period of

few hours. This is because cloprostenol transfered through blood affects directly the

lutein cells of corpus lutcum thus inhibits the secretory activity of corpus luteum and

enhances its regression.

The dose of cloprostcnol (125 /*g), given twice 13-days apart (two doses) in the

current study, resulted in induction of heat in the does in group B.(100% response).

This result was close to that reported in sheep (93.8%) by Greyling and VanNiekerk,

(1986); Fairnie, et al, (1977). The cloprostenol dose (125 /tig) used in this study was

lower than that recommended by Greyling and Vanderwesthuysen (1977) who

explained the effective .synchronization of oestrus as due to the use of two doses of 250

High progesterone concentration in the blood inhibits the release of

gonadotrophic releasing hormone (GnRH) from hypothalamus and LH from anterior

pituitary gland through a negative feedback mechanism. Oestrus and ovulation will

then commence at almost the same time following regression of corpus luteum on day

14. The result of synchronized oestrus (70%) obtained in this study with progesterone

impregnated sponges treatment (C) is in agreement with the finding of Gordon, (1975a)

in sheep in which progesterone impregnated sponges (400 - 500 mg) were found to be

as effective as other progestagens in inducing and synchronizing oestrus. Also, the

dose was not necessarily to be large since oestrus induction and synchronization and

- 7 4 -

©piimal fertility was likely to be associated with minimal doses of 30 mg, 100 mgprogesterone (Lamond, W64', <$or<&cm, W7\ ty. However, the synchronized oestrus

obtained in the present study is not in line with that reported by Ishwar and Pandey

(1992) in Black Bengal goats (100%). The discrepancy here appeared to be due to

effect of age and parity, breed, nutrition, management and climate (Andrade, et al,

1991).

The aim of injecting PMSG (300 I.U.) in treatment D, was to synchronize and

induce mild supervulation. It resulted into 77.7% response which was better than that

reported by Rosnina, et al, (1992) in tropical goats (50%) during rainy months and

37% during dry months when the goats were first primed by progesterone vaginal

sponges. The use of PMSG (300 I.U) in this study, resulted in a more predictable and

precise occurrence of oestrus which was consistent with the observation of Bongso, et

al, (1982).

Interval from Treatments to The Onset of Oestrus

The mean ± S.E. interval from the treatments to the onset of oestrus (Table 4)

were not significantly different among treatments. The interval from the end of

cloprostenol treatment to the onset of oestrus obtained in the present study (52 ±11.1

hours) was closed to that reported by Greyling and VanNiekerk, (1986) which was

55.3 hours, and the interval between the two injections was 14 days. However, the

interval obtained in the current study was less than the interval (100.3±8.7 hours)

reported by Pandey, ct al, (1985) with 11 days interval between injections, and was

longer than the interval rcjx>rtccl hy Akusu and Egbunikc, (1984) which was 37 - 43

hours with 11 days interval between injections. These discrepancies might be due to

the variations in the intervals between cloprostenol injections.

The interval from removal of progesterone impregnated sponge (Treatment C)

to onset of oestrus observed in this study (53.3 ±17.6 hours) was found to be the

shortest when compared to those reported by Ishwar and Pandey, (1992) in Black

Bengal goats (95 - 137 hours) and Doijode, et al, (1992) in Angora goats (80.5±3.04

hours); and Rajamahendran, et al, (1980) in buffalo cow (102±10 hours). These

THIS PAGE IS MISSING IN THE

ORIGINAL DOCUMENT

- 7 6 -

However, bleating observed in the presence of the male at night before onset of oestrus

(-24 hours) might tee inferred from \M concentration which appeared to peak during

the night in the current study with a surge lasting for about 10 hours (Llewelyn, et al,

1993). This could be attributed to absence of male sight at night and may indicate the

importance of male sight as a characteristic trait that triggers oestrus as male smell

alone is not enough (Shelton, 1960).

Mounting activity as shown by the pluriparous old females in the present study

was consistent with that observed by Mathew, (1989) who suggested that, this

behaviour was an expression of dominance by the higher ranking females.

Depression of appetite which was not seen during this study, agreed with the

previous observation of Abdel Aziz, et al, (1982) in Sudanese Nubian nannies, vivid

reason was unknown.

Vaginal discharge, vulval swelling and redness observed in this study were good

indicators for oestrus than behavioural changes, these were similarly reported by

Pretorius, (1977). The reason for vaginal changes being more pronounced than

behavioural changes was suggested to be due to sensitivity of oestradiol receptors in

the reproductive tract to rising oestrogen levels than the neural receptors which elicit

oestrus behaviours (Llewelyn, et al, 1993).

Return to

Docs returning to oestrus were not due to the effects of the treatments

employed, reasons might be due to fertilization failure or early embryonic mortalities

which might have resulted from some environmental and physical stresses inflicted on

the animal during insemination process, these environmetal stresses could result in the

release of high levels of andrcnal hormones (ACTH) especially cortisone which might

had altered uterine environment and made it inimical to early establishment of embryo

and consequently resulted in early embryonic deaths (Howarth and Hawk, 1966).

Furthermore, excitement due to rough physical handling and manipulation of the

genital tract during insemination can cause stress and the release of adrenaline which

- 7 7 -

opposes the action of oxytocin in the transport of spermatozoa up the uterine horns and

fallopian tubes henc« result In fertilization failure (Laing, 1979).

Conception Rates

Conception rates obtained in the current work in the different treatments were

not statistically different. Conception rate due to treatment C (7 = 70%) was close

to that (71.5%) reported in sheep by Gordon, (1975 a). On the other hand, conception

rate due to treatment D (8 = 88.9%) could be attributed to the superovulation induced

by low dose of PMSG and consequent fertilization, or may be due to failure to detect

does returning to oestrus from day 1 - 3 0 and day 30 - 60 post-insemination.

Conception rate due to treatment B in this study (7 =77.8%) was very close to the

control (3 = 75%). This is a good result and was consistent with those obtained by

Costa, et al, (1983); Akusu and Egbunike, (1984); Pandey et al, (1985) through

administration of cloprostenol two injection 11 days aparts, and did not agree with

poor conception rate obtained when the two injections given 10 days apart as reported

by Simplicio and Machado, (1991). Interval between doses of cloprostenol injections

and the ability of cloprostenol to reduce progesterone concentration from 3.1 ng/ml to

0.9 ng/ml within a period of few hours (Acritopulou, et al, 1977), might stand as a

reasonable justification for the fertility results obtained in the current study.

Diagnosis (I'D)

The Treatments in the present study had significant effect on the pregnancy rates

obtained. However, lower pregnancy rale in Treatment D at day 90 - 120 (5 =

55.5%) compared with the high conception rate at day 30 - 60 in the same treatment

(8 = 88.9%) might be atrributed to the heavy embryonic mortality that might have

occurred probably due to to inability of the uterus to support more than a limited

number of embryos (Robinson, 1980), or it could probably resulted from the effect of

leaving males with the females to detect does returning to heat (Alexandre and Borel,

1988).

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Weekly monitoring of the circulating progesterone levels of post-inseminated

goats using RIA was indicative of the reproductive status of the does. Doe that

exhibited progesterone concentration of ^2 ng/mf (12 — 37.5%) and maintained

these elevated levels during the prescribed period in this study (3 - 10 weeks post-

insemination) were considered pregnant. However, the obtained result was less than

the pregnancy rate diagnosed by abdominal palpation (22 = 64%) described by Pratt

and Hopkins (1975). The serum progesterone concentration for positive pregnancy in

the present study ( ^ 2 ng/ml) is in agreement with ^ 1.5 ng/ml reported by Jan, et al,

(1980) and 2 ng/ml reported by Restal, et al, (1990); Llewelyn, et al, (1995).

The pattern of serum progesterone profiles of non-pregnant goats (cyclic)

described in this study was similar to that reported by Mavrogenis (1988) in Damascus

goats. On the other hand, acyclic non-pregnant gouts maintain low baseline

progesterone profiles (<1.0 ng/ml) in this study, suggesting no ovulation and

anoestrus which is in agreement with the results of Thibier, et al, (1981).

Contrary to non-pregnant goats, there was no tendency for serum progesterone

levels to decline in pregnant goats. The progesterone profile was similar until the last

2 - 3 weeks when pregesterone levels of non-pregnant goat drastically declined

suggesting return to oestrous, while high variable levels were maintained in pregnant

goat (Fig.7), and this is in agreement with progesterone profile reported by

Mavrogenis, (1981). However, values during luleal phase and pscudoprcgnuncy could

not be separated from pregnant goals profiles by RIA during early gestation.

Abnormal Reproductive Functions

Pseudopregnancy:

The incidence of pseudopregnancy observed in this study conferred with that

reported by other authors (Mizinga and Verma, 1984; Mialot, et al 1991; Duguesnel,

et al, 1992) as a consequence of persistent corpus luteum, or prolonged luteal phase

(Zarco, et al, 1984) which resulted from disruption in uterine oxytocin receptors

during Iuteolysis (Jenner, et al, 1991) which might have interfered with PGF2«

release. Synergistic role played by LH and prolactin has also been incriminated for

- 7 9 -

persUtency of corpus luteum (Buttle, 1983). Another reason underlying the cause of

pseudopregnaney i# the hormonal treatment for synchronization of oestrus (Duguesnel,

et al, 1992).

Oestrus in Pregnancy

Females exhibiting oestrus during pregnancy (22.2%) in this study was

concurrent to the observations of other authors (Williams, et al, 1956; Younis and

Afifi, 1978).

Shutt, (1976) attributed this to excessive feeding on any oestrogenic greeu

fodders, and Gordon (1982) pointed to a probable inherited endocrine activity, or it

might be due to some hormonal changes provoked by the practice of putting

vasectomized males with females in early pregnancy to detect return to heat as done

in this study (Younis and Afifi, 1976). This was viewed to be against the evidence of

the literature in general (Gordon, 1982). Amount of oestrogen in the blood however,

was alleged to bring on heat inspite of the depressing effect of progesterone of corpus

luteum (Mirskaria, et al, 1943).

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This study confirmed thai, alt the treatments employed, induced and

synchronized oestrus in the Sudanese Nubian goats with cloprostenol treatment

showing significant response (P<0.05), and that the technique of insemination done

in this study seemed to be promising.

Interval from treatment to onset of oestrus did not differ much between

treatments; and the duration of oestrus period were significantly different between

treatments (P<0.05), but close to the mean range of goats in the tropics.

Furthermore, the characteristic reliable signs of oestrus and the frequencies of their

manifestation were not altered by the different Treatments applied.

The application of cloprostenol in two doses of 125 y.g given 13 days apart was

easier and can be used by any ordinary goat keeper. It has the merit of being easily

applied to a large flock of variable ages with minimal labour and time. Intravaginal

progesterone impregnanted sponges with or without PMSG were easier to apply in

parous females than in non-parous ones, however they involved much labour and strict

hygienic measures during application to avoid introduction of infection into the genital

system.

The actual pregnancy rale diagnosed by RIA and verified by kidding (12 =

37.5%) was less than that obtained by abdominal palpation (22 = 64%). However,

the results obtained by KIA including pscudopreguant gouts und gouts with extended

luteal phase could be more than those diagnosed by abdominal palpation method.

-81 -

Recommendationscoupled with a fixed time artificial insemination,

improved nutrition at different reproductive cycles and monitoring early pregnancy and

other reproductive parameters by progesterone RIA could all aid in adoption of

intensive system of dairy goats husbandry in the major towns. Further research on the

application of progesterone impregnation technique using different concentrations of

progesterone and monitoring the level of circulating progesterone absorped from the

sponge into the blood circulation in order to determine period when the sponge should

be remove, is required before recommedning this treatment for wide-scale use in the

field.

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