Evaluation of Spermatological Parameters of Frozen Bull Semen with Conventional and CASA (Computer-Aided Sperm Analysis) Method

Abstract

The aim of this study was to compare the assessments made by the CASA system and subjective method (by using phase-contrast microscope) for spermatological examinations of imported and locally produced semen. Frozen semen (imported and local production) belonging to 20 different bulls was examined by phase contrast light microscope method (conventional method) and method supported by computer (CASA method) for evaluating the principle semen characteristics. For imported semen, considering the average values of samples examined by the two methods, significant differences (P<0.05) were observed in the values of motility and concentration as well as the rates of abnormal and dead spermatozoa. For domestic semen, significant differences (P<0.05) were observed for the concentration of samples by the conventional evaluations and for the assessment of motility and concentration by the CASA method. It was observed that there were significantly (P<0.05) higher data only for the concentration assessed by both methods, while no such differences between the values of motility as well as the rates of abnormal and dead spermatozoa were found when considering the general average rates. By using the two methods, findings from the examination of pre-determined parameters were compared and their reliabilities were displayed herein.

Keywords

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Dondurulmuş Boğa Spermalarında Spermatolojik Parametrelerin Konvansiyonel ve CASA (Bilgisayar Destekli Sperm Analiz) Yöntemiyle Değerlendirilmesi

Abstract

Bu araştırmada, ithal ve yerli üretim spermaların ülkemizde geleneksel yöntemlerle faz-kontrast ışık mikroskobu kullanılarak subjektif olarak yapılan spermatolojik muayenelerinin, CASA sistemiyle yapılan değerlendirmelerle karşılaştırılması amaçlandı. Farklı 20 boğaya ait dondurulmuş spermalar (ithal ve yerli üretim) faz-kontrast ışık mikroskobu yöntemi (konvansiyonel yöntem) ve bilgisayar destekli sperm analiz yöntemi (CASA yöntemi) ile muayene edildi ve başlıca spermatolojik özellikler yönüyle değerlendirildi. İthal spermalarda muayene edilen örneklerin ortalama değerleri arasında spermatozoa motilitesi, spermatozoa yoğunluğu, anormal spermatozoa ve ölü spermatozoa oranları arasında gözlenen farklılıklar istatistiksel açıdan önemli oldu (P<0,05). Yerli spermalarda ise konvansiyonel yöntemle numuneler arasında saptanan spermatozoa yoğunluğu yönüyle gözlenen farklılıklar ve CASA yöntemiyle elde edilen spermatozoa motilitesi ve spermatozoa yoğunluğu bakımından gözlenen farklılıklar istatistiksel açıdan önemli bulundu (P<0,05). Genel ortalama değerlere bakıldığında konvansiyonel ve CASA yöntemiyle elde edilen sonuçlara göre yalnızca spermatozoa yoğunluğunda konvansiyonel yöntemle önemli ölçüde daha yüksek veriler alındığı, spermatozoa motilitesi, anormal spermatozoa oranı ve ölü spermatozoa değerlerindeki farklılıkların istatistiksel açıdan önemsiz olduğu belirlendi (P<0,05). Her iki yöntemle, belirlenen muayene parametrelerine ilişkin bulgular karşılaştırıldı ve yöntemlerin güvenilirlikleri ortaya konuldu.

Keywords

• Boğa, CASA, dondurulmuş sperma, konvansiyonel muayene, spermanın değerlendirilmesi

References

1. Budworth, P.R., Amann, R.P., Chapman, P.L., 1988. Relationships between computerized measurements of motion of frozen-thawed bull spermatozoa and fertility. Journal of Andrology 9, 41-54.
2. Davis, I.S., Bratton, R.W., Foote, R.H., 1963. Livability of bovine spermatozoa at 5°C in Tris-buffered and citratebuffered yolk-glycerol extenders. Journal of Dairy Science 46, 57-60.
3. Davis, R.O., Katz, D.F., 1992. Standardization and comparability of CASA instruments. Journal of Andrology 13, 81-86.
4. Davis, R.O., Katz, D.F., 1993. Operational standards for CASA instruments. Journal of Andrology 14, 385-396.
5. Defoin, L., Granados, A., Donnay, I., 2008. Analysing motility parameters on fresh bull semen could help to predict resistance to freezing: A preliminary study. Reproduction in Domestic Animals 43, 606-611.
6. Didion, B.A., 2008. Computer-assisted semen analysis and its utility for profiling boar semen samples. Theriogenology 70, 1374-1376.
7. Farrell, P.B., Presicce, G.A., Brockett, C.C., 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.
8. Gıda Tarım ve Hayvancılık Bakanlığı, 2014. HAYGEM kayıtları: http://www.tarim.gov.tr/sgb/Belgeler/SagMenuVeriler/HAYGEM.pdf.

9. Gravance, C.G., Garner, D.L., Pitt, C., Vishwanath, R., SaxGravanca, S.K., Casey, P.J., 1999. Replicate and technician variation associated with computer aided bull sperm head morphometry analysis (ASMA). International Journal of Andrology 22, 77-82.
10. Hoflack, G., Opsomer, G., Rijsselaere, T., Van Soom, A., Maes, D., de Kruif, A., Duchateau, L., 2007. Comparison of computer-assisted sperm motility analysis parameters in semen from Belgian blue and Holstein-Friesian bulls. Reproduction in Domestic Animal 42, 153-161
11. Januskauskas, A., Zilinskas, H., 2002. Bull semen evaluation post-thaw and relation of semen characteristics to bull’s fertility. Veterinarija ır Zootechnika 17, 26-39. Kastelic, J.P., Thundathil, J.C., 2008. Breeding soundness evaluation and semen analysis for predicting bull fertility. Reproduction in Domestic Animals 43, 368-373.
12. Rodriguez Martinez, H., 2000. Evaluation of Frozen Semen: Traditional and New Approaches. In: Chenoweth, P.J. (Ed.), Topics in Bull Fertility. International Veterinary Information Service, Ithaca, www.ivis.org, (Accessed: 24 June, 2012).
13. Sundararaman, M.N., Kalatharan, J., Thilak Pon Jawahar, K., 2007. Analyses of morphological and morphometrical deviations of bull spermatozoa by Computer Assisted Semen Analysis technique. Asian Journal of Animal and Veterinary Advances 2, 196-204.
14. Tardiff, A.L., Farrell, P.B., Trouern-Trend, V., Foote, R.H., 1997. Computer- Assisted Sperm Analysis for assessing ınitial semen quality and changes during storage at 5°C. Journal of Dairy Science 80, 1606-1612.
15. Tekin, N., 1994. Spermanın muayenesi ve değerlendirilmesi. Alaçam, E. (Ed): Evcil Hayvanlarda Reprodüksiyon, Suni Tohumlama, Doğum ve İnfertilite. Dizgievi, Konya. pp.69-79.
16. Uysal, O., Bucak, M.N., Yavaş, İ., Varışlı, Ö., 2007. Effect of various antioxidants on the quality of frozen-thawed bull semen. Journal of Animal and Veterinary Advances 6, 1362-1366.
17. Uysal, O., Korkmaz, T., Bucak, M.N., Yavaş, İ., 2006. Evaluation by hypoosmotic swelling-eosine test of cryopreserved bovine spermatozoa, Indian Veterinary Journal 83, 557-559.
18. Verstegen, J., Iguer-Ouada, M., Onclin, K., 2002. Computer assisted semen analyzers in andrology research and veterinary practice. Theriogenology 57, 149-179.