Delayed insemination is successful with a new extender for storing fresh equine semen at 15 degrees C under aerobic conditions.
Abstract: Milk-based semen diluents are known to be practical and effective in protecting equine spermatozoa during storage before artificial insemination. Milk is a biological fluid with a complex composition and contains components which are beneficial or harmful to spermatozoa. The aim of this study was to test the fertility of stallion semen after long-term storage using different milk diluents (INRA 82 or Kenney's diluent) vs one diluent chemically defined (INRA 96), which is composed of efficient milk components and optimized for sperm survival and storage temperature. The milk fraction used was that which best maintained spermatozoal survival based on motility measured in previous studies. Four breeding trials were conducted to determine the influence of combination of new diluent and storage conditions on fertility of the stallion. We compared the standard protocol of storing semen in a skim milk diluent (INRA 82 or Kenney's diluent) at 4 degrees C under anaerobic conditions with the experimental protocol which consisted of storing in a chemically defined, milk-free diluent (INRA 96), at 15 degrees C, under aerobic conditions. After 4 breeding trials, in which the semen was stored for 24 h under the 2 protocols, we obtained 57% (n = 178) and 40% (n = 173) of fertility per cycle using the experimental and the standard protocol respectively (p < 0.001). Another breeding trial was conducted to determine the influence of storage time on the fertility of spermatozoa. We have compared the fertility of semen inseminated immediately (68% of fertility per cycle, n = 50) vs the fertility of semen stored under the experimental protocol for 72 h before insemination (48% of fertility per cycle, n = 52). The experimental protocol improved sperm fertility compared to the standard protocol and seems to be a particular alternative for stallions with cold shock sensitive spermatozoa. Storing semen for 72 h under the experimental protocol seems to be useful in the field.
Publication Date: 2000-03-29 PubMed ID: 10734490DOI: 10.1016/s0093-691x(98)00130-7Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research examines the impact of using different diluents for storing stallion semen to optimize fertility in delayed insemination. Findings suggest that using a chemically defined, milk-free diluent (INRA 96) at 15 degrees Celsius under aerobic conditions improves fertility rates compared to the standard protocol of storing semen in a skim milk-based diluent at 4 degrees Celsius under anaerobic conditions.
Objective and Methodology
- The objective of the research was to explore the fertility of stallion semen after long-term storage using different diluents. Tested diluents included milk-based ones such as INRA 82 and Kenney’s diluent versus chemically defined diluent, INRA 96.
- The researchers used the milk fraction that demonstrated the best maintenance of sperm survival based on motility in previous studies.
- Four breeding trials were carried out to determine the influence of the new diluent and storage conditions on the fertility of the stallion.
Comparison of Protocols
- The standard protocol of storing semen involves using a skim milk diluent (INRA 82 or Kenney’s diluent) at 4 degrees Celsius under anaerobic conditions.
- Contrarily, the experimental protocol involves storing in a chemically defined, milk-free diluent (INRA 96), at 15 degrees Celsius, under aerobic conditions.
Observations and Findings
- Comparison of the two protocols showed a higher rate of fertility per cycle using the experimental protocol. After four breeding trials, fertility rates of the experimental and the standard protocol were 57% and 40% respectively.
- Further studies were conducted to see the effect of storage time on sperm fertility. An immediate insemination showed a 68% fertility per cycle, whereas semen stored under the experimental protocol for 72 hours before insemination showed a 48% fertility rate per cycle.
Conclusion
- The experimental protocol significantly improved sperm fertility compared to the standard protocol, presenting itself as a viable alternative especially for stallions with cold shock sensitive spermatozoa.
- This method could potentially be beneficial and applicable for semen storage in the field since storing semen for 72 hours under the experimental protocol still yielded reasonable fertility rates.
Cite This Article
APA
Batellier F, Duchamp G, Vidament M, Arnaud G, Palmer E, Magistrini M.
(2000).
Delayed insemination is successful with a new extender for storing fresh equine semen at 15 degrees C under aerobic conditions.
Theriogenology, 50(2), 229-236.
https://doi.org/10.1016/s0093-691x(98)00130-7 Publication
Researcher Affiliations
- Unité de reproduction équine, I.N.R.A.-Haras nationaux, PRMD, Nouzilly, France.
MeSH Terms
- Aerobiosis
- Animals
- Cold Temperature
- Female
- Horses
- Insemination, Artificial / veterinary
- Male
- Milk
- Pregnancy
- Pregnancy Rate
- Semen Preservation / methods
- Semen Preservation / veterinary
- Time Factors
Citations
This article has been cited 5 times.- Rečková Z, Filipčík R, Soušková K, Kopec T, Hošek M, Pešan V. The efficiency of different types of extenders for semen cooling in stallions. Anim Biosci 2022 May;35(5):670-676.
- Lindahl J, Dalin AM, Stuhtmann G, Morrell JM. Stallion spermatozoa selected by single layer centrifugation are capable of fertilization after storage for up to 96 h at 6°C prior to artificial insemination. Acta Vet Scand 2012 Jul 12;54(1):40.
- Morrell JM, Johannisson A, Juntilla L, Rytty K, Bäckgren L, Dalin AM, Rodriguez-Martinez H. Stallion Sperm Viability, as Measured by the Nucleocounter SP-100, Is Affected by Extender and Enhanced by Single Layer Centrifugation. Vet Med Int 2010;2010:659862.
- Becerro-Rey L, Martín-Cano FE, Silva-Rodríguez A, Ortega-Ferrusola C, da Silva-Álvarez E, Ortiz-Placín C, Tapia JA, Gil MC, Peña FJ. Stallion spermatozoa express LDH isoforms A, B, and C, with LDHC playing a crucial role in sustaining sperm viability. Reproduction 2025 Jul 1;170(1).
- Medica AJ, Gibb Z, Aitken RJ. Optimizing equine sperm quality: an alternative to single layer centrifugation for sperm isolation. Reprod Fertil 2024 Oct 1;5(4).
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