Journal of Veterinary Clinics (한국임상수의학회지)

The Korean Society of Veterinary Clinics (한국임상수의학회)
권호 표지

Effects of Seeding during Freezing Procedure on Post-Thaw Viability and Acrosome Integrity of Boar Spermatozoa

돼지정액 동결중 식빙처리가 융해후 정자생존율 및 침체형태에 미치는 영향

  • Published : 2004.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

To investigate the effects of seeding during freezing procedure on post-thaw viability, motility, and acrosome integrity of boar spermatozoa, semen from 5 Yorkshire boars were collected for this experiment. Raw semen were diluted with Merck I, subsequently added with cooling diluent containing lactose and egg yolk and with freezing diluent containing glycerol. The diluted semen were frozen on the rack in the styrofoam box filled with liquid nitrogen at the distance of 5 cm or I cm above LN2 level. Seeding was performed to only a group of straws frozen at 5 cm away on the surface of LN2. The frozen semen were thawed in $50^{\circ}C$C water and the viability and local motility were analyzed by sperm analysis imaging system. A part of thawed semen was taken for the examination of morphology of apical ridge of the acrosome to compare with the effect of seeding between the seeding-treated and non treated groups. I. Post-thaw viability was considerably higher in seeding-treated sperm than non-seeding group (p<0.01), however, no difference of local motility was obtained among the groups. 2. At three hours after thawing, viability was also higher in seeding-treated group than non-treated group (p<0.05), along with no difference of motility among the groups. 3. Higher normal acrosome integrity was obtained in the seeding-treated sperm than non-treated groups (p<0.01). 4. Between non-seeded groups, higher normal acrosome integrity was obtained in the sperm group frozen at 5cm upper on the surface of LN2 than that frozen at 1cm away (p<0.01). These results indicated that seeding treatment during freezing boar spermatozoa was beneficial to post-thaw viability and normal acrosome integrity.

Keywords

References

  1. Almid T, Johnson LA. Effects of glycerol concentration, equilibration time and temperature of glycerol addition on post-thaw viability of boar spermatozoa frozen in straws. J Anim Sci 1988; 66: 2899-2905
  2. Andersen K. Morphological abnormalities in the acrosome and nucleus of boar spermatozoa. Nord Vet-Med 1974; 26: 215-218
  3. Bane A. Acrosomal abnormality associated with sterility in boar. Proc IVth Int Congr Anim Reprod., The Hague 1961; 4: 810-817
  4. Bwanga Co, De Braganca M, Einarsson S, et al. Freezing of boar semen in mini and maxi-straws. J Vet Med., A 1990; 37: 651-658 https://doi.org/10.1111/j.1439-0442.1990.tb00958.x
  5. Chen Y, Foote RH, Tobback C, et al. Survival of bull spermatozoa seeded and frozen at different rates in egg yolktris milk extenders. J Dairy Sci 1993; 76: 1028-1034 https://doi.org/10.3168/jds.S0022-0302(93)77430-5
  6. Chiravuch S, Bruns ES, Prins GS. Improvement of post-thaw sperm motility in poor quality human semen. Fertil & Steril 1993; 60(4): 706-710
  7. Coulter GH. Bovine spermatozoa in vitro: a review of storage, fertility estimation and manipulation. Theriogenology 1992; 38: 197-207 https://doi.org/10.1016/0093-691X(92)90230-O
  8. Crister JK, Huse-Benda AR, Aaker DV, et al. Cryopreservation of human spermatozoa I. Effects of holding procedure and seeding on motility, fertilizability, and acrosome reaction. Feril & Steril 1987; 47(4): 656-663
  9. Farrand GD, Elsden RP, Seidel GE. Effect of slow cooling end point temperature on survival of frozen bovine embryos. J Anim Sci 1985; 61(2): 460-465
  10. Fiser PS. Interactions of cooling velocity, warming velocity and glycerol concentration on the survival of frozen-thawed boar sperm. Reprod Dom Anim Suppl 1991; 1: 123-137
  11. Graham EF, Crabo BG, Pace MM. Current status of semen preservation in the ram, boar and stallion. XII Biennial Symposium on Animal Reproduction. J Anim Sci 1978; 47(Suppl. II): 80-119
  12. Hillman KH and Treu H. Deep freezing of boar semen: Demonstration and estimation of morphological and enzyme changes of deep-frozen boar semen. Zuchthygiene 1973; 8:105-112 https://doi.org/10.1111/j.1439-0531.1973.tb00245.x
  13. Hofmo PO and Almid T. Recent developments in freezing of boar semen with special emphasis on cryoprotectants. Reprod Dom Animsuppl 1991; 1: 111-122
  14. Johnson LA. 1980. Artificial insemination of swine: Fertility with frozen boar semen, Copenhagen: Int Pig Vet Congr. 1980: 37
  15. Kasai M, Niwa K, Iritani A. Effects of various cryoprotective agents on the survival of unfrozen and frozen mouse embryos. J Reprod Fert 1981; 63: 175-180 https://doi.org/10.1530/jrf.0.0630175
  16. Kim HK, Ko MS, Kim IC, et al. Deep freezing of boar semen in liquid nitrogen container. Korean J Anim Sci 1989; 31(3): 155-157
  17. Kim JK, Kim CK, Kang MJ, et al. Studies on simplified procedures for freezing and thawing of bovine embryos IV. Effects of simplified procedures of freezing and seeding using a cryoprotectant containing sucrose on the rabbit embryo survival rate determined with the FAD test. Korean J Anim Sci 1988; 31(10): 183-189
  18. Kojima T, Soma T, Oguri N. Effect of ice nucleation by droplet of immobilized silver iodide on freezing of rabbit and bovine embryos. Theriogenology 1988; 30(6): 1199-1207 https://doi.org/10.1016/0093-691X(88)90296-8
  19. Kojima T. Studies on cryopreservation of mammalian embryos induction of ice formation with droplets of silver iodide immobilized in alginate gel. Jpn J Anim Reprod 1991; 37(5): 13-23 https://doi.org/10.1262/jrd1977.37.13P
  20. Kuzan FB, Quinn P. Cryopreservation of mammalimalian embryos. In vitro fertilization and embryo transfer. A Manual of Basic Techniques. 1988: 301-347
  21. Kuzan FB and Quinn P. Cryopreservation of mammalian embryos, In: In vitro fertilization and embryo transfer. NewYork: Plenum Press. 1988: 301-306
  22. Larsson K. Current research on the deep freezing of boar semen. World Rev Anim Prod 1978; XIV: 59-64
  23. Larsson K. Boar sperm viability after freezing and thawing. 1st Intl Conf. Deep freezing boar semen. Uppsala. 1985: 177-188
  24. Leibo SP. The theory and practice of seeding in embryo cryopreservation. Embryo Transfer Newslett. 8(1): 7
  25. Mazur P. Basic concepts in freezing cells. 1st Intl Conf. Deep freezing boar semen. Uppsala. 1985: 91-112
  26. Park Cs, Yang MH, Hwang Ds, et al. Study on fresh and deep-fresh and deep-frozen storage of korean native goat spermatozoa. Korean J Anim Sci 1989; 31(7): 412-419
  27. Parkinson TJ, Whitfield CH. Optimization of freezing conditions for bovine spermatozoa. Theriogenology 1987; 27(5): 781-789 https://doi.org/10.1016/0093-691X(87)90300-1
  28. Parquignon M, Martinat-Botte F, Bariteau F, et al. Preoccupations et connaissances techniques en matiere de reproduction porcine. Jour Rech Porcine en France. 1978: 63-92
  29. Parquignon M. Freezing and thawing extenders for boar spermatozoa. 1st Intl Conf. Deep freezing boar semen, Uppsala. 1985: 129-146
  30. Pursel VG, Johnson LA. Procedure for the preservation of boar spermatozoa by freezing. U.S. Dept Agric ARS. 1971: 44-227
  31. Pursel VG. Advances in preservation of seine spermatozoa. In: H.W. Hawk ed. Beltsville Symponia III. Animal Reproduction Allanheld, Osmun and Co., Montclair, NJ. 1979: 145-15
  32. Rodriguez-Martinez H, Ericksson B, and Lundeheim N. Reproduction in domestic animals: Boar semen preservation. Procd. of the Third international conference on boar semen preservation. Mariensee. 1996: 161-168
  33. Romeny E, Hillman KH, and Richter L. Deep freezing of boar semen 4th communication: Laboratory experiments and inseminations with frozen boar semen using the extender. Hulsenberg IV. Dtsch. tierarztl Wschr. 1974; 81: 353-354
  34. Waberski D, Meding S, Dirksen G, et al. Fertility of long-term stored boar semen: Influence of extender(Androhep and Kiev), storage time and plasma droplets in the semen. Animal Reproduction Science 1994; 36: 145-151 https://doi.org/10.1016/0378-4320(94)90061-2
  35. Westerdorf D, Richter L, and Treu H. On the deep freezing of boar semen: Laboratory and insemination results with the Hulsenbergstraw procedure. Dtsch tierarztl Wschr 1975; 82: 261-267
  36. Whittingham DG. The freezing of mammalian embryos(Eds). Ciba Foundation symposium 52, Amsterdam. 1977: 97-124
  37. 김종호, 이필돈, 유일정, 김용준. 견 정액 동결시 seeding 처리가 응해 후 정자의 활력 및 생존율에 미치는 효과. 한국가축 위생학회지 1995; 19(1): 1-12