Lecture 14: Sexual Behavior (cont.) B. Applied Reproductive Behavior of the Male: Semen Collection and Processing Text: Ch. 10 and 11Age When Semen Can Be CollectedEffect of Age on Sperm OutputSemen CollectionCollection on a Live MountSlide 6PowerPoint PresentationSlide 8Collection of a Bull on a DummyStallion Collection on a PhantomBoar Collection on a DummyRam Collection on a DummySemen Collection (cont.)Slide 14Effect of Collection Frequency on Sperm OutputArtificial VaginasCharacteristics of AVElectroejaculationMassage MethodSemen EvaluationPreservation of SemenPreservation of Semen (cont.)Slide 23Slide 24Slide 25Slide 26Slide 27Slide 28Liquid Nitrogen TankTemperature in Neck of Storage TankTemperature of Straws If Low LNConclusions on LN TankThawingLecture 14: Sexual Behavior (cont.)B. Applied Reproductive Behavior of the Male: Semen Collection and ProcessingText: Ch. 10 and 11Lecture 14: Sexual Behavior (cont.)B. Applied Reproductive Behavior of the Male: Semen Collection and ProcessingText: Ch. 10 and 11Animal Science 434Animal Science 434Age When Semen Can Be CollectedBull 12 monthsBoar 6 - 8 monthsRam 6 - 9 monthsStallion 20 - 24 monthsDog 8 - 12 monthsEffect of Age on Sperm OutputSperm OutputAge in Months10 20 30 40 50 600BullBullSemen Collection•Sexual arousal»Sight, sound, smell, touch»Best mount - LiveCollection on a Live MountCollection on a Live MountCollection on a Live MountCollection on a Live MountSemen Collection•Sexual arousal»Sight, sound, smell, touch»Best mount - Live»Alternative mount - dummies•Bull•Stallion •Boar•RamCollection of a Bull on a DummyStallion Collection on a PhantomStallion Collection on a PhantomBoar Collection on a DummyRam Collection on a DummySemen 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 goatsSemen Collection (cont.)•Frequency of collection»As increase frequency/wk •decrease sperm/ejaculate•Increase sperm recovered/wkEffect of Collection Frequency on Sperm OutputItem Dairy Beef Sheep Swine Horses# of collections 1-6 1-6 7-25 2-5 2-6Volume (ml) 5-8 3-6 0.8-1.2 150-300 30-100Concentration(million/ml) 1000-2000 800-1500 2000-3000 200-300 200-400Total sperm/ejac.(billion) 7-15 5-10 1.6-3.6 30-60 5-10Total sperm/wk(billion) 15-40 10-30 25-40 100-150 15-30Motile sperm(%) 50-75 40-75 60-80 50-80 40-75Normal sperm(%) 70-95 65-90 80-95 70-90 70-90Artificial VaginasWaterWaterCollection coneCollection vialInner LinerCharacteristics of AV•Temperature»Bull, stallion, ram - 45°C•Pressure•Friction»Lubrication essentialElectroejaculation•Useful if male won’t or can’t mount»Get urine often»Poorer quality ejac.•Used in rams and beef bullsMassage Method•Stimulate by rectal massage»Seminal vesicles»Vas deferensSemen Evaluation•Appearance (color)»No debris»No puss»No urine•Volume•Motility•Concentration»Hemocytometer, spectrophotometer•MorphologyPreservation of Semen•Extenders (7 components)»Nutrients•Glucose, fructose»Cold shock prevention•Milk, skim-milk, egg yolk»Buffer•Citrate, Tris»Osmotic pressure•The buffer componentPreservation of Semen (cont.)»Inhibit bacterial growth•antibiotics»Increase volume»Cryoprotectant•glycerolPreservation 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°CPreservation 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/mlPreservation 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 coldPreservation 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 freezingPreservation of Semen (cont.)•Storage»In a liquid nitrogen tankLiquid Nitrogen TankTemperature in Neck of Storage TankLiquid Nitrogen -196°CTemperature of Straws If Low LNConclusions on LN Tank•Maintain LN level as high as possible. •Keep extra tank (narrow neck) with only nitrogen so semen storage tank can be refilled when opened. •Enter semen storage tank only when needed •Do not hold canes above neck of LN tankThawing•Use the procedures recommended by the semen supplier!!!»35°C water for 30 - 60 seconds»Ice water for 3 minutes»Pocket thaw•Have thawing container near LN tank when removing
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