EFECTO DE DOS PROTOCOLOS DE CRIOPRESERVACIÓN DE SEMEN SOBRE LOS  RESULTADOS DE INSEMINACIÓN ARTIFICIAL EQUINA

Carlos Fuentes.; Félix Contreras.; Pacífico Bonilla.; Alex Solís-Corrales.

doi:10.48204/j.ia.v7n2.a7493

Submitted June 18, 2025

Published 2025-06-06

Artículos

Vol. 7 No. 2 (2025): Revista investigaciones agropecuarias

EFFECT OF TWO SEMEN CRYOPRESERVATION PROTOCOLS ON EQUINE ARTIFICIAL INSEMINATION OUTCOMES)

Carlos Fuentes. ⁺⁻
Félix Contreras. ⁺⁻
Pacífico Bonilla. ⁺⁻
Alex Solís-Corrales. ⁺⁻

DOI https://doi.org/10.48204/j.ia.v7n2.a7493

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DOI: 10.48204/j.ia.v7n2.a7493

Published: 2025-06-06

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Fuentes. , C., Contreras. , F., Bonilla. , P., & Solís-Corrales. , A. (2025). EFFECT OF TWO SEMEN CRYOPRESERVATION PROTOCOLS ON EQUINE ARTIFICIAL INSEMINATION OUTCOMES) . Journal of Agricultural Research, 7(2), 70–79. https://doi.org/10.48204/j.ia.v7n2.a7493

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

Abstract

The objective of this study was to evaluate the effect of two semen cryopreservation protocols on equine artificial insemination results. Treatment one (T1) used a centrifugation of 600 g for 11 minutes, while treatment two (T2) used a force of 750 g for eight minutes. Thirteen Quarter Horse mares, aged between four and eight years, were used. The body condition of the mares was approximately six to seven (on a scale of one to nine). Thirteen inseminations were performed with cryopreserved semen (T1: seven inseminations and T2: six inseminations with 750 g semen for eight minutes). The data were arranged in a contingency table. For the statistical analysis, the ChiSquare test was performed, using the IBM SPSS Statisctics version: 29.0.0.0 (241), where significant differences (P<0.05) were found for pregnancy rates when cryopreserved semen was used in T1 over T2 (T1: 71.40% and T2:

16.70%). It is concluded that T1 cryopreserved semen gave the best pregnancy rates using fixed-time artificial insemination.

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Avanzi, B. R., Ramos, R. dos S., Araujo, G. H. M., Fioratti, E. G., Trinca, L. A., Dell’Aqua, J. A., Melo e Oña, C. M., Zahn, F. S., Martin, I., Alvarenga, M. A. y Papa, F. O. (2015). Fixed-time insemination with frozen semen in mares: is it suitable for poorly fertile stallions? Theriogenology, 83(9), 1389-1393. https://doi.org/10.1016/j.theriogenology.2014.07.007
Alonso, A., Baca Castex, C., Pinto, M., Caldevilla, M., Ferrante, A. y Miragaya, M. (2016). Uso de semen congelado en programas de inseminación artificial en equinos. Spermova, 2(6), 107-109. https://doi.org/10.18548/aspe/0004.04
Alvarenga, M. A., Papa, F. O. y Neto, C. R. (2017). Técnicas para incremento da qualidade do sêmen de garanhões. Revista Brasileira de Reprodução Animal, 41(1), 81-85.
Alvarenga, M. A., Papa, F. O. y Ramires Neto, C. (2016). Advances in Stallion Semen Cryopreservation. Veterinary Clinics of North America: Equine Practice, 32(3), 521-530. https://doi.org/10.1016/j.cveq.2016.08.003
Cazales Penino, N., Estradé, M. J., Costa Mattos, R., Cazales Penino, N., Estradé, M. J. y Costa Mattos, R. (2020). Inseminación artificial con semen congelado equino: reacción inflamatoria, transporte espermático y técnica de inseminación. Veterinaria (Montevideo), 56(214). https://doi.org/10.29155/vet.56.214.2
Cerny, K. L., Hughes, S., Campos, J. R., Coleman, R. J., Troedsson, M. H. T. y Squires, E. L. (2012). Fertility of Mares Inseminated With Frozen-Thawed Semen Processed by Single Layer Centrifugation Through a Colloid. Journal of Equine Veterinary Science, 32(5), 289-291. https://doi.org/10.1016/j.jevs.2011.09.075
Contreras, F. A. (2022). Evaluación de dos protocolos de criopreservación de semen equino. [Tesis de Licenciatura]. Universidad de Panamá (Panamá). Facultad de Ciencias Agropecuarias.
Crowe, C. A. M., Ravenhill, P. J., Hepburn, R. J. y Shepherd, C. H. (2008). A retrospective study of artificial insemination of 251 mares using chilled and fixed time frozen-thawed semen. Equine Veterinary Journal, 40(6), 572-576. https://doi.org/10.2746/042516408x281199
Fernández, F., Hernández, J., Rodríguez, S. y Velásquez, H. (2008). Fertilidad en yeguas cuarto de milla tratadas con gonadotropina coriónica humana (hCG) utilizando semen congelado. Revista de Salud Animal, 30(3); 184-188.
Harvey, A. M., Ramp, D. y Mellor, D. J. (2022). Review of the Foundational Knowledge Required for Assessing Horse Welfare. Animals, 12(23), 3385. https://doi.org/10.3390/ani12233385
Immonen, I. y Cuervo-Arango, J. (2020). Effect of Timing of Postovulatory Insemination Relative to Human Chorionic Gonadotropin/Buserelin Treatment With 1 Straw of Frozen-Thawed Semen on Mare Fertility. Journal of Equine Veterinary Science, 87, 102900. https://doi.org/10.1016/j.jevs.2019.102900
Loomis, P. R. (2001). The equine frozen semen industry. Animal Reproduction Science, 68(3-4), 191-200. https://doi.org/10.1016/s0378-4320(01)00156-7
Love, C. C., Blanchard, T. L., Varner, D. D., Brinsko, S. P., Voge, J., Bliss, S., Sudderth, K., Teague, S. y LaCaze, K. (2012). Effect of daily semen centrifugation and resuspension on the longevity of equine sperm quality following cooled storage. Theriogenology, 77(9), 1911-1917. https://doi.org/10.1016/j.theriogenology.2012.01.010
Mari, G., Bucci, D., Love, C. C., Mislei, B., Rizzato, G., Giaretta, E., Merlo, B. y Spinaci, M. (2015). Effect of cushioned or single layer semen centrifugation before sex sorting on frozen stallion semen quality. Theriogenology, 83(6), 953-958. https://doi.org/10.1016/j.theriogenology.2014.11.031
Metcalf, E. S. (2007). The efficient use of equine cryopreserved semen. Theriogenology, 68(3), 423–428. https://doi.org/10.1016/j.theriogenology.2007.04.039
Olguín, R. O. y Esquivel, V. R. (2011). Manual de inseminación artificial para la operatividad de semen congelado en yeguas (Equus caballus) del Criadero Militar de Ganado Santa Gertrudis Chihuahua. Universidad Autónoma Chapingo. Recuperado de https://zootecnia.chapingo.mx/assets/11olguin.pdf
Oliveira, R. A., Rubin, M. I. B. y Silva, C. A. M. (2013). Índice De Prenhez Com Sêmen Congelado De Garanhões Da Raça Crioula Usando Glicerol Ou Dimetilformamida Como Crioprotetores. Ciência Animal Brasileira, 14(4). https://doi.org/10.5216/cab.v14i4.18923
Pillet, E., Batellier, F., Duchamp, G., Furstoss, V., Vern, Y. L., Kerboeuf, D., Vidament, M. y Magistrini, M. (2008). Freezing stallion semen in INRA96®-based extender improves fertility rates in comparison with INRA82. Dairy Science and Technology. 88, 257-265 https://doi.org/10.1051/dst:2008002
Ramires-Neto, C., Monteiro, G. A., Soares, R. F., Pedrazzi, C., Dell'aqua, J. A., Papa, F. O., Castro-Chaves, M. y Alvarenga, M. A. (2013). New seminal plasma removal method for freezing stallion semen. Theriogenology, 79(7), 1120-1123.e1. https://doi.org/10.1016/j.theriogenology.2013.01.014
Restrepo-Betancurt, G., Cantero-Nanclares, J. y Montoya-Páez, J. (2016). Efecto De La Centrifugación Sobre La Integridad Y La Funcionalidad De Espermatozoides Equinos. Biotecnología en el Sector Agropecuario y Agroindustrial, 14(1), 119-125. https://doi.org/10.18684/BSAA(14)119-125
Restrepo-Betancurt, G., Duque Cortés, J. y Montoya-Páez, J. (2012). Effect of two protocols of cryopreservation on fertilizing capacity of stallion (equus caballus) semen / efecto de dos protocolos de criopreservación sobre la capacidad fecundante de semen equino (equus caballus). Universidad Nacional de Colombia - Sede Medellín. Recuperado de https://repositorio.unal.edu.co/bitstream/handle/unal/71546/36504-154209-1-PB.pdf?sequence=1&isAllowed=y
Ribeiro, B., Ramos, R., Araujo, G.H., Fioratti, E. G., Trinca, L.A., Dell´Aqua, J.A., Melo E Oña, C.M., Zahn, F.S., Martin, I., Alvarenga, M.A., y Papa, F.O. (2015). Fixed – time insemination with frozen semen in mares: is it suitable for poorly fertile stallions? Theriogenology, 83(9), 1389 – 1393. https://doi.org/10.1016/j.theriogenology.2014.07.007
Samper, J., Estrada, A. J. y McKinnon, A. O. (2007). Insemination with Frozen Semen. Current Therapy in Equine Reproduction, 285-288. https://doi.org/10.1016/b978-0-7216-0252-3.50049-7
Samper, J. C. (2008). Induction of estrus and ovulation: Why some mares respond and others do not. Theriogenology, 70(3), 445-447. https://doi.org/10.1016/j.theriogenology.2008.04.040
Sánchez, R., Gómez, I. y Samper, J. C. (2009). Artificial Insemination with Frozen Semen. Elsevier EBooks, 175-183. https://doi.org/10.1016/b978-1-4160-5234-0.00015-5
Sharafi, M., Borghei-Rad, S. M., Hezavehei, M., Shahverdi, A. y Benson, J. D. (2022). Cryopreservation of Semen in Domestic Animals: A Review of Current Challenges, Applications, and Prospective Strategies. Animals, 12(23), 3271. https://doi.org/10.3390/ani12233271
Sielhorst, J., Hagen, C., Behrendt, D., Schuette, B., Burger, D., Martinsson, G. y Sieme, H. (2016). Effect of Multiple Freezing of Stallion Semen on Sperm Quality and Fertility. Journal of Equine Veterinary Science, 40, 56-61. https://doi.org/10.1016/j.jevs.2016.01.014
Vidament, M., Dupere, A. M., Julienne, P., Evain, A., Noue, P. y Palmer, E. (1997). Equine frozen semen: Freezability and fertility field results. Theriogenology, 48(6), 907-917. https://doi.org/10.1016/s0093-691x(97)00319-1

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[1] Avanzi, B. R., Ramos, R. dos S., Araujo, G. H. M., Fioratti, E. G., Trinca, L. A., Dell’Aqua, J. A., Melo e Oña, C. M., Zahn, F. S., Martin, I., Alvarenga, M. A. y Papa, F. O. (2015). Fixed-time insemination with frozen semen in mares: is it suitable for poorly fertile stallions? Theriogenology, 83(9), 1389-1393. https://doi.org/10.1016/j.theriogenology.2014.07.007

[2] Alonso, A., Baca Castex, C., Pinto, M., Caldevilla, M., Ferrante, A. y Miragaya, M. (2016). Uso de semen congelado en programas de inseminación artificial en equinos. Spermova, 2(6), 107-109. https://doi.org/10.18548/aspe/0004.04

[3] Alvarenga, M. A., Papa, F. O. y Neto, C. R. (2017). Técnicas para incremento da qualidade do sêmen de garanhões. Revista Brasileira de Reprodução Animal, 41(1), 81-85.

[4] Alvarenga, M. A., Papa, F. O. y Ramires Neto, C. (2016). Advances in Stallion Semen Cryopreservation. Veterinary Clinics of North America: Equine Practice, 32(3), 521-530. https://doi.org/10.1016/j.cveq.2016.08.003

[5] Cazales Penino, N., Estradé, M. J., Costa Mattos, R., Cazales Penino, N., Estradé, M. J. y Costa Mattos, R. (2020). Inseminación artificial con semen congelado equino: reacción inflamatoria, transporte espermático y técnica de inseminación. Veterinaria (Montevideo), 56(214). https://doi.org/10.29155/vet.56.214.2

[6] Cerny, K. L., Hughes, S., Campos, J. R., Coleman, R. J., Troedsson, M. H. T. y Squires, E. L. (2012). Fertility of Mares Inseminated With Frozen-Thawed Semen Processed by Single Layer Centrifugation Through a Colloid. Journal of Equine Veterinary Science, 32(5), 289-291. https://doi.org/10.1016/j.jevs.2011.09.075

[7] Contreras, F. A. (2022). Evaluación de dos protocolos de criopreservación de semen equino. [Tesis de Licenciatura]. Universidad de Panamá (Panamá). Facultad de Ciencias Agropecuarias.

[8] Crowe, C. A. M., Ravenhill, P. J., Hepburn, R. J. y Shepherd, C. H. (2008). A retrospective study of artificial insemination of 251 mares using chilled and fixed time frozen-thawed semen. Equine Veterinary Journal, 40(6), 572-576. https://doi.org/10.2746/042516408x281199

[9] Fernández, F., Hernández, J., Rodríguez, S. y Velásquez, H. (2008). Fertilidad en yeguas cuarto de milla tratadas con gonadotropina coriónica humana (hCG) utilizando semen congelado. Revista de Salud Animal, 30(3); 184-188.

[10] Harvey, A. M., Ramp, D. y Mellor, D. J. (2022). Review of the Foundational Knowledge Required for Assessing Horse Welfare. Animals, 12(23), 3385. https://doi.org/10.3390/ani12233385

[11] Immonen, I. y Cuervo-Arango, J. (2020). Effect of Timing of Postovulatory Insemination Relative to Human Chorionic Gonadotropin/Buserelin Treatment With 1 Straw of Frozen-Thawed Semen on Mare Fertility. Journal of Equine Veterinary Science, 87, 102900. https://doi.org/10.1016/j.jevs.2019.102900

[12] Loomis, P. R. (2001). The equine frozen semen industry. Animal Reproduction Science, 68(3-4), 191-200. https://doi.org/10.1016/s0378-4320(01)00156-7

[13] Love, C. C., Blanchard, T. L., Varner, D. D., Brinsko, S. P., Voge, J., Bliss, S., Sudderth, K., Teague, S. y LaCaze, K. (2012). Effect of daily semen centrifugation and resuspension on the longevity of equine sperm quality following cooled storage. Theriogenology, 77(9), 1911-1917. https://doi.org/10.1016/j.theriogenology.2012.01.010

[14] Mari, G., Bucci, D., Love, C. C., Mislei, B., Rizzato, G., Giaretta, E., Merlo, B. y Spinaci, M. (2015). Effect of cushioned or single layer semen centrifugation before sex sorting on frozen stallion semen quality. Theriogenology, 83(6), 953-958. https://doi.org/10.1016/j.theriogenology.2014.11.031

[15] Metcalf, E. S. (2007). The efficient use of equine cryopreserved semen. Theriogenology, 68(3), 423–428. https://doi.org/10.1016/j.theriogenology.2007.04.039

[16] Olguín, R. O. y Esquivel, V. R. (2011). Manual de inseminación artificial para la operatividad de semen congelado en yeguas (Equus caballus) del Criadero Militar de Ganado Santa Gertrudis Chihuahua. Universidad Autónoma Chapingo. Recuperado de https://zootecnia.chapingo.mx/assets/11olguin.pdf

[17] Oliveira, R. A., Rubin, M. I. B. y Silva, C. A. M. (2013). Índice De Prenhez Com Sêmen Congelado De Garanhões Da Raça Crioula Usando Glicerol Ou Dimetilformamida Como Crioprotetores. Ciência Animal Brasileira, 14(4). https://doi.org/10.5216/cab.v14i4.18923

[18] Pillet, E., Batellier, F., Duchamp, G., Furstoss, V., Vern, Y. L., Kerboeuf, D., Vidament, M. y Magistrini, M. (2008). Freezing stallion semen in INRA96®-based extender improves fertility rates in comparison with INRA82. Dairy Science and Technology. 88, 257-265 https://doi.org/10.1051/dst:2008002

[19] Ramires-Neto, C., Monteiro, G. A., Soares, R. F., Pedrazzi, C., Dell'aqua, J. A., Papa, F. O., Castro-Chaves, M. y Alvarenga, M. A. (2013). New seminal plasma removal method for freezing stallion semen. Theriogenology, 79(7), 1120-1123.e1. https://doi.org/10.1016/j.theriogenology.2013.01.014

[20] Restrepo-Betancurt, G., Cantero-Nanclares, J. y Montoya-Páez, J. (2016). Efecto De La Centrifugación Sobre La Integridad Y La Funcionalidad De Espermatozoides Equinos. Biotecnología en el Sector Agropecuario y Agroindustrial, 14(1), 119-125. https://doi.org/10.18684/BSAA(14)119-125

[21] Restrepo-Betancurt, G., Duque Cortés, J. y Montoya-Páez, J. (2012). Effect of two protocols of cryopreservation on fertilizing capacity of stallion (equus caballus) semen / efecto de dos protocolos de criopreservación sobre la capacidad fecundante de semen equino (equus caballus). Universidad Nacional de Colombia - Sede Medellín. Recuperado de https://repositorio.unal.edu.co/bitstream/handle/unal/71546/36504-154209-1-PB.pdf?sequence=1&isAllowed=y

[22] Ribeiro, B., Ramos, R., Araujo, G.H., Fioratti, E. G., Trinca, L.A., Dell´Aqua, J.A., Melo E Oña, C.M., Zahn, F.S., Martin, I., Alvarenga, M.A., y Papa, F.O. (2015). Fixed – time insemination with frozen semen in mares: is it suitable for poorly fertile stallions? Theriogenology, 83(9), 1389 – 1393. https://doi.org/10.1016/j.theriogenology.2014.07.007

[23] Samper, J., Estrada, A. J. y McKinnon, A. O. (2007). Insemination with Frozen Semen. Current Therapy in Equine Reproduction, 285-288. https://doi.org/10.1016/b978-0-7216-0252-3.50049-7

[24] Samper, J. C. (2008). Induction of estrus and ovulation: Why some mares respond and others do not. Theriogenology, 70(3), 445-447. https://doi.org/10.1016/j.theriogenology.2008.04.040

[25] Sánchez, R., Gómez, I. y Samper, J. C. (2009). Artificial Insemination with Frozen Semen. Elsevier EBooks, 175-183. https://doi.org/10.1016/b978-1-4160-5234-0.00015-5

[26] Sharafi, M., Borghei-Rad, S. M., Hezavehei, M., Shahverdi, A. y Benson, J. D. (2022). Cryopreservation of Semen in Domestic Animals: A Review of Current Challenges, Applications, and Prospective Strategies. Animals, 12(23), 3271. https://doi.org/10.3390/ani12233271

[27] Sielhorst, J., Hagen, C., Behrendt, D., Schuette, B., Burger, D., Martinsson, G. y Sieme, H. (2016). Effect of Multiple Freezing of Stallion Semen on Sperm Quality and Fertility. Journal of Equine Veterinary Science, 40, 56-61. https://doi.org/10.1016/j.jevs.2016.01.014

[28] Vidament, M., Dupere, A. M., Julienne, P., Evain, A., Noue, P. y Palmer, E. (1997). Equine frozen semen: Freezability and fertility field results. Theriogenology, 48(6), 907-917. https://doi.org/10.1016/s0093-691x(97)00319-1

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