Effectiveness of Giemsa and Modified Trypan Blue-GiemsaStaining for the Assessment of Acrosome Integrity in Bull andBuck semen

Authors

  • Ravichandran Ruthrakumar Department of Veterinary Gynaecology and Obstetrics, Veterinary College and Research Institute, Namakkal – 637 002 Tamil Nadu Veterinary and Animal Sciences University (TANUVAS), Chennai
  • Murugan Sabarinathan Department of Veterinary Gynaecology and Obstetrics, Veterinary College and Research Institute, Namakkal – 637 002 Tamil Nadu Veterinary and Animal Sciences University (TANUVAS), Chennai
  • Manokaran Dhanush Department of Veterinary Gynaecology and Obstetrics, Veterinary College and Research Institute, Namakkal – 637 002 Tamil Nadu Veterinary and Animal Sciences University (TANUVAS), Chennai
  • Venkatachalam Kalaiyarasan Department of Veterinary Gynaecology and Obstetrics, Veterinary College and Research Institute, Namakkal – 637 002 Tamil Nadu Veterinary and Animal Sciences University (TANUVAS), Chennai
  • Duraisamy Gopikrishnan Department of Veterinary Gynaecology and Obstetrics, Veterinary College and Research Institute, Namakkal – 637 002 Tamil Nadu Veterinary and Animal Sciences University (TANUVAS), Chennai
  • Mahakrishnan Palanisamy Department of Veterinary Gynaecology and Obstetrics, Veterinary College and Research Institute, Namakkal – 637 002 Tamil Nadu Veterinary and Animal Sciences University (TANUVAS), Chennai
  • Mani Selvaraju Department of Veterinary Gynaecology and Obstetrics, Veterinary College and Research Institute, Namakkal – 637 002 Tamil Nadu Veterinary and Animal Sciences University (TANUVAS), Chennai

DOI:

https://doi.org/10.48165/ijar.2024.45.01.12

Keywords:

Acrosomal integrity, Buck, Bull, Giemsa, Trypan blue

Abstract

   

The estimation of acrosomal integrity is crucial for predicting the fertility of spermatozoa. The semen quality analysis  demands assessment techniques to be reliable, quick and easy to perform for routine application in semen station. A study  was conducted to assess the acrosomal integrity of Kangayam cattle and Salem Black goat using Giemsa stain and modified  Trypan Blue-Giemsa stain. For this procedure, 70 numbers of frozen semen straws of Kangayam bull and Salem Black bucks  were stained. The staining quality and acrosomal integrity were assessed at the interval of 3, 5 and 24 hours interval up to 5  days, and evaluated as acrosomal intact and non-intact spermatozoa. In Kangayam bull semen, the quality of the staining  was good and better in trypan blue – Giemsa stain than Giemsa alone but for Salem Black buck semen Giemsa showed  better staining quality than trypan blue – Giemsa combination. However, the working solution of Giemsa once prepared can  be used for 4 days but the trypan blue-Giemsa stain can be used for only 24 hours once prepared. Hence, stain is required  to be prepared on daily basis for routine semen evaluation in semen stations. As a conclusion trypan blue – Giemsa staining  method is recommended for better staining of acrosome in bull semen and Giemsa alone in Buck semen.  

References

Aitken, R. J. (2006). Sperm function tests and fertility. Int. J. Androl.,29(1):69-75.

Chelucci, S., Pasciu, V., Succu, S., Addis, D., Leoni, G. G., Manca, M, E, Naitana, S. and Berlinguer, F. (2015). Soybean leci thin-based extender preserves spermatozoa membrane integrity and fertilizing potential during goat semen cryo preservation. Theriogenology,14:1 -12.

Chowdhury, S., Das, S., Gupta, T., Sana, D. and Bose, S. (2014). Evaluation of frozen semen by acrosomal integrity and sperm concentration- two vital quality parameters of male fertility in bovines. Explor. Anim. Med. Res.,4(1):101-107.

Graham, J. K. and Moce, E. (2005). Fertility evaluation of frozen/ thawed semen. Theriogenology,64(3):492-504.

Huo, L. J., Yue, K. Z. and Yang, Z. M. (2002). Characterization of viability, mitochondrial activity, acrosomal integrity and capacitation status in boar sperm during in vitro stor age at different ambient temperatures. Reprod. Fert. Dev., 14(8):509-514.

Insani, K., Sri, R., Agung, P. and Aries, S. (2014). Kadar MDA spermatozoa setelahpembekuan. Biotropika.,3(2): 11-16.

Jankovičová, J., Michal, S., Jana, A. and Ľubica, H. (2008). Acrosomal and viability status of bovine spermatozoa eval uated by two staining methods. Acta Veterinaria Hungarica, 56(1):133–137.

Kastelic, J. P. and Thudathil, J. C. (2008). Breeding soundness evaluation and semen analysis for predicting bull fertility. Reprod. Dom. Anim.,43:368-373.

Neild, D.N., B. M. Gadella, A. Aguero, T.A.E. Stout and B. Colenbrander. (2005). Capacitation, acrosome function and chromatin structure in stallion sperm. Anim. Reprod. Sci., 89: 47-56.

Ostermeier, G. C., Sargeant, G. A., Yandell, B. S., Evenson, D. P. and Parrish, J. J. (2001). Relationship of bull fertility to sperm nuclear shape. J. Androl.,4:595-603.

Patrat, C., Serres, C. and Jouannet, P. (2000). The acrosome reac tion in human spermatozoa. J, Cell Biol.,92(3-4):255-266. Prihantoko, K.D., Yuliastuti, F., Haniarti, H., Kusumavati, A., Widayati, D.T. and Budiyanto, A. (2020). The acrosome integrity examination of post-thawed spermatozoa of sev eral Ongole grade bull in Indonesia using Giemsa staining method. IOP Conf. Ser. Earth Environ. Sci., 478: 012042. Rath, D., Bathgate, R., Rodriguez-Martinez, H., Roca, J., Strzezek, J. and Waberski, D. (2009). Recent Advances in Boar Semen Cryopreservation. Society of Reprod. Fert. Suppl.,66: 51–66. Talbot, P. and Chacon, R. (1981): A triple stain technique for evaluating normal acrosome reactions of human sperm. J. Exp. Zool.,215:201-208.

Downloads

Published

2024-04-30

Issue

Vol. 45 No. 1 (2024): The Indian Journal of Animal Reproduction( june)

Section

Research Article

How to Cite

Ruthrakumar , R., Sabarinathan, M., Dhanush, M., Kalaiyarasan, V., Gopikrishnan, D., Palanisamy , M., & Selvaraju, M. (2024). Effectiveness of Giemsa and Modified Trypan Blue-GiemsaStaining for the Assessment of Acrosome Integrity in Bull andBuck semen. The Indian Journal of Animal Reproduction, 45(1), 53–57. https://doi.org/10.48165/ijar.2024.45.01.12
Crossref
Scopus
Google Scholar
Europe PMC