Freezability of Andalusian donkey (Equus asinus) spermatozoa: effect of extenders and permeating cryoprotectants
D. Acha A , M. Hidalgo A , I. Ortiz A , M. J. Gálvez A , J. J. Carrasco B , V. Gómez-Arrones B and J. Dorado A C
+ Author Affiliations
- Author Affiliations
A Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Cordoba, Campus de Rabanales (Edif. Hospital Clínico Veterinario), Ctra. Madrid-Cádiz, km 396, 14071 Córdoba, Spain.
B Equine Reproduction Center, Centro de Selección y Reproducción Animal, (CENSYRA-Extremadura Government), Camino Santa Engracia, S/N (Estación Pecuaria), 06007 Badajoz, Spain.
C Corresponding author. Email: jdorado@uco.es
Reproduction, Fertility and Development 28(12) 1990-1998 https://doi.org/10.1071/RD14449
Abstract
The aim of this study was to compare the effect of two semen extenders and four permeating cryoprotectants on post-thaw sperm quality of Andalusian donkeys. First, 32 ejaculates were pooled, split and frozen in either Gent B or INRA 96 with egg yolk and glycerol. Second, 12 pooled semen samples were simultaneously frozen in Gent B (glycerol) or Gent A containing ethylene glycol (EG; 1 or 1.5%) or dimethyl sulfoxide (DMSO; 1.5 or 2%). Finally, nine pooled samples were simultaneously cryopreserved in Gent A containing 1% EG (as control), dimethylformamide (DMFA; 1 or 2.5%) or a combination of 1% EG and 1.5% DMFA. Gent B yielded a higher (P < 0.01) post-thaw sperm motility than modified INRA96. EG 1% increased the sperm membrane integrity (P < 0.001), whereas DMSO affected sperm motility and membrane integrity (P < 0.001). DMFA 2.5% yielded higher (P < 0.001) values for sperm motility and membrane integrity. We concluded that Gent B improves in vitro post-thaw sperm quality of donkey spermatozoa, but the replacement of glycerol with 1% EG or 2.5% DMFA increased sperm protection against cryodamage. The use of DMSO for freezing donkey semen was unsuccessful and a toxic effect is suspected. These extenders should be included in the pre-freeze test for each donkey.
Additional keywords: freezing extender, frozen-thawed semen, sperm quality.
References
Alvarenga, M. A., Keith, S. L., Graham, J. K., Landim-Alvarenga, F. C., and Squires, E. L. (2000a). Alternative cryoprotectors for freezing stallion semen. In ‘Proceedings of the 14th International Congress on Animal Reproduction, Stockholm, 2–6 July 2000’. (Ed(s) M. Forsberg, T. Greve, H. Gustafsson, T. Katila, H. Kindahli, E. Ropstad.) p. 73. (International Congress on Animal Reproduction – ICAR: Stockholm, Sweden.)Alvarenga, M. A., Landim-Alvarenga, F. C., Moreira, R. M., and Cesarino, M. M. (2000b). Acrosomal ultrastructure of stallion spermatozoa cryopreserved with ethylene glycol using two packaging systems. Equine Vet. J. 32, 541–545.
| Acrosomal ultrastructure of stallion spermatozoa cryopreserved with ethylene glycol using two packaging systems.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3M%2Fntl2isA%3D%3D&md5=7b834c0b34011addcc69a2052d4e286bCAS | 11093629PubMed |
Alvarenga, M. A., Papa, F. O., Landim-Alvarenga, F. C., and Medeiros, A. S. L. (2005). Amides as cryoprotectants for freezing stallion semen: a review. Anim. Reprod. Sci. 89, 105–113.
| Amides as cryoprotectants for freezing stallion semen: a review.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXhtVart77L&md5=71610e1670f828cfe86f3820971ab4c0CAS | 16099609PubMed |
Alvarez, J. G., and Storey, B. T. (1993). Evidence that membrane stress contributes more than lipid peroxidation to sublethal cryodamage in cryopreserved human spermatozoa: glycerol and other polyols as sole cryoprotectant. J. Androl. 14, 199–209.
| 1:CAS:528:DyaK2cXhtVegsbc%3D&md5=53d24436fea09bd94da19a1eb9abbb4fCAS | 8407576PubMed |
Alvarez, A. L., Serres, C., Crespo, F., Mateos, E., and Gómez-Cuétaral, C. (2004). Cryopreservation of Zamorano–Leonés donkey spermatozoa using different amides. Reprod. Domest. Anim. 39, 267.
Álvarez, A. L., Gómez-Cuétara, C., and Graham, J. K. (2005). Effect of cholesterol-loaded cyclodextrin and cryoprotectants on the cryosurvival of stallion spermatozoa. Reprod. Domest. Anim. 40, 375.
Álvarez, A. L., Serres, C., Torres, P., Crespo, F., Mateos, E., and Gómez-Cuétara, C. (2006). Effect of cholesterol-loaded cyclodextrin on the cryopreservation of donkey spermatozoa. Anim. Reprod. Sci. 94, 89–91.
Andrabi, S. M. H., and Maxwell, W. M. C. (2007). A review on reproductive biotechnologies for conservation of endangered mammalian species. Anim. Reprod. Sci. 99, 223–243.
| A review on reproductive biotechnologies for conservation of endangered mammalian species.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXjslGktbc%3D&md5=29f45fd87b3c77b0781730795b97b717CAS |
Ball, B. A., and Vo, A. (2001). Osmotic tolerance of equine spermatozoa and the effects of soluble cryoprotectants on equine sperm motility, viability and mitochondrial membrane potential. J. Androl. 22, 1061–1069.
| 1:CAS:528:DC%2BD3MXotlyns7o%3D&md5=ede1e87a0d52d407ab9e58c267ea7409CAS | 11700853PubMed |
Brito, L. F. (2007). Evaluation of stallion sperm morphology. Clinical Techniques in Equine Practice 6, 249–264.
| Evaluation of stallion sperm morphology.Crossref | GoogleScholarGoogle Scholar |
Bucci, D., Spinaci, M., Vallorani, C., Contri, A., Carluccio, A., Isani, G., Tamanini, C., and Galeati, G. (2010). Detection and localisation of GLUTs 1, 2, 3 and 5 in donkey spermatozoa. Reprod. Domest. Anim. 45, e217–e220.
| 1:CAS:528:DC%2BC3cXhtlSgtbzN&md5=df88267bb5dd0d1b3764daf41bfb4764CAS | 19930135PubMed |
Canisso, I. F., Souza, F. A., and Ortigoza Escobar, J. M. (2008). Congelamiento de semen de burro (Equus asinus). Rev. investig. vet. Perú 19, 113–125.
Canisso, I. F., Carvalho, G. R., Morel, M. D., Ker, P. G., Rodrigues, A. L., Silva, E. C., and Da Silva, M. A. C. (2011). Seminal parameters and field fertility of cryopreserved donkey jack semen after insemination of horse mares. Equine Vet. J. 43, 179–183.
| Seminal parameters and field fertility of cryopreserved donkey jack semen after insemination of horse mares.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3Mrjt12jug%3D%3D&md5=8f9849e154d1157ec7497ef1b51f2677CAS | 21592212PubMed |
Chenier, T., Merkies, K., Leibo, S., Plante, C., and Johnson, W. (1998). Evaluation of cryoprotective agents for use in the cryopreservation of equine spermatozoa. In ‘Proceedings of the 44th Annual Convention of the American Association of Equine Practitioners, Baltimore, 6–9 December 1998’. (Ed(s) Lexington, KY: American Association of Equine Practitioners – AAEP.) pp. 5–6. (American Association of Equine Practitioners – AAEP: Baltimore, MD.)
Contri, A., Gloria, A., Robbe, D., De Amicis, I., and Carluccio, A. (2012a). Characteristics of donkey spermatozoa along the length of the epididymis. Theriogenology 77, 166–173.
| Characteristics of donkey spermatozoa along the length of the epididymis.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC38%2FlvFaqsA%3D%3D&md5=23b0d5183e32bc0162988436abc19723CAS | 21872312PubMed |
Contri, A., Gloria, A., Robbe, D., Sfirro, M. P., and Carluccio, A. (2012b). Effect of sperm concentration on characteristics of frozen–thawed semen in donkeys. Anim. Reprod. Sci. 136, 74–80.
| Effect of sperm concentration on characteristics of frozen–thawed semen in donkeys.Crossref | GoogleScholarGoogle Scholar | 23182476PubMed |
Cortés-Gutiérrez, E. I., Crespo, F., Gosálvez, A., Dávila-Rodríguez, M. I., López-Fernández, C., and Gósalvez, J. (2008). DNA fragmentation in frozen spermatozoa of Equus asinus: Zamorano–Leonés, a breed at risk of extinction. Theriogenology 69, 1022–1032.
| DNA fragmentation in frozen spermatozoa of Equus asinus: Zamorano–Leonés, a breed at risk of extinction.Crossref | GoogleScholarGoogle Scholar | 18367243PubMed |
DAD-IS-FAO (2013) ‘Information System Domestic Animal Diversity’. (Food and Agriculture Organization of the United Nations – FAO.)
Dorado, J., Acha, D., Ortiz, I., Gálvez, M. J., Carrasco, J. J., Díaz, B., Gómez-Arrones, V., Calero-Carretero, R., and Hidalgo, M. (2013). Relationship between conventional semen characteristics, sperm motility patterns and fertility of Andalusian donkeys (Equus asinus). Anim. Reprod. Sci. 143, 64–71.
| Relationship between conventional semen characteristics, sperm motility patterns and fertility of Andalusian donkeys (Equus asinus).Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC2c7ks1alsA%3D%3D&md5=a812ac9d66a9d7a6b09d983ea2c9f496CAS | 24210876PubMed |
Dorado, J., Acha, D., Ortiz, I., Gálvez, M., Carrasco, J., Gómez-Arrones, V., Calero-Carretero, R., and Hidalgo, M. (2014). Effect of extender and amino-acid supplementation on sperm quality of cooled–preserved Andalusian donkey (Equus asinus) spermatozoa. Anim. Reprod. Sci. 146, 79–88.
| Effect of extender and amino-acid supplementation on sperm quality of cooled–preserved Andalusian donkey (Equus asinus) spermatozoa.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXjsF2gsbk%3D&md5=6f84c602a508fb744889001f830c1e40CAS | 24602506PubMed |
Flores, E., Taberner, E., Rivera, M., Peña, A., Rigau, T., Miró, J., and Rodríguez-Gil, J. (2008). Effects of freezing–thawing on motile sperm subpopulations of boar and donkey ejaculates. Theriogenology 70, 936–945.
| Effects of freezing–thawing on motile sperm subpopulations of boar and donkey ejaculates.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD1crnsVOrsw%3D%3D&md5=e691bc25c2a280efcac66405fad528a6CAS | 18602684PubMed |
Glazar, A. I., Mullen, S. F., Liu, J., Benson, J. D., Critser, J. K., Squires, E. L., and Graham, J. K. (2009). Osmotic tolerance limits and membrane permeability characteristics of stallion spermatozoa treated with cholesterol. Cryobiology 59, 201–206.
| Osmotic tolerance limits and membrane permeability characteristics of stallion spermatozoa treated with cholesterol.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhtVKgu7nE&md5=4a7298a3c67ea00c18dd05774bb3459bCAS | 19646432PubMed |
Gomes, G. M., Jacob, J. C. F., Medeiros, A. S. I., Papa, F. O., and Alvarenga, M. A. (2002). Improvement of stallion spermatozoa preservation with alternative cryoprotectants for the Mangalarga Marchador breed. Theriogenology 58, 277–279.
| 1:CAS:528:DC%2BD38XkvFynurs%3D&md5=9f881356133f0e146785cecf986fecb6CAS |
Henry, M., Snoeck, P. P. N., and Cottorello, A. C. P. (2002). Post-thaw spermatozoa plasma membrane integrity and motility of stallion semen frozen with different cryoprotectants. Theriogenology 58, 245–248.
| 1:CAS:528:DC%2BD38XkvFyntbc%3D&md5=4a5d73a887d18e53608e015f59edbc4fCAS |
Hoffmann, N., Morandini, C., and Sieme, H. (2010). Determination of the optimal concentration of cryoprotectant agent for freezing stallion spermatozoa. Anim. Reprod. Sci. 121, S160–S162.
Hoffmann, N., Oldenhof, H., Morandini, C., Rohn, K., and Sieme, H. (2011). Optimal concentrations of cryoprotective agents for semen from stallions that are classified ‘good’ or ‘poor’ for freezing. Anim. Reprod. Sci. 125, 112–118.
| Optimal concentrations of cryoprotective agents for semen from stallions that are classified ‘good’ or ‘poor’ for freezing.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXnsVyrtb4%3D&md5=a31b9c50931ab9eaa149a41705423c07CAS | 21470802PubMed |
Jepsen, R. J., Evans, L. E., and Youngs, C. R. (2010). Use of direct thaw insemination to establish pregnancies with frozen–thawed semen from a standard jack. J. Equine Vet. Sci. 30, 651–656.
| Use of direct thaw insemination to establish pregnancies with frozen–thawed semen from a standard jack.Crossref | GoogleScholarGoogle Scholar |
Kenney, R., Bergman, R., Cooper, W., and Morse, G. (1975). Minimal contamination techniques for breeding mares: techniques and preliminary findings. In ‘Proceedings of the 21st Annual Convention of the American Association of Equine Practitioners, Boston, 1–3 December 1975’. (Ed(s) American Association of Equine Practitioners – AAEP.) pp. 327–336. (American Association of Equine Practitioners – AAEP: Boston, MA.)
Macías García, B., Ortega Ferrusola, C., Aparicio, I. M., Miró-Morán, A., Morillo Rodriguez, A., Gallardo Bolaños, J. M., González Fernández, L., Balao da Silva, C. M., Rodríguez Martínez, H., Tapia, J. A., and Peña, F. J. (2012). Toxicity of glycerol for the stallion spermatozoa: effects on membrane integrity and cytoskeleton, lipid peroxidation and mitochondrial membrane potential. Theriogenology 77, 1280–1289.
| Toxicity of glycerol for the stallion spermatozoa: effects on membrane integrity and cytoskeleton, lipid peroxidation and mitochondrial membrane potential.Crossref | GoogleScholarGoogle Scholar | 22326587PubMed |
Madison, R. J., Evans, L. E., and Youngs, C. R. (2013). The effect of 2-hydroxypropyl-β-cyclodextrin on post-thaw parameters of cryopreserved jack and stallion semen. J. Equine Vet. Sci. 33, 272–278.
| The effect of 2-hydroxypropyl-β-cyclodextrin on post-thaw parameters of cryopreserved jack and stallion semen.Crossref | GoogleScholarGoogle Scholar |
Mantovani, R., Rota, A., Falomo, M. E., Bailoni, L., and Vincenti, L. (2002). Comparison between glycerol and ethylene glycol for the cryopreservation of equine spermatozoa: semen quality assessment with standard analyses and with the hypo-osmotic swelling test. Reprod. Nutr. Dev. 42, 217–226.
| Comparison between glycerol and ethylene glycol for the cryopreservation of equine spermatozoa: semen quality assessment with standard analyses and with the hypo-osmotic swelling test.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XovVWns7g%3D&md5=b5578c8ab7acd5426251e4effd41bf13CAS | 12405450PubMed |
Martins-Bessa, A., Rocha, A., and Mayenco-Aguirre, A. (2006). Comparing ethylene glycol with glycerol for cryopreservation of canine semen in egg-yolk TRIS extenders. Theriogenology 66, 2047–2055.
| Comparing ethylene glycol with glycerol for cryopreservation of canine semen in egg-yolk TRIS extenders.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28Xht1SmsLnO&md5=f3adee52e6122794129e71678cb57435CAS | 16879862PubMed |
Medeiros, A., Gomes, G., Carmo, M., Papa, F., and Alvarenga, M. (2002). Cryopreservation of stallion spermatoza using different amides. Theriogenology 58, 273–276.
| 1:CAS:528:DC%2BD38XkvFynuro%3D&md5=0270ac94c67436796b898e7f7e9ab80eCAS |
Miró, J., Lobo, V., Quintero-Moreno, A., Medrano, A., Peña, A., and Rigau, T. (2005). Sperm motility patterns and metabolism in Catalonian donkey semen. Theriogenology 63, 1706–1716.
| Sperm motility patterns and metabolism in Catalonian donkey semen.Crossref | GoogleScholarGoogle Scholar | 15763113PubMed |
Miró, J., Taberner, E., Rivera, M., Peña, A., Medrano, A., Rigau, T., and Peñalba, A. (2009). Effects of dilution and centrifugation on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey semen. Theriogenology 72, 1017–1022.
| Effects of dilution and centrifugation on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey semen.Crossref | GoogleScholarGoogle Scholar | 19747718PubMed |
Miró, J., Vilés, K., Fernández, M., Fábregas, N., Soares, J., and García, W. (2011). Induced acute endometritis by frozen semen insemination in donkey. Reprod. Domest. Anim. 46, 130.
Molinia, F. C., Evans, G., and Maxwell, W. M. C. (1994). Incorporation of penetrating cryoprotectants in diluents for pellet-freezing ram spermatozoa. Theriogenology 42, 849–858.
| Incorporation of penetrating cryoprotectants in diluents for pellet-freezing ram spermatozoa.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2cXmvFKnurs%3D&md5=62103e5fc587c1924965b2aebd35bf1eCAS | 16727590PubMed |
Moore, A. I., Squires, E. L., Bruemmer, J. E., and Graham, J. K. (2006). Effect of cooling rate and cryoprotectant on the cryosurvival of equine spermatozoa. J. Equine Vet. Sci. 26, 215–218.
| Effect of cooling rate and cryoprotectant on the cryosurvival of equine spermatozoa.Crossref | GoogleScholarGoogle Scholar |
Morillo Rodríguez, A., Ortega Ferrusola, C., Macías García, B., Morrell, J. M., Rodríguez Martínez, H., Tapia, J. A., and Peña, F. J. (2011). Freezing stallion semen with the new Cáceres extender improves post-thaw sperm quality and diminishes stallion-to-stallion variability. Anim. Reprod. Sci. 127, 78.
| Freezing stallion semen with the new Cáceres extender improves post-thaw sperm quality and diminishes stallion-to-stallion variability.Crossref | GoogleScholarGoogle Scholar |
Oldenhof, H., Friedel, K., Sieme, H., Glasmacher, B., and Wolkers, W. F. (2010). Membrane permeability parameters for freezing of stallion spermatozoa as determined by Fourier transform infrared spectroscopy. Cryobiology 61, 115–122.
| Membrane permeability parameters for freezing of stallion spermatozoa as determined by Fourier transform infrared spectroscopy.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXpvVGktr0%3D&md5=30de797246cef7400e3d0cf056a799c0CAS | 20553897PubMed |
Oliveira, J. V., Alvarenga, M. A., Melo, C. M., Macedo, L. M., DellÁqua, J. A., and Papa, F. O. (2006). Effect of cryoprotectant on donkey semen freezability and fertility. Anim. Reprod. Sci. 94, 82–84.
Ortiz, I., Dorado, J., Acha, D., Gálvez, M. J., Urbano, M., and Hidalgo, M. (2015). Colloid single-layer centrifugation improves post-thaw donkey (Equus asinus) sperm quality and is related to ejaculate freezability. Reprod. Fertil. Dev. 27, 332–340.
| Colloid single-layer centrifugation improves post-thaw donkey (Equus asinus) sperm quality and is related to ejaculate freezability.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC2MzlvFCmtA%3D%3D&md5=e7a033cc979ced7c60f0a23ed4337c72CAS | 25482321PubMed |
Peña, F. J., Macías García, B., Samper, J. C., Aparicio, I. M., Tapia, J. A., and Ortega Ferrusola, C. (2011). Dissecting the molecular damage to stallion spermatozoa: the way to improve current cryopreservation protocols? Theriogenology 76, 1177–1186.
| Dissecting the molecular damage to stallion spermatozoa: the way to improve current cryopreservation protocols?Crossref | GoogleScholarGoogle Scholar | 21835453PubMed |
Pillet, E., Batellier, F., Duchamp, G., Furstoss, V., Le Vern, Y., Kerboeuf, D., Desherces, S., Schmitt, E., and Magistrini, M. (2008). High fertility rates with stallion spermatozoa cryopreserved in INRA96-based extender are not predicted by in vitro parameters. Anim. Reprod. Sci. 107, 339–340.
| High fertility rates with stallion spermatozoa cryopreserved in INRA96-based extender are not predicted by in vitro parameters.Crossref | GoogleScholarGoogle Scholar |
Rota, A., Panzani, D., Sabatini, C., and Camillo, F. (2012). Donkey jack (Equus asinus) semen cryopreservation: studies of seminal parameters, post breeding inflammatory response and fertility in donkey jennies. Theriogenology 78, 1846–1854.
| Donkey jack (Equus asinus) semen cryopreservation: studies of seminal parameters, post breeding inflammatory response and fertility in donkey jennies.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC38bntFCqsQ%3D%3D&md5=d0fefb195755475d138a64ecbcb8f405CAS | 22979965PubMed |
Serres, C. (2003). Evaluación y Conservación del semen en el asno Zamorano-Leonés. Doctoral Thesis, Universidad Complutense de Madrid.
Serres, C., Álvarez, A., Rodriguez, A., Santiago, I., Mateos, E., and Gómez-Cuétaral, C. (2004a). Effect of extender and glycerol concentration on cryopreservation of semen from the Zamorano–Leonés donkey. Reprod. Domest. Anim. 39, 267.
Serres, C., Alvarez, A. L., Rodriguez, A., Crespo, F., Mateos, E., and Gomez-Cuetara, C. (2004b). Use of ethylene glycol in the cryopreservation of Zamorano–Leones donkey semen. In ‘Proceedings of the 14th International Congress on Animal Reproduction, Porto Seguro, 8–11 August 2000’. (Ed(s) M. Henry, P.K. Basrur, P.J. Broadbent and L.E. Pinheiro.) p. 73. (International Congress on Animal Reproduction – ICAR: Porto Seguro, Brazil.)
Serres, C., Gutiérrez-Cepeda, L., Gero, S., Gómez, V., Blázquez, J. C., and Crespo, F. (2014). Fertility rates in jennies by new semen-processing techniques. J. Equine Vet. Sci. 34, 193–194.
| Fertility rates in jennies by new semen-processing techniques.Crossref | GoogleScholarGoogle Scholar |
Smits, K., Hoogewijs, M., Woelders, H., Daels, P., and Van Soom, A. (2012). Breeding or assisted reproduction? Relevance of the horse model applied to the conservation of endangered equids. Reprod. Domest. Anim. 47, 239–248.
| Breeding or assisted reproduction? Relevance of the horse model applied to the conservation of endangered equids.Crossref | GoogleScholarGoogle Scholar | 22827377PubMed |
Snoeck, P., Cottorello, A., and Henry, M. (2012). Viability and fertility of stallion semen frozen with ethylene glycol and acetamide as a cryogenic agent. Anim. Reprod. 9, 33–39.
Squires, E. L., Keith, S. L., and Graham, J. K. (2004). Evaluation of alternative cryoprotectants for preserving stallion spermatozoa. Theriogenology 62, 1056–1065.
| Evaluation of alternative cryoprotectants for preserving stallion spermatozoa.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXmt1Knu7k%3D&md5=71ebc203f8d15a91bb89c705b9d2fca3CAS | 15289047PubMed |
Trimeche, A., Renard, P., and Tainturier, D. (1998). A procedure for Poitou jackass sperm cryopreservation. Theriogenology 50, 793–806.
| A procedure for Poitou jackass sperm cryopreservation.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3c7pvVKhug%3D%3D&md5=09450dce78d07c494e48889922ad1004CAS | 10734453PubMed |
Vidament, M., Daire, C., Yvon, J. M., Doligez, P., Bruneau, B., Magistrini, M., and Ecot, P. (2002). Motility and fertility of stallion semen frozen with glycerol and/or dimethyl formamide. Theriogenology 58, 249–251.
| 1:CAS:528:DC%2BD38XkvFynur4%3D&md5=729c27c56bec322de974395de5d14c71CAS |
Vidament, M., Vincent, P., Yvon, J., Bruneau, B., and Martin, F. (2005). Glycerol in semen extender is a limiting factor in the fertility in asine and equine species. Anim. Reprod. Sci. 89, 302.
| 1:STN:280:DC%2BD2MrotF2rug%3D%3D&md5=1430625745a56bf859cb1abeea7543ffCAS | 16265750PubMed |
Vidament, M., Vincent, P., Martin, F., Magistrini, M., and Blesbois, E. (2009). Differences in ability of jennies and mares to conceive with cooled and frozen semen containing glycerol or not. Anim. Reprod. Sci. 112, 22–35.
| Differences in ability of jennies and mares to conceive with cooled and frozen semen containing glycerol or not.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXivVWktbw%3D&md5=c05f5c5c15e9ca74be5db50fc0b8c96fCAS | 18502059PubMed |
Vilés, K., Rabanal, R., Rodríguez-Prado, M., and Miró, J. (2013). Effect of ketoprofen treatment on the uterine inflammatory response after AI of jennies with frozen semen. Theriogenology 79, 1019–1026.
| Effect of ketoprofen treatment on the uterine inflammatory response after AI of jennies with frozen semen.Crossref | GoogleScholarGoogle Scholar | 23453786PubMed |
Watson, P. F. (1995). Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reprod. Fertil. Dev. 7, 871–891.
| Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK283ns1alsQ%3D%3D&md5=368343d88edba9b5012d4dc3341aa1e0CAS | 8711221PubMed |
