Artificial Insemination John Parrish Updated:9/19/2018

Objectives of Artificial Insemination Genetic improvement of livestock Disease control mechanism Possible to increase fertility Decrease breeding expense Updated:9/19/2018

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 Updated:9/19/2018

Advantages of AI Genetic Improvement Rapid proof of sire 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 Updated:9/19/2018

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 Updated:9/19/2018

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 Updated:9/19/2018

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 Updated:9/19/2018

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 Updated:9/19/2018

Effect of Age on Sperm Output Bull Sperm Output 10 20 30 40 50 60 Age in Months Updated:9/19/2018

Semen Collection Sexual arousal Sight, sound, smell, touch Best mount - Live Updated:9/19/2018

Collection on a Live Mount Updated:9/19/2018

Collection on a Live Mount Updated:9/19/2018

Semen Collection Sexual arousal Sight, sound, smell, touch Best mount - Live Alternative mount - dummies Bull Stallion Boar Ram Updated:9/19/2018

Collection of a Bull on a Dummy Updated:9/19/2018

Stallion Collection on a Phantom Updated:9/19/2018

Boar Collection on a Dummy Updated:9/19/2018

Ram Collection on a Dummy Updated:9/19/2018

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 Updated:9/19/2018

Semen Collection (cont.) Frequency of collection As increase frequency/wk decrease sperm/ejaculate Increase sperm recovered/wk Updated:9/19/2018

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 Updated:9/19/2018

Artificial Vaginas Inner Liner Collection cone Collection vial Water Updated:9/19/2018

Characteristics of AV Temperature Pressure Friction Bull, stallion, ram - 45°C Pressure Friction Lubrication essential Updated:9/19/2018

Electroejaculation Useful if male won’t or can’t mount Get urine often Poorer quality ejac. Used in rams and beef bulls Updated:9/19/2018

Massage Method Stimulate by rectal massage Seminal vesicles Vas deferens Updated:9/19/2018

Semen Evaluation Appearance (color) Volume Motility Concentration No debris No puss No urine Volume Motility Concentration Hemocytometer, spectrophotometer Morphology Updated:9/19/2018

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 Updated:9/19/2018

Preservation of Semen (cont.) Inhibit bacterial growth antibiotics Increase volume Cryoprotectant glycerol Updated:9/19/2018

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 Updated:9/19/2018

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 Updated:9/19/2018

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 Updated:9/19/2018

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 Updated:9/19/2018

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Preservation of Semen (cont.) Storage In a liquid nitrogen tank Updated:9/19/2018

Liquid Nitrogen Tank Updated:9/19/2018

Temperature in Neck of Storage Tank Liquid Nitrogen -196°C Updated:9/19/2018

Temperature of Straws If Low LN Updated:9/19/2018

Thawing Use the procedures recommended by the semen supplier!!! 35°C water for 30 - 60 seconds Ice water for 3 minutes Pocket thaw Updated:9/19/2018

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 Updated:9/19/2018

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 Updated:9/19/2018

Insemination of the Cow Rectal-Vaginal Approach Updated:9/19/2018

Artificial Insemination in the Mare Vaginal Updated:9/19/2018

Artificial Insemination in the Sow Updated:9/19/2018

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 Updated:9/19/2018

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 Updated:9/19/2018