Influence of sperm density on cryopreservation outcomes in sturgeon

• Autor(a) principal: Nascimento, João Pedro Oliveira
• Data de Publicação: 2018
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10400.1/12579
• topic: Qualidade do esperma; Concentração; Ploidismo; Mobilidade; Criopreservação; Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias

The goals of this experiment were the establishment of procedure for variation of spermatozoid concentration without compromising quality, establishment of highest cell concentration without decreasing post-thaw motility by routine freezing protocol and influence of ploidy level. It was used sperm from sterlet sturgeon (Acipenser ruthenus, n=6), Siberian sturgeon (Acipenser baerii, n=6) and from Russian sturgeon (Acipenser gueldenstaedii, n=4). It was tested a method of increasing concentration of spermatozoa by centrifuging with the following conditions (300 x g, 10 min, 4ºC and 300 x g, 20 min, 4ºC). To find the ideal concentration, it was tested 4 dilutions (1:1, 1:4, 1:9 and 1:49) and 2 controls (one control with spermatozoa without being centrifuged and another control with concentrated spermatozoa without the addition of seminal fluid). To perform these 4 dilutions, the concentrated spermatozoa was diluted with seminal fluid (SF). Then, the sperm was diluted in a cryoprotective medium containing 10% methanol and frozen by a conventional freezing protocol. It was measured the concentration of spermatozoa and motility parameters. It was determined the critical concentration of sperm in the straw where it was known that relative motility was 50% by performing a simple regression line. Spermatozoa after centrifugation had no changes in its quality and it was possible to increase concentration up to 10 times the initial concentration. For post-thaw spermatozoa, the highest motilities were obtained in samples with concentrations lower than 1 x 109 spz mL-1 for all three species (57.5 % for sterlet sturgeon, 40% for Siberian sturgeon and 20% for Russian sturgeon). The critical concentrations in the straw for when we know the relative post-thaw motility is 50% was 1.05 x 109 spz mL-1 for sterlet, 0.92 x 109 spz mL-1 for Siberian sturgeon and for Russian sturgeon was 1.17 x 109 spz mL-1. Regarding motility parameters, the concentrations lower than the critic level had higher VCL values (164 μm s-1 for sterlet sturgeon, 179.26 μm s-1 for Siberian sturgeon and 145.48 μm s-1 for Russian sturgeon). There was significant difference (p < 0.05) when comparing the velocities between the spermatozoa with concentration lower than critic level and higher than critic level in sterlet sturgeon and Siberian sturgeon but not in Russian sturgeon. Our results show a dependency of the concentration and spermatozoa size in cryopreservation outcomes.