Comparison of the Effects of Five Semen Extenders on the Quality of Frozen-Thawed Equine Epididymal Sperm.
Abstract: Cryopreservation of epididymal sperm allows the saving of genetic material in case of unexpected death or emergency castration. The aim of the present study was the comparison of five different combinations of extenders commercially available for equine frozen semen processing for cryopreservation of epididymal sperm. Epididymal sperm were harvested from gonads of 10 healthy stallions after routine castration by retrograde flush technique. Then, samples were split and diluted with (1) INRA96 + INRA Freeze, (2) BotuSemen + BotuCRIO, (3) EquiPlus + Gent Freeze, (4) EquiPlus + EquiPlus Freeze, and (5) Gent + Gent Freeze. Extenders 1 and 2 showed higher values for total and progressive motility after thawing compared with extender 4 (P .05), and extender 5 resulted in the lowest values (P < .05). The subpopulation of viable frozen-thawed sperm with high mitochondrial membrane potential and low intracellular calcium content was higher using extender 1 compared with extenders 3, 4, and 5 (P < .05) and higher in extender 2 compared with extenders 4 and 5 immediately after thawing (P < .05). After 1 hour of incubation, this subpopulation yielded the highest values in extender 2 (P < .05). Immediately after thawing, extender 1 yielded higher values for percentage of DFI and mean DFI than extenders 3, 4, and 5 (P < .05). Following 1 hour of incubation after thawing, sperm processed with extender 1 resulted in the highest values for percentage of DFI and mean DFI (P < .05). Using extender 2, mean DFI values were lower than those in extender 1 and higher than the extenders 3, 4, and 5 (P < .05). The study revealed that according to the examined sperm quality parameters, freezing extenders (extender 1, extender 2) using low concentrations of glycerol either combined with or without methylformamide were beneficial for cryopreservation of stallion epididymal sperm. For processing of stallion epididymal sperm, an extender containing milk proteins (extenders 1-4) for initial dilution after sperm harvesting is preferable to an extender including egg yolk (extender 5).
Copyright © 2019 Elsevier Inc. All rights reserved.
Publication Date: 2019-05-16 PubMed ID: 31405486DOI: 10.1016/j.jevs.2019.05.002Google Scholar: Lookup
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Summary
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The research article discusses the investigation of five commercially available mixtures of extenders for freezing horse sperm in order to determine which one maintains the highest quality after thawing. The study found that the extenders containing lower concentrations of glycerol, either with or without methylformamide, performed best in preserving the quality of the horse’s sperm compared to the others.
Objective of the Study
- The objective of the study was to compare five different extenders used in the process of freezing equine (horse) sperm. The extenders which contained INRA96 + INRA Freeze, BotuSemen + BotuCRIO, EquiPlus + Gent Freeze, EquiPlus + EquiPlus Freeze, and Gent + Gent Freeze were evaluated for their effectiveness.
Methodology
- The research utilised sperm that was harvested from the epididymis of 10 healthy stallions after routine castration. The harvesting was performed through the retrograde flush technique.
- These samples were divided and diluted using the five different extenders under investigation.
Results
- The results showed that the extenders INRA96 + INRA Freeze and BotuSemen + BotuCRIO demonstrated superior values for total and progressive motility after thawing compared to EquiPlus + EquiPlus Freeze.
- EquiPlus + Gent Freeze showed results in between those of INRA96 + INRA Freeze and BotuSemen + BotuCRIO, with Gent + Gent Freeze providing the lowest values.
- In regards to cellular health of the sperm, the study found the highest amount of viable frozen-thawed sperm with good mitochondrial health and low calcium content in the samples prolonged with INRA96 + INRA Freeze and BotuSemen + BotuCRIO.
Conclusion
- Overall, the research showed the extenders INRA96 + INRA Freeze and BotuSemen + BotuCRIO, which use low concentrations of glycerol in combination with or without methylformamide, were the most effective at preserving the quality of frozen horse sperm.
- The study also found that extenders containing milk proteins (Extenders 1-4) are better for initial dilution after sperm harvesting as compared to an extender containing egg yolk (Extender 5).
Cite This Article
APA
Neuhauser S, Bollwein H, Siuda M, Handler J.
(2019).
Comparison of the Effects of Five Semen Extenders on the Quality of Frozen-Thawed Equine Epididymal Sperm.
J Equine Vet Sci, 79, 1-8.
https://doi.org/10.1016/j.jevs.2019.05.002 Publication
Researcher Affiliations
- Pferdezentrum Bad Saarow, Equine Reproduction Unit, Freie Universität Berlin, Bad Saarow, Germany.
- Clinic of Reproductive Medicine, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland.
- Clinic of Reproductive Medicine, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland.
- Pferdezentrum Bad Saarow, Equine Reproduction Unit, Freie Universität Berlin, Bad Saarow, Germany. Electronic address: johannes.handler@fu-berlin.de.
MeSH Terms
- Animals
- Cryopreservation / veterinary
- Cryoprotective Agents
- Freezing
- Horses
- Humans
- Male
- Semen
- Sperm Motility
- Spermatozoa
Citations
This article has been cited 3 times.- Al-Khaldi K, Yimer N, Sadiq MB, Firdaus Jesse Bin Abdullah F, Salam Babji A, Al-Bulushi S. Edible bird's nest supplementation in chilled and cryopreserved Arabian stallion semen.. Saudi J Biol Sci 2022 Mar;29(3):1577-1584.
- Al-Khaldi K, Yimer N, Al-Bulushi S, Haron AW, Hiew M, Babji AS. A preliminary study on the effects of E-Z Mixin® and EquiPlus® extenders supplemented with Edible Bird's Nest on the quality of chilled Arabian stallion semen.. Anim Reprod 2021 Jun 21;18(2):e20200027.
- Bustani GS, Baiee FH. Semen extenders: An evaluative overview of preservative mechanisms of semen and semen extenders.. Vet World 2021 May;14(5):1220-1233.
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