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Animal reproduction science2007; 105(1-2); 119-128; doi: 10.1016/j.anireprosci.2007.11.010

Commercial semen freezing: individual male variation in cryosurvival and the response of stallion sperm to customized freezing protocols.

Abstract: One of the challenges for those attempting to cryopreserve stallion spermatozoa is dealing with the stallion to stallion variability in the cryosurvival of their semen. In the dairy industry, each bull stud, essentially utilizes a single cryopreservation technique, and bulls that produce sperm that do not cryopreserve well using that technique are replaced by other bulls. However, replacing stallions is unlikely to prove acceptable to the equine industry, where specific genotypes are desired. Instead, to increase the number of stallions that can be effectively utilized for cryopreserved semen production, it is likely that more than one method for cryopreserving sperm will be necessary. This manuscript reviews some of the processes involved in cryopreservation, how individual sperm physiology affects the ability to survive freezing and thawing, and how cryopreservation protocols can be customized to maximize sperm cryosurvival on an individual stallion basis.
Publication Date: 2007-11-26 PubMed ID: 18178040DOI: 10.1016/j.anireprosci.2007.11.010Google Scholar: Lookup
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Summary

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This research examines the differences in survival rates of frozen stallion sperm, highlighting that different cryopreservation methods might be necessary to optimize results for individual stallions.

Cryopreservation of Stallion Semen

  • The process of freezing and storing sperm, known as cryopreservation, presents challenges specific to stallions.
  • Unlike the dairy industry, where bulls whose sperm does not cryopreserve well are replaced, such replacement is less acceptable in the equine industry where specific genetic types are preferred.

Stallion-to-Stallion Variability in Cryosurvival

  • The study recognizes significant variation in semen cryosurvival rates among different stallions, suggesting that a single cryopreservation method might not be effective universally.
  • Therefore, the study argues for customizing cryopreservation protocols depending on the specific physiological characteristics of individual stallions to maximize sperm survival rates.

Processes Involved in Cryopreservation

  • The paper also provides a review of some of the critical processes involved in cryopreservation, considering how specific physiological factors of individual sperm cells affect their survival rates when frozen and thawed.

Need for Multiple Cryopreservation Techniques in the Equine Industry

  • Due to the stallion-specific requirements for successful cryopreservation, the research concludes that different methods of cryopreservation may be necessary in the equine industry.
  • This approach aims at increasing the number of stallions utilized effectively in producing cryopreserved semen to maintain the desired genotypes.

Cite This Article

APA
Loomis PR, Graham JK. (2007). Commercial semen freezing: individual male variation in cryosurvival and the response of stallion sperm to customized freezing protocols. Anim Reprod Sci, 105(1-2), 119-128. https://doi.org/10.1016/j.anireprosci.2007.11.010

Publication

ISSN: 0378-4320
NlmUniqueID: 7807205
Country: Netherlands
Language: English
Volume: 105
Issue: 1-2
Pages: 119-128

Researcher Affiliations

Loomis, P R
  • Select Breeders Service, Inc., 1088 Nesbitt Road, Colora, MD 21917, USA. paulloomis@selectbreeders.com
Graham, J K

    MeSH Terms

    • Animals
    • Cryopreservation / methods
    • Cryopreservation / veterinary
    • Horses / physiology
    • Male
    • Semen Preservation / methods
    • Semen Preservation / veterinary
    • Spermatozoa / physiology

    Citations

    This article has been cited 35 times.
    1. 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
    2. Gutiérrez-Cepeda L, Crespo F, Blazquez JC, Serres C. Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation. Animals (Basel) 2023 Jan 22;13(3).
      doi: 10.3390/ani13030382pubmed: 36766271google scholar: lookup
    3. Akhtar MF, Ma Q, Li Y, Chai W, Zhang Z, Li L, Wang C. Effect of Sperm Cryopreservation in Farm Animals Using Nanotechnology. Animals (Basel) 2022 Sep 2;12(17).
      doi: 10.3390/ani12172277pubmed: 36077996google scholar: lookup
    4. 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
    5. Nikitkina EV, Dementieva NV, Shcherbakov YS, Atroshchenko MM, Kudinov AA, Samoylov OI, Pozovnikova MV, Dysin AP, Krutikova AA, Musidray AA, Mitrofanova OV, Plemyashov KV, Griffin DK, Romanov MN. Genome-wide association study for frozen-thawed sperm motility in stallions across various horse breeds. Anim Biosci 2022 Dec;35(12):1827-1838.
      doi: 10.5713/ab.21.0504pubmed: 35240017google scholar: lookup
    6. Catalán J, Yánez-Ortiz I, Tvarijonaviciute A, González-Arostegui LG, Rubio CP, Yeste M, Miró J, Barranco I. Impact of Seminal Plasma Antioxidants on Donkey Sperm Cryotolerance. Antioxidants (Basel) 2022 Feb 18;11(2).
      doi: 10.3390/antiox11020417pubmed: 35204299google scholar: lookup
    7. Orsolini MF, Meyers SA, Dini P. An Update on Semen Physiology, Technologies, and Selection Techniques for the Advancement of In Vitro Equine Embryo Production: Section II. Animals (Basel) 2021 Nov 20;11(11).
      doi: 10.3390/ani11113319pubmed: 34828049google scholar: lookup
    8. Madeja ZE, Podralska M, Nadel A, Pszczola M, Pawlak P, Rozwadowska N. Mitochondria Content and Activity Are Crucial Parameters for Bull Sperm Quality Evaluation. Antioxidants (Basel) 2021 Jul 27;10(8).
      doi: 10.3390/antiox10081204pubmed: 34439451google scholar: lookup
    9. Nikitkina E, Musidray A, Krutikova A, Anipchenko P, Plemyashov K, Shiryaev G. Efficiency of Tris-Based Extender Steridyl for Semen Cryopreservation in Stallions. Animals (Basel) 2020 Oct 4;10(10).
      doi: 10.3390/ani10101801pubmed: 33020383google scholar: lookup
    10. Indriastuti R, Ulum MF, Arifiantini RI, Purwantara B. Individual variation in fresh and frozen semen of Bali bulls (Bos sondaicus). Vet World 2020 May;13(5):840-846.
    11. Aurich J, Kuhl J, Tichy A, Aurich C. Efficiency of Semen Cryopreservation in Stallions. Animals (Basel) 2020 Jun 13;10(6).
      doi: 10.3390/ani10061033pubmed: 32545785google scholar: lookup
    12. Marzano G, Moscatelli N, Di Giacomo M, Martino NA, Lacalandra GM, Dell'Aquila ME, Maruccio G, Primiceri E, Chiriacò MS, Zara V, Ferramosca A. Centrifugation Force and Time Alter CASA Parameters and Oxidative Status of Cryopreserved Stallion Sperm. Biology (Basel) 2020 Jan 27;9(2).
      doi: 10.3390/biology9020022pubmed: 32012799google scholar: lookup
    13. Papas M, Catalán J, Fernandez-Fuertes B, Arroyo L, Bassols A, Miró J, Yeste M. Specific Activity of Superoxide Dismutase in Stallion Seminal Plasma Is Related to Sperm Cryotolerance. Antioxidants (Basel) 2019 Nov 9;8(11).
      doi: 10.3390/antiox8110539pubmed: 31717586google scholar: lookup
    14. Leemans B, Stout TAE, Soom AV, Gadella BM. pH-dependent effects of procaine on equine gamete activation†. Biol Reprod 2019 Nov 21;101(5):1056-1074.
      doi: 10.1093/biolre/ioz131pubmed: 31373616google scholar: lookup
    15. Alamaary MS, Haron AW, Ali M, Hiew MWH, Adamu L, Peter ID. Effects of four extenders on the quality of frozen semen in Arabian stallions. Vet World 2019 Jan;12(1):34-40.
      doi: 10.14202/vetworld.2019.34-40pubmed: 30936651google scholar: lookup
    16. Buss T, Aurich J, Aurich C. Evaluation of a portable device for assessment of motility in stallion semen. Reprod Domest Anim 2019 Mar;54(3):514-519.
      doi: 10.1111/rda.13390pubmed: 30592335google scholar: lookup
    17. Morrell JM, Johannisson A. Comparison of the Effect of Heterologous and Homologous Seminal Plasma on Motility and Chromatin Integrity of Stallion Spermatozoa Selected by Single Layer Centrifugation. J Vet Med 2014;2014:325451.
      doi: 10.1155/2014/325451pubmed: 26464926google scholar: lookup
    18. Blanch E, Tomás C, Hernández M, Roca J, Martínez EA, Vázquez JM, Mocé E. Egg yolk and glycerol requirements for freezing boar spermatozoa treated with methyl β-cyclodextrin or cholesterol-loaded cyclodextrin. J Reprod Dev 2014 Apr 24;60(2):143-9.
      doi: 10.1262/jrd.2013-073pubmed: 24492655google scholar: lookup
    19. Gil L, Galindo-Cardiel I, Malo C, González N, Alvarez C. Effect of Cholesterol and Equex-STM Addition to an Egg Yolk Extender on Pure Spanish Stallion Cryopreserved Sperm. ISRN Vet Sci 2013;2013:280143.
      doi: 10.1155/2013/280143pubmed: 24416597google scholar: lookup
    20. Gutiérrez-Cepeda L, Fernández A, Crespo F, Ramírez MÁ, Gosálvez J, Serres C. The effect of two pre-cryopreservation single layer colloidal centrifugation protocols in combination with different freezing extenders on the fragmentation dynamics of thawed equine sperm DNA. Acta Vet Scand 2012 Dec 5;54(1):72.
      doi: 10.1186/1751-0147-54-72pubmed: 23217215google scholar: lookup
    21. Bitton A, Frishling A, Kalo D, Roth Z, Arav A. The Impact of Precisely Controlled Pre-Freeze Cooling Rates on Post-Thaw Stallion Sperm. Animals (Basel) 2025 Dec 21;16(1).
      doi: 10.3390/ani16010021pubmed: 41514709google scholar: lookup
    22. Strassner FM, Demattio L, Siuda M, Malama E, Muffels G, Bollwein H. Relationships Between Metabolism of Cryopreserved Equine Sperm Determined by the Seahorse Analyzer and Sperm Characteristics Measured by Flow Cytometry and Computer-Assisted Analysis of Motility. Vet Sci 2025 Nov 21;12(12).
      doi: 10.3390/vetsci12121109pubmed: 41472089google scholar: lookup
    23. Ferrara MA, Preziosi G, Boni R, Ruggiero R, Gualandi SC. Quantitative holographic analysis in stallion spermatozoa following cryopreservation. Sci Rep 2025 Dec 4;15(1):43190.
      doi: 10.1038/s41598-025-24885-wpubmed: 41345140google scholar: lookup
    24. 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).
      doi: 10.3390/antiox14111363pubmed: 41300520google scholar: lookup
    25. 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
    26. Al-Kass Z, Morrell JM, Ntallaris T. Effect of Centrifugation of Stallion Semen Through a Low Density Colloid Prior to Freezing on Sperm Cryosurvival. Animals (Basel) 2025 Jun 25;15(13).
      doi: 10.3390/ani15131881pubmed: 40646780google scholar: lookup
    27. Zabala SM, Serres C, Montero N, Crespo F, Lorenzo PL, Pérez-Aguilera V, Oliet A, Hijón V, Moreno S, González-Zorn B, Gutiérrez-Cepeda L. Innovative Approaches to Avoid Antibiotic Use in Equine Semen Cryopreservation: Advancing Sustainable Reproductive Technologies. Animals (Basel) 2025 May 9;15(10).
      doi: 10.3390/ani15101368pubmed: 40427246google scholar: lookup
    28. Dayanıklı C, Bülbül B, Doğan Ş, Şengül E, Kırbaş M, Kal Y, Ataman MB. Improving Ram Semen Low Cryotolerance by Replacing the Seminal Plasma With That of High-Cryotolerant Rams or Extender. Vet Med Sci 2025 May;11(3):e70381.
      doi: 10.1002/vms3.70381pubmed: 40294117google scholar: lookup
    29. Baharun A, Pertiwi PPP, Rahmi A, Rachmadanti NC, Handarini R, Iskandar H, Samsudewa D, Maulana T, Said S, Darussalam I, Nurcholis N, Arifiantini RI. Frozen semen quality of Pasundan bulls with different individual variations. Open Vet J 2025 Jan;15(1):244-251.
      doi: 10.5455/OVJ.2025.v15.i1.22pubmed: 40092200google scholar: lookup
    30. Boni R, Ruggiero R, Di Palma T, Ferrara MA, Preziosi G, Cecchini Gualandi S. Stallion Sperm Freezing with Different Extenders: Role of Antioxidant Activity and Nitric Oxide Production. Animals (Basel) 2024 Aug 25;14(17).
      doi: 10.3390/ani14172465pubmed: 39272250google scholar: lookup
    31. Catalán J, Yánez-Ortiz I, Torres-Garrido M, Ribas-Maynou J, Llavanera M, Barranco I, Yeste M, Miró J. Impact of Seminal Plasma Antioxidants on DNA Fragmentation and Lipid Peroxidation of Frozen-Thawed Horse Sperm. Antioxidants (Basel) 2024 Mar 6;13(3).
      doi: 10.3390/antiox13030322pubmed: 38539855google scholar: lookup
    32. Dode MAN, Capobianco N, Vargas LN, Mion B, Kussano NR, Spricigo JF, Franco MM. Seminal cell-free DNA as a potential marker for in vitro fertility of Nellore bulls. J Assist Reprod Genet 2024 May;41(5):1357-1370.
      doi: 10.1007/s10815-024-03068-ypubmed: 38438770google scholar: lookup
    33. 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).
      doi: 10.3390/vetsci11020065pubmed: 38393083google scholar: lookup
    34. Seyedasgari F, Asadi B, Kim E. Seminal plasma modulates post-thaw longevity and motility of frozen sperm in dromedary camel. Anim Biosci 2023 Dec;36(12):1821-1830.
      doi: 10.5713/ab.23.0136pubmed: 37641835google scholar: lookup
    35. Ozimic S, Ban-Frangez H, Stimpfel M. Sperm Cryopreservation Today: Approaches, Efficiency, and Pitfalls. Curr Issues Mol Biol 2023 May 29;45(6):4716-4734.
      doi: 10.3390/cimb45060300pubmed: 37367049google scholar: lookup