Cryopreservation of yellowtail kingfish (Seriola lalandi) sperm: Effects on metabolic enzymes and sperm quality

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Villalobos, Elias Figueroa ; Wilson, Rodolfo ; Pereira, Wellison Amorim ; Merino, Osvaldo ; Perez-Atehortua, Maritza ; Niedmann, Paola ; Avila, Sebastian ; Alvarado, Claudia ; Risopatron, Jennie ; Oliveira, Ricardo Pinheiro S. ; Farias, Jorge G. ; Isler, Ivan Valdebenito ; Villasante, Alejandro ; Romero, Jaime

• Tipo de documento: Artigo Científico
• Fonte: Aquaculture; v. 596, p. 9-pg., 2025-02-15.

Resumo

This study aimed to determine the effects of cryopreservation on sperm functionality and assess post-thaw sperm quality in yellowtail kingfish ( Seriola lalandi). Milt designated for cryopreservation was frozen in a solution containing Cortland (R) medium +1.3 M Me2SO + 0.2 M glucose +2 % BSA in a 1:3 ratio (milt:cryoprotectant). Fresh milt was used as the control. The samples were frozen in 0.5 mL straws using a programmable freezing system. After thawing, the samples were evaluated for motility using Computer-Assisted Sperm Analysis (CASA) with a phase-contrast optical microscope. Additionally, flow cytometry and a multimodal reader were employed to assess various sperm parameters, including plasma membrane integrity (SYBR-14/PI), mitochondrial membrane potential (Delta Psi MMit: JC-1), DNA fragmentation (TUNEL), ATP content (CellTiter-Glo (R)), intracellular calcium levels (Fluo-4 AM), lipid peroxidation (LPO), intracellular superoxide (DHE/SYTOX) and glutathione (GSH/ GSSG) production, activities of glutathione peroxidase (GPx), catalase (CAT) and 8-oxoguanine glycosylase (OGG1), and detection of 8-hydroxydeoxyguanosine (8-OHdG). In cryopreserved milt, significant differences in motility (64.14 +/- 2.2 %), plasma membrane integrity (61.5 +/- 2.7 %), mitochondrial membrane potential (55.4 +/- 2.4 %), DNA fragmentation (6.7 +/- 1.1 %), ATP content (7.5 +/- 1.5 nmol/109 spermatozoa), and intracellular calcium levels (59.8 +/- 2.5 nM/109 spermatozoa) were observed compared to fresh milt. Differences were also noted in LPO, O 2i- , GPx, GSH/GSSG, CAT, 8-OHdG and OGG1 between the cryopreserved and control groups. Additionally, correlations between sperm quality parameters, oxidative stress markers, and enzyme activity in cryopreserved spermatozoa were determined. These results suggest that cryopreservation induces significant alterations derived from oxidative damage in the functionality of yellowtail kingfish spermatozoa, and thus further research is necessary to explore the need for using antioxidants in the cryopreservation process of sperm from this species. (AU)

• Processo FAPESP: 18/25511-1 - Bioprospecção de bactérias probióticas bacteriocinogênicas: da otimização do cultivo à aplicação em sistemas de produção animal
• Beneficiário: Ricardo Pinheiro de Souza Oliveira
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 24/00151-3 - Aproveitamento de subproduto da extração de ágar de Gelidium spp. como ingrediente alternativo para alimentos aquáticos: Rumo à economia circular e à sustentabilidade
• Beneficiário: Wellison Amorim Pereira
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado