1. 1. Kristin Block, MANGA Licensed InseminatorMandy Lockard, RVT
2. 2. Introduction Kristin Block Owner of Block Sporthounds, Inc. Breeding and raising greyhounds for over 15 years NC State Licensed Laboratory NGA Stud Master NGA Licensed Inseminator Now Freezing AKC Semen
3. 3. Training and Experience Trained under Sharon Conole of Camelot Farms inCollege Station, TX 80 hours of intensive training in one week coveringcollection, assessing motility and morphology and theart of freezing Has been freezing semen for nearly 15 years
4. 4. NGA Requirements ForFreezing Semen Must complete three test freezes that are inspectedand approved by NGA A third party has to use your semen to impregnate abitch with a resulting litter
5. 5. NGA Requirements forLicensed Inseminator Inseminators must use semen from another freezingfacility that results in a litter of pups All breeding data is reported and peer reviewed onlineat ngagreyhounds.com
6. 6. Terminology Artificial Insemination or AI anything other than livecoverage; most commonly refers to semen beingdeposited in the vaginal canal Trans Cervical Insemination or TCI A non-surgicalendoscopic procedure where a camera visualizes thecervix, a small catheter is inserted through the cervixand semen is deposited into the uterus Surgical Insemination A surgical procedure wherethe abdomen is opened, uterus located and semendeposited into the uterus
7. 7. Terminology Fresh or Fresh Chilled Semen Fresh out of theshoot or placed on extender, chilled and shipped;semen that has not been frozen Extended Semen refers to preparing the semen witha special fluid, or media, that is added to the spermcells to help keep them healthy. Extender is used when shipping the semen, and oftenwhen preforming an AI or TCI with fresh semen A different type of extender is used during the freezingprocess
8. 8. Artificial Insemination orNatural Breeding? The single biggest advantage of artificial breeding isyou know what is going in the bitch Less change of the bitch or dog getting hurt Reduces chances of spreading STDs to the stud dog It is recommended that the bitch owner requires abrucellosis test from the stud dog
9. 9. Preparing for the CollectionHomework for the Human Health Testing Not necessary, but recommended Paperwork DNA Only needed when freezing Most registries are requiring DNA for freezing (AKC, UKC) Registration Papers Not necessary if you are just wanting to see if your boyhas swimmers Testicle Check
10. 10. Preparing for the CollectionThe Stud Dog Food Intake No large meals before collecting Walk him and let him urinate after he arrives at the collection center
11. 11. The Collection Should take place in a stress free location withminimal distractions Clear disposable cones should always be used Teaser bitch will give you a better collection Only Sperm Rich Fraction (SRF) is needed The SRF is about 2-3 mls and cloudy in color Prostatic fluids, which contain no sperm cells, can reachvolumes of 20+ mls. Prostatic fluids are normally clear, although blood iscommonly seen
12. 12. Semen Evaluation Semen should be immediately evaluated for motility on warm slides Cold slides will cool the cells and decrease the motilityresulting in improper assessment Even in a side-by-side AI, semen should be evaluated as it is possible for a collection to look as though there is semen, when in fact, there is not!
13. 13. Semen Example
14. 14. BSI Equipment BSI uses a Phase Contrast Microscope, which allows for 3-D imaging of the cells Left to right: Phasecontrast microscope,slide warmer, variablespeed centrifuge,water bath
15. 15. Semen Evaluation Sperm cells need to be evaluated for abnormalities Abnormal cells cannot impregnate an egg Only normal cells are counted when determining a breeding unit, and thus why the evaluation is vital If you have 1 billion abnormal cells, you have NO breeding units!! Proper equipment is needed to appropriately evaluate the sperm cells
16. 16. Normal Sperm Cell
17. 17. Examples of Abnormal Cells
18. 18. What to do with semen Up to this point, the semen can be used for: Side-by-side AI Chilled, extended and shipped Frozen We will be discussing frozen semen Feel free to ask questions about fresh though!
19. 19. Frozen Semen Advantages You immortalize your dog No relying on the dog to perform, whether it benatural breeding, collecting for a side-by-side orshipping Multiple bitches can be bred at the same time inmultiple locations Dogs from all over the world can be used Your dog can be sent anywhere in the world
20. 20. Selecting a Freeze Center Franchises are popping up everywhere Just like most franchises, each is individually owned and operated Quality may vary from place to place Check out the equipment. Remember: Bright field microscopes are most common, but are not vey good for properly assessing morphology. Phase contrast is preferred. Semen should be evaluated when warm. Cold slides, pipettes and covers slips will shock the cells.
21. 21. Selecting a Freeze Center Respect anyone who tells you the semen isnt good andnot freezable Never Just go ahead and freeze if the semen is poor Think of the bitch owner that purchases the semen,spends valuable time and money to breed their bitch tofind out the semen isnt good enough to get herpregnant Same goes for you if you want to use it later down theline!
22. 22. After the Freeze You have decided to freeze You choose your freezing center You now have frozen breeding units What does that mean???
23. 23. Breeding Units A breeding units contain 100 million live, normal, motile cells post thaw This is the minimum accepted number of cells it take toget a bitch pregnant Breeding units are most commonly frozen in vials orstraws The total number of sperm cells does NOT indicatehow many breeding units you will get Motility and abnormal cells have to be compensated for
24. 24. Straws Post that recovery is about 50% Breeding units generally consistof 2-6 straws Detailed records must be kept toknow how many straws are abreeding unit from each freeze One straw must be thawed tocheck post thaw motility That could be a breeding unit!
25. 25. Pellets Post thaw recovery is about 70% Pellets can be different colorsand sizes They are packaged in a vial One vial equals a breeding unit The number of pellets in abreeding unit can vary Only one pellet is needed forpost thaw motility check
26. 26. Storing Your Semen Semen is stored in liquidnitrogen tanks at -330 F Electricity is not needed tooperate the tank, but tanks needto be filled on a regular basis Tanks can fail you should carryinsurance on your semen if it isextremely valuable to you
27. 27. Moving Your Semen Semen cannot thaw duringshipment, so vapor shipper isneeded Correctly prepared dry/vaporshippers do not contain free liquidnitrogen. Liquid nitrogen is a regulatedUSDOT Hazardous Material A properly prepared dry/vaporshipper is not. This allows formore flexibility and less cost whenshipping non-hazardous frozenmaterials.
28. 28. Moving Your Semen Most vapor shippers are sent overnight or 2-day FedEx Vapor Shippers are expensive and must be returned to the owner, so when shipping, there is also the cost to return the shipper to the owner Return is less expensive as it can be sent ground ratherthan express
29. 29. Using Frozen Semen Frozen semen must be surgically implanted or transcervically inseminated Both require progesterone testing, however TCI ismuch safer in the hands of a skilled inseminator Vaginal AI with frozen semen is discouraged as theconception rate drops to less than 30%
30. 30. Reproductive Anatomy OvaryUterine HornsCervixVaginal Vault
31. 31. TCI TCI is performed while your bitch is awake andstanding with you by her side if you want to watch atBlock Sporthounds, Inc. Using a camera, the cervix is visualized and a smallcatheter is inserted through the cervix into the uterus You can actually watch the semen being deposited intothe uterus on the TV monitor Most bitches take only a few minutes to inseminate Very few bitches cannot be catheterized
32. 32. Myths and Facts Myth: Frozen Semen must be surgically implanted Fact: TCI is as effective as surgical when properly preformed Myth: Frozen semen shouldnt be used on a maiden bitch Fact: There is no reason why frozen semen cant be used on a maiden Consideration should be taken if the semen is extremely valuable, as your maiden may have unknown reproduction issues
33. 33. Legal EducationNorth Carolina Veterinary Practice Act Article 11, Chapter 90 90-181. Definitions (6) "Practice of veterinary medicine" means: a. To diagnose, treat, correct, change, relieve, or prevent animal disease, deformity, defect, injury, or other physical or mental conditions; including the prescription or administration of any drug, medicine, biologic, apparatus, application, anesthetic, or other therapeutic or diagnostic substance or technique on any animal. Our Lay Interpretation if you do anything - such as a vaccine, pull blood or microchip to an animal that you dont own you are practicing veterinary medicine without a license. However: 90-187.10. Necessity for license; certain practices exempted. Nothing in this Article shall be construed to prohibit: (6) Any person from providing for or assisting in the practice of artificial insemination
34. 34. AKC Education Currently AKC only allows Veterinarians to dobreedings with frozen semen, although they will allowanyone to do a breeding with fresh semen. UKC does not regulate who works with frozen semen