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Theriogenology2020; 157; 85-95; doi: 10.1016/j.theriogenology.2020.07.027

Irradiating frozen-thawed stallion sperm with red-light increases their resilience to withstand post-thaw incubation at 38 °C.

Abstract: The aim of this study was to evaluate whether red-light stimulation increases the longevity and resilience of cryopreserved stallion sperm to withstand post-thaw incubation for 120 min. Sixteen frozen straws of 0.5 mL from eight stallions were used. Samples were cryopreserved, thawed through incubation at 38 °C for 30 s and divided into the control and samples exposed to red-light using a triple LED photo-activation system (wavelength: 620-630 nm). Three irradiation protocols consisting of different light-dark-light intervals (1-1-1, 2-2-2 and 3-3-3 min) were tested. Sperm quality parameters were analyzed immediately after light-stimulation (0 min) and after 120 min of incubation at 38 °C. Sperm motility was evaluated using a Computerized Semen Analysis System (CASA), and flow cytometry and different fluorochromes were used to evaluate the integrity and lipid disorder of plasma membrane, mitochondrial membrane potential and intracellular levels of peroxides and superoxides. Irradiation significantly increased the percentages of spermatozoa with high mitochondrial membrane potential (1-1-1 pattern) and the intracellular levels of peroxides (2-2-2 pattern) at 0 min. In addition, sperm kinematic parameters (2-2-2 and 3-3-3 patterns) and percentages of viable spermatozoa with low membrane lipid disorder (3-3-3 pattern) were significantly higher in irradiated samples than in the control at 120 min. Our results indicate that red-light stimulation could help increase the resilience of frozen-thawed stallion sperm to withstand post-thaw incubation at 38 °C for 120 min and that these effects rely on the irradiation pattern. Further research should evaluate whether light-stimulation could also have a positive on fertility rates after artificial insemination.
Copyright © 2020 Elsevier Inc. All rights reserved.
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Publication Date: 2020-07-28 PubMed ID: 32805646DOI: 10.1016/j.theriogenology.2020.07.027Google 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.

This research investigates if exposing cryopreserved (frozen, then thawed) stallion sperm to red light can heighten their survivability, particularly by enhancing their ability to endure a 120-minute post-thaw incubation at 38 degrees Celsius.

Study Methodology

  • The research utilized frozen sperm samples from eight stallions, with a total of 16 0.5 mL straws.
  • The samples underwent cryopreservation, were thawed via incubation at 38 degrees Celsius for 30 seconds, then segregated into a control group and red-light exposed samples.
  • The research employed a triple LED photo-activation system emitting red light, with a wavelength of 620-630 nm.
  • The irradiation protocols consisted of three distinct light-dark-light intervals lasting 1-1-1, 2-2-2, and 3-3-3 minutes, respectively.
  • Sperm quality parameters were analyzed immediately after light-stimulation and after incubation at 38 degrees Celsius for 120 minutes.

Sperm Evaluation Techniques

  • Sperm motility was assessed using a Computerized Semen Analysis System (CASA).
  • Flow cytometry and different fluorochromes were used to evaluate the integrity and lipid disorder of the plasma membrane, mitochondrial membrane potential, and intracellular levels of peroxides and superoxides.

Research Findings

  • Significant increases were observed in the percentages of spermatozoa with high mitochondrial membrane potential (1-1-1 pattern) and intracellular levels of peroxides (2-2-2 pattern) at 0 minutes post-irradiation.
  • After 120 minutes, sperm kinematic parameters (2-2-2 and 3-3-3 patterns), and the percentages of viable spermatozoa with low membrane lipid disorder (3-3-3 pattern) were proven to be significantly higher in irradiated samples than in the control group.
  • Suggesting that red-light stimulation could potentially enhance the survivability of frozen-thawed stallion sperm post-thaw incubation at 38 degrees Celsius for 120 minutes, with effects varying based on the irradiation pattern used.

Future Research

  • As an extension of this study, the researchers propose exploring whether light-stimulation could also positively impact fertility rates after artificial insemination.

Cite This Article

APA
Catalán J, Llavanera M, Bonilla-Correal S, Papas M, Gacem S, Rodríguez-Gil JE, Yeste M, Miró J. (2020). Irradiating frozen-thawed stallion sperm with red-light increases their resilience to withstand post-thaw incubation at 38 °C. Theriogenology, 157, 85-95. https://doi.org/10.1016/j.theriogenology.2020.07.027

Publication

Theriogenology
ISSN: 1879-3231
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 157
Pages: 85-95

Researcher Affiliations

Catalán, Jaime
  • Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain; Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, Girona, Spain; Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, Girona, Spain.
Llavanera, Marc
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, Girona, Spain; Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, Girona, Spain.
Bonilla-Correal, Sebastián
  • Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain; Faculty of Veterinary Medicine, Antonio Nariño University, Bogotá, Colombia.
Papas, Marion
  • Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain.
Gacem, Sabrina
  • Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain.
Rodríguez-Gil, Joan E
  • Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain.
Yeste, Marc
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, Girona, Spain; Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, Girona, Spain. Electronic address: marc.yeste@udg.edu.
Miró, Jordi
  • Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain.

MeSH Terms

  • Animals
  • Cryopreservation / veterinary
  • Freezing
  • Horses
  • Male
  • Semen Preservation / veterinary
  • Sperm Motility
  • Spermatozoa

Conflict of Interest Statement

Declaration of competing interest J.E.R.G. and M.Y. are inventors of a patent entitled ‘Method and apparatus for improving the quality of mammalian sperm’ (European Patent Office, No. 16199093.2; EP-3-323-289-A1), which is owned by Instruments Utils de Laboratori Geniul, SL (Barcelona, Spain).

Citations

This article has been cited 9 times.
  1. Blanco-Prieto O, Maside C, Gruzmacher A, Ortiz M, Ratto M, Urra FJ, Vera T, Strobel P, Catalán J, Mislei B, Bucci D, Yeste M, Rodríguez-Gil JE, Ramírez-Reveco A. Red LED Light Irradiation Increases the Resistance Against Environmental Stress of Frozen Bovine Sperm Thawed in Suboptimal Conditions. Animals (Basel) 2025 Nov 20;15(22).
    doi: 10.3390/ani15223353pubmed: 41302061google scholar: lookup
  2. Ullah A, Chen W, Shi L, Wang M, Geng M, Na J, Akhtar MF, Khan MZ, Wang C. Challenges and Enhancing Strategies of Equine Semen Preservation: Nutritional and Genetic Perspectives. Vet Sci 2025 Aug 25;12(9).
    doi: 10.3390/vetsci12090807pubmed: 41012733google scholar: lookup
  3. Castro M, Leal K, Pezo F, Contreras MJ. Sperm Membrane: Molecular Implications and Strategies for Cryopreservation in Productive Species. Animals (Basel) 2025 Jun 19;15(12).
    doi: 10.3390/ani15121808pubmed: 40564358google scholar: lookup
  4. Moradi A, Ghaffari Novin M, Bayat M. A Comprehensive Systematic Review of the Effects of Photobiomodulation Therapy in Different Light Wavelength Ranges (Blue, Green, Red, and Near-Infrared) on Sperm Cell Characteristics in Vitro and in Vivo. Reprod Sci 2024 Nov;31(11):3275-3302.
    doi: 10.1007/s43032-024-01657-xpubmed: 39095677google scholar: lookup
  5. Dordas-Perpinyà M, Yánez-Ortiz I, Sergeant N, Mevel V, Catalán J, Bruyas JF, Miró J, Briand-Amirat L. ProAKAP4 as Indicator of Long-Lasting Motility Marker in Post-Thaw Conditions in Stallions. Animals (Basel) 2024 Apr 23;14(9).
    doi: 10.3390/ani14091264pubmed: 38731267google scholar: lookup
  6. Blanco-Prieto O, Maside C, Àlex Peña, Ibáñez-Príncep J, Bonet S, Yeste M, Rodríguez-Gil JE. The effects of red LED light on pig sperm function rely upon mitochondrial electron chain activity rather than on a PKC-mediated mechanism. Front Cell Dev Biol 2022;10:930855.
    doi: 10.3389/fcell.2022.930855pubmed: 36274839google scholar: lookup
  7. Catalán J, Yánez-Ortiz I, Tvarijonaviciute A, González-Aróstegui LG, Rubio CP, Barranco I, Yeste M, Miró J. Seminal Plasma Antioxidants Are Related to Sperm Cryotolerance in the Horse. Antioxidants (Basel) 2022 Jun 28;11(7).
    doi: 10.3390/antiox11071279pubmed: 35883774google scholar: lookup
  8. Catalán J, Yánez-Ortiz I, Gacem S, Papas M, Bonet S, Rodríguez-Gil JE, Yeste M, Miró J. The Effects of Red Light on Mammalian Sperm Rely upon the Color of the Straw and the Medium Used. Animals (Basel) 2021 Jan 8;11(1).
    doi: 10.3390/ani11010122pubmed: 33429933google scholar: lookup
  9. Catalán J, Papas M, Trujillo-Rojas L, Blanco-Prieto O, Bonilla-Correal S, Rodríguez-Gil JE, Miró J, Yeste M. Red LED Light Acts on the Mitochondrial Electron Chain of Donkey Sperm and Its Effects Depend on the Time of Exposure to Light. Front Cell Dev Biol 2020;8:588621.
    doi: 10.3389/fcell.2020.588621pubmed: 33365309google scholar: lookup
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