Viability of canine spermatozoa is decreased after freezing and thawing. This is due to physical destruction of cells as they undergo osmotic change, dehydration, ice crystal formation, and induction of chemical reactions during the freeze– thaw process that mimics capacitation. Insemination directly into the uterus is strongly recommended. Reported conception rates with frozen – thawed semen for vaginal insemination average 45%, for transcervical insemination 70%, and for surgical intrauterine insemination 95%
Frozen semen is made up of ejaculated spermatozoa suspended in an extender. Extender is a fluid medium containing nutrients, buffers, and anti - microbials and, for frozen semen, a cryoprotectant to minimize disruption of the spermatozoa cells as they freeze and thaw. Cryoprotectants most commonly used are egg yolk and glycerol. Egg yolk may also prevent premature capacitation of spermatozoa.
Collect semen in a routine manner and perform a complete semen evaluation. Only very good - to excellent - quality semen warrants freezing. Samples with high percentages of proximal cytoplasmic droplets may not be suitable for freezing; presence of proximal cytoplasmic droplets has been associated with decreased fertilizing capability and poor survival after freezing and thawing. Semen samples containing 40% or greater blood and blood products are not suitable for cryopreservation; haemoglobin released by haemolysis decreases post - thaw viability.
The semen then goes through a process of extenders and slow cooling before finally being ready to transfer into straws in a permanent tank containing liquid nitrogen. All samples must be maintained in fluid liquid nitrogen or liquid nitrogen vapor for storage or shipment until they are to be thawed for insemination.
The semen of some male dogs does not tolerate freezing, presumably because of a subclinical disease or because of the semen’ s chemical incompatibility with the extender. It is strongly recommended that dogs intended for use as a stud with frozen semen have semen collected and frozen, and one straw or pellet immediately thawed to assess post - thaw quality. This should be done well before any bitches of interest are in heat. Because some males can be identified as having semen that is difficult to freeze, breeders should be reminded that their unique problem may have a genetic basis.
Semen must be maintained in fluid liquid nitrogen or liquid nitrogen vapor at all times. Some shippers refuse to carry canisters containing fluid liquid nitrogen.
“Dry shippers” contain liquid nitrogen vapor and will remain charged for 1 to 3 weeks, permitting shipment; either the dry shipper should be replenished with fluid liquid nitrogen on arrival or the straws transferred to a more secure liquid nitrogen container upon arrival.