The invention relates to a 
livestock semen freezing 
diluent as well as a preparation method and application of the 
livestock semen freezing 
diluent. The preparation method of the 
livestock semen freezing 
diluent comprises the following steps of: adding 2.71 grams of trihydroxymethyl aminomethane, 1,4 grams of citric acids, 1.0 
gram of monosaccharides, 10 myriad IU (
International Unit) of 
penicillium, 10 myriad IU of 
streptomycin and 0.29-6.97 grams of 
inositol compounds to a right amount of 
ultrapure water; optionally adding 5-20 milliliters of penetrability protectants; uniformly stirring; regulating a pH value to 6.8-7.2; adding 5-20 milliliters of fresh 
yolk subjected to inactivation treatment; uniformly mixing, and then fixing the volume to 100 milliliters; centrifugalizing at low temperature for 1 hour; and filtering supernate to prepare the livestock semen freezing diluent. Because the 
inositol compounds can effectively inhibit the generation of 
ice crystals, the livestock semen freezing diluent disclosed by the invention reduces the mechanical injury caused by the 
ice crystals on semens, achieves the 
survival rate of the defrozen semens at about 75%, the activity more than 50%, the 
acrosome integrality at about 60% and the plasmalemma integrality at about 50% and achieves the non-return rate more than 70% after 
artificial insemination; and the livestock semen freezing diluent has the advantages of no immune 
response generation, purity guarantee, low price, and the like by using the 
inositol compounds as the ice 
crystal inhibitors of the semen freezing diluent.