Effect of Extender and Equilibration Time on Post Thaw Motility and Chromatin Structure of Buffalo Bull (Bubalus Bubalis) Spermatozoa
Objective: The aim of the present study was to investigate the effects of four equilibration times (2, 4, 8 and 16 hours) and two extenders (tris or Bioxcell®) on cryopreservation of buffalo semen. Materials and Methods: In this experimental study, split pooled ejaculates (n=4), possessing more...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Royan Institute (ACECR), Tehran
2014-04-01
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| Series: | Cell Journal |
| Subjects: | |
| Online Access: | http://celljournal.org/library/upload/article/af_52384335443224333224357735536427458442226-Abdolhossain%20Shahverdi-phone.pdf |
| Summary: | Objective: The aim of the present study was to investigate the effects of four equilibration
times (2, 4, 8 and 16 hours) and two extenders (tris or Bioxcell®) on cryopreservation
of buffalo semen.
Materials and Methods: In this experimental study, split pooled ejaculates (n=4), possessing
more than 70% visual sperm motility were divided in two aliquots and diluted in Bioxcell
® and tris-citric egg yolk (TCE) extenders. Semen was cooled to 4˚C within 2 hours,
equilibrated at 4˚C for 2, 4, 8 and 16 hours, then transferred into 0.5 ml French straws,
and frozen in a programmable cell freezer before being plunged into liquid nitrogen. Postthaw
motility characteristics, plasma membrane integrity, acrosome morphology and DNA
integrity of the buffalo sperm were studied after thawing.
Results: There were significant interactions between equilibration times and extenders
for sperm motility and membrane integrity. Post thaw sperm motility (PMOT), progressive
motile spermatozoa (PROG), plasma membrane integrity (PMI) and normal apical ridge
(NAR) measures were lower for sperm equilibrated for 2 hours in both TCE and Bioxcell®
extender compared to others equilibration times. PMOT, PMI and NAR for sperm equilibrated
for 4, 8 and 16 hours showed no significant differences in either extender, although
PROG measures were superior in Bioxcell® compared to TCE at all equilibration times
(p<0.05). Kinematic parameters such as average path velocity, curvilinear velocity and
linearity in the Bioxcell® extender were superior to those in the TCE extender studied. In
contrast to motility and viability, the DNA integrity of post thaw spermatozoa remained
unaffected by different equilibration times.
Conclusion: Equilibration time is necessary for preservation of the motility and integrity of
buffalo sperm membranes. Equilibration times of over than 2 hours resulted in the greatest
preservation of total semen parameters during cryopreservation. There were no significant
interactions between equilibration times |
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| ISSN: | 2228-5806 2228-5814 |