1. bookVolume 20 (2020): Issue 3 (July 2020)
Journal Details
License
Format
Journal
eISSN
2300-8733
First Published
25 Nov 2011
Publication timeframe
4 times per year
Languages
English
access type Open Access

Improving the Rabbit Semen Cryopreservation Protocol: Comparison Between Two Extenders and Inseminating Doses

Published Online: 01 Aug 2020
Volume & Issue: Volume 20 (2020) - Issue 3 (July 2020)
Page range: 887 - 898
Received: 28 Oct 2019
Accepted: 14 Feb 2020
Journal Details
License
Format
Journal
eISSN
2300-8733
First Published
25 Nov 2011
Publication timeframe
4 times per year
Languages
English
Abstract

This study has been designed to optimize the semen freezing protocol in rabbits, in this regard we compared a Tris-citrate-glucose (TCG) extender with a commercial one (Cortalap®), that to the best of our knowledge has never been used up to now on the in vitro freezability and fertilizing ability of cryopreserved rabbit semen. Two different inseminating semen doses were considered. Five pooled semen samples were divided into two subsamples and each of them were diluted to a ratio 1:1 (v:v) with a freezing extender composed of TCG or Cortalap® containing 16% of dimethylsulfoxide and 0.1 mol/L of sucrose. The extended semen was filled into 0.25 mL plastic straws and frozen above a liquid nitrogen surface. After thawing (50°C/10 seconds) we determined sperm motility, viability, membrane functionality, acrosome and DNA integrity. Our results showed that the Cortalap® extender significantly improved the in vitro post-thaw sperm quality, in comparison to TCG. When we compared the extenders in vivo, no significant differences in the reproductive performances were observed independently by inseminating doses used. In this study we demonstrated that Cortalap® extender can be used as an alternative to TCG. Thus, the Cortalap® being a ready to use extender, implies a reduction of time, mistakes and microbial contaminations during its preparation. This discovery results as significant because it provides beyond an important contribution to the creation of the first Italian semen cryobank of rabbit breeds and also for livestock rabbit farms based on artificial insemination (AI) program.

Keywords

Arriola J., Foote R.H. (2001). Accessory sperm as an indication of fertilizing ability of rabbit spermatozoa frozen in egg yolk acetamide with detergent. J. Androl., 22: 458–463.Search in Google Scholar

Blesbois E. (2007). Current status in avian semen cryopreservation. Worlds Poult. Sci. J., 63: 213–222.Search in Google Scholar

Cortell C., Viudes de Castro M.P. (2008). Effect of gelatin addition to freezing extender on rabbit semen parameters and reproductive performance. In: Xiccato, G. (Ed.), 9th World Rabbit Congress, 327–332.Search in Google Scholar

Di Iorio M., Manchisi A., Rocco M., Chrenek P., Iaffaldano N. (2014). Comparison of different extenders on the preservability of rabbit semen stored at 5°C for 72 hours. Ital. J Anim. Sci, 13: 710–714.Search in Google Scholar

Di Iorio M., Colonna M.A., Miranda M., Principe P., Schiavitto M., Manchisi A., Iaffaldano N. (2018). Initial cooling time before freezing affects post-thaw quality and reproductive performance of rabbit semen. Anim. Sci. J., 89(9): 1240–1244.Search in Google Scholar

FAO. 2007. The State of the World’s Animal Genetic Resources for Food and Agriculture. Commission on Genetic Resources for Food and Agriculture. Food and Agriculture Organization of the United Nations, Rome, Italy.Search in Google Scholar

Foote R.H., Bratton R.W. (1950). The fertility of bovine semen in extenders containing sulfanilamide penicillin, streptomycin, and polymyxin. J. Dairy Sci. 33: 544–547.Search in Google Scholar

Gandini L., Lombardo F., Lenzi A., Spano M., Dondero F. (2006). Cryopreservation and sperm DNA integrity. Cell Tissue Bank., 7(2): 91–98.Search in Google Scholar

Hall S.E., Negus C., Johinke D., Bathgate R. (2017). Adjusting cryodiluent composition for improved post-thaw quality of rabbit spermatozoa. PLoS ONE, 12(4), e0175965.10.1371/journal.pone.0175965Search in Google Scholar

Iaffaldano N., Di Iorio M., Rosato M.P. (2012). The cryoprotectant used, its concentration, and the equilibration time are critical for the successful cryopreservation of rabbit sperm: Dimethylacetamide versus dimethylsulfoxide. Theriogenology, 78: 1381–1389.Search in Google Scholar

Iaffaldano N., Di Iorio M., Rosato M.P., Manchisi A. (2014). Cryopreservation of rabbit semen using non-permeable cryoprotectants: Effectiveness of different concentrations of low-density lipoproteins (LDL) from egg yolk versus egg yolk or sucrose. Anim. Reprod. Sci., 151: 220–228.Search in Google Scholar

Iaffaldano N., Di Iorio M., Cerolini S., Manchisi A. (2016). Overview of turkey semen storage: focus on cryopreservation – a review. Ann. Anim. Sci., 16(4): 961–97410.1515/aoas-2016-0026Search in Google Scholar

Jochová M., Novák K., Kott T., Zdeněk V., Majzlíka I., Tůmová E. (2017). Genetic characterization of Czech local rabbit breeds using microsatellite analysis. Livest. Sci., 201: 41–49.Search in Google Scholar

Kuliková B., Di Iorio M., Kubovicova E., Kuželová L., Iaffaldano N., Chrenek P. (2015). The cryoprotective effect of Ficoll on the rabbit spermatozoa quality. Zygote, 23: 785–794.Search in Google Scholar

Lavara R., Mocé E., Baselga M., Vicente, J.S. (2017). Freezability genetics in rabbit semen. Theriogenology, 102: 54–58.Search in Google Scholar

Layek S.S., Mohanty T.K., Kumaresan A., Parks J.E. (2016). Cryopreservation of bull semen: Evolution from egg yolk based to soybean based extenders. Anim. Reprod. Sci., 172: 1–9.Search in Google Scholar

Leroy G., Danchin-Burge C., Verrier E. (2011). Impact of the use of cryobank samples in a selected cattle breed: a simulation study. Genet. Sel. Evol., 43(1): 36.Search in Google Scholar

Lomeo A.M., Giambersio A.M. (1991) ‘Water-test’: a simple method to assess sperm-membrane integrity. Int. J. Androl., 14(4): 278–282.Search in Google Scholar

Maeda T., Liu E., Nishijima K., Tanaka M., Yamaguchi S., Morimoto M., Watanabe T., Fan J., Kitajima S. (2012). Effect of the primary cooling rate on the motility and fertility of frozen-thawed rabbit spermatozoa. World Rabbit Sci., 20: 65–70.Search in Google Scholar

Mendoza C., Correras A., Moos J., Tesarik J. (1992). Distinction between true acrosome reaction and degenerative acrosome loss by a one–step staining method using Pisum sativum agglutinin. J. Reprod. Infertil., 95: 755–763.Search in Google Scholar

Mocé E., Lavara R., Vicente J.S. (2003a). Effect of an asynchrony between ovulation and insemination on the results obtained after insemination with fresh or frozen sperm in rabbits. Anim. Reprod. Sci., 75: 107–118.10.1016/S0378-4320(02)00228-2Search in Google Scholar

Mocé E., Vicente J.S., Lavara R. (2003b). Effect of freezing–thawing protocols on the performance of semen from three rabbit lines after artificial insemination. Theriogenology, 60: 115–123.10.1016/S0093-691X(02)01329-8Search in Google Scholar

Mocé E., Vicente J.S. (2009). Rabbit sperm cryopreservation: A review. Anim. Reprod. Sci., 110: 1–24.Search in Google Scholar

Mocé E., Lavara R., Vicente J.S. (2010). Effect of cooling rate to 5°C, straw size and farm on fertilizing ability of cryopreserved rabbit sperm. Reprod. Domest. Anim., 45(5): e1–e7.Search in Google Scholar

Mocé E., Blanch E., Talaván A., Viudes de Castro M.P. (2014). Reducing the time rabbit sperm are held at 5°C negatively affects their fertilizing ability after cryopreservation. Theriogenology, 82: 1049–1053.Search in Google Scholar

Nishijima K., Kitajima S., Koshimoto C., Morimoto M., Watanabe T., Fan J., Matsuda Y. (2015). Motility and fertility of rabbit sperm cryopreserved using soybean lecithin as an alternative to egg yolk. Theriogenology, 84: 1172–1175.Search in Google Scholar

Rosato M.P., Iaffaldano N. (2011). Effect of chilling temperature on the long-term survival of rabbit spermatozoa held either in a tris based or a jellified extender. Reprod. Domest. Anim., 46(2): 301–308.Search in Google Scholar

Salamon S., Maxwell W.M.C. (2000). Storage of ram semen. Anim. Reprod. Sci. 62: 77–111.Search in Google Scholar

Viudes de Castro M.P., Mocé E., Vicente J.S., Marco-Jiménez F., Lavara R. (2005). In vitro evaluation of in vivo fertilizing ability of frozen rabbit semen. Reprod. Domest. Anim., 40: 136–140.Search in Google Scholar

Viudes de Castro M.P., Lavara R., Safaa H.M., Marco-Jiménez F., Mehaisen G.M.K., Vicente J.S. (2014). Effect of freezing extender composition and male line on semen traits and reproductive performance in rabbits. Animal, 8: 765–770.Search in Google Scholar

Zaniboni L., Cassinelli C., Mangiagalli M.G., Gliozzi T.M., Cerolini S. (2014). Pellet cryopreservation for chicken semen: effects of sperm working concentration, cryoprotectant concentration, and equilibration time during in vitro processing. Theriogenology, 82(2): 251–258.Search in Google Scholar

Zhu Z., Fan X., Lv Y., Zhang N., Fan C., Zhang P., Zeng W. (2015). Vitamin E analogue improves rabbit sperm quality during the process of cryopreservation through its antioxidative action. PLoS ONE, 10(12): e0145383.Search in Google Scholar

Zhu Z., Ren Z., Fan X., Pan Y., Lv S., Pan C., Lei A., Zeng W. (2017). Cysteine protects rabbit spermatozoa against reactive oxygen species-induced damages. PLoS ONE, 12(7): e0181110.Search in Google Scholar

Recommended articles from Trend MD

Plan your remote conference with Sciendo