Animal Breeding Reproduction Presentation.pptx

animal reproduction

1. PRINCIPLES AND PRACTICES OF ANIMAL BREEDING

2. Course outline

3. Nucleus Breeding system CLOSED NUCLEUS OPEN NUCLEUS

4. MALE REPRODUCTIVE SYSTEM

5. MALE REPRODUCTIVE SYSTEM Penis: Copulation Glans Penis: Sensation Cowper's gland: lubrication; pH Prostate gland: Lubrication; Nutrition Sigmoid flexure: extrusion/retraction Testes : Sperm, testosterone Scrotum : Protection, Temperature regulation Vas deference: transportation Seminal Vesicle: Nutrition to sperm Prepuce: Protection of glans penis

6. testis Spermatic chord Seminiferous tubules Nerves Vas deference Blood Vessel Testis Epididymis Sertoli cells Leydig cells Rete Testis Spermatozoa Lumen Epididymis: Sperm maturation, transportation Rete Testis : Sperm transportation Seminiferous tubules: Sperm production Sertoli cells: Sperm production Leydig cells: Testosterone production

7. Comparative study Stallion Bull Ram Boar

8. Female reproductive system

9. Birth canal Ileum Ischium Pubis Sacrosciatic Ligament Sacrum

10. Female reproductive system Reproductive organs ovaries Reproductive tract Fallopian tube Uterus Cervix vagina External genitalia Vulva Clitoris Labia

11. Ovary Follicle Oocyte, estrogen Corpus Luteum Progesterone

12. Ovary: comparison Cow Sow Mare

13. Oviduct Uterine Horn Utero-Tubal Junction Ampullary-Isthmic Junction Isthmus Ampulla Infundibulum Fimbria Follicle Ovary

14. Uterus two uterine horns a body Transportation, absorption and phagocytosis of sperm Reception of fertilized ovum Nutrition and protection of fetus Placental attachment Expulsion of fetus

15. Uterus: Comparison Bicornuate 1 body, 2 horns Cattle Sheep Swine Longer horns Human Rabbit Duplex No body, 2 horns Simplex 1 body, No horns

16. cervix Internal Cervical Os Annular rings External Cervical Os Close uterine lumen against infection Passage for sperm at estrus Cervical plug at pregnancy Dilatation at parturition

17. Cervix : comparison No obstacles Longitudinal Folds Interdigitating pads Fornix Vagina No Fornix Vagina Cow Mare Sow Cow has 4-5 annular rings

18. Vagina Fornix Vulva-Vaginal Sphincter Anterior Vagina Posterior Vagina (Vestibule) Copulatory organ Passage for fetus at parturition Capable of great dilatation Lubrication Pheromone

19. Vulva Anus Labia Clitoris Labia majora (Outer lips) Labia minora (Inner lips) Vestibule Clitoris: Female Sensory organs Indication of estrum

20. Vulva: Comparison Mare Cow Sow Ewe

21. ENDOCRINOLOGY OF REPRODUCTION

22. Endocrinology of reproduction Exocrine glands: Glands with duct Local action Secrete enzymes Endocrine glands: Ductless glands Via blood stream Secrete hormones At first know the glands

23. Endocrinology of reproduction Hormones: produced in specialized tissue; secreted into blood; specific biological action in target organs Then hormones

24. Action of hormones: Distance covered Autocrine Endocrine Paracrine Unattached receptors Attached receptors

25. Chemical classification of hormones Lipophobic Amines: Tyrosine or Tryptophan derived: e.g. Melatonin Peptide: Short chain of linked amino acid E.g. Oxytocin Protein: Poly peptide or glycoprotein: e.g. FSH, LH Lipophillic Steroid: Cholesterol derived: eg. Testosterone, estrogen, progesterone Lipid: Fatty acid derived: e.g. Prostaglandin

26. Mechanism of hormone action Lipophilic Lipophobic

27. Hypothalamus Secreted hormones Effect Prolactin-releasing hormone (PRH) Stimulate prolactin release Prolactin-inhibiting hormone (PIH) Inhibit prolactin release Gonadotropin-releasing hormone (GnRH) Stimulate  FSH release Stimulate LH release Oxytocin Uterine contraction Lactation (letdown reflex) Melatonin day length gonadotropic activity in mare and sheep

28. Pituitary gland Anterior Pitutary (Adenohypophysis): Somatotrophs: GH, ACTH Mammotrophs : Prolactin Thyrotrophs : TSH Gonadotrophs: FSH & LH Posterior piturary : No secretion, only acts for storage and release of hormones: Oxytocin and melatonin

29. Pituitary gland action Hypothalamus Anterior pituitary Posterior pituitary Portal vein

30. Pituitary hormones FSH: In Female: Ovarian and follicular growth and development. Secretion of estrogen from ovary in presence of LH In Male: Spermatogenesis upto secondary spermatocyte stage Luteinizing Hormone (LH): In female: Tonic or basal level: with FSH induce estrogen secretion from follicle Preovulatory surge: ovulation and luteinization of follicular cells In male: Stimulates growth of the interstitial (Leydig) cells and the secretion of testosterone.

31. Gonadal hormones Estrogen Maturation, growth and development of reproductive organs Stimulation of reproductive physiology: Growth of uterine muscle Development of endometrial lining of tubes Increase vas cularity Behavioral estrus Dilatation of cervix Contraction of uterus along with oxytocin Secondary sex characters Progesterone Secretion from endometrial glands (Uterine milk) Maintenance of pregnancy Inhibit smooth muscle activity Needed to prime estrogen action Inhibit FSH and LH secretion: negative feedback

32. Placental hormones Equine Chorionic Gonadotropin ( eCG ): Secreted from endometrial cups in mares from 40-140 days of gestation with maximum level at 80 th day of pregnancy Have like both LH and FSH like activity but FSH more potent In mares it results folliculogenesis and ovulation and hence multiple CL It is used to treat inactive ovaries as well as used for superovulation in donor animals in embryo transfer. Prostaglandins: Potent luteolytic agent resulting in a decrease in progesterone levels

33. Regulation of hormones . Endocrine feedback mechanism: Increase or decrease concentration of one hormone either stimulates or subsides release of another hormone. Neuroendocrine reflex: The nervous system controls the release of hormone through neural pathways eg. Oxytocin in milk letdown and LH release following copulation Immunoendocrine Control: The endocrine and immune system interact extensively to regulate each other. Several endocrine organs are involved in some aspect of this regulatory process e.g. hypothalamus, pituitary, gonads etc.

34. Feedback Mechanism

35. Growth stages Infantile stage Post pubertal stage puberty Pre pubertal stage maturity No hormonal change No response to hormone High hormonal change No response to hormone Gametogenesis start as response to hormonal change Gametogenesis increase in quality and quantity Mature type hormonal change Mature stage of gametogenesis

36. Estrus cycle

37. Ovarian changes in estrus cycle

38. Phases of estrus cycle

39. Stages of Estrus cycle . Stage Ovary Uterus Cervix Vulva Proestrum Developing follicle Regressing CL Increasing tonicity Open Pink Slight swelling Slight wet Estrum Graffian follicle Ovulaton at end Tonic Open Red Swollen Mucus secretion Metestrum Developing CL Lose tonicity Closed Red patches Shrinking Drying Diestrum Developed CL Flaccid Closed Pale Shrinked dry

40. Folliculogenesis

41. folliculogenesis

42. folliculogenesis

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45. Signs of estrus Sniffing vulva of other females Bellowing and restlessness Swollen vulva Mounting or let be mounted Resting chin on other animals Soliciting Mounting head to head Dirty flank Mucus from vulva Licking

46. Detecting estrus Signs of estrus Tail painting and scratch cards Rectal palpation Body temperature Hormone Assay Ultrasonography Pedometer Heat detector

47. Estrus synchronization ↓ generation/calving interval Shorten breeding season ↓ estrus detection error Common management of calves Embryo transfer ↓ Labor cost ↑ Conception rate ↑ calves/cow

48. Hormones used for E.S. Prostaglandin F2 α Progesterone Gonadotropin Releasing Hormone (GnRH) Estrogen Luteolysis of CL ˃ 4-6 days Heat after 2-5days of use Progesterone block Heat after removal LH and FSH for follicle and ovulation Generally used in combination Luteolysis of CL ˂ 6 days Limited use & in combination Rarely used

49. Hormones used for E.S. Prostaglandin F2 α Progesterone Gonadotropin Releasing Hormone (GnRH) Estrogen Luteolysis of CL ˃ 4-6 days Heat after 2-5days of use Progesterone block Heat after removal LH and FSH for follicle and ovulation Generally used in combination Luteolysis of CL ˂ 6 days Limited use & in combination

50. Hormones used for E.S. Prostaglandin F2 α Progesterone Gonadotropin Releasing Hormone (GnRH) Estrogen Luteolysis of CL ˃ 4-6 days Heat after 2-5days of use Progesterone block Heat after removal LH and FSH for follicle and ovulation Generally used in combination Luteolysis of CL ˂ 6 days Limited use & in combination Rarely used

51. EC Protocols: PGF2 α One shot PGF2 α Two shot PGF2 α

52. EC Protocols: PGF2 α + GnRH Select sync Ovsync Co-sync

53. EC Protocols: PGF2 α + Progesterone 7-day CIDR +PGF2 α 6-day CIDR +PGF2 α CIDR: Controlled intrauterine drug release

54. EC Protocols: PGF2 α + Progesterone . 14-day CIDR + PGF2 α

55. EC Protocols: PGF2 α + GnRH + Progesterone . Ovsync + CIDR Select sync + CIDR

56. EC Protocols: PGF2 α + GnRH + Progesterone 5-day Co-sync + CIDR Co-sync + CIDR 14-day CIDR + PGF2 α + GnRH

57. EC Protocols: PGF2 α + GnRH + Progesterone . 14-day MGA + PGF2 α 14-day MGA + PGF2 α + GnRH MGA: Melengestrol Acetate

58. EC Protocols: PGF2 α + GnRH + Estrogen Heat sync ECP: Estradiol cypionate

59. Course outline

60. ARTIFICIAL INSEMINATION (AI) Lazanno Spalbanzani First AI in 1780 AD In dog I have never met your father

61. Artificial insemination (AI) Artificial introduc tion of artificially collected semen to female reproductive tract Selection of bull Semen collection Semen analysis Sperm separation Semen processing Semen analysis Semen Storage and transportation Thawing semen Insemination Steps of AI

62. Advantages of AI Genetically superior sire Genetic improvement Sexed se men Control of STD Superior selection of bull ↓ number of bulls needed Remote insemination ↓ cost Semen of injured, old, heavy sire ↑ efficiency of synchronization ↑ progeny testin g of sire Insemi nate dam refuse to mate Safe mating ↓ Inbreeding Accurate breeding program Insemination of small flock

63. disadvantages of AI Neet of technology Technical skill Remote disease introduction Lesser conception rate Infection due to instruments Loss of indigenous AnGR Parturition difficulties with larger bulls Reduced bull market

64. Semen collection Collection from vagina False mount method Teaser cow Dummy cow Electro ejaculator Digital manipulation Especially in birds Dummy cow Teaser cow Electro ejaculator

65. Semen collection Water inlet Warm water Chamber Rubber cone Collection Tube Rubber Liner Outer casing Artificial Vagina (AV) Semen collection cups

66. Semen collection Courtship with teaser cow False mounting for arousal Erect penis means arousal Collector ready with AV Divert penis to AV

67. Semen collection Electric Probe Method Digital manipulation

68. Semen Analysis Ejaculate volume: 5 ml (1-15 ml) Uniform and opaque appearance Milky to creamy color (red: blood) No contamination >500 million sperm per ml. (300-2500 million/ml) >70% live sperm >50% of motile sperm with progressive motility >70% Normal morphology Major abnormality <20% Minor abnormality<30%

69. Abnormal sperm

70. Abnormal sperm

71. Abnormal sperm

72. Abnormal sperm

73. Abnormal sperm

74. Abnormal sperm

75. Sperm separation Swim up Density gradient centrifugation Density gradient- Swim up Glass wool filtration Microfluidics Magnetic activated cell sorting Florescent activated cell sorting Zeta Potential selection

76. Semen dilution/extension Dilution of bull semen 1:15-1:50; ram and buck 1:3-1:5 2-5 million sperm/ml for bull in straw (0.5 ml straw) Single ejaculate: hundreds of straw Extender: Nutrients: Energy source: monosaccharides, amino acids, glycerol Osmotic pressure and buffers: Sodium citrate, sodium phosphate Cold shock prevention: Milk, skim milk, egg yolk Inhibit bacterial growth: antibiotics Increase volume and dissolve: distilled water Cryoprotectant: glycerol

77. Semen preservation and storage .

78. Insemination of dam: armory

79. Insemination of dam

80. Insemination of dam

81. Transportation of sperm

82. Fertilization

83. Pre-attachment development of embryo (day 7-8) (day 6-7) Collection of embryo for transfer Day 7

84. Embryo transfer technology (ETT) Walter Heape First ET in 1890 AD In rabbit You are my child, but I don’t know your father and mother

85. embryo transfer: pros and cons Increase genetic potential of herd Embryo from injured or old dam Preservation of frozen embryo Multiple offspring from single dam Short time for genetic progress Higher level of management High technology required Disease transfer Potential doner may not respond Distant shipping of frozen embryo

86. Steps of ett

87. Select donor and recipient dams Genetic contribution Disease free High genetic potential for required trait Good reproductive capacity No genetic contribution Disease free Good bearing capacity Enough production for offspring Donor Recipient Synchronize them GnRH Progesterone PGF2 α Estrogen

88. Superovulation & a.i. Multiple tapering dose of FSH Multiple ovulation Insemination of donor Confirmation by Rectal palpation or USG

89. Flushing: collecting embryo Foley catheter Embryo filter Lignocaine To reduce pain Penecillin Prevent infection Kill remaining embryo 7-10 embryos

90. Processing and storage of embryo Transferring embryo solution to search plate Rinsing embryo filter with washing medium Searching for embryo Rinsing embryo filter with washing medium

91. Embryo quality Embryo size ≈ 0.2 mm Good quality embryo Spherical Symmetrical Uniform sized cells Embryo holding media Dulbecco’s phosphate buffer saline (DPBS) + Glucose + Sodium p yruvate + Penicillin + Streptomycin + Bismuth Sulphate Agar (BSA)

92. Transferring embryo to recipient dam Surgical Non-surgical

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105. . HDAI 5 10 8 PGF2 α PGF2 α TAI GnRH GnRH Days CIDR ® PGF2 α 8 ±2 hrs 66 ±2 hrs GnRH Progesterone PGF2 α Estrogen

106. . 14 28 PGF2 α Days 5 16 11 PGF2 α PGF2 α HDAI Days HDAI PGF2 α 35 36 38 GnRH PGF2 α ECP TAI Estradiol cypionate

107. . HDAI 14 33 39 PGF2 α PGF2 α HDAI CIDR ® GnRH Days

108. . HDAI 14 33 39 PGF2 α PGF2 α Days 39 MGA® Melengestrol acetate

109. . HDAI 14 33 39 PGF2 α PGF2 α Days 36

110. . Preovulatory LH surge ↑ PGE 2 ↑ Blood flow to ovary and dominant follicle Edema ↑ Follicular Pressure Follicular wall weakness ↑ PGF 2 α Shift from E 2 to P 4 by dominant follicle ↑ P 4 ↑ Collagenase ↑ Contraction of Ovarian smooth muscle Release of lysosomal enzyme OVULATION

111. . ↓ P 4 from CL ↑ GnRH ↑ FSH and LH Proestrual Follicular development ↑ E 2 to threshold ↓ FSH Theca externa Theca interna Granulosa cells Cumulus oophorus Antrum Corona radiata Ovum ↑ Inhibin Preovulatory LH surge