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Effects of glucose concentration in semen extender and storage temperature on stallion sperm quality following long-term cooled storage.
Abstract: In Experiment 1, the effects of glucose concentration in extender (0 mM, 67 mM, 147 mM, 270 mM; G0, G67, G147, and G270, respectively) and storage temperature of extended semen (5, 10, 15 and 20 °C) were evaluated after storage for up to 5 days (T0h to T120h). For all time points tested, mean total (TMOT) and progressive (PMOT) sperm motility were lower in G0 than all other treatment groups (P < 0.05). Mean curvilinear velocity (VCL) was lower in G0 than other treatment groups at all time points tested except T0h (P 0.05). Mean TMOT and PMOT, were lower for semen stored at 20 °C than all lower storage temperatures (P < 0.05) at all time points. In Experiment 2, semen was stored at 10 °C in extender containing no added glucose (G0) or 147 mM glucose (G147). Following storage, semen was centrifuged and resuspended in extender containing no added glucose (G0 - G0 or G147 - G0, respectively) or 147 mM of glucose (G0 - G147 or G147 - G147, respectively). Mean TMOT, PMOT, and VCL were higher in G147 than G0 at all time periods tested (P 0.05). Mean TMOT and PMOT were higher in G0 - G147 than G0 - G0 at T72h and T120h (P < 0.05) and mean VCL was higher in G0 - G147 than G0 - G0 for all time periods. Mean TMOT, PMOT, and VCL were higher in G147 - G147 than G147 - G0 at all time points tested (P 0.05). In Experiment 3, the minimum concentration of glucose required to maintain sperm quality following long-term cooled storage (T120 h) was evaluated (G0, G5, G10, G20, G40, G67, G147 mM). At T120 h, mean TMOT was lowest in G0, G5, G10, and G20 (P < 0.05), whereas mean PMOT and VCL were lower in G0, G5, G10, and G20 than in G40, G67, and G147 (P 0.05). In conclusion, the absence of added glucose in extender reduced the motion characteristics of stallion sperm during long-term storage (5 days), but VAI was not affected. The use of temperatures between 5 and 15 °C for long-term storage (5 days) best maintained sperm motility and VAI. The threshold concentration of added glucose in extender required to optimize sperm motion characteristics was 40 mM.
Published by Elsevier Inc.
Publication Date: 2020-02-11 PubMed ID: 32070880DOI: 10.1016/j.theriogenology.2020.02.007Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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This research demonstrates that the concentration of glucose in the extender used for preserving stallion sperm and the temperature at which it’s stored significantly affects the sperm’s quality over long-term cool storage, with optimal conditions identified for best preserving sperm motility.
Research Methodology
The research was conducted in three experiments.
- In the first experiment, the effects of glucose concentrations (0 mM, 67 mM, 147 mM, and 270 mM) and storage temperatures (5°C, 10°C, 15°C, and 20°C) were assessed over a span of five days. Sperm quality was evaluated throughout, based on mean total (TMOT) and progressive (PMOT) sperm motility, curvilinear velocity (VCL), and percentage of plasma/acrosome intact sperm (VAI).
- In the second experiment, the researchers stored semen at 10°C both with and without added glucose. The stored semen was then centrifuged and resuspended in extenders both with and without added glucose to explore differences in sperm quality.
- In the third experiment, the minimum glucose concentration needed to maintain sperm quality for long-term cooled storage was evaluated. Glucose concentrations from 0 mM up to 147 mM were used and sperm quality was evaluated after 120 hours.
Findings
In the first experiment, the results showed that mean total and progressive sperm motility were lowest in the absence of glucose (0 mM) at all time points, while mean curvilinear velocity was lower at all time points except at the start of the experiment. The percentage of plasma/acrosome intact sperm was similar across all treatments at the start, midpoint, and endpoint (120 hours). The research also showed that sperm stored at 20°C had reduced overall and progressive motility compared to lower storage temperatures.
In the second experiment, total and progressive motility as well as VCL were found to be higher with added glucose (147 mM) in extenders during storage and resuspension processes. The percentage of plasma/acrosome intact sperm remained unaffected by whether glucose was added or not.
Third experiment suggested the threshold concentration of added glucose in the extender required to optimize sperm motion characteristics was 40 mM.
Conclusion
The researchers concluded that the lack of added glucose in the extender reduced the motion characteristics of stallion sperm during long-term 5-day storage, though it had no impact on the percentage of plasma/acrosome intact sperm. The most effective temperature for long-term storage was found to be between 5 and 15°C.
Cite This Article
APA
Hernández-Avilés C, Love CC, Serafini R, Ramírez-Agámez L, Friedrich M, Ghosh S, Teague SR, LaCaze KA, Brinsko SP, Varner DD.
(2020).
Effects of glucose concentration in semen extender and storage temperature on stallion sperm quality following long-term cooled storage.
Theriogenology, 147, 1-9.
https://doi.org/10.1016/j.theriogenology.2020.02.007 Publication
Researcher Affiliations
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, USA. Electronic address: chernandez@cvm.tamu.edu.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, USA.
MeSH Terms
- Animals
- Cell Survival / drug effects
- Cryoprotective Agents / pharmacology
- Glucose / pharmacology
- Horses
- Male
- Semen
- Semen Analysis / veterinary
- Semen Preservation / methods
- Semen Preservation / veterinary
- Sperm Motility
- Spermatozoa / drug effects
- Temperature
Conflict of Interest Statement
Declarations of competing interest None.
Citations
This article has been cited 8 times.- Allai L, Druart X, Terzioğlu P, Louanjli N, Nasser B, Ozturk M, El Amiri B. The Addition of Opuntia ficus-indica Ethanolic Extract to a Skimmed Milk-Based Extender Impacts Ram Sperm Quality. Vet Med Int 2023;2023:6248890.
- Cheng Q, Li L, Jiang M, Liu B, Xian Y, Liu S, Liu X, Zhao W, Li F. Extend the Survival of Human Sperm In Vitro in Non-Freezing Conditions: Damage Mechanisms, Preservation Technologies, and Clinical Applications. Cells 2022 Sep 12;11(18).
- 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.
- 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.
- Alam ME, Yeasmin MS, Das DK, Islam MS, Ahmed S, Islam MH, Islam MA, Kamal MM, Kabir A, Khan A, Haque MH, Masum MA, Rahman M, Moni MIZ. Comparative study of the glucose and trehalose addition on the extenders for goat sperm liquid storage. J Adv Vet Anim Res 2025 Mar;12(1):287-296.
- Arif A, Zahoor N, Tang J, Tang M, Dong L, Khan SZ, Dai G. Cryopreservation Strategies for Poultry Semen: A Comprehensive Review of Techniques and Applications. Vet Sci 2025 Feb 8;12(2).
- Al-Kass Z, Morrell JM. Freezing Stallion Semen-What Do We Need to Focus on for the Future?. Vet Sci 2024 Feb 2;11(2).
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