Programmable fast-freezing method improves the post-thaw motion dynamics, integrities of plasmalemma, mitochondrial transmembrane, DNA and, acrosome, and in vivo fertility of water buffalo (Bubalus bubalis) spermatozoa.
The effects of freezing methods (FR1, nonprogrammable/static, 5 cm above liquid nitrogen [LN2] for 10 min, plunging in LN2; FR2, programmable medium, +4°C to -15°C at 3°C min-1, from -15 to -80°C at 10°C min-1and final holding for 1 min at -80°C, plunging in LN2; FR3, programmable fast, from initial holding at +4°C for 2 min, from +4°C to -20°C at 10°C min-1, from -20°C to -100°C at 30°C min-1, final holding for 1 min at -100°C and plunging in LN2) were assessed on post-thaw in vitro quality and in vivo fertility of water buffalo spermatozoa. Mean sperm progressive motility (%), rapid velocity (%), average path velocity (μm s-1), straight line velocity (μm s-1), curved line velocity (μm s-1), integrities (%) of plasmalemma, mitochondrial transmembrane, DNA and acrosome were higher (p < .05) in samples cryopreserved with FR3 compared to FR1 and FR2. Similarly, in vivo fertility (%) of buffalo spermatozoa was higher (p < .05) with FR3 than FR1 (%; 68.0 versus 50.0). We concluded that programmable fast-freezing method (FR3) improves the post-thaw in vitro quality and in vivo fertility of water buffalo spermatozoa.
Animal Reproduction Laboratory, Animal Sciences Institute, National Agricultural Research Center, Islamabad, Pakistan.
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Khalil Ur Rehman
Animal Reproduction Laboratory, Animal Sciences Institute, National Agricultural Research Center, Islamabad, Pakistan.
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SMH
Andrabi
Animal Reproduction Laboratory, Animal Sciences Institute, National Agricultural Research Center, Islamabad, Pakistan.
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Ahmed
Animal Reproduction Laboratory, Animal Sciences Institute, National Agricultural Research Center, Islamabad, Pakistan.
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SAH
Shah
D000177:Acrosome / Q000502:physiology*; D000818:Animals; D002020:Buffaloes; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology; D004272:DNA, Mitochondrial; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary; D005615:Freezing; D008297:Male; D051336:Mitochondrial Membranes / Q000502:physiology*; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility / Q000502:physiology; D013094:Spermatozoa / Q000502:physiology*
D003451:Cryoprotective Agents; D004272:DNA, Mitochondrial
buffalo spermatozoa; freezing rates; in vitro quality; in vivo fertility