Establishment Of A Laparoscopic-mediated Artificial Insemination For Chickens

In order to solve the problem of incomplete development of vas deferens in male phenotype reversal hens,we try to establish a chicken laparoscopic artificial insemination method for the collection of sperm.The study shows that:(1)First we selected appropriate anesthesia and the sperm cryopreservation method and compared the efficiency of chicken embryos fertilization and hatching in fresh and frozen insemination to study the feasibility of chicken laparoscopic artificial insemination method.Our result showed that the shearing,grinding and incision of chicken testicular tissue is not important for the volume of semen collection from chicken,and the testicular tissue can directly be ground to obtain more volume of sperm.(2)Secondly,after 6 days of continuous egg production by intramuscular anesthesia and sub-intravenous anesthesia,we found that the difference between the two groups was not significant(P > 0.05),but the number of eggs received on the first day after intravenous injection was higher.(3)In the Third experiment,we compared the semen preservation between TL-HEPES,Green reagent,BPSE and TTE at 37 °C,the results showed that the sperm survival rate in BPSE was significantly lower than that in TL-HEPES and Green reagent(P<0.05),and the sperm survival rate was significantly different from TTE(P<0.01)compared to TL-HEPES,Green reagent and BPSE.After 120 min,the sperm survival rate in BPSE was significantly different compared to TL-HEPES and Green reagent(P<0.01).(4)In the Forth experiment,we used TTE as a diluent to compare the survival rate of chicken sperm with glycerol,DMSO and propylene glycol.The results showed that DMSOwas superior to propylene glycol(P<0.05),while,glycerol was better than propylene glycol(P<0.01).(5)In the Fifth experiment,we applied Glycerol and TTE as cryoprotectants.The results showed that the semen freezing equilibrium time was frozen at 10 cm on the liquid nitrogen surface for 10-30 min.The sperm survival rate after thawing was higher,and the best effect was obtained for 15 min.(6)Finally,we compared the fresh semen,fresh frozen semen and testicular sperm through the cloaca and laparoscopic artificial insemination of chicken embryo fertilization rate,hatching rate and brooding rate.When the amount of semen is about 2.0×107,the fresh essence and the testicles are refined.There was no significant difference in the rate of spermatozoa collection through cloaca and laparoscopic artificial insemination(P>0.05)method.In this study,we found that laparoscopic artificial insemination of frozen sperm and testicular semen were slightly better compared to others.Here for the first time,we successfully established and reported a laparoscopic-mediated artificial insemination method for chickens.