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Artificial Insemination

Artificial Insemination. John Parrish. Objectives of Artificial Insemination. Genetic improvement of livestock Disease control mechanism Possible to increase fertility Decrease breeding expense. Current Status of US Industry. Dairy Cattle 7 million (50%) - in Denmark, Japan, 90-100%

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Artificial Insemination

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  1. Artificial Insemination John Parrish

  2. Objectives of Artificial Insemination • Genetic improvement of livestock • Disease control mechanism • Possible to increase fertility • Decrease breeding expense

  3. Current Status of US Industry • Dairy Cattle • 7 million (50%) - in Denmark, Japan, 90-100% • Beef Cattle • 1.3 million (3%) • Swine • 60 - 80% • Turkey • 100% • Horses • Increasing rapidly

  4. Advantages of AI • Genetic Improvement • Wide spread use and availability of genetically superior sires • 1 bull can breed 500,000 cows in a lifetime • After death, semen can be used • Oldest frozen semen 40 - 45 years old • Rapid proof of sire • Progeny testing examines offspring for desired traits • With natural mating would only have 100’s of offspring

  5. Advantages of AI (cont.) • Availability of sires • Sires anywhere in world • Danger of bull (male) removed • Disease reduction • Crossbreeding • Can try without buying sire • Improved management • Start to keep records

  6. Advantages of AI (cont.) • Economics • Cost of very good sire is reduced because extend semen • Cost to maintain sire’s reduced as don’t need as many to breed all the females

  7. Disadvantages • Estrus detection must be good • Trained inseminator • Bull semen the best, other species not as good • Use of poor male may increase if not tested well • Technology to store cooled or frozen semen • Difficult to maintain

  8. Age When Semen Can Be Collected Bull 12 months Boar 6 - 8 months Ram 6 - 9 months Stallion 20 - 24 months Dog 8 - 12 months

  9. Effect of Age on Sperm Output Bull Sperm Output 0 10 20 30 40 50 60 Age in Months

  10. Semen Collection • Sexual arousal • Sight, sound, smell, touch • Best mount - Live

  11. Collection on a Live Mount

  12. Collection on a Live Mount

  13. Semen Collection • Sexual arousal • Sight, sound, smell, touch • Best mount - Live • Alternative mount - dummies • Bull • Stallion • Boar • Ram

  14. Collection of a Bull on a Dummy

  15. Stallion Collection on a Phantom

  16. Boar Collection on a Dummy

  17. Ram Collection on a Dummy

  18. Semen Collection (cont.) • Sexual Preparation • False mounts • In bulls can increase sperm collected by 100% • Novelty • Change mounts, location, other males present • Breed and species differences • Beef bulls less libido than dairy bulls • Arab stallions more libido than Quarter horse stallions • Rams less libido than bulls or male goats

  19. Semen Collection (cont.) • Frequency of collection • As increase frequency/wk • decrease sperm/ejaculate • Increase sperm recovered/wk

  20. Effect of Collection Frequency on Sperm Output Item Dairy Beef Sheep Swine Horses # of collections 1-6 1-6 7-25 2-5 2-6 Volume (ml) 5-8 3-6 0.8-1.2 150-300 30-100 Concentration (million/ml) 1000-2000 800-1500 2000-3000 200-300 200-400 Total sperm/ejac. (billion) 7-15 5-10 1.6-3.6 30-60 5-10 Total sperm/wk (billion) 15-40 10-30 25-40 100-150 15-30 Motile sperm(%) 50-75 40-75 60-80 50-80 40-75 Normal sperm(%) 70-95 65-90 80-95 70-90 70-90

  21. Artificial Vaginas Water Water Inner Liner Collection cone Collection vial

  22. Characteristics of AV • Temperature • Bull, stallion, ram - 45°C • Pressure • Friction • Lubrication essential

  23. Electroejaculation • Useful if male won’t or can’t mount • Get urine often • Poorer quality ejac. • Used in rams and beef bulls

  24. Massage Method • Stimulate by rectal massage • Seminal vesicles • Vas deferens

  25. Semen Evaluation • Appearance (color) • No debris • No puss • No urine • Volume • Motility • Concentration • Hemocytometer, spectrophotometer • Morphology

  26. Preservation of Semen • Extenders (7 components) • Nutrients • Glucose, fructose • Cold shock prevention • Milk, skim-milk, egg yolk • Buffer • Citrate, Tris • Osmotic pressure • The buffer component

  27. Preservation of Semen (cont.) • Inhibit bacterial growth • antibiotics • Increase volume • Cryoprotectant • glycerol

  28. Preservation of Semen (cont.) • Liquid Semen • Collect semen • Semen quality exam • Extend 1:3 (semen:extender) • Minimal extension rate • Cool to 5°C over 2 hours • OK for bull, stallion, ram • Boar - cool to 15°C

  29. Preservation of Semen (cont.) • Once cooled, extend semen to final amount • Bovine (inseminate 0.5 ml) • 2 to 5 million sperm/ml • Equine (inseminate 1 billion sperm) • 25 to 50 million sperm/ml • If don’t cool then inseminate 500 million motile sperm • Swine (inseminate 1.5 to 6 billion sperm in 50 ml) • 30 to 120 million sperm/ml

  30. Preservation of Semen (cont.) • Frozen semen • Follow instruction for collecting and cooling semen • After cooling to 5°C, extend to 2X the final concentration desired • If want final concentration to be 40 million/ml then dilute to 80 million to ml at this time • Hold semen for 4 to 6 hours at 5°C • Equilibrates semen to the cold

  31. Preservation of Semen (cont.) • Add the cryoprotectant • Mix extender with 2X final cryoprotectant amount, 1:1 with extended semen • Do this in small portions to minimize cryoprotectant toxicity • Package semen • 0.5 ml French straws • Ampules • Freeze semen • Liquid nitrogen vapor • Static • Mechanically controlled • Dry ice depressions for pellet freezing

  32. Preservation of Semen (cont.) • Storage • In a liquid nitrogen tank

  33. Liquid Nitrogen Tank

  34. Temperature in Neck of Storage Tank Liquid Nitrogen -196°C

  35. Temperature of Straws If Low LN

  36. Thawing • Use the procedures recommended by the semen supplier!!! • 35°C water for 30 - 60 seconds • Ice water for 3 minutes • Pocket thaw

  37. Insemination of the Female • Detection of estrus • No need to review this material • Time of insemination • Cattle (2X daily heat detection) • 12 hours after observed in standing heat (AM - PM rule) • Inseminate on the day of estrus • Swine (2X daily heat detection) • Sow - 24 and 36 hours after first seen in estrus • Gilt - 12 and 24 hours after first seen in estrus

  38. Insemination of the Female • Sheep • 12 to 18 hours after first seen in estrus • Horses • Every second day beginning on day 3 of heat • Breed when reach 40 - 45 mm follicle • Breed 24 hours after HCG injection • HCG given when a >35 mm follicle is present • Ovulation is 36 to 40 hours after HCG • Insemination protocol • Rectal-vaginal • Vaginal

  39. Insemination of the Cow Rectal-Vaginal Approach

  40. Artificial Insemination in the Mare Vaginal

  41. Artificial Insemination in the Sow

  42. Factors Effecting Conception Rate • How is conception rate measured? • Non-return rate • Rectal palpation • Ultrasound • Time of insemination • If after ovulation then get aging of oocytes • # of sperm inseminated • Fertility of males • Skill of inseminator

  43. Use and Success of AI Semen Species Liquid Frozen Preg. Rate Major Problems Dairy Cattle OK OK 60-70 OK, need good heat detection Beef Cattle OK OK 55-65 Range area large: poor heat detection Sheep OK Fair 50-65 Large range; low value of ewe Swine OK Fair 40-75 Estrus detection Horses OK Fair 30-60 Timing insemination, breed restrictions Turkey OK Poor 90 None Humans OK Fair 5-30 Donors; infertility; time

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