Clone AI Kits

GUIDELINES FOR USING FRESH CHILLED EXTENDED SEMEN KITS

• Start preparation well in advance, if you think there is a possibility of shipping semen in the future, order a kit and keep it in the freezer.
• Bitch owner needs to find out the Ministry of Agriculture and KC requirements regarding import of chilled semen into their country
• Stud dog owners needs to find out any DEFRA requirements regarding export of chilled semen from the UK, contact local DEFRA office.
• Bitch owner needs to use a vet who has suitable ovulation test and is willing to Inseminate at any time. (Chilled semen needs to be inseminates when a bitch is in oestrus and this needs to be established using an ovulation test)
• Stud dog could be collected by owner or vet, some KCs will ask for a veterinary note confirming collection from a specific stud dog (microchip or DNA profile) so your vet needs to co-operate.
• You may wish to practise collecting semen the stud dog in advance if you are not familiar with this.
• Most dogs require an in season bitch as a tease in order to make a satisfactory collection, especially if they are not used to the procedure.
• A kit can be sent to you Using Royal Mail Special Delivery, which Guarantees delivery by 1pm the next day, be sure that someone can receive and sign for this.
• Put a kit (or the two phials & ice pack) into your freezer until you need to use them. * Cost of kit is £55 plus P&P (currently £7.75) paid in advance, total - £62.50
• Kits containing chilled semen can be exported to most major destinations Provided that time from collection to insemination is less than 36 to 48 hours and temperatures remain between 5-15 degrees Celsius

SHIPPING OF FROZEN SEMEN
OR
IMPORTATION AND INSEMINATION OF FROZEN SEMEN

Please contact the UK Veterinary Representative for Clone

CLONE - CHILLED SEMEN PREPARATION

KEEP VIALS OF SOLUTION & ICE PACKS FROZEN UNTIL READY TO USE

(Vials may be thawed but cold when received, just refreeze immediately)

1. Thaw the FROZEN EXTENDER VIAL (orange or salmon colour vial) It must be at room temperature when it is added to the semen after you have collected it.
2. Assemble the lastic collection bag onto the centrifuge tube.
3. Tease the stud with an estrus bitch if possible. When the stud has full erection, let let two squirts of first fraction be expelled to wash out the urethra before collecting the sperm portion.
4. Collect the sperm rich fraction only, stopping when you notice any clear fluid being ejaculated.
5. If you collect in the plastic bag, let the semen run into the corner of the bag, then cut of the other bottom corner of the bag and gently pour into the centrifuge tube.
6. Ask the bitch owner how the insemination is going to be performed?  You must know how the insemination is to be done to prepare the semen for shipment. Check the Extender Addition Volumes on the form. If a Trans-Cervical or surgical insemination is to be done, you must have a condensed pellet of semen so you have a final insemination volume as on the table enclosed.
7. If Step #4 above can't be accomplished without collecting a large amount of prostate fluid, you must then centrifuge the semen at urine speed (700 g; very slow) for 3-4 min. Draw off the prostate fluid with a syringe very slowly, leaving about a quarter to half a ml of prostate behind. There should be no more fluid than .50 ml for small dogs and 6-7 mls for large dogs to add to the extender for a regular insemination and no more than .25 and 1ml to be add to the extender for a Trans-cervical or surgical insemination.
8. Re-suspend the sperm cells into the remaining prostate fluid by gently rocking the centrifuge tube until the sperm cells are totally back into the solution.
9. Add the right amount of orange or salmon coloured extender to semen, slowly.
10. Attach the centrifuge tube to its Styrofoam holder with a rubber band. (You could use 4 wraps of paper towels in lieu of the styrofoam holder)
11. Set the frozen ice pack next to the styrofoam semen holder, keeping the tube of semen from touching the ice pack by facing it away from the ice pack.
12. Make sure the Semen Activator vial (clear solution) Insemination instructions, syringe & pipet are included in the box for the inseminator of the bitch. Tape the box closed and ship by way of Next Day or Faster. Do not use regular Postal Service. The semen will cool on it's way and be ready to reactivate when it arrives at its destination.

Collection Directions

There is now a Plastic Zip Bag for the semen collection that replaces the old plastic cone or funnel.

1. Have a sharp scissors on hand.
2. At the top of the zip bag or pull off the sealed closure at the perforation just above the yellow wire band, (if you have one.)
3. Open the bag and fold back the yellow tabs that stick out from the bag along side of the bag.
4. Now fold or roll down over the outside of the bag, the yellow tab with the wire stiffener one or two times like you would roll up the cuffs on your shirt at your wrist. This will make a softer entrance into the bag by covering the wire stiffener with more plastic of the bag and will keep the bag open.
5. When collecting the semen, make sure the dogs penis does not go into the bag more than half way as not to contaminate the lower portion of the interior sides of the bag from mucus and bacteria from the penis. this mucus and/or contaminate will stick to the sides of the bag, but will not run down into the semen you collected if you are careful.
6. After collecting the right amount of semen, hold the bag in front of you at eye level and let all the semen run into one of the two corners of the bottom of the bag. Never let the semen above the lowest portion of the bag so it does not pick up any contamination from where the penis was during the collection.
7. With a pair of Scissors cut off on the diagonal the other corner of the bottom of the bag and pore all the semen into the orange covered plastic centrifuge tube.
8. Add the thawed orange or salmon extender to the semen slowly and cap.
9. Attach the centrifuge to to the styrofoam holder with a rubber band
10. Set the frozen Ice pack next to the semen holder. keeping the semen from touching the ice pack by facing it away from the ice pack.
11. Put Semen activator vial (clear solution, Insemination instructions, syringe and pipet in the box and tape the box closed ready for shipping.

INSEMINATION INSTRUCTIONS

CLONE CHILLED SYSTEMS

1. When the semen arrives, keep refrigerated (not it the freezer) until ready for use (at this point the semen is in a almost motionless state)
2. One half to one hour before insemination bring the semen and activator (the clear or blue solution vial) out of the refrigerator and let them both come up to room temperature, and until they are both the same temperature
3. Prepare a water bath of approximately 1 gallon of water, in a container such as Styrofoam box or bucket. Have the water or bath pre-warmed to 85-90 deg F.
4. Add the activator solution slowly to the semen which is in the centrifuge tube at this time (when both are the same temperature)
5. Put the cap back on tightly, then gently rock the semen tube back and forth a few times very slowly until all the solutions are mixed.
6. It's a good idea at this point to lower the semen tube so that the semen portion is submerged in the warm 85-90 degree water bath for 8-10 minutes to activate the semen to almost normal temperature, speed and activity. At this point you may examine the semen to check mobility and forward speed of progression.
7. Assemble the breeding tube or rod to the syringe and draw the semen up the breeding tube into the syringe in one column with no air bubbles.
8. Inseminate the bitch in the elevated position, and keep her rear elevated for 8-10 minutes after the insemination is performed.

NEVER WARM THE SEMEN ABOVE THE TEMPERATURE OF 90 DEGREES FAHRENHEIT. INSEMINATE THE SEMEN AS SOON AS YOU REMOVE IT FROM THE WATER BATH. INSEMINATE AT ROOM TEMPERATURE AND NOT IN ANY COLD AREA AFTER THE SEMEN HAS BEEN PREPARED AND WARMED