Ultra-rapid freezing in spheres yields a higher cryoresistance than in straws but remains inferior to conventional slow freezing of stallion sperm

Edwin Francisco Larriva González; Jefferson Emmanuel Erráez Guaicha; Karen Estefanía Torres Ordóñez; José Mauricio Duma Pauta; Diego Sebastián Larriva Salazar; Diego Andrés Galarza Lucero

doi:10.1016/j.cryobiol.2025.105303

Ultra-rapid freezing in spheres yields a higher cryoresistance than in straws but remains inferior to conventional slow freezing of stallion sperm

Edwin Francisco Larriva González (First Author)
, Jefferson Emmanuel Erráez Guaicha
, Karen Estefanía Torres Ordóñez
, José Mauricio Duma Pauta
, Diego Sebastián Larriva Salazar
, Diego Andrés Galarza Lucero (Last Author)

Facultad de Ciencias Agronomicas Universidad de Cuenca
Washington State University Department of Veterinary Clinical Sciences

Research output: Contribution to journal › Article › peer-review

Abstract

This study evaluated the cryoresistance of stallion sperm frozen by ultra-rapid (UR) methods using microspheres and straws or by the conventionally-slow (CS) method. Sixteen ejaculates from four stallions were each divided into three aliquots according to the freezing method: UR freezing in 30-μL spheres (UR-Spheres) by direct immersion in liquid nitrogen (LN₂); UR freezing in 0.25-mL straws (UR-Straws) by direct horizontal submersion in LN₂; and CS freezing in LN₂ vapors. Ultra-rapid freezing medium included 100 mM trehalose +1 % BSA, and the CS freezing medium contained 5 % dimethylformamide. Conventional-slow freezing yielded higher sperm cryoresistance than both UR freezing methods (P < 0.05). UR-Spheres produced higher values of sperm kinematic, viability, and acrosomal integrity than UR-Straws (P < 0.05). Sperm head area and perimeter in both UR freezing groups increased compared to fresh and CS frozen samples (P < 0.05). In conclusion, the UR-Spheres method produced greater cryoresistance of stallion semen than UR-Straw method.

Original language	English
Article number	105303
Pages (from-to)	1-3
Number of pages	3
Journal	Cryobiology
Volume	121
DOIs	https://doi.org/10.1016/j.cryobiol.2025.105303
State	E-pub ahead of print - 8 Sep 2025

Keywords

Spheres
Stallion sperm
Straws
Ultra-rapid freezing

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10.1016/j.cryobiol.2025.105303

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Larriva González, E. F., Erráez Guaicha, J. E., Torres Ordóñez, K. E., Duma Pauta, J. M., Larriva Salazar, D. S., & Galarza Lucero, D. A. (2025). Ultra-rapid freezing in spheres yields a higher cryoresistance than in straws but remains inferior to conventional slow freezing of stallion sperm. Cryobiology, 121, 1-3. Article 105303. Advance online publication. https://doi.org/10.1016/j.cryobiol.2025.105303

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title = "Ultra-rapid freezing in spheres yields a higher cryoresistance than in straws but remains inferior to conventional slow freezing of stallion sperm",

abstract = "This study evaluated the cryoresistance of stallion sperm frozen by ultra-rapid (UR) methods using microspheres and straws or by the conventionally-slow (CS) method. Sixteen ejaculates from four stallions were each divided into three aliquots according to the freezing method: UR freezing in 30-μL spheres (UR-Spheres) by direct immersion in liquid nitrogen (LN2); UR freezing in 0.25-mL straws (UR-Straws) by direct horizontal submersion in LN2; and CS freezing in LN2 vapors. Ultra-rapid freezing medium included 100 mM trehalose +1 \% BSA, and the CS freezing medium contained 5 \% dimethylformamide. Conventional-slow freezing yielded higher sperm cryoresistance than both UR freezing methods (P < 0.05). UR-Spheres produced higher values of sperm kinematic, viability, and acrosomal integrity than UR-Straws (P < 0.05). Sperm head area and perimeter in both UR freezing groups increased compared to fresh and CS frozen samples (P < 0.05). In conclusion, the UR-Spheres method produced greater cryoresistance of stallion semen than UR-Straw method.",

keywords = "Spheres, Stallion sperm, Straws, Ultra-rapid freezing, Stallion sperm, Ultra-rapid freezing, Spheres, Straws, Esperma de semental, Congelaci{\'o}n ultrarr{\'a}pida",

author = "\{Larriva Gonz{\'a}lez\}, \{Edwin Francisco\} and \{Err{\'a}ez Guaicha\}, \{Jefferson Emmanuel\} and \{Torres Ord{\'o}{\~n}ez\}, \{Karen Estefan{\'i}a\} and \{Duma Pauta\}, \{Jos{\'e} Mauricio\} and \{Larriva Salazar\}, \{Diego Sebasti{\'a}n\} and \{Galarza Lucero\}, \{Diego Andr{\'e}s\}",

note = "Publisher Copyright: {\textcopyright} 2025 Society for Cryobiology",

year = "2025",

month = sep,

day = "8",

doi = "10.1016/j.cryobiol.2025.105303",

language = "Ingl{\'e}s",

volume = "121",

pages = "1--3",

journal = "Cryobiology",

issn = "0011-2240",

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Ultra-rapid freezing in spheres yields a higher cryoresistance than in straws but remains inferior to conventional slow freezing of stallion sperm. / Larriva González, Edwin Francisco (First Author); Erráez Guaicha, Jefferson Emmanuel; Torres Ordóñez, Karen Estefanía et al.
In: Cryobiology, Vol. 121, 105303, 08.09.2025, p. 1-3.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Ultra-rapid freezing in spheres yields a higher cryoresistance than in straws but remains inferior to conventional slow freezing of stallion sperm

AU - Larriva González, Edwin Francisco

AU - Erráez Guaicha, Jefferson Emmanuel

AU - Torres Ordóñez, Karen Estefanía

AU - Duma Pauta, José Mauricio

AU - Larriva Salazar, Diego Sebastián

AU - Galarza Lucero, Diego Andrés

PY - 2025/9/8

Y1 - 2025/9/8

N2 - This study evaluated the cryoresistance of stallion sperm frozen by ultra-rapid (UR) methods using microspheres and straws or by the conventionally-slow (CS) method. Sixteen ejaculates from four stallions were each divided into three aliquots according to the freezing method: UR freezing in 30-μL spheres (UR-Spheres) by direct immersion in liquid nitrogen (LN2); UR freezing in 0.25-mL straws (UR-Straws) by direct horizontal submersion in LN2; and CS freezing in LN2 vapors. Ultra-rapid freezing medium included 100 mM trehalose +1 % BSA, and the CS freezing medium contained 5 % dimethylformamide. Conventional-slow freezing yielded higher sperm cryoresistance than both UR freezing methods (P < 0.05). UR-Spheres produced higher values of sperm kinematic, viability, and acrosomal integrity than UR-Straws (P < 0.05). Sperm head area and perimeter in both UR freezing groups increased compared to fresh and CS frozen samples (P < 0.05). In conclusion, the UR-Spheres method produced greater cryoresistance of stallion semen than UR-Straw method.

AB - This study evaluated the cryoresistance of stallion sperm frozen by ultra-rapid (UR) methods using microspheres and straws or by the conventionally-slow (CS) method. Sixteen ejaculates from four stallions were each divided into three aliquots according to the freezing method: UR freezing in 30-μL spheres (UR-Spheres) by direct immersion in liquid nitrogen (LN2); UR freezing in 0.25-mL straws (UR-Straws) by direct horizontal submersion in LN2; and CS freezing in LN2 vapors. Ultra-rapid freezing medium included 100 mM trehalose +1 % BSA, and the CS freezing medium contained 5 % dimethylformamide. Conventional-slow freezing yielded higher sperm cryoresistance than both UR freezing methods (P < 0.05). UR-Spheres produced higher values of sperm kinematic, viability, and acrosomal integrity than UR-Straws (P < 0.05). Sperm head area and perimeter in both UR freezing groups increased compared to fresh and CS frozen samples (P < 0.05). In conclusion, the UR-Spheres method produced greater cryoresistance of stallion semen than UR-Straw method.

KW - Spheres

KW - Stallion sperm

KW - Straws

KW - Ultra-rapid freezing

KW - Stallion sperm

KW - Ultra-rapid freezing

KW - Spheres

KW - Straws

KW - Esperma de semental

KW - Congelación ultrarrápida

UR - https://www.scopus.com/pages/publications/105015092285

UR - https://www.sciencedirect.com/science/article/pii/S0011224025001099

UR - https://pubmed.ncbi.nlm.nih.gov/40925083/

U2 - 10.1016/j.cryobiol.2025.105303

DO - 10.1016/j.cryobiol.2025.105303

M3 - Artículo

AN - SCOPUS:105015092285

SN - 0011-2240

VL - 121

SP - 1

EP - 3

JO - Cryobiology

JF - Cryobiology

M1 - 105303

ER -

Ultra-rapid freezing in spheres yields a higher cryoresistance than in straws but remains inferior to conventional slow freezing of stallion sperm

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Keywords

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