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Home / Equine Research Bank / Animals (Basel) / Retrograde Flushing Followed by Slicing Float-Up as an Appro...
Animals : an open access journal from MDPI2022; 12(14); 1802; doi: 10.3390/ani12141802

Retrograde Flushing Followed by Slicing Float-Up as an Approach to Optimize Epididymal Sperm Recovery for the Purpose of Cryopreservation in Equids.

Abstract: This study aimed to assess the parameters of epididymal sperm harvested by retrograde flushing (RF) followed by slicing float-up (SF). Epididymides from donkeys (n = 18) and horses (n = 28) were subjected to RF with a freezing extender and then SF technique. The retrieved sperm after RF and SF was evaluated for volume, concentration, and total sperm and then cryopreserved separately. Post-thaw total motility (TM) and progressive motility (PM) were evaluated with CASA. Sperm membrane integrity (SMI) and mitochondrial membrane potential (MMP) were assessed with flow cytometry. Sperm concentration was greater in donkeys than horses (684 ± 62.9 vs. 494 ± 50.9 million sperm/mL) (p = 0.02). The total sperm harvested was lower in SF (3.6 ± 0.7 billion) than RF (10.4 ± 1.5 billion) and in horses (4.6 ± 0.8 billion) than in donkeys (10.7 ± 1.8 billion) (p 0.05). Post-thawing SMI and MMP did not vary with technique or species (p > 0.05); TM and PM were not influenced by the technique or the species (p > 0.05) but by their interaction (p = 0.005). In conclusion, using RF followed by SF enhances sperm recovery without affecting cryopreservation in equids.
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Publication Date: 2022-07-14 PubMed ID: 35883349PubMed Central: PMC9312152DOI: 10.3390/ani12141802Google 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.

The research paper evaluates the optimum procedure of epididymal sperm recovery in equines for cryopreservation, by conducting two techniques— Retrograde Flushing (RF) and Slicing Float-up (SF) on horses and donkeys. The researchers measured the sperm parameters and found that using both methods together significantly increased the amount of sperm recovered without impacting the process of freezing and thawing the sperm.

Methodology and Results

  • The study involved two species of equids: donkeys (18 in number) and horses (28 in number).
  • The researchers collected the epididymides (a part of the male reproductive system where sperm is stored) from these animals. They first used the RF technique, in which a freezing extender is applied, followed by the SF technique.
  • They then measured the volume, concentration, and total number of sperm collected using the two methods.
  • The researchers found that the concentration of sperm was higher in donkeys than in horses, with an average of 684 million sperm/mL compared to 494 million sperm/mL.
  • However, the total sperm harvested was lower with the SF method (3.6 billion) compared to the RF method (10.4 billion). It was also lower in horses (4.6 billion) compared to donkeys (10.7 billion).
  • The use of both RF and SF resulted in a 57% and 31% increase in sperm harvested from donkeys and horses, respectively.

Sperm Quality Assessment

  • Total Motility (TM) and Progressive Motility (PM), which are parameters used to measure the movement and direction of sperm, were left unaffected by the technique used or the species of the animal.
  • The researchers also tested Sperm Membrane Integrity (SMI) and Mitochondrial Membrane Potential (MMP) using flow cytometry. These are tests that assess the health and functionality of sperm cells.
  • These three parameters— SMI, MMP, and TM did not differ significantly between the RF and SF techniques or between the species.

Conclusion

  • The study concludes that employing both retrograde flushing and slicing float-up techniques together helps to yield a higher amount of sperm.
  • The use of both methods does not affect the freezing and thawing processes which are essential in cryopreservation of sperm, thereby making this a useful approach in sperm recovery in equids.

Cite This Article

APA
Podico G, Canisso IF. (2022). Retrograde Flushing Followed by Slicing Float-Up as an Approach to Optimize Epididymal Sperm Recovery for the Purpose of Cryopreservation in Equids. Animals (Basel), 12(14), 1802. https://doi.org/10.3390/ani12141802

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 12
Issue: 14
PII: 1802

Researcher Affiliations

Podico, Giorgia
  • Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA.
Canisso, Igor F
  • Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA.

Conflict of Interest Statement

The authors declare no conflict of interest.

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Citations

This article has been cited 4 times.
  1. Huijsmans TERG, Hassan HA, Smits K, Van Soom A. Postmortem Collection of Gametes for the Conservation of Endangered Mammals: A Review of the Current State-of-the-Art. Animals (Basel) 2023 Apr 15;13(8).
    doi: 10.3390/ani13081360pubmed: 37106923google scholar: lookup
  2. Podico G, Spencer KM, Magalhaes HB, Canisso IF. Semen Quality of the First and Second Ejaculates Collected from Breeding Inactive Stallions after Cooling and Freezing. Vet Sci 2023 Feb 21;10(3).
    doi: 10.3390/vetsci10030173pubmed: 36977212google scholar: lookup
  3. Bernklau E, Wehrend A, Farshad A. Evaluation of Sperm Retrieval Efficiency and Extender Impact in Cryopreserved Canine Epididymal Semen. Vet Sci 2025 Aug 31;12(9).
    doi: 10.3390/vetsci12090840pubmed: 41012765google scholar: lookup
  4. Santiani A, Cucho M, Delgado J, Juárez J, Ruiz L, Evangelista-Vargas S. Field-Compatible Cytometric Assessment of Epididymal Alpaca Sperm Viability and Acrosomal Integrity Using Fluorochrome. Animals (Basel) 2025 Aug 5;15(15).
    doi: 10.3390/ani15152282pubmed: 40805072google scholar: lookup
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