Body growth, mitochondrial enzymatic capacities and aspects of the antioxidant system and redox balance under calorie restriction in young turbot (Scophthalmus maximus, L.)

Calorie restriction (CR) without undernutrition has been found to enhance stress resistance and life span in endotherms and ectotherms. We investigated the effect of 30% reduction in food offering on growth, aerobic capacities and oxidative stress parameters in young turbot (Scophthalmus maximus, L.). No differences in body weight, length and hepatosomatic index between the ad libitum (AL)-fed and the CR group occurred after 55 days of diet application. Of the mitochondrial marker enzymes, only citrate synthase (CS) activity in the liver was reduced under CR, whereas muscle CS activity and cytochrome oxidase (CCO) activity in both tissues remained the same in both feeding groups. The concentration of reduced glutathione increased significantly during feeding in muscle of CR fish, resulting in a more reduced glutathione redox ratio (GSH/GSSG) compared with AL fish muscle. Thiobarbituric acid-reactive lipid hydroperoxides (lipid peroxidation) but not protein carbonyl content (protein oxidation) was significantly reduced in CR fish muscle. Liver oxidative stress parameters did not vary significantly between experimental feeding groups. We conclude that 30% CR over 8 weeks has no adverse effect on young turbot. On the contrary, CR supports a reduced tissue oxidation state and reduces accumulation of lipid peroxidation products in muscle at sustained muscular aerobic capacity.     

Optimizing Electrical Activation of Porcine Oocytes by Adjusting Pre‐ and Post‐Activation Mannitol Exposure Times

Modifying electrical activation conditions have been used to improve in vitro embryo production and development in pigs. However, there is insufficient information about correlations of porcine embryo development with oocyte pre‐ and post‐activation conditions. The purpose of this study was to compare the developmental rates of porcine oocytes subjected to different mannitol exposure times, either pre‐ or post‐electrical activation, and to elucidate the reason for the optimal mannitol exposure time. Mannitol exposure times around activation were adjusted as 0, 1, 2 or 3 min. Blastocyst development were checked on day 7. Exposure of oocytes to mannitol for 1 or 2 min before electrical activation produced significantly higher blastocyst rates than exposure for 0 or 3 min. There was no significant difference in blastocyst rates when activated oocytes were exposed to mannitol for 0, 1, 2 or 3 min after electrical activation. While exposure of oocytes to mannitol for 1 min pre‐ and 3 min post‐activation showed significantly higher blastocyst development than 0 min pre‐ and 0 min post‐activation. It also showed higher maintenance of normal oocyte morphology than exposure for 0 min pre‐ and 0 min post‐activation. In conclusion, exposure of oocytes to mannitol for 1 min pre‐ and 3 min post‐activation seems to be optimal for producing higher in vitro blastocyst development of porcine parthenogenetic embryos. The higher blastocyst development is correlated with higher maintenance of normal morphology in oocytes exposed to mannitol for 1 min pre‐ and 3 min post‐activation.     

Comparison of Conventional Freezing and Vitrification with Dimethylformamide and Ethylene Glycol for Cryopreservation of Ovine Embryos

The aim of this work was to evaluate the efficiency of the cryoprotectants dimethylformamide and ethylene glycol for cryopreservation of ovine embryos using vitrification and conventional freezing. The recovered embryos were distributed randomly in three treatment groups: Gr. 1: conventional freezing (n = 44), Gr. 2: vitrification with ethylene glycol (n = 39) and Gr. 3: vitrification with dimethylformamide (n = 38). Quality of fresh embryos in control group as well as of frozen and vitrified embryos was examined by three methodologies: staining with propidium iodide and Hoechst 33258 and evaluation under fluorescent microscopy, evaluation of re‐expansion and hatching rates after culture, and determination of apoptotic index with TUNEL technique. It was established that re‐expansion rate in all treatment groups was similar. In the same time, hatching rates were higher in Gr. 1 (40.5%) and Gr. 2 (35.3%) in comparison with Gr. 3 (15.5%, p < 0.05). The number of dead cells in vitrified embryos of Gr. 2 and Gr. 3 was higher (42.6 ± 26.2 and 63.2 ± 34.65, respectively) in comparison with Gr. 1 (conventional freezing, 10.1 ± 8.5, p < 0.05). Embryos vitrified with dimethylformamide included the same quality of apoptotic cells that Gr. 1 (conventional freezing) and fresh embryos. In conclusion, the dimethylformamide and ethylene glycol used as cryoprotectant to vitrify ovine embryos, in the concentrations and exposition time tested in this work, were not as efficient as the conventional freezing for cryopreservation of ovine embryos Thus, the conventional freezing with ethylene glycol was the most efficient method to cryopreserve ovine embryos in comparison with vitrification.     

Response of offshore cultivated Laminaria saccharina to hydrodynamic forcing in the North Sea

The aim of the presented investigation was to test the sensibility of macroalgal aquaculture in offshore wind farms in the North Sea and to find arguments for the choice of appropriate sites among the planned wind farms. Based on experience with an offshore aquaculture farm of Laminaria saccharina conducted in 2002, we assessed the maximum hydrodynamic forces affecting farmed algae by applying the model software “WaveLoad”. Drag measured in a towing tank was considerably higher on algae with a more ruffled margin and wider blade collected from sheltered environments than on flat and narrow farmed Laminaria despite comparable blade areas. Drag varied according to frond size, current velocity and acceleration reaction. Dislodgement of laminarian holdfasts and the forces necessary to break the stipe depended on blade length and surface area. Neither did our measured nor our calculated values of drag exceed those forces, provided the algae had been grown in a current > 1 m s^− 1. Even in storm conditions with maximum current velocities of 1.52 m s^− 1 and wave heights of up to 6.4 m can cultivated L. saccharina withstand the high energy environment.     

Aquaporin 9 is expressed in the epididymis of immature and mature pigs

Aquaporins (AQPs) are channel proteins that facilitate the transepithelial and bidirectional movement of water. AQP9 is an aquaporin that is expressed in the mammalian epididymis. This water transport contributes to epididymal sperm concentration. This study aimed to examine the morphology of epididymal epithelium in piglets and boars, as well as the expression and immunolocalization of AQP9. The piglets presented an epididymal epithelium in differentiation with principal, basal and apical cells. The cellular population of the epididymal epithelium in boars consisted of principal, basal, apical, clear and narrow cells. The migratory cells known as halo cells were observed in the epididymis of both piglets and boars. AQP9 expression presented differences between piglets and boars. Moderate intensity of AQP9 immunoreaction was observed in the apical border of the epididymal epithelium of the caput and cauda regions in the piglet epididymis. A moderate‐to‐intense reaction for AQP9 was observed in the nuclei of epithelial cells of the three epididymal regions in the boar epididymis. The region of the cauda epididymis showed reactivity for AQP9 also in the apical border of the epithelium. It is believed that the AQP9 is already functional in piglets at only 1 week of age and is more active, playing a pivotal role in the caput and cauda regions of the epididymis. Moreover, the intense AQP9 expression in the apical border of epithelial cells in the cauda region of the boar epididymis suggests a higher performance of AQP9 in this region, where sperm complete their maturation process, stored and concentrated.