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Home / Equine Research Bank / Anim Reprod Sci / Advances in cooled semen technology.
Animal reproduction science2001; 68(3-4); 181-190; doi: 10.1016/s0378-4320(01)00155-5

Advances in cooled semen technology.

Abstract: In the horse industry, milk or milk-based extenders are used routinely for dilution and storage of semen cooled to 4-8 degrees C. Although artificial insemination (AI) with chilled and transported semen has been in use for several years, pregnancy rates are still low and variable related to variable semen quality of stallions. Over the years, a variety of extenders have been proposed for cooling, storage and transport of stallion semen. Fractionation of milk by microfiltration, ultrafiltration, diafiltration and freeze-drying techniques has allowed preparation of purified milk fractions in order to test them on stallion sperm survival. Finally, a high protective fraction, native phosphocaseinate (NPPC), was identified. A new extender, INRA96, based on modified Hanks' salts, supplemented with NPPC was then developed for use with cooled/stored semen. Four experiments were conducted to compare INRA96 and milk-based extenders under various conditions of storage. The diluted semen was maintained under aerobic conditions when stored at 15 degrees C, and anaerobic conditions when stored at 4 degrees C. In experiment 1, split ejaculates from 13 stallions were diluted either in INRA96 extender then stored at 15 degrees C or diluted in Kenney or INRA82 extenders and then stored at 4 degrees C for 24h, until insemination. In experiment 2, semen from two stallions was extended in INRA96 then inseminated immediately or stored at 15 degrees C for 3 days until insemination. In experiment 3, semen from three stallions was diluted in INRA96 then stored at 15 or 4 degrees C for 24h until insemination, finally, in experiment 4, split ejaculates from four stallions were diluted in INRA96 or E-Z Mixin extenders then stored at 4 degrees C for 24h until insemination. Experiment 1 demonstrated that at 15 degrees C, INRA96 extender significantly improved pregnancy rate per cycle compared to Kenney or INRA82 extenders at 4 degrees C after 24h of storage (57%, n=178 versus 40%, n=171, respectively; P<0.01). Experiment 2 showed that semen stored at 15 degrees C for 3 days can achieve pregnancy at a fertility rate per cycle of 48% (n=52) compared to 68% (n=50, immediate insemination, P=0.06). Experiment 3 demonstrated that INRA96 extender can be as efficient at 15 degrees C (54%, n=37) as at 4 degrees C (54%, n=35) after 24h of storage. Finally, experiment 4 showed that INRA96 extender used at 4 degrees C (59%, n=39) seems to improve fertility per cycle compared to E-Z Mixin at 4 degrees C (49%, n=39, P=0.25), but this result has to be confirmed. These results demonstrate that semen diluted in INRA96 extender and stored at 15 degrees C can be an alternative to semen diluted in milk-based extenders and stored at 4 degrees C for "poor cooler" stallions. Furthermore, INRA96 extender can be as efficient at 15 degrees C as at 4 degrees C, for preserving sperm motility and fertility.
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Publication Date: 2001-12-18 PubMed ID: 11744263DOI: 10.1016/s0378-4320(01)00155-5Google Scholar: Lookup
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  • Comparative Study
  • 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 focuses on the improvements made in the technology of cooling and storing horse semen for artificial insemination. The highlighted breakthrough is the introduction of a new extender, INRA96, which seems to improve the effectiveness in preserving sperm motility and fertility at both 15 and 4 degrees Celsius compared to traditional milk-based extenders.

Background and Goal of Study

  • In the horse industry, milk-based extenders are frequently used to dilute and store semen at 4-8 degrees Celsius. However, these extenders have been associated with low and inconsistent pregnancy rates.
  • The goal of the study was to investigate the effectiveness of a new extender dubbed ‘INRA96’ in improving the survival and motility of stallion sperm, consequently enhancing the resultant pregnancy rates in artificial insemination.

Methods Used and Findings

  • Four different experiments were conducted under varying conditions of storage for comparing INRA96 with existing milk-based extenders.
  • In experiment 1, split ejaculates from 13 stallions were diluted either in INRA96 extender, stored at 15 degrees Celsius, or in Kenney or INRA82 extenders and stored at 4 degrees Celsius for 24 hours. The result showed that INRA96 significantly improved pregnancy rate per cycle.
  • In experiment 2, semen from two stallions was extended in INRA96 then inseminated immediately or stored at 15 degrees Celsius for 3 days before insemination. The result indicated semen stored for 3 days can achieve pregnancy at a relatively comparable fertility rate to immediate insemination.
  • Experiment 3 involved semen from three stallions diluted in INRA96 then stored at 15 or 4 degrees Celsius for 24 hours. The result showed INRA96 can effectively maintain sperm fertility at either temperature.
  • In experiment 4, split ejaculates from four stallions were extended in INRA96 or E-Z Mixin extenders and stored at 4 degrees Celsius for 24 hours. The outcome suggested a higher fertility rate using the INRA96 extender compared to the E-Z Mixin, although this result still needs further confirmatory research.

Key Outcomes and further Research

  • The results demonstrated that the INRA96 extender used at 15 degrees Celsius can potentially replace milk-based extenders for stallions that fare poorly with cooler temperatures.
  • Additional research is needed to further confirm the observed improvement in fertility per cycle yield using INRA96 compared to other extenders, particularly for those stored at 4 degrees Celsius.

Cite This Article

APA
Batellier F, Vidament M, Fauquant J, Duchamp G, Arnaud G, Yvon JM, Magistrini M. (2001). Advances in cooled semen technology. Anim Reprod Sci, 68(3-4), 181-190. https://doi.org/10.1016/s0378-4320(01)00155-5

Publication

Animal reproduction science
ISSN: 0378-4320
NlmUniqueID: 7807205
Country: Netherlands
Language: English
Volume: 68
Issue: 3-4
Pages: 181-190

Researcher Affiliations

Batellier, F
  • Equipe Reproduction, INRA, Unité SRA, 37380 Nouzilly, France.
Vidament, M
    Fauquant, J
      Duchamp, G
        Arnaud, G
          Yvon, J M
            Magistrini, M

              MeSH Terms

              • Animals
              • Cryopreservation / methods
              • Cryopreservation / veterinary
              • Cryoprotective Agents / pharmacology
              • Female
              • Fertility
              • Horses / physiology
              • Insemination, Artificial / veterinary
              • Linear Models
              • Male
              • Milk / physiology
              • Pregnancy
              • Random Allocation
              • Semen
              • Semen Preservation / methods
              • Semen Preservation / veterinary
              • Sperm Motility

              Citations

              This article has been cited 18 times.
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