RT info:eu-repo/semantics/article
T1 Effect of basic factors of extender composition on post-thawing quality of brown bear electroejaculated spermatozoa
A1 Anel Rodríguez, Luis
A1 Gomes Alves, Susana Cláudia
A1 Álvarez García, Mercedes
A1 Barragán Santos, Santiago
A1 Anel Rodríguez, Enrique
A1 Nicolás, M.
A1 Martínez Pastor, Felipe
A1 Paz Cabello, Paulino de
A2 Biologia Celular
K1 Veterinaria
K1 Brown bear
K1 Cryopreservation
K1 Spermatozoa
K1 Extender
K1 Germplasm banks
AB The improvement of freezing extenders is critical when defining sperm cryopreservation protocols for wild species, in order to create germplasm banks. The aim of this study was to evaluate the effect of additives (Equex Paste and EDTA) supplementation, egg-yolk (10 and 20%) and glycerol (4 and 8%) concentrations and extender osmolality (300 and 320 mOsm/kg) on the post-thawing quality of brown bear semen. Semen was obtained from 20 adult males by electroejaculation, and centrifugated individually (600 × g for 6 min). The pellets were diluted 1:1 in the corresponding extender TTF (TES-Tris-Fructose with the aforementioned variants) and cooled to 5 °C. Then, it was diluted down to 100 × 106 spz/mL, loaded in 0.25 mL straws and frozen at −20°C/min. After thawing (in water at 65 °C for 6s), the semen samples were assessed for motility (CASA), viability (SYBR-14 with propidium iodide), acrosomal status (PNA-FITC with propidium iodide) and mitochondrial activity (JC-1). Extender supplementation with additives rendered significantly higher results for these sperm parameters. Comparing the two percentages of egg yolk, 20% egg yolk showed the highest motility results, percentages of viable spermatozoa and viable spermatozoa with intact acrosome. No differences were detected among samples frozen using 4 or 8% glycerol. For extender osmolality, 300 mOsm/kg showed higher values of VAP, VCL, VSL, and ALH than 320 mOsm/kg. Based on the best performance of sperm motility, viability and acrosome status, we conclude that the most suitable extender to cryopreserve brown bear spermatozoa was TTF adjusted to 300 mOsm/kg, supplemented with 20% egg yolk, 4–8% glycerol, and the additives 1% Equex paste and 2% EDTA.
PB Elsevier
YR 2019
FD 2019-05-13
LK http://hdl.handle.net/10612/10744
UL http://hdl.handle.net/10612/10744
NO Theriogenology, 2010, vol. 74, n. 4
NO P. 643-651
DS BULERIA. Repositorio Institucional de la Universidad de León
RD Jul 6, 2024