Successful ram semen cryopreservation with lyophilized egg yolk-based extender.
The aim of this study was to evaluate the effects of lyophilized egg yolk extender on ram semen cryopreservation. Ejaculates with a thick consistency, rapid wave motion (3-5 on a 0-5 scale) and >75% initial motility were pooled. Sperm were diluted to final concentration of 1/5 (semen/extender) in lyophilized egg yolk or fresh egg yolk extenders using two-step dilution method. The equilibrated semen was frozen in 0.25 mL straws. Semen samples were assessed for sperm motility, plasma membrane functional integrity using hypoosmotic swelling test (HOST), damaged acrosome using FITC-Pisum sativum agglutinin (PSA-FITC) and DNA integrity using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) at three time points: after dilution with extender A, equilibration and post-thaw. The results showed that freezing and thawing procedures (dilution, equilibration and thawing) had negative effects on motility (P<0.001), plasma membrane integrity (P<0.001), acrosome integrity (P<0.001) and DNA integrity (P<0.001). In the study, there were no significant differences between lyophilized and fresh egg yolk extenders when comparing motility, plasma membrane integrity, acrosome integrity and DNA integrity between groups. In conclusion, lyophilized egg yolk extender provided similar cryoprotective effects with fresh egg yolk extender to cryopreserve ram semen.
Department of Reproduction and Artificial Insemination, Uludag University, Veterinary Faculty, 16059 Bursa, Turkey. Electronic address: salcay@uludag.edu.tr.
Selim
S
Alcay
Department of Reproduction and Artificial Insemination, Uludag University, Veterinary Faculty, 16059 Bursa, Turkey.
M
M
Berk Toker
Department of Reproduction and Artificial Insemination, Uludag University, Veterinary Faculty, 16059 Bursa, Turkey.
Elif
E
Gokce
Department of Reproduction and Artificial Insemination, Uludag University, Veterinary Faculty, 16059 Bursa, Turkey.
Burcu
B
Ustuner
Department of Reproduction and Artificial Insemination, Uludag University, Veterinary Faculty, 16059 Bursa, Turkey.
N
N
Tekin Onder
Department of Reproduction and Artificial Insemination, Uludag University, Veterinary Faculty, 16059 Bursa, Turkey.
Hakan
H
Sagirkaya
Department of Reproduction and Artificial Insemination, Uludag University, Veterinary Faculty, 16059 Bursa, Turkey.
Zekariya
Z
Nur
Department of Reproduction and Artificial Insemination, Uludag University, Veterinary Faculty, 16059 Bursa, Turkey.
M
M
Kemal Soylu
D000177:Acrosome / Q000187:drug effects; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004247:DNA / Q000235:genetics; D004530:Egg Yolk / Q000378:metabolism*; D005612:Freeze Drying; D005615:Freezing; D006801:Humans; D020287:In Situ Nick-End Labeling; D008297:Male; D012661:Semen / Q000187:drug effects / Q000502:physiology*; D012662:Semen Preservation / Q000379:methods*; D012756:Sheep / Q000502:physiology*; D013081:Sperm Motility / Q000187:drug effects*; D013094:Spermatozoa / Q000187:drug effects
D003451:Cryoprotective Agents; D004247:DNA
Cryopreservation; Egg yolk; Lyophilization; Ram semen