Protocol optimization for sea bass sperm cryopreservation and assessment of post-thawing dilution to prolong sperm usefulness in aquaculture mediterranean species

• Main Authors: França, Thales S., González-López, Wendy A., Sánchez, Malbelys P., Pérez, Luz, Mañanós, Evaristo L., Felip, Alicia, Gómez Peris, A., Asturiano, Juan F.
• Other Authors: Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), European Commission, Generalitat Valenciana, Ministerio de Universidades (España), Conselho Nacional de Desenvolvimento Científico e Tecnológico (Brasil), Fundaçao Capes (Brasil)
• Format: Conference Object
• Language: unknown
• Published: 2022
• Subjects: Dicentrarchus labrax; Solea senegalensis; Anguilla anguilla; Aquaculture; European eel
• Online Access: http://hdl.handle.net/10261/310117

Resumen del trabajo presentado en el 8th International Workshop on the Biology of Fish Gametes, celebrado en Gdansk (Polonia) del 20 al 23 de septiembre de 2022. Our study aimed to optimize the sperm cryopreservation protocol of sea bass (Dicentrarchus labrax) and evaluate the post-thawing sperm dilution in species of interest for Mediterranean aquaculture. Sea bass sperm samples (n=6) were cryopreserved using a modified Non-Activating Media (NAM) extender with or without 5% Glucose (Glu), and 10% of Me2SO or MeOH as cryoprotectant. In a second experiment, sea bass sperm samples (n=7) were cryopreserved using a modified NAM extender and Me2SO. In both experiments, an aliquot of post-thawed sperm was diluted in NAM or NAM + Bovine Serum Albumin (BSA), or not diluted (control). Senegalese sole (Solea senegalensis) sperm samples (n=13) were cryopreserved. The post-thawed sperm was diluted in Mounib or NAM media, or not diluted (control). European eel (Anguilla anguilla) sperm samples (n=12) were cryopreserved. An aliquot of post-thaw sperm was diluted in P1 or P1 + BSA media, or not diluted (control). Kinetic parameters were checked after thawing: sea bass (0, 2.5 and 6 h), Senegalese sole (0 and 3 h), and European eel (0, 24, 48, 72 and 96 h). The total motility (MOT - %), progressive motility (MOTp - %), the velocities curvilinear (VCL - µm/s), straight line (VSL - µm/s) and average path (VAP - µm/s), were evaluated using a CASA-Mot software. The highest post-thawing kinetic parameters of sea bass sperm were observed when Me2SO or Me2SO+Glu were used and post-thawed sperm was diluted in NAM+BSA. The post-thaw sperm dilution increased kinetic parameters and prolonged sperm capacity compared to undiluted (control) samples. The post-thawing dilution did not increase the quality of the Senegalese sole sperm at 0 h. However, after 3 h, the spermatozoa diluted in NAM maintained the VCL. The European eel post-thaw sperm diluted in P1 maintained the MOT at 48 h and the velocities at 96 h after thawing. The combination ...