Banner
Current IssueEditorial BoardOnline First ArticlesArchive
Current IssueEditorial BoardOnline First ArticlesArchive

volume 45 issue 4 (december 2011) : 247 - 255

EFFECT OF DIFFERENT SEASONS AND THAWING PROTOCOLS ON CERTAIN SEMINAL ATTRIBUTES OF INDIAN STANDARD BRED STALLION SEMEN PRESERVED USING 1% GLYCEROL AND 1% DIMETHYL FORMAMIDE AS CRYOPROTECTANT

D
Devender Kumar
D
Dinesh Jhamb*
A
Atul Saxena
1Equine Fertility Research Center. Equine Breeding Stud, Babugarh, Ghaziabad -245 201, India

  • Submitted|

  • First Online |

  • doi

Cite article:- Kumar Devender, Jhamb* Dinesh, Saxena Atul (2025). EFFECT OF DIFFERENT SEASONS AND THAWING PROTOCOLS ON CERTAIN SEMINAL ATTRIBUTES OF INDIAN STANDARD BRED STALLION SEMEN PRESERVED USING 1% GLYCEROL AND 1% DIMETHYL FORMAMIDE AS CRYOPROTECTANT. Indian Journal of Animal Research. 45(4): 247 - 255. doi: .

ABSTRACT

Standard bred stallion semen was collected during the breeding (n=5) (March-October) and non breeding (n=5) (November-January) seasons using artificial vagina for seven occasions on alternate days from each stallion for both seasons. Semen was evaluated for physical attributes, motility and membrane integrity, diluted using INRA 82 extender to which 2% egg yolk, 1% glycerol and 1% dimethyl formamide filled in french medium straws and cryopreserved by instant freezing over LN2 Vapors. After 24 hours of storage, semen was thawed using three different protocols Treatment 1 (T1) 37ÚC for 30 secs, Treatment 2 (T2) 60ÚC for 8secs and Treatment 3 (T3) 75ÚC for 7 secs. The post thaw motility, post thaw viability and post thaw HOS positive sperms were significantly higher during the non breeding season. The thawing protocol T2 was found to be the best during the breeding as well was non-breeding seasons.

REFERENCES

  1. Bradford, L. L. and Buhr, M. M. (2002). Function of cryopreserved horse semen is improved by optimal thawing rates. Equine Vet. Sci.., 7: 145 – 173.
  2. Cabrita, E., Alveraj, R., Anel, E. and Heeraez, M. P. (1999). The hypo osmotic swelling test performed with coulter counter. A method to assay functional integrity of sperm membrane in rainbow trout. Animal Reproduction Sci.., 55 (3-4): 279- 287.
  3. Cristanelli, M. J., Squires E. L., Amann, R. P. and Pickett, E. W. (1984). Fertility of stallion semen processed frozen and thawed by a new procedure. Theriogenology., 22: 39-45.
  4. Cochran , J. D ., Amann, R. P., Froman, D. P and Pickett, B. W. (1984). Effect of centrifugation, glycerol level, cooling to 5°C, freezing rate and thawing rate on the post thaw motility of equine spermatozoa. Theriogenology., 22: 25-38.
  5. Coiza, de la, Cueva, F.I., Regaud, T. Bonet, S. Miro, J. Eriz, M. Rodriguez-Gil J E. (1997). Subjecting horse spermatozoa to hypo osmotic incubation effects of ouabain. Theriogenology., 47: 765-84.
  6. Davis-Morel, M.C.G.: (1999). Equine Artificial Insemination. CAB International, Wallingford, Oxon, OX10 8DE UK.
  7. Dott, H. M. and Foster, G. C. (1972). A technique for studying the morphology of mammalian spermatozoa which are eosinophilic in differential ‘live/Dead’ stain. J. Reprod and Fertility., 29 : 443-445.
  8. Dell’aqua Jr, J.A., Papa, F. O., Alvarenga, M. A., Zahn, F. S. (2001.) Effects of warming rate on sperm parameters and of insemination site and dose on the fertility of equine frozen semen. 3rd international symposium on stallion reproduction, Colorado State University, Fort Collins, USA. pp 64.
  9. Grondahl, C., Grondahl, M. L., Hyttel, P. and Greve, T. (1994). Acrosomal status in fresh and frozen thawed stallion spermatozoa evaluated by scanning electron microscopy, Anatomica Embryologica.., 190: 195-200.
  10. Graham, J. K. (1996). Methods for induction of capitation and acrosome reaction of stallion spermatozoa. Veterinary Clinics of North America Equine Practice., 12(1): 131-147.
  11. Henry, M., Snoeck, P.P.N. and Cottorello, A.C.P. (2002). Post thaw spermatozoa plasma membrane integrity and motility of stallion semen frozen with different cryoprotectants. Theriogenology. 58: 245-248.
  12. Johnson, L. (1991). Seasonal differences in equine spermatocytogenesis. Biology of. Reprod.., 44: 284-291.
  13. Jeyendran, R. S., VanderVen, H .H., Perez-Pelaez, M., Grabo, B. G., Zaneveld, L. J. D. (1984). Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J. Reprod. Fertil., 70: 219-225.
  14. Loomis, P. R. and Squires, E. L. (2005). Frozen semen management in equine breeding programs. Theriogenology., 64: 480-49.
  15. Loomis, P. R., Amann, R. P., Squires, E. L. and Pickett, B. W. (1983). Fertility of unfrozen and frozen stallion spermatozoa extended in EDTA-lactose-egg yolk and packaged in straws. J. Anim. Sci., 56: 687-693.
  16. Mazur, P. (1984) Freezing of living cells: mechanisms and implications. Amer. J. Physiol.., 247: 125-142.
  17. Magistrini, M. Vidament, C., Labbe, A., Gennevieve, Stradaioli, G. (2001). Assessment of Sperm Quality. In proceedings of 3rd international symposium on stallion reproduction, Colorado State University, Fort Collins, USA. 24.
  18. Miragaya, M. H., Chaves, M. G., Neild, D. M., Berretta, C. and Aguero, A. (2001). Artificial Insemination using stallion semen cryopreserved with a simple manual method. In Proceedings of 3rd International Symposium on Stallion Reproduction, Colorado State University, Fort Collins, USA.pp 56.
  19. Metcalf, E. S. (2005). Optimizing pregnancy rates using frozen-thawed equine semen. (In Proceedings of 4th international symposium on Stallion Reproduction) Colorado State University, Fort Collins, USA. Anim. Reprod. Sci.,. 89: 297.
  20. Neild, D. M., Chaves, M. G, Flores M. Miragaya, M. H. Gonzalez, E., Aguero, A. (2000.) The HOS test and its relationship to fertility in the stallion. Andrologia.., 21: 351-355.
  21. Nie, G. J. and Wenzel, J.G. W. (2001). Adaptation of the hypo osmotic swell test to assess functional integrity of stallion spermatozoal plasma membranes. Theriogenology. 55: 1005-1018.
  22. Pant, H.C., Mittal, A. K., Patel, S. H., Shukla, H. R., Kasiraj, R., Prabhker, J. H. (2002). The hypo- osmotic swelling test. an assay of cell membrane integrity and quality of frozen thawed buffalo semen. Indian J. Anim. Reprod.. 23 (1): 8-11.
  23. Papa, F. O., Alvarenga, M. A., Sousa, D. B., Groh, T. M., Carvalho, L. M., Melo, D. S., Leme, D. P., Ferreira, J. C. P., Dell’ Aqua Jr., J. A. (2001) Effect of extender osmolarity and cooling to 50C on motility and membrane integrity of equine frozen spermatozoa. In Proceedings of 3rd International Symposium on Stallion Reproduction, Colorado State University, Fort Collins, USA. pp 63.
  24. Samper, J. C., Hellander, J. C. and Carbo, B. C. (1991). Relationship between the fertility of fresh and frozen stallion semen and semen quality. J. Reprod.. Fertilit., Supplement. 44:107-114
  25. Snedecor, G. W. and Cochran, W. G. (1971). Statistical Methods. 6th edn. The Iowa State Univ. Press. USA.
  26. Squires, E.L., Keith, S.L. and Graham, J.K. (2004). Evaluation of alternative cryoprotectants for preserving stallion spermatozoa. Theriogenology. 62: 1056-1065.
  27. . Zhang, J. J., Boyle, M. S., Smith, C. A. and Moore, H. D. M. (1990). Acrosome reaction of stallion spermatozoa evaluated with monoclonal antibody and zona-free hamster eggs. Molecular Reprod. Develop.. 27: 152-158.
Published In
Indian Journal of Animal Research
Article Metrics
Metrics Icon
0
Views
0
Citations
Reviewed By
Share Article
Download Article
In this Article
    Contact us
    Follow us

    volume 45 issue 4 (december 2011) : 247 - 255

    EFFECT OF DIFFERENT SEASONS AND THAWING PROTOCOLS ON CERTAIN SEMINAL ATTRIBUTES OF INDIAN STANDARD BRED STALLION SEMEN PRESERVED USING 1% GLYCEROL AND 1% DIMETHYL FORMAMIDE AS CRYOPROTECTANT

    D
    Devender Kumar
    D
    Dinesh Jhamb*
    A
    Atul Saxena
    1Equine Fertility Research Center. Equine Breeding Stud, Babugarh, Ghaziabad -245 201, India

    • Submitted|

    • First Online |

    • doi

    Cite article:- Kumar Devender, Jhamb* Dinesh, Saxena Atul (2025). EFFECT OF DIFFERENT SEASONS AND THAWING PROTOCOLS ON CERTAIN SEMINAL ATTRIBUTES OF INDIAN STANDARD BRED STALLION SEMEN PRESERVED USING 1% GLYCEROL AND 1% DIMETHYL FORMAMIDE AS CRYOPROTECTANT. Indian Journal of Animal Research. 45(4): 247 - 255. doi: .

    ABSTRACT

    Standard bred stallion semen was collected during the breeding (n=5) (March-October) and non breeding (n=5) (November-January) seasons using artificial vagina for seven occasions on alternate days from each stallion for both seasons. Semen was evaluated for physical attributes, motility and membrane integrity, diluted using INRA 82 extender to which 2% egg yolk, 1% glycerol and 1% dimethyl formamide filled in french medium straws and cryopreserved by instant freezing over LN2 Vapors. After 24 hours of storage, semen was thawed using three different protocols Treatment 1 (T1) 37ÚC for 30 secs, Treatment 2 (T2) 60ÚC for 8secs and Treatment 3 (T3) 75ÚC for 7 secs. The post thaw motility, post thaw viability and post thaw HOS positive sperms were significantly higher during the non breeding season. The thawing protocol T2 was found to be the best during the breeding as well was non-breeding seasons.

    REFERENCES

    1. Bradford, L. L. and Buhr, M. M. (2002). Function of cryopreserved horse semen is improved by optimal thawing rates. Equine Vet. Sci.., 7: 145 – 173.
    2. Cabrita, E., Alveraj, R., Anel, E. and Heeraez, M. P. (1999). The hypo osmotic swelling test performed with coulter counter. A method to assay functional integrity of sperm membrane in rainbow trout. Animal Reproduction Sci.., 55 (3-4): 279- 287.
    3. Cristanelli, M. J., Squires E. L., Amann, R. P. and Pickett, E. W. (1984). Fertility of stallion semen processed frozen and thawed by a new procedure. Theriogenology., 22: 39-45.
    4. Cochran , J. D ., Amann, R. P., Froman, D. P and Pickett, B. W. (1984). Effect of centrifugation, glycerol level, cooling to 5°C, freezing rate and thawing rate on the post thaw motility of equine spermatozoa. Theriogenology., 22: 25-38.
    5. Coiza, de la, Cueva, F.I., Regaud, T. Bonet, S. Miro, J. Eriz, M. Rodriguez-Gil J E. (1997). Subjecting horse spermatozoa to hypo osmotic incubation effects of ouabain. Theriogenology., 47: 765-84.
    6. Davis-Morel, M.C.G.: (1999). Equine Artificial Insemination. CAB International, Wallingford, Oxon, OX10 8DE UK.
    7. Dott, H. M. and Foster, G. C. (1972). A technique for studying the morphology of mammalian spermatozoa which are eosinophilic in differential ‘live/Dead’ stain. J. Reprod and Fertility., 29 : 443-445.
    8. Dell’aqua Jr, J.A., Papa, F. O., Alvarenga, M. A., Zahn, F. S. (2001.) Effects of warming rate on sperm parameters and of insemination site and dose on the fertility of equine frozen semen. 3rd international symposium on stallion reproduction, Colorado State University, Fort Collins, USA. pp 64.
    9. Grondahl, C., Grondahl, M. L., Hyttel, P. and Greve, T. (1994). Acrosomal status in fresh and frozen thawed stallion spermatozoa evaluated by scanning electron microscopy, Anatomica Embryologica.., 190: 195-200.
    10. Graham, J. K. (1996). Methods for induction of capitation and acrosome reaction of stallion spermatozoa. Veterinary Clinics of North America Equine Practice., 12(1): 131-147.
    11. Henry, M., Snoeck, P.P.N. and Cottorello, A.C.P. (2002). Post thaw spermatozoa plasma membrane integrity and motility of stallion semen frozen with different cryoprotectants. Theriogenology. 58: 245-248.
    12. Johnson, L. (1991). Seasonal differences in equine spermatocytogenesis. Biology of. Reprod.., 44: 284-291.
    13. Jeyendran, R. S., VanderVen, H .H., Perez-Pelaez, M., Grabo, B. G., Zaneveld, L. J. D. (1984). Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J. Reprod. Fertil., 70: 219-225.
    14. Loomis, P. R. and Squires, E. L. (2005). Frozen semen management in equine breeding programs. Theriogenology., 64: 480-49.
    15. Loomis, P. R., Amann, R. P., Squires, E. L. and Pickett, B. W. (1983). Fertility of unfrozen and frozen stallion spermatozoa extended in EDTA-lactose-egg yolk and packaged in straws. J. Anim. Sci., 56: 687-693.
    16. Mazur, P. (1984) Freezing of living cells: mechanisms and implications. Amer. J. Physiol.., 247: 125-142.
    17. Magistrini, M. Vidament, C., Labbe, A., Gennevieve, Stradaioli, G. (2001). Assessment of Sperm Quality. In proceedings of 3rd international symposium on stallion reproduction, Colorado State University, Fort Collins, USA. 24.
    18. Miragaya, M. H., Chaves, M. G., Neild, D. M., Berretta, C. and Aguero, A. (2001). Artificial Insemination using stallion semen cryopreserved with a simple manual method. In Proceedings of 3rd International Symposium on Stallion Reproduction, Colorado State University, Fort Collins, USA.pp 56.
    19. Metcalf, E. S. (2005). Optimizing pregnancy rates using frozen-thawed equine semen. (In Proceedings of 4th international symposium on Stallion Reproduction) Colorado State University, Fort Collins, USA. Anim. Reprod. Sci.,. 89: 297.
    20. Neild, D. M., Chaves, M. G, Flores M. Miragaya, M. H. Gonzalez, E., Aguero, A. (2000.) The HOS test and its relationship to fertility in the stallion. Andrologia.., 21: 351-355.
    21. Nie, G. J. and Wenzel, J.G. W. (2001). Adaptation of the hypo osmotic swell test to assess functional integrity of stallion spermatozoal plasma membranes. Theriogenology. 55: 1005-1018.
    22. Pant, H.C., Mittal, A. K., Patel, S. H., Shukla, H. R., Kasiraj, R., Prabhker, J. H. (2002). The hypo- osmotic swelling test. an assay of cell membrane integrity and quality of frozen thawed buffalo semen. Indian J. Anim. Reprod.. 23 (1): 8-11.
    23. Papa, F. O., Alvarenga, M. A., Sousa, D. B., Groh, T. M., Carvalho, L. M., Melo, D. S., Leme, D. P., Ferreira, J. C. P., Dell’ Aqua Jr., J. A. (2001) Effect of extender osmolarity and cooling to 50C on motility and membrane integrity of equine frozen spermatozoa. In Proceedings of 3rd International Symposium on Stallion Reproduction, Colorado State University, Fort Collins, USA. pp 63.
    24. Samper, J. C., Hellander, J. C. and Carbo, B. C. (1991). Relationship between the fertility of fresh and frozen stallion semen and semen quality. J. Reprod.. Fertilit., Supplement. 44:107-114
    25. Snedecor, G. W. and Cochran, W. G. (1971). Statistical Methods. 6th edn. The Iowa State Univ. Press. USA.
    26. Squires, E.L., Keith, S.L. and Graham, J.K. (2004). Evaluation of alternative cryoprotectants for preserving stallion spermatozoa. Theriogenology. 62: 1056-1065.
    27. . Zhang, J. J., Boyle, M. S., Smith, C. A. and Moore, H. D. M. (1990). Acrosome reaction of stallion spermatozoa evaluated with monoclonal antibody and zona-free hamster eggs. Molecular Reprod. Develop.. 27: 152-158.
    In this Article
      Contact us
      Follow us
      Published In
      Indian Journal of Animal Research

      Editorial Board

      View all (0)