Evaluación de inductores de la capacitación espermática en semen criopreservado de asnos (equus asinus)
Las biotecnologías reproductivas, constituyen una serie de herramientas para la conservación de las razas consideradas en peligro de extinción como los burros. El objetivo de esta investigación fue evaluar el uso de inductores de capacitación espermática en semen congelado-descongelado de asnos. Se...
- Autores:
-
Mendoza Arenas, Luis Mendoza
- Tipo de recurso:
- Fecha de publicación:
- 2022
- Institución:
- Universidad Nacional de Colombia
- Repositorio:
- Universidad Nacional de Colombia
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.unal.edu.co:unal/83291
- Palabra clave:
- 590 - Animales
Conservación de semen
Animales - Conducta sexual
Pentoxifilina
Biotecnología
Biotechnology
Pentoxifylline
- Rights
- openAccess
- License
- Reconocimiento 4.0 Internacional
| id |
UNACIONAL2_08c796e81c68dae650f3a47bda4f1ee6 |
|---|---|
| oai_identifier_str |
oai:repositorio.unal.edu.co:unal/83291 |
| network_acronym_str |
UNACIONAL2 |
| network_name_str |
Universidad Nacional de Colombia |
| repository_id_str |
|
| dc.title.spa.fl_str_mv |
Evaluación de inductores de la capacitación espermática en semen criopreservado de asnos (equus asinus) |
| dc.title.translated.eng.fl_str_mv |
Evaluation of spermatic capacity inducers in criopreserved semen of donkeys (Equus asinus) |
| title |
Evaluación de inductores de la capacitación espermática en semen criopreservado de asnos (equus asinus) |
| spellingShingle |
Evaluación de inductores de la capacitación espermática en semen criopreservado de asnos (equus asinus) 590 - Animales Conservación de semen Animales - Conducta sexual Pentoxifilina Biotecnología Biotechnology Pentoxifylline |
| title_short |
Evaluación de inductores de la capacitación espermática en semen criopreservado de asnos (equus asinus) |
| title_full |
Evaluación de inductores de la capacitación espermática en semen criopreservado de asnos (equus asinus) |
| title_fullStr |
Evaluación de inductores de la capacitación espermática en semen criopreservado de asnos (equus asinus) |
| title_full_unstemmed |
Evaluación de inductores de la capacitación espermática en semen criopreservado de asnos (equus asinus) |
| title_sort |
Evaluación de inductores de la capacitación espermática en semen criopreservado de asnos (equus asinus) |
| dc.creator.fl_str_mv |
Mendoza Arenas, Luis Mendoza |
| dc.contributor.advisor.none.fl_str_mv |
Restrepo Betancur, Giovanni |
| dc.contributor.author.none.fl_str_mv |
Mendoza Arenas, Luis Mendoza |
| dc.contributor.researchgroup.spa.fl_str_mv |
GIBA (Grupo de Investigación Biotecnología Animal) |
| dc.subject.ddc.spa.fl_str_mv |
590 - Animales |
| topic |
590 - Animales Conservación de semen Animales - Conducta sexual Pentoxifilina Biotecnología Biotechnology Pentoxifylline |
| dc.subject.lemb.none.fl_str_mv |
Conservación de semen Animales - Conducta sexual |
| dc.subject.proposal.spa.fl_str_mv |
Pentoxifilina Biotecnología |
| dc.subject.proposal.eng.fl_str_mv |
Biotechnology Pentoxifylline |
| description |
Las biotecnologías reproductivas, constituyen una serie de herramientas para la conservación de las razas consideradas en peligro de extinción como los burros. El objetivo de esta investigación fue evaluar el uso de inductores de capacitación espermática en semen congelado-descongelado de asnos. Se colectaron 5 asnos, localizados en el Valle de Aburrá con edades entre 4-8 años, se obtuvieron 3 eyaculados por animal, mediante vagina artificial, el semen recién recolectado fue diluido en proporción 1:1 en Equiplus® para ser criopreservado. Después, se descongelaron dos pajillas de 0.5 mL cada una del mismo eyaculado en un baño María a 37 °C, este volumen fue divido en 8 submuetras y 125 μL de semen fueron depositados en tubos eppendorf, posteriormente se adicionó a cada tubo el inductor de capacitación pentoxifilina (PTX) 2.5, 5 y 7.5 mM, ácido etilendiaminotetraacético (EDTA) 0.5, 1 y 1.5 mM, heparina (HEP) 1.1 mM como control positivo y sin aditivo (control) por 20 minutos. Se evaluaron variables de cinética espermática sistema IVOS (Hamilton Thorne), vitalidad espermática (VE) mediante el kit de viabilidad LIVE/DEAD, integridad acrosómica con (FITC-PSA), mientras que la capacitación, potencial mitocondrial e ingreso de Ca2+, fue evaluado usando un citómetro de flujo (FORTESSA LSR, BD Biosciences, EE. UU). Se observó que la movilidad total (MT) y la vitalidad espermática (VE) aumentaron en relación con el control, con el uso de 2,5 y 5.0 mM de PTX respectivamente (p<0.05), mientras que la movilidad progresiva (MP) disminuyó para las 3 concentraciones de EDTA (p<0.05), otras variables como velocidad lineal (VSL), velocidad curvilínea (VCL), amplitud lateral de la cabeza (ALH) y frecuencia de batida (BCF) disminuyen su proporción (p<0.05) al adicionar de EDTA 1, 1.5 y PTX 7.5 mM. En cuanto a la capacitación, se encontró que las concentraciones de EDTA 1, 1.5 mM y PTX 5, 7.5 mM (p<0.01) favorecen eventos asociados a la capacitación espermática y además disminuye la concentración de Ca2+ intracelular para todas las concentraciones de PTX y EDTA 0.5 mM (p<0.01). No se observaron efectos en la actividad mitocondrial. Se concluye que la adición de PTX a 2.5, 5 mM, incrementa la proporción de la MT y VE, por el contrario, la MP disminuye al aumentar la concentración de EDTA, además cambios en la estructura de la membrana plasmática asociados a eventos de capacitación de semen congelado-descongelado de burro, puede ser modulada por la suplementación con PTX (5 mM y 7.5 mM) y EDTA (1 mM y 1.5 mM). Sin embargo, la adición de estos inductores de la capacitación, no logró influencia en el ingreso de Ca2+, por lo tanto, se requiere mayor nivel de investigación ya que los mecanismos e implicaciones en la capacitación espermática para semen de burro aún son desconocidos. (Tomado de la fuente) |
| publishDate |
2022 |
| dc.date.issued.none.fl_str_mv |
2022-02-02 |
| dc.date.accessioned.none.fl_str_mv |
2023-02-03T18:35:52Z |
| dc.date.available.none.fl_str_mv |
2023-02-03T18:35:52Z |
| dc.type.spa.fl_str_mv |
Trabajo de grado - Maestría |
| dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/masterThesis |
| dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/acceptedVersion |
| dc.type.content.spa.fl_str_mv |
Text |
| dc.type.redcol.spa.fl_str_mv |
http://purl.org/redcol/resource_type/TM |
| status_str |
acceptedVersion |
| dc.identifier.uri.none.fl_str_mv |
https://repositorio.unal.edu.co/handle/unal/83291 |
| dc.identifier.instname.spa.fl_str_mv |
Universidad Nacional de Colombia |
| dc.identifier.reponame.spa.fl_str_mv |
Repositorio Institucional Universidad Nacional de Colombia |
| dc.identifier.repourl.spa.fl_str_mv |
https://repositorio.unal.edu.co/ |
| url |
https://repositorio.unal.edu.co/handle/unal/83291 https://repositorio.unal.edu.co/ |
| identifier_str_mv |
Universidad Nacional de Colombia Repositorio Institucional Universidad Nacional de Colombia |
| dc.language.iso.spa.fl_str_mv |
spa |
| language |
spa |
| dc.relation.indexed.spa.fl_str_mv |
LaReferencia |
| dc.relation.references.spa.fl_str_mv |
Acha, D., Hidalgo, M., Ortiz, I., Gálvez, M. J., Carrasco, J. J., Gómez-Arrones, V., & Dorado, J. (2016). Freezability of Andalusian donkey (Equus asinus) spermatozoa: Effect of extenders and permeating cryoprotectants. Reproduction, Fertility and Development, 28(12), 1990–1998. https://doi.org/10.1071/RD14449 Arenas, E., Cambron, A., Ambriz, D., Zuñiga, P., Rodriguez, A., & Rosado, A. (2010). Bases fisiologicas de la capacitacion y de la reaccion acrosomal del espermatozoide. Contactos, 78, 5–11. Ball, G. F., & Balthazart, J. (2008). How useful is the appetitive and consummatory distinction for our understanding of the neuroendocrine control of sexual behavior? In Hormones and Behavior (Vol. 53, Issue 2, pp. 307–311). NIH Public Access. https://doi.org/10.1016/j.yhbeh.2007.09.023 Barakat, I. A. H., Danfour, M. A., Galewan, F. A. M., & Dkhil, M. A. (2015). Effect of Various Concentrations of Caffeine, Pentoxifylline, and Kallikrein on Hyperactivation of Frozen Bovine Semen. BioMed Research International, 2015, 948575. https://doi.org/10.1155/2015/948575 Bennett, R., & Pfuderer, S. (2020). The potential for new donkey farming systems to supply the growing demand for hides. Animals, 10(4). https://doi.org/10.3390/ani10040718 Bhattacharya, S. (2018). Cryopretectants and Their Usage in Cryopreservation Process. Cryopreservation Biotechnology in Biomedical and Biological Sciences. https://doi.org/10.5772/intechopen.80477 Blench, R. (2012). Wild Asses and Donkeys in Africa: Interdisciplinary Evidence for Their Biogeography, History and Current Use. Soas, May 2012. Bollwein, H., Fuchs, I., & Koess, C. (2008). Interrelationship Between Plasma Membrane Integrity, Mitochondrial Membrane Potential and DNA Fragmentation in Cryopreserved Bovine Spermatozoa. Reproduction in Domestic Animals, 43(2), 189–195. https://doi.org/10.1111/j.1439-0531.2007.00876.x Boni, R., Gallo, A., & Cecchini, S. (2017). Kinetic activity, membrane mitochondrial potential, lipid peroxidation, intracellular pH and calcium of frozen/thawed bovine spermatozoa treated with metabolic enhancers. Andrology, 5(1), 133–145. https://doi.org/10.1111/andr.12259 Breitbart, H., & Finkelstein, M. (2015). Regulation of Sperm Capacitation and the Acrosome Reaction by PIP2 and Actin Modulation. Asian Journal of Andrology, 17(4), 597–600. https://doi.org/10.4103/1008-682X.154305 Breitbart, H., & Shabtay, O. (2018). Sperm acrosome reaction. In Encyclopedia of Reproduction (Second Edi, Vol. 3). Elsevier. https://doi.org/10.1016/B978-0-12-801238-3.66186-X Brener, E., Rubinstein, S., Cohen, G., Shternall, K., Rivlin, J., & Breitbart, H. (2003). Remodeling of the actin cytoskeleton during mammalian sperm capacitation and acrosome reaction. Biology of Reproduction, 68(3), 837–845. https://doi.org/10.1095/biolreprod.102.009233 Buffone, M. G., Wertheimer, E. V., Visconti, P. E., & Krapf, D. (2014). Central role of soluble adenylyl cyclase and cAMP in sperm physiology. In Biochimica et Biophysica Acta - Molecular Basis of Disease (Vol. 1842, Issue 12, pp. 2610–2620). Elsevier. https://doi.org/10.1016/j.bbadis.2014.07.013 Bustamante, I. C., Pederzolli, C. D., Sgaravatti, A. M., Gregory, R. M., Dutra Filho, C. S., Jobim, M. I. M., & Mattos, R. C. (2009). Skim milk-egg yolk based semen extender compensates for non-enzymatic antioxidant activity loss during equine semen cryopreservation. In Anim. Reprod., v (Vol. 6, Issue 2). Colégio Brasileiro de Reprodução Animal. Calogero, A. E., Fishel, S., Hall, J., Ferrara, E., Vicari, E., Green, S., Hunter, A., Burrello, N., Thornton, S., & D’Agata, R. (1998). Correlation between intracellular cAMP content, kinematic parameters and hyperactivation of human spermatozoa after incubation with pentoxifylline. Human Reproduction, 13(4), 911–915. https://doi.org/10.1093/humrep/13.4.911 Canisso, I. F., Carvalho, G. R., Morel, M. C. G. D., Guimarães, J. D., & McDonnell, S. M. (2010). Sexual behavior and ejaculate characteristics in Pêga donkeys (Equus asinus) mounting estrous horse mares (Equus caballus). Theriogenology, 73(1), 56–63. https://doi.org/10.1016/j.theriogenology.2009.07.026 Canisso, I. F., & McDonnell, S. M. (2010). Donkey breeding behavior with an emphasis on the Pêga breed. MATTHEWS NS &TAYLOR TS.Veterinary Care of Donkeys.Ithaca: International Veterinary Information Service, 310. Canisso, I. F., Panzani, D., Miró, J., & Ellerbrock, R. E. (2019). Key Aspects of Donkey and Mule Reproduction. Veterinary Clinics of North America - Equine Practice, 35(3), 607–642. https://doi.org/10.1016/j.cveq.2019.08.014 Carluccio, A., Contri, A., Amendola, S., De Angelis, E., De Amicis, I., & Mazzatenta, A. (2013). Male isolation: A behavioral representation of the pheromonal “female effect” in donkey (Equus asinus). Physiology and Behavior, 118, 1–7. https://doi.org/10.1016/j.physbeh.2013.04.005 Carminati, D., & Tidona, F. (2017). Nutritional Value and Potential Health Benefits of Donkey Milk. In Nutrients in Dairy and Their Implications for Health and Disease. Elsevier Inc. https://doi.org/10.1016/B978-0-12-809762-5.00031-0 Carneiro, G. F., Cavalcante Lucena, J. E., & de Oliveira Barros, L. (2018). The Current Situation and Trend of the Donkey Industry in South America. Journal of Equine Veterinary Science, 65, 106–110. https://doi.org/10.1016/j.jevs.2018.03.007 Carvalho, L. E., Silva Filho, J. M., Palhares, M. S., Sales, A. L. R., Gonczarowska, A. T., Oliveira, H. N., Resende, M. C., Neves, M. G., & Madison, R. J. (2017). Physical characteristics and fertility of fractionated donkey semen cooled at 5°C. Arquivo Brasileiro de Medicina Veterinaria e Zootecnia, 69(1), 29–38. https://doi.org/10.1590/1678-4162-7976 Consuegra, C., Crespo, F., Bottrel, M., Ortiz, I., Dorado, J., Diaz-Jimenez, M., Pereira, B., & Hidalgo, M. (2018). Stallion sperm freezing with sucrose extenders: A strategy to avoid permeable cryoprotectants. Animal Reproduction Science, 191, 85–91. https://doi.org/10.1016/j.anireprosci.2018.02.013 Contri, A., Gloria, A., Robbe, D., De Amicis, I., & Carluccio, A. (2012a). Characteristics of donkey spermatozoa along the length of the epididymis. Theriogenology, 77(1), 166–173. https://doi.org/10.1016/j.theriogenology.2011.07.031 Contri, A., Gloria, A., Robbe, D., Sfirro, M. P., & Carluccio, A. (2012b). Effect of sperm concentration on characteristics of frozen-thawed semen in donkeys. Animal Reproduction Science, 136(1–2), 74–80. https://doi.org/10.1016/j.anireprosci.2012.10.022 Cortés-Gutiérrez, E. I., Dávila-Rodríguez, M. I., López-Fernández, C., Fernández, J. L., Crespo, F., & Gosálvez, J. (2014). Localization of alkali-labile sites in donkey (Equus asinus) and stallion (Equus caballus) spermatozoa. Theriogenology, 81(2), 321–325. https://doi.org/10.1016/J.THERIOGENOLOGY.2013.10.001 Darwin, C. (1859). Origem das espécies: E a seleção natural. Davoodi, F., Raisi, A., Rajabzadeh, A., Hablolvarid, M. H., & Zakian, A. (2020). The effects of verapamil and heparin co-administration on sperm parameters and oxidative stress in prevention of testicular torsion/detorsion damage in rats. Andrologia, 52(2), 1–8. https://doi.org/10.1111/and.13479 de Andrade, A. F. C., Zaffalon, F. G., Celeghini, E. C. C., Nascimento, J., Tarragó, O. F. B., Martins, S., Alonso, M. A., & Arruda, R. P. (2011). Addition of Seminal Plasma to Post-thawing Equine Semen: What is the Effect on Sperm Cell Viability? Reproduction in Domestic Animals, 46(4), 682–686. https://doi.org/https://doi.org/10.1111/j.1439-0531.2010.01729.x de Vasconcelos Franco, J. S., Faheem, M., Chaveiro, A., & Moreira da Silva, F. (2016). Effects of α-tocopherol and freezing rates on the quality and heterologous in vitro fertilization capacity of stallion sperm after cryopreservation. Theriogenology, 86(4), 957–962. https://doi.org/10.1016/j.theriogenology.2016.03.019 Demyda-Peyrás, S., Bottrel, M., Acha, D., Ortiz, I., Hidalgo, M., Carrasco, J. J., Gómez-Arrones, V., Gósalvez, J., & Dorado, J. (2018). Effect of cooling rate on sperm quality of cryopreserved Andalusian donkey spermatozoa. Animal Reproduction Science, 193, 201–208. https://doi.org/10.1016/j.anireprosci.2018.04.069 Dhanju, C. K., Toor, G., & Cheema, R. S. (2009). Effect of seminal plasma on quantily and qualily of proteins leaked during heparin - induced in vitro capacitation of bull sperm. Indian Journal of Animal Research, 43(2), 136–138. Di Palma, T., Cecchini, S., MacChia, G., Pasolini, M. P., Cocchia, N., & Boni, R. (2020). Kinematic, bioenergetic and oxidative evaluations of donkey sperm preserved at +4°C. Zygote, 1–8. https://doi.org/10.1017/S096719942000012X Di Paolo, G., & De Camilli, P. (2006). Phosphoinositides in cell regulation and membrane dynamics. Nature, 443(7112), 651–657. https://doi.org/10.1038/nature05185 Di Santo, M., Tarozzi, N., Nadalini, M., & Borini, A. (2012). Human sperm cryopreservation: Update on techniques, effect on DNA integrity, and implications for ART. Advances in Urology, 2012. https://doi.org/10.1155/2012/854837 Diaz-Jimenez, M., Dorado, J., Ortiz, I., Consuegra, C., Pereira, B., Gonzalez-De Cara, C. A., Aguilera, R., Mari, G., Mislei, B., Love, C. C., & Hidalgo, M. (2018). Cryopreservation of donkey sperm using non-permeable cryoprotectants. Animal Reproduction Science, 189(December 2017), 103–109. https://doi.org/10.1016/j.anireprosci.2017.12.013 Dong, W. W., Zeng, Z. Da, Au, D., Chan, C. O., Yang, D. J., & Li, X. B. (2012). Quality control of Colla corii asini using near-infrared spectroscopy and chemometrics clustering techniques. Journal of Food and Drug Analysis, 20(1), 152–158. Dorado, J., Acha, D., Ortiz, I., Gálvez, M. J., Carrasco, J. J., Díaz, B., Gómez-Arrones, V., Calero-Carretero, R., & Hidalgo, M. (2013). Relationship between conventional semen characteristics, sperm motility patterns and fertility of Andalusian donkeys (Equus asinus). Animal Reproduction Science, 143(1–4), 64–71. https://doi.org/10.1016/j.anireprosci.2013.10.003 Dorado, J., Hidalgo, M., Acha, D., Ortiz, I., Bottrel, M., Azcona, F., Carrasco, J. J., Gómez-Arrones, V., & Demyda-Peyrás, S. (2019). Cryopreservation of Andalusian donkey (Equus asinus) spermatozoa: Use of alternative energy sources in the freezing extender affects post-thaw sperm motility patterns but not DNA stability. Animal Reproduction Science, 208(June), 106126. https://doi.org/10.1016/j.anireprosci.2019.106126 Douet, C., Reigner, F., Barrière, P., Blard, T., Deleuze, S., & Goudet, G. (2019). First attempts for vitrification of immature oocytes in donkey (Equus asinus): Comparison of two vitrification methods. Theriogenology, 126, 261–265. https://doi.org/10.1016/j.theriogenology.2018.12.030 El-Shahat, K. H., Taysser, M. I., Badr, M. R., & Zaki, K. A. (2016). Effect of heparin, caffeine and calcium ionophore A23187 on in vitro induction of the acrosome reaction of fresh ram spermatozoa. Asian Pacific Journal of Reproduction, 5(2), 148–155. https://doi.org/10.1016/j.apjr.2016.01.012 Esteves, S. C., Spaine, D. M., & Cedenho, A. P. (2007). Effects of pentoxifylline treatment before freezing on motility, viability and acrosome status of poor quality human spermatozoa cryopreserved by the liquid nitrogen vapor method. Brazilian Journal of Medical and Biological Research, 40(7), 985–992. https://doi.org/10.1590/S0100-879X2006005000118 Fernández, S., Morado, S., Cetica, P., & Córdoba, M. (2020). Hyaluronic acid capacitation induces intracellular signals modulated by membrane-associated adenylate cyclase and tyrosine kinase involved in bovine in vitro fertilization. Theriogenology, 148, 174–179. https://doi.org/10.1016/j.theriogenology.2020.02.033 Ferrante, A., Baca Castex, C., Bruno, S., Arraztoa, C., Plaza, J., Neild, D., & Miragaya, M. (2018a). Comparison of Whole and Centrifuged Egg Yolk Added to Kenney’s and Lactose-EDTA Extenders for Donkey Semen Cryopreservation. Journal of Equine Veterinary Science, 65, 12–18. https://doi.org/10.1016/j.jevs.2018.02.024 Ferrante, A., Baca Castex, C., Bruno, S., Arraztoa, C., Plaza, J., Neild, D., & Miragaya, M. (2018b). Comparison of Whole and Centrifuged Egg Yolk Added to Kenney’s and Lactose-EDTA Extenders for Donkey Semen Cryopreservation. Journal of Equine Veterinary Science, 65, 12–18. https://doi.org/10.1016/J.JEVS.2018.02.024 Ferré, L. B., Kjelland, M. E., Strøbech, L. B., Hyttel, P., Mermillod, P., & Ross, P. J. (2020). Review: Recent advances in bovine in vitro embryo production: reproductive biotechnology history and methods. Animal, 14(5), 991–1004. https://doi.org/10.1017/S1751731119002775 Finkelstein, M., Etkovitz, N., & Breitbart, H. (2010). Role and regulation of sperm gelsolin prior to fertilization. The Journal of Biological Chemistry, 285(51), 39702–39709. https://doi.org/10.1074/jbc.M110.170951 Finkelstein, M., Megnagi, B., Ickowicz, D., & Breitbart, H. (2013). Regulation of sperm motility by PIP2(4,5) and actin polymerization. Developmental Biology, 381(1), 62–72. https://doi.org/10.1016/j.ydbio.2013.06.014 Fredholm, B. B., & Lindström, K. (1986). The Xanthine Derivative 1-(5′-Oxohexyl)-3-methyl-7-propyl Xanthine (HWA 285) Enhances the Actions of Adenosine. Acta Pharmacologica et Toxicologica, 58(3), 187–192. https://doi.org/https://doi.org/10.1111/j.1600-0773.1986.tb00093.x Gamboa, S., Rodrigues, A. S., Henriques, L., Batista, C., & Ramalho-Santos, J. (2010). Seasonal functional relevance of sperm characteristics in equine spermatozoa. Theriogenology, 73(7), 950–958. https://doi.org/10.1016/j.theriogenology.2009.11.023 García-Álvarez, O., Maroto-Morales, A., Jiménez-Rabadán, P., Ramón, M., del Olmo, E., Iniesta-Cuerda, M., Anel-López, L., Fernández-Santos, M. R., Garde, J. J., & Soler, A. J. (2015). Effect of different media additives on capacitation of frozen–thawed ram spermatozoa as a potential replacement for estrous sheep serum. Theriogenology, 84(6), 948–955. https://doi.org/10.1016/J.THERIOGENOLOGY.2015.05.032 Gervasi, M. G., Marczylo, T. H., Lam, P. M., Rana, S., Franchi, A. M., Konje, J. C., & Perez-Martinez, S. (2013). Anandamide Levels Fluctuate in the Bovine Oviduct during the Oestrous Cycle. PLoS ONE, 8(8), e72521. https://doi.org/10.1371/journal.pone.0072521 Gervasi, M. G., & Visconti, P. E. (2016). Chang’s meaning of capacitation: A molecular perspective. Molecular Reproduction and Development, 83(10), 860–874. https://doi.org/10.1002/mrd.22663 Ghasemzadeh, A., Karkon-Shayan, F., Yousefzadeh, S., Naghavi-Behzad, M., & Hamdi, K. (2016). Study of pentoxifylline effects on motility and viability of spermatozoa from infertile asthenozoospermic males. Nigerian Medical Journal, 57(6), 324. https://doi.org/10.4103/0300-1652.193857 Glogowski, J., Danforth, D. R., & Ciereszko, A. (2002). Inhibition of alkaline phosphatase activity of boar semen by pentoxifylline, caffeine, and theophylline. Journal of Andrology, 23(6), 783–792. Gloria, A., Contri, A., De Amicis, I., Robbe, D., & Carluccio, A. (2011). Differences between epididymal and ejaculated sperm characteristics in donkey. Animal Reproduction Science, 128(1–4), 117–122. https://doi.org/10.1016/j.anireprosci.2011.09.007 Grötter, L. G., Cattaneo, L., Marini, P. E., Kjelland, M. E., & Ferré, L. B. (2019). Recent advances in bovine sperm cryopreservation techniques with a focus on sperm post-thaw quality optimization. Reproduction in Domestic Animals, 54(4), 655–665. https://doi.org/10.1111/rda.13409 Guasti, P. N., Monteiro, G. A., Maziero, R. R. D., Carmo, M. T., Dell’Aqua, J. A., Crespilho, A. M., Rifai, E. A., & Papa, F. O. (2017). Pentoxifylline effects on capacitation and fertility of stallion epididymal sperm. Animal Reproduction Science, 179, 27–34. https://doi.org/10.1016/j.anireprosci.2017.01.013 Guo, S., Savolainen, P., Su, J., Zhang, Q., Qi, D., Zhou, J., Zhong, Y., Zhao, X., & Liu, J. (2006). Origin of mitochondrial DNA diversity of domestic yaks. BMC Evolutionary Biology, 6, 73. https://doi.org/10.1186/1471-2148-6-73 Haddad, R., Khan, R., Takahashi, Y. K., Mori, K., Harel, D., & Sobel, N. (2008). A metric for odorant comparison. Nature Methods, 5(5), 425–429. https://doi.org/10.1038/nmeth.1197 Henry, M., Snoeck, P. P. N., & Cottorello, A. C. P. (2002). Post-thaw spermatozoa plasma membrane integrity and motility of stallion semen frozen with different cryoprotectants. Theriogenology, 58(2–4), 245–248. https://doi.org/10.1016/S0093-691X(02)00750-1 Hu, J., Zhong, C., Ding, C., Chi, Q., Walz, A., Mombaerts, P., Matsunami, H., & Luo, M. (2007). Detection of near-atmospheric concentrations of CO2 by an olfactory subsystem in the mouse. Science, 317(5840), 953–957. https://doi.org/10.1126/science.1144233 Hussain, N., Andrabi, S. M. H., & Mehmood, M. U. (2019). Effect of EDTA as Chelating Agent in Extender on the Post Thaw Quality of Buffalo Bull Spermatozoa. Cryo Letters, 40(3), 159–163. Itach, S. B.-S., Finklestein, M., Etkovitz, N., & Breitbart, H. (2012). Hyper-activated motility in sperm capacitation is mediated by phospholipase D-dependent actin polymerization. Developmental Biology, 362(2), 154–161. https://doi.org/10.1016/j.ydbio.2011.12.002 Kaneko, T., & Nakagata, N. (2006). Improvement in the long-term stability of freeze-dried mouse spermatozoa by adding of a chelating agent. Cryobiology, 53(2), 279–282. https://doi.org/10.1016/J.CRYOBIOL.2006.06.004 Keshtgar, S., Iravanpour, F., Gharesi-Fard, B., & Kazerooni, M. (2016). Combined effect of trolox and EDTA on frozen-thawed sperm quality. Iranian Journal of Medical Sciences, 41(3), 230–237. Keverne, E. B. (2002). Pheromones, vomeronasal function, and gender-specific behavior. In Cell (Vol. 108, Issue 6, pp. 735–738). Cell Press. https://doi.org/10.1016/S0092-8674(02)00687-6 Kugler, W., Grunenfelder, H. P., & Broxham, E. (2008). Donkey Breeds in Europe: Inventory, Description, Need for Action, Conservation. In … and Seeds in Europe. Kumeta, Y., Maruyama, T., Asama, H., Yamamoto, Y., Hakamatsuka, T., & Goda, Y. (2014). Species identification of Asini Corii Collas (donkey glue) by PCR amplification of cytochrome b gene. Journal of Natural Medicines, 68(1), 181–185. https://doi.org/10.1007/s11418-013-0790-z Leclerc, P., Sirard, M. A., Chafouleas, J. G., & Lambert, R. D. (1992). Decrease in calmodulin concentrations during heparin-induced capacitation in bovine spermatozoa. Journal of Reproduction and Fertility, 94(1), 23–32. https://doi.org/10.1530/jrf.0.0940023 Li, N., Yu, J., Yang, F., Shao, Y., Wu, S., Liu, B., Li, M., Wang, T., Li, J., & Zeng, S. (2021). L-Proline: An Effective Agent for Frozen and Post-thawed Donkey Semen Storage. Journal of Equine Veterinary Science, 101, 103393. https://doi.org/10.1016/j.jevs.2021.103393 López, L. F. M. (2018). El puente de Occidente y la inegración de Antioquia. Universidad Nacional de Colombia-Sede Medellín. Mackenna, A. (1995). Contribution of the male factor to unexplained infertility: a review. International Journal of Andrology, 18 Suppl 1, 58–61. https://doi.org/10.1111/j.1365-2605.1995.tb00640.x Marshall, F. (2007). African pastoral perspectives on domestication of the donkey. In In Rethinking agriculture: archaeological and ethnoarchaeological perspectives (eds T. P. Denham, J. Iriarte & L. Vrydaghs) (Walnut Cre, pp. 371–407). Martínez-León, E., Osycka-Salut, C., Signorelli, J., Pozo, P., Pérez, B., Kong, M., Morales, P., Pérez-Martínez, S., & Díaz, E. S. (2015). Fibronectin stimulates human sperm capacitation through the cyclic AMP/protein kinase A pathway. Human Reproduction, 30(9), 2138–2151. https://doi.org/10.1093/humrep/dev154 Massip, A. (2001). Cryopreservation of embryos of farm animals. In Reproduction in Domestic Animals (Vol. 36, Issue 2, pp. 49–55). John Wiley & Sons, Ltd. https://doi.org/10.1046/j.1439-0531.2001.00248.x Mazur, P., Leibo, S. P., & Chu, E. H. Y. (1972). A two-factor hypothesis of freezing injury. Evidence from Chinese hamster tissue-culture cells. Experimental Cell Research, 71(2), 345–355. https://doi.org/10.1016/0014-4827(72)90303-5 McDonnell, S. (2003). A Practical Field Guide to Horse Behavior: The Equid Ethogram. Journal of Equine Veterinary Science, 23(1), A1. https://doi.org/10.1016/s0737-0806(03)70087-2 McLean, A. K., Navas González, F. J., & Canisso, I. F. (2019). Donkey and Mule Behavior. Veterinary Clinics of North America - Equine Practice, 35(3), 575–588. https://doi.org/10.1016/j.cveq.2019.08.010 Medeiros, A. S. L., Gomes, G. M., Carmo, M. T., Papa, F. O., & Alvarenga, M. A. (2002). Cryopreservation of stallion sperm using different amides. Theriogenology, 58(2–4), 273–276. https://doi.org/10.1016/S0093-691X(02)00898-1 Mendoza, C., Carreras, 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. Journal of Reproduction and Fertility, 95(3), 755–763. https://doi.org/10.1530/jrf.0.0950755 Mercati, F., Domingo, P., Pasquariello, R., Dall’Aglio, C., Di Michele, A., Forti, K., Cocci, P., Boiti, C., Gil, L., Zerani, M., & Maranesi, M. (2020). Effect of chelating and antioxidant agents on morphology and DNA methylation in freeze-drying rabbit (Oryctolagus cuniculus) spermatozoa. Reproduction in Domestic Animals, 55(1), 29–37. https://doi.org/10.1111/rda.13577 Mesa, A. M., Sc, M., R, G. H., & Sc, M. (2012). Efecto del colesterol y la dimetilformamida sobre parámetros posdescongelación en espermatozoides de caballos criollos colombianos Effect of cholesterol and dimethiyl-formamide on post-thawing parameters in Colombian creole stallion sperm. 17(1), 2908–2915. Meskini, N., Némoz, G., Okyayuz-Baklouti, I., Lagarde, M., & Prigent, A. F. (1994). Phosphodiesterase inhibitory profile of some related xanthine derivatives pharmacologically active on the peripheral microcirculation. Biochemical Pharmacology, 47(5), 781–788. https://doi.org/10.1016/0006-2952(94)90477-4 Miragaya, M. H., Chaves, M. G., & Agüero, A. (2006). Reproductive biotechnology in South American camelids. Small Ruminant Research, 61(2–3), 299–310. https://doi.org/10.1016/J.SMALLRUMRES.2005.07.017 Miragaya, M. H., Neild, D. M., & Alonso, A. E. (2018). A Review of Reproductive Biology and Biotechnologies in Donkeys. Journal of Equine Veterinary Science, 65, 55–61. https://doi.org/10.1016/j.jevs.2017.12.005 Miró, J., Taberner, E., Rivera, M., Peña, A., Medrano, A., Rigau, T., & 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(8), 1017–1022. https://doi.org/10.1016/j.theriogenology.2009.06.012 Mirshokraei, P., Hassanpour, H., Mehdizadeh, A., & Akhavan Taheri, M. (2011). Pentoxifylline induces capacitation and acrosome reaction and improves quality of motility in canine ejaculated spermatozoa. Research in Veterinary Science, 91(2), 281–284. https://doi.org/10.1016/J.RVSC.2011.01.002 Molina, L. C. P., Luque, G. M., Balestrini, P. A., Marín-Briggiler, C. I., Romarowski, A., & Buffone, M. G. (2018). Molecular basis of human sperm capacitation. Frontiers in Cell and Developmental Biology, 6(July), 1–23. https://doi.org/10.3389/fcell.2018.00072 Montoya, J. D., Rojano, B. A., & Betancur, G. R. (2017). Efecto de la suplementación del diluyente sobre la calidad del semen de asno a la descongelación. Archivos de Zootecnia. Nannou, T. K., Jouhara, H., Trembley, J., & Herrmann, J. (2016). Cryopreservation: Methods, equipment and critical concerns. Refrigeration Science and Technology, 22-25-June, 247–258. https://doi.org/10.18462/IIR.ICCRT.2016.0020 Nassar, A., Mahony, M., Blackmore, P., Morshedi, M., Ozgur, K., & Oehninger, S. (1998). Increase of intracellular calcium is not a cause of pentoxifylline- induced hyperactivated motility or acrosome reaction in human sperm. Fertility and Sterility, 69(4), 748–754. https://doi.org/10.1016/S0015-0282(98)00013-2 Naz, R. K., & Rajesh, P. B. (2004). Role of tyrosine phosphorylation in sperm capacitation/acrosome reaction. Reproductive Biology and Endocrinology, 2, 1–12. https://doi.org/10.1186/1477-7827-2-75 Negri, P., Grechi, E., Tomasi, A., Fabbri, E., & Capuzzo, A. (1996). Effectiveness of pentoxifylline in semen preparation for intrauterine insemination. Human Reproduction, 11(6), 1236–1239. https://doi.org/10.1093/oxfordjournals.humrep.a019363 Nguyen, V. K., Somfai, T., Salamone, D., Thu Huong, V. T., Le Thi Nguyen, H., Huu, Q. X., Hoang, A. T., Phan, H. T., Thi Pham, Y. K., & Pham, L. D. (2021). Optimization of donor cell cycle synchrony, maturation media and embryo culture system for somatic cell nuclear transfer in the critically endangered Vietnamese Ỉ pig. Theriogenology, 166, 21–28. https://doi.org/10.1016/J.THERIOGENOLOGY.2021.02.008 Nouri, H., Towhidi, A., Zhandi, M., & Sadeghi, R. (2013). The Effects of Centrifuged Egg Yolk Used with INRA Plus Soybean Lecithin Extender on Semen Quality to Freeze Miniature Caspian Horse Semen. Journal of Equine Veterinary Science, 33(12), 1050–1053. https://doi.org/https://doi.org/10.1016/j.jevs.2013.03.184 O’Connell, M., McClure, N., & Lewis, S. E. M. (2002). The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Human Reproduction, 17(3), 704–709. https://doi.org/10.1093/humrep/17.3.704 Okazaki, T., Yoshida, S., Teshima, H., & Shimada, M. (2011). The addition of calcium ion chelator, EGTA to thawing solution improves fertilizing ability in frozen-thawed boar sperm. Animal Science Journal = Nihon Chikusan Gakkaiho, 82(3), 412–419. https://doi.org/10.1111/j.1740-0929.2010.00856.x Oliveira, J. V. de, Oliveira, P. V. de L. F., Melo e Oña, C. M., Guasti, P. N., Monteiro, G. A., Sancler da Silva, Y. F. R., Papa, P. de M., Alvarenga, M. A., Dell’Aqua Junior, J. A., & Papa, F. O. (2016). Strategies to improve the fertility of fresh and frozen donkey semen. Theriogenology, 85(7), 1267–1273. https://doi.org/10.1016/j.theriogenology.2015.12.010 Orrego, M. T., Melian, S. I., Montenegro, J., Cimato, A. N., Cisale, H., & Piehl, L. L. (2019). Boar sperm protein tyrosine phosphorylation in the presence of egg yolk soluble and low density lipoprotein fractions during cooling. Theriogenology, 123, 151–158. https://doi.org/10.1016/j.theriogenology.2018.09.031 Ortgies, F., Klewitz, J., Görgens, A., Martinsson, G., & Sieme, H. (2012). Effect of procaine, pentoxifylline and trolox on capacitation and hyperactivation of stallion spermatozoa. Andrologia, 44(SUPPL.1), 130–138. https://doi.org/10.1111/j.1439-0272.2010.01150.x Osycka-Salut, C. E., Castellano, L., Fornes, D., Beltrame, J. S., Alonso, C. A. I., Jawerbaum, A., Franchi, A., Díaz, E. S., & Perez Martinez, S. (2017). Fibronectin From Oviductal Cells Fluctuates During the Estrous Cycle and Contributes to Sperm–Oviduct Interaction in Cattle. Journal of Cellular Biochemistry, 118(11), 4095–4108. https://doi.org/10.1002/jcb.26067 Osycka-Salut, C. E., Martínez-León, E., Gervasi, M. G., Castellano, L., Davio, C., Chiarante, N., Franchi, A. M., Ribeiro, M. L., Díaz, E. S., & Perez-Martinez, S. (2020). Fibronectin induces capacitation-associated events through the endocannabinoid system in bull sperm. Theriogenology, 153, 91–101. https://doi.org/10.1016/j.theriogenology.2020.04.031 Panzani, D., Rota, A., Tesi, M., Fanelli, D., & Camillo, F. (2018). Update on Donkey Embryo Transfer and Cryopreservation. Journal of Equine Veterinary Science, 65, 50–54. https://doi.org/10.1016/J.JEVS.2017.11.012 Patrat, C., Serres, C., & Jouannet, P. (2000). The acrosome reaction in human spermatozoa. Biology of the Cell, 92(3–4), 255–266. https://doi.org/10.1016/s0248-4900(00)01072-8 Peña-Alfaro, C. E., Barros, L. O., Carneiro, G. F., Gastal, M. O., & Gastal, E. L. (2014). Embryo transfer in Pega donkeys (Equus asinus) in Brazil. Journal of Equine Veterinary Science, 34(1), 185. https://doi.org/10.1016/J.JEVS.2013.10.135 Perez-Osorio, J., Mello, F. G., Juliani, G., Lagares, M., Lago, L., & Henry, M. (2008). Effect on post-thaw viability of equine sperm using stepwise addition of dimethyl formamide and varying cooling and freezing procedures. Animal Reproduction, 5(3–4), 103–109. Pizarro, E., Restrepo, G., Echeverry, J., & Rojano, B. (2013). Effect of seminal plasma on the redox state of cryopreserved stallion semen. Revista MVZ Córdoba, 18, 3672–3680. Quaresma, M., Martins, A. M. F., Rodrigues, J. B., Colaço, J., & Payan-Carreira, R. (2014). Pedigree and herd characterization of a donkey breed vulnerable to extinction. Animal, 8(3), 354–359. https://doi.org/10.1017/S1751731113002218 Rathi, R., Colenbrander, B., Bevers, M. M., & Gadella, B. M. (2001). Evaluation of in vitro capacitation of stallion spermatozoa. Biology of Reproduction, 65(2), 462–470. https://doi.org/10.1095/biolreprod65.2.462 Restrepo, G., Cantero Naclares, J. M., & Montoya Paez, J. D. (2016). Efecto De La Centrifugación Sobre La Integridad Y La Funcionalidad De Espermatozoides Equinos. Biotecnoloía En El Sector Agropecuario y Agroindustrial, 14(1), 119. https://doi.org/10.18684/bsaa(14)119-125 Restrepo, G., Pérez, Daniel., Acosta, Mariano., Camacho, C., & Pérez, J. (2017). Freezing of Equine Semen Under Two Schemes of Addition of Dimethylformamide. Revista de Investigaciones Veterinarias Del Peru, 28(4), 918–927. https://doi.org/10.15381/rivep.v28i4.13884 Restrepo, G., Zapata Acosta, K., & Rojano, B. (2015). Evaluación de la capacidad antioxidante total del plasma seminal equino. Zootecnia Tropical, 33(1), 79–87. Ribeiro-Peres, A., Munita-Barbosa, L., Yumi-Kanazawa, M., Mello-Martins, M. I., & Ferreira De Souza, F. (2014). Criopreservación de espermatozoides bovinos extraídos de la cola del epidídimo utilizando los métodos convencional y automatizado. Archivos de Medicina Veterinaria, 46(1), 31–38. https://doi.org/10.4067/S0301-732X2014000100005 Rodriguez, P. C., Satorre, M. M., & Beconi, M. T. (2012). Effect of two intracellular calcium modulators on sperm motility and heparin-induced capacitation in cryopreserved bovine spermatozoa. Animal Reproduction Science, 131(3–4), 135–142. https://doi.org/10.1016/j.anireprosci.2012.03.015 Rodríguez-Villamil, P., Mentz, D., Ongaratto, F. L., Aguiar, L. H., Rodrigues, J. L., Bertolini, M., & Moura, A. A. (2020). Assessment of binder of sperm protein 1 (BSP1) and heparin effects on in vitro capacitation and fertilization of bovine ejaculated and epididymal sperm. Zygote, 28(6), 489–494. https://doi.org/10.1017/S0967199420000374 Rossel, S., Marshall, F., Peters, J., Pilgram, T., Adams, M. D., & O’Connor, D. (2008). Domestication of the donkey: Timing, processes, and indicators. Proceedings of the National Academy of Sciences of the United States of America, 105(10), 3715–3720. https://doi.org/10.1073/pnas.0709692105 Rota, A., Sabatini, C., Przybył, A., Ciaramelli, A., Panzani, D., & Camillo, F. (2019). Post-thaw Addition of Caffeine and/or Pentoxifylline Affect Differently Motility of Horse and Donkey-Cryopreserved Spermatozoa. Journal of Equine Veterinary Science, 75, 41–47. https://doi.org/10.1016/j.jevs.2019.01.003 Sabatini, C., Mari, G., Mislei, B., Love, C. C., Panzani, D., Camillo, F., & Rota, A. (2014). Effect of Post-Thaw Addition of Seminal Plasma on Motility, Viability and Chromatin Integrity of Cryopreserved Donkey Jack (Equus asinus) Spermatozoa. Reproduction in Domestic Animals, 49(6), 989–994. https://doi.org/https://doi.org/10.1111/rda.12419 Sati, L., Cayli, S., Delpiano, E., Sakkas, D., & Huszar, G. (2014). The pattern of tyrosine phosphorylation in human sperm in response to binding to zona pellucida or hyaluronic acid. Reproductive Sciences, 21(5), 573–581. https://doi.org/10.1177/1933719113504467 Shahsavari, M. H., Moghaddam, G., Kia, H. D., & Rodrigues, A. P. R. (2019). Effects of new synthetic cryoprotectant agents on histological characteristics of various classes of vitrified bovine pre-antral follicles. Veterinary Research Forum, 10(1), 9–16. https://doi.org/10.30466/vrf.2019.34306 Shen, L., Chen, H., Zhu, Q., Wang, Y., Wang, S., Qian, J., Wang, Y., & Qu, H. (2016). Identification of bioactive ingredients with immuno-enhancement and anti-oxidative effects from Fufang-Ejiao-Syrup by LC-MSn combined with bioassays. Journal of Pharmaceutical and Biomedical Analysis, 117, 363–371. https://doi.org/10.1016/j.jpba.2015.09.024 Sinha, B., Semmler, J., Eisenhut, T., Eigler, A., & Endres, S. (1995). Enhanced tumor necrosis factor suppression and cyclic adenosine monophosphate accumulation by combination of phosphodiesterase inhibitors and prostanoids. European Journal of Immunology, 25(1), 147–153. https://doi.org/https://doi.org/10.1002/eji.1830250125 Sjunnesson, Y. (2020). In vitro fertilisation in domestic mammals—a brief overview. Upsala Journal of Medical Sciences, 125(2), 68–76. https://doi.org/10.1080/03009734.2019.1697911 Slanina, T., Miškeje, M., Tirpák, F., Błaszczyk, M., Formicki, G., & Massányi, P. (2018). Caffeine strongly improves motility parameters of Turkey spermatozoa with no effect on cell viability. Acta Veterinaria Hungarica, 66(1), 137–150. https://doi.org/10.1556/004.2018.013 Smits, K., Hoogewijs, M., Woelders, H., Daels, P., & Van Soom, A. (2012). Breeding or assisted reproduction? Relevance of the horse model applied to the conservation of endangered equids. Reproduction in Domestic Animals, 47(SUPPL.4), 239–248. https://doi.org/10.1111/j.1439-0531.2012.02082.x Soni, Y., Talluri, T. R., Kumar, A., Ravi, S. K., Mehta, J. S., & Tripathi, B. N. (2019). Effects of different concentration and combinations of cryoprotectants on sperm quality, functional integrity in three Indian horse breeds. Cryobiology, 86(August 2018), 52–57. https://doi.org/10.1016/j.cryobiol.2018.12.005 Stanic, P., Sonicki, Z., & Suchanek, E. (2002). Effect of pentoxifylline on motility and membrane integrity of cryopreserved human spermatozoa. International Journal of Andrology, 25(3), 186–190. https://doi.org/10.1046/j.1365-2605.2002.00348.x Stephens, T. D., Brooks, R. M., Carrington, J. L., Cheng, L., Carrington, A. C., Porr, C. A., & Splan, R. K. (2013). Effects of pentoxifylline, caffeine, and taurine on post-thaw motility and longevity of equine frozen semen. Journal of Equine Veterinary Science, 33(8), 615–621. https://doi.org/10.1016/j.jevs.2012.10.004 Strohmeyer, N., Bharadwaj, M., Costell, M., Fässler, R., & Müller, D. J. (2017). Fibronectin-bound α5β1 integrins sense load and signal to reinforce adhesion in less than a second. Nature Materials, 16(12), 1262–1270. https://doi.org/10.1038/nmat5023 Talukdar, D., Ahmed, K., Deori, S., & Das, G. C. (2015). Heparin-induced in vitro capacitation changes of swamp buffalo spermatozoa. Turkish Journal of Veterinary and Animal Sciences, 39(5), 629–633. https://doi.org/10.3906/vet-1501-16 Tash, J. S., & Means, A. R. (1982). Regulation of protein phosphorylation and motility of sperm by cyclic adenosine monophosphate and calcium. Biology of Reproduction, 26(4), 745–763. https://doi.org/10.1095/biolreprod26.4.745 The Donkey Sanctuary. (2017). Bajo la Piel: El comercio emergente de las pieles de burro y sus consecuencias para el bienestar y la subsistencia de estos animales. Thomas, A. D., Meyers, S. A., & Ball, B. A. (2006). Capacitation-like changes in equine spermatozoa following cryopreservation. Theriogenology, 65(8), 1531–1550. https://doi.org/10.1016/j.theriogenology.2005.08.022 Turley, E. A., Noble, P. W., & Bourguignon, L. Y. W. (2002). Signaling properties of hyaluronan receptors. In Journal of Biological Chemistry (Vol. 277, Issue 7, pp. 4589–4592). https://doi.org/10.1074/jbc.R100038200 Úsuga, A., Rojano, B. A., & Restrepo, G. (2018). Efecto de los componentes del plasma seminal en la calidad del semen equino. Revista CES Medicina Veterinaria y Zootecnia, 13, 281. Varela, E., Rojas, M., & Restrepo, G. (2020). Membrane stability and mitochondrial activity of bovine sperm frozen with low-density lipoproteins and trehalose. Reproduction in Domestic Animals = Zuchthygiene, 55(2), 146–153. https://doi.org/10.1111/rda.13599 Verma, A. S., Agrahari, S., Rastogi, S., & Singh, A. (2011). Biotechnology in the realm of history. In Journal of Pharmacy and Bioallied Sciences (Vol. 3, Issue 3, pp. 321–323). Wolters Kluwer -- Medknow Publications. https://doi.org/10.4103/0975-7406.84430 Vidament, M., Vincent, P., Martin, F. X., Magistrini, M., & Blesbois, E. (2009). Differences in ability of jennies and mares to conceive with cooled and frozen semen containing glycerol or not. Animal Reproduction Science, 112(1–2), 22–35. https://doi.org/10.1016/j.anireprosci.2008.03.016 Visconti, P. E. (2009). Understanding the molecular basis of sperm capacitation through kinase design. In Proceedings of the National Academy of Sciences of the United States of America (Vol. 106, Issue 3, pp. 667–668). National Academy of Sciences. https://doi.org/10.1073/pnas.0811895106 Warren, L. K. (2009). Feeding the Stallion. Agri-Facts, January, 1–4. White, I. G. (1993). Lipids and calcium uptake of sperm in relation to cold shock and preservation: a review. Reproduction, Fertility, and Development, 5(6), 639–658. https://doi.org/10.1071/rd9930639 Yilmaz, O., Boztepe, S., & Ertugrul, M. (2012). Domesticated Donkeys – Part II: Types and Breeds. Canadian Journal of Animal Science, 2012(April). Yovich, J. L. (1993). Pentoxifylline: actions and applications in assisted reproduction. Human Reproduction, 8(11), 1786–1791. https://doi.org/10.1093/oxfordjournals.humrep.a137935 Zamzami, N., Susin, S. A., Marchetti, P., Hirsch, T., Gómez-Monterrey, I., Castedo, M., & Kroemer, G. (1996). Mitochondrial control of nuclear apoptosis. Journal of Experimental Medicine, 183(4), 1533–1544. https://doi.org/10.1084/jem.183.4.1533 Zhao, Q., Tao, C., Pan, J., Wei, Q., Zhu, Z., Wang, L., Liu, M., Huang, J., Yu, F., Chen, X., Zhang, L., & Li, J. (2021). Equine chorionic gonadotropin pretreatment 15 days before fixed-time artificial insemination improves the reproductive performance of replacement gilts. Animal, 15(12), 100406. https://doi.org/10.1016/J.ANIMAL.2021.100406 Zheng, C., Jian-tao, Z., Hai-feng, L., & Hong-bin, W. (2016). Laparoscopic Embryo Transfer in Pigs: Knowledge for Surgical Procedures. Journal of Northeast Agricultural University (English Edition), 23(2), 52–58. https://doi.org/10.1016/S1006-8104(16)30047-2 |
| dc.rights.coar.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
| dc.rights.license.spa.fl_str_mv |
Reconocimiento 4.0 Internacional |
| dc.rights.uri.spa.fl_str_mv |
http://creativecommons.org/licenses/by/4.0/ |
| dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
| rights_invalid_str_mv |
Reconocimiento 4.0 Internacional http://creativecommons.org/licenses/by/4.0/ http://purl.org/coar/access_right/c_abf2 |
| eu_rights_str_mv |
openAccess |
| dc.format.extent.spa.fl_str_mv |
xi, 65 páginas |
| dc.format.mimetype.spa.fl_str_mv |
application/pdf |
| dc.publisher.spa.fl_str_mv |
Universidad Nacional de Colombia |
| dc.publisher.program.spa.fl_str_mv |
Medellín - Ciencias - Maestría en Ciencias - Biotecnología |
| dc.publisher.faculty.spa.fl_str_mv |
Facultad de Ciencias |
| dc.publisher.place.spa.fl_str_mv |
Medellín, Colombia |
| dc.publisher.branch.spa.fl_str_mv |
Universidad Nacional de Colombia - Sede Medellín |
| institution |
Universidad Nacional de Colombia |
| bitstream.url.fl_str_mv |
https://repositorio.unal.edu.co/bitstream/unal/83291/1/license.txt https://repositorio.unal.edu.co/bitstream/unal/83291/2/Tesis%20Burros%20Luis%20Mendoza%20Arenas.pdf https://repositorio.unal.edu.co/bitstream/unal/83291/3/Tesis%20Burros%20Luis%20Mendoza%20Arenas.pdf.jpg |
| bitstream.checksum.fl_str_mv |
eb34b1cf90b7e1103fc9dfd26be24b4a 0d23318ae0d8223fa1ee53914faa5139 e15f79b65d0c97fc6ed0350e009a6f2c |
| bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 |
| repository.name.fl_str_mv |
Repositorio Institucional Universidad Nacional de Colombia |
| repository.mail.fl_str_mv |
repositorio_nal@unal.edu.co |
| _version_ |
1814089451998019584 |
| spelling |
Reconocimiento 4.0 Internacionalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Restrepo Betancur, Giovannibd03434c72051e913ab3e03d3c37d88dMendoza Arenas, Luis Mendoza650cf780f6d4c72344ad77f045f3f634GIBA (Grupo de Investigación Biotecnología Animal)2023-02-03T18:35:52Z2023-02-03T18:35:52Z2022-02-02https://repositorio.unal.edu.co/handle/unal/83291Universidad Nacional de ColombiaRepositorio Institucional Universidad Nacional de Colombiahttps://repositorio.unal.edu.co/Las biotecnologías reproductivas, constituyen una serie de herramientas para la conservación de las razas consideradas en peligro de extinción como los burros. El objetivo de esta investigación fue evaluar el uso de inductores de capacitación espermática en semen congelado-descongelado de asnos. Se colectaron 5 asnos, localizados en el Valle de Aburrá con edades entre 4-8 años, se obtuvieron 3 eyaculados por animal, mediante vagina artificial, el semen recién recolectado fue diluido en proporción 1:1 en Equiplus® para ser criopreservado. Después, se descongelaron dos pajillas de 0.5 mL cada una del mismo eyaculado en un baño María a 37 °C, este volumen fue divido en 8 submuetras y 125 μL de semen fueron depositados en tubos eppendorf, posteriormente se adicionó a cada tubo el inductor de capacitación pentoxifilina (PTX) 2.5, 5 y 7.5 mM, ácido etilendiaminotetraacético (EDTA) 0.5, 1 y 1.5 mM, heparina (HEP) 1.1 mM como control positivo y sin aditivo (control) por 20 minutos. Se evaluaron variables de cinética espermática sistema IVOS (Hamilton Thorne), vitalidad espermática (VE) mediante el kit de viabilidad LIVE/DEAD, integridad acrosómica con (FITC-PSA), mientras que la capacitación, potencial mitocondrial e ingreso de Ca2+, fue evaluado usando un citómetro de flujo (FORTESSA LSR, BD Biosciences, EE. UU). Se observó que la movilidad total (MT) y la vitalidad espermática (VE) aumentaron en relación con el control, con el uso de 2,5 y 5.0 mM de PTX respectivamente (p<0.05), mientras que la movilidad progresiva (MP) disminuyó para las 3 concentraciones de EDTA (p<0.05), otras variables como velocidad lineal (VSL), velocidad curvilínea (VCL), amplitud lateral de la cabeza (ALH) y frecuencia de batida (BCF) disminuyen su proporción (p<0.05) al adicionar de EDTA 1, 1.5 y PTX 7.5 mM. En cuanto a la capacitación, se encontró que las concentraciones de EDTA 1, 1.5 mM y PTX 5, 7.5 mM (p<0.01) favorecen eventos asociados a la capacitación espermática y además disminuye la concentración de Ca2+ intracelular para todas las concentraciones de PTX y EDTA 0.5 mM (p<0.01). No se observaron efectos en la actividad mitocondrial. Se concluye que la adición de PTX a 2.5, 5 mM, incrementa la proporción de la MT y VE, por el contrario, la MP disminuye al aumentar la concentración de EDTA, además cambios en la estructura de la membrana plasmática asociados a eventos de capacitación de semen congelado-descongelado de burro, puede ser modulada por la suplementación con PTX (5 mM y 7.5 mM) y EDTA (1 mM y 1.5 mM). Sin embargo, la adición de estos inductores de la capacitación, no logró influencia en el ingreso de Ca2+, por lo tanto, se requiere mayor nivel de investigación ya que los mecanismos e implicaciones en la capacitación espermática para semen de burro aún son desconocidos. (Tomado de la fuente)Reproductive biotechnologies constitute a series of tools for the conservation of breeds considered in danger of extinction, such as donkeys. The objective of this research was to evaluate the use of sperm capacitation inducers in frozen-thawed donkey semen. Five donkeys were collected, located in the Valle de Aburrá with ages between 4-8 years, 3 ejaculates were obtained per animal, by means of an artificial vagina, the recently collected semen was diluted in a 1:1 ratio in Eplus® to be cryopreserved. Afterwards, two straws of 0.5 mL each of the same ejaculate were thawed in a water bath at 37 °C, this volume was divided into 8 sub-samples and 125 μL of semen were deposited in eppendorf tubes, later the sperm inducer was added to each tube. capacitation pentoxifylline (PTX) 2.5, 5 and 7.5 mM, ethylenediaminetetraacetic acid (EDTA) 0.5, 1 and 1.5 mM, heparin (HEP) 1.1 mM as positive control and without additive (control) for 20 minutes. Sperm kinetic variables IVOS system (Hamilton Thorne), sperm vitality (SV) using the LIVE/DEAD viability kit, acrosomal integrity with (FITC-PSA), while capacitation, mitochondrial potential and Ca2+ ingress were evaluated. using a flow cytometer (FORTESSA LSR, BD Biosciences, USA). It was observed that the total motility (MT) and the sperm vitality (VE) increased in relation to the control, with the use of 2.5 and 5.0 mM of PTX respectively (p<0.05), while the progressive motility (MP) decreased for the 3 concentrations of EDTA (p<0.05), other variables such as linear velocity (VSL), curvilinear velocity (VCL), lateral amplitude of the head (ALH) and beat frequency (BCF) decrease its proportion (p<0.05) by adding EDTA 1, 1.5 and PTX 7.5 mM. Regarding capacitation, it was found that concentrations of EDTA 1, 1.5 mM and PTX 5, 7.5 mM (p<0.01) favor events associated with sperm capacitation and also decrease intracellular Ca2+ concentration for all concentrations of PTX and 0.5mM EDTA (p<0.01). No effects on mitochondrial activity were observed. It is concluded that the addition of PTX at 2.5, 5 mM, increases the proportion of MT and VE, on the contrary, MP decreases with increasing concentration of EDTA, in addition to changes in the structure of the plasma membrane associated with capacitation events. of frozen-thawed donkey semen, can be modulated by supplementation with PTX (5 mM and 7.5 mM) and EDTA (1 mM and 1.5 mM). However, the addition of these capacitation inducers did not influence Ca2+ entry, therefore, a higher level of research is required since the mechanisms and implications in sperm capacitation for donkey semen are still unknown.MaestríaMagíster en Ciencias - BiotecnologíaReproducción AnimalÁrea curricular Biotecnologíaxi, 65 páginasapplication/pdfspaUniversidad Nacional de ColombiaMedellín - Ciencias - Maestría en Ciencias - BiotecnologíaFacultad de CienciasMedellín, ColombiaUniversidad Nacional de Colombia - Sede Medellín590 - AnimalesConservación de semenAnimales - Conducta sexualPentoxifilinaBiotecnologíaBiotechnologyPentoxifyllineEvaluación de inductores de la capacitación espermática en semen criopreservado de asnos (equus asinus)Evaluation of spermatic capacity inducers in criopreserved semen of donkeys (Equus asinus)Trabajo de grado - Maestríainfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/acceptedVersionTexthttp://purl.org/redcol/resource_type/TMLaReferenciaAcha, D., Hidalgo, M., Ortiz, I., Gálvez, M. J., Carrasco, J. J., Gómez-Arrones, V., & Dorado, J. (2016). Freezability of Andalusian donkey (Equus asinus) spermatozoa: Effect of extenders and permeating cryoprotectants. Reproduction, Fertility and Development, 28(12), 1990–1998. https://doi.org/10.1071/RD14449Arenas, E., Cambron, A., Ambriz, D., Zuñiga, P., Rodriguez, A., & Rosado, A. (2010). Bases fisiologicas de la capacitacion y de la reaccion acrosomal del espermatozoide. Contactos, 78, 5–11.Ball, G. F., & Balthazart, J. (2008). How useful is the appetitive and consummatory distinction for our understanding of the neuroendocrine control of sexual behavior? In Hormones and Behavior (Vol. 53, Issue 2, pp. 307–311). NIH Public Access. https://doi.org/10.1016/j.yhbeh.2007.09.023Barakat, I. A. H., Danfour, M. A., Galewan, F. A. M., & Dkhil, M. A. (2015). Effect of Various Concentrations of Caffeine, Pentoxifylline, and Kallikrein on Hyperactivation of Frozen Bovine Semen. BioMed Research International, 2015, 948575. https://doi.org/10.1155/2015/948575Bennett, R., & Pfuderer, S. (2020). The potential for new donkey farming systems to supply the growing demand for hides. Animals, 10(4). https://doi.org/10.3390/ani10040718Bhattacharya, S. (2018). Cryopretectants and Their Usage in Cryopreservation Process. Cryopreservation Biotechnology in Biomedical and Biological Sciences. https://doi.org/10.5772/intechopen.80477Blench, R. (2012). Wild Asses and Donkeys in Africa: Interdisciplinary Evidence for Their Biogeography, History and Current Use. Soas, May 2012.Bollwein, H., Fuchs, I., & Koess, C. (2008). Interrelationship Between Plasma Membrane Integrity, Mitochondrial Membrane Potential and DNA Fragmentation in Cryopreserved Bovine Spermatozoa. Reproduction in Domestic Animals, 43(2), 189–195. https://doi.org/10.1111/j.1439-0531.2007.00876.xBoni, R., Gallo, A., & Cecchini, S. (2017). Kinetic activity, membrane mitochondrial potential, lipid peroxidation, intracellular pH and calcium of frozen/thawed bovine spermatozoa treated with metabolic enhancers. Andrology, 5(1), 133–145. https://doi.org/10.1111/andr.12259Breitbart, H., & Finkelstein, M. (2015). Regulation of Sperm Capacitation and the Acrosome Reaction by PIP2 and Actin Modulation. Asian Journal of Andrology, 17(4), 597–600. https://doi.org/10.4103/1008-682X.154305Breitbart, H., & Shabtay, O. (2018). Sperm acrosome reaction. In Encyclopedia of Reproduction (Second Edi, Vol. 3). Elsevier. https://doi.org/10.1016/B978-0-12-801238-3.66186-XBrener, E., Rubinstein, S., Cohen, G., Shternall, K., Rivlin, J., & Breitbart, H. (2003). Remodeling of the actin cytoskeleton during mammalian sperm capacitation and acrosome reaction. Biology of Reproduction, 68(3), 837–845. https://doi.org/10.1095/biolreprod.102.009233Buffone, M. G., Wertheimer, E. V., Visconti, P. E., & Krapf, D. (2014). Central role of soluble adenylyl cyclase and cAMP in sperm physiology. In Biochimica et Biophysica Acta - Molecular Basis of Disease (Vol. 1842, Issue 12, pp. 2610–2620). Elsevier. https://doi.org/10.1016/j.bbadis.2014.07.013Bustamante, I. C., Pederzolli, C. D., Sgaravatti, A. M., Gregory, R. M., Dutra Filho, C. S., Jobim, M. I. M., & Mattos, R. C. (2009). Skim milk-egg yolk based semen extender compensates for non-enzymatic antioxidant activity loss during equine semen cryopreservation. In Anim. Reprod., v (Vol. 6, Issue 2). Colégio Brasileiro de Reprodução Animal.Calogero, A. E., Fishel, S., Hall, J., Ferrara, E., Vicari, E., Green, S., Hunter, A., Burrello, N., Thornton, S., & D’Agata, R. (1998). Correlation between intracellular cAMP content, kinematic parameters and hyperactivation of human spermatozoa after incubation with pentoxifylline. Human Reproduction, 13(4), 911–915. https://doi.org/10.1093/humrep/13.4.911Canisso, I. F., Carvalho, G. R., Morel, M. C. G. D., Guimarães, J. D., & McDonnell, S. M. (2010). Sexual behavior and ejaculate characteristics in Pêga donkeys (Equus asinus) mounting estrous horse mares (Equus caballus). Theriogenology, 73(1), 56–63. https://doi.org/10.1016/j.theriogenology.2009.07.026Canisso, I. F., & McDonnell, S. M. (2010). Donkey breeding behavior with an emphasis on the Pêga breed. MATTHEWS NS &TAYLOR TS.Veterinary Care of Donkeys.Ithaca: International Veterinary Information Service, 310.Canisso, I. F., Panzani, D., Miró, J., & Ellerbrock, R. E. (2019). Key Aspects of Donkey and Mule Reproduction. Veterinary Clinics of North America - Equine Practice, 35(3), 607–642. https://doi.org/10.1016/j.cveq.2019.08.014Carluccio, A., Contri, A., Amendola, S., De Angelis, E., De Amicis, I., & Mazzatenta, A. (2013). Male isolation: A behavioral representation of the pheromonal “female effect” in donkey (Equus asinus). Physiology and Behavior, 118, 1–7. https://doi.org/10.1016/j.physbeh.2013.04.005Carminati, D., & Tidona, F. (2017). Nutritional Value and Potential Health Benefits of Donkey Milk. In Nutrients in Dairy and Their Implications for Health and Disease. Elsevier Inc. https://doi.org/10.1016/B978-0-12-809762-5.00031-0Carneiro, G. F., Cavalcante Lucena, J. E., & de Oliveira Barros, L. (2018). The Current Situation and Trend of the Donkey Industry in South America. Journal of Equine Veterinary Science, 65, 106–110. https://doi.org/10.1016/j.jevs.2018.03.007Carvalho, L. E., Silva Filho, J. M., Palhares, M. S., Sales, A. L. R., Gonczarowska, A. T., Oliveira, H. N., Resende, M. C., Neves, M. G., & Madison, R. J. (2017). Physical characteristics and fertility of fractionated donkey semen cooled at 5°C. Arquivo Brasileiro de Medicina Veterinaria e Zootecnia, 69(1), 29–38. https://doi.org/10.1590/1678-4162-7976Consuegra, C., Crespo, F., Bottrel, M., Ortiz, I., Dorado, J., Diaz-Jimenez, M., Pereira, B., & Hidalgo, M. (2018). Stallion sperm freezing with sucrose extenders: A strategy to avoid permeable cryoprotectants. Animal Reproduction Science, 191, 85–91. https://doi.org/10.1016/j.anireprosci.2018.02.013Contri, A., Gloria, A., Robbe, D., De Amicis, I., & Carluccio, A. (2012a). Characteristics of donkey spermatozoa along the length of the epididymis. Theriogenology, 77(1), 166–173. https://doi.org/10.1016/j.theriogenology.2011.07.031Contri, A., Gloria, A., Robbe, D., Sfirro, M. P., & Carluccio, A. (2012b). Effect of sperm concentration on characteristics of frozen-thawed semen in donkeys. Animal Reproduction Science, 136(1–2), 74–80. https://doi.org/10.1016/j.anireprosci.2012.10.022Cortés-Gutiérrez, E. I., Dávila-Rodríguez, M. I., López-Fernández, C., Fernández, J. L., Crespo, F., & Gosálvez, J. (2014). Localization of alkali-labile sites in donkey (Equus asinus) and stallion (Equus caballus) spermatozoa. Theriogenology, 81(2), 321–325. https://doi.org/10.1016/J.THERIOGENOLOGY.2013.10.001Darwin, C. (1859). Origem das espécies: E a seleção natural.Davoodi, F., Raisi, A., Rajabzadeh, A., Hablolvarid, M. H., & Zakian, A. (2020). The effects of verapamil and heparin co-administration on sperm parameters and oxidative stress in prevention of testicular torsion/detorsion damage in rats. Andrologia, 52(2), 1–8. https://doi.org/10.1111/and.13479de Andrade, A. F. C., Zaffalon, F. G., Celeghini, E. C. C., Nascimento, J., Tarragó, O. F. B., Martins, S., Alonso, M. A., & Arruda, R. P. (2011). Addition of Seminal Plasma to Post-thawing Equine Semen: What is the Effect on Sperm Cell Viability? Reproduction in Domestic Animals, 46(4), 682–686. https://doi.org/https://doi.org/10.1111/j.1439-0531.2010.01729.xde Vasconcelos Franco, J. S., Faheem, M., Chaveiro, A., & Moreira da Silva, F. (2016). Effects of α-tocopherol and freezing rates on the quality and heterologous in vitro fertilization capacity of stallion sperm after cryopreservation. Theriogenology, 86(4), 957–962. https://doi.org/10.1016/j.theriogenology.2016.03.019Demyda-Peyrás, S., Bottrel, M., Acha, D., Ortiz, I., Hidalgo, M., Carrasco, J. J., Gómez-Arrones, V., Gósalvez, J., & Dorado, J. (2018). Effect of cooling rate on sperm quality of cryopreserved Andalusian donkey spermatozoa. Animal Reproduction Science, 193, 201–208. https://doi.org/10.1016/j.anireprosci.2018.04.069Dhanju, C. K., Toor, G., & Cheema, R. S. (2009). Effect of seminal plasma on quantily and qualily of proteins leaked during heparin - induced in vitro capacitation of bull sperm. Indian Journal of Animal Research, 43(2), 136–138.Di Palma, T., Cecchini, S., MacChia, G., Pasolini, M. P., Cocchia, N., & Boni, R. (2020). Kinematic, bioenergetic and oxidative evaluations of donkey sperm preserved at +4°C. Zygote, 1–8. https://doi.org/10.1017/S096719942000012XDi Paolo, G., & De Camilli, P. (2006). Phosphoinositides in cell regulation and membrane dynamics. Nature, 443(7112), 651–657. https://doi.org/10.1038/nature05185Di Santo, M., Tarozzi, N., Nadalini, M., & Borini, A. (2012). Human sperm cryopreservation: Update on techniques, effect on DNA integrity, and implications for ART. Advances in Urology, 2012. https://doi.org/10.1155/2012/854837Diaz-Jimenez, M., Dorado, J., Ortiz, I., Consuegra, C., Pereira, B., Gonzalez-De Cara, C. A., Aguilera, R., Mari, G., Mislei, B., Love, C. C., & Hidalgo, M. (2018). Cryopreservation of donkey sperm using non-permeable cryoprotectants. Animal Reproduction Science, 189(December 2017), 103–109. https://doi.org/10.1016/j.anireprosci.2017.12.013Dong, W. W., Zeng, Z. Da, Au, D., Chan, C. O., Yang, D. J., & Li, X. B. (2012). Quality control of Colla corii asini using near-infrared spectroscopy and chemometrics clustering techniques. Journal of Food and Drug Analysis, 20(1), 152–158.Dorado, J., Acha, D., Ortiz, I., Gálvez, M. J., Carrasco, J. J., Díaz, B., Gómez-Arrones, V., Calero-Carretero, R., & Hidalgo, M. (2013). Relationship between conventional semen characteristics, sperm motility patterns and fertility of Andalusian donkeys (Equus asinus). Animal Reproduction Science, 143(1–4), 64–71. https://doi.org/10.1016/j.anireprosci.2013.10.003Dorado, J., Hidalgo, M., Acha, D., Ortiz, I., Bottrel, M., Azcona, F., Carrasco, J. J., Gómez-Arrones, V., & Demyda-Peyrás, S. (2019). Cryopreservation of Andalusian donkey (Equus asinus) spermatozoa: Use of alternative energy sources in the freezing extender affects post-thaw sperm motility patterns but not DNA stability. Animal Reproduction Science, 208(June), 106126. https://doi.org/10.1016/j.anireprosci.2019.106126Douet, C., Reigner, F., Barrière, P., Blard, T., Deleuze, S., & Goudet, G. (2019). First attempts for vitrification of immature oocytes in donkey (Equus asinus): Comparison of two vitrification methods. Theriogenology, 126, 261–265. https://doi.org/10.1016/j.theriogenology.2018.12.030El-Shahat, K. H., Taysser, M. I., Badr, M. R., & Zaki, K. A. (2016). Effect of heparin, caffeine and calcium ionophore A23187 on in vitro induction of the acrosome reaction of fresh ram spermatozoa. Asian Pacific Journal of Reproduction, 5(2), 148–155. https://doi.org/10.1016/j.apjr.2016.01.012Esteves, S. C., Spaine, D. M., & Cedenho, A. P. (2007). Effects of pentoxifylline treatment before freezing on motility, viability and acrosome status of poor quality human spermatozoa cryopreserved by the liquid nitrogen vapor method. Brazilian Journal of Medical and Biological Research, 40(7), 985–992. https://doi.org/10.1590/S0100-879X2006005000118Fernández, S., Morado, S., Cetica, P., & Córdoba, M. (2020). Hyaluronic acid capacitation induces intracellular signals modulated by membrane-associated adenylate cyclase and tyrosine kinase involved in bovine in vitro fertilization. Theriogenology, 148, 174–179. https://doi.org/10.1016/j.theriogenology.2020.02.033Ferrante, A., Baca Castex, C., Bruno, S., Arraztoa, C., Plaza, J., Neild, D., & Miragaya, M. (2018a). Comparison of Whole and Centrifuged Egg Yolk Added to Kenney’s and Lactose-EDTA Extenders for Donkey Semen Cryopreservation. Journal of Equine Veterinary Science, 65, 12–18. https://doi.org/10.1016/j.jevs.2018.02.024Ferrante, A., Baca Castex, C., Bruno, S., Arraztoa, C., Plaza, J., Neild, D., & Miragaya, M. (2018b). Comparison of Whole and Centrifuged Egg Yolk Added to Kenney’s and Lactose-EDTA Extenders for Donkey Semen Cryopreservation. Journal of Equine Veterinary Science, 65, 12–18. https://doi.org/10.1016/J.JEVS.2018.02.024Ferré, L. B., Kjelland, M. E., Strøbech, L. B., Hyttel, P., Mermillod, P., & Ross, P. J. (2020). Review: Recent advances in bovine in vitro embryo production: reproductive biotechnology history and methods. Animal, 14(5), 991–1004. https://doi.org/10.1017/S1751731119002775Finkelstein, M., Etkovitz, N., & Breitbart, H. (2010). Role and regulation of sperm gelsolin prior to fertilization. The Journal of Biological Chemistry, 285(51), 39702–39709. https://doi.org/10.1074/jbc.M110.170951Finkelstein, M., Megnagi, B., Ickowicz, D., & Breitbart, H. (2013). Regulation of sperm motility by PIP2(4,5) and actin polymerization. Developmental Biology, 381(1), 62–72. https://doi.org/10.1016/j.ydbio.2013.06.014Fredholm, B. B., & Lindström, K. (1986). The Xanthine Derivative 1-(5′-Oxohexyl)-3-methyl-7-propyl Xanthine (HWA 285) Enhances the Actions of Adenosine. Acta Pharmacologica et Toxicologica, 58(3), 187–192. https://doi.org/https://doi.org/10.1111/j.1600-0773.1986.tb00093.xGamboa, S., Rodrigues, A. S., Henriques, L., Batista, C., & Ramalho-Santos, J. (2010). Seasonal functional relevance of sperm characteristics in equine spermatozoa. Theriogenology, 73(7), 950–958. https://doi.org/10.1016/j.theriogenology.2009.11.023García-Álvarez, O., Maroto-Morales, A., Jiménez-Rabadán, P., Ramón, M., del Olmo, E., Iniesta-Cuerda, M., Anel-López, L., Fernández-Santos, M. R., Garde, J. J., & Soler, A. J. (2015). Effect of different media additives on capacitation of frozen–thawed ram spermatozoa as a potential replacement for estrous sheep serum. Theriogenology, 84(6), 948–955. https://doi.org/10.1016/J.THERIOGENOLOGY.2015.05.032Gervasi, M. G., Marczylo, T. H., Lam, P. M., Rana, S., Franchi, A. M., Konje, J. C., & Perez-Martinez, S. (2013). Anandamide Levels Fluctuate in the Bovine Oviduct during the Oestrous Cycle. PLoS ONE, 8(8), e72521. https://doi.org/10.1371/journal.pone.0072521Gervasi, M. G., & Visconti, P. E. (2016). Chang’s meaning of capacitation: A molecular perspective. Molecular Reproduction and Development, 83(10), 860–874. https://doi.org/10.1002/mrd.22663Ghasemzadeh, A., Karkon-Shayan, F., Yousefzadeh, S., Naghavi-Behzad, M., & Hamdi, K. (2016). Study of pentoxifylline effects on motility and viability of spermatozoa from infertile asthenozoospermic males. Nigerian Medical Journal, 57(6), 324. https://doi.org/10.4103/0300-1652.193857Glogowski, J., Danforth, D. R., & Ciereszko, A. (2002). Inhibition of alkaline phosphatase activity of boar semen by pentoxifylline, caffeine, and theophylline. Journal of Andrology, 23(6), 783–792.Gloria, A., Contri, A., De Amicis, I., Robbe, D., & Carluccio, A. (2011). Differences between epididymal and ejaculated sperm characteristics in donkey. Animal Reproduction Science, 128(1–4), 117–122. https://doi.org/10.1016/j.anireprosci.2011.09.007Grötter, L. G., Cattaneo, L., Marini, P. E., Kjelland, M. E., & Ferré, L. B. (2019). Recent advances in bovine sperm cryopreservation techniques with a focus on sperm post-thaw quality optimization. Reproduction in Domestic Animals, 54(4), 655–665. https://doi.org/10.1111/rda.13409Guasti, P. N., Monteiro, G. A., Maziero, R. R. D., Carmo, M. T., Dell’Aqua, J. A., Crespilho, A. M., Rifai, E. A., & Papa, F. O. (2017). Pentoxifylline effects on capacitation and fertility of stallion epididymal sperm. Animal Reproduction Science, 179, 27–34. https://doi.org/10.1016/j.anireprosci.2017.01.013Guo, S., Savolainen, P., Su, J., Zhang, Q., Qi, D., Zhou, J., Zhong, Y., Zhao, X., & Liu, J. (2006). Origin of mitochondrial DNA diversity of domestic yaks. BMC Evolutionary Biology, 6, 73. https://doi.org/10.1186/1471-2148-6-73Haddad, R., Khan, R., Takahashi, Y. K., Mori, K., Harel, D., & Sobel, N. (2008). A metric for odorant comparison. Nature Methods, 5(5), 425–429. https://doi.org/10.1038/nmeth.1197Henry, M., Snoeck, P. P. N., & Cottorello, A. C. P. (2002). Post-thaw spermatozoa plasma membrane integrity and motility of stallion semen frozen with different cryoprotectants. Theriogenology, 58(2–4), 245–248. https://doi.org/10.1016/S0093-691X(02)00750-1Hu, J., Zhong, C., Ding, C., Chi, Q., Walz, A., Mombaerts, P., Matsunami, H., & Luo, M. (2007). Detection of near-atmospheric concentrations of CO2 by an olfactory subsystem in the mouse. Science, 317(5840), 953–957. https://doi.org/10.1126/science.1144233Hussain, N., Andrabi, S. M. H., & Mehmood, M. U. (2019). Effect of EDTA as Chelating Agent in Extender on the Post Thaw Quality of Buffalo Bull Spermatozoa. Cryo Letters, 40(3), 159–163.Itach, S. B.-S., Finklestein, M., Etkovitz, N., & Breitbart, H. (2012). Hyper-activated motility in sperm capacitation is mediated by phospholipase D-dependent actin polymerization. Developmental Biology, 362(2), 154–161. https://doi.org/10.1016/j.ydbio.2011.12.002Kaneko, T., & Nakagata, N. (2006). Improvement in the long-term stability of freeze-dried mouse spermatozoa by adding of a chelating agent. Cryobiology, 53(2), 279–282. https://doi.org/10.1016/J.CRYOBIOL.2006.06.004Keshtgar, S., Iravanpour, F., Gharesi-Fard, B., & Kazerooni, M. (2016). Combined effect of trolox and EDTA on frozen-thawed sperm quality. Iranian Journal of Medical Sciences, 41(3), 230–237.Keverne, E. B. (2002). Pheromones, vomeronasal function, and gender-specific behavior. In Cell (Vol. 108, Issue 6, pp. 735–738). Cell Press. https://doi.org/10.1016/S0092-8674(02)00687-6Kugler, W., Grunenfelder, H. P., & Broxham, E. (2008). Donkey Breeds in Europe: Inventory, Description, Need for Action, Conservation. In … and Seeds in Europe.Kumeta, Y., Maruyama, T., Asama, H., Yamamoto, Y., Hakamatsuka, T., & Goda, Y. (2014). Species identification of Asini Corii Collas (donkey glue) by PCR amplification of cytochrome b gene. Journal of Natural Medicines, 68(1), 181–185. https://doi.org/10.1007/s11418-013-0790-zLeclerc, P., Sirard, M. A., Chafouleas, J. G., & Lambert, R. D. (1992). Decrease in calmodulin concentrations during heparin-induced capacitation in bovine spermatozoa. Journal of Reproduction and Fertility, 94(1), 23–32. https://doi.org/10.1530/jrf.0.0940023Li, N., Yu, J., Yang, F., Shao, Y., Wu, S., Liu, B., Li, M., Wang, T., Li, J., & Zeng, S. (2021). L-Proline: An Effective Agent for Frozen and Post-thawed Donkey Semen Storage. Journal of Equine Veterinary Science, 101, 103393. https://doi.org/10.1016/j.jevs.2021.103393López, L. F. M. (2018). El puente de Occidente y la inegración de Antioquia. Universidad Nacional de Colombia-Sede Medellín.Mackenna, A. (1995). Contribution of the male factor to unexplained infertility: a review. International Journal of Andrology, 18 Suppl 1, 58–61. https://doi.org/10.1111/j.1365-2605.1995.tb00640.xMarshall, F. (2007). African pastoral perspectives on domestication of the donkey. In In Rethinking agriculture: archaeological and ethnoarchaeological perspectives (eds T. P. Denham, J. Iriarte & L. Vrydaghs) (Walnut Cre, pp. 371–407).Martínez-León, E., Osycka-Salut, C., Signorelli, J., Pozo, P., Pérez, B., Kong, M., Morales, P., Pérez-Martínez, S., & Díaz, E. S. (2015). Fibronectin stimulates human sperm capacitation through the cyclic AMP/protein kinase A pathway. Human Reproduction, 30(9), 2138–2151. https://doi.org/10.1093/humrep/dev154Massip, A. (2001). Cryopreservation of embryos of farm animals. In Reproduction in Domestic Animals (Vol. 36, Issue 2, pp. 49–55). John Wiley & Sons, Ltd. https://doi.org/10.1046/j.1439-0531.2001.00248.xMazur, P., Leibo, S. P., & Chu, E. H. Y. (1972). A two-factor hypothesis of freezing injury. Evidence from Chinese hamster tissue-culture cells. Experimental Cell Research, 71(2), 345–355. https://doi.org/10.1016/0014-4827(72)90303-5McDonnell, S. (2003). A Practical Field Guide to Horse Behavior: The Equid Ethogram. Journal of Equine Veterinary Science, 23(1), A1. https://doi.org/10.1016/s0737-0806(03)70087-2McLean, A. K., Navas González, F. J., & Canisso, I. F. (2019). Donkey and Mule Behavior. Veterinary Clinics of North America - Equine Practice, 35(3), 575–588. https://doi.org/10.1016/j.cveq.2019.08.010Medeiros, A. S. L., Gomes, G. M., Carmo, M. T., Papa, F. O., & Alvarenga, M. A. (2002). Cryopreservation of stallion sperm using different amides. Theriogenology, 58(2–4), 273–276. https://doi.org/10.1016/S0093-691X(02)00898-1Mendoza, C., Carreras, 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. Journal of Reproduction and Fertility, 95(3), 755–763. https://doi.org/10.1530/jrf.0.0950755Mercati, F., Domingo, P., Pasquariello, R., Dall’Aglio, C., Di Michele, A., Forti, K., Cocci, P., Boiti, C., Gil, L., Zerani, M., & Maranesi, M. (2020). Effect of chelating and antioxidant agents on morphology and DNA methylation in freeze-drying rabbit (Oryctolagus cuniculus) spermatozoa. Reproduction in Domestic Animals, 55(1), 29–37. https://doi.org/10.1111/rda.13577Mesa, A. M., Sc, M., R, G. H., & Sc, M. (2012). Efecto del colesterol y la dimetilformamida sobre parámetros posdescongelación en espermatozoides de caballos criollos colombianos Effect of cholesterol and dimethiyl-formamide on post-thawing parameters in Colombian creole stallion sperm. 17(1), 2908–2915.Meskini, N., Némoz, G., Okyayuz-Baklouti, I., Lagarde, M., & Prigent, A. F. (1994). Phosphodiesterase inhibitory profile of some related xanthine derivatives pharmacologically active on the peripheral microcirculation. Biochemical Pharmacology, 47(5), 781–788. https://doi.org/10.1016/0006-2952(94)90477-4Miragaya, M. H., Chaves, M. G., & Agüero, A. (2006). Reproductive biotechnology in South American camelids. Small Ruminant Research, 61(2–3), 299–310. https://doi.org/10.1016/J.SMALLRUMRES.2005.07.017Miragaya, M. H., Neild, D. M., & Alonso, A. E. (2018). A Review of Reproductive Biology and Biotechnologies in Donkeys. Journal of Equine Veterinary Science, 65, 55–61. https://doi.org/10.1016/j.jevs.2017.12.005Miró, J., Taberner, E., Rivera, M., Peña, A., Medrano, A., Rigau, T., & 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(8), 1017–1022. https://doi.org/10.1016/j.theriogenology.2009.06.012Mirshokraei, P., Hassanpour, H., Mehdizadeh, A., & Akhavan Taheri, M. (2011). Pentoxifylline induces capacitation and acrosome reaction and improves quality of motility in canine ejaculated spermatozoa. Research in Veterinary Science, 91(2), 281–284. https://doi.org/10.1016/J.RVSC.2011.01.002Molina, L. C. P., Luque, G. M., Balestrini, P. A., Marín-Briggiler, C. I., Romarowski, A., & Buffone, M. G. (2018). Molecular basis of human sperm capacitation. Frontiers in Cell and Developmental Biology, 6(July), 1–23. https://doi.org/10.3389/fcell.2018.00072Montoya, J. D., Rojano, B. A., & Betancur, G. R. (2017). Efecto de la suplementación del diluyente sobre la calidad del semen de asno a la descongelación. Archivos de Zootecnia.Nannou, T. K., Jouhara, H., Trembley, J., & Herrmann, J. (2016). Cryopreservation: Methods, equipment and critical concerns. Refrigeration Science and Technology, 22-25-June, 247–258. https://doi.org/10.18462/IIR.ICCRT.2016.0020Nassar, A., Mahony, M., Blackmore, P., Morshedi, M., Ozgur, K., & Oehninger, S. (1998). Increase of intracellular calcium is not a cause of pentoxifylline- induced hyperactivated motility or acrosome reaction in human sperm. Fertility and Sterility, 69(4), 748–754. https://doi.org/10.1016/S0015-0282(98)00013-2Naz, R. K., & Rajesh, P. B. (2004). Role of tyrosine phosphorylation in sperm capacitation/acrosome reaction. Reproductive Biology and Endocrinology, 2, 1–12. https://doi.org/10.1186/1477-7827-2-75Negri, P., Grechi, E., Tomasi, A., Fabbri, E., & Capuzzo, A. (1996). Effectiveness of pentoxifylline in semen preparation for intrauterine insemination. Human Reproduction, 11(6), 1236–1239. https://doi.org/10.1093/oxfordjournals.humrep.a019363Nguyen, V. K., Somfai, T., Salamone, D., Thu Huong, V. T., Le Thi Nguyen, H., Huu, Q. X., Hoang, A. T., Phan, H. T., Thi Pham, Y. K., & Pham, L. D. (2021). Optimization of donor cell cycle synchrony, maturation media and embryo culture system for somatic cell nuclear transfer in the critically endangered Vietnamese Ỉ pig. Theriogenology, 166, 21–28. https://doi.org/10.1016/J.THERIOGENOLOGY.2021.02.008Nouri, H., Towhidi, A., Zhandi, M., & Sadeghi, R. (2013). The Effects of Centrifuged Egg Yolk Used with INRA Plus Soybean Lecithin Extender on Semen Quality to Freeze Miniature Caspian Horse Semen. Journal of Equine Veterinary Science, 33(12), 1050–1053. https://doi.org/https://doi.org/10.1016/j.jevs.2013.03.184O’Connell, M., McClure, N., & Lewis, S. E. M. (2002). The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Human Reproduction, 17(3), 704–709. https://doi.org/10.1093/humrep/17.3.704Okazaki, T., Yoshida, S., Teshima, H., & Shimada, M. (2011). The addition of calcium ion chelator, EGTA to thawing solution improves fertilizing ability in frozen-thawed boar sperm. Animal Science Journal = Nihon Chikusan Gakkaiho, 82(3), 412–419. https://doi.org/10.1111/j.1740-0929.2010.00856.xOliveira, J. V. de, Oliveira, P. V. de L. F., Melo e Oña, C. M., Guasti, P. N., Monteiro, G. A., Sancler da Silva, Y. F. R., Papa, P. de M., Alvarenga, M. A., Dell’Aqua Junior, J. A., & Papa, F. O. (2016). Strategies to improve the fertility of fresh and frozen donkey semen. Theriogenology, 85(7), 1267–1273. https://doi.org/10.1016/j.theriogenology.2015.12.010Orrego, M. T., Melian, S. I., Montenegro, J., Cimato, A. N., Cisale, H., & Piehl, L. L. (2019). Boar sperm protein tyrosine phosphorylation in the presence of egg yolk soluble and low density lipoprotein fractions during cooling. Theriogenology, 123, 151–158. https://doi.org/10.1016/j.theriogenology.2018.09.031Ortgies, F., Klewitz, J., Görgens, A., Martinsson, G., & Sieme, H. (2012). Effect of procaine, pentoxifylline and trolox on capacitation and hyperactivation of stallion spermatozoa. Andrologia, 44(SUPPL.1), 130–138. https://doi.org/10.1111/j.1439-0272.2010.01150.xOsycka-Salut, C. E., Castellano, L., Fornes, D., Beltrame, J. S., Alonso, C. A. I., Jawerbaum, A., Franchi, A., Díaz, E. S., & Perez Martinez, S. (2017). Fibronectin From Oviductal Cells Fluctuates During the Estrous Cycle and Contributes to Sperm–Oviduct Interaction in Cattle. Journal of Cellular Biochemistry, 118(11), 4095–4108. https://doi.org/10.1002/jcb.26067Osycka-Salut, C. E., Martínez-León, E., Gervasi, M. G., Castellano, L., Davio, C., Chiarante, N., Franchi, A. M., Ribeiro, M. L., Díaz, E. S., & Perez-Martinez, S. (2020). Fibronectin induces capacitation-associated events through the endocannabinoid system in bull sperm. Theriogenology, 153, 91–101. https://doi.org/10.1016/j.theriogenology.2020.04.031Panzani, D., Rota, A., Tesi, M., Fanelli, D., & Camillo, F. (2018). Update on Donkey Embryo Transfer and Cryopreservation. Journal of Equine Veterinary Science, 65, 50–54. https://doi.org/10.1016/J.JEVS.2017.11.012Patrat, C., Serres, C., & Jouannet, P. (2000). The acrosome reaction in human spermatozoa. Biology of the Cell, 92(3–4), 255–266. https://doi.org/10.1016/s0248-4900(00)01072-8Peña-Alfaro, C. E., Barros, L. O., Carneiro, G. F., Gastal, M. O., & Gastal, E. L. (2014). Embryo transfer in Pega donkeys (Equus asinus) in Brazil. Journal of Equine Veterinary Science, 34(1), 185. https://doi.org/10.1016/J.JEVS.2013.10.135Perez-Osorio, J., Mello, F. G., Juliani, G., Lagares, M., Lago, L., & Henry, M. (2008). Effect on post-thaw viability of equine sperm using stepwise addition of dimethyl formamide and varying cooling and freezing procedures. Animal Reproduction, 5(3–4), 103–109.Pizarro, E., Restrepo, G., Echeverry, J., & Rojano, B. (2013). Effect of seminal plasma on the redox state of cryopreserved stallion semen. Revista MVZ Córdoba, 18, 3672–3680.Quaresma, M., Martins, A. M. F., Rodrigues, J. B., Colaço, J., & Payan-Carreira, R. (2014). Pedigree and herd characterization of a donkey breed vulnerable to extinction. Animal, 8(3), 354–359. https://doi.org/10.1017/S1751731113002218Rathi, R., Colenbrander, B., Bevers, M. M., & Gadella, B. M. (2001). Evaluation of in vitro capacitation of stallion spermatozoa. Biology of Reproduction, 65(2), 462–470. https://doi.org/10.1095/biolreprod65.2.462Restrepo, G., Cantero Naclares, J. M., & Montoya Paez, J. D. (2016). Efecto De La Centrifugación Sobre La Integridad Y La Funcionalidad De Espermatozoides Equinos. Biotecnoloía En El Sector Agropecuario y Agroindustrial, 14(1), 119. https://doi.org/10.18684/bsaa(14)119-125Restrepo, G., Pérez, Daniel., Acosta, Mariano., Camacho, C., & Pérez, J. (2017). Freezing of Equine Semen Under Two Schemes of Addition of Dimethylformamide. Revista de Investigaciones Veterinarias Del Peru, 28(4), 918–927. https://doi.org/10.15381/rivep.v28i4.13884Restrepo, G., Zapata Acosta, K., & Rojano, B. (2015). Evaluación de la capacidad antioxidante total del plasma seminal equino. Zootecnia Tropical, 33(1), 79–87.Ribeiro-Peres, A., Munita-Barbosa, L., Yumi-Kanazawa, M., Mello-Martins, M. I., & Ferreira De Souza, F. (2014). Criopreservación de espermatozoides bovinos extraídos de la cola del epidídimo utilizando los métodos convencional y automatizado. Archivos de Medicina Veterinaria, 46(1), 31–38. https://doi.org/10.4067/S0301-732X2014000100005Rodriguez, P. C., Satorre, M. M., & Beconi, M. T. (2012). Effect of two intracellular calcium modulators on sperm motility and heparin-induced capacitation in cryopreserved bovine spermatozoa. Animal Reproduction Science, 131(3–4), 135–142. https://doi.org/10.1016/j.anireprosci.2012.03.015Rodríguez-Villamil, P., Mentz, D., Ongaratto, F. L., Aguiar, L. H., Rodrigues, J. L., Bertolini, M., & Moura, A. A. (2020). Assessment of binder of sperm protein 1 (BSP1) and heparin effects on in vitro capacitation and fertilization of bovine ejaculated and epididymal sperm. Zygote, 28(6), 489–494. https://doi.org/10.1017/S0967199420000374Rossel, S., Marshall, F., Peters, J., Pilgram, T., Adams, M. D., & O’Connor, D. (2008). Domestication of the donkey: Timing, processes, and indicators. Proceedings of the National Academy of Sciences of the United States of America, 105(10), 3715–3720. https://doi.org/10.1073/pnas.0709692105Rota, A., Sabatini, C., Przybył, A., Ciaramelli, A., Panzani, D., & Camillo, F. (2019). Post-thaw Addition of Caffeine and/or Pentoxifylline Affect Differently Motility of Horse and Donkey-Cryopreserved Spermatozoa. Journal of Equine Veterinary Science, 75, 41–47. https://doi.org/10.1016/j.jevs.2019.01.003Sabatini, C., Mari, G., Mislei, B., Love, C. C., Panzani, D., Camillo, F., & Rota, A. (2014). Effect of Post-Thaw Addition of Seminal Plasma on Motility, Viability and Chromatin Integrity of Cryopreserved Donkey Jack (Equus asinus) Spermatozoa. Reproduction in Domestic Animals, 49(6), 989–994. https://doi.org/https://doi.org/10.1111/rda.12419Sati, L., Cayli, S., Delpiano, E., Sakkas, D., & Huszar, G. (2014). The pattern of tyrosine phosphorylation in human sperm in response to binding to zona pellucida or hyaluronic acid. Reproductive Sciences, 21(5), 573–581. https://doi.org/10.1177/1933719113504467Shahsavari, M. H., Moghaddam, G., Kia, H. D., & Rodrigues, A. P. R. (2019). Effects of new synthetic cryoprotectant agents on histological characteristics of various classes of vitrified bovine pre-antral follicles. Veterinary Research Forum, 10(1), 9–16. https://doi.org/10.30466/vrf.2019.34306Shen, L., Chen, H., Zhu, Q., Wang, Y., Wang, S., Qian, J., Wang, Y., & Qu, H. (2016). Identification of bioactive ingredients with immuno-enhancement and anti-oxidative effects from Fufang-Ejiao-Syrup by LC-MSn combined with bioassays. Journal of Pharmaceutical and Biomedical Analysis, 117, 363–371. https://doi.org/10.1016/j.jpba.2015.09.024Sinha, B., Semmler, J., Eisenhut, T., Eigler, A., & Endres, S. (1995). Enhanced tumor necrosis factor suppression and cyclic adenosine monophosphate accumulation by combination of phosphodiesterase inhibitors and prostanoids. European Journal of Immunology, 25(1), 147–153. https://doi.org/https://doi.org/10.1002/eji.1830250125Sjunnesson, Y. (2020). In vitro fertilisation in domestic mammals—a brief overview. Upsala Journal of Medical Sciences, 125(2), 68–76. https://doi.org/10.1080/03009734.2019.1697911Slanina, T., Miškeje, M., Tirpák, F., Błaszczyk, M., Formicki, G., & Massányi, P. (2018). Caffeine strongly improves motility parameters of Turkey spermatozoa with no effect on cell viability. Acta Veterinaria Hungarica, 66(1), 137–150. https://doi.org/10.1556/004.2018.013Smits, K., Hoogewijs, M., Woelders, H., Daels, P., & Van Soom, A. (2012). Breeding or assisted reproduction? Relevance of the horse model applied to the conservation of endangered equids. Reproduction in Domestic Animals, 47(SUPPL.4), 239–248. https://doi.org/10.1111/j.1439-0531.2012.02082.xSoni, Y., Talluri, T. R., Kumar, A., Ravi, S. K., Mehta, J. S., & Tripathi, B. N. (2019). Effects of different concentration and combinations of cryoprotectants on sperm quality, functional integrity in three Indian horse breeds. Cryobiology, 86(August 2018), 52–57. https://doi.org/10.1016/j.cryobiol.2018.12.005Stanic, P., Sonicki, Z., & Suchanek, E. (2002). Effect of pentoxifylline on motility and membrane integrity of cryopreserved human spermatozoa. International Journal of Andrology, 25(3), 186–190. https://doi.org/10.1046/j.1365-2605.2002.00348.xStephens, T. D., Brooks, R. M., Carrington, J. L., Cheng, L., Carrington, A. C., Porr, C. A., & Splan, R. K. (2013). Effects of pentoxifylline, caffeine, and taurine on post-thaw motility and longevity of equine frozen semen. Journal of Equine Veterinary Science, 33(8), 615–621. https://doi.org/10.1016/j.jevs.2012.10.004Strohmeyer, N., Bharadwaj, M., Costell, M., Fässler, R., & Müller, D. J. (2017). Fibronectin-bound α5β1 integrins sense load and signal to reinforce adhesion in less than a second. Nature Materials, 16(12), 1262–1270. https://doi.org/10.1038/nmat5023Talukdar, D., Ahmed, K., Deori, S., & Das, G. C. (2015). Heparin-induced in vitro capacitation changes of swamp buffalo spermatozoa. Turkish Journal of Veterinary and Animal Sciences, 39(5), 629–633. https://doi.org/10.3906/vet-1501-16Tash, J. S., & Means, A. R. (1982). Regulation of protein phosphorylation and motility of sperm by cyclic adenosine monophosphate and calcium. Biology of Reproduction, 26(4), 745–763. https://doi.org/10.1095/biolreprod26.4.745The Donkey Sanctuary. (2017). Bajo la Piel: El comercio emergente de las pieles de burro y sus consecuencias para el bienestar y la subsistencia de estos animales.Thomas, A. D., Meyers, S. A., & Ball, B. A. (2006). Capacitation-like changes in equine spermatozoa following cryopreservation. Theriogenology, 65(8), 1531–1550. https://doi.org/10.1016/j.theriogenology.2005.08.022Turley, E. A., Noble, P. W., & Bourguignon, L. Y. W. (2002). Signaling properties of hyaluronan receptors. In Journal of Biological Chemistry (Vol. 277, Issue 7, pp. 4589–4592). https://doi.org/10.1074/jbc.R100038200Úsuga, A., Rojano, B. A., & Restrepo, G. (2018). Efecto de los componentes del plasma seminal en la calidad del semen equino. Revista CES Medicina Veterinaria y Zootecnia, 13, 281.Varela, E., Rojas, M., & Restrepo, G. (2020). Membrane stability and mitochondrial activity of bovine sperm frozen with low-density lipoproteins and trehalose. Reproduction in Domestic Animals = Zuchthygiene, 55(2), 146–153. https://doi.org/10.1111/rda.13599Verma, A. S., Agrahari, S., Rastogi, S., & Singh, A. (2011). Biotechnology in the realm of history. In Journal of Pharmacy and Bioallied Sciences (Vol. 3, Issue 3, pp. 321–323). Wolters Kluwer -- Medknow Publications. https://doi.org/10.4103/0975-7406.84430Vidament, M., Vincent, P., Martin, F. X., Magistrini, M., & Blesbois, E. (2009). Differences in ability of jennies and mares to conceive with cooled and frozen semen containing glycerol or not. Animal Reproduction Science, 112(1–2), 22–35. https://doi.org/10.1016/j.anireprosci.2008.03.016Visconti, P. E. (2009). Understanding the molecular basis of sperm capacitation through kinase design. In Proceedings of the National Academy of Sciences of the United States of America (Vol. 106, Issue 3, pp. 667–668). National Academy of Sciences. https://doi.org/10.1073/pnas.0811895106Warren, L. K. (2009). Feeding the Stallion. Agri-Facts, January, 1–4.White, I. G. (1993). Lipids and calcium uptake of sperm in relation to cold shock and preservation: a review. Reproduction, Fertility, and Development, 5(6), 639–658. https://doi.org/10.1071/rd9930639Yilmaz, O., Boztepe, S., & Ertugrul, M. (2012). Domesticated Donkeys – Part II: Types and Breeds. Canadian Journal of Animal Science, 2012(April).Yovich, J. L. (1993). Pentoxifylline: actions and applications in assisted reproduction. Human Reproduction, 8(11), 1786–1791. https://doi.org/10.1093/oxfordjournals.humrep.a137935Zamzami, N., Susin, S. A., Marchetti, P., Hirsch, T., Gómez-Monterrey, I., Castedo, M., & Kroemer, G. (1996). Mitochondrial control of nuclear apoptosis. Journal of Experimental Medicine, 183(4), 1533–1544. https://doi.org/10.1084/jem.183.4.1533Zhao, Q., Tao, C., Pan, J., Wei, Q., Zhu, Z., Wang, L., Liu, M., Huang, J., Yu, F., Chen, X., Zhang, L., & Li, J. (2021). Equine chorionic gonadotropin pretreatment 15 days before fixed-time artificial insemination improves the reproductive performance of replacement gilts. Animal, 15(12), 100406. https://doi.org/10.1016/J.ANIMAL.2021.100406Zheng, C., Jian-tao, Z., Hai-feng, L., & Hong-bin, W. (2016). Laparoscopic Embryo Transfer in Pigs: Knowledge for Surgical Procedures. Journal of Northeast Agricultural University (English Edition), 23(2), 52–58. https://doi.org/10.1016/S1006-8104(16)30047-2BibliotecariosLICENSElicense.txtlicense.txttext/plain; charset=utf-85879https://repositorio.unal.edu.co/bitstream/unal/83291/1/license.txteb34b1cf90b7e1103fc9dfd26be24b4aMD51ORIGINALTesis Burros Luis Mendoza Arenas.pdfTesis Burros Luis Mendoza Arenas.pdfTesis de Maestría en Ciencia - Biotecnologíaapplication/pdf604059https://repositorio.unal.edu.co/bitstream/unal/83291/2/Tesis%20Burros%20Luis%20Mendoza%20Arenas.pdf0d23318ae0d8223fa1ee53914faa5139MD52THUMBNAILTesis Burros Luis Mendoza Arenas.pdf.jpgTesis Burros Luis Mendoza Arenas.pdf.jpgGenerated Thumbnailimage/jpeg3624https://repositorio.unal.edu.co/bitstream/unal/83291/3/Tesis%20Burros%20Luis%20Mendoza%20Arenas.pdf.jpge15f79b65d0c97fc6ed0350e009a6f2cMD53unal/83291oai:repositorio.unal.edu.co:unal/832912023-08-15 23:04:02.736Repositorio Institucional Universidad Nacional de Colombiarepositorio_nal@unal.edu.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 |