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1 Copyright is owned by the Author of the thesis. Permission is given for a copy to be downloaded by an individual for the purpose of research and private study only. The thesis may not be reproduced elsewhere without the permission of the Author.

2 STUDIES ON THE USE OF THE CIDR INTRAVAGINAL DEVICE FOR REPRODUCTIVE MANAGEMENT OF DAIRY CATTLE ALBERTO RAUL DICK 1990

3 STUDIES ON THE USE OF THE CIDR INTRAVAGINAL DEVICE FOR REPRODUCTIVE MANAGEMENT OF DAIRY CATTLE A THESIS PRESENTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF PHILOSOPHY AT MASSEY UNIVERSITY ALBERTO RAUL DICK December, 1990

4 GENERAL ABSTRACT This study was undertaken as a series of projects which involved selected studies with the CIDR1 intravaginal device in dairy cattle. In the first project, a CIDR device was inserted at different stages of the oestrous cycle to characterize the dynamic changes of the follicles in the ovaries of cycling cows, the associated changes in plasma progesterone concentrations (PPC), and the effects of progesterone from the device on cycle lengths. The results demonstrated that the progesterone released by the CIDR appeared to enhance the development and maintenance of a dominant follicle. The reproductive responses after device removal were influenced by the follicular population, and by the stage of the follicular wave even in the absence of a corpus luteum (Cl). The PPCs during CIDR insertion or after removal were influenced by the type of animal and the stage of the cycle when treatment was initiated. The stage of the oestrous cycle in which the device was inserted also influenced the average cycle length. In the second project, the tailpainting and raddle technique combined with scoring and re-raddling gave a precise correlation with visual oestrus detection and with patterns in the onset of oestrus in groups of heifers and cows after different synchrony treatments. The average interval to oestrus was concentrated between 30 and 120 h after using different treatments for oestrus synchronization. In the third project, a controlled breeding programme for cows in herds with a daily milk quota was investigated, with a view to improving reproductive performance through the strategic use of the CIDR intravaginal device. The study showed that the mean interval from Planned Start of Mating (PSM) to first insemination was shorter in all treated groups in year-round herds, and treated cows had fewer days open than control. Conception rate to first insemination differed statistically between groups, but not for second inseminations. As a consequence, the mean number of services per conception also differed significantly between groups. Oestrous responses in all farms differed significantly in the treatment group compared with the control group (76% vs 63%; P< 0.001, respectively). The return to oestrus in treated cows was synchronized in 85% of non-pregnant cows on days 22 to 25 after first insemination. The average responses to the two doses of prostaglandin at CIDR removal was similar for half dose vs full dose (72% vs 68%, respectively). However, this management advantage was partly lost because of lower fertility and because of mistakes relating to the interpretation of the tailpainting system which were frequently made by the owners. Controlled internal drug releasing device (Eazi-Breed CIDR""-8, Carter Holt Harvey Plastic Products, Hamilton, N.Z.

5 In the fourth project, the effectiveness of using a milk progesterone test to identify non-pregnant animals and consequently to improve oestrus detection rates was evaluated in a management system for dairy cows involving the use of CIDR devices for controlling return to service intervals. The average percentage of non-pregnant cows inseminated during the second period of artificial insemination (AI) was 63% and varied from 44% to 77.2% among individual herds. In general, the study showed that the identification of non-pregnant animals did not improve the percentage of animals inseminated over the second period of Al. In the last project, the dynamic changes of follicles on the ovaries at two different post-partum periods was characterized as well as the post-treatment response rates in oestrus and ovulation in anoestrous dairy cows. The population of follicles in classes 1 ( < 6 mm), 2 (6 to 9 mm) and 3 (> 9 mm) varied between cows in both post-partum periods, but the average number of follicles did not differ significantly between day 25 (early) and (late) for treatment and control groups. In the late post-partum period, the average number of class 1 follicles increased in the animals of the treated groups (P < 0.05), and when the comparison in treatment groups was made between early and late post-partum period, the average number of class 2 follicles and the total number of follicles were both increased at CIDR removal (P< 0.05; P< 0.01, respectively). The average number of class 1 follicles in the early post-partum period increased significantly in the treatment group irrespective of whether or not animals displayed oestrus or ovulated with CL formation after treatment with CIDR/PMSG. The diameter of the largest unovulated luteinized follicle in treated cows which displayed oestrus and/or ovulated increased significantly during CIDR treatment, and its growth continued after the device was removed. In both post-partum periods, normal and luteinized class 3 follicles were found in non-cycling cows, where some had large normal and others had large luteinized follicles. Only 25% and 33.3% of the treated cows in the early and late post-partum period displayed oestrus, respectively. However, 55% and 50% of the treated and control cows which had not displayed oestrus, actually ovulated and formed a CL in the early post-partum period. From the results of this study, one can conclude that although insertion of a CIDR device into cycling cows synchronizes oestrus, there remains significant variation among animals in the precise time after device removal that oestrus commences. Moreover, some animals have reduced fertility at the synchronized oestrus. To overcome these two limitations on the benefits of synchronization, the duration of treatment programmes based on a progestagen will have to be adjusted to obtain high conception rates, and greater account will have to be taken of follicle wave patterns as a factor influencing precision of synchrony. These considerations apply also to the re-use of CIDRs after insemination, to reduce variation in return to oestrus and to improve fertility in cows which do not conceive to insemination at the first synchronized oestrus. To ensure precise synchrony it is important to emphasize that the most common problem in breeding management in dairy cows is inadequate oestrus detection. The tailpainting system may be less satisfactory in herds with continuous calving patterns. lt shows particular potential for further studies

6 between animals which may not display oestrus, and the associations of the tailpaint and raddle technique in animals with different coats and skeletal conformation. In non-cycling cows, further studies have to be focused on the importance of identifying factors which affect the treatment response, the endocrinological characteristics of these animals, and the interactions of the nutritional status, body condition and ovarian function during the post-partum period. iii

7 ACKNOWLEDGEMENTS This thesis is dedicated to the memory of my father Julian Alberta, who was my best friend and father. Thanks viejo (dad) I am very proud to be your son. My project was unusual in that one of my objectives was to come to New Zealand to work with Or K. L. Macmillan of Ruakura Agricultural Centre, because of his international reputation in dairy cattle reproduction. Since Or Macmillan does not work in a University, the project was set up at Massey University under the joint supervision of Professor R. S. Morris (who has arranged my training and research activities) and Or Macmillan (who has provided the specialist research guidance in reproduction). I therefore wish to express my sincere thanks and appreciation to Or K. L. Macmillan for his continued interest, guidance, support and invaluable advice, not only in the completion of this work, but also in the development of my way of thinking in reproduction. I also wish to express my gratitude to Professor Morris for his invaluable guidance, advice and organizing my research to ensure the completion of this study. I appreciate the help and guidance of Professor Fielden, who was eo-supervisor during the first part of my training here. I also appreciate the rewarding discussions that I had with Professor N. Williamson during the final part of my studies here at Massey. Special gratitude is extended to Mr Chockchai Chaimongkol of Thailand for valued friendship and willing assistance throughout the entire duration of my research in this country. Special thanks is extended to Debbie Love lock, Dirk Pfeiffer, Robert Sanson, Bill Tranter, Eugene Lanada, Brigitte Revol, Kerry Killorn and students of the 1989 fifth year class for their assistance with my experimental work. Also technical and farm staff of the Department of Veterinary Clinical Sciences, the Dairy Cattle Research Unit and Massey N 4 Dairy Unit for their assistance in various ways during my work. Or K. Lapwood and Ms Jane Candy for the advice and assistance with the progesterone and LH assays. Professor S. McCutcheon for my training in cannulation in cattle. Fiona Dickinson for the assistance in com puter programmes and her many helpful suggestions in my thesis preparation. Special thanks are due to the dairy farmers in the Manawatu, for allowing me to work with their cows, in particular Mr and Mrs Johnston, Mr and Mrs Hislop, Mr and Mrs Cowen, Mr and Mrs McCarthy and Mr O'Connor and also for their help to complete my field research.

8 The financial support from the New Zealand government is also acknowledged. All the other people, too numerous to mention, of the Department of Veterinary Clinical Sciences for their friendship and assistance. A special thanks is extended to Emilio Ruz and J. Guillermo Cruz C. for their advice and help with the preparation of my thesis. I have a deep respect for my Kiwi friend Sara Cotterall, who gave not only assistance and support, but also shared her knowledge and experiences with me which has helped make my stay in New Zealand a happy one. A very special thanks is also extended to my dear mother, my Argentinian friends and relatives for their love, their faith in me, and their moral support during my studies here at Massey. Finally, my wife Diana, and children, Martin, Mariana and Laura for always having the patience, tolerance, love and understanding while I com pleted this thesis, without which it would never had been written. V

9 TABLE OF CONTENTS Page General Abstract iii Acknowledgements v Table of Contents xviii List of Figures xx List of Tables xxvi General Introduction CHAPTER 1 : Ovarian Follicular Population Oestrous Response Plasma Progesterone Concentrations and Length of Cycles in Cows Treated with an Intravaginal Device at Different Stages of the Oestrous Cycle Abstract Introduction Literature Review Ultrasonic lmaging and Reproductive Events in Cattle Principles Interaction of the Sound Beam with Tissues Equipment Uses Controlled Breeding in Cattle Progestagens

10 Progestagens-Oestrogen combinations Prostaglandin F2 alpha and its Analogues (PGF) Progestagen-Prostaglandin combinations CIDR-8 Indications Oestrous cycle control in adult cattle Effects of using CIDR-8 after first insemination on pregnancy rate and subsequent synchrony CIDR-device use as a contraceptive Plasma Progesterone Concentrations in Cycling and Ovariectomized (OVX) animals treated with a Progesterone Intravaginal Devices vii Objectives Material and Methods Animals Experimental Protocol Method of use of the CIDR Blood Collection and Analysis Normally cycling (NC) group Ovariectomized (OVX) group Procedure of Jugular Cannulation Material and equipment Procedure Removal of a cannula Ovariectomy Hormone Assays Progesterone

11 viii Luteinizing hormone Ultrasound Examination Validation of ultrasonic measurements of follicular diameter Statistical Analysis Results Animals Pro-oestrous CIDR insertion (PCI) Group Metoestrous CIDR insertion (MCI) Group Early-Dioestrous CIDR insertion (E-DCI) Group Late-Dioestrous CIDR insertion (L-OCI) Group Late-Dioestrous + PGF during CIDR insertion (L- OCI + PGF) Group Corpus Luteum Effects on Follicular Diameter and Number of Follicles Oestrous Cycle and Follicular Status Interactions between Follicle Classes and Interval to Oestrus Interactions between Follicle Class, Ovary and Interval to Oestrus Normally Cycling (NC) Group Ovariectomized (OVX) Group Individual Animal Variation Validation of the Ultrasound Data Discussion Oestrous Cycle and Follicular Status Corpus Luteum Effects on Follicular Diameter and Number of Follicles

12 ix Follicle Class, Ovary and Interval to Oestrus Plasma Progesterone Profiles Plasma LH Profiles Progesterone and Oestrous Cycle Length Validation of the Ultrasound Data Conclusions References CHAPTER 2: The use of Tailpaint and an Aerosol Raddle to monitor Oestrous Behaviour of Animals after different Synchrony Treatments Abstract Introduction Literature Review Oestrous Synchronization in Cattle Prostaglandin F2 alpha and its Analogues (PGF) Progestagens CIDR-8 Indications Synchronization of Heifers for Artificial Insemination using CIDR First experimental series Second experimentat series Third experimental series Detection of oestrus The duration of Oestrus Frequency and Time of Observation

13 Methods of Oestrus Detection Hormonal Changes Changes Associated with Oestrus Electrical resistance of the vaginal mucus Body temperature Oestrous odour Other behaviour Detection of Standing Oestrus other than by Visual Observation Television cameras Vasectomized bulls or testosterone treated teaser cows or steers with chin-ball harness " Other Methods " Oestrus Detection Rate X Objectives Material and Methods Animals Observations of Oestrus and Tailpainting and Raddle Scoring Tailpainting Raddling Experimental Protocol Trial Trial Blood Collection 101 Hormone Assay 101

14 xi Plasma progesterone Milk progesterone Statistical Analysis Results Trial Observations of oestrus and initial tailpaint raddles scores for each individual animal Patterns of onset of oestrus Duration of the oestrus Sexual active group and its composition Tailpaint and raddle and mounting activities 105 Trial Observations of oestrus and initial tailpaint scores for each individual animal Patterns of onset of oestrus Duration of the oestrus Sexual active group and its composition Tailpaint and raddle and mounting activities Progesterone profiles Discussion Tail painting and Raddle Scoring Patterns of Onset of Oestrus Duration of Oestrus

15 Sexual Active Group and its Composition xii Conclusions References CHAPTER 3: Controlled Breeding Management through the strategic use of the CIDR B intravaginal Device for Cows in Herds with a Daily Milk Quota Abstract Introduction Literature Review Studies of Synchronization of Oestrus in Dairy Herd Reproductive Programmes Management System for Synchronization of Oestrus in Cattle utilizing PGF at Various Dose Rates Effect of treatment with Progesterone on Fertility Anoestrum Oestrus Detection Objectives Material and Methods Experimental Design Experimental Procedure Control Group Progesterone Determination Hormone Assay Statistical Analysis

16 Results Herd Reproductive Efficiency xiii Farm A 147 Farm Farm C 147 Farm D 148 Herd reproductive efficiency in "Townmilk" herds Anoestrum in Lactating Dairy Herds Town supply herds Autumn-Spring calving herd Oestrous Response after Synchronization by using CIDR-8, PGF and PMSG Farm A 150 Farm Farm C 151 Farm D 152 General oestrous response after synchronization Oestrous Response and Use of Full and Half Dose of Prostaglandin at CIDR-8 Removal Effect of Treatment with Progesterone on Pregnancy Rate Plasma Progesterone Discussion Herd Reproductive Efficiency Anoestrus in Lactating Dairy Herds Oestrous Response after Synchronization

17 xiv Oestrus Response and Use of Full and Half Dose of PGF at CIDR-8 Removal Plasma Progesterone Conclusions References CHAPTER 4: The use of milk Progesterone Tests to identify Non-pregnant Animals and consequently improve the Oestrus Detection in a Management System for Dairy Herds involving the use of CIDR Devices Abstract Introduction Literature Review Early Detection of Non-pregnant and Pregnant Animals Confirmation and Prediction of Oestrus Identification of Resumption of Ovarian Activity after Calving Monitoring of Reproductive Components associated with Ovarian Disorders and Embryo Mortality Integration of Hormones Assay into Fertility Control Programmes Assay Methods of Measuring Progesterone Principles and Characteristics of Milk Progesterone Tests Enzyme-linked immunoabsorbent assay Latex aglutination test

18 Objective Material and Methods Experimental Design Experimental Procedure Control Group Progesterone Determination Hormone Assay Milk progesterone Plasma progesterone XV Statistical Analysis Results Discussion Conclusions References CHAPTER 5 Ultrasonic Identification of Ovarian Follicles and Response Patterns in Anoestrous Dairy Cows Treated with CIDR Devices and PMSG at two Stages of the Post-partum Period Abstract Introduction Literature Review Post-partum Period Genital tract function Uterine involution Involution process

19 xvi Hypothalamic-pituitary ovarian function Pituitary hormones Adrenal cortex Uterus Mammary gland Effects of steroids on LH and FSH release 238 Progesterone priming and its effect on silent ovulations in the postpartum period Follicle growth pituitary hormones and 240 Follicle maturation and gonadotrophinsecretion before ovulation Anoestrus 242 Ultrasound and Animal Reproduction Objectives Materials and Methods Animals Experimental Protocol Blood Collection and Analysis Treatment groups Control groups Hormone Assay Progesterone Ultrasound Examination

20 xvii Statistical Analysis Results Level of Production and Body Condition Score Follicular Status of the Ovaries at Initial Examination Early post-partum 250 Late post-partum 250 Follicle Number during Treatment Early post-partum 251 Late post-partum 251 Follicle Changes before Oestrus Early post-partum 252 Late post-partum 252 Plasma Progesterone Profiles Early post-partum 253 Late post-partum 253 Oestrous Response and Ovulation after Treatment 253 Early post-partum 253 Late post-partum 254 Discussion Follicular Status of the Ovaries at Initial Examination Follicle Number during CIDR-B Treatment Follicle Changes before Oestrus Plasma Progesterone Profiles

21 xviii Oestrous Response and Ovulation after Treatment 257 Conclusions References Appendix

22 LIST OF FIGURES Page Figure 1.1 Figure 1.2 Figure 1.3 Figure 1.4 Figure 1. 5 Figure 1.6 Figure 1.7 Figure 1.8 Figure 1.9 Figure 2.1 Figure 2. 2 Figure 3.1 Figure 3.2 Properties of a typical longitudinal wave. Each wave has a characteristic wavelength (. ), frequency, and velocity. The amplitude represent the intensity of the sound The various potential acoustic interfaces encountered by the sound beam. The arrows represent pulses of sound CIDR Devices/applicators Timetable of experimental protocol Plasma LH concentrations in cow A from oestrus until + 6 H after oestrus Plasma LH concentrations in cow B from oestrus until + 6 h after oestrus Plasma LH concentrations in cow C from oestrus until h after oestrus Average diameter of largest follicle in cows with or without a corpus luteum (CL) Average plasma progesterone concentrations in ovariectomized cows treated with CIDR-B for 10 days Distribution intervals to onset of oestrus in groups of heifers which received different synchrony treatments Distribution intervals to onset of oestrus and/or first insemination in groups of cows which received different synchrony treatments Basic programme of experimental protocol Percentage of cows in oestrus and inseminated in treatment groups throughout the study period

23 XX Figure 4.1 Figure 4. 2 Figure 4.3 Figure 5.1 Figure 5. 2 Figure 6.1 Basic programme of experimental protocol Non-pregnant and pregnant cows and progesterone in milk by radioimmunoassay (RIA) Non-pregnant and pregnant cows and progesterone in plasma by radioimmunoassay (RIA) Ultrasound images of follicles at various stages of the oestrous cycle Ultrasound image of a luteinized follicle in a noncycling cow at day 14 after treatment with CIDR/PMSG A transrectal real time linear array ultrasound scanner (a), equipped with a 5.0 MHz rectal probe (b), and a video recorder (c)

24 LIST OF TABLES Page Table 1.1 Average number of follicles in each class (class 1, < 6 mm; class 2, 6-9 mm and class 3, > 9 mm) for cows at different stages of the oestrous cycle during the CIDR-8 treatment period Table 1.2 Average plasma progesterone concentrations (PPC[Mean (SEM); ng/ml]) at the time of CIDR insertion (TD 0) and at selected intervals during treatment (TD h, TO 3, TO 10 and TO h) in ovariectomized (OVX) and cycling cows Table 1.3 Table 1.4 Table 1.5 Table 1.6 Table 1. 7 Oestrus cycle length in cows treated with a CIOR_8 for 10 days from device insertion on CD's 3. 7, or 13, and in untreated controls Average difference between class 3 follicles at day of CIOR-8 insertion and during the CIOR-B treatment in cows with or without a corpus luteum (CL) Average size of the largest follicle at CIOR-8 removal in cows with or without a corpus luteum (CL) Plasma progesterone (ng/ml) during the 1 0 days of CIDR-8 treatment in cows with or without a corpus luteum (CL) The average number of class 1 ( < 6 mm) for cows at different days of the periovulatory period Table 1.8 Interval to oestrus after CIOR-8 removal in cows Table 1.9 Average number of class 1 ( < 6 mm), class 2 (6-9 mm) and class 3 ( > 9 mm) follicles at CIOR-8 removal in cows with short ( 1-3 days) or long (4-9 days) post-removal intervals to oestrus Table 1.10 Average number of ovarian follicles in classes 1, 2, and 3 during the CIOR-8 treatment for ovulatory and nonovulatory ovary for cows with short ( 1-3) or longb (4-9) post-treatment intervals to oestrus after CIOR-8 removal

25 Size of largest follicle in cows with differing post- treatment intervals from CIDR-8 removal to oestrus xxii Table Table Table Average number of follicles waves leading to the ovulatory follicle during CIDR-8 treatment for cows with different intervals to oestrus after CIDR-8 removal Average plasma progesterone concentrations (PPC) on selected days during one oestrous cycle in control cows Table Average plsama progesterone concentrations (PPC[mean (SEM); ng/ml]) at stated time of CIDR-8 insertion in different treatment groups and on selected days (CD) during one oestrous cycle in normally cycling (NC) group Table Average plasma progesterone concentrations (PPC[Mean (SEM); ng/ml]) at the time of CIDR-8 removal (TD 10) in ovariectomized (OVX) and cycling cows Table 2. 1 Table 2. 2 Table 2.3 Table 2. 4 Table 2.5 Table 2. 6 Table 2. 7 Table 2. 8 Distribution of intervals to oestrus and insemination between treatments within trials and between groups of heifers within a treatment Tailpaint and raddle scores at the time of initial oestrus detection among heifers included in Trial Average post-treatment interval to onset of oestrus in groups of heifers which received three different treatments to synchronize oestrus (Trial 1) Average duration of behavioural oestrus in each group of synchronized and control heifers 11 6 Tailpaint and raddle scores at the time of initial oestrus detection among cows in Trial Average post-treatment interval to onset of oestrus in groups of cows which received three different treatments to synchronize oestrus (Trial 2) Average duration of behavioural oestrus in each group of synchronized and control cows Comparison between plasma progesterone concentrations (PPC, Mean (SEM); ng/ml) by radioimmunoassay and tail paint and raddle scoring at

26 xxiii the time of detection of oestrus in cycling heifers Table 3. 1 Table 3. 2 Table 3.3 Table 3. 4 Table 3.5 Table 3. 6 Table 3. 7 Table 3. 8 Conception rates (%), services per conception and least squares means (SEM) of planned start of mating (PSM) to first insemination (AI), PSM to conception and interval from 1st AI to conception (days) in treatment and control groups in Farm A (Year-round herd) Conception rates (%), services per conception and least squares means (SEM) of planned start of mating (PSM) to first insemination (AI), PSM to conception and interval from 1st AI to conception (days) in treatment and control groups in Farm B (Year-round herd) Conception rates (%), services per conception and least squares means (SEM) of planned start of mating (PSM) to first insemination (AI), PSM to conception and intervals from 1st AI to conception (days) in treatment and control groups in Farm C (Year-round herd) Conception rates (%) and least squares means (SEM) of planned start of mating (PSM) to first insemination (AI) and PSM to conception (days) in treatment and control groups in Farm D (Seasonal-herd, Autumncalving) Conception rates (%) and least squares means (SEM) of planned start of mating (PSM) to first insemination (AI) and PSM to conception (days) in treatment and control groups in Farm D (Seasonal-herd, Springcalving) Conception rates (%), services per conception and least squares means (SEM) of planned start of mating (PSM) to first insemination (AI), PSM to conception and intervals from 1st AI to conception (days) in treatment and control groups in Year-round herds Conception rates (%), services per conception and least squares means SEM) of planned start of mating (PSM) to first insemination (AI), PSM to conception and intervals from 1st AI to conception (days) in non-cycling cows in Year-round herds Conception rates (%), services per conception and least squares means (SEM) of planned start of

27 xxiv mating (PSM) to first insemination (AI), PSM to conception and intervals from 1st AI to conception (days) in non-cycling cows in Seasonal herd Table 3.9 Table Table Table Table Table 4.1 Table 4. 2 Table 4.3 Table 4.4 Table 4.5 Table 4.6 Percentage (%) of cows in oestrus and inseminated in treatment group during the 5-Day period and in control cows during the 38-Day period Percentage (%) of cows in oestrus and inseminated in treatment group during the first period of artificial insemination (3-Day period) Effect of dose rates of prostaglandin (PGF) at CIDR-8 removal on oestrus and fertility in cows Effect of treatment with progesterone on conception rate at second artificial insemination (All in treatment and control groups Least squares means (SEM) of plasma progesterone concentrations (ng/ml) in non-pregnant (NP) and pregnant (P) cows treated with CIDR-8, and prostaglandin (PGF) or pregnant mares serum gonadotrophin (PMSG) Percentage (%) of cows inseminated over the second period of artificial insemination after being identified as non-pregnant by 8.E.S.T Test Kit Means (SEM) of milk progesterone concentrations (ng/ml) by radioimmunoassay (RIAl of non-pregnant and pregnant cows at one day after CIDR-8 removal Sensitivity and specificity for different concentrations of progesterone in milk (P4; ng/ml) determined by radioimmunoassay Comparison between milk progesterone concentrations (ng/ml) by radioimmunoassay (RIA) one day after CIDR-8 removal and pregnancy status at rectal palpation 42 days after artificial insemination Percentage (%) of non-pregnant (NP) and pregnant (P) animals at 3 concentrations of progesterone determined by radioimmunoassay Progesterone classification into low and high level by using Best test and radioimmunoassay (RIA)

28 XXV Table 4. 7 Table 4. 8 Table 4. 9 Table 5. 1 Table 5. 2 Table 5.3 Table 5.4 Table 5.5 Table 5. 6 Distribution of samples by concentrations of progesterone (P4) in milk as determined by RIA (nonpregnant and pregnant), after being classified by BEST test Sensitivity and specificity for different concentrations of progesterone in plasma (P4; ng/ml) determined by radioimmunoassay Means (SEM) of plasma progesterone concentrations (ng/ml) by radioimmunoassay (RIA) of non-pregnant and pregnant cows at one day after CIDR-8 removal Average number of ovarian follicles in classes 1, 2 and 3 and total number of follicle at initial examination in treated cows and at the same time for the contemporary control cows in the early and late post-partum (PP) period in anoestrous cows Average number of ovarian follicles per animal in classes 1, 2 and 3 and total number of follicles during CIDR-8 insertion in treated cows and in the same period for contemporary control cows in early post-partum period (PP) in anoestrous cows Average number of ovarian follicles in classes 1, 2 and 3 and total number of follicles during the period of CIDR-8 insertion in treated cows and in the same period for contemporary control cows in late postpartum period (PP) in anoestrous cows Average number of ovarian follicles per animal in classes 1, 2 and 3 and total number of follicles during CIDR-8 insertion in treated cows and in the same period for contemporary control cows in the early and late post-partum (PP) period in anoestrous cows Average number of class 1 ( < 6 mm), class 2 (6 to 9 mm), and class 3 ( > 9 mm) of ovarian follicles and the average size of the largest normal and luteinized class 3 follicle at CIDR-8 removal and one day later in anoestrous treated cows in the early post-partum period (Trial 1) Average number of class 1 ( < 6 mm), class 2 (6 to 9 mm), and class 3 ( > 9 mm) of ovarian follicles and the average size of the largest normal and luteinized class 3 follicle at CIDR-8 removal and one day later in anoestrous treated cows displaying ( +) or not (-)

29 xxvi oestrus after CIDR/PMSG treatment in the early postpartum period (Trial 1) Table 5.7 Table 5.8 Table 5.9 Table 5.10 Average number of ovarian follicles in class 1 ( < 6 mm), class 2 (6 to 9 mm), and class 3 ( > 9 mm), and the average size of the largest normal and luteinized class 3 follicle at and 1 day after CIDR-8 removal in anoestrous treated cows which subsequent formed ( + ) or did not form (-) a corpus luteum (CL) after CIDR/PMSG treatment in the early post-partum period (Trial 1) Average of class 1 ( < 6 mm), class 2 (6 to 9 mm), and class 3 ( > 9 mm) of ovarian follicles and the average size of the largest normal and luteinized class 3 follicle at CIDR-8 removal and 1 day later in anoestrous treated cows in the late post-partum period (Trial 2) Average number of class 1 ( < 6 mm), class 2 (6 to 9 mm), and class 3 ( > 9 mm), and the average size of the largest normal and luteinized class 3 follicle at CIDR-8 removal and 1 day later in anoestrous treated cows which displayed oestrus and formed a corpus luteum ( +; CL) or did not display oestrus and form a CL after Cl DR/PMSG treatment in the late postpartum period (Trial 2) Average plasma progesterone concentrations (PPC; ng/ml) at the time if CIDR-8 insertion and removal in treated and in the same period for contemporary control in anoestrous cows in the early and late postpartum period (PP)

30 1 GENERAL INTRODUCTION The present thesis was undertaken as a series of five projects in which selected studies were undertaken on the use of the CIDR intravaginal device in dairy cattle. The first study was designed to characterize the interactions involving the dynamic changes of follicles in the ovaries of cycling cows, measured by ultrasound during and after CIDR insertion and until oestrus, and the associated changes in plasma progesterone concentrations at the different stages of the oestrous cycle in entire cows, and in ovariectomized animals. The second study concerns an evaluation of the effectiveness of tailpainting combined with the use of an aerosol raddle technique in detecting oestrus in synchronized animals. Other aims were to compare the patterns of onset of oestrus with different synchrony treatment, duration of oestrus, and some observations of oestrous behaviour in animals forming sexually active groups. The third study evaluated a reproductive programme in dairy herds with a daily quota, and its effects on oestrus detection rates and return to service intervals, oestrous responses to two dose rates of prostaglandin combined with CIDR, and the effect of progesterone supplementation on fertility. The fourth study was designed to evaluate the effectiveness of using milk progesterone tests to identify non-pregnant animals, and consequently improve oestrus detection rates in cows previously synchronized in a controlled breeding management system involving the use of CIDR devices for controlling return to service intervals. The fifth study concerns a study of dynamic changes of follicles in the ovaries by using ultrasonography during two different phases of the post-partum period (early and late), an examination of the effects of CIDR insertion and injection of PMSG, and measurement of post-treatment response rates in oestrus and ovulation in anoestrous dairy cows.

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