Artificial Insemination AI What Is AI General definitionthe

Artificial Insemination AI

What Is AI? • General definition-the introduction of semen into the oviduct or uterus by some means other than sexual intercourse. • Ag definition- the use of semen from a genetically superior male to inseminate a female resulting in a genetically superior offspring.

Where Is AI Used? • • Cattle Horses Swine Sheep Goats Dogs Humans • • Turkey Chickens Rabbits Fish Fox Mink Bees

Why Use AI? • AI provides the producer an opportunity to use sires possessing superior genetics. Depending upon the needs and goals of an individual's breeding program, AI offers an economically feasible means of increasing productivity over a wide range of traits. • In spite of this enormous potential to improve production levels, many producers have not put themselves in a position to take advantage of the benefits offered by AI.

Advantages • • • Best possible sires of proven quality Disease control Cost effectiveness Flexibility Safety No bull, stallion, ram, boar, etc.

Overview of Artificial Insemination (What Has To Be Considered First)

Fertility Testing • Important step in evaluating males to be kept for breeding. • Indispensable in the area of artificial insemination since a bull may be worth thousands of dollars & still produce progeny long after his death (because the semen is stored). • The process of evaluating the bull's ability to reproduce.

Factors to consider in Testing • Libido (sex drive) - This is especially important in bulls kept for natural breeding since some bulls are more active breeders than others. • Scrotal circumference - This is an observable, measurable characteristic that indicates fertility. • In general, the greater the scrotal circumference, the greater the sperm production, hence increased fertility. • Semen evaluation - This is the most important factor in fertility testing. Semen is evaluated for:

Semen Evaluated For Appearance • A uniform solid, dense appearance indicates a high sperm concentration. • A sample which appears to be translucent contains fewer sperm cells. • Semen with a curd appearance should not be used as this indicates inflammation.

Semen Evaluated For Volume • May vary depending on the age of the bull or sire. • Younger sires produce a smaller volume of semen which is not a negative factor as long as the concentration of semen is good.

Motility • It must be evaluated under a microscope at 400 X. • Most semen should contain 70 percent or more motile cells

Concentration • Determined by the number of sperm per milliliter of semen. • This is probably the most important indicator of fertility, except with sires, in which scrotal circumference is the determinant.

Morphology • The proportion of abnormal sperm should not exceed 20 percent. • Most males do, however, produce some abnormal sperm.

Semen Collection Methods: Artificial Vagina in goats and cattles • Consists of an outer tube or casing which is constructed of heavy rubber or plastic, and an inner tube or lining of thin rubber. • The space between the two tubes is filled with warm water, which maintains the collected sperm at a constant temperature, neither too hot or too cold.

Semen Collection Methods: Artificial Vagina • One end of the apparatus is open to allow the entrance of the penis and the other end is attached to a glass tube or beaker to receive the ejaculated semen. • This is the most commonly used device for collecting semen.

Semen Collection Methods: Electric Stimulation • An electro-ejaculator apparatuses introduces a weak alternating current to the sacral and pelvic nerves via electrodes placed in the rectum until erection and ejaculation occur.

Semen Collection Methods: Electric Stimulation • It is used on all farm animals except boars and horses. • It is used effectively on bulls that cannot mount due to leg injuries.

Semen Preparation: Storage & Shipment • It may be kept at room temperature if it is to be used within two hours of collection. • If the semen will be put into long-term storage, semen needs to be gradually cooled and frozen a temperature below zero.

Semen Preparation: Semen Extenders • Why Added? • Add extra needed Volume • Exert beneficial effect on the sperm

Semen Preparation: Semen Extenders • 1. Egg yolk – phosphate: used in bull, ram, and stallion semen • 2. Egg yolk – citrate: used in bull and ram semen • 3. Homogenized whole milk: used in bull, ram, and boar semen

Semen Preparation: Semen Extenders cont. • 4. Glycine-containing diluents: used mostly in boar semen. • 5. Some extenders contain antibiotics to control bacteria and contamination.

History • It may have been used as far back as 1322, at which time an Arab chieftain used artificial methods to impregnate a prized mare. • The first scientific research relative to A. I. in domestic animals was conducted with dogs by the Italian physiologist Lazarro Spallanzani.

Equipment Needed For A. I. CATTLE GOATS AND HORSES • 1. Liquid nitrogen tank – Used to store semen straws at -320 degrees Fahrenheit – Semen stored in tank, may last indefinitely once froze properly & temperature is maintained

Equipment Needed For A. I. cont. • 2. A semen straw – containing a single dose of semen • 3. Straw tweezers – Used to pick up the semen when in the tank & thawing unit

Equipment Needed cont. • Semen Thawing Unit – Used for thawing the frozen semen before insemination – Contains water • Thermometer – To measure the temperature that is in the thawing unit – Temperature should be between 95 – 98 degrees Fahrenheit

Equipment Needed cont. • Inseminating syringe or gun – Made from stainless steel – Used to place semen in the reproductive tract of the cow or heifer and to maneuver it through the cervix • Sterile Lubricant (Lube) – Lubrication put on plastic sleeve in order to first enter the vulva

Equipment Needed cont. • Paper Towels – To dry off the semen straw – To wipe animal clean, especially the vulva – To put on top of the vulva to keep clean • Scissors – To cut the end of the straw just prior to insemination

Equipment Needed cont. • Plastic Sleeve – To put on inseminator’s arm to keep clean • Cover Sheath – Inseminator’s gun is put inside of the sheath so that all is kept clean

Equipment Needed cont. – Ka. Mar heat strips: heating aid with red dye that expels when a cow or heifer is mounted – Tail Chalk: chalk paint is smeared when cow or heifer is mounted

Disadvantages • Equipment- availability • Time – Careful herd observation required • 3 -4 observations per day – 15 -20 minutes per observation

Steps to AI (Semen Storage) • Collected, evaluated, cooled slowly, & frozen – -320°F • Lasts several months • 30 years • 40 years

Steps to AI (Heat Detection) • • • Increased activity Mounting Swelling and redness of vulva Discharge Winking – Mares • Standing heat – Best indicator of estrus

Steps to AI (Insemination) • Cattle – 12 hours after heat detection • Horses – 3 rd, 5 th, & 7 th day of estrus Horses • Swine – 24 and 12 hours after onset of estrus in sows – 12 and 24 hours after onset of estrus in gilts • Sheep – Two inseminations increase conception and multiple lambs

Recto-vaginal Method (Cattle) • Remove all feces from the rectum • Grasp the cervix through the wall of the rectum – Not done with horses or swine • Insert inseminating tube through vagina and into the cervix • Guide tube through cervix • Deposit semen from middle of cervix to body of uterus

Recto-vaginal Method

Lapriscopic Artificial Insemination (Sheep) • Semen injected from a syringe through a pipette into upper one-third of uterine horn through abdominal wall

Lapriscopic Artificial Insemination

Artificial Insemination procedures • Step #1: Restrain the animal to be inseminated. Step #2: • Raise the tail with the right hand gently massage the rectum with the lubricated glove on the left hand. • Step #3: Gently wipe the vulva with a paper towel to remove excess manure and debris.

Procedure for artificial insemination Step #4: Insert the gun at a 30° upward angle to avoid entering the urethral opening and bladder located on the floor of the vagina.

Procedure for artificial insemination Figure #1: Keeping the gloved hand even with the tip of the inseminator gun. Figure #2: Allowing manure to pass over the top of the hand arm.

Procedure for artificial insemination Figure #3: Dealing with colon constrictions. Figure #4: Grasping the cervix and gently moving it forward

Procedure for artificial insemination Figure #5: Close-up of the cervix. Figure #6: Finding the opening of the cervix.

Procedure for artificial insemination Figure #7: Moving the cervix over the tip of the insemination gun. Figure #8: Locating the end of the insemination gun.

Procedure for artificial insemination Figure #9: Depositing the semen in the body of the uterus Figure #10: Good distribution of the semen to both uterine horns.

Procedure for artificial insemination Figure #11: Improper distribution of the semen into one horn because the insemination gun is pushed too far forward.

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 Updated: 11/22/2020

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: 11/22/2020

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: 11/22/2020

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: 11/22/2020

Age When Semen Can Be Collected Bull Boar Ram Stallion Dog Updated: 11/22/2020 12 months 6 - 8 months 6 - 9 months 20 - 24 months 8 - 12 months

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: 11/22/2020

Preservation of Semen (cont. ) – Inhibit bacterial growth • antibiotics – Increase volume – Cryoprotectant • glycerol Updated: 11/22/2020

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: 11/22/2020

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: 11/22/2020

Preservation of Semen (cont. ) • Frozen semen – Follow instruction for collecting and cooling semen – After cooling to 5°C, extend to 2 X 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: 11/22/2020

Preservation of Semen (cont. ) – Add the cryoprotectant • Mix extender with 2 X 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 Updated: 11/22/2020 – Mechanically controlled

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 Humans Updated: 11/22/2020 OK Fair 5 -30 None Donors; infertility; time

• • References: 1. Dev. Biol- Verma and Agarwal 2. Dev. Biol- Wolpert 3. Dev. Biol. - Gilbert 4. Embryology- Ballinsky 5. Wikipedia 6. Freely available resources on internet