RESUMO
BACKGROUND: The assessment of oocyte quality is, nowadays, a major challenge in aquaculture, oocyte cryopreservation, and environmental science. Oocyte quality is a determining factor in fertilization and embryo development; however, there is still a lack of rapid and sensitive cellular markers for its assessment. Currently, its estimation is predominantly based on morphological analysis, which is subjective and does not consistently reflect the developmental competence of the oocytes. Despite several recent studies investigating molecular markers related to oocyte quality, methods currently available for their determination pose various technical challenges and limitations. In this study, we developed a novel approach based on fluorescence spectroscopy to assess different intrinsic physiological parameters that can be employed to evaluate egg quality in marine invertebrates that are widely used as animal models such as sea urchins and mussels. RESULTS: Different physiological parameters, such as viability, mitochondrial activity, intracellular ROS levels, plasma membrane lipid peroxidation, and intracellular pH, for egg quality evaluation have been successfully assessed in sea urchins and mussels by using specific fluorescent dyes and detecting the fluorescent signals in eggs through fluorescence spectroscopy. CONCLUSIONS: Based on our findings, we propose these physiological markers as useful predictors of egg quality in marine invertebrates; they can be estimated rapidly, selectively, and sensitively by employing this novel approach, which, due to the speed of analysis, the low cost, and easy use can be considered a powerful analytical tool for the egg quality assessment.
Assuntos
Desenvolvimento Embrionário , Oócitos , Animais , Espectrometria de Fluorescência , Oócitos/metabolismo , Ouriços-do-Mar , Criopreservação/métodosRESUMO
BACKGROUND: The assessment of oocyte quality is, nowadays, a major challenge in aquaculture, oocyte cryopreservation, and environmental science. Oocyte quality is a determining factor in fertilization and embryo development; however, there is still a lack of rapid and sensitive cellular markers for its assessment. Currently, its estimation is pre-dominantly based on morphological analysis, which is subjective and does not consistently reflect the developmental competence of the oocytes. Despite several recent studies investigating molecular markers related to oocyte quality, methods currently available for their determination pose various technical challenges and limitations. In this study, we developed a novel approach based on fluorescence spectroscopy to assess different intrinsic physiological parameters that can be employed to evaluate egg quality in marine invertebrates that are widely used as animal models such as sea urchins and mussels. RESULTS: Different physiological parameters, such as viability, mitochondrial activity, intracellular ROS levels, plasma membrane lipid peroxidation, and intracellular pH, for egg quality evaluation have been successfully assessed in sea urchins and mussels by using specific fluorescent dyes and detecting the fluorescent signals in eggs through fluorescence spectroscopy. CONCLUSIONS: Based on our findings, we propose these physiological markers as useful predictors of egg quality in marine invertebrates; they can be estimated rapidly, selectively, and sensitively by employing this novel approach, which, due to the speed of analysis, the low cost, and easy use can be considered a powerful analytical tool for the egg quality assessment.
Assuntos
Animais , Oócitos/metabolismo , Desenvolvimento Embrionário , Ouriços-do-Mar , Espectrometria de Fluorescência , Criopreservação/métodosRESUMO
In the development of functional probiotic food, the carrier matrices should be carefully selected and optimized to ensure the highest levels of probiotic survival in the symbiotic food along storage. Because milk and honey food matrices are rich in antioxidant substances, the aim of the research was to evaluate their effect in protecting lactobacilli from reactive oxygen species (ROS) generated by the addition of hydrogen peroxide. Viability assays were performed with and without the addition of H2O2, in three different matrices: 0.9% peptone saline, 5% honey, or 12% reconstituted skim milk. The milk matrix provided protection for the Lacticaseibacillus paracasei DTA83 and Lacticaseibacillus rhamnosus DTA76. However, this protective effect was not observed in the survival of Lactobacillus acidophilus La 5. Honey solution did not maintain the viability of probiotic microorganisms exposed to hydrogen peroxide and, on the contrary, caused a significant reduction in the population of L. rhamnosus DTA76 (p < 0.001). Lower membrane lipid peroxidation due to H2O2 exposure was observed in L. acidophilus La 5 and L. rhamnosus DTA76, but this marker showed no relation with viability. It was concluded: (i) lactobacilli from the Lacticaseibacillus genus were the ones that benefited most from the lactic environment; (ii) the absence of the protective effect of honey was possibly due to the presence of Fe2+ which reacts with H2O2 to produce hydroxyl radicals; and (iii) cell viability did not correlate with membrane lipid peroxidation, and it is not a good marker to evaluate this type of damage in cells of different microorganisms.