Liquid storage of equine semen: Assessing the effect of d-penicillamine on longevity of ejaculated and epididymal stallion sperm.
Abstract: Short-term storage of equine sperm at 5°C in an extender containing milk and/or egg yolk components is common practice in the equine breeding industry. Sperm motility, viability, DNA integrity and, consequently, fertilizing ability decline over time, partly due to reactive oxygen species (ROS) generation. We investigated whether adding the anti-oxidant d-penicillamine to a commercial milk/egg yolk extender delayed the decrease in semen quality. Semen was recovered on four consecutive days from eight 3-year old Warmblood stallions. On day 5, seven of the stallions were castrated and sperm recovered from the caudae epididymides. Ejaculated samples were split, and one portion was centrifuged and re-suspended to reduce seminal plasma content. All samples were diluted to 50millionsperm/ml and divided into two portions, one of which was supplemented with 0.5mM d-penicillamine. After 48h, 96h, 144h and 192h storage, sperm motility was assessed by computer-assisted semen analysis (CASA), viability by SYBR14/PI staining, and DNA integrity using the sperm chromatin structure assay (SCSA). d-Penicillamine had no effect on motility of ejaculated sperm (P>0.05) but reduced total and progressive motility of epididymal sperm. Sperm chromatin integrity was not influenced by storage time, seminal plasma or d-penicillamine. In short, adding d-penicillamine to a commercial semen extender was neither beneficial nor detrimental to the maintenance of quality in ejaculated semen stored at 5°C. The negative effect on motility of epididymal sperm may reflect differences in (membrane) physiology of spermatozoa that have not been exposed to seminal plasma.
Copyright © 2015. Published by Elsevier B.V.
Publication Date: 2015-06-19 PubMed ID: 26130601DOI: 10.1016/j.anireprosci.2015.06.011Google Scholar: Lookup
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- Journal Article
Summary
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The research investigated if the antioxidant d-penicillamine could enhance the quality of stored stallion sperm. However, results show that the inclusion of the antioxidant neither improved nor harmed the quality of the ejaculated semen stored at 5°C, and negatively affected the mobility of the sperm retrieved from the epididymis.
Research Method
- Semen was collected over four days from eight Warmblood stallions, aged three years.
- On the fifth day, seven of the horses were castrated, and sperm was gathered from the caudae epididymides.
- Each ejaculated sample was divided. One portion was centrifuged and suspended again to decrease the seminal plasma content.
- All samples were diluted to 50 million sperm per milliliter before being divided into two, one portion of which was supplemented with 0.5mM d-penicillamine, an antioxidant.
- The quality of sperm was evaluated after 48, 96, 144, and 192 hours of storage.
- Computer-assisted semen analysis (CASA) determined sperm motility, while SYBR14/PI staining assessed viability, and DNA integrity was analyzed using the sperm chromatin structure assay (SCSA).
Findings
- d-Penicillamine did not significantly affect the mobility of the ejaculated sperm.
- The antioxidant reduced total and progressive motility of epididymal sperm, indicating a negative effect.
- Sperm chromatin integrity was not influenced by the storage time, level of seminal plasma, or d-penicillamine.
Conclusion
- Adding d-penicillamine to a commercial semen extender neither benefitted nor harmed the quality of ejaculated semen stored at 5°C.
- The negative effect seen on epididymal sperm movement may point to differences in the physiology of sperm cells that haven’t been exposed to seminal plasma.
Implication
- This study suggests that the addition of d-penicillamine in storage is not beneficial for preserving the quality of ejaculated stallion sperm.
- More research is needed to understand the different characteristics and behaviors of ejaculated and epididymal sperm in horses, which could help develop more effective preservation methods.
Cite This Article
APA
Brogan PT, Beitsma M, Henning H, Gadella BM, Stout TA.
(2015).
Liquid storage of equine semen: Assessing the effect of d-penicillamine on longevity of ejaculated and epididymal stallion sperm.
Anim Reprod Sci, 159, 155-162.
https://doi.org/10.1016/j.anireprosci.2015.06.011 Publication
Researcher Affiliations
- Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 114, 3584 CM Utrecht, The Netherlands. Electronic address: P.T.Brogan@uu.nl.
- Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 114, 3584 CM Utrecht, The Netherlands. Electronic address: M.M.Beitsma@uu.nl.
- Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 114, 3584 CM Utrecht, The Netherlands. Electronic address: H.H.W.Henning@uu.nl.
- Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 7, 3584 CM Utrecht, The Netherlands; Department of Biochemistry and Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 2, 3584 CM Utrecht, The Netherlands. Electronic address: B.M.Gadella@uu.nl.
- Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 114, 3584 CM Utrecht, The Netherlands; Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 7, 3584 CM Utrecht, The Netherlands; Section of Reproduction, Faculty of Veterinary Science, University of Pretoria, Private Bag XO4, Onderstepoort 0110, South Africa. Electronic address: T.A.E.Stout@uu.nl.
MeSH Terms
- Animals
- Ejaculation
- Epididymis
- Horses / physiology
- Male
- Penicillamine / pharmacology
- Semen / drug effects
- Semen Preservation / methods
- Semen Preservation / veterinary
Citations
This article has been cited 6 times.- Tourmente M, Sanchez-Rodriguez A, Roldan ERS. Effect of Motility Factors D-Penicillamine, Hypotaurine and Epinephrine on the Performance of Spermatozoa from Five Hamster Species.. Biology (Basel) 2022 Mar 30;11(4).
- Leemans B, Bromfield EG, Stout TAE, Vos M, Van Der Ham H, Van Beek R, Van Soom A, Gadella BM, Henning H. Developing a reproducible protocol for culturing functional confluent monolayers of differentiated equine oviduct epithelial cells†.. Biol Reprod 2022 Apr 26;106(4):710-729.
- Maitan PP, Bromfield EG, Hoogendijk R, Leung MR, Zeev-Ben-Mordehai T, van de Lest CH, Jansen JWA, Leemans B, Guimarães JD, Stout TAE, Gadella BM, Henning H. Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa.. Front Cell Dev Biol 2021;9:772254.
- Umair M, Henning H, Stout TAE, Claes A. A Modified Flotation Density Gradient Centrifugation Technique Improves the Semen Quality of Stallions with a High DNA Fragmentation Index.. Animals (Basel) 2021 Jul 1;11(7).
- Liu T, Han Y, Zhou T, Zhang R, Chen H, Chen S, Zhao H. Mechanisms of ROS-induced mitochondria-dependent apoptosis underlying liquid storage of goat spermatozoa.. Aging (Albany NY) 2019 Sep 23;11(18):7880-7898.
- Berlinguer F, Pasciu V, Succu S, Cossu I, Caggiu S, Addis D, Castagna A, Fontani V, Rinaldi S, Passino ES. REAC technology as optimizer of stallion spermatozoa liquid storage.. Reprod Biol Endocrinol 2017 Feb 8;15(1):11.
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