RESUMO
In semen cryopreservation, egg yolk is still widely used as a non-penetrating cryoprotectant. Much has been developed in the search for alternatives for this biological product. This work aimed to evaluate the processed egg yolk through ultracentrifugation and/or sonication in the cryopreservation of swine semen. Twenty-seven semen doses were purchased from a commercial boar stud and processed for cryopreservation using egg yolk lactose 11% (control) extender, processed using two different methods: high-speed centrifugation and sonication. Then, they were submitted to freeze-thawing protocol and were assessed for kinematic and cell structural parameters. Samples in which extenders underwent centrifugation had better results in velocity parameters, meanwhile those that only sonication was performed had poorest results in this parameter. The preservation of the membrane and mitochondria structure had better results when the diluent was only centrifuged in comparison with the other treatments. Therefore, centrifugation of extender containing egg yolk is important for better cryopreservation of swine semen.
Assuntos
Criopreservação/veterinária , Preservação do Sêmen/veterinária , Suínos/fisiologia , Animais , Centrifugação/métodos , Centrifugação/veterinária , Criopreservação/métodos , Crioprotetores , Gema de Ovo/química , Congelamento , Masculino , Preservação do Sêmen/métodos , Sonicação/métodos , Sonicação/veterinária , Motilidade dos Espermatozoides , Espermatozoides/citologiaRESUMO
The current study assessed a semen cryopreservation protocol in the Amazonian catfish Leiarius marmoratus, a freshwater fish, of rheophilic behavior, and of great importance for Brazilian fish farming. Eight males (nâ¯=â¯8) were stripped and the semen was cryopreserved if total motility in fresh semen was higher than 80%. The external cryoprotectant Trehalose was then diluted in Beltsvile Thawing Solution (BTS) extender in the following concentrations: 50, 100, 150, and 200â¯mM. Semen samples were diluted in the media (1:9 v/v) being tested, then frozen in a container with nitrogen vapor (dryshipper), and stored in liquid nitrogen at -196⯰C. Motility parameters assessed post-thawing were performed by CASA-system and sperm cell integrity analyses (membrane integrity, DNA integrity, and mitochondrial function) were performed through fluorescence microscopy. As a result, no significant statistical difference was observed between treatments, independently of Trehalose concentrations tested in the following post-thawing analysis: membrane integrity, DNA integrity, mitochondrial functionality, and sperm motility duration. As of total and progressive motilities, the treatment containing 50â¯mM trehalose (15.6 and 9.5%, respectively), exhibited inferior results when compared to treatments with 150â¯mM (22.9 and 17.7%, respectively) and 200â¯mM (31.4 and 26.3%, respectively) trehalose concentrations (Pâ¯<â¯0.05); however, it did not differ from the treatment with 100â¯mM trehalose (18.6 and 15.3%, respectively). Therefore, treatments with trehalose at higher concentrations exhibited superior results when compared to other treatments in in vitro motility parameters for L. marmoratus.