Introduction1
Chapter 1　Estrus and Ovulation Control3
　1-1　Principle of estrus diagnosis 4
　　1-1-1　General diagnosis technology5
　　1-1-2　Modern estrus monitoring technology9
　1-2　Estrus control technology14
　　1-2-1　Estrus induction technology14
　　1-2-2　Simultaneous estrus technology16
　1-3　Ovulation control technology20
　　1-3-1　Ovulation timing control and Ovsynch21
　　1-3-2　Ovulation count control 22
Chapter 2　Fertilization Technology25
　2-1　Artificial insemination 25
　　2-1-1　Advantages and disadvantages of artificial insemination 26
　　2-1-2　Brief introduction of development history for artificial insemination
technology in livestock and poultry27
　　2-1-3　Technical process of artificial insemination28
　2-2　In vitro fertilization technique52
　　2-2-1　A brief history of in vitro fertilization52
　　2-2-2　In vitro maturation of oocytes54
　　2-2-3　In vitro capacitation of sperm56
　　2-2-4　Procedure of in vitro fertilization 58
　　2-2-5　Main factors affecting in vitro fertilization 60
　　2-2-6　Application, problems, and prospects of in vitro fertilization63
　2-3　Microinsemination 64
　　2-3-1　Operating procedure65
　　2-3-2　Significance of MI67

Chapter 3　Pregnancy Diagnosis69
　3-1　Basis for pregnancy diagnosis69
　3-2　Principles of pregnancy diagnosis70
　3-3　Methods of pregnancy diagnosis70
　　3-3-1　Routine techniques for pregnancy diagnosis 71
　　3-3-2　Smart pregnancy diagnosis techniques98
Chapter 4　Parturition Control102
　4-1　Principle of parturition control102
　4-2　Application of parturition control103
　4-3　Significance of parturition control104
　4-4　Methods of parturition control104
　　4-4-1　Treatment scheme of prostaglandin105
　　4-4-2　Treatment scheme of oxytocin106
　　4-4-3　Treatment scheme of corticosteroid106
Chapter 5　Oocyte and Embryo Cryopreservation108
　5-1　History for embryo and oocyte cryopreservation108
　　5-1-1　Oocyte cryopreservation108
　　5-1-2　Embryo cryopreservation109
　5-2　Principle and technology of cryopreservation for ova and embryos110
　　5-2-1　Technical terms 110
　　5-2-2　Changes during freezing and cryoinjuries 110
　　5-2-3　Cryoprotectants112
　　5-2-4　Principles of cryopreservation112
　5-3　Procedures for embryo cryopreservation113
　　5-3-1　Conventional freezing113
　　5-3-2　Ultrarapid freezing115
　　5-3-3　Vitrification cryopreservation procedure116
　5-4　Procedures for oocyte cryopreservation118
　　5-4-1　Slow freezing118
　　5-4-2　Vitrification cryopreservation118
　5-5　Prospects and conclusions of oocyte cryopreservation123
Chapter 6　Sex Control in Mammals124
　6-1　Brief history of studies on sex determination125
　6-2　Sex control techniques in mammals125
　　6-2-1　Sex control before fertilization—the separation of X sperm and
Y sperm 126
　　6-2-2　Internal female sexual manipulation of the female reproductive tract130
　　6-2-3　Mating with semen by RNAi injection into testis131
　　6-2-4　Sex identification of preimplanted embryo131
　　6-2-5　Sex control by ultrasonography identification135
　　6-2-6　Other biotechniques135
　6-3　Application prospect of sex control techniques135
Chapter 7　Embryo Transfer137
　7-1　Research history of embryo transfer138
　　7-1-1　Development of fresh embryo and embryo transfer138
　　7-1-2　Development of frozen embryo transfer138
　　7-1-3　Development of combination of micromanipulation and
embryo transfer 138
　　7-1-4　Embryo transfer organizations contributed to the
technique development139
　7-2　Rationale of embryo transfer139
　　7-2-1　Gestational development of reproductive organs after estrus 140
　　7-2-2　Preimplantation status of an early embryo 140
　　7-2-3　Immune tolerance of the maternal uterus to the embryo140
　　7-2-4　Pregnancy signals are emitted by the embryo140
　　7-2-5　Stability of embryonic genetic material 141
　7-3　Principles of embryo transfer 141
　　7-3-1　Physiological basis of embryo transfer141
　　7-3-2　Time frame for embryo transfer142
　　7-3-3　Strict quality control of the embryo transfer process142
　7-4　Factors affecting the success of embryo transfer142
　　7-4-1　Maternal hormonal environment required for embryo implantation143
　　7-4-2　Embryonic preparation for implantation necessitates
“blastocyst activation”143
　　7-4-3　Uterine receptivity144
　　7-4-4　Cellcell interactions144
　7-5　Procedure of embryo transfer in farm animals145
　　7-5-1　Selection of donor145
　　7-5-2　Selection of recipient145
　　7-5-3　Superovulation of donor 146
　　7-5-4　Artificial insemination of donor147
　　7-5-5　Synchronization of recipient147
　　7-5-6　Recovery of embryos149
　　7-5-7　Embryo quality control151
　　7-5-8　Embryo transfer to recipient153
　　7-5-9　Postoperative care of donor and recipient154
　7-6　Main problems of embryo transfer155
　　7-6-1　Less consistent superrowing results155
　　7-6-2　Low embryo recovery rate155
　　7-6-3　Donor reuse problems155
　　7-6-4　Supporting technology problems155
　　7-6-5　Lack of technical personnel and inadequate promotion system155
Chapter 8　Stem Cell157
　8-1　Research history of stem cells157
　8-2　Morphological and functional characteristics of stem cells160
　　8-2-1　Morphological and functional characteristics of mesenchymal
stem cells160
　　8-2-2　Morphological and functional characteristics of embryonic stem cells162
　　8-2-3　Morphological and functional characteristics of iPS cells164
　8-3　Technical points of stem cell isolation and culture170
　　8-3-1　Main points of culture techniques for mesenchymal stem cells170
　　8-3-2　Main points of culture techniques for embryonic stem cells171
　　8-3-3　Technical points of iPS cell culture172
　8-4　Main problems of stem cell technology181
　　8-4-1　Main problems of mesenchymal stem cell technology181
　　8-4-2　Main problems of ES cell technology182
　　8-4-3　Main problems existing in iPS cell technology182
　8-5　Application prospects of stem cell technology183
　　8-5-1　Application prospects of mesenchymal stem cells technology183
　　8-5-2　Application prospects of embryonic stem cell technology184
　　8-5-3　Application prospects of iPS cell technology186
Chapter 9　Animal Cloning189
　9-1　Parthenogenesis189
　　9-1-1　PA of ZPfree oocytes in pigs190
　　9-1-2　PA of ZPintact oocytes in pigs192
　9-2　Embryos splitting192
　　9-2-1　Basic procedure to split embryos193
　　9-2-2　Significance and application of embryos splitting193
　9-3　Nuclear transfer194
　　9-3-1　Brief introduction of nuclear transfer194
　　9-3-2　Somatic cell nuclear transfer195
　9-4　Main problems of animal cloning199
　9-5　Significance and application of animal cloning200
Chapter 10　Transgenic Animal202
　10-1　Point and application of transgenic animal technology202
　　10-1-1　Application of transgenic animal technology in livestock204
　　10-1-2　Application of transgenic animal technology in biology207
　　10-1-3　Application of transgenic animal technology in pharmaceuticals208
　10-2　Main technical links of transgenic animal technology210
　　10-2-1　Targeting gene selection 211
　　10-2-2　Gene modification and expression vector construction211
　　10-2-3　Introduction of exogenous genes212
　　10-2-4　Gene integration and expression analysis215
　　10-2-5　New progress in the research of animal transgenic technology216
　10-3　Main questions and developmental trend in transgenic animal technology221
　　10-3-1　Main question in transgenic animal technology221
　　10-3-2　Developmental trend in transgenic animal technology222
Chapter 11　Hormone Immunization and Genetic Immunization224
　11-1　Hormone immunization224
　　11-1-1　Steroid hormone immunization225
　　11-1-2　Inhibin immunization226
　　11-1-3　Gonadotropin inhibiting hormone immunization 228
　　11-1-4　AntiMüllerian hormone immunization 228
　　11-1-5　Immunocastration228
　11-2　Genetic immunization 230
　　11-2-1　Principle of genetic immunization 231
　　11-2-2　Methods of genetic immunization231
　　11-2-3　Advantages and safety of genetic immunization 232
　　11-2-4　Genetic immunization for regulating animal reproduction233
Chapter 12　Avian Reproductive Biotechnology238
　12-1　Techniques to control avian seasonal reproduction239
　　12-1-1　Mechanism of avian reproductive seasonality239
　　12-1-2　Procedures to control avian seasonal reproduction240
　12-2　Avian embryo manipulation technique240
　　12-2-1　Artificial insemination and embryo transplantation241
　　12-2-2　Culture of avian embryos in the early stage241
　12-3　Avian chimeric technology243
　　12-3-1　Classification of avian chimeras 243
　　12-3-2　Production of avian chimeras247
　　12-3-3　Identification of chimeras in poultry248
　12-4　Sex control technology of poultry249
　　12-4-1　Sex reversal249
　　12-4-2　Sex identification 250
　　12-4-3　Problems of sex control253
　12-5　Transgenic technology of avian254
　　12-5-1　Method of gamete vector 254
　　12-5-2　Technique of microinjection for fertilized egg256
　　12-5-3　Transfection of embryonic placental cells using retroviral vectors257
　　12-5-4　Transfection of blastoderm cells with lipofectin packaging vectors 258
　　12-5-5　Method of blastoderm cell and primordial germ cell vectors259
　　12-5-6　Problems and prospects 260
Abbreviations262
References269
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