English
French
Short-term preservation of canine sperm-binding ability and other metrics using the INRA-96 in comparison to Tris-egg yolk extender.
Abstract: The use of assisted reproductive techniques, such as chilled semen, contributes to the maintenance and genetic improvement of canine breeding. The INRA-96 extender is a commercially available, chemically defined medium that was initially developed for the preservation of equine semen and exhibits preservation potential in the canine species. This research aims to evaluate the INRA-96 extender as an alternative for the short-term preservation of canine semen in terms of sperm quality parameters such as motility and kinetic parameters, integrity and functionality of the plasma membrane in fresh and chilled-rewarmed samples, as well as the sperm-binding ability using the perivitelline membrane of the chicken egg as an indicator of the fertilizing capacity of the preserved semen. A total of 18 ejaculates from 9 French bulldogs (two ejaculates per dog) were collected and divided into two aliquots that were diluted in Tris-egg yolk 20% (control) or INRA-96 to a final concentration of 100 × 10 sperm/mL. Samples were refrigerated in a biological incubator at 5°C and evaluated at 0, 24 and 48 h time points. Comparing the two treatments after 48 h of refrigeration, both extenders showed similar values (p < .5) for the majority of kinetic parameters, with the INRA-96 group promoting a total motility of 88.1 ± 2.9%. In addition, the morphology, integrity and functionality of the plasma membrane were preserved above 70% in this group. Dilution with INRA-96 also provided a significantly higher amount of sperm bound (256.2 ± 21.1) to the perivitelline membrane of the egg yolk compared to the sperm-binding rates (p < .05) achieved at the use of Tris-egg yolk (215.2 ± 21 bound spermatozoa) at 48 h. Our study proved similar functional properties of dog sperm cells treated with INRA-96 in comparison to commonly used home-made Tris-based extender during short-time storage.
© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.
Publication Date: 2023-08-06 PubMed ID: 37545097DOI: 10.1111/rda.14448Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research article examines the use of a commercially available sperm preservation medium, INRA-96, for maintaining canine sperm quality during short-term storage. The findings suggest that INRA-96 is as effective as the traditional Tris-egg yolk extender in preserving important sperm parameters such as motility and plasma membrane integrity.
Study Objective and Method
- The study aimed to evaluate the efficacy of a commercially available semen extender, named INRA-96, in preserving the quality and functionality of canine sperm during short-term storage. The researchers looked at various parameters, including sperm motility, the integrity and functionality of the sperm’s plasma membrane, and the sperm’s binding ability to the perivitelline membrane of an egg.
- 18 semen samples were collected from 9 French Bulldogs, with two samples taken per dog. The samples were divided and diluted with either INRA-96 or Tris-egg yolk 20% (control) to reach a final concentration of 100 x 10 sperm/mL.
- The samples were then refrigerated in a biological incubator at 5°C and evaluated at 0, 24, and 48 hours.
Findings and Observations
- Two days after refrigeration, the two extenders showed similar values for most kinetic parameters. Moreover, the INRA-96 group promoted a total sperm motility of 88.1 ± 2.9%.
- In the samples diluted with INRA-96, the morphology, integrity, and functionality of the plasma membrane were preserved above 70%.
- The INRA-96 extender also resulted in a significantly higher number of sperm binding to the perivitelline membrane of the egg yolk when compared to the control group. Specifically, 256.2 ± 21.1 spermatozoa were found bound to the egg yolk membrane in the INRA-96 group, compared to 215.2 ± 21 in the control group at 48 hours. This is an important finding, as it suggests that INRA-96 may be effective in preserving the fertilising capacity of stored canine sperm.
Conclusion
- The researchers concluded that INRA-96 exhibits similar effects on dog sperm’s functional properties as the commonly used Tris-based extender during short-term storage.
- This finding offers potential for INRA-96’s usage in assisted canine reproductive techniques, contributing to better maintenance and genetic improvement of breeding.
Cite This Article
APA
Cavalcanti TP, Pereira AG, Bezerra LGP, Moreira SSJ, da Silva AM, Matos YG, Jorge-Neto PN, Silva AR.
(2023).
Short-term preservation of canine sperm-binding ability and other metrics using the INRA-96 in comparison to Tris-egg yolk extender.
Reprod Domest Anim, 58(9), 1320-1329.
https://doi.org/10.1111/rda.14448 Publication
Researcher Affiliations
- Universidade de Fortaleza, Fortaleza, Brazil.
- Laboratu00f3rio de Conservau00e7u00e3o de Germoplasma Animal, Universidade Federal Rural do Semi-u00c1rido, Mossoru00f3, Brazil.
- Laboratu00f3rio de Conservau00e7u00e3o de Germoplasma Animal, Universidade Federal Rural do Semi-u00c1rido, Mossoru00f3, Brazil.
- Laboratu00f3rio de Conservau00e7u00e3o de Germoplasma Animal, Universidade Federal Rural do Semi-u00c1rido, Mossoru00f3, Brazil.
- Laboratu00f3rio de Conservau00e7u00e3o de Germoplasma Animal, Universidade Federal Rural do Semi-u00c1rido, Mossoru00f3, Brazil.
- Laboratu00f3rio de Conservau00e7u00e3o de Germoplasma Animal, Universidade Federal Rural do Semi-u00c1rido, Mossoru00f3, Brazil.
- Instituto Reprocon, Campo Grande, Brazil.
- Laboratu00f3rio de Conservau00e7u00e3o de Germoplasma Animal, Universidade Federal Rural do Semi-u00c1rido, Mossoru00f3, Brazil.
MeSH Terms
- Animals
- Dogs
- Male
- Horses
- Semen
- Egg Yolk / chemistry
- Benchmarking
- Sperm Motility
- Semen Preservation / veterinary
- Semen Preservation / methods
- Spermatozoa
- Cryopreservation / veterinary
- Cryopreservation / methods
- Cryoprotective Agents / pharmacology
Grant Funding
- Conselho Nacional de Desenvolvimento Cientu00edfico e Tecnolu00f3gico
- Coordenau00e7u00e3o de Aperfeiu00e7oamento de Pessoal de Nu00edvel Superior
References
This article includes 41 references
- Andersen, A. H., Thinnesen, M., Failing, K., & Goericke-Pesch, S. (2018). Effect of reduced glutathione (GSH) supplementation to Tris-egg yolk extender on chilled semen variables of dogs. Animal Reproduction Science, 198, 145-153. https://doi.org/10.1016/j.anireprosci.2018.09.013
- Arau00fajo, M. S., de Oliveira, H. P. O. L., Paulini, F., Angrimani, D. S. R., Tsunemi, M. H., Dell'Aqua, C. P. F., Pap, F. O., & Souza, F. F. (2022). Seminal plasma does not influence canine semen stored at 5u00b0C for long-term conservation. Biopreservation and Biobanking, 20, 149-162. https://doi.org/10.1089/bio.2021.0054
- Arruda, R. P. (2000). Avaliau00e7u00e3o dos efeitos de diluidores e crioprotetores para o espermatozoide equino pelo uso de microscopia de epifluorescu00eancia, citometria de fluxo, anu00e1lises computadorizadas da motilidade (CASA) e da morfometria (ASMA). [Evaluation of the effects of extenders and cryoprotectants on equine sperm using epifluorescence microscopy, flow cytometry, computerized motility analysis (CASA) and morphometry (ASMA)] (Habilitation thesis). Universidade de Su00e3o Paulo, Su00e3o Paulo, p. 121.
- Bannasch, D., Famula, T., Donner, J., Anderson, H., Honkanen, L., Batcher, K., Safra, N., Thomasy, S., & Rebhun, R. (2021). The effect of inbreeding, body size and morphology on health in dog breeds. Canine Medicine and Genetics, 8, 12. https://doi.org/10.1186/s40575-021-00111-4
- Barbosa, B., Silva, H. V. R., Tabosa, B., Nunes, T., Magalhu00e3es, F. F., & Silva, L. D. M. (2019). Morphological and morphometric characterization of domestic cat epididymal sperm. Reproduction in Domestic Animals, 54, 1630-1636. https://doi.org/10.1111/rda.13572
- Batellier, F., Vidament, M., Fauquant, J., Duchamp, G., Arnaud, G., Yvon, J. M., & Magistrini, M. (2001). Advances in cooled semen technology. Animal Reproduction Science, 68, 181-190. https://doi.org/10.1016/s0378-4320(01)00155-5
- Bencharif, D., Amirat-Briand, L., Guillou, J. L. E., Vialtelle, C., Anton, M., Schmitt, E., Desherces, S., Barriere, P., & Tainturier, D. (2013). Refrigeration of canine sperm at +4u00b0C: Comparative study of four different extenders for the refrigeration of canine sperm at +4u00b0C: LDL, Tris egg yolk, Equexu00ae, and INRA96u00ae. Revue de Mu00e9decine Veterinaire, 164, 252-262.
- Bousseau, S., Brillard, J. P., Guienne, B. M., Guu00e9rin, B., Camus, A., & Lechat, M. (1998). Comparision of bacteriological qualities of various egg yolk sources and the in vitro and in vivo fertilizing potential of bovine semen frozen in egg yolk or lecithin-based diluents. Theriogenology, 50, 699-706. https://doi.org/10.1016/s0093-691x(98)00175-7
- Brito, M. M., Losano, J. D. A., Angrimani, D. S. R., Lu00facio, C. F., Dalmazzo, A., Nichi, M., & Vannuchi, C. I. (2017). Sperm-binding to the perivitelline membrane of chicken egg yolk as a functional test for sperm evaluation in dogs. Brazilian Journal of Veterinary Research and Animal Science, 54, 366-374. https://doi.org/10.11606/issn.1678-4456.bjvras.2017.127331
- Campos, L. B., Peixoto, G. C. X., Silva, A. M., Souza, A. L. P., Castelo, T. S., Maia, K. M., Pereira, A. F., & Silva, A. R. (2017). Estimating the binding ability of collared peccary (Pecari tajacu Linnaeus, 1758) sperm using heterologous substrates. Theriogenology, 92, 57-62. https://doi.org/10.1016/j.theriogenology.2017.01.008
- Cardoso, J. F. S., Paula, N. R. O., Uchoa, D. C., & Silva, L. D. M. (2010). Different egg yolk concentrations on the quality of the canine semen diluted in ACP-106 and chilled under 4u00b0C. Comunicata Scientiae, 1, 146-152.
- Cardoso, R. C. S., Silva, A. R., & Silva, L. D. M. (2006). Comparison of two dilution rates on canine semen quality after cryopreservation in a coconut water extender. Animal Reproduction Science, 92, 384-391. https://doi.org/10.1016/j.anireprosci.2005.05.028
- CBRA - Colu00e9gio Brasileiro de Reproduu00e7u00e3o Animal. (2013). Manual para exame androlu00f3gico e avaliau00e7u00e3o de su00eamen animal [Manual for andrological examination and evaluation of animal semen] (3rd ed., p. 104). CBRA.
- Chu0142opik, A., & Wysokiu0144ska, A. (2019). Canine spermatozoa - What do we know about their morphology and physiology? An overview. Reproduction in Domestic Animals, 55, 113-126. https://doi.org/10.1111/rda.13596
- Colombo, M., Morseli, M. G., Franchi, G., Schu00e4fer-Somi, S., & Luvoni, G. C. (2022). Freezability of dog semen after collection in field conditions and cooled transport. Animals, 12, 816. https://doi.org/10.3390/ani12070816
- Dorado, J., Gu00e1lvez, M. J., Murabito, M. R., Muu00f1oz-Serrano, A., & Hidalgo, M. (2011). Identification of sperm subpopulations in canine ejaculates: Effects of cold storage and egg yolk concentration. Animal Reproduction Science, 127, 106-113. https://doi.org/10.1016/j.anireprosci.2011.07.005
- Hesser, A., Darr, C., Gonzales, K., Power, H., Scanlan, T., Thompson, J., Love, C., Christensen, B., & Meyers, S. (2017). Semen evaluation and fertility assessment in a purebred dog breeding facility. Theriogenology, 87, 115-123. https://doi.org/10.1016/j.theriogenology.2016.08.012
- Iguer-ouada, M., & Verstegen, J. P. (2001). Evaluation of the u201cHamilton Thron Computer-Based Aumated Systemu201d for dog semen analysis. Theriogenology, 55, 733-749. https://doi.org/10.1016/s0093-691x(01)00440-x
- Johnston, S. D. (2001). Semen collection, evaluation and preservation. In S. D. Johnston, M. V. R. Kustritz, & P. N. S. Olson (Eds.), Canine and feline theriogenology (pp. 287-306). Elsevier.
- Linde-Forsberg, C. (1991). Achieving canine pregnancy by using frozen or chilled extended semen. Veterinary Clinics of North America: Small Animal Practice, 21, 467-485. https://doi.org/10.1016/s0195-5616(91)50054-1
- Linde-Forsberg, C. (2001). Regulations and recommendations for international shipment of chilled and frozen canine semen. In P. W. Concannon, G. England, J. Verstegen, & C. Linde-Forsberg (Eds.), Recent advances in small animal reproduction. International Veterinary Information Service.
- Meng, Y., Qiu, N., Guyonnet, V., Keast, R., Zhu, C., & Mine, Y. (2022). UHPLC-Q-Orbitrap -based untargeted lipidomics reveals the variation of yolk lipids during egg storage. Journal of the Science of Food and Agriculture, 102, 5690-5699. https://doi.org/10.1002/jsfa.11916
- Mota-Filho, A. C., Silva, H. V. R., Nunes, T. G. P., Souza, M. B., Freitas, L. A., Araujo, A. A., & Silva, L. D. M. (2014). Cryopreservation of canine epididymal sperm using ACP-106c and TRIS. Cryobiology, 69, 17-21. https://doi.org/10.1016/j.cryobiol.2014.04.013
- Nothling, J. O., & Schuttleworth, R. (2005). The effect of straw size, freezing rate and thawing rate upon post-thaw quality of dog semen. Theriogenology, 63, 1469-1480. https://doi.org/10.1016/j.theriogenology.2004.07.012
- Novello, G., Podico, G., Segabinazzi, L. G. T. M., Lima, F. S., & Canisso, I. F. (2020). Stallion semen cooling using native phosphocaseinate-based extender and sodium caseinate cholesterol-loaded cyclodextrin-based extender. Journal of Equine Veterinary Science, 92, 103104. https://doi.org/10.1016/j.jevs.2020.103104
- Oettlu00e9, E. E. (1993). Sperm morphology and fertility in the dog. Journal of Reproduction and Fertility - Supplement, 47, 257-260.
- Padilla, A. W., & Foote, R. H. (1991). Extender and centrifugation effects on the motility patterns of slow-cooled stallion spermatozoa. Journal of Animal Science, 69, 3308-3313. https://doi.org/10.2527/1991.6983308x
- Peu00f1a, A. I., Adu00e1n, S., Quintela, L. A., Becerra, J. J., & Herradu00f3n, P. G. (2018). Relationship between motile sperm subpopulations identified in frozen-thawed dog semen samples and their ability to bind to the zona pellucida of canine oocytes. Reproduction in Domestic Animals, 53, 14-22. https://doi.org/10.1111/rda.13349
- Quintela, A. T., Oliveira, I. R. S., Souza, A. O., Gusmu00e3o, A. L., & Silva, A. R. (2010). Water-induced hypo-osmotic test for the evaluation of canine sperm membrane integrity. Animal Reproduction, 7, 70-74.
- Quintero-Moreno, A., Miru00f3, J., Teresa Rigau, A., & Rodriguez-Gil, J. E. (2003). Identification of sperm subpopulations with specific motility characteristics in stallion ejaculates. Theriogenology, 59, 1973-1990. https://doi.org/10.1016/S0093-691X(02)01297-9
- Ramirez, J. P., Carreras, A., & Mendonza, C. (1992). Sperm plasma membrane integrity in fertile and infertile men. Andrologia, 24, 141-144. https://doi.org/10.1111/j.1439-0272.1992.tb02627.x
- Rigau, T., Farru00e9, M., Ballester, J., Mogas, T., Peu00f1a, A., & Rodru00edguez-Gil, J. E. (2001). Effects of glucose and fructose on motility patterns of dog spermatozoa from fresh ejaculates. Theriogenology, 56, 801-815. https://doi.org/10.1016/S0093-691X(01)00609-4
- Robert, M. A., Jayaprakash, G., Pawshe, M., Tamilmani, T., & Sathiyabarathi, M. (2016). Collection and evaluation of canine semen: A review. International Journal of Science, Environment and Technology, 5, 1586-1595.
- Salamon, S., & Maxwell, W. M. C. (2000). Storage of ram semen. Animal Reproduction Science, 62(1-3), 77-111. https://doi.org/10.1016/s0378-4320(00)00155-x
- Silva, A. R., Cardoso, R. C. S., Silva, L. D. M., Chirinu00e9a, V. H., Lopes, M. D., & Souza, F. F. (2006). Prognostic value of canine frozen-thawed semen parameters on in vitro sperm-oocyte interactions. Theriogenology, 66, 456-462. https://doi.org/10.1016/j.theriogenology.2005.12.00
- Silva, A. R., Cardoso, R. C. S., Uchoa, D. C., & Silva, L. D. M. (2002). Effect of tris-buffer, egg yolk and glycerol on canine semen freezing. The Veterinary Journal, 164, 244-246. https://doi.org/10.1053/tvjl.2002.0704
- Stuart, C. C., Vaughan, J. L., Kershaw, C. M., Graaf, S. P., & Bathgate, R. (2019). Effect of diluent type, cryoprotectant concentration, storage method and freeze/thaw rates on the post-thaw quality and fertility of cryopreserved alpaca spermatozoa. Scientific Reports, 9, 12826. https://doi.org/10.1038/s41598-019-49203-z
- Suzuki, H., Watanabe, H., & Abe, Y. (2022). Assisted reproductive techniques for canines: Preservation of genetic material in domestic dogs. Journal of Reproduction and Development, 68, 1-11. https://doi.org/10.1262/jrd.2021-111
- Takeuchi, Y., Nishimura, K., Aoki, N., Adachi, T., Sato, C., Kitajima, K., & Matsuda, T. (1999). A 42-kDa glycoprotein from chicken egg-envelope, an avian homolog of the ZPC family glycoproteins in mammalian zona pellucida: Its first identification, cDNA cloning and granulosa cell-specific expression. European Journal of Biochemistry, 260, 736-742. https://doi.org/10.1046/j.1432-1327.1999.00203.x
- Teixeira, D. O., Silva, H. V. R., Brito, B. F., Barbosa, B. S., Tabosa, B. E. A., & Silva, L. D. M. (2022). Sperm quality and morphometry characterization of cryopreserved canine sperm in ACP-106c or TRIS. Animal Reproduction, 19, e20210069. https://doi.org/10.1590/1984-3143-AR2021-0069
- Zenteno, E. S., Rojano, B., & Betauncur, G. R. (2023). Influence of thawing temperature on sperm motility, structure, and metabolism of frozen bovine semen. Ciu00eancia Rural, 53, e20210731. https://doi.org/10.1590/0103-8478cr20210731
Citations
This article has been cited 0 times.Secure Payments
