Influence of extender type, freezing temperature, and sperm concentration on the cryosurvival of Santa Inês ram spermatozoa
6 affiliations
- 1Department of Animal Science, Center for Agricultural and Biological Sciences, State University Vale do Acaraú (UVA), Sobral, Ceará Brazil
- 2Department of Animal Science, “Luiz de Queiroz” College of Agriculture (ESALQ), University of São Paulo (USP), Piracicaba, São Paulo, Brazil
- 3Center for Education, Sciences and Technology of the Inhamuns Region, State University of Ceará (UECE), Tauá, Ceará Brazil
- 4Department of Animal Science, Center for Agricultural Sciences, Federal University of Ceará (UFC), Fortaleza, Ceará Brazil
- 5Federal Institute of Education, Science and Technology of Ceará (IFCE), Umirim Campus, Umirim, Ceará Brazil
- 6School of Veterinary Medicine, State University of Ceará (UECE), Fortaleza, Ceará Brazil
Abstract
This study evaluated the effects of three extenders (Tris, HEPES, and skim milk), three freezing temperatures (-79, -90, and − 120 °C), and three final sperm concentrations (600, 800, and 1000 × 10⁶ spermatozoa/mL) on the post-thaw quality of Santa Inês ram semen. Semen was collected weekly from five rams during six consecutive weeks, pooled, and subjected to a 3 × 3 × 3 factorial arrangement. Samples were cryopreserved using a programmable freezing system (TK3000®) and stored in liquid nitrogen. After thawing, sperm quality was assessed by computer-assisted sperm analysis (CASA) and sperm morphology evaluation. Data were analyzed using mixed models, considering extender, freezing temperature, sperm concentration, and their interactions as fixed effects and collection week as a random effect. Tris consistently resulted in greater (P < 0.05) total motility, progressive motility, and percentage of rapid spermatozoa compared with HEPES and skim milk extenders. Semen frozen at 600 × 10⁶ spermatozoa/mL showed superior post-thaw motility characteristics relative to higher sperm concentrations. Freezing temperature affected sperm kinetics, particularly in skim milk and HEPES extenders, whereas Tris maintained stable performance across the evaluated temperatures. Total morphological abnormalities were minimally influenced by the treatments. Overall, under the experimental conditions evaluated, the combination of Tris extender, a freezing temperature of -79 °C, and a sperm concentration of 600 × 10⁶ spermatozoa/mL provided the most favorable post-thaw sperm quality among the treatments tested. However, progressive motility remained relatively low, indicating that further optimization and fertility validation are required before this combination can be recommended for practical application.
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BibTeX
@article{lima2026influence,
title = {{Influence of extender type, freezing temperature, and sperm concentration on the cryosurvival of Santa Inês ram spermatozoa}},
author = {Fátima Révia Granja Lima and Paula Renata Cortat and Robson Mateus Freitas Silveira and Mirela Balistrieri and Renato Mesquita Peixoto and Ana Glaudia Catunda and Ana Claudia Nascimento Campos and Airtor Alencar Araújo},
journal = {Tropical Animal Health and Production},
year = {2026},
month = nov,
doi = {10.1007/s11250-026-05435-6},
url = {https://doi.org/10.1007/s11250-026-05435-6},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Influence of extender type, freezing temperature, and sperm concentration on the cryosurvival of Santa Inês ram spermatozoa
AU - Fátima Révia Granja Lima
AU - Paula Renata Cortat
AU - Robson Mateus Freitas Silveira
AU - Mirela Balistrieri
AU - Renato Mesquita Peixoto
AU - Ana Glaudia Catunda
AU - Ana Claudia Nascimento Campos
AU - Airtor Alencar Araújo
PY - 2026
DA - 2026/11/11
JO - Tropical Animal Health and Production
DO - 10.1007/s11250-026-05435-6
UR - https://doi.org/10.1007/s11250-026-05435-6
AB - This study evaluated the effects of three extenders (Tris, HEPES, and skim milk), three freezing temperatures (-79, -90, and − 120 °C), and three final sperm concentrations (600, 800, and 1000 × 10⁶ spermatozoa/mL) on the post-thaw quality of Santa Inês ram semen. Semen was collected weekly from five rams during six consecutive weeks, pooled, and subjected to a 3 × 3 × 3 factorial arrangement. Samples were cryopreserved using a programmable freezing system (TK3000®) and stored in liquid nitrogen. After thawing, sperm quality was assessed by computer-assisted sperm analysis (CASA) and sperm morphology evaluation. Data were analyzed using mixed models, considering extender, freezing temperature, sperm concentration, and their interactions as fixed effects and collection week as a random effect. Tris consistently resulted in greater (P < 0.05) total motility, progressive motility, and percentage of rapid spermatozoa compared with HEPES and skim milk extenders. Semen frozen at 600 × 10⁶ spermatozoa/mL showed superior post-thaw motility characteristics relative to higher sperm concentrations. Freezing temperature affected sperm kinetics, particularly in skim milk and HEPES extenders, whereas Tris maintained stable performance across the evaluated temperatures. Total morphological abnormalities were minimally influenced by the treatments. Overall, under the experimental conditions evaluated, the combination of Tris extender, a freezing temperature of -79 °C, and a sperm concentration of 600 × 10⁶ spermatozoa/mL provided the most favorable post-thaw sperm quality among the treatments tested. However, progressive motility remained relatively low, indicating that further optimization and fertility validation are required before this combination can be recommended for practical application.
SN - 0049-4747
SN - 1573-7438
ER -