Comparison of slow freezing and vitrification methods for Venda cockerel’s spermatozoa

Masindi L. Mphaphathi, Dibungi Luseba, Ben Sutherland, Tshimangadzo L. Nedambale

Agricultural Research Council, Animal Production Institute, Germplasm Conservation & Reproductive Biotechnologies, Irene, RSA.
Tshwane University of Technology, Faculty of Science, Department of Animal Sciences, Pretoria, RSA.
University of the Free State, Department of Animal, Wildlife and Grassland Sciences, Bloemfontein, RSA.

DOI: 10.4236/ojas.2012.23028                

Abstract

An improvement in avian semen cryopreservation is essential and has the potential to improve the cryo-gene banking efficiency. This study compared two cryopreservation methods (slow freezing and vitrification) and the effect of different thawing/warming temperatures (5℃, 25℃ and 41℃) on Venda cockerel’s spermatozoa. Semen samples from Venda cockerels were diluted with modified Kobidil+ extender supplemented with 8% dimethyl sulfoxide. Semen from each ejaculate was stained with nigrosin/eosin for viability examination. The cryopreserved samples were either slow cooled in 0.25 mL straw or vitrified in a solid surface vitrification (SSV) device. Semen straw or cryovial was stored in liquid nitrogen container. The straw or cryovial with sperm was thawed or warmed at 5?C, 25?C and 41℃ and analysed by a Computer-Aided Sperm Analysis (CASA). There was a significant difference in live/normal sperm between the semen donors. Cockerels spermatozoa cryopreserved by slow freezing (43%) and thawed at 5?C had a significantly higher survival and motility rate compared to vitrification (2.5%) method. In conclusion, there was higher rate of live/normal morphology sperm. Cryopreservation process reduces sperm motility and velocity rate regardless of cryoprevervation method and thawing or warming temperatures. However, slow freezing was a better method to maintain motility of spermatozoa following cryopreservation.

Keywords

Venda Cockerels; Dimethyl Sulfoxide; Sperm Motility; Cryopreservation

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Fulton, J.E. (2006) Avian genetic preservation: An industry perspective. Poultry Science, 85, 227-231.
[2]	Donoghue, A.M. and Wishart, G.J. (2000) Storage of poultry semen. Animal Reproduction Science, 62, 213- 232. doi:10.1016/S0378-4320(00)00160-3
[3]	Van Marle-Koeste, E., Hefer, C.A., Nel, L.H. and Groenen, M.A.M. (2008) Genetic diversity and population structure of locally adapted South African chicken lines: implications for conservation. South African Journal of Animal Science, 38, 271-281.
[4]	Long, J.A. (2008) Reproductive biotechnology and gene mapping: tools for conserving rare breeds of livestock. Reproduction of Domestic Animals, 43, 83-88. doi:10.1111/j.1439-0531.2008.01146.x
[5]	Blanco, J.M., Gee, G., Wildt, D.E. and Donoghue, A.M. (2000) Species variation in osmotic, cryoprotectant, and cooling rate tolerance in poultry, eagle and Peregrine falcon spermatozoa. Biology of Reproduction, 63, 1164- 1171. doi:10.1095/biolreprod63.4.1164
[6]	Purdy, P.H., Song, Y., Silversides, F.G. and Blackburn, H.D. (2009) Evaluation of glycerol removal techniques, cryoprotectants, and insemination methods for cryopreservation rooster sperm with implications of regeneration of breed or line or both. Poultry Science, 88, 2184-2191. doi:10.3382/ps.2008-00402
[7]	Williamson, R.G., Etchers, R.J., Reinhart, B.S. and Macpherson, J.W. (1981) The effect of cooling rate before freezing and the temperature of the semen upon addition of DMSO on the fertilizing capacity of chicken semen stored at –196?C. Reproduction of Nutrition and Development, 21, 1033-1042. doi:10.1051/rnd:19810801
[8]	Dinnyes, A., Liu, J. and Nedambale, T.L. (2007) Novel gamete storage. Reproduction of Fertility and Development, 19, 719-731. doi:10.1071/RD07035
[9]	Ozkavukcu, S. and Erdemli, E. (2000) Cryopreservation: basic knowledge and biophysical effects. Journal of Ankara Medical School, 4, 187-196.
[10]	Nedambale, T.L., Dinnyes, A., Yang, X. and Tian, C. (2004) Bovine blastocyst development in vitro: Timing, sex, and viability following vitrification. Biology of Reproduction, 71, 1671-1676. doi:10.1095/biolreprod.104.027987
[11]	Parksw, E.J., Heck, W.R. and Hardaswick, V. (1986) Cryopreservation of Peregrine falcon semen post-thaw dialysis to remove glycerol. Raptor Research, 20, 15-20.
[12]	Long, J.A. and Kulkarin, G. (2004) An effective method for improving the fertility of glycer-ol-exposed poultry semen. Poultry Science, 83, 1594-1601.
[13]	Lake, P.E. and Ravie, O. (1984) An exploration of cryoprotective compounds for fowl spermatozoa. British Poultry Science, 25, 145-150. doi:10.1080/13632758408454852
[14]	Sontakke, S.D., Umapathy, G., Sivaram, V., Kholkutel, S.D. and Shivaji, S. (2004) Semen characteristics, cryopreservation, and successful artificial insemination in the Blue rock pigeon (Columba livia). Theriogenology, 62, 139-153. doi:10.1016/j.theriogenology.2003.08.018
[15]	Burrows, H.W. and Quinn, J.P (1937) The collection of spermatozoa from the domestic fowl and turkey. Poultry Science, 16, 185-191. doi:10.3382/ps.0160019
[16]	EI-Gendy, E.A., Gad, A.Y. and Mostageer, A (2007) Sperm-mediated gene transfer in poultry. The relationship with cock sperm viability. Arab Journal of Biotechnology, 10, 1-12.
[17]	Lukaszewick, E., Jerysz, A., Partyka, A. and Siudzinska, A. (2008) Efficacy of evaluation of rooster sperm morphology using different staining methods. Research in Veterinary Science, 85, 583-588. doi:10.1016/j.rvsc.2008.03.010
[18]	Tselutin, K., Seigneurin, F. and Blesbois, E. (1999) Comparison of cryoprotectants and methods of cryopreservation of fowl spermatozoa. Poultry Science, 78, 586- 590.
[19]	Ansah, G.A., Segura, J.C. and Buckland, R.B (1985) Semen production, sperm quality, and their heritabilities as influenced by selection for fertility of frozen-thawed semen in the chicken. Poultry Science, 64, 1801-1803. doi:10.3382/ps.0641801
[20]	Gendy, E.A., Gad, A.Y. and Mostageer, A (2007) Sperm-mediated gene transfer in poultry. The relationship with cock sperm viability. Arab Journal of Biotechnology, 10, 1-12.
[21]	Rwuaan, J.S., Rekwot, P.I., Abdu, P.A., Eduvie, L.O. and Obidi, J.A (2009) Semen characteristics of vaccinated Shikabrown cocks challenged with a velogenic Newcastle disease virus. International Journal of Poultry Science, 8, 896-90. doi:10.3923/ijps.2009.896.901
[22]	Tuncer, P.B., Kinet, H. and Ozdogan, N (2008) Evaluation of some spermatological characteristics in Gerze cocks. Ankara University of Veterinarian Dergree, 55, 99- 102.
[23]	Blesbois, E., Grasseau, I., Seigneurin, F., Mignon-Grasteau, S., Saint-Jalme, M. and Milalon-Richard, M.M. (2008) Predictors of success of semen cryopreservation in chickens. Theriogenology, 69, 349-359. doi:10.1016/j.theriogenology.2007.09.019
[24]	Long, J.A., Bongalhardo, D.C., Pelaez, J., Saxena, S., Settar, P., O’Sullivan, N.P. and Fulton, J.E. (2010) Rooster semen cryopreservation:effect of pedigree line and male age on postthaw sperm function. Poultry Science, 89, 966-973. doi:10.3382/ps.2009-00227
[25]	Han, X.F., Niu, Z.Y., Liu, F.Z. and Yang, C.S. (2005) Effects of diluents, cryoprotectants, equilibration time and thawing temperature on cryopreservation of duck semen. Poultry Science, 4, 197-201. doi:10.3923/ijps.2005.197.201
[26]	Peirouvi, T., Farjah, G., Rad, J.S. and Novin, M.G. (2007) Vitrification induced apopto-sis in spermatozoa from fertile and subfertile men. Iran Journal of Reproductive Medicine, 5, 117-120.
[27]	Tselutin, K., Seigneurin, F. and Blesbois, E. (1999) Comparison of cryoprotectants and methods of cryopreservation of fowl sper-matozoa. Poultry Science, 78, 586- 590.
[28]	Siudzinska, A. and Lukaszewicz, E. (2008) The effect of breed on freezability of semen of fancy fowl. Animal Science Papers and Reports, 26, 331-340.
[29]	Orief, Y., Schultze-Mosgau, A., Dafopoulos, K., AI-Hasani, S. (2005) Vitrification: Will it replace the conventional gamete cryopreservation techniques? Middle East Fertility Society Journal, 10, 171-184.
[30]	Makhafola, M.B, Lehloenya., K.C., Mphaphathi, M.L. and Dinnyes, A. Nedambale, T.L. (2009) The effect of breed on the survivability and motility rate of cryopreserved cock semen. South African Journal of Animal Science, 39, 242-245.
[31]	Jalme, M.S., Lecoq, R., Seigneurin, F., Blesbois, E. and Plouzeau, E. (2003) Cryopreservation of semen from endangered pheasants:the first step towards a cryobank for endangered avian species. Therionelogy, 59, 875-888. doi:10.1016/S0093-691X(02)01153-6
[32]	Madedu, M., Berlinguer, F., Ledda, M., Leoni, G.G., Satta, V., Succu, S., Rotta, A., Pasciu, V., Zinellu, A., Muzzeddu, M., Carru, C. and Naitana, S. (2009) Ejaculate collection efficiency and post-thaw semen quality in wild-caught Griffon vultures from the Sardinian population. Reproductive Biology Endocrinology, 7, 1-11. doi:10.1186/1477-7827-7-18
[33]	Martin, G., Cagnon, N., Sabido, O., Sion, B., Grizard, G., Durand, P. and Levy, R. (2007) Kinetics of occurrence of some features of apoptosis during the cryopreservation process of bovine spermatozoa. Human Reproduction, 22, 380-388. doi:10.1093/humrep/del399
[34]	Ortega-Ferrulosa, C., Sotillio-Galan, Y., Varela-Fernandez, E., Gallardo-Bolanos, J.M., Muriel, A., Gonzalez-Fernandez, L., Tapia, J.A and Pena, F.J. (2008) Detection of “apoptosis-like” changes during the cryopreservation process in equine sperm. Journal of Andrology, 29, 213- 221. doi:10.2164/jandrol.107.003640
[35]	Waterhouse, K.E., Hofmo, P.O., Tverdal, A. and Miller, R.R. (2006) Within and between breed differences in freezing tolerance and plasma membrane fatty acid composition of boar sperm. The Journal of the Animal Society for Reproduction and Fertility, 131, 887-894. doi:10.1530/rep.1.01049
[36]	Defoin, L., Granados, A. and Donnay, I. (2007) Analysing motility parameters on fresh bull semen could help to predict resistance to freezing: A preliminary study. Reproduction in Domestic Animals, 43, 606-611. doi:10.1111/j.1439-0531.2007.00964.x