Effect of Caffeine and Pentoxifylline Added Before or After Cooling on Sperm Characteristics of Stallion Sperm.
Abstract: Different additives have been tested in cooled stallion sperm, in order to maintain sperm quality and to ameliorate the decrease in sperm fertility potential. In several species, caffeine and pentoxifylline promote sperm motility by increasing energy production. We evaluate the effects of caffeine and pentoxifylline when added to stallion sperm before or after cooling. Three ejaculates from five stallions each were processed and resuspended in skim milk extender. Caffeine (5 mM), pentoxifylline (3.5 mM), or both additives combined were included to sperm before or after cooling (4°C for 24 hours). Cooled sperm were incubated at 37°C and evaluated at 0, 30, 60, and 120 minutes for motility, morphology, viability (flow cytometry), and membrane functionality (hypo-osmotic swelling test). Results were analyzed by two-factor mixed model for repeated measures and Tukey comparisons. As main effects, the caffeine and pentoxifylline affected significantly motility and kinematic parameters, without interaction between treatment and incubation after cooling. No differences were observed whether the additives were added prior or after cooling. Pentoxifylline added after cooling reduced significantly motility during incubation, but with higher values at 30 minutes. We detected a decrease in morphologically normal sperm (P < .0001), caused by an increase of tail defects (P < .003) in the presence of both additives. Viability and membrane functionality were also significantly impaired by additives. Pentoxifylline when added after cooling improved sperm motility and kinematic parameters for a short period of time. However, sperm characteristic related to fertility potential was compromised after a prolonged exposure to caffeine or pentoxifylline.
Copyright © 2019 Elsevier Inc. All rights reserved.
Publication Date: 2019-12-26 PubMed ID: 32172905DOI: 10.1016/j.jevs.2019.102902Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study investigated the impact of caffeine and pentoxifylline on the quality of stallion sperm, when added before or after cooling. The experiments showed that both substances facilitated sperm motility, but also caused a decrease in normal sperm morphology and reduced sperm viability and membrane functionality.
Research Purpose
- The intention of this research was to explore the effects of two additives, caffeine and pentoxifylline, on the characteristics of cooled stallion sperm. The additives were tested to see if they could maintain the quality of sperm and potentially improve the fertility potential, which tends to decrease after cooling.
Methodology
- The research was completed using three ejaculates from five different stallions. The sperm were resuspended in a skim milk extender.
- Caffeine, pentoxifylline, or both were added to the sperm either before or after cooling them to 4°C for a 24-hour period.
- After cooling, the sperm were incubated at 37°C and evaluated at different time intervals (0, 30, 60, and 120 minutes) for motility, morphology, viability (measured by flow cytometry), and membrane functionality (checked through a hypo-osmotic swelling test).
Findings
- The research discovered that both caffeine and pentoxifylline significantly affected the sperm’s motility and kinematic parameters, regardless of whether they were added prior to or after cooling.
- Pentoxifylline, when added after cooling, was found to reduce sperm motility over time, although there was a brief period at 30 minutes post-cooling where motility showed higher values.
- The presence of both additives was associated with a decrease in the proportion of sperm with normal morphology, mainly due to an increase in tail defects.
- Sperm viability and membrane functionality were also seen to be significantly impaired by the addition of caffeine and pentoxifylline.
Conclusion
- Although pentoxifylline improved sperm motility and kinematic parameters when added after cooling, the study concluded that prolonged exposure to either caffeine or pentoxifylline appeared to compromise several key characteristics linked to the sperm’s fertility potential.
Cite This Article
APA
Rossi M, Gonzalez-Castro R, Falomo ME.
(2019).
Effect of Caffeine and Pentoxifylline Added Before or After Cooling on Sperm Characteristics of Stallion Sperm.
J Equine Vet Sci, 87, 102902.
https://doi.org/10.1016/j.jevs.2019.102902 Publication
Researcher Affiliations
- Department of Animal Medicine, Production and Health (MAPS), Università di Padova, Legnaro (PD), Italy. Electronic address: melissa.rossi@unipd.it.
- Equine Reproduction Laboratory, Colorado State University, Fort Collins, CO, USA.
- Department of Animal Medicine, Production and Health (MAPS), Università di Padova, Legnaro (PD), Italy.
MeSH Terms
- Animals
- Caffeine / pharmacology
- Fertility
- Horses
- Male
- Pentoxifylline / pharmacology
- Sperm Motility
- Spermatozoa
Citations
This article has been cited 4 times.- Boni R, Ruggiero R, De Luca F, Preziosi G, Ferrara MA, Ostuni A, Guerriero S, Gallo A, Murano C, Cecchini Gualandi S. Enhancing Stallion Semen Cryopreservation: Selected Antioxidant Extracts and Sperm Freezability. Antioxidants (Basel) 2025 Nov 16;14(11).
- Aliabadi E, Nateghian Z, Nasr-Esfahani MH, Tavalaee M, Talaei-Khozani T. Effects of L-carnitine and pentoxifylline on long-term preservation of the human sperms: An experimental study. Int J Reprod Biomed 2024 Nov;22(11):871-882.
- Raoofi A, Gholami O, Mokhtari H, Bagheri F, Rustamzadeh A, Nasiry D, Ghaemi A. Caffeine attenuates spermatogenic disorders in mice with induced chronic scrotal hyperthermia. Clin Exp Reprod Med 2024 Mar;51(1):28-41.
- Sun SY, Li Y, Gao YY, Ran XW. Efficacy and Safety of Pentoxifylline for Venous Leg Ulcers: An Updated Meta-Analysis. Int J Low Extrem Wounds 2024 Jun;23(2):264-274.
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