Instructions for thawing and Worksheet

Cooling/Freezing Worksheet

Collection Date 8/8 2006

Antibiotic: Ticarcillin

Tank Owner: Allendale Farms LLC

EVA  Test: Negative

Sperm per ml: 295

Total Raw ml: 53

Devided by Std Dose: 500

% Motility(Est) 75.00%

% Motility after Freeze (Est): 50.00%

Processing Loss 3%: 15.17

Total Frozen Doses: 15.17

Breeding Instructions For Frozen Semen Processed By Allendale Farms LLC

Step 1 - The Mare

The Mare must be monitored throughout her heat cycle for follicle growth.  When ovulation is immanent (38-44 mm), mare should be checked every 6-12 hours to detect ovulation.  When ovulation has been confirmed, the semen straws should be thawed, and inseminated.
DO NOT breed prior to ovulation.

Step 2 - Thawing the Semen Straws

Confirm the cane ID and the straw color on your Frozen Semen Work Sheet.  Also, check the number of straws per dose.  Prior to retrieving the straws from the liquid nitrogen tank, you will need to prepare two (2) water baths.  The 1st bath should be 45 Degrees C and the 2nd bath should be 37 Degrees C.  Thaw one (1) straw at a time.  The straw should be removed from the liquid nitrogen tank and immediately put into the 1st water bath for 10 seconds.  Hold the straw under the water (straw will try to float).  Immediately place the straw in the 2nd water bath for 30 seconds minimum.  NEVER let any straws be exposed to outside air temperatures (raised above the frost line in the liquid nitrogen tank) for over 4 seconds unless it is being thawed.  Four (4) seconds should give you ample time to retrieve the straw to be thawed.  After thawing, shake the air bubble to the sealed end.  DO NOT cut the wick end of the straw. 

Step 3 Artificial Insemination of the Mare 

IMMEDIATLY after thawing the straws, artificially inseminate (AI) the mare.  Procedure #2 may be more practical.

Procedure 1:

Deep uterine insemination: Use 1/2 ml straw AI pipette.  (Flexible models are available through Mintube of America 1-800-646-4882, style # 17209/1075, pipette # 17209/1275.  It is important the tip of the pipette is in the uterine horn toward the ruptured follicle. 

Procedure 2:

Use 8-10 ml Next Generation Basic Formula Extender, or equivalent (no antibiotics).  Empty all thawed straws into the extender.  Mix by inversion.  DO NOT SHAKE.  Use a standard equine AI pipette and a ALL PLASTIC syringe.  Flush the AI tube with 4 ml of plain extender to wash all semen through the pipette.
IMPORTANT: EXTENDER MUST BE THE EXACT TEMPERTURE AS THE STAWS WHEN MIXING.

Suggestions

Do not use abnormal mares.

AI all semen into the uterine horn toward the ruptured follicle.  (flexible pipette).

It may be difficult to check motility using AI procedure.  In the unlikely event your tank loses all of it's liquid nitrogen, discard every straw in the tank.  Keep your tank in plain view.  Ice or frost should not form on outside of tank.  If you suspect a tank failure, move semen to another tank immediately.  A thawed straw cannot be refrozen.

Keep your liquid nitrogen tank, work sheets and these instructions in a safe place.

Do not allow liquid nitrogen tanks to come in direct contact with concrete, asphalt or another tank.

These instructions are from information purposes only, and should not be used in some states.  Tennessee code annotated 63-12-103/9. a. (1)  Kentucky revised statues 321.181.5 .a.

If you have any questions, contact Allendale Farms LLC at 706-476-5415