AZZA ALI ADAM ABUJE B.V.Sc. University of … · 1.2.2.4 Duration and Dosage of progesterone treatment 10 1.2.2.5 ... Radio Isotopes and Immunology, (CVRL), Souba, Dr. Asha

Association of Blood Progesterone Concentration with

Fertility and Infertility in Nubian Does in Sudan

by

AZZA ALI ADAM ABUJE B.V.Sc. University of Khartoum 1992

A thesis Submitted to the University of Khartoum in Fulfillment of the Requirements for the Degree of Master in Veterinary Science

Supervisor Dr. Sharaf Eldin Abdalla Makawi

Department of Reproduction and Obstetrics Faculty of Veterinary Medicine May– 2012

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DEDICATION

To my lovely kids;

Tagwa, Ahmed and Mohamed.

To the soul of my dear father, the pass.

To my dear mother,

brothers and sisters.

To my husband

To all whom I love, I dedicate this work

Azza



List of Content

Title Page List of contents …………………………………………… I List of tables ……………………………………………… V List of figures ……………………………………………. Vi List of appendix…………………………………………… Vii Acknowledgment …………………………………………. Viii English abstract …………………………………………... X Arabic abstract…………………………………………….. Xiii Introduction………………………………………………..

1

CHAPTER ONE: Literature Review 1.1 Detection and control of oestrus and ovulation……….. 4 1.1.1 Natural oestrus………………………………………… 4 1.1.2 Oestrus detection……………………………………… 4 1.1.3 Oestrus signs…………………………………………… 4 1.1.4 Oestrus duration and length……………………………. 4 1.1.5 Factors Influence the onset of oestrus………………… 5 1.1.6 Ovulation……………………………………………… 5 1.1.6.1 Time of Ovulation…………………………………….. 5 1.1.6.2 Ovulation rate…………………………………………. 6 1.1.6.3 Factors influence ovulation rate………………………. 6 1.1.7 Hormonal profile during oestrus………………………. 7 1.1.7.1 Progesterone…………………………………………… 7 1.1.7.2 Estradiole 17 ß�…………………………………………. 7 1.1.7.3 Lutenizing hormone (LH)…………………………….. 7 1.1.7.4 Follicular stimulating hormone (FSH)………………… 8 1.2 Oestrus synchronization………………………………. 8 1.2.1 Advantages of oestrus synchronization……………….. 8 1.2.2 Achievement of oestrus synchronization……………… 9 1.2.2.1 Oestrus synchronization using Prosraglandin F2a�

(PGF2a�) ………………………………………………. 9

1.2.2.2 Dose of PGF2a�………………………………………… 9 1.2.2.3 Oestrus synchronization using progestagens………….. 9 1.2.2.4 Duration and Dosage of progesterone treatment 10 1.2.2.5 Fertility after hormonal treatments…………………….. 11 1.3 Artificial insemination (AI )…………………………… 13 1.3.1 Advantages of AI……………………………………… 13 1.3.2 Methods of AI…………………………………………. 13 1.4 Gestation or Pregnancy……………………………….. 14



1.4.1 Pregnancy diagnosis………………………………….. 14 1.4.1.1 Return to oestrus……………………………………… 15 1.4.1.2 Abdominal palpation ………………………………….. 15 1.4.1.3 Biochemical technique……………………………….. 15 1.4.1.3.1 Pregnancy Associated Glycoprotein (PAG)………….. 15 1.4.1.3.2 Estrone sulphate………………………………………. 16 1.4.1.3.3 Progesterone (P4) …………………………………….. 16 1.4.1.3.3.1 Sites of progesterone production………………………. 18 1.4.1.3.3.2 Analysis of progesterone……………………………… 19 1.4.1.3.3.3 Advantages of progesterone pregnancy test…………… 19 1.4.1.3.3.4 Radio-immunoassay (RIA) for measuring progesterone

concentrations…………………………………………. 20

1.4.1.3.3.5 Progesterone profile in oestrus……………………….. 21 1.4.1.3.3.6 Progesterone and pregnancy………………………….. 24 1.4.1.3.3.7 Progesterone profile during pregnancy……………….. 24

1.4.1.3.3.8 Relationship between progesterone and litter size……..

27

CHAPTER TWO: Material and methods 2.1 Study area……………………………………………….. 30 2.2 Climate………………………………………………….. 30 2.3 Experimental animals…………………………………… 30 2.4 Husbandry………………………………………………. 30 2.4.1 Housing…………………………………………………. 30 2.4.2 Feeding………………………………………………….. 31 2.4.3 Age……………………………………………………… 31 2.4.4 Body condition score (BCS)……………………………. 32 2.5 Treatments………………………………………………. 32 2.5.1 Experimental procedure………………………………… 32 2.5.1.1 Treatment A…………………………………………….. 32 2.5.1.2 Treatment B…………………………………………….. 32 2.5.1.3 Treatment C……………………………………………. 32 2.5.1.4 Treatment D…………………………………………….. 33 2.5.1.5 Treatment E……………………………………………… 33 2.5.1.6 Treatment F…………………………………………….. 33 2.5.2 Sponges application…………………………………….. 33 2.5.2.1 Placing of sponges………………………………………. 34 2.5.2.2 Removal of sponges…………………………………….. 34 2.6 Artificial insemination of does………………………….. 34 2.7 Pregnancy diagnosis using RIA…………………………. 36 2.7.1 Sampling………………………………………………… 36 2.7.2 Equipment ……………………………………………… 36 2.7.3 Materials and reagent……………………………………. 37



2.7.4 Radioimmunoassay procedures…………………………. 39 2.7.5 Radioactivity measurement and result calculation ……... 39 2.8 Statistical analysis………………………………………..

40

CHAPTER THREE: Results 3.1 Reproductive performance in Nubian Does after

synchronization using progestagens………………………. 41

3.1.1 Non Return Rate (NRR) %................................................... 41 3.1.2 Pregnancy Rate (PR) %........................................................ 41 3.1.2.1 Pregnancy rate (%) according to progesterone

concentration at day 21 post insemination………………

41

3.1.2.2 Pregnancy rates (%) as that reveled by abdominal palpation 90 days post insemination……………………

41

3.1.3 Kidding Rate (KR) %........................................................... 41 3.1.4 Fecundity………………………………………………….. 44 3.1.5 Litters size………………………………………………… 44 3.1.6 Twining Rate (TR)%............................................................ 44 3.1.7 Gestation length/ day……………………………………… 44 3.2 Association of progesterone (P4) concentrations with

fertility…………………………………………………….. 44

3.2.1 Progesterone (P4) concentrations before treatment for oestrus synchronization…………………………………

44

3.2.2 Progesterone (P4) concentrations in pregnant and non-pregnant Nubian does at mid-period of synchronizations…………………………………………...

46

3.2.3 Progesterone (P4) concentrations in pregnant & non- pregnant Nubian Does at the day of insemination…………

48

3.2.4 Progesterone (P4) concentrations on day seven post-insemination in pregnant and non- pregnant Nubian Does………………………………………………………..

48

3.2.5 Progesterone (P4) concentrations on day fourteen post-insemination in pregnant and non- pregnant Nubian Does………………………………………………………..

51

3.2.6 Progesterone (P4) concentrations on day twenty one post-insemination in pregnant and non- pregnant Nubian Does………………………………………………………..

53

3.2.7 Comparisons between progesterone (P4) concentrations of the goats that not carried to term/ returned to oestrus and these which carried to term during synchronization, insemination and at first 21 days after insemination……

55

3.2.8 Progesterone profile during pregnancy, abortion and early 57



embryonic death…………………………………………..

CHAPTER FOUR: Discussion 4.1 Non Return Rate…………………………………………... 66 4.2 Pregnancy rate…………………………………………….. 66 4.3 Kidding rate%...................................................................... 67 4.4 Fecundity rate…………………………………………….. 67 4.5 Twining rate………………………………………………. 68 4.6 Litters size………………………………………………… 68 4.7 Gestation period…………………………………………… 68 4.8 Progesterone (P4) concentrations at the day of

synchronization ( day of hormonal application)………….. 69

4.9 Progesterone (P4) concentrations in pregnant & non- pregnant does at mid-period of synchronizations………….

70

4.10 Progesterone (P4) concentrations in pregnant & non- pregnant does in the day of oestrus /AI application……….

71

4.11 Progesterone (P4) concentrations on day 7,14,21 post-insemination, and during pregnancy in pregnant & non- pregnant Nubian Does……………………………………..

71

4.12 Progesterone profiles in does exposed to abortion or early embryonic death…………………………………………..

73

Conclusion and Recommendations Conclusion………………………………………………. 75 Recommendations……………………………………….. 75 References………………………………………………………….. 76



List of tables No

Table Page

1 Meteorological data during experimental period……………………………

30

2

Analysis of Feed, ingredients of the concentrate ration………………………

31

3 Chemical composition of the concentrate ration………………………………

31

4 Experimental design…………………………………………………………..

33

5 Serum progesterone standards supplied with the FAO/IQEA radioimmunoassay Kit…………………………………………………. ……

37

6 Reproductive performance in Nubian does after synchronizations using progestagens…………………………………………………………………

42

7 Effect of litter size and sex of concepts on the gestation length of Nubian does……………………………………………………………………………

43

8 Progesterone (P4) concentrations before synchronization in pregnant and non-pregnant does in all treatment groups…………………………………….

45

9 Progesterone (P4) concentrations at mid-period of synchronization in pregnant and non-pregnant does in all treatment groups………………………… ………………………………………………

47

10 Progesterone (P4) concentrations at the day of insemination in pregnant and non-pregnant Does in all treatment groups… ……………………………….

49

11 Progesterone (P4) concentrations on day seven post insemination in pregnant and non-pregnant does in all treatment groups……………………………….

50

12 Progesterone (P4) concentrations at day 14 post- insemination in pregnant and non-pregnant Does in all treatment groups………………………………………………………………………….

52

13 Progesterone (P4) concentrations at day 21 post- insemination in pregnant and non-pregnant does in all treatment groups……………………………………………………………………

54

14 Comparisons between progesterone (P4) concentrations (nMol/L) of the goats that not carried to term/ returned to oestrus and these which carried to term at a selected sampling periods …………………………………………..

56



List of figures N0 Figure Page 1 Semen collection& A.I equipments …………………………. 35 2 Gamma counter unit …………………………………………… 38 3 Equipments used in RIA ………………………………………. 38 4 Plasma progesterone concentrations (P4) during gestation

in group A………………………………………………………. 58

5 Plasma progesterone concentrations (P4) in pregnant does in group B………………………………………………….

59

6 Plasma progesterone concentrations (P4) during gestation in group c………………………………………………………..

60

7 Plasma progesterone concentrations (P4) during gestation in group D………………………………………………………..

61

8 Plasma progesterone concentrations (P4) during gestation in group E……………………………………………………….

62

9 Plasma progesterone concentrations (P4) during gestation in group F………………………………………………………..

63

10 Progesterone profile in goat no 2424 showing abortion… 64 11 Progesterone profile in goat no 1204 showing abortion…. 64 12 Progesterone profile in goat no 1806showing abortion….. 65 13 Progesterone profile in goat no 644…………………………. 65



�� List of Appendix No Appendix Pag

e

1 Progesterone (P4) concentrations at synchronization in does carried to full term and does not carried to full term in all treatment groups………………

109

2 Progesterone (P4) concentrations at mid-period of synchronization in does carried to full term and does not carried to full term in all treatment groups..

110

3 Progesterone (P4) concentrations at the day of insemination in does carried to full term and does not carried to full term in all treatment groups…………

111

4 Progesterone (P4) concentrations on day seven post insemination in does carried to full term and does not carried to full term in all treatment groups..

113

5 Progesterone (P4) concentrations at day 14 post- insemination in does carried to full term and does not carried to full term in all treatment groups…………………………………………………………………………

114

6 Progesterone (P4) concentrations at day 21 post- insemination in does carried to full term and does not carried to full term in all treatment groups……….

115



Acknowledgement

I am gratefully acknowledge my supervisor Dr. sharaf El Din

Makawi, Head Department of Surgery Gynecology and

Obstetrics, Faculty of Veterinary Medicine. University of

Khartoum for his guidance, continuous encouragement and deep

supervision of this work with an open heart and long patience.

I would like to thanks Dr. Yousif Hussein Elmansoury, Head

Manager of the General Administration for Animal Health

Research, Central Veterinary Research Laboratories (CVRL),

Souba, for his precious advices, valuable help and constant

support.

I am extremely grateful to my colleagues in the Department of

Radio Isotopes and Immunology, (CVRL), Souba, Dr. Asha

Abbas Elsadig, Dr. Elsid Ishag, Dr. Hussina Elbashir , Dr. Amna

Eltayb, Tahani Othman, Faroug Idris, Sabri Hussein, Awadia

Hamid and Badria Eltahir for their great help and assistance in

samples preparation and analysis.

I express thanks to Administrations, staff and labors in the Goat

Improving Project- Ministry of Agriculture- Kartoum state, and

National Centre for Artificial insemination- Ministry of Animal

wealth, Kuko, for providing experimental animals, equipment,

photos and other working facilities.



My thanks are also due to my large family specially my mother

for her great support and patience tally caring for my Childs, my

eldest sister Salha, for her financial supplies, my younger lovely

sister Igbal, my eldest kind brother Ahmed, my brothers Adil,

Tarig and Awwad/alla Ali Adam for their help and unlimited

encouragements.

My thanks extended also to my husband, my children's who

tolerated my absences.

I gratefully record my indebtedness to my best friends Dr.

Amna Eltib, CVRL, Dr. Siham Abdoun and Dr. Ishraga Elhaj,

Administration of Pharmacological Quality Control, Ministry of

Health, for their great help in doing Statistic.



Abstract

The objective of this study is to establish correlations between progesterone concentrations at different stages of a controlled breeding programme and fertility rate in Nubian goats.

Sixty Nubian does, were divided into 6 equal groups, according to age and body condition score. The first group A received no treatment and was naturally mated at natural heat, while the other groups B-F were treated with intra-vaginal progestagen sponges for 12 days. Group C was injected with eCG 48h before sponge removal. Group D was injected with eCG at sponge removal. Group E was injected with eCG and PGF2a� at the time of sponges removal and Group F was injected with PGF2a� 48 hours before sponges removal. All groups were inseminated with fresh-diluted semen 48h after sponge removal.

Blood samples were collected from the jugular vein on the first day of treatment for oestrus synchronization (ES), mid-period of hormonal treatment (day 6), day of insemination (day of oestrus onset), day 7, day 14 and day 21 post-insemination and at ten days intervals during the period of gestation. The blood samples were centrifuged, sera separated and kept at -20°C till assayed for progesterone hormone (P4) concentration using Radioimmunoassay technique (RIA).

The results for the non-Return (NRR) were 50%, 50%, 60%, 60%, 50% and 40% in groups A, B, C, D, E and F respectively, with no significant difference (P!0.05) among the groups. Pregnancy rates ranged between 30% and 50% without a significant difference (P!0.05). Kidding rate registered no significance difference among the groups. It reached (40%) in group D and (30%) in the rest of the groups. Group D showed the highest value (kid/doe) when fecundity rates were compared among the groups, whereas the rest of the groups recorded a range of 0.4-0.7, but with no significant difference (P!0.05). Twining rate was 100% in groups D and E, 67% in groups B, C and E and 33.3% in group F. Litter size was 2.5 in group D and ranged between 2.3 to 1.3 in the other groups without a



significant difference (P!0.05). Pregnancy period was significantly longer (P!0.05) in group B, 152.2±1.9 days, where it was 142±2.1 days in group A.

The progesterone concentrations on day one in both does that carried to full term and those returned to oestrus differed according to the phase of oestrus in each doe at the time of hormonal treatment. The total mean of the P4 concentrations on this day was 14.04±9.63 nmol/l and 12.91±5.25 nMol/l in does that carried to full term and those returned to oestrus, respectively, with no significant difference. At the mid-period of treatment, the total mean of P4 registered a higher concentrations that reached up to 26.92±12.90 nmol/l and 23.70±9.98 nMol/l in goats that carried to full term and those returned to oestrus, respectively, without a significant differences (P!0.05). Progesterone concentration then significantly (P!0.05) declined to its lowest concentrations on the day of insemination (oestrus), the total mean of P4 concentration was 1.71±0.86 nMol/l and 3.86±6.01 nMol/1 in does that carried to full term and those returned to oestrus, respectively. Seven days after insemination, the total P4 mean was found to increase significantly in goats that carried to full term to (33.81±9.97 nmol/ml) compared to those returned to oestrus (11.56±7.89 nmol/l). On day 14 post-insemination, the mean P4 concentrations significantly (P!0.05) increased with the progression of pregnancy in the goats that were carried to full term, the total mean registered was 35.77±15.91 nmol/l, where as in the goats that returned to oestrus it was 19.31±18.29 nMol/l. On day twenty one after insemination, the total mean P4 concentration in the goats that were carried to full term was 33.36±16.65 nmol/l where as in the goats that returned to oestrus the total mean level P4 decreased to 3.67±6.99 nmol/l with a significant difference (P!0.05) between the two groups.

During the first days of pregnancy the progesterone levels rised sharply compared to oestrous levels then, they were maintained at high levels with fluctuating peaks throughout pregnancy, until 2-3 days prior to parturition when they abruptly declined on the



day of parturition. The progesterone concentrations significantly (P!0.05) varied among the does through the period of pregnancy but were not affected by the treatments. However, four does in groups A, C, D and E recorded sudden decline in P4 levels between days 30-95 of pregnancy indicating embryonic mortality.

It was noticed that group D which showed the highest percentages in kidding rate, fecundity, twinning rate and litter size, also registered the highest P4 concentrations at mid-period of progesterone sponges treatment, day 7 and day 21 post-insemination. These results illustrated the importance of progesterone assay in detection of early pregnancy, early embryonic losses and in studying fertility indices in the Sudanese Nubian does.

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1

INTRODUCTION Animal resources in the Sudan are estimated to be around140 millions heads,

including cattle, sheep, goat and camel. They play a very important role in the

national economy. The goat population in Sudan is estimated as 41 million

heads distributed throughout the country (MAWF, 2009), their importance is

associated with their contributions to their supplies of milk, meat and skin as

well as manure. Goats are bred by a large sector of the people in the rural areas,

and are well of being (The poor man's cow) because they are owned by those

who can not afford to keep cows for meat and milk production.

Nubian goats are among the best dairy breeds in Africa especially in Sudan

where they comprise 47% of the total goat population. They could reach up to

85 cm in wither height: Males weigh 50-70 kg and females 40-60 kg. Kidding

interval is 228±17days, with a fair twining rate. High proportion of single births

(60%) occurs, while twin and triplet percentage are 30% and 3% respectively

(El Naim, 1979). Weight at birth is 2.5kg for males and 2.1kg for females.

Lactation length is 147days and milk yield is 1.5-2.0 kg/day (Muna et al, 2000).

Nowadays research activities on goat reproduction witness a remarkable

increase. They are focused on performance improvement through proper

management, feeding and breeding programmes.

Cross-breeding is practiced in the Sudan since 70s, it aimed at improving the

genetic potential of the low producing local breeds through crossing with the

internationally high producer foreign goat breeds like, Saaneen, Toggenburg,

and Anglonubian . This crossing is assumed to lead to an increase in goat

productivity thus increasing the welfare of the goat owners. Crossing between

local Nubian goats and Saaneen bucks is the most common practice in the

Sudan and the resultant cross- bred goats are seen almost everywhere. Their



2

improved milk productivity, tolerance to high temperature and adverse

conditions were reported.

There is a need for early pregnancy diagnosis in goats, reliable techniques for

early detection of pregnancy aid in culling or rebreeding of barren does

( Linonel, 1990 and Elmansoury, 2008), and provide available tool for

controlled breeding program. Inability to detect early pregnancy can result in

economic losses in milk supplies, kid production and longer kidding intervals

((Ishwar, 1995). Traditional methods used by goat owner for pregnancy testing

such as a change in behavior, drop in milk yield, tendency to fatten up and

change in body out line are not completely reliable, as they do not distinguish

between true and false pregnancy. Absence of heat could be a useful indicator,

but unfortunately many goats have tendency to show some signs of oestrus

during pregnancy. Therefore, a variety of pregnancy diagnosis methods in

goats have evolved, such as abdominal palpation, ultrasonography , and

hormonal assays (Kinne, 2006).

Measurement of concentrations of steroid hormones such as oestrone sulphate

and progesterone at specific times post-breeding provides a good method of

pregnancy diagnosis in small ruminants (Murray and New stead, 1988; Refstal

et al., 1991). Progesterone is used as an early pregnancy test (18-19 days after

insemination in ewes and 21-22 days in does). The over all accuracy of this test

is higher than 90%. It has been found to be of a significant clinical value in

females of most domestic species and, in fact, the most useful information

about the reproductive status of animals such as age at puberty, pregnancy,

and post partum ovarian activity (FAO/IAEA, 1999). Hormonal radio

immunoassay (RIA) has allowed the development of sensitive tests (Boscos et

al., 2003).



3

The objective of this study:

• To establish a base-line data for the normal progesterone concentrations

at the different stages of reproduction in the Nubian goats.

• To monitor the success of oestrus synchronization and A.I. in goats.

• To establish correlation between progesterone concentrations and fertility

rate at different stages of a controlled breeding program.



4

Chapter one

Literature Review

1. 1. Detection and control of oestrus and ovulation 1.1.1. Natural oestrus

Oestrus is the time during the reproductive cycle in animals when the females

displays interest in mating and in most species will stand to be mounted by both

sexes and mated by males. In most cases the females is about to or has just

ovulated, and is therefore pregnable (Blood, et al, 2007). Oestrus cycle is

divided into two phases, the follicular phase which is the period of follicular

growth and the luteat phase which is the period of corpus luteum functioning

(Noakes et al., 2001).

1.1.2. Oestrus detection

Oestrus detection is generally based on the criteria of sexual receptivity of the

female to mounting by the male , its in fact, the postural immobilization which

will allow mating and deposition of semen by the male into the female (FAO,

1990). Oestrus signs were monitored twice daily (am and pm) using a

vasecotomized buck, immobilization of the female at mounting by the male was

considered a sign of oestrus behavior ( Feritas, et al., 1996b).

1.1.3. Oestrus signs

The signs of oestrus could be manifested as: restlessness, frequent urination, tail

flagging, mounting other females, swollen of the vulva, intermittent discharges

of vaginal mucus, change of the colour of vulva from pale to bright pink

( Shalaby, et al., 2000 and Noakes et al., 2001).

1.1.4. Oestrus duration and length

Oestrus duration was defined as the time between the first and the last accepted

mount, within the same oestrus period (Dogan et al.,2005). The duration of



5

oestrus cycle of the dairy goats varies from breed to another, it was registered

as 28-35 hours in West African Dwarf goats (Akusu and Egbunike, 1990) , 12-

48 hours (Kadouda, 1985), and 36-52 hours (Jubara, 1996) in the Sudanese

Nubian does. The oestrus length is approximately 20-21 days and differ

according to the breed and environment (Hafez and Hafez, 2000), In (2001),

Zarkawi and Soukouti, reported that the average length of the oestrus cycles in

the Damascus breed was (21.2±1.5 days) which ranged between 19-26 days.

While in the Sudanese Nubian breed the oestrus length ranges between 19-24

days according to Kadouda, (1985).

1.1.5. Factors influence the onset of the oestrus

Oestrus onset was affected by many factors like nutrition which affects the

number of oocytes, attainment of puberty and sexual maturity in goats were

very much delayed in inadequately fed female goats , season mainly in

temperate breeds where there is cessation of oestrus during certain months,

presences of the male which stimulates the secretion of the reproductive

hormones and diseases which affect gonads and hence oestrus cycle

( Williamson and payner,1978; Greyling and Van Niekerk, 1990; Ahmed et al.,

1998; Romano, 1998a, 2002).

1.1.6. Ovulation

Ovulation is the process in which the mature graefian follicle ruptures and

release the mature ovum. It is spontaneous, and it occurs in does regardless of

whether or not mating takes place (Evans and Maxwell, 1987).

1.1.6.1.Time of ovulation

Ovulation in goats is generally reported as occurring a few hours after the

termination of standing oestrus (Riera,1982) or towards the end of oestrus

(Vander Westhuysen et al., 1985).Ovulation in different goat breeds , excluding

the Nubian has previously been estimated to occur 24 hours to 40 hours after



6

sexual receptivity. Nubian doe characteristically ovulates during oestrus,

possibly as a reflection of a more prolonged oestrus period in comparison to

other breeds.

1.1.6.2. Ovulation rate

In sheep and goat the number of ova released could range from one to many

depending upon the breed (Hunter et al., 2004). Ovulation rate (number of ova

liberated from the ovary during a given oestrus period) are quoted from 1-2 for

Angora (Shelton, 1960b) to 4 for Black Bengal does (Rao and

Bhatacharrya.,1980). In the Nubian breed as reported by Camp et al. (1982) ,

the number of ova produced by the female / oestrus was 3.1, and the ovulatory

activity in the Nubian doe was greater on the right compared to the left ovary

which was consistent with the findings in other breeds ( Wani , 1982 ).

1.1.6.3. Factors effecting ovulation rate

As described by Hunter et al. (2004), the ovulation rate is influenced by the

following factors:

i. The differences in the hypothalamic-pituitary-ovarian feed- back response

(mainly FSH and LH) lead to the differences in the number of ovulatory

follicles.

ii. Exogenous hormones administered or GnRH- agonist models in which

endogenous gonadotropin secretion is suppressed and exogenous LH and/or

FSH when administered at specific concentration in defined patterns, are useful

for elucidating the precise roles of specific hormones in stimulating follicular

development.

iii. Environmental factors such as nutrition which effects the quality of oocyte

and consequently embryo development and survival.



7

iv. Recently, the regulation of follicular angiogenesis has been shown , to be

important for the development of ovulatory follicles and maintenance of

adequate levels of progesterone to sustain pregnancy.

1.1.7. Hormonal profile during oestrus

1.1.7.1. Progesterone

During oestrus and during seasonal anoestrus, the plasma progesterone

concentrations is <1ng/ml. Progesterone concentrations values reported

during luteal phase (typically 4 to 8 ng/ml) are variable and depends on the

number of corpora lutea present and the assay procedure used. Progesterone

concentration drops 3 days before the next oestrus (Thorburn and Schneider.,

1972). Similar results were registered by Shalaby et al. (2000) when they

studied the reproductive performance of Damascus goats in semi-arid areas in

Egypt and Khanum et al. (2008), when they studied the progesterone profile

during oestrus in Dwarf goat.

1.1.7.2. Estradiol 17 .B

The level of this hormone in the blood rises from the base line of about 8 to

10 pg/ml at the beginning of standing heat only, to fall to the base line again

12 hours later ( Leyaocariz et al .,1995). With the initiation of cyclicity,

estradiol attains higher levels (7.7 ± 1.7 pg/ml) at oestrus phase and drops down

to the lower levels within 3 to 4 days post-oestrus (Khanum et al., 2008).

1.1.7.3. LH ( Lutenizing Hormone)

Pre-ovulatory peak of LH concentration is taken as being indicative of

ovulation. LH peaks appeared to precede ovulation by about 18 - 28h and it

relatively correlated with ovulation rate ( Ritar et al. 1980), LH surge was most

pronounced when PMSG was administered 48 h before rather than at sponge

removal.



8

1.1.7.4. FSH (Follicular Stimulating Hormone)

FSH is the main hormone controlling follicular growth in cattle, sheep and pigs

and its secretion is in turn controlled via the main secretary product of large

follicule(s), oestradiol and inhibin A. (Adams, 1999; Webb et al., 2004).

1.2. Oestrus synchorinization Oestrus synchronization or the artificial hormonal control of oestrus is a

valuable managemental tool that has been successfully employed in enhancing

reproductive efficiency in farm animals (Lu Meng et al., 2008), particularly in

dairy and beef cattle and in sheep ( Henderson et al., 1984; Carlson et al.,

1989; Hamra et al.,1989) . In goats the opportunity for control is greater during

the luteal phase, which is of longer duration and more responsive to

manipulation ( Wilddeus, 1999).

1.2.1. Advantages of oestrus synchronization

As in other species the control of goat reproduction offers different advantage at

farm levels and at the level of a population where genetic program could be

employed, these advantages were described by . (Karaemer 1989, Kusina et al.,

2000 and Abu et al., 2008 ) and were summarized as follow:

a. A large number of does could be bred over a short period. As a result,

management could be concentrated during a short period.

b. Does could be bred at the time that will be suitable for the best marketing

opportunities.

c. Synchronization would also allow producers to schedule kidding to take

advantage of feed supplies, labor, and upward price trends.

d. Implementation of breeding technologies such as artificial insemination and

preparation of donor and recipient does for use in embryo transfer.



9

1.2.2. Achievement of oestrus synchronization

Oestrus synchronization in does, as in cows and ewes, is achieved either

by reducing the length of the luteal phase of the oestrus with prostaglandin

F2a� (PGF2a�) or its analogous, such as cloprostenol ( Estrumate, Cooper ,

UK ) or extend it artificially with exogenous progesterone or more potent

progestagens (Molowuku, 1980; Devendra, 1983; Karatzas et al., 1997;

Amarantidis et al., 2004; Abu et al., 2008).

1.2.2.1. Oestrus synchronization using Prosraglandin F2a� (PGF2a�)

Prostagladin F2a� is often used in a combination with superovulation regimens

to synchronize oestrus by inducing complete regression of corpus luteum

(lutelysis) and hence affect oestrus initiation and was found to be more

convenient and economical method (Perera et al., 1987; Mahmood et al., 1991;

Krisher et al., 1994; Romano 1998b., Muna et al., 1998; Wideus, 2000; Noakes

et al., 2001; Bitaraf et al., 2007 ).

1.2.2.2. Dose of PGF2a�

Administration two doses of 125 to 250g cloprostenol 10 days a part induced

oestrus within 18-48 h in non descript indigenous goat breed (Perera et al.,

1987). Thus same procedure resulted in 84% Red Sokoto does being

synchronized ( Molowuku et al., 1980). Ozawa et al., (2005) synchronized

oestrus cycle in goat using a single intramuscular Injection of 0.5 ml (PGF2a� )

administered 8-10 days following the detection of oestrus.

1.2.2.3. Oestrus synchronization using progestagens

Exogenus progesterone is used to prolong oestrus cycle. The most commonly

used exogenous progesterones are FAG-Flurogesterone acetate and MAP-

Medroxy progesterone acetate ( Bretzlaff and Madrid,1989 ; Bretzlaff, 1997;

Motlomelo, 2002). Abu et al., (2008) synchronized oestrus in West African

Dwarf goats using intramuscular injection of progesterone.



10

The insertion of a vaginal sponges containing a progesterone analogues (FAG-

Fluroggestone acetate ) prolong the luteal phase where the high concentration of

progesterone inhibits the GnRH pulsteitile secretion by the hypothalamus thus

stopping ovulation until the following luteolysis ( Leboeuf, et al, 1997).

The use of PGF2a� application together with FAG increase oestrous response

rate (Dogan et al., 2005). The use of Gonadotropins such as eCG in conjugation

with intravaginal progestagen treatment, reardless PGF2 a� administration was

found to be efficient methods for oestrous induction and synchronization in

sheep and goats (Fonseca et al., 2005; Mehmet and Mehmet, 2006). Babiker et

al .,(2010), when they studied fertility in the Nubian goats concluded that

progesterone treatment (FAG) intravaginal sponges and their combination with

either eCG or PGF are efficient in synchronizing oestrus and seems to be more

convenient.

1.2.2.4. Duration and Dosage of progesterone treatment

The most widely used procedures for synchronization and/or the induction of

oestrus are the introduction of FAG or MAP impregnated intravaginal sponge

for 11-13 days ( Romano, 1996, 1998 a, b, 2002; Romano and Benech., 1996;

Romano and Fernandez Abella., 1997; Romano et al., 2000) and intramuscular

injection of PMSG at the sponge withdrawal ( Greyling and Van der Nest, 2000;

Babiker et al., 2010), or 11 days treatment with FAG impregnated intravaginal

sponges and an intramuscular injection of eCG and synthetic PGF2a�

analogous 48 h before or at sponge withdrawal (Baril et al., 1993; Freitas et

al.,1996a,b., 1997; Leboeuf et al., 2003).

Ovulation is effectively controlled by injection of PMSG around the time of

cessation of intravaginal treatment with sponges containing flurogestone acetate

(Ritar et al., 1984) or with controlled internal drug release CIDR containing



11

progesterone, with no difference in the results between the two treatments

(Ritar et al., 1989).

Motlomle et al., (2002) synchronized oestrus in goats using different

progesterone treatments, MAP(60mg), FAG(40mg) and CIDR. They were

administered for 16 days period and does received 300 iu PMSG i/m at the end

of treatment. These progestagen did not show significant differences in oestrus

response (97%) and duration (33.3 ± 13.4 hrs) of induced oestrus period .Time

of onset of oestrus is advanced in CIDR and also serum progesterone

concentrations in CIDR group where higher (P<0.05) between day 4 and 16 of

progesterone treatment compared to MAP and FAG groups. No significant

difference was observed with respect to pregnancy rate 40 days after AI.

1.2.2.5. Fertility after hormonal treatment

The use of progestagens for improving fertility, pregnancy and kidding rates

will continue to be a viable option in reproductive management of goats (Sahlu

and Goetsch, 2005). Fertility value estimated as the number of does conceived

per hundred does mated (Mari et al., 2000 ). Feritas et al. (1997) assessed the

number of goats ovulated and fertility (number of pregnant goats /number of

treated goats) by a semi–quantitative progesterone assay in blood samples

collected 5 or 6 days, 21 or 22 days after oestrus (day 0). They recorded

conception rates of, 83.9, 49.1 and 66.7; kidding rate 75.0, 45.5 and 58.8; and

litter size of 1.9±0.8, 1.8±0.8 and 1.9±0.8. Ahmed et al., (1998) registered that

fertility rate in Sudanese Nubian does treated with progesterone impregnated

sponges was 40% while Babiker et al., (2010), mentioned that fertility in

Nubian goats measured by the kidding rate was not significantly different

between different synchronized groups and were varied between 20%-35%.

Romano (2004) recorded that the kidding rate after AI in the CIDR, FGA and

MAP treatments were 63%, 65% and 63% respectively.



12

Fertility depends on the time of and efficacy of the treatment to control

oestrus and thereafter ovulation. Baril et al. (1993) noticed that fertility rate

decreased significantly in animals which came into oestrus later than 30h after

sponge removal (late oestrus ), than for goats that exhibited oestrus earlier

(33.3%-65%, P< 0.001). The frequency of late oestrus was not dependant on

the age of goats, but on the number of treatments that they had previously

received , suggesting that the late oestrus could be related to the appearance of

anti-eCG antibodies. The use of eCG resulted in a more predictable occurrence

of oestrous. One limitation of eCG after long-term use is the active production

of anti- eCG antibodies which decrease fertility in goats (Leboeuf et al., 1998;

Fonseca et at., 2005).

Low fertility rates were observed in does treated with half implants of

Norgostomet , compared to goats which received FAG sponges, while in goats

treated with whole implants the decrease of fertility rate was not significant.

Doses of the synchronization hormones affects fertility rate, this fact was stated

by Romano (1998b), who observed that fertility rates resulted from artificial

insemination after a dose of 125 and 62.5 mg Cloprostenol were 4/8 and 5/8,

respectively.

Fertility rate could be also affected by the type of semen used and by the

number of inseminations applied. Karatas et al., (1997) reported that the overall

kidding rate with fresh semen (65%) was significantly higher (P<0.05) than

that with frozen semen (53%), and the overall kidding rate was significantly

higher (P<0.001) in the does inseminated twice with fresh or frozen-thawed

semen (70.4% and 59.1% respectively) than in those inseminated only once

(48.9% and 44.9% respectively). A decrease in fertility following progestagin-

based synchronization in the absence of corpus luteum, and a fixed time

artificial insemination has being observed ( Lassala et al., 2004).



13

Season and housing were found to influence ovarian activities and hence

fertility, possibly due to changes in temperature and photoperiod. Lleweyn and

Ogaa (1995) observed that the proportions of normal cycles occurring in winter

was 87% and 77% for goats in single and groups pens respectively, falling to

62% and 37.5% respectively in spring.

1.3. Artificial insemination Artificial insemination even when used in small scale, allows links between

flocks which increases the efficiency of indexation of sire. Early and accurate

estimation of the genetic value of young bucks is feasible and semen from

identified improved males can be spread rapidly in a large number of flocks.

1.3.1. Advantages of A.I

The use of AI with spermatozoa of selected bucks has led to increase milk

production in goats. Further genetic improvement depends on the widespread

use of AI which can only be possible if the treatment is used to induce

synchronized oestrous that will result in a good fertility rate (Baril et al.,1993).

1.3.2. Methods of AI

Artificial insemination in goat can be applied by many methods such as vaginal

insemination (Paulnez et al., 2005), intracervical insemination (Leboeuf et al.,

2003), intrauterine insemination (Ehling et al., 2003), and laparoscopic

insemination (Sohnery and Haltz, 2005). Cervical insemination involves

deposition of sperms in the uterine side of the cervix, conception rate using this

method ranges from 50 to 70% depending on the season of insemination. The

conception rate is low during spring and summer due to lower sperm motility

than in autumn and winter however, photoperiodic treatment of bucks

(Delgadillo et al.,1995), which enables sprem collection all year round, may

alleviate such seasonal variations in sperm quality. The Laparoscopic

insemination methods involves the use of a laparoscope and manipulating probe



14

to aid in depositing fresh or frozen –thawed semen directly into the uterine

horns. Laparoscopic insemination procedures are described for sheep and goats

by Ritar and Ball (1991). More than 80% success rate of conception is realized

(Amoah and Gelaye, 1992).

Insemination is performed too early to enable the fertilization of ova, this

explanation is supported by the fact that a delay in the insemination of goats

that are observed in oestrus later than 30h after sponge removal is sufficient to

restore normal fertility (Leboeuf, 1994). Leboeuf et al. (2003) inseminated

goats with 100, 75 or 50×106 total frozen-thawed spermatozoa 24h after the

beginning of oestrus behavior. When applying A.I in affixed time in induced

oestrus in goats, interval to oestrus should be considered (Fonseca et al., 2005).

1.4. Gestation or pregnancy Gestation is the period that starts with fertilization and ends with parturition

(the birth process). The average length of the gestation period in the doe is 149-

150 days with considerable variations (Walkden- Brown, 2001). Babiker et al.,

(2010) reported a gestation length in the Sudanese Nubian does of 146 - 152

days, where El Naim, (1979), reported that the gestation length for the

Sudanese Nubian does bearing single kid was 146.5±1.4 days which was

slightly longer than does bearing twins (145.5±1 days), he added that the sex

of the kid and litter size did not significantly affect the gestation length.

Martinez et al. (2005) reported similar gestation length in the Nubian does

(146.5±4.05).

1.4.1. Pregnancy diagnosis

Pregnancy diagnosis is a critical procedure for monitoring goat reproductive

performance, and its an important zoo technical practice necessary to maintain

the high efficiency of a livestock production system (Ortega et al., 1999).



15

various of pregnancy diagnosis methods in goats have evolved over the years

( Kinne, 2006) and these include:

1.4.1.1. Return into oestrus

This is the most commonly used pregnancy test in goats, it involved observation

for signs at 17-21 days post -breeding .Whowever, when inducing oestrus out of

season, in seasonal breeds, the majority of non- pregnant does will not undergo

additional oestrus cycles and will return to seasonal unestrus. Thus, failure to

return to oestrus after breeding, does not indicate pregnancy ( Corteel et al.,

1982 and Smith, 1986).

1.4.1.2. Abdominal palpation

This test like the previous one is cheap and requires no particular equipment. It

is practiced in the second half of pregnancy, it required training and its accuracy

depends very much on the operator. It consists of the presence of the foetus by

forcing it to move in the foetal fluids and bownce against the hand close to the

abdominal wall (Linoel, 1990).

1.4.1.3 Biochemical techniques

1.4.1.3.1. Pregnancy Associated Glycoprotein (PAG):

Pregnancy Associated Glycoprotein (PAG) is one of the proteins from foeto-

placental origin that can be detected in the peripheral circulation of pregnant

animal ( Zoli et al, 1992) and is found in the peripheral circulation of pregnant

dairy and Beef Cows ( Zoli et a.l, 1992), in goats (Benitez and Oritz, 1992 ;

( Ranilla et al.,1997) and in ewes (Ranillia et al.,1994). The level of this

hormone is affected by the breed ( Guilbault et al.,1991), the foetal number

(Benitez and Oritz., 1992) and the period of embryo culture in vitro (Ectors et

al., 1996).



16

1.4.1.3.2. Estrone sulfate

The estrone sulfate test, performed on milk or urine, can determine pregnancy.

Between day 40 and 50 days after conception, the level of estrone sulfate

increases substantially and stays increased throughout pregnancy. Abortion,

foetal death, or resorption causes the esterone sulfate level to drop; therefore,

the test also is a useful measure of fetal viability. The placenta in goats

produces estrone sulfate and it can be detected in goat plasma from around 40-

50 days post breeding (Refstal et al., 1991).

1.4.1.3.3. Progesterone (P4)

Progesterone is one of the C21-steroids (MW=314.5) secreted by the corpus

luteum in females during the oestrus cycle, and in much higher amount by the

placenta during pregnancy. It is also secreted in a minor quantity by the adrenal

cortex in both males and females. Majority of circulating progesterone is bound

to albumin and corticosteroid binding globulin (CBG), the bioactive free

hormone represents only 2.5-3 % of the total progesterone. Measurement of

serum progesterone is of a diagnostic value in menstrual disorders and

infertility in Women. Measurement of progesterone in the first 10 weeks of

gestation, have been suggested in the diagnostic and treatment of patients with

threatened abortion and ectopic pregnancy (IAEA 1984).

Hafez and Hafez, (2000), listed the functions performed by progesterone as

follows:

1. Prepair the endomentrium for implementation and maintenance of pregnancy

by increasing activity of secretory glands in the endometrium and by inhibiting

the motility of the myometrium.

2. Act synergistically with oestrogens to induce behavioral oesturs.

3. Develops the secretory tissue (alveoli) of mammary glands.

4. Inhibits oestrus and the ovulatory surge of LH at high levels.



17

Progesterone is critical for maintenance of pregnancy ( Vasconcelos et al.,

2003). It is synthesized from cholesterol via pregnenolone, then rapidly

metabolized to pregnanediol, for the most part, in the liver. The ovary and

placenta are the major production sites, but a small amount is also synthesized

by the adrenal cortex in both males and females (Thorburn and Schneider,

1972).

Marked differences exit in the level of blood/milk progesterone between

pregnant and non-pregnant females from day 18-19 in the ewe and from day 21-

22 in the doe after fertilization. The assessment of progesterone at this time is a

useful method for detecting non-pregnancy. Measurement of concentrations of

progesterone 19-23 days post-breeding in blood and milk is also a method

for diagnosing pregnancy with high accuracy (Thibier et al.,1982; Murray and

New stead, 1988). Accuracy of diagnosing pregnancy and non-pregnancy was

85.7% and 100% respectively ( Heap and Holdworth, 1981).

Progesterone concentration in milk in does, generally, reflects plasma

concentration, however, concentration of progesterone in milk is higher

(Thibier et al., 1982; Murray and Newstead, 1988). Milk progesterone

concentration above 10ng/ml between 22-26 days after breeding was classified

as positive pregnancy. Milk progesterone concentration varies from day to day

and also with the type of milk sample obtained (Bretzlaff et al., 1989). The

same author reported that plasma concentration of progesterone tends to be

more accurate than milk. Several conditions like hydrometra, pyometra early

embryonic death may extend the luteal life span and give false positive results

(Ishwar, 1995).

The progesterone hormone, When used as an early pregnancy test (18-19 days

after insemination in ewes and 21-22 days in does), the over all accuracy of this

test is higher than 90%. Almost all females diagnosed non-pregnant



18

(prog.<1ng/ml of blood plasma) do not lamb (>98%), while only 75 to 85% of

those presumed pregnant give birth to young. Thus, progesterone estimation at

this stage of pregnancy can be considered as a non-pregnancy test. And at a

late pregnancy (after 19 days in ewes and after 21 days in does), progesterone

measurement can also be used to detect pregnancy. In this case, how ever, the

accuracy depends mainly on sampling frequency as it is impossible to

differentiate between pregnant and cycling females bearing an active corpus

luteum.(FAO, 1990).

Progesterone concentration in serum, plasma or milk can be used as an aid for

monitoring the onset of puberty, postpartum ovarian activity, oestrus cycles and

pregnancy in domestic animals (IAEA, 1999).

1.4.1.3.3.1. Sites of progesterone production

The progesterone is produced by both the corpus luteum and the placenta. Cows

and Sows are dependant on the corpus luteum as a source of progesterone for

most of the gestation period. Progesterone declines during the last half of

pregnancy in these species (Joe, 2000). In both goats and sheep, pregnancy is

maintained by progesterone. Progesterone in ewe is synthesizes by the placenta

in the last–third of pregnancy. However, in goats the sole source of P4

throughout pregnancy is the corpus luteum (Thorburn and Schneider, 1972) and

ovarectomy or stress which upsets the function of the corpus luteum will cause

abortion (Walkden- Brown and Bocquier, 2000).

Thus it is concluded that the integrity of pituitary and the corpus luteum was

essential for the maintenance of pregnancy in the goat and that placenta of goat

did not produce in a sufficient amount to prevent abortion. This conclusion

received support from the findings of Linzell and Heap (1968), who found no

net production of progesterone by the foeto-placental unit of an anaesthetized



19

goat. The rise in the progesterone level in pregnant goat is synthesis by Caprine

placental lactogen (Currie et al, 1977; 1990).

1.4.1.3.3.2. Analysis of the progesterone

One reproductive hormone, progesterone, has been found to be of a significant

clinical value in females of most domestic species and, in fact, overall its

analysis gives the most useful information as to the reproductive status of

animals. While blood has being the usual medium for hormone analysis, milk,

urine, saliva and even faeces are also sources for gaining useful endocrine

information. The later tissues have the advantages in certain situations of being

easier to collect and, at very least, they allow one to avoid the use of

venipuncture (Mats, 2000).

1.4.1.3.3.3. Advantages of the progesterone analysis

Beside its use as an early test for pregnancy diagnosis, progesterone analysis

can also be used for the retrospective confirmation of the absence (or presence)

of a CL at the time of insemination in cattle and this can be a useful tool when

herds with fertility problems are encountered (Karg, et al, 1976; Oltner and

Edqvist, 1982; Garcia and Edqvist, 1990).

A potential important area for the diagnostic use of progesterone analysis

concerns the elucidation of clinical syndromes in the postpartum period in

which cows fail to show sexual receptivity for extended periods of time (Booth,

1988; Sprecher et al., 1990), hence animals which have re-established ovarian

activity can often be distinguished from those which have ovarian inactivity;

elevated progesterone values indicate significant ovarian inactivity is present.

The same author found that progesterone determinations could also be used to

verify the presence of luteal tissue in conjunction with endometritis/ pyometra

and ovarian cysts. Both conditions should respond to prostaglandin therapy,



20

therefore, the use of progesterone analysis would be useful in establishing the

therapeutic response.

1.4.1.3.3.4. Radioimmunoassay for measuring progesterone concentrations:

There are currently several rapid and inexpensive commercial kits available for

use on milk or serum. Results could be qualitative (based on colour changes ) or

quantitative ( progesterone in ng/ml) depending upon the kit (IAEA,1999).

Early and accurate pregnancy diagnosis can be a useful tool for monitoring the

goats reproductive performance (Zoli et al., 1995). Thus enhancing the

reproductive efficiency of flocks.

It is important that clinical endocrinology laboratories understand and have

experience with their assay systems in relation to particular clinical syndromes.

For example, the actual concentration of progesterone during the follicular

phase of the oestrus cycle of domestic animals is approximately 100 pg/ml (318

pmol/liter), certainly not greater than 200 pg/ml (636 pmol/liter) plasma. Some

laboratories, however, reported basal values of 1-2 ng/ml (3.2- 6.4 nmol/liter)

for progesterone (Mats, 2000).

Radioimmunoassays for measuring either progesterone or proteins from

placental and foetal origin have provided early and practical methods for

monitoring pregnancy in farm animals. Progesterone assays accurately detect

non-pregnancy in early gestation (Murray and Newstead, 1988 ; Gonzalez et al.,

(1999); Boscos et al., 2003), the method is based on the competition between

unlabelled progesterone in plasma, serum or milk sample and a fixed quantity

of I125- labeled progesterone for a limited number of binding sites on

progesterone specific antibody immobilized ( coated) on the internal walls of

test tubes ( solid phase). The proportion of the I125 labeled progesterone bound

to the antibody is inversely related to the concentration of the progesterone

found in the tested sample (IAEA, 1999).



21

Serum was used for progesterone testing rather than milk, as serum was readily

available from all animals studied. Some authors suggested that milk

progesterone kits marketed for cattle are unsatisfactory for goat milk since the

goat udder metabolizes progesterone to pregnanedione resulting in false-

negative tests (Holdsworth et al., 1983). So, the ability to utilized serum

samples would increase the usefulness of progesterone test as all goats could be

tested rather than only those which are lactating ( Fleming et al., 1990). Milk

progesterone concentrations are much higher than serum progesterone, and vary

from day to day, according to type of milk sample (foremilk, midmilk, and

strippings) and with production levels in goats (Thibier et al., 1981). A recent

report of the use of a progesterone kit on milk samples in goats indicated

indeterminate progesterone levels in 11.2% of samples while plasma samples

were more accurate on monitoring ovarian activity (Bretzlaff, 1989).

1.4.1.3.3.5. Progesterone profile in oestrus cycle

Determination of hormonal control of oestrus cycle in goats could be based

mainly on plasma progesterone variations (Heap and Linzell,1966; Jones and

kinfton,1977). The circulating progesterone levels, which are characteristically

low during the follicular phase, increase sharply during the luteal phase ,

reaching maximum at 5-10 days after the mid cycle LH peak. Unless

pregnancy occurs, steep decline to follicular levels sets in a bout 4 days before

the next period. During the luteal phase, the concentrations could range between

2.6- 5.4 ng/ml in goats (Khadiga et al., 2005),

The mean progesterone concentration is extremely low at the day of oestrus (0.2

ng/ml) and is not significantly different from that found in unoestrus or

overiectomized animals .The concentration increases to a maximum of 4ng/ml

on about day 10 of the cycle, and decreased rapidly during the last 3 days of

the cycle (Thorburn and Schneider, 1972). The change are closely related to the



22

morphological changes in the corpus luteum during the cycle in the doe. It is

evident from this information that the corpus luteum in goat is not functional

until 3-4 days after ovulation ( Bosu et al., 1978), where (Bono et al., 1983)

found that, the mean plasma value of progesterone during oestrus ranged

between 1-4 ng/ml. Between the 8-56 h before the LH ovulatory peak, the

plasma progesterone levels suddenly decreased, remained lower than 1ng/ml for

a bout 6 days and then gradually increased to typically dioestrus values. Plasma

progesterone drop occurred 24-48 h before behavioral oestrus, whose length

varied between 32-40 h.

The progesterone concentration in the blood decreased sharply during

regression of corpus luteum in the non-pregnant doe (one or tow days prior to

oestrus ) and return to a higher levels within four days following oestrus ( Ott et

al., 1980). Leyva-Ocariz, et al. (1995) compared the progesterone profiles of

native and cross-bred goats in a tropical semiarid zone during the oestrus cycle.

The progesterone levels were found to be similar throughout the oestrus cycle

in both breeds, presenting highest values on days 12 and 15 ( 12ng/ml and

10ng/ml, respectively). For both native and cross-bred does, progesterone

concentrations (2.0±1.2ng/ml and 1.8± 0.2ng/ml, respectively) on day 19

indicated that luteal regression had started. During day 0, one, tow and 20

progesterone concentrations were less than 1 ng/ml (o.5±o.2ng/ml). A

significant difference (P<o.o5) between day 4 (2± 0.49ng/ml) and day 2 in

progesterone levels was found, indicating that the luteal phase was established

from day 4 until day 19.These finding were in a greement with blood levels

found previously (Thibier et al., 1982; Bono et al., 1983). As in the ewe

(Hauger et al.,1977), in native and cross-bred goats and under the tropical

conditions described in this study, at the end of luetal phase of the oestrus cycle,



23

circulating concentrations of progesterone decline while estradiol rises, acting

as a primary signal for the preovulatory gonadotropin surge ( Pant et al., 1977).

There is a paucity of information on hormonal control dynamics after

synchronization of oestrus in does. Most studies have been limited to measuring

responses such as time to onset of oestrus and fertility, this was noticed by

Kusina et al. (2000), who conducted an experiment to study the effect of

different progesterone treatments and PGF2a� on oestrus synchoronization and

fertility of Mashona goats treated with intravaginal sponges. The plasma

progesterone concentrations increased sharply following insertion of sponges to

peak at 6-7 ng/ml within 3 to 5 days after sponge insertion. These levels were

maintained thereafter until removal of sponges. On removal of sponges,

concentration fell sharply to basal levels (< 2 ng/ml within 24h). In does treated

with Norgestomate ear implants, plasma progesterone concentration increased

gradually, reaching a peak of 6 ng/ml prior to removal of the implants. Similar

to the response for does treated with sponges, plasma progesterone

concentrations fell sharply after removal of the ear implants. In does treated

with cloprostenol , the progesterone concentrations plummeted to below basal

levels after the second cloprostenol injection.

Variability in the observed increase in circulating progesterone concentrations

can be quite large (Darwash et al., 2001) and is influenced by feed intake and

milk production ( Lopez et al., 2004; Wiltbank et al., 2006; Elmansoury, 2008 ).

When Greyling and Van der Nest, (2000), synchronized oestrus in different

breeds of goats, Indigenous and Boer goats, they found that, the mean serum

progesterone concentrations for the observation period varied between 0.01 and

6.19 ng/ml for Indigenous and 0.01 and 9.05 ng/ml for Boer goat.

Progesterone started to increase from the mean basal value of 0.1 ± 0.03 ng/ml

on day-0 to 3.0 ± 0.9 ng/ml on day-6 of oestrus cycle and reached the peak



24

value of 7.7 ± 0.6 ng/ml on day 12. From day-15, a decline was observed in

progesterone values till the end of the cycle. (Khanum et al., 2008).

1.4.1.3.3.6. Progesterone and pregnancy

Maintenance of pregnancy is dependant largely on a proper hormonal balance

and the disturbance of this balance results in abortion. Gestation stability is

dependant on an interplay between the mother, the placenta, and the concepts

(embryo or foetus). So progesterone has a dominant role in maintenance of

pregnancy, particularly during the early stage. This fact was noticed by (Joe,

2000 and Vasconcelos et al., 2003), who also noticed that high concentrations

of progesterone decrease the tone of myometrium and inhibit myometrial

contractions. The role of progesterone in down-regulating receptor sites for

estradiol and oxytocin in the myometrium, high progesterone will stop cyclic

oestrus by preventing the cyclic release of gonadotropins.

4.1.3.3.7. Progesterone profile during pregnancy

Several reports are available on progesterone concentrations in blood during

early pregnancy in cattle (Pope et al., 1969). A finding of importance is that

progesterone can be measured in the milk of lactating cows and, further, that its

concentration accurately reflects the concurrent blood plasma concentration of

progesterone (Laing and Heap, 1971).

The difference which exists in both blood plasma and milk progesterone

concentrations 19-24 days after a fertile breeding as compared to a non-fertile

breeding has been used as an early pregnancy test (Shemesh et al., 1968;

Robertson and Sarda, 1971). The plasma progesterone concentrations in blood

of pregnant cows at 21 days post breeding is almost always at least 2 ng/ml (6

nmol/liter) and usually 4-8 ng/ml(13-26 nmol/liter), as compared to less than

0.5 ng/ml (1.6 nmol/liter) in the non-pregnant animal at the same time. Elevated



25

progesterone concentrations, however, only reflect the presence of luteal tissue

and are not directly indicative of the presence of a fetus in vitro.

Furthermore, a slight prolongation of luteal activity in a non-pregnant animal

can occur which results in elevated progesterone concentrations at day 21, a

situation in which the analytical progesterone result would be positive and the

animal would be falsely considered pregnant (Mats, 2000). The same author

add that, the accuracy of the forecast for pregnancy (positive forecast) thus is

often lower than desirable, in most cases ranging between 75 and 90 %. The

negative forecast, however, is more accurate since cows having low

progesterone concentrations in milk or blood 21 days post breeding will almost

always not be pregnant. The accuracy of the positive forecast can be increased

if progesterone analyses are also carried out on samples obtained at the time of

insemination in order to eliminate animals inseminated during the luteal phase

of the oestrus cycle. Cows inseminated during the luteal phase will be in the

luteal phase 21 days later with the animal being falsely considered pregnant.

The plasma progesterone concentrations during early pregnancy in goats (2.5-

3.5 ng/ml) was similar to luteal phase value and remained steady from day 8 to

day 60, between day 60-70 there was a secondary increase in progesterone

concentrations which was maintained in this increase levels (4.5 - 5.5 ng/ml)

until just before parturition ( in twin bearing animals, the secondary increase

was greater). The progesterone concentration decrease rapidly at 1-2 days

preceding parturition, but the concentration was still quite high on the day of

parturition (1.25 ng/ml). (Thorburn and Schnider, 1972).This results were

reported also by Irving et al. (I972) and Humblot et al. (1990). Who

established no trends for a continuous increase or decrease of mean

progesterone concentrations throughout pregnancy.



26

Sousa et al. (1999) studied the progesterone profile in Coninde and Moxoto

goats. They found the following results, during the first week of pregnancy,

the progesterone concentrations were (0.23± 0.07 ng/ml and 0.13±0.03 ng/ml,

respectively). Thereafter, progesterone levels increased significantly ( P<0.05)

in week three (3.35± 0.85 ng/ml and 4.36 ± 0.6 ng/ml), remaining high until

week 19 ( 7.76± 1.33 ng/ml and 5.94 ± 0.90 ng/ml) and decreasing ( P< 0.05 )

just before parturition 3.98± 1.51 ng/ml and 3.45± 0.03ng/ml i.e during

pregnancy mean weekly progesterone concentration varied significantly among

the animals and periods of pregnancy (P<0.01), but they do not affected by

breed or number of fetuses. Llewlyn et al., (1987 and 1992), reported that, the

plasma progesterone concentration rise significantly from 0.91 ±0.51 ng/ml on

day 40 to 5.±o.5 day 60 and remain higher up to 6 ng/ml until 5 days before

kidding. These values in pregnancy is typical to luteal phase in non-pregnant

goats. Those authors also found that, pseudo-pregnant does have similar

progesterone profiles to pregnant does first 80 days, but the rise around day 35

to 40 was not significant and the progesterone concentration returned gradually

to basal level after day one. In indigenous Small East African goats, the mean

plasma progesterone concentration ranged from 2.6 to 10.8 ng/ml from

conception to the mid gestation (Kanuya et al., 2000). After conception, higher

progesterone levels (4.3–11.0 ng/ml) were maintained throughout the gestation

period, and declined rapidly during 19 days pre-partum, reaching 0.8-1.0 ng/ml

on the day of parturition. Zarkawi and Al-Masri, (2002) found that

progesterone concentration in the serum started from basal levels at mating and

become elevated during pregnancy, until a peak level of 13.84±3.77 ng/ml is

reached on day 115.4 ± 23.4 after mating. The concentration drop sharply after

kidding to an average of 0.29 ± 0.25 ng/ml. The mean maximum

progesterone level during gestation recorded by ( Khanum et al., 2008) was



27

10.9 ± 1.3 ng/ml, while Elmansoury, (2008), when he studying the progesterone

profile in pregnant Sudanese Nubian goats, stated that the highest mean of the

monthly progesterone concentration reach up to 17.54±7.60 nmol/L in the first

month, 15.72±4.65 nmol/L in the second month, 16.83±4.44 nmol/L in the

third month, 24.48±7.87 nmol/L in the fourth month and 22.95±7.59 nmol/L

in the fifth month without significant difference (P>0.05). The progesterone

level began at 1.34 ng/ml and then elevated to reach 13.82 ng/ml as a maximum

level and remained elevated above 1 ng/ml until the end of the gestation period

at parturition, this notification was mentioned by Elmoubark, (2010), working

on Sudanese Desert goats. He found that the mean serum progesterone

concentration during pregnancy was 5.93±3.27, 5.51±3.23 and 5.47±2.93 ng/ml

for the different treatment groups. In the breeding season there was a significant

increase (P<0.001) of progesterone concentration in does, (2.85±0.15 ng/ml)

than out side the breeding season (2.37±0.13 ng/ml) and a relevant significance

elevated (P<0.05) level of progesterone in pregnant (3.75±0.17 ng/ml) than in

non-pregnant does (2.47±0.10 ng/ml). This observation was noticed by Al-

Sobaiyl, (2010), on a study about breeding season and pregnancy effect in

Aradi goats.

1.4.1.3.3.8. Relationship between Progesterone and litter size:

The level of progesterone from placental lactogen depend on the number of

faeti (Hayden et al., 1980). Contrast to this, Humblots et al., (1990) found no

relation between the progesterone and the litter size.

Thorburn and Shnieder, (1972), had followed the progesterone concentrations

in six goats, three of which were carrying single fetuses and the other three

carrying twins. The increase in the progesterone concentrations after mating

was the same as that seen in the first part of a normal oestrus cycle. However,

there was no rapid decline in peripheral Progesterone concentrations on day



28

19 as was seen in the non-pregnant animals. In animals carrying single fetuses,

the progesterone concentrations remained at a reasonably steady level (2.5-3.5

ng/ml) from day 8 until about 60; then rose to around 4.5-5.5 ng/ml during the

second part of pregnancy, declining just before partuition. In all the twin-

bearing animals there was a gradual decrease in progesterone concentration

from day 12 until about day 40, a similar decrease was also seen in one animal

carrying a single foetus. The subsequent increase in the progesterone

concentration at about day 60 was greater than in animals carrying single foetus.

In two of the twin-bearing animals, the progesterone concentration decreased

again about day 100 and was then maintained at a similar level to that found in

animals carrying single foetus. Those values in the plasma progesterone

concentrations in does carrying single or twin foetuses are comparable to those

reported for temperate (Chemineau et al, 1982) and other tropical breeds of

goats (Ezzo and Shalably, 1990).

The higher progesterone concentrations in blood plasma of does carrying twins

may be associated with different numbers of corpora lutea as previously

suggested by Chemineau et al. (1982). It is also suggested that plasma

progesterone determination during the fourth to fifth week of gestation can be

used to predict the number of fetus(es) in the Small East African does (Kanuya

et al., 2000). However, Chauhan, et al, (1991), noticed a low accuracy for

predicting multiple pregnancies in goats by use of serum progesterone

concentration. On the other hand successful prediction of the litter size using

progesterone assay in ewes at about 100 days of gestation has being reported

(West, 1986; Schneider and Halford, 1996).

In another experiment studied by Manalu et al. (1997), the average

progesterone concentrations during the last two months of gestation period in

single or twin-bearing does markedly increased (P<o.o1) by 1302 and 2591 %,



29

respectively, as compared to those non-pregnant does (5.791 and 11.113 vs.

0.413 ng/ml). Serum progesterone concentrations increased by 92.4 % as fetal

number increased from single to twins and the average progesterone

concentrations of aborted does had decreased to a very low level, but still

higher than those of non-pregnant does, at least seven weeks prior to abortion

date (1.691 ng/ml for aborted vs. 0.413 ng/ml for non-pregnant does). Where,

Sousa et al. (1999), concluded that progesterone concentrations during

pregnancy were not different between single or multiple pregnancies. Similarly,

Jarrel and Dziuk, (1991) reported that the number of corpora lutea of goats uses

does not influence progesterone concentrations after day 30 of pregnancy in

goats.



30

Chapter Two

Materials And Methods 2.1. Study Area

This study was carried out in the Goat Breed Improvement Project at Helat

Kuku in Eastern Nile Province, Between longitudes E 32 and 33 and latitudes

15 and 36 N and altitude of 380 m above sea level.

2.2. Climate

Details of the climatic conditions of the area have been described below Table( 1). Meteorological data during experimental period (July 2000- January 2001) Season

Temperature (ºc) Relative Humidity Rain fall Day Length Max Min % (mm) (h)

Autumn Winter Summer

39.2 25.2 38 16.98 12.26 32.4 16.9 27 0 11.25 41.1 25 18 0 12.34

*Data obtained from the meteorological Station of Khartoum airport. ` 2.3. Experimental Animals

Sixty healthy Sudanese Nubian does were selected from the total flock of the

project. They were 2.5-5 years old. These does were then subdivided into six

groups each of ten goats. The animals were proved to be empty.

2.4. Husbandry

2.4.1. Housing



31

The animals were housed in an open side-shade pens. The roof was 3.0 meter

high and was made of corrugated and galvanized iron-sheets. The

floor was 1.0 meter above the ground level and was made of concrete (plate).

The selected females were housed into separate pens each was allowed

8.5×0.5m floor area per goat. Each pen was provided with water and feed

troughs of 0.3×0.1m/ animal. The house was well ventilated.

2.4.2. Feeding: -

The animals were fed a concentrate ration which was fed at the rate of 0.5kg/

head twice time a day according to Devednra and Mcleory, (1982), and were

allowed free access to salt lick. Green fodder (Alfalfa or Abu 70) was given

once every day. Drinking water was offered add lipitum

Table (2): Composition of Feed, ingredients of the concentrate ration:

Ingredients Percentages (%)

Sorgum 53

Groundnut cakes 5

Wheat bran 29

Salts 2

Ground nut hulls 11

Total 100 Table (3). Concentrate feed analysis DM% N.F.E% C.P% Fiber

(C.F%) Ash% Fat

(E.E)% Moister (M)%

92.7 61.83 14.4 6.8 5.63 4.00 7.3

D.M = Dry matter. N.F.E = Nitrogen free extract. C.P = Protein 2.4.3. Age

Ages of the selected females were estimated by dentition according to the



32

appearance of the permanent incisors described by Mc Nitt (1983). The age

was ranged between 2.5 to 5 years.

2.4.4. Body condition scoring:

Body condition scoring was assessed for both males and females by palpation

of the lumber region immediately behind the rib and above the kidney, the

degree of projection of the spinal transverse processes of the lumber vertebrae

was assessed according to Russel, (1984). the Body condition scores are range

between 3and 5.

2.5. Treatments

2.5.1. Experimental procedure The does were subdivided into six equal groups. Each group was composed of

10 pure Nubian goats. The groups were randomly assigned to one of the

following treatments.

2.5.1.1. Treatment A:

These animals received no treatment and were observed for natural heat and

mated naturally.

2.5.1.2. Treatment B:

In this group oestrous was induced by introduction of vaginal sponges

impregnated with 40 mg Fluorgestone Acetate (FGA), which is a synthetic

progesterone (Cronogest Intervet International B.V., Boxmeer Holland). The

sponges were removed after 12 days and goats were inseminated artificially

after 48 hours post withdrawal of the sponges.

2.5.1.3. Treatment C:

This group received the same treatment as in group (B) plus an injection with

500 i.u Equine Chorionic Gonadotropin (eCG), intervet, UK:PL 1708-4309

POM, two days before removal of the vaginal sponges and artificial

insemination was conducted at the time of sponges removal.



33

2.5.1.4. Treatment D:

This group received the same treatment as in group (C) but the injection with

Equine Chorionic Gonadotropin (eCG) was applied at sponge removal and

artificial insemination was conducted 48 h later.

2.5.1.5. Treatment E:

In this group the does were injected at the time of progestagen sponges

removal with 300-500 i.u of eCG and 0.5 ml of Estrumate, (synthetic

Prostaglandin F2 a�, Cooper, each ml contains 263 µ�g cloprostenol sodium BP

equivalent to 250µ�g). Insemination was performed 48 hours after eCG and

PGF2a� injection.

2.5.1.6. Treatment F: In this group the animals were injected with 0.5 ml prostaglandin PGF2a� at the

time of sponges removal and the insemination was performed 48 hours there

after.

2.5.2. Sponges application:

Sponges were applied and removed as illustrated in the steps below, and the

instruments used were shown in Figure (3).

Table (4 )-Experimental design INSEMENATION TREATMENT ANIMAL

NO GROUPS

Natural Mating

Non 10 A

AI 48 h post removal of sponges

Progesterone - 10 B

AI at removal of sponges

Progesterone +eCG 48h before removal

10 C

AI 48h post removal of sponges

Progesterone +eCG at removal 10 D

AI 48 h post removal of sponges

Progesterone +eCG + PGF2 at removal

10 E

AI 48 h post removal Progesterone + PGF2 at removal

10 F



34

2.5.2.1. placing of the sponge:

-The applicator was thoroughly disinfected with disinfectant between each

application of the sponge.

-Sponge was inserted with the string- attached end fixed at the near end of the

tube.

-Then the plunger was introduced in the tube.

-The doe was immobilized and the tube was introduced into the vagina.

-The plunger was held and the tube was removed back word to release the

sponge. Then the applicator was withdrawn.

2.5.2.2. Removal of the sponge: -

The sponge was removed gently by pulling back word the string.

2.6. Artificial insemination of does:

Does regardless of oestrus occurrence were inseminated artificially 48 hours

post sponge removal (Fixed time insemination). Semen samples were collected

from four pure Sanneen bucks of known fertility. Immediately before the time

of insemination. Artificial vagina for small ruminants (IMVQ 40) was used.

(Figure 1), semen was collected in a graduated test- tube and then placed in a

water bath at 37c� immediately after collection. Semen samples were generally

evaluated for physical characteristics such as volume, colour, consistency, mass

motility, individual motility, concentration and abnormal morphology as

described by (Evans and Maxwell, 1987). Motility and concentration governed

the volume of Tris diluent to be added. The dilution was done to make final

concentration of 5×108 spermatozoa/ml by pipetting the volume of diluent

required and adding it slowly to the semen, which was held in a baby flask.

Then they were gently mixed, and the diluted sample was then evaluated

under the microscope to confirm sperm viability. The does in the treatment



35

Figure :1, Equipment used in A. I



36

groups whether observed in oestrus or not were inseminated at 48-52 hours

after treatment termination. Site of insemination was interacervical or uterine as

long as the cervix could allow the passage of the insemination gun. A volume of

0.5ml fresh diluted semen containing ( 200×106 ) active spermatozoa was used.

Does to be inseminated were restrained with hindquarter raised over a rail, and

the fore limbs on the ground between the assistant’s legs as described by

(Evans and Maxwell, 1987). A lubricated speculum was introduced into the

vagina to visualize the cervix, the semen was deposited as deeply as possible

into the cervical canal or into the uterus ( 1.5-3.0 cm) into the cervix or into the

uterus, then the semen was deposited by pressing the plunger of the

insemination gun. Figure (1). Goats that returned to oestrus were reinseminated

naturally.

2.7. Pregnancy Diagnosis

Pregnancy was diagnosed by using Radio-Immuno-Assay Technique (RIA),

and by calculation of the Non-Return Rate as detected by a vasectomized buck.

2.7.1. Sampling:

Blood samples were collected from the jugular veins in vacutainers tubes then

cetrifugated for 15 minutes at 3000 round/ minute to separate the serum which

was kept in a deep freezer (-20ºc). The samples were collected at the day of

vaginal sponge insertion, mid period of treatment (day 6), day of insemination,

seven days post insemination, fourteen days post insemination, twenty one days

post insemination, and then at 10 days interval till the end of the gestation

period.

2.7.2. Equipments (Figure 3 ):

a. A microlitter pipette with disposable plastic tips.



37

b. A hand held mulit-dispencer instruments (two) with combitips (Eppendrof

4780 pipette) one for the samples and other for the tracer (I125).

c. A foam decantation rack to hold the coated tubes.

d. A ?�Vortex?� mixer

e. A multi-well gamma counter.(Figure 2)

2.7.3. Materials and Reagents:

a. Antibody- coated tubes, Polypropylene tubes coated with progesterone

antibodies.

b. Tracer I125- progesterone labeled with radio active iodine- (I125).

c. External and internal quality control (EQC) and (IQC).

d. Standards of different counts of progesterone concentrations in blood serum,

as shown in table (5).

Table (5): Serum progesterone standards supplied with the FAO/IQEA

radioimmunoassay Kit (in nmol/L and ng/ml)

Standard Nº nmol/L ng/ml

A 0.0 0.0

B 0.3 0.1

C 1.6 0.5

D 6.4 2.0

E 15.9 5.0

F 31.8 10.0

G 63.6 20.0



38

Figure :2, Gamma Counter

Figure :3, Equipment used in RIA



39

2.7.4. Radioimmunoassay Procedures:

The RIA technique was applied as what mentioned in FAO/IAEA assay

protocol version 3:1 (1996).

a. Samples and other assay components were brought at room temperature.

b. The precoated antibody tubes were labeled for total count (TC), standards,

EQC. and IQC (double for each concentration), and samples.

c. Tested samples and standard and QCs were mixed with the vortex.

d. Hundred µL (100µL) of tested serum, standards and QCs were pipette into

the bottom of the pericoated tube using a microlitre pipette with a disposable

tips.

e. 1 ml of the tracer was pipetted to standards, samples and QCs with the

multi-disposable pipette

f. The tubes were covered with aluminum foil and kept in 40C for 3-4 hours.

g. All the tubes except the total count (TC), which should be placed in a

separate rack, were decanted vigorously in appropriate radioactive waste

disposable pool and the rack was kept upside down on an absorbent paper and

allowed to drain for 2-3 mins and again stirked sharply downwards to remove

residual droplets.

h. The radioactivity were counted in all the tubes.

2.7.5. Radioactivity Measurement and Result Calculation

a. The radioactivity in all tubes ( include TC) were count in a gamma counter

for fixed time (60 seconds). The maximum percentage binding (B max) is

calculated by dividing the average counts per minutes (CPM) of two zero

binding tubes (Bo) by average CPM of the two TC tubes and multiplying by 100.

B max = Average CPM of Bo(standard Zero) X 100

Average CPM of TC



40

*B max. is a measure of how well the assay is functioning ( i.e. how well

progesterone binds to the antibody coated onto the tubes).

b. The percent binding values (B/Bo %) for all standers, samples and quality

control tubes (QCs) was calculated by dividing each CPM of these tubes with

that of Bo (Zero standard) tubes and multiplying by 100.

B/B0 = Average CPM of standard/ sample/ QCs × 100

Average CPM of Bo

* B/B0 is the percentage of the standard, samples, or QC bound by the antibody

in comparison with the B max.

c. A logit- log graph paper was used to plot the average percent bound (B/Bo)

on vertical (Y) axis and the progesterone stander concentrations on the

horizontal (X) axis and straight line (best- fit) through the points on the graph

were drawn, the progesterone concentrations of samples and QCs can then be

measured by reading their percent binding values and interpolating from the

standard curve to the progesterone concentrations on the X-axis. In this

experiment all these calculations of the result were achieved by using special

computer program as shown in figure (2).

2.8. Statistical Analysis:

The comparisons of progesterone concentrations between pregnant and non-

pregnant does were achieved by student's t-test, and the significance between

different treatment groups were achieved by One way Analysis of Variance

(ANOVA), Soft ware, SPSS, version 16. A p-value of less than 0.05 was

considered statistically significant. The percentages were compared by using

Chi- square test.



41

Chapter Three RESULT 3.1.Reproductive Performance in Nubian Does after

Synchronization using Progestagens

3.1.1. Non Return Rate (NRR) % Non- return rate percentages after first service were 50%, 50%, 60%, 60%,

50% and 40% in group A, B, C, D, E and F respectively. There were no

significance differences (P>0.05) between the different treatment groups.

table(6).

3.1.2. Conception/Pregnancy Rate (PR) %

3.1.2.1. Pregnancy rate (%) according to progesterone concentration at day

21 post insemination

Pregnancy rate at day 21 post insemination according to progesterone

concentrations were 30% in group B, 40% in groups A and F, and 5o% in

groups C, D and E without significant differences (P>0.05), as in table (6).

3.1.2.2. Percent of does confirmed pregnant by abdominal palpation 90

days post insemination

Pregnancy rate was found to be 40% in the group A, B, E and F without

significant difference ( P>0.05) while it reached 50 in group C and D without

significant difference( P>0.05), as illustrated in table (6).

3.1.3. Kidding Rate (KR) %

In this study group D showed the highest kidding rate (40%) while it was 30%

in the other groups (A, B, C, E and F). No significant differences ( P>0.05)

where found between these groups, as shown in table (6).



42

Table (6): Reproductive performance in Nubian does after synchronizations using Progestagens Synchronizations protocols - Parameters A B C D E F - Number of Does 10 10 10 10 10 10 Non Return Rate (NRR) % 5(50) 5(50) 6(60) 6(60) 5(50) 4 (40) Pregnancy Rate (PR) %, by palpation (Day 90) 4(40) 4(40) 5(50) 5(50) 4(40) 4(40) Pregnancy Rate (PR) %, According to P4 conc. at 4(40) 3(30) 4(40) 5(50) 5(50) 4(40) day 21 post insemination Kidding Rate (KR)% 3(30) 3(30) 3(30) 4(40) 3(30) 3(30) Fecundity 0.7 0.6 0.6 1 0.5 0.4 Twining rate (TR) 100 67 67 100 67 33.3 Litter size 2.3 2.0 2.0 2.5 1.7 1.3 Gestation length / day 142±2.1c 152±1.9a 147.3±1.5 b 149±1.5ab 147.7±0.66b 148.5±2.1b . Values with different superscript within the same row differ significantly (P<0.05) A = Natural oestrus and natural mating. B = Progesterone +AI. 48 hrs, after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after removal

`



43

Table (7): Effect of litter size and sex of concepts on the gestation length of Nubian does

Litter size Sex

Concept

Single Twin Male Female

Gestation length(Days) (means

±SD)

149.09±3.3

149.09±2.2

149.00±3.08

148.75±2.62

*Significance is done at (P< 0.05).



44

3.1.4. Fecundity: Fecundities of Nubian does valued 0.7, 0.6, 0.6, 1 , 0.5 & 0.4 for the (A, B, C,

D, E and F respectively), as in table (6).

3.1.5. Littre size: The litter size for Nubian does was found to be 2.5 in group D, 2.3 in group A,

2 in groups B and C, 1.7 in group E and 1.3 in group F, as in table (6).

3.1.6 .Twining Rate (TR)%: The twining rate was found to be 100% in groups A and D, 67% in groups B,

C, and E, and 33.3% in group F. as shown in table (6)

3.1.7. Gestation Length/ day:

As illustrated in table (6) the gestation lengths in the Nubian goats were

significantly long in-group B and shorter in the group A, but were comparable

in-groups C, D, E, and F. Regardless to the hormonal treatment protocol there

were no differences in gestation length in Nubian does carrying singles

(149.09±3.3 days) or twins (149.09±2.2 days) and no effect of the sex of

concept on the gestation length as shown in table (7).

3.2. Association of Progesterone (P4) Concentrations with fertility

3.2.1. Progesterone (P4) concentrations before treatment for

oestrus synchronization: A large variations on the progesterone concentrations were registered between

the does in each group before application of the vaginal progesterone

impregnated sponges according to the stage of oestrus at sampling. So goats

with low progesterone concentrations (<3.18 nMol/L) were assumed to be in

oestrus, while those with high progesterone concentrations (>3.18 nMol/l) were

assumed to be in luteal phase.



45

Table (8): Progesterone (P4) concentrations before treatment for oestrus synchronization,

in does carried to full term and does not carried to full term (nMol/L):

Synchronizations protocols

Parameters A B C D E F

N0. of does 10 10 10 10 10

10

Does not carried to full term

Mean ±Sd (nMol/l)

-

7.94 ± 8.69A

(7)

18.91± 29.89A

(7)

7.59 ± 11.00A

(6) 12.59 ± 11.69A

(7)

17.49 ± 12.49A

(7)

Does carried to full term Mean ±Sd (nMol/l)

-

15.03± 13.69A

(3)

5,43± 8.72A

(3)

18.39 ± 11.22A

(4)

28.07 ± 10.15A

(3)

13.38 ± 12.18A

(3)

P-value* - 0.34 0.31 0.17 0.08

0.68

Values between brackets are number of does.

*Values with different superscript within the same column differ significantly (P<0.05). A = Natural oestrus and natural mating. B = Progesterone +AI. 48 hrs after removal.

C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after removal



46

Although individual does varied in their progesterone values, yet no significant

differences were found between the means of the groups (P>0.05) .

The mean progesterone concentrations of does carried to full term before the

hormonal application were insignificantly (P>0.05) higher than that of does

that not carried to full term in groups B, D and E while insignificantly (P>0.05)

lower in group C&F.

There were no significance differences (P>0.05) in the progesterone

concentrations between means within the different groups of either does carried

to full term nor Nubian does not carried to full term at the day of

synchronizations, table (8).

3.2.2. Progesterone (P4) concentrations in does carried to full

term & does not carried to full term at mid-period (day 6) of

Synchronizations: The mean progesterone (P4) concentrations during the mid-period (day 6) of

hormonal treatment were always high and this is assumed to be due to the

presence of progesterone impregnated sponges that releases the hormone into

the blood stream thus keeping its concentration at high levels. As shown in

table (9) the mean progesterone concentrations of the does carried to full term

were higher than that of does not carried to full term, for all groups ( except

group B). Group D from the does carried to full term showed the highest P4

concentrations at the mid period (4o.63 nMol/l).

There were no significant differences (P>0.05) in the progesterone

concentrations among means of different treatment groups of either

carried to full term nor Nubian does not carried to full term at the mid-period

(day six) of hormonal applications as in table(9).



47

Table (9): Progesterone (P4) concentrations in does carried to full term & does not carried to full term at mid-period (day 6) of

Synchronization (nMol/L):



N0. of does 10 10 10 10 10

10


Mean ±Sd (nMol/l)

-

20.13 ± 19.77A

(7)

10.89 ± 10.03A

(7)

31.05 ± 28.31A

(6)

20.27 ± 9.33A

(7)

36.09± 18.08A

(7) Does carried to full term

Mean ±Sd (nMol/l)

-

13.72 ± 12.96A

(3)

13.16 ± 11.84A

(3)

40.63± 20.53A

(4)

29.91 ± 27.68A

(3)

37.17± 22.99A

(3)

P-value* - 0.5 0.77 0.55 0.40

0.93


*Values with different superscript within the same column differ significantly (P<0.05). A = Natural oestrus and natural mating. B = Progesterone +AI. 48 hrs after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after removal



48

3.2.3. Progesterone (P4) concentrations in does carried to full

term & does not carried to full term at the day of oestrus/

insemination: As shown in table (10), progesterone concentrations at the day of insemination

or mating were almost low in the does not carried to full term except two does

from group B and one doe in each of groups, C, D, E and F. while they were

consistently low in the does carried to full term (> 3.38 nMol/l).

The mean progesterone concentration in the does carried to full term was

lower than that of the does that not carried to full term. Even the overall P4

values for the does carried to full term in the groups were less than that in does

not carried to full term but without significant difference (P>0.05).

Concerning does that not carried to full term, the mean progesterone

concentration of group A (1.07nMol/l) is the lowest concentration among the

groups and group B showed the highest P4 concentration ( 14.99 nmol/l)

appendix ( ). The rest of the groups were similar in their P4 concentrations. No

significant differences were found between the means of P4 among the does

not carried to full term in the different groups at insemination (P>0.05).

The does carried to full term in group C registered the lowest P4 value at

insemination (0.20 nMol/l).

3.2.4. Progesterone (P4) concentrations on day 7 post-

insemination in does carried to full term and does not carried to

full term: Progesterone concentrations of the does carried to full term seven days post

insemination were always high (>14.08 nMol/l) in all treatment groups, while

most of the does not carried to full term registered low levels of the P4



49

Table (10): Progesterone (P4) concentrations in does carried to full term & does not carried to full term at the day of oestrus/

insemination (nMol/L):



N0. of does 10 10 10 10 10 10


Mean ±Sd (nMol/l)

1.07±0.96A

(7)

1.63 ± 1.31A

(5)

3.97±8.95A

(7)

3.90±4.27A

(6)

3.76±2.85A

(7)

4.33±2.21A

(7)

Does carried to full term

Mean ±Sd (nMol/l)

2.02±0.41A

(3)

1.79±0.92A (3)

0.20±0.15A (3)

1.56±1.02A (4)

2.82±1.08A (3)

1.85±1.06A (3)

P-value* 0.07 0.21 0.31 0.42 0.47 0.37

Values between brackets are number of does. *Values with different superscript within the same column differ significantly (P<0.05). *Tow does in group B with high progesterone concentrations had been excluded from statistical analysis A = Natural oestrus and natural mating. B = Progesterone +AI. 48 hrs after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after removal



50

Table (11): Progesterone (P4) concentrations on day 7 post-insemination in does carried to full term and does not carried to

full term (nMol/L): Synchronizations protocols


N0. of does 10 10 10 10 10

10


Mean ±Sd (nMol/l)

7.01±0.53A

(7)

4.40±6.08A

(7)

22.36±22.86A

(7)

11.91±10.34A

(6)

19.66±19.21A

(7)

4.82±8.16A

(7)


Mean ±Sd (nMol/l)

25.52±6.11B

(3)

24.65±3.00B

(3)

33.11±22.27A

(3)

49.44±22.96B

(4)

28.05±15.92A

(3)

41.98±8.41B

(3)

P-value*

0.01* 0.00* 0.51 0.00* 0.50 0.00*


*Values with different superscript within the same column differ significantly (P<0.05). A = Natural oestrus and natural mating. B = Progesterone +AI. 48 hrs after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after removal



51

concentrations at this day ( some exceptional does showed high P4

concentrations).

The mean progesterone concentrations of does carried to full term seven days

post–insemination in all the treatment groups were higher than that for the non-

pregnant does These variations in the mean P4 concentrations between does

carried to full term and does not carried to full term were significantly different

(P<0.05) in groups A, B, D and F, while the differences were insignificant

(P>0.05) in group C and E (table 11).

The mean progesterone concentrations 7 days post- insemination for the does

carried to full term were lower in group A and B (24.65 and 25.52 nMol/L)

than that in group E, C and D (28.05, 33.11 and 49.44 nMol/L respectively) but

these differences were not significant (P>0.05), where for the does not carried

to full term the mean P4 concentrations on day 7 post-insemination in groups

B, F and A, (4.40, 4.82 and 7.01 nMol/L respectively) were lower than that of

groups D, E and C (11.91, 19.66 and 22.36 nMol/L respectively) but also these

differences were not significant (P>0.05), as illustrated in table (11).

3.2.5. Progesterone (P4) concentrations on day fourteen post-


full term: Table (12) showed that, the mean progesterone concentrations in all treatment

groups fourteen days post insemination were always high, (>23.15 nMol/L

and >15.58 nMol/L) in does carried to full term and does not carried to full

term, respectively.

The mean progesterone concentrations in does carried to full term fourteen

days post–insemination in all the treatment groups were higher than that for the

does not carried to full term. These variations in the mean P4 concentrations



52

Table (12): Progesterone (P4) concentrations on day fourteen post-insemination in does carried to full term and does not

carried to full term(nMol/L):



N0. of does 10 10 10 10

10 10


Mean ±Sd (nMol/l)

18.02±22.25A

(7)

15.58±33.71A

(7)

16.58±20.25A

(7)

25.90±19.55A

(6)

21.36±20.99A

(7)

18.15±15.27A

(7)


Mean ±Sd (nMol/l)

23.15±11.63A (3)

33.71±6.12A (3)

40.84±1.09A (3)

25.83±25.17A (4)

52.27±17.98B (3)

27.47±5.57A (3)

P-value* 0.66 0.14 0.08 0.59

0.04* 0.19

Values between brackets are number of does. *Values with different superscript within the same column differ significantly (P<0.05). A = Natural oestrus and natural mating . B = Progesterone +AI. 48 hrs after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after removal



53

between does carried to full term and does not carried to full term was just at

the level of significance (P=0.05) in group E, while the remaining groups

showed insignificant differences (P>0.05).

The progesterone concentrations fourteen days post- insemination for the does

carried to full term were significantly indifferent (P>0.05) when we compare

the means of the different treatment groups.

The progesterone concentrations fourteen days post- insemination for the does

not carried to full term were significantly indifferent (P>0.05) when we

compare the means of the different treatment groups. 3.2.6. Progesterone (P4) concentrations on day twenty one post-


full term:

Progesterone concentrations of the does carried to full term twenty one days

post insemination were always high (>14.62 nMol/l) in all treatment groups,

while most of the does not carried to full term registered lower levels of the P4

concentrations at this day ( some exceptional does showed elevated P4

concentrations), as illustrated in table (13).

The mean progesterone concentrations in does carried to full term twenty one

days post–insemination in all the treatment groups were higher than that for

does not carried to full term. These variations in the mean P4 concentrations

between the types of does, were significantly different (P<0.05) in all groups

except group A (P>0.05).

The progesterone concentrations twenty one days post- insemination in the

does carried to full term were significantly indifferent (P>0.05) when we

compare the means of the different treatment groups.



54

Table (13): Progesterone (P4) concentrations on day twenty one post-insemination in does carried to full term and does not

carried to full term(nMol/L): Synchronizations protocols


N0. of does 10 10 10 10

10 10


Mean ±Sd (nMol/l)

7.56±15.39A

(7)

1.42±1.64A

(7)

3.67±6.61A

(7)

5.75 ±12.01A

(6)

7.16±10.36 A

(7)

5.83±9.09 A

(7)


Mean ±Sd (nMol/l)

20.23±7.94A (3)

40.82±16.51B (3)

25.30±4.60B (3)

42.14±22.48B (4)

23.11±7.35B (3)

41.28±21.52B (3)

P-value* 0.21 0.00* 0.00* 0.01*

0.03* 0.00*


*Values with different superscript within the same column differ significantly (P<0.05). A = Natural oestrus and natural mating . B = Progesterone +AI. 48,hrs after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after removal



55

3.2.7. Comparisons between progesterone (P4) concentrations of the goats that not carried to term/ returned to oestrus and these which carried to term during synchronization, at insemination and at first 21 days after insemination Before progestagen treatment the mean Progesterone concentration of all does

carried to full term (14.04 nMol/l) were insignificantly (P>0.05) higher than

that in total does that not carried to full term (12.91 nMol/l), as illustrated in

table (14). At mid- period of progestagen treatment the mean progesterone

concentration in all does carried to full term (26.92 nMol/l) were higher than

that in the total does not carried to full term (23.70 nMol/l), but without

significance difference (P>0.05) table (14). The mean progesterone

concentration in all does carried to full term at insemination (3.86 nMol/l) were

higher than that in the total does not carried to full term (1.71 nMol/l), but

without significance difference (P>0.05) table (14). At seven days post

insemination the mean Progesterone concentration of all does carried to full

term (33.81 nMol/L) was significantly higher (P<0.05) than that for the total

goats not carried to term (11.56 nMol/L) table (14). The mean progesterone

concentration of all does carried to full term (35.77nMol/L) was still

significantly higher (P<0.05) than that for all does not carried to full term

(19.31±18.29 nMol/L) fourteen days post insemination table (14). The mean

P4 concentration in does carried to full term (33.36±16.65 nMol/L) was

significantly higher (P<0.05) than that for all does not carried to full term

(5.46 ± 9.79 nMol/L), twenty one days post insemination table (14).



56

Table (14):Comparisons between progesterone (P4) concentrations (nMol/L) of the goats that not carried to term/ returned to oestrus and these which carried to term at a selected sampling periods

Parameters P4 concentration

before progestagen treatment

P4 concentration at mid period (day 6) of progestagen

treatment

P4 concentration at the day of

insemination (AI)

P4 concentration 7 days after

insemination (AI)

P4 concentration 14 days after

insemination (AI)

P4 concentration 21 days after insemination

(AI)


Mean ±Sd (nMol/l)

12.91 ± 5.25A

(34)

23.70 ± 9.98A

(34)

3.86±6.02 A

(32)

11.56±7.89A

(34)

19.31±18.29A

(34)

5.46±9.79 A

(34)


Mean ±Sd (nMol/l)

14.04 ± 9.63A

(16)

26.92 ± 12.90A

(16)

1.71±0.86A

(16)

33.81±9.97B

(16)

35.77±15.91B

(16)

3.36±16.65B

(16)

P-value

0.82

0.67

0.06

0.00*

0.00*

0.00*

*Values with different superscript within the same column differ significantly (P<0.05).

*Values between brackets are number of does *Tow does, from the does that not carried to term at AI, recorded high progesterone concentrations and had been excluded from statistical analysis



57

3.2.8. Progesterone profile during pregnancy, abortion and early embryonic death During the first days of pregnancy the progesterone concentrations increased gradually with different fluctuating

peaks remaining high (>3.18 nMol/L) until just two to three days before parturition, the serum progesterone

profile of the individual Nubian does during pregnancy was illustrated in figures (4--9), where as figure (10--13)

represent the profiles of does with incompletes pregnancies as a result of abortions or early embryonic death.



58



59

Figure (6). Plasma progesterone concentrations (P4) during gestation in group A. in goats carried to full term.



60

**Arrows refer today of parturition.



61

Figure (7). Plasma progesterone concentrations (P4) during gestation in group B. (day 0 = the day of A.I) in goats carried to full term.



62


.



63



64

Figure (8). Plasma progesterone concentrations (P4) during gestation in group C. in goats carried to full term. ** (day 0 = the day of A.I) **Arrows refer today of parturition.



65

Figure (9). Plasma progesterone concentrations (P4) during gestation in group D. in goats carried to full term. ** (day 0 = the day of A.I)



66




67

Figure (10). Plasma progesterone concentrations (P4) during gestation in group E. in goats carried to full term. ** (day 0 = the day of A.I) **Arrows refer today of parturition.



68



69

Figure (11). Plasma progesterone concentrations (P4) during gestation in group F. in goats carried to full term. ** (day 0 = the day of A.I)



70


Figure (12): Progesterone concentrations of a goat no. a2420 from group (A) showing abortion at day 95 post mating.

• Arrows refer to day of insemination and abortion.



71

Figure(13): Progesterone concentrations of a goat no 1204 from group (C) showing early embryonic death at day 40 post A.I.



72

Figure (14): Progesterone concentrations of a goat no 1806 from group (D) showing early embryonic death at day 36 post A.I.



73

Figure (15): Progesterone concentrations of a goat no 644 from group (E) showing early embryonic death

At 36 day and pregnancy after reinsemination

Chapter Four

DISSCUTION 4.1 Non Return Rate:

Treatment with progesterone is known to significantly improve the conception rate in ovulation synchronization

protocols applied to anestrus does. Our findings (40%-60%) were in range with that registered by Muna et al.

(1998), Elmansory (2008) and Babiker et al. (2010) for the Sudanese Nubian goats; Ali (2004) and Elmubark



74

(2010) for Sudanese Desert goats and Motlomelo et al. ( 2002) in Boer and Indigenous goats. The low

conception rate recorded could be attributed to the insemination being executed too late in some goats in the flocks

or in early stages of the luteal phase at the time of synchronization (Chao et al., 2008; Karikari et al., 2009).

4.2 Pregnancy rate:

In this study, does treated with progesterone with or without eCG showed higher pregnancy rates, compared to the

control group. This probability of pregnancy at first AI in the present study is consistent with other estimates after

using progesterone device mentioned by Motlomelo et al. ( 2002) in Boer and Indigenous goat and was in

agreement with what was recorded by Eppleston et al. (1991); Shalapy et al. (2000) and Yousif (2008) in Nubian

goats, but lower than that obtained by Fonseca et al. (2005). Moreover, progestagen treatment plus eCG injected

48 hours before the removal of progestagen (group D) had better results than when injected at the time of the

removal (group C) and this is similar to what had been described by Leboeuf et al. (2000). While some authors

found that injection of eCG at the time of the progesterone removal had better results (Ritar et al. 1984).

4.3 Kidding rate%

As shown in this study, the injection of eCG, 48 hours before the termination of progestagen treatment, had better

Kidding rate results (40%) than that injected at the time of progestagen removal (30%), this result was similar to

that obtained by Ahmed et al. (1998) that the fertility rate for Sudanese Nubian does treated with progesterone

impregnated sponges plus eCG was 33% and Leboeuf et al. (2000). In contrast Amarantidis et al. (2004) reported

that administration of eCG at termination, or 48h before the termination of intravaginal FGA treatment did not



75

improved fertility. The combination of progesterone plus eCG and PGF2a� had the same effect on kidding rate as

progesterone alone. This confirmed what was reported by Krisher et al. (1993); Hafez and Hafez (2000). Kidding

Rate obtained in this study were less than that reported by Martinez et al.(2005) for the Nubian does (71.4%) and

Kausar et al. (2009) who registed 87.5 Kidding rate % in (Beetal x Dwarf). Kidding rates was less than pregnancy

rates, and that could be due to early embryonic death, or unnoticed abortions. These observations were also

reported by Elmubark, (2001) and Soumia (2002).

4.4. Fecundity rate:

There is a paucity of information concerning fecundity rates in African goat breeds. In the present study the

overall average fecundity rate, across the treatment and control groups, ranged between 0.4 (group F) and 1 (group

D). These values were close to that reported for Northern Kenyan goats, (Hans Meyer et al., 2004), where

fecundity reached 1.18 kids per doe, and lower than that recorded for Nadooshani goats by Bitaraf et al., (2007)

who found the fecundity to be 1.38. Fecundity was not significantly influenced by the oestrous synchronization

methods used in this study, this fact was similarly mentioned by ( Ahmed et al,. 1998) and (Babiker et al., 2010)

when they studied fertility in the same breed.

4.5. Twining rate

There was no significant variations in twining rates between the different treatment groups. The results reveled by

group A, D and E were higher than that reported by Al-Merestani et al. (2003) for Damascus goat and Elmoubark

(2010) for Sudanese Desert Goats, but in agreement with the fact reported by Shahneh et al. (2008) that flushing,



76

especially coupled with eCG treatment increased twining rate, and Kusina et al. (2000), who found that

combination of three hormones (progesterone, PGF2a� and eCG) increased twining rate.

4.6. litter size:

The litter size in this study ranged between 1.3 - 2.5. These values were also reported by Song et al. (2006) who

found the mean litter size in the Korean Native goat was 1.69, Lehloenya et al. (2005) who reported mean litter

size of 2.2 and 2.0, in Boer and Nguni goats, respectively, Imasuen and ikhimioya (2009) who found litter size of

1.75, 2.25 and 2.0 for West African Dwarf Goat treated with 25mg MAP, 5mg MAP and 10mg MAP, respectively.

Martinez et al. (2005) reported a lower litter size for the Nubian does (1.1) when he compared the prolificacy of the

Nubian does with that of Creole and Celtiberians goats.

4.7. Gestation period

The range of gestation periods for the Nubian does found in this study is between 142.25±2.1 and 152±1.9 days. |It

is similar to that found by Devendra and Mcleory (1982), Kudouda (1985) , Jubara (1996) ; Salim et al. (2002) and

Elmubark, (2010) in Sudanese Nubian goats, and Kusina et al. (2000) in Mashona goats. Lehloenya et al. (2005),

in Boer and Nguni goats. Length of gestation period (144.8 days) observed in Dwarf goats by (Amoah et al. 1996)

and Khanum et al. (2007) is comparatively shorter than some other breeds of goat such as 152.1 days in

Toggenburg, 151.1 days in Nubian and 150.5 days in Saanen breed.

The gestation length of does, pregnant with male kids were longer (1-2 days) than in those bearing female kids.

This observation was also recorded by Kudouda (1985) in Sudanese Nubian goats, and (Hafez and Hafez, 2000).



77

4.8. Progesterone (P4) Concentrations before Synchronization ( Day of Hormonal Application):

The serum progesterone concentrations before synchronization were highly variable according to the physiological

state of each doe. Does which were in the luteal phase of oestrus showed higher concentrations, while those in

the follicular phase showed lower concentrations of the hormone. This difference in the progesterone concentration

according to the stage of oestrus was also reported by Leyva-Ocariz et al. (1995), when they compared the

progesterone profiles of native and cross-bred goats in a tropical semiarid zone during the oestrus cycle, on day 0,

one, tow and 20 progesterone concentrations were less than 1 ng/ml (o.5±o.2ng/ml), and presenting highest values

on days 15 and 12 ( 12 ng/ml and 10ng/ml, respectively). For both native and cross-bred does, progesterone

concentrations (2.0±1.2ng/ml and 1.8± 0.2ng/ml, respectively) on day 19 indicated that luteal regression had

started. A significant difference (P<o.o5) between day 4 (2± 0.49ng/ml) and day 2 in progesterone levels was

found, indicating that the luteal phase was established from day 4 until day 19. These finding were in agreement

with progesterone blood levels found previously ( Thibier et al., 1981; Bono et al., 1983). A similar data were

also obtained by Greyling and Van der Nest, (2000), that, the mean serum progesterone concentrations for the

observation period varied between 0.01 and 6.19 ng/ml for Indigenous and 0.01 and 9.05 ng/ml for Boer goat. In

Damascus goats, Khadiga et al. (2005) reported that the progesterone level during luteal phase ranged from 2.6 to

5.4 ng/ml. Khanum et al. (2008) reported that, the Progesterone started to increase from the mean basal value of

0.1 ± 0.03 ng/ml on day-0 to 3.0 ± 0.9 ng/ml on day-6 of oestrus cycle and reached the peak value of 7.7 ± 0.6

ng/ml on day-12. From day-15, a decline was observed in progesterone values till the end of the cycle.



78

4.9. Progesterone (P4) concentrations in does carried to full term & does not carried to full term at mid-

period (day 6) of Synchronizations:

The progesterone concentrations at the mid-period (day 6) of treatment were commonly high, reach up to 36.09

nmol/l (11.35 ng/ml) for the does not carried to full term & 40.63 nmol/l (12.77 ng/ml) for does carried to full

term, without significance differences between the type of does (P>0.05). These results were similar to those

reported by Kusina et al. (2000) who found that the progesterone concentrations increased sharply following

insertion of sponges to peak at 6 to 7 ng/ml within 3 to 5 days after sponges insertion. These levels were

maintained thereafter until removal of sponges. Those results were also obtained in studies in ewes (Kusina et al.,

2000), and in cows ( Walsh et al., 2007., McDougall et al., 2004) that intravaginal treatment of anovulatory

anestrus cows with 1.56g of progesterone has been reported to increase circulating progesterone concentrations to

a peak of 1.2 ng/ml within 24 h and maintain an average serum progesterone concentrations greater than 1 ng/ml

for seven days. In contrast, El-Moristany et al. ( 2000) reported a decrease in the progesterone concentrations

throughout the progesterone sponge treatment (0.25±0.15 nmol/litre and 0.15±0.05 nmol/ litre for MAP and FAG

respectively).

4.10. Progesterone (P4) concentrations in Nubian does carried to full term and does not carried to full term

in the day of oestrus / AI Application:



79

The mean progesterone concentrations at the day of insemination of goats that came in oestrus and show clear

signs were decreased without significant difference between the different treatments, the mean progesterone

concentrations at the day of insemination for the does does carried to full term were decreased below 2.82

nmol/l(0.89 ng/ml) and for the does not carried to full term were decreased below 3.67 nmol/l (1.15 ng/ml) without

significant difference (P>0.05). These results were similar to the basal levels of progesterone in oestrus that

found in other goat breeds ( Zarkawi and Soukouti, 2001; Khadiga et al., 2005; Khanum et al., 2008).

Musadin et al., (1996) reported mean progesterone concentrations during the follicular phase of oestrus cycle as

0.19 and 0.26 ng/ml in Dorset Horn-Malin (DHM) and Malin ewes respectively, but the mean p4 concentation for

group B from non- pregnant does is relatively high (14.99 nmol/l or 4.71 ng/ml) as a result of high readings of the

hormone for tow does in this group (27.29 &56.22 nmol/l i.e 8.58& 17.71 ng/ml) which may be due to a

persistence corpus luteum or assay sensitivity.

4.11. Progesterone (P4) concentrations on day 7,14,21 post-insemination, and during pregnancy in does

carried to full term and does not carried to full term

Progesterone concentration in day 7-post insemination were significantly higher in the does carried to full term

than does not carried to full term (p<0.05). This is due to the presence of the a functional corpus luteum that

secretes progesterone through-out the gestation period to maintain pregnancy. This fact is similar to what was

reported by Kanuya et al. (2000). Some does that does from ones that not carried to full term recorded an elevated

levels of progesterone on day seven after insemination but seems to expose to an early embryonic death. The



80

serum progesterone concentration on day 14 post breeding was higher in both does that carried to full term and that

not does carried to full term, the later assume to be in the luteal phase, these findings were in line with that

reported by (Khadiga et al., 2005), in Damascus goats, where progesterone level during luteal phase ranged from

2.6 to 5.4 ng/ml.

On day 21 post breeding the P4 concentrations of the does carried to full term (20 ng/ml) was higher than that of

does not does carried to full term which assume to return to oestrus. These values were similar to that reported by

Elmobark (2010) and in close agreement with the progesterone profile reported by Zarkawi and Al- Masri

(2002) ,7.24 ± 2.80 ng/ml, and higher than that registered by Selvaraju (2007), 2.5-4.8ng/ml.

In this study and during gestation, higher levels of progesterone were maintained with wide variations between

does like that reported for Dwarf goats (Khanum et al., 2008). Sousa et al., (1999); Humblot et al., (1990);

Thorburn & Schneider., (1972) and Irving et al., (1972) also reported a weekly variation among the does and

during the stage of pregnancy. These authors established that there are no trends for a continuous increase or

decrease of mean progesterone concentrations throughout pregnancy. In indigenous Small East African goats, the

mean plasma progesterone concentration ranged from 2.6 to 10.8 ng/ml from conception to the mid gestation

(Kanuya et al., 2000).

The inconsistency in the magnitude of progesterone in the pregnant does (does carried to full term) is also

reported for the Nubian goats by Elmansoury (2008), in his study the maximum progesterone concentrations was

reached at days 120-135 of pregnancy before it started to decrease significantly in the last tow weeks pre-partum.



81

The same results were also reported by Ezzo and Shalaby, (1990) in sheep and goat. They assumed the decrease as

a result of corpus luteum regression. The progesterone profiles in this experiment is close to the findings of

Llewelyn et al. (1995); Ahmed et al. (1998) and Kanuya et al. (2002), who stated that does exhibited serum

progesterone concentration > 2 ng/ml and maintained these elevated levels during the the gestation period were

considered to be pregnant and were confirmed to be so after kidding.

The overall increase in progesterone levels during gestation and a decline towards the prepartum or parturition,

observed in this study was similar to that reported by (Kadzere et al., 1997; Ozpinar and Firat, 2003; Khanum et

al., 2008). The prepartum decline in the progesterone levels was correlated with the onset of parturition (Laura et

al., 2004). In this study the mean progesterone concentrations were not affected by the litter size (number of foeti ).

These findings were similar to that reported by Jarrell and Dziuk (1991).

4.12. Progesterone profiles in does exposed to abortion or early embryonic death

In this study the kidding rates were less than pregnancy rates which indicate a high loss of embryos. This fact was

also reported by Ahmed et al. (1998), Romano, (2002) and Lu Meng et al. (2008). Johnson et al. (1996) who

observed that two ewes that received 5 mg of progesterone had live fetuses 25 days after mating but returned to

estrus before 40 days of gestation. These researchers indicated that treatment with a low concentration of

progesterone increased the concentration of estradiol and that the low pregnancy rate was attributed to the high

concentration of estradiol. This notification was also suggested by Lu Meng et al., (2008), that the high incidence

of embryonic loss could be attributed to the high concentration of estradiol caused by the low concentration of



82

progesterone and the pregnancy that was supposed to be carried to term was impeded by functionally poor

luteinization. Battye et al., (1988), reported that premature release of PGF2a� from the endometrium might be the

cause of premature regression of the CL in superovulated goats. Hooper et al., (1986) reported that uterine release

of PGF2a� was associated with the secretion of oxytocin by the ovary which controlled luteal regression. It was

suggested that the high embryo loss after synchronization treatment and mating was caused by a functionally poor

CL bringing low doses of progesterone. Further study should be conducted using other methods such as high doses

of progesterone to improve the reproductive performance of goats.



83

Conclusion and Recommendation

1. All the hormonal treatments employed in this study, successfully induced and synchronized oestrus in the Nubian does.

2. Progesterone concentrations during synchronizations were not affected by the different

hormonal methods used to induce oestrus.

3. No significance differences in the progesterone concentrations between goats carried to full term and goats not carried to full term before progesterone impregnated sponges treatment, at the mid-period of sponges treatment and at inseminations. But there is a significant differences in the progesterone concentrations between goats carried to full term and goats not carried to



84

full term seven days post insemination, fourteen days post insemination and at day twenty one post insemination. So blood samples taken at these days can play very important role in controlled breeding programs.

4. Litter size and sex of conceptus had no effect on progesterone profile during pregnancy.

5. Progesterone analysis after artificial insemination proved to be a useful tool for fertility

managements and efficient in early detection of pregnancy, early embryonic loses and abortion.

6. A sustained use of progesterone radioimmunoassay technique in conjunction with A.I.

programs is highly recommended to improve reproductive efficiency in goats.

7. More studies can be establish to know and eradicate causes of infertility in goats in Sudan. Chapter Five

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APPENDIX

Table (8): Progesterone (P4) concentrations before treatment for oestrus synchronization,

In does carried to full term and does not carried to full term (nMol/L):

Synchronizations protocols . Parameters A B C D E F . Total (Mean±Sd) No of does 10 10 10 10 10 10 Does not carried to Full term - 3.66 0.2 7 24.41 0.00 16.66 26.58 23.26 0.76 11.52 19.51 1.89 26.40 0.00 25.57 34.79 6.98 0.59 0.00 21.37 0.83 9.88 0.00 1.61 0.27 2.86 2.02 81.44 18.76 3.52 18.72 4.56 0.57 25.96 29.12 Mean±Sd - 7.94±8.69 18.93±29.89 7.59±11.00 12.59±11.67 17.49±12.49 12.91±5.25 Does carried to Full term - 6.63 15.50 22.45 23.21 0.86 30.83 0.39 23.42 21.27 15.77 7.63 0.40 25.97 39.74 25.00 1.71 Mean ±Sd - 15.0313.69 5.43±8.72 18.39±11.22 28.07±10.15 13.38±12.18 14.04±9.63 - - P-value - 0.34 0 .31 0.17 0.08 0.68 0.82 - Values are significance al level (P<0.05). , A = Natural oestrus and natural mating. B = Progesterone +AI. 48 hrs after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after



120

Table (9): Progesterone (P4) concentrations in does carried to full term & does not carried to full term

at mid-period (day 6) of synchronization (nMol/L):

Synchronizations protocols Parameters A B C D E F Total (M±SD) No of does 10 10 10 10 10 10 Does not carried to Full term - 4.00 1.73 9.94 10.54 29.71 11.81 11.71 5.42 35.52 56.15 23.34 10.95 38.80 28.05 60.00 - 60.00 - 11.94 18.25 16.16 24.53 0.00 40.17 20.70 19.84 0.51 28.10 80.00 19.92 24.00 . 16.68 12.86 - 8.88 47.48 Mean±Sd 20.13±19.77 10.89±10.03 31.05±28.31 20.27±9.33 36.09±18.08 23.70±9.98 Does carried to Full term - 26.23 24.20 53.55 24.21 14.02 0.36 14.62 61.20 5.52 60.00 14.58 0.66 31.39 60.00 37.50 - - 16.37 - - Mean±Sd 13.72±12..96 13.16± 11.84 40.63±20.53 29.91±27.68 37.17±22.99 26.92±12.90 P-value - 0.5 0.77 0.55 0.40 0.93 0.67 - Values are significance al level (P<0.05). , A = Natural oestrus and natural mating. B = Progesterone +AI. 48 hrs, after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after removal



121

Table (10): Progesterone (P4) concentrations in does carried to full term & does not carried

to full term at the day of oestrus/ insemination (nMol/L):

Synchronizations protocols . Parameters A B C D E F Total(M±Sd). No of does 10 10 10 10 10 10 Does not carried to Full term 1.30 3.21 1.00 1.40 9.07 1.14 2.12 27.29 1.57 1.11 1.10 3.56 0.23 0.80 0.00 12.15 1.57 4.74 2.24 1.66 0.39 1.49 4.99. 0.00 0.40 - 0.08 4.89 3.99 22.90 - 56.22 2 4,24 2.37 1.28 0.00 0.11 1.67 0.51 - 4.41 1.42 Mean±Sd 1.07±0.96 14.99±22.64 3.97±8.95 3.90±4.27 3.76±2.85 3.33±2.21 5.16±4.93 Does carried to Full term 2.27 1.01 0.37 0.18 3.51 2.92 2.24 1.55 0.12 1.56 3.38 1.83 1.55 2.80 0.11 1.88 1.58 0.80 2.61 Mean± Sd 2.02±0.41 1.79±0.92 0.20±0.15 1.56±1.02 2.82±1.08 1.85±1.06 1.71±0.86 . P-value 0.07 0.21 0.31 0.42 0.47 0.37 0.00 - Values are significance al level (P<0.05)., A = Natural oestrus and natural mating. B = Progesterone +AI. 48 hrs after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after remov



122

Table (11): Progesterone (P4) concentrations on day 7 post-insemination in does carried to full term and does not carried to full term (nMol/L):

Synchronizations protocols .

Parameters A B C D E F Total(M±SD). No of does 10 10 10 10 10 10 Does not carried to Full term 0.00 6.80 2.51 28.17 60.00 1.15 23.00 - 41.28 - 8.85 4.74 0.00 0.00 1.69 3.00 5.55 0.00 5.38 15.80 60.00 15.81 22.03 22.90 1.52 0.91 33.93 4.61 23.99 0.00 1.06 1.72 4.92 7.94 7.51 3.56 18.11 1.20 12.18 9.69 1.42 Mean±Sd 7.01±0.53 4.40±6.08 22.36±22.86 11.91±10.34 19.66±19.21 4.82±8.16 11.56±7.89 . . Does carried to Full term 25.64 27.99 58.54 54.09 45.38 33.25 31.57 22.20 17.08 80.00 14.08 42.72 19.35 23.75 23.71 30.06 24.69 50.00 33.61 Mean± Sd 25.52±6.11 24.65±3.00 33.11±22.27 49.44±22.96 28.05±15.92 41.98±8.41 33.81±9.97 . P-value 0.01 0.00 0.51 0.01 0.50 0.00 0.00 Values are significance al level (P<0.05). A = Natural oestrus and natural mating. B = Progesterone +AI. 48 hrs after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after removal



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Table (12): Progesterone (P4) concentrations on day fourteen post-insemination in does carried to full term and does not carried to full term(nMol/L):


Parameters A B C D E F . Total (Mean±Sd) No of does 10 10 10 10 10 10 Does not carried to Full term 8.08 3.41 14.98 - 55.93 26.57 60.00 8.99 23.71 24.96 5..01 0.00 21.22 - 1.69 51.90 27.53 0.00 - - 12.18 2.21 1.49 25.04 3.25 34.33 58.54 12.22 0.88 38.08 1.26 - 0.45 38.16 19.33 29.31 13.5 - 4.51 39.37 8.08 Mean±Sd 18.02±22.25 15.58±33.71 16.58±20.25 25.90±19.55 21.36±20.99 18.15±15.27 19.31±18.29 . . Does carried to Full term 32.52 29.47 39.84 14.33 73.64 33.25 26.80 30.93 39.63 69.96 40.94 22.14 10.14 40.72 43.06 20.56 42.88 27.02 42.45 Mean± Sd 23.15±11.63 33.71±6.12 40.84±1.09 25.83±25.17 52.27±17.98 27.47±5.57 35.77±15.91 - P-value 0..66 0.14 0.08 0.59 0.04 0.19 0.00 - Values are significance al level (P<0.05). A = Natural oestrus and natural mating. B = Progesterone +AI. 48 hrs after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after removal



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Table (13): Progesterone (P4) concentrations on day twenty one post-insemination in does carried to full term and does not carried to full term(nMol/L):


Parameters A B C D E F . Total (Mean±Sd) No of does 10 10 10 10 10 10 Does not carried to Full term 0.00 0.00 0.37 0.71 0.02 10.57 0.00 2.89 17.86 0.51 9.11 3.80 1.55 0.00- 0.09 0.81 29.59 0.62 1.76 - 0.25 2.21 1.87 24.29 - 2.80 0.93 0.04 4.08 1.19 3.15 - 6.06 30.23 4.01 0.00 38.87 - 0.12 0.65 0.06 Mean±Sd 7.56±15.39 1.42±1.64 3.67±6.61 5.57 ±12.01 7.16±10.36 5.83±9.09 5.46±9.79 - . Does not carried to Full term 14.62 59.56 30.14 72.94 16.07 65.71 25.85 34.50 20.98 23.22 22.53 32.89 - 28.40 24.77 27.95 30.74 25 .15 44.43 Mean=Sd 20.23±7.94 40.82±16.51 25.30±4.60 42.14±22.48 23.11±7.35 41.28±21.52 33.36±16.65 - P-value 0.21 0.00 0.00 0.01 0.03 0.00 0.00 - Values are significance al level (P<0.05). A = Natural oestrus and natural mating. B = Progesterone +AI. 48 hrs after removal. C= Progesterone + PMSG 48 hrs before removal + AI. At removal D. = Progesterone + PMSG at removal+ AI. 48 hrs after removal E = Progesterone + PMSG & PGF2a� at removal + AI. 48 hrs after removal F = Progesterone + PGF2a� + at removal + AI. 48 hrs after removal



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Documents

AZZA ALI ADAM ABUJE B.V.Sc. University of … · 1.2.2.4 Duration and Dosage of progesterone treatment 10 1.2.2.5 ... Radio Isotopes and Immunology, (CVRL), Souba, Dr. Asha