A Comparison of Freezing Methods and Diluents Types on Post-Thaw Sperm Quality of Ram Sperm
DOI:
https://doi.org/10.3329/ralf.v7i2.48863Keywords:
Diluents, Freezing methods, Cryopreservation, Ram semenAbstract
The effect of freezing methods and diluents types on post-thaw sperm quality of Bangladeshi ram semen was studied. Two freezing methods and three diluents was tested as pooling effects (freezing methods or diluents) on post-thaw sperm parameters; sperm motility (SM), viability (SV), plasma membrane integrity (SPMI) and acrosome integrity (SAI), respectively. From selected ten rams, eight ejaculates were used for each freezing group (freezing methods × diluents). Semen samples were diluted by using two-steps for hand-made tris-based diluents (20% egg yolk): D1 (7% glycerol) and D2 (5% glycerol), and one-step dilution for commercial diluents: D3 (Triladyl®) at 35°C. After 4h of equilibration of temperature at 5°C, diluted semen samples was aspirated into 0.25 mL straws, and sealed. Straws were frozen in liquid nitrogen (LN2) vapour using two methods: F1 (manually in Styrofoam box, using three-steps method; +5°C to -80°C at -11.33°C/min, -80°C to -120°C at -26.66°C/min, and -120°C to -140°C at - 13.33°C/min) and F2 (programmable bio-freezer, using two-steps method; +5°C to -100°C at - 20°C/min and -100°C to -140°C at -10°C/min). Two semen straws from each batch were evaluated (37°C for 20 sec) for sperm parameters. In pool effects between freezing methods; SAI differed significantly (P < 0.001). The SM (56%) and SV (72%) were observed competitive. However, SPMI (67.58 ± 2.02%) and SAI (76.13 ± 1.42%) were higher in F1. Among diluents, SM (P < 0.006), SV (P < 0.008), SPMI (P < 0.012) and SAI (P < 0.019) differed significantly. The SM (61.25 ± 1.80%), SV (77.13 ± 1.47%), SPMI (68.31 ± 1.91%) and SAI (74.75 ± 1.64%) were highest in D3. In conclusion, the combination of manual freezing (three-steps) and handmade tris-based diluents (20% egg yolk, 5% glycerol) is suitable and sustainable method for cryopreservation of ram semen.
Res. Agric., Livest. Fish.7(2): 235-241, August 2020
Downloads
19
28
Downloads
Published
How to Cite
Issue
Section
License
Creative Commons
All RALF articles are published under the terms of the Creative Commons Attribution License 4.0 International License. Readers can copy, distribute, transmit and adapt the work provided the original work and source is appropriately cited.
Copyright
Submission of a manuscript implies that authors have met the requirements of the editorial policy and publication ethics. Authors retain the copyright of their articles published in the journal. However, authors agree that their articles remain permanently open access under the terms of the Creative Commons Attribution License 4.0 International License.