Use of galactose to vitrify ram semen in straws
DOI:
https://doi.org/10.1590/1809-6891v22e-67525Abstract
Sperm vitrification is a technique with great potential for cryopreservation of genetic material, with superior effectiveness compared to conventional methods in some species. However, few studies have shown its efficiency with ejaculated sperm of rams and the use of galactose as an extracellular cryoprotectant during vitrification. This study aimed to evaluate the effect of galactose (0.01 M), with or without glycerol addition (3% and 7%) in a commercial extender (Steridyl® - control) for ram sperm cryopreservation in straws, comparing the classic freezing method and vitrification. Ejaculates from six breeding soundness Dorper rams were collected with an artificial vagina, aliquoted, individually diluted (100 × 106 sperm/mL) on extenders tested, loaded into 0.25 mL straws, and subjected to a classic freezing method or vitrification. Sperm kinematics, morphology, morphometry, viability, and physical and functional integrity of the sperm membrane were evaluated. The classic freezing method resulted in higher total and progressive motility than vitrification, as no motility was detected in vitrified samples after rewarming (p<0.05). The addition of galactose or glycerol to the commercial medium did not benefit both vitrification and the classic freezing method.
Keywords: Semen biotechnology; Ultra-rapid freezing; Cryoprotectants; Reproduction.
Downloads
References
Pegg DE. The history and principles of cryopreservation. Seminars in Reproductive Medicine. 2002;20(1):5–13. https://doi.org/10.1055/s-2002-23515.
Rumpf R. Avanços metodológicos na produção in vitro de embriões. Revista Brasileira de Zootecnia. 2007;36(suppl.0):229–233. https://doi.org/10.1590/S1516-35982007001000021.
Bittencourt RF, Oba E, Ribeiro Filho AL, Chalhoub M, Azevedo HC, Bicudo SD. Avanços na criopreservação do sêmen ovino I: diluidores e crioprotetores. Ciência Animal Brasileira. 2013;14(4):522-536. https://doi.org/10.5216/cab.v14I4.22964.
Carvalho AA, Faustino LR, Figueiredo JR, Rodrigues APR, Costa APR. Vitrificação: uma alternativa para a preservação de embriões e material genético de fêmeas mamíferas em criobancos. Acta Veterinaria Brasilica. 2011;5(3):236-248.
https://doi.org/10.21708/avb.2011.5.3.2321.
Matias MC, Melo MMS, Mendonça MSB, Sousa CWC, Costa Neto LF, Câmara DR. Vitrificação de espermatozoides em pequenos ruminantes. Revista Brasileira de Reprodução Animal. 2019;43(2):105-110.
http://cbra.org.br/portal/downloads/publicacoes/rbra/v43/n2/p105-110%20(RB776).pdf.
Vizuete G, Jiménez E, Agüera EI, Pérez-Marín CC. Impact of ultra-rapid freezing on the motility, morphology, viability and acrosome integrity of epididymal cat sperm diluted in sucrose-based extenders. Reproduction in Domestic Animals. 2014;49(1):e5–8. https://doi.org/10.1111/rda.12253.
Slabbert M, du Plessis SS, Huyser C. Large volume cryoprotectant-free vitrification: an alternative to conventional cryopreservation for human spermatozoa. Andrologia. 2015;47(5):594–599. https://doi.org/10.1111/and.12307.
Diaz-Jimenez M, Dorado J, Ortiz I, Consuegra C, Pereira B, Gonzalez-De Cara CA, Aguilera R, Mari G, Mislei B, Love CC, Hidalgo, M. Cryopreservation of donkey sperm using non-permeable cryoprotectants. Animal Reproduction Science. 2018; 189:103–109. https://doi.org/10.1016/j.anireprosci.2017.12.013.
Hidalgo M, Consuegra C, Dorado J, Diaz-Jimenez M, Ortiz I, Pereira B, Sanchez R, Crespo F. Concentrations of non-permeable cryoprotectants and equilibration temperatures are key factors for stallion sperm vitrification success. Animal Reproduction Science. 2018;196:91–98. https://doi.org/10.1016/j.anireprosci.2018.06.022.
Pipan MZ, Casal ML, Šterbenc N, Klun IV, Mrkun J. Vitrification using soy lecithin and sucrose: a new way to store the sperm for the preservation of canine reproductive function. Animals. 2020;10(4):653. https://doi.org/10.3390/ani10040653.
Pradiee J, Esteso MC, Castaño C, Toledano-Díaz A, Lopez-Sebastián A, Guerra R, Santiago-Moreno, J. Conventional slow freezing cryopreserves mouflon spermatozoa better than vitrification. Andrologia. 2017;49(3):e12629. https://doi.org/10.1111/and.12629.
Bóveda P, Esteso MC, Castaño C, Toledano-Díaz A, López-Sebastián A, Muñiz A, Prieto P, Mejía O, Ungerfeld R, Santiago-Moreno J. Slow and ultra-rapid freezing protocols for cryopreserving mouflon (Ovis musimon) and fallow deer (Dama dama) epididymal sperm. Animal Reproduction Science. 2018;192:193–199.
https://doi.org/10.1016/j.anireprosci.2018.03.010.
Martínez-Fresneda L, Castaño C, Bóveda P, Tesfaye D, Schellander K, Santiago-Moreno J, García-Vázquez FA. Epididymal and ejaculated sperm differ on their response to the cryopreservation and capacitation processes in mouflon (Ovis musimon). Scientific Reports. 2019;9:15659. https://doi.org/10.1038/s41598-019-52057-0.
Herrick JR, Wang C, Machaty Z. The effects of permeating cryoprotectants on intracellular free-calcium concentrations and developmental potential of in vitro-matured feline oocytes. Reproduction, Fertility and Development. 2016;28(5):599-607. https://doi.org/10.1071/rd14233.
Hinrichs K, Choi Y-H. Micromanipulation of equine blastocysts to allow vitrification. Reproduction, Fertility and Development. 2016;28(8):1092-1096. https://doi.org/10.1071/rd15389.
Chaytor JL, Tokarew JM, Wu LK, Leclère M, Tam RY, Capicciotti CJ, Guolla L, von Moos E, Findlay CS, Allan DS, Ben RN. Inhibiting ice recrystallization and optimization of cell viability after cryopreservation. Glycobiology. 2012; 22(1):123-33. https://doi.org/10.1093/glycob/cwr115.
Colégio Brasileiro de Reprodução AnimaL. Manual para exame andrológico e avaliação de sêmen animal. 3a. Belo Horizonte: CBRA; 2013. 104 p.
Aisen EG, Medina VH, Venturino A. Cryopreservation and post-thawed fertility of ram semen frozen in different trehalose concentrations. Theriogenology. 2002; 57(7):1801-1808. https://doi.org/10.1016/S0093-691X(02)00653-2.
Oliveira IRS, Alves HM, Castelo TS, Bezerra FSB, Bezerra ACDS, Silva AR. Correlações entre o teste hiposmótico e a avaliação clássica do sêmen de caprinos. Ciência Animal Brasileira. 2013;14(2):216–221. https://doi.org/10.5216/cab.v14i2.12921.
Masoud R, Sharafi M, Shahnenh AZ, Towhidi A, Kohram H, Zhandi M, Esmaeili V, Shahverdi A. Effect of dietary fish oil supplementation on ram semen freeze ability and fertility using soybean lecithin- and egg-yolk-based extenders. Theriogenology. 2016; 86(6):1583-1588. http://dx.doi.org/10.1016/j.theriogenology.2016.05.018.
Diaz-Jimenez M, Dorado J, Pereira B, Ortiz I, Consuegra C, Bottrel M, Ortiz E, Hidalgo M. Vitrification in straws conserves motility features better than spheres in donkey sperm. Reproduction in Domestic Animals. 2018;53(Suppl.2):56–58. https://doi.org/10.1111/rda.13256.
Alquézar-Baeta C, Gimeno-Martos S, Miguel-Jiménez S, Santolaria P, Yániz J, Palacín I, Casao A, Cebrián-Pérez JA, Muiño-Blanco T, Pérez-Pé R. OpenCASA: A new open-source and scalable tool for sperm quality analysis. PLOS Computational Biology. 2019;15(1):e1006691. https://doi.org/10.1371/journal.pcbi.1006691.
Arando A, Gonzalez A, Delgado JV, Arrebola FA, Perez-Marín CC. Storage temperature and sucrose concentrations affect ram sperm quality after vitrification. Animal Reproduction Science. 2017;181:175–185. https://doi.org/10.1016/j.anireprosci.2017.04.008.
Isachenko E, Isachenko V, Katkov II, Dessole S, Nawroth F. Vitrification of mammalian spermatozoa in the absence of cryoprotectants: from past practical difficulties to present success. Reproductive BioMedicine Online. 2003;6(2):191–200. https://doi.org/10.1016/s1472-6483(10)61710-5.
Jiménez-Rabadán P, García-Álvarez O, Vidal A, Maroto-Morales A, Iniesta-Cuerda M, Ramón M, del Olmo E, Fernández-Santos R, Garde JJ, Soler AJ. Effects of vitrification on ram spermatozoa using free-egg yolk extenders. Cryobiology. 2015;71(1):85–90. https://doi.org/10.1016/j.cryobiol.2015.05.004.
Fiser PS, Fairfull RW. The effect of glycerol concentration and cooling velocity on cryosurvival of ram spermatozoa frozen in straws. Cryobiology. 1984;21(5):542–551. https://doi.org/10.1016/0011-2240(84)90053-1.
Petrunkina AM, Waberski D, Günzel-Apel AR, Töpfer-Petersen E. Determinants of sperm quality and fertility in domestic species. Reproduction. 2007;134(1):3–17. https://doi.org/10.1530/rep-07-0046.
Arando A, Delgado JV, Arrebola FA, León JM, Alcalá CJ, Pérez-Marín CC. Vitrification induces critical subcellular damages in ram spermatozoa. Cryobiology. 2019;87:52–59. https://doi.org/10.1016/j.cryobiol.2019.02.005.
Curry MR, Watson PF. Osmotic effects on ram and human sperm membranes in relation to thawing injury. Cryobiology. 1994;31(1):39–46. https://doi.org/10.1006/cryo.1994.1005.
Published
How to Cite
Issue
Section
License
Copyright (c) 2021 CIÊNCIA ANIMAL BRASILEIRA (Brazilian Animal Science)
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g. in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
