Comparative Evaluation of Motility and Kinematics of Fresh Versus Frozen-Thawed Spermatozoa of Cattle and Buffalo Bull by CASA

DOI: 10.18805/ijar.B-3881    | Article Id: B-3881 | Page : 1188-1194
Citation :- Comparative Evaluation of Motility and Kinematics of Fresh Versus Frozen-Thawed Spermatozoa of Cattle and Buffalo Bull by CASA.Indian Journal of Animal Research.2020.(54):1188-1194
P.K. Pathak, A.J. Dhami D.V. Chaudhari and K.K. Hadiya ajdhami@aau.in
Address : Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science and Animal Husbandry, AAU, Anand-388 001, Gujarat, India.
Submitted Date : 13-07-2019
Accepted Date : 17-10-2019


A study was undertaken on semen of three mature bulls each of Gir, Surti and Murrah breed to evaluate the comparative motion characteristics and kinematics of their fresh and frozen-thawed spermatozoa by Biovis CASA. The ejaculates (n= 24/breed) having >75% initial motility were diluted @ 80 million sperm/ml using TFYG extender and were assessed. Amongst motility traits, the total motile, rapid progressive motile and slow progressive motile spermatozoa percentage decreased significantly by 23.08 - 30.09, 43.57 - 55.18, 9.12 - 22.75 %, respectively, (p<0.01), while non-progressive motile sperm (4.78 - 21.48%) and immotile sperm (164.38 - 178.38 %) percentage increased significantly (p<0.01) in frozen-thawed semen compared to that of fresh semen. The post-thaw quality of semen of all three breeds was in acceptable range. The mean values of sperm velocity/kinematic parameters observed in frozen-thawed semen of Gir, Surti and Murrah bulls, based on total motile sperms, viz., average path velocity,  curvilinear velocity, straight line velocity, linearity, straightness, beat-cross frequency, amplitude of lateral head displacement, dancing frequency and dancing mean decreased significantly by 13.70 - 17.79; 9.76 - 12.95; 13.28 - 21.90; 7.28 - 9.68; 4.36 - 7.79; 15.56 - 25.15; 8.78 - 10.50; 6.16 - 18.67 and 12.98 - 15.96 %, respectively, as compared to that of their fresh semen samples. However, wobbling index remained almost same for both fresh and frozen semen. All motility traits differed but none of kinematics/velocity traits differed significantly between breeds/species. The values of all velocity parameters for progressive motile sperms were higher than total motile sperms in all three breeds. The effect of freezing-thawing on velocity and kinematic attributes was much less compared to absolute sperm motility, and both the fresh and frozen-thawed sperms behaved identically with respect to their velocity and kinematics. The rapid progressive motile sperm in both fresh (r=0.41 to 0.92) and frozen-thawed (r=044 to 0.88) semen had significant correlations with most of their velocity traits, and the later were significantly and positively or negatively inter-related among each other in semen of all three breeds. It was therefore concluded that cryopreservation process significantly reduces the motility and kinematics attributes of bovine spermatozoa and, CASA analysis of fresh semen for motility and velocity traits could predict the post-thawed sperm motility and velocity/ kinematics of spermatozoa.


Buffalo bull Cow bull CASA. Fresh semen Frozen-thawed semen Kinematics Sperm motion characteristics


  1. Amann, R.P. and Waberski, D. (2014). Computer-assisted sperm analysis (CASA): capabilities and potential developments. Theriogenology. 81: 5-17.
  2. Anel L., de Paz P., Alvarez M., Chamorro C.A., Boixo J.C., Manso A., Gonzalez M., Kaabi M. and Anel E. (2003). Field and in vitro assay of three methods for freezing ram semen. Theriogenology. 60: 1293-1308.
  3. Anderson, M., Hellman, T., Holmström, B.G. and Jokinen, L. (1992). Computerized and subjective assessments of post-thaw motility of semen from Finnish Ayrshire AI bulls in relation to non-return rates. Acta. Vet. Scand. 33(1): 89-93.
  4. Budworth, P.R., Amann, R.P. and Chapman, P.L. (1988). Relationships between computerized measurements of motion of frozen-    thawed bulls spermatozoa and fertility. J. Androl. 9: 41-54.
  5. Chaudhary, P.J., Dhami, A.J., Chaudhari, D.V. and Parmar, S.C. (2017). Freezability of cattle and buffalo semen and association of fresh and frozen-thawed sperm quality parameters. Intl. J. Curr. Microbiol. and Appl. Sci. 6(12): 1445-1454.
  6. Christensen, P., Boelling, D., Pedersen, K.M., Korsgaard, I.R. and Jensen, J. (2005). Relationship between sperm viability as determined by flow cytometry and nonreturn rate of dairy bulls. J. Androl. 26: 98-106.
  7. Dhami, A.J., Mohan, G. and Sahni, K.L. (1998). Seasonal influence on the quality and freezability of semen of Friesian and Murrah buffalo bulls. Indian J. Anim. Reprod. 19(1): 55-58.
  8. Dhami, A.J., Panchal, M.T. and Kodagali, S.B. (1990). Comparative biochemical and enzymatic study of Jersey, cross bred and Surti buffalo semen with reference to freezability and fertility. Indian J. Anim. Res. 24(1): 1-8.
  9. Ferrell, P.B., Presicce, G.A., Brockett, C.C. and Foote, R.H. (1998). Quantification of bull sperm characteristics measured by computer-    assisted sperm analysis (CASA) and the relationship to fertility. Theriogenology. 49: 871-879.
  10. Hoflack, G., Opsomer, G., Rijsselaere, T., Van Soom, A., Maes, D., Kruif de, A. and Duchateau, L. (2007). Comparison of computer assisted sperm motility analysis parameters in semen from Belgian Blue and Holstein-Friesian bulls. Reprod. Dom. Anim. 42: 153-161.
  11. Johnson, J., Boone, W. and Blackhurst, D. (1996). Manual versus computer-automated semen analysis. Part I. Comparison of counting chambers. Fertil. Steril. 65: 150-155.
  12. Karthikeya, G. (2003). Computer automated motility and morphometric analysis of bull and buck spermatozoa. MVSc Thesis, Tamil Nadu Veterinary and Anim. Sci. Univ., Chennai, India.
  13. Kathiravan, P., Kalatharan, J., Edwin, M.J. and Veerapandian, C. (2008). Computer automated motion analysis of crossbred bull spermatozoa and its relationship with in vitro fertility in zona-free hamster oocytes: Anim. Reprod. Sci. 104: 9-17.
  14. Kumar, S., Kumar, A., Singh, A. K., Honparkhe, M., Singh, P. and Malhotra, P. (2018). Improvement in post-thaw semen quality by minimizing the lipid peroxidation following herbal treatment in sub fertile buffalo bulls. The Pharma Innovation. 7(5, Part D): 240.
  15. Lenz, R.W., Kjelland, M.E., VonderHaar, K., Swannack, T.M. and Moreno, J.F. (2010). A comparison of bovine seminal quality assessments using different viewing chambers with a computer-assisted semen analyzer. J. Anim. Sci., doi:10.2527/jas.2010- 3056.
  16. Mandal, D.K., Nagpaul, P.K. and Gupta, A.K. (2003). Motion characteristics of Murrah buffalo bull spermatozoa in various seasons and its relationship with functional integrity of the plasmallema. Theriogenology. 60: 349-358.
  17. Mortimer, D., Mortimer, S.T., Ombelet, W., Bosmans, E., Vandeput, H., Vereecken, A., Renier, M. and Hoomans, E. (1998). Value and reliability of CASA systems. Eds. Modern ART in the 2000s. London: Parthenon Publishing Group Limited. 73-89.
  18. Patel, J.B. and Dhami, A.J. (2013). Computer assisted sperm analysis of fresh and frozen-thawed HF x Kankrej (F1) bulls’ semen and their interrelationship. Indian J. Anim. Res. 47: 315-320.
  19. Patel, J.B. and Dhami, A.J. (2016). Computer assisted sperm analysis of fresh and frozen-thawed buffalo semen and their interrelationship. Indian J. Anim. Res. 50(1): 8-13.
  20. Patel, J.B., Dhami, A.J., Patel, P.A. and Savalia, F.P. (2012). Assessment of motility and velocity parameters of freshly ejaculated spermatozoa of Bubalus bubalis and Bos indicus bulls through computer assisted semen analyzer. Indian J. Field Vets. 8(1): 18-22.
  21. Patel, N.P., Patel, S.H. and Devanand, C.P. (2010). Comparative studies on the assessment of the bovine sperm motility by conventional method and CASA System (HTM IVOS). Proc. XXVI Annual Convention of ISSAR and International Symposium, GBPUA and T, Pantnagar, 10-12 Nov., p. 11.
  22. Pathak, P.K., Dhami, A.J. and Chaudhari, D.V. (2018). Correlations of fresh sperm quality and kinematic attributes to those of post-    thawed sperm of bulls assessed by CASA. Intl. J. Curr. Microbiol. Appl. Sci. 7(9): 2606-2615.
  23. Perumal, P., Selvaraju, S., Selvakumar, S., Barik, A., Mohanty, D., Das, S., Das, R. and Mishra, P. (2011). Effect of pre-freeze addition of cysteine hydrochloride and reduced glutathione in semen of crossbred Jersey bulls on sperm parameters and conception rates. Reprod. Dom. Anim. 46(4): 636-641.
  24. Ramachandran, N., Verma G.S., Gupta, A.K., Raina, V.S., Mohanty, T.K. and Singh, M.K. (2007). Prediction of fertility based on frozen-thawed seminal traits in Sahiwal bulls. Indian J. Anim. Sci. 77(8): 884-887.
  25. Rasul, Z., Ahmad, N. and Anzar, M. (2001). Changes in motion characteristics, plasma membrane integrity, and acrosome morphology during cryopreservation of buffalo spermatozoa. J. Androl. 22: 278-283.
  26. Rhodes, F., Galina, C. S., Duchateau, A. and Sato, C. (1985). An investigation into the properties of bovine semen in the Mexican tropics. World Review Anim. Prod. 21(2): 15-19.
  27. Rodriguez-Montaña, D. and Roa-Guerrero, E. (2017). Objective assessment of bull sperm motility parameters using computer vision algorithms. African J. Biotech. 16(37): 1871-1881.
  28. Shelke, V.B. and Dhami, A.J. (2001). Comparative evaluation of physico-morphological attributes and freezability of semen of Gir cattle (Bos indicus) and Jafarabadi buffalo (Bubalus bubalis) bulls. Indian J. Anim. Sci. 71: 319-324.
  29. Snedecor, G.W. and Cochran, W.G. (1994). Statistical Methods. 14th edn. Oxford and IBH Publishing House, New Delhi, India.
  30. Verstegen, J., Iguer-Ouada, M. and Onclin, K. (2002). Computer assisted semen analyzers in andrology research and veterinary practice. Theriogenology. 57(1): 149-179.
  31. Viswanath R. and Shannon P. (1997). Do sperm cells age? A review of the physiological change in sperm during storage at ambient temperature. Reprod. Fert. Dev. 9: 321-331. 

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