Effects of Semen Extender Containing Equex-STM Paste on Post-thaw Motility and Viability of Canine Sperm

Equex-STM paste 첨가 희석액이 개 정액의 동결.융해 후 정자활력 및 생존율에 미치는 영향

  • Published : 2002.03.01

Abstract

This study was performed to investigate the freezomg condition especially focused on extender composition to achieve good post-thaw viability and motility of canine sperm. Semen were collected from 6 male dogs which had been proved to be fertile in the past and were treated for freezing. Equex-STM paste was contained in both the 1st(3%) and the 2nd(7%) diluent and the 2nd diluent was added to the 1st diluent following glycerol equilibration for an hour and a half. To investigate the effect of Equex-STM paste in the extender on post-thaw canine sperm characteristics, the post-thaw viability, motility, and HOS(Hypoosmotic swelling) values were evaluated according to the different composition of extender with or without Equex-STM paste, thawing conditions, and different thawing media added to thawed semen. 1. Canine sperm removed from seminal plasma and frozen )n Sweden extender containing Equex showed higher post-thaw viability, motility, and HOS values than those frozen in the extender containing Equex-STM paste with seminal plasma and those frozen in the extender without Equex and seminal plasma. 2. Canine sperm frozen in Sweden extender containing Equex-STM paste with 5% glycerol showed higher post-thaw viability, motility, and HOS values than those frozen with 3%, 8% glycerol or 5% DMSO. 3. The canine semen frozen in Sweden extender with 5% glycerol and Equex-STM paste showed higher viability, motility, and HOS values when thawed at $70^{\circ}C$ for 8 seconds than when thawed at $37.5^{\circ}C$ for 1 min and at $18-20^{\circ}C$ for 5 min. 4. TFC (tris -fructose-citrate) and PB S (phosphate buffered saline) medium added immediately to thawed canine semen brought better viability, motility, and HOS values for the sperm than those semen added with TGC(tris-glucose-citrate) and no medium. These results indicated that Equex-STM paste in Sweden extender for freezing the canine sperm which were removed from seminal plasma brought good post-thaw viability and motility of canine sperm. Also of the freezing conditions of canine sperm with the same extender containing Equex, the concentration of 5% glycerol, the thawing condition at $70^{\circ}C$ for 8 sec, and TFC and PBS medium added to the thawed semen brought better post-thaw viability and motility of canine sperm than the other conditions used in this study.

Keywords

References

  1. Zuchthygiene v.10 Insemination with frozen dog semen based on a new insemination technique Andersen K
  2. J Dairy Sci v.70 Glycerolation and thawing effects on bull spermatozoa frozen in detergent-treated egg yolk and whole egg extenders Arriola J;Foote RH https://doi.org/10.3168/jds.S0022-0302(87)80194-7
  3. Cryobiology v.31 Osmotic effects on ram and human sperm membrane in relation to thawing injury Curry MR;Watson PF https://doi.org/10.1006/cryo.1994.1005
  4. J Reprod Fert(suppl) v.47 Effects of four different extenders and three different thawing rates on post-thaw viability of dog semen Dobrinsky I;Lulaic;Barth AD(et al.)
  5. J Reprod Pert Hypo-osmotic swelling of dog spermatozoa England GCW;Plunmer JM
  6. PhD Theisis. University of London The cryopreservation of canine semen England GCW
  7. J Reprod Fert Studies on freezing dog spermatozoa: effect of glycerol on motility after thawing Fontbonne A;Badinand F
  8. Cryobiology v.29 Glycerol permeability of human spermatozoa and its activation energy Gao DY;Mazur P;Kleinhans FW;Watson PF;Noiles EF;Critser JK https://doi.org/10.1016/0011-2240(92)90068-D
  9. Meth Enzymol v.56 Properties of detergents Helenius A;McCaslin DR;Fries E;Tandford C https://doi.org/10.1016/0076-6879(79)56066-2
  10. Biol Reprod v.46 Freeze-induced membrane damage in ram spermatozoa is manifested after thawing: observation with experimental cryomicroscopy Holt WV;Head MF;North RD https://doi.org/10.1095/biolreprod46.6.1086
  11. Biol Reprod v.51 Effects of temperature and restoration of osmotic equilibrium during thawing on the induction of plasma membrane damage in cryopereserved ram spermatozoa Holt WV;North RD https://doi.org/10.1095/biolreprod51.3.414
  12. Teriogenology v.41 Effects of storage on sperm membrane integrity and other functional characteristics of canine spermatozoa : in vitro bioassay for canine semen Kumi-Diaka J;Badtram G https://doi.org/10.1016/0093-691X(94)90187-N
  13. Vet Clin No Anim Prac v.21 Achieving canine pregnancy by using frozen of chilled extended semen Linde-Forsberg C https://doi.org/10.1016/S0195-5616(91)50054-1
  14. J Repord Fert Fertility in dogs in relation to semen quality and the time and site of insemination with fresh and frozen semen Linde-Forsberg C;Forsberg M
  15. J Reprod Fert Results of 527 controlled artificial inseminations in dogs Linde-Forsberg C;Forsberg M
  16. Theriogenology v.52 Fertility data from 327 artificial inseminationas with frozen-thawed dog semen comparing vaginal and intrauterine deposition: a retrospective study Linde-Forsberg C;Strom Holst B;Govette G https://doi.org/10.1016/S0093-691X(99)00106-5
  17. Theriogenology v.31 Influence of extender, cryopereservative and seminal processing procedures on post thaw motility of canine spermatozoa frozen in staws Olar TT;Bowen RA;Rickett BW https://doi.org/10.1016/0093-691X(89)90550-5
  18. Reprod Dom Anim v.33 Effects of sodium dodecyl sulphate on post thaw dog semen quality during in vitro incubation at 39℃ and 22℃ Pena AI;Barrio F;Quintela LA;Herradon PG https://doi.org/10.1111/j.1439-0531.1998.tb01377.x
  19. Theriogenology v.54 Effects of Equex, one of two step dilution, and two freezing and thawing rates on post-thaw survival of dog spermatozoa Pena AI;Linde-Forsberg C https://doi.org/10.1016/S0093-691X(00)00397-6
  20. J Anim Sci v.47 Effect of Orvus ES Paste on acrosome morphology, motility and fertilizing capacity of frozen-thawed boar sperm Pursel VG;Schulmal LL;Johnson LA
  21. Theriogenology v.47 Effects of Equex-STM paste on viability of frozen-thawed dog spermatozoa during in vitro incubation at 38℃ Rota A;Strom B;Linde-Forsberg C;Rodriguez-Martinez https://doi.org/10.1016/S0093-691X(97)00066-6
  22. Theriogenology v.44 Effects of seminal plasma and three extenders on canine semen stored at 4℃ Rota A;Strom B;Linde-Forsberg C https://doi.org/10.1016/0093-691X(95)00278-G
  23. Reprod Dom Anim v.33 Cryosurvival of dog spermatozoa at different glycerol concentrations and freezing/thawing rates Rota A;Linde-Forsberg C;Vannozzi J;Romagnoli S;Rodriguez-Martinez H https://doi.org/10.1111/j.1439-0531.1998.tb01370.x
  24. Vet Rec v.92 Progress on the use of frozen semen in the dog Seager SWJ;Fletcher WS https://doi.org/10.1136/vr.92.1.6
  25. AI Digest v.17 Successful pregnancies utilizing frozen dog semen Seager SWJ
  26. PhD Thesis University of Minnesota Cryopreservation of canine semen, technique and performance Smith FO
  27. Proc Ann Mtg Soc Theriogenology Update on freezing canine semen Smith FO
  28. Proc 12th Int Congr Anim Reprod(ICAR) v.4 Addition of sodium dodecyl sulphate to the TRIS-citrate extender improves motility and longevity of frozen-thawed canine spermatozoa Thomase PGA;Surman V;Myer-Wallen VN;Concannon PW;Ball BA
  29. Reprod Fertil Dev v.7 Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing funciton Watson PF https://doi.org/10.1071/RD9950871
  30. 대한 수의학회지 v.34 no.4 개에서 동결정액을 이용한 인공수정-Methanol을 이용한 간이 동결방법 김용준;박영재;김병진;유일정
  31. 한국임상수의학회지 v.11 no.2 Methanol을 이용한 개 정액 동결의 융해후 양혼한 활력 및 생존력을 나타내는 정액 처리 조건 김용준;박영재;김병진;유일정
  32. 한국가축위생학회지 v.19 no.1 개 정액 동결시 seeding 처리가 융해 후 정자의 활력 및 생전율에 미치는 효과 김종호;이필돈;유일정;김용준
  33. 한국임상수의학회지 v.8 개 정액의 융해 후 정자의 생존율 향상을 위한 동결방법과 생존율 및 수정능획득 판정에 관한 연구 지동범;김용준