Response of buffalo spermatozoa to low temperatures during cryopreservation
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Abstract
This is the first detailed report on the response of buffalo spermatozoa to low temperatures during freezing. The study determined the critical temperature zone for buffalo spermatozoa and developed a suitable freezing rate for this species. Semen from four Nili-Ravi buffalo bulls diluted in Tris-citric acid was frozen in a programmable freezer. Motion characteristics, plasma membrane integrity and acrosome morphology were determined at +4, 0, –5, –10, –20, –30, –40, –50, –80 and –196°C by removing semen straws from the freezer at exactly these temperatures and rewarming them at 37°C. The first statistical decline in sperm motility and lateral head displacement was observed at –40°C. For all other parameters, there was biphasic decline: for curvilinear velocity, at 0°C and –50°C; and for plasma membrane integrity and acrosome morphology, at –30°C and –50°C. In a second series of experiments, buffalo spermatozoa were frozen using slow (–10°C min–1), medium (–20°C min–1) or fast (–30°C min–1) freezing rates, between –10°C and –80°C. Freezing of buffalo spermatozoa at a rate of –30°C min–1 yielded higher post-thaw motion characteristics, plasma membrane integrity and normal acrosomes. In conclusion, different sperm characteristics respond differently at low temperatures and the freezing of buffalo spermatozoa at a higher rate ensures higher post-thaw semen quality.
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