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Reproduction in domestic animals = Zuchthygiene2023; 58(9); 1320-1329; doi: 10.1111/rda.14448

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.
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Publication Date: 2023-08-06 PubMed ID: 37545097DOI: 10.1111/rda.14448Google Scholar: Lookup
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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

Reproduction in domestic animals = Zuchthygiene
ISSN: 1439-0531
NlmUniqueID: 9015668
Country: Germany
Language: English
Volume: 58
Issue: 9
Pages: 1320-1329

Researcher Affiliations

Cavalcanti, Thales Pinheiro
  • Universidade de Fortaleza, Fortaleza, Brazil.
Pereira, Ana Glória
  • Laboratório de Conservação de Germoplasma Animal, Universidade Federal Rural do Semi-Árido, Mossoró, Brazil.
Bezerra, Luana Grasiele Pereira
  • Laboratório de Conservação de Germoplasma Animal, Universidade Federal Rural do Semi-Árido, Mossoró, Brazil.
Moreira, Samara Sandy Jerônimo
  • Laboratório de Conservação de Germoplasma Animal, Universidade Federal Rural do Semi-Árido, Mossoró, Brazil.
da Silva, Andreia Maria
  • Laboratório de Conservação de Germoplasma Animal, Universidade Federal Rural do Semi-Árido, Mossoró, Brazil.
Matos, Yuri Gonçalves
  • Laboratório de Conservação de Germoplasma Animal, Universidade Federal Rural do Semi-Árido, Mossoró, Brazil.
Jorge-Neto, Pedro Nacib
  • Instituto Reprocon, Campo Grande, Brazil.
Silva, Alexandre Rodrigues
  • Laboratório de Conservação de Germoplasma Animal, Universidade Federal Rural do Semi-Árido, Mossoró, 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

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Citations

This article has been cited 1 times.
  1. Karimi I, Mohammad LJ, Suvitha A, Haidari Z, Schiöth HB. Comprehensive overview of the toxicities of small-molecule cryoprotectants for carnivorous spermatozoa: foundation for computational cryobiotechnology. Front Toxicol 2025;7:1477822.
    doi: 10.3389/ftox.2025.1477822pubmed: 40078955google scholar: lookup
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