Cryopreservation of Swamp Buffalo Semen in Skim Milk Yolk-based Diluent with Two Different Cryoprotectants

The successful of artificial insemination (AI) in buffalo is still low. The quality of frozen semen determines the successful of AI program. Glycerol is widely used for cryopreservation of buffalo semen with unsatisfactory results due to its toxicity and antifertility properties. This study aimed to investigate the effect of glycerol and dimethylformamide (DMF) concentrations in skim milk egg yolk (SMEY)–based diluent on the quality of frozen-thawed swamp buffalo bull semen. Fresh semen was divided into eight aliquots. Each aliquot was diluted in SMEY containing 4%, 5%, 6%, and 7% glycerol and 4%, 5%, 6%, and 7% DMF. Diluted semen was packed into mini straw (0.25 mL), equilibrated at 4 °C for 4 hours, frozen in liquid nitrogen vapor for 10 minutes, and stored at liquid nitrogen container. The straws were thawed after 24 hours at 37 °C for 30 seconds for evaluation. The results showed that the post thawed sperm motility and recovery rate in SMEY extender containing 7% glycerol and 5% DMF were higher than the other glycerol or DMF concentrations. Post thawed sperm viability and plasma membrane integrity in SMEY containing 4%-6% DMF did not differ significantly, however both values were higher than those in 7% DMF. Post thawed sperm viability and plasma membrane integrity did not differ between SMEY containing 6% and 7% glycerol. However, both values were higher than those in 4% and 5% glycerol. It is concluded that 7% glycerol or 5% DMF is considered to be the most suitable cryoprotective agent for swamp buffalo semen cryopreservation.The successful of artificial insemination (AI) in buffalo is still low. The quality of frozen semen determines the successful of AI program. Glycerol is widely used for cryopreservation of buffalo semen with unsatisfactory results due to its toxicity and antifertility properties. This study aimed to investigate the effect of glycerol and dimethylformamide (DMF) concentrations in skim milk egg yolk (SMEY)–based diluent on the quality of frozen-thawed swamp buffalo bull semen. Fresh semen was divided into eight aliquots. Each aliquot was diluted in SMEY containing 4%, 5%, 6%, and 7% glycerol and 4%, 5%, 6%, and 7% DMF. Diluted semen was packed into mini straw (0.25 mL), equilibrated at 4 °C for 4 hours, frozen in liquid nitrogen vapor for 10 minutes, and stored at liquid nitrogen container. The straws were thawed after 24 hours at 37 °C for 30 seconds for evaluation. The results showed that the post thawed sperm motility and recovery rate in SMEY extender containing 7% glycerol and 5% DMF were higher than the other glycerol or DMF concentrations. Post thawed sperm viability and plasma membrane integrity in SMEY containing 4%-6% DMF did not differ significantly, however both values were higher than those in 7% DMF. Post thawed sperm viability and plasma membrane integrity did not differ between SMEY containing 6% and 7% glycerol. However, both values were higher than those in 4% and 5% glycerol. It is concluded that 7% glycerol or 5% DMF is considered to be the most suitable cryoprotective agent for swamp buffalo semen cryopreservation.

[1]  D. H. Kalhoro,et al.  Determination of Quality Characteristics of Kundhi Buffalo Bull Semen , 2016 .

[2]  I. Singh,et al.  New approaches in buffalo artificial insemination programs with special reference to India. , 2016, Theriogenology.

[3]  I. Blas,et al.  Use of soy milk combined with different cryoprotectants for the ram semen cryopreservation , 2016 .

[4]  Quan Liu,et al.  Chronic occupational N, N-dimethylformamide poisoning induced death: a case report , 2015, Forensic Science, Medicine, and Pathology.

[5]  W. Wolkers,et al.  Sperm Membrane Behaviour during Cooling and Cryopreservation. , 2015, Reproduction in domestic animals = Zuchthygiene.

[6]  Pradeep Kumar,et al.  Assessment of sperm damages during different stages of cryopreservation in water buffalo by fluorescent probes , 2016, Cytotechnology.

[7]  Md. Nazmul Hoque,et al.  Evaluation of ovsynch protocols for timed artificial insemination in water buffaloes in Bangladesh , 2014 .

[8]  D. Vecchio,et al.  Seasonal fluctuations in the response of Italian Mediterranean buffaloes to synchronization of ovulation and timed artificial insemination. , 2014, Theriogenology.

[9]  Shahverdi Abdolhossain,et al.  EFFECT OF EXTENDER AND EQUILIBRATION TIME ON POST THAW MOTILITY AND CHROMATIN STRUCTURE OF BUFFALO BULL (BUBALUS BUBALIS) SPERMATOZOA , 2014 .

[10]  R. I. Arifiantini,et al.  Effect of glycerol and dimethylformamide cryoprotectants on buck Etawah Crossbreed frozen semen using modified tris diluents. , 2013 .

[11]  A. Boediono,et al.  The Success Rate of Artificial Insemination Using Post-Thawed Spotted Buffaloes Epididymal Sperm , 2013 .

[12]  W. Maxwell,et al.  Dimethyl formamide improves the postthaw characteristics of sex-sorted and nonsorted stallion sperm. , 2013, Theriogenology.

[13]  K. Rohn,et al.  Osmotic Stress and Membrane Phase Changes During Freezing of Stallion Sperm: Mode of Action of Cryoprotective Agents1 , 2013, Biology of reproduction.

[14]  H. Rehman,et al.  Assessment of optimal osmotic pressure of citrate egg yolk extender for cryopreservation of buffalo bull (Bubalus bubalis) semen. , 2013 .

[15]  E. Celeghini,et al.  Effects of bovine sperm cryopreservation using different freezing techniques and cryoprotective agents on plasma, acrosomal and mitochondrial membranes , 2012, Andrologia.

[16]  Tae-Young Kang,et al.  Effect of Amides as a Cryoprotectant on Quality of Frozen-thawed Sperm in Korean Jeju Black Bull , 2012 .

[17]  F. Peña,et al.  Toxicity of glycerol for the stallion spermatozoa: effects on membrane integrity and cytoskeleton, lipid peroxidation and mitochondrial membrane potential. , 2012, Theriogenology.

[18]  Amrozi,et al.  Effect of glutathione and bovine seminal plasma in lactose extender on viability of swamp buffalo frozen semen. , 2012 .

[19]  G. L. Lima,et al.  Objective assessment of the cryoprotective effects of dimethylformamide for freezing goat semen. , 2011, Cryobiology.

[20]  A. R. Silva,et al.  Dimethylformamide as a cryoprotectant for canine semen diluted and frozen in ACP-106C. , 2011, Theriogenology.

[21]  F. Varago,et al.  Extenders containing dimethylformamide associated or not with glycerol are ineffective for ovine sperm cryopreservation. , 2011, Reproduction in domestic animals = Zuchthygiene.

[22]  Tae Hyun Kim,et al.  Clinical Outcomes of Occupational Exposure to N,N-Dimethylformamide: Perspectives from Experimental Toxicology , 2011, Safety and health at work.

[23]  R. I. Arifiantini,et al.  Motility and Viability of Friesian Holstein Spermatozoa in Three Different Extender Stored at 5oc , 2010 .

[24]  S. Akhter,et al.  Cryopreservation of buffalo (Bubalus bubalis) semen in Bioxcell extender. , 2010, Theriogenology.

[25]  R. Garnier,et al.  Dimethylformamide metabolism following self-harm using a veterinary euthanasia product , 2010, Clinical toxicology.

[26]  S. Atreja,et al.  Effects of adding taurine and trehalose to a tris-based egg yolk extender on buffalo (Bubalus bubalis) sperm quality following cryopreservation. , 2010, Animal reproduction science.

[27]  N. Carvalho,et al.  Fixed-time artificial insemination in buffalo , 2010 .

[28]  E. M. Madeira,et al.  Evaluation of amides and centrifugation temperature in boar semen cryopreservation. , 2008, Theriogenology.

[29]  N. Ahmed,et al.  Antagonist effect of DMSO on the cryoprotection ability of glycerol during cryopreservation of buffalo sperm. , 2007, Theriogenology.

[30]  H. Meryman,et al.  Cryopreservation of living cells: principles and practice , 2007, Transfusion.

[31]  D. Shieh,et al.  Mitochondrial DNA alterations in blood of the humans exposed to N,N-dimethylformamide. , 2007, Chemico-biological interactions.

[32]  I. Supriatna,et al.  Stallion semen cryopreservation using different cryoprotective agents on the skim milk trehalosa extender , 2007 .

[33]  D. Pegg.  Principles of cryopreservation. , 2007, Methods in molecular biology.

[34]  H. Rodríguez-Martínez,et al.  Seasonal variation in semen quality of swamp buffalo bulls (Bubalus bubalis) in Thailand. , 2007, Asian journal of andrology.

[35]  V. Paul,et al.  Efficacy of the Ovsynch protocol for synchronization of ovulation and fixed-time artificial insemination in Murrah buffaloes (Bubalus bubalis). , 2005, Theriogenology.

[36]  J. Blanco,et al.  Species Variation in Osmotic, Cryoprotectant, and Cooling Rate Tolerance in Poultry, Eagle, and Peregrine Falcon Spermatozoa1 , 2000, Biology of reproduction.

[37]  I. Gordon.  Controlled reproduction in cattle and buffaloes , 1999 .

[38]  H. Merker,et al.  Potency of monomethyl-, dimethylformamide and some of their metabolites to induce abnormal development in a limb Bud organ culture. , 1998, Toxicology in vitro : an international journal published in association with BIBRA.