The Thai Journal of Veterinary Medicine
Article Title
Comparison of different methods for sperm vitality assessment in frozen-thawed Holstein bull semen
Abstract
The present study evaluated the accuracy of four methods, i.e. eosin-nigrosin, trypan blue, hypo-osmotic swelling test (HOST) and Hoechst 33342 (H342)/propidium iodide (PI) for assessing bovine sperm vitality. Frozen-thawed semen (n = 30) of Holstein bulls was layeredon 40%/80% percoll solutions to isolate viable spermatozoa. An aliquot of viable spermatozoa was kept at 37°C (live sample); the rest was submitted to cold shock (dead sample). The two aliquots were mixed in three proportions, corresponding to 0%, 50% and100% of viable cells; the sperm vitality was analyzed. The percentages of viable spermatozoa evaluated with the four methods significantly correlated with the expected vitality (rs= 0.92 –0.94, P<0.001). Comparing methods of evaluation, the sperm vitality assessed by eosin-nigrosin and trypan blue was comparable (P>0.05) when live and dead samples were used. For the live sample, eosin-nigrosin (82.37 ± 1.48%) and trypan blue (81.00 ± 1.67%) yielded significantly greater results than HOST (50.14 ± 1.68%) and H342/PI (56.69 ± 1.76%) (P<0.001). The percentage of viable spermatozoa in the dead sample (13.45 ± 0.86%) was highest when HOST was exploited (P<0.001); still, the sperm vitality acquired by this technique become the lowest when the live sample was evaluated (P<0.001 to P=0.03). In conclusion, while HOST and fluorescence stains H342/PI with the protocol used in this study are not trustworthy methods, eosin-nigrosin and trypan blue are accurate techniques for sperm vitality assessment in frozen-thawed Holstein bull semen.
Publisher
Faculty of Veterinary Science, Chulalongkorn University
First Page
249
Last Page
255
Recommended Citation
Buranaamnuay, Kakanang
(2019)
"Comparison of different methods for sperm vitality assessment in frozen-thawed Holstein bull semen,"
The Thai Journal of Veterinary Medicine: Vol. 49:
Iss.
3, Article 7.
Available at:
https://digital.car.chula.ac.th/tjvm/vol49/iss3/7