Evaluation of antifreeze protein III for cryopreservation of Nili-Ravi (Bubalus bubalis) buffalo bull sperm

• Published in: Animal reproduction science 2014-07, Vol.148 (1-2), p.26-31
• Main Authors: Qadeer, S., Khan, M.A., Ansari, M.S., Rakha, B.A., Ejaz, R., Husna, A.U., Ashiq, M., Iqbal, R., Ullah, N., Akhter, S.
• Format: Article
• Language: English
• Subjects: Animals; Antifreeze Proteins, Type III - pharmacology; Bubalus bubalis; Buffalo bull sperm, Antifreeze proteins, Ice crystal; Buffaloes - physiology; Cryopreservation - veterinary; Cryoprotective Agents - pharmacology; Dose-Response Relationship, Drug; Fertility; Male; Semen Analysis; Semen Preservation - methods; Semen Preservation - veterinary; Sperm Motility - drug effects; Spermatozoa - cytology; Spermatozoa - drug effects; Spermatozoa - physiology
• ISSN: 0378-4320; 1873-2232
• DOI: 10.1016/j.anireprosci.2014.04.013

Lower fertility in buffaloes with frozen-thawed semen is attributed to sperm damage that is believed to be due to formation of ice crystals during freeze/thaw process. It was hypothesized that antifreeze proteins in the extender may improve the post thaw quality of buffalo bull sperm. For this purpose, two separate experiments were conducted to evaluate antifreeze proteins III (AFP III) at 0 (control), 0.1, 1 and 10μg/mL (Experiment I) and 0 (control), 0.01, 0.1 and 1μg/mL (Experiment II) for its effect on post thaw quality of buffalo bull semen. Semen was collected from three Nili-Ravi buffalo (Bubalus bubalis) bulls with artificial vagina (42°C) for three weeks (replicate) per experiment. For each experiment, qualifying ejaculates (6ejaculates/bull) were divided into four aliquots and diluted (at 37°C having 50×106sperm/mL) in tris-citric acid extender containing above mentioned concentrations of AFP III. Diluted semen was cooled to 4°C in 2h, equilibrated for 4h, filled in 0.5mL straws, kept over liquid nitrogen vapors for 10min and plunged in the liquid nitrogen. After 24h of storage, semen straws were thawed at 37°C for 30s to assess sperm progressive motility (SM), plasma membrane integrity (PMI), viability (live sperm with intact acrosome) and normal epical ridge (NAR). In experiment I, improvement (P0.05) in extenders containing different concentrations of AFP III and control in both of experiments. In conclusion addition of AFP III in the extender at 0.1μg/mL improved the progressive motility and plasma membrane integrity of cryopreserved buffalo bull semen.