RT info:eu-repo/semantics/article
T1 An optimized centrifugation protocol for ram sperm ensuring high sample yield, quality and fertility
A1 Neila Montero, Marta
A1 Fernández Riesco, Marta
A1 Montes Garrido, Rafael
A1 Palacín Martínez, Cristina
A1 Chamorro Álvarez, César Ángel
A1 Paz Cabello, Paulino de
A1 Álvarez García, Mercedes
A1 Anel López, Luis
A1 Anel Rodríguez, Luis
A2 Medicina y Cirugia Animal
K1 Veterinaria
K1 Artificial Insemination
K1 Cooled semen
K1 Ejaculate centrifugation
K1 Fertility
K1 Ovine
K1 Sperm handling
K1 3109 Ciencias Veterinarias
AB [EN] The optimization and implementation of artificial insemination (AI) in sheep is necessary to increase the livestock productivity through enhanced control of reproductive function. Sperm centrifugation is a common procedure in the ejaculate handling in AI and other assisted reproductive technologies (ART), as part of new methods of sperm analysis, selection or preservation. However, our research group previously established that this simple procedure might cause a large sperm loss and induce deleterious effects on the sperm function of the ovine species when high centrifugation forces are employed. To our knowledge, there are no studies on combined effect of extender and different centrifugal forces on ram sperm yield and quality. Furthermore, evidence of in vivo fertility rate using sperm obtained with various centrifugation forces is also lacking in this species. Thus, the objective of this work was to define the ideal conditions for ram semen centrifugation that will achieve the best quantity and quality sample to ensure unaffected fertilization ability of centrifuged ram sperm. The Experiment 1 evaluated the effect of the centrifugation procedure of two extenders (INRA 96 and Tyrode's) and two cooling protocols (Rapid and Slow Refrigeration 35 C to 15 Ce) on sperm recovery rate and quality (motility and kinetic parameters, viability, apoptosis and mitochondrial activity). INRA 96 combined with Slow Refrigeration and Tyrode's at room temperature registered the highest sperm recovery and quality values (P 0.05). In Experiment 2, the influence of three centrifugal forces (600, 1200 and 6000 g for 10 min) was assessed immediately after centrifugation on the technical performance and sperm functionality in diluted samples with INRA 96 and Tyrode's at the conditions set out in Experiment 1. The lowest pellet weight (P 0.05) without harmful effect on sperm physiological status (P > 0.05) was achieved at 1200 g, since 6000 g induced sperm motility damage (P 0.05) with both extenders. Finally, to ensure the total safety of the centrifugation protocol, Experiment 3 tested in a combined in vitro and in vivo test the effect of these three centrifugal forces on ram sperm quality after dilution (INRA 96) and liquid storage (6e8 h at 15 C). The damage produced by 6000 g on sperm motility (P 0.05) was maintained over time, coinciding with a lower fertility (P 0.05). In conclusion, ram sperm can be centrifuged in INRA 96 extender up to 1200 g for 10 min at 15 C as secure values with high recovery rates and without detrimental effects on sperm quality and fertility.
PB Elsevier Inc.
SN 0093-691X
LK https://hdl.handle.net/10612/18280
UL https://hdl.handle.net/10612/18280
NO Neila-Montero, M., Riesco, M. F., Montes-Garrido, R., Palacin-Martinez, C., Chamorro, C., de Paz, P., Alvarez, M., Anel, L., & Anel-Lopez, L. (2022). An optimized centrifugation protocol for ram sperm ensuring high sample yield, quality and fertility. Theriogenology, 191, 179-191. https://doi.org/10.1016/J.THERIOGENOLOGY.2022.08.006
DS BULERIA. Repositorio Institucional de la Universidad de León
RD Jul 11, 2024