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- Characterization of spawning rhythms of a sparidae aquaculture species, the white seabream (Diplodus sargus), in the south of PortugalPublication . Santos, Cátia; Soares, Florbela; Candeias-Mendes, Ana; Pousão‐Ferreira, Pedro; Dinis, Maria Teresa; Oliveira, CatarinaOne of the most visible features of fish reproduction is seasonality; however, daily rhythmicity is present in most species and should not be disregarded. This study investigated the seasonal and daily spawning rhythms of the white seabream (Diplodus sargus) in captivity, and a possible endogenous control of the daily rhythm. Spawns were monitored daily, during an entire reproductive season by using an automatic and programmable egg collector. The spawning season lasted 5 months, from January until June, when temperatures oscillated between 11.5 and 24.4 degrees C. Female fecundity was higher between 17.5 and 20.4 degrees C, suggesting a temperature influence in spawning. This species showed a very clear daily spawning rhythm (Cosinor analysis, p < 0.001), strongly related with photoperiod. Eggs started to be released at sunset and peaked around 21:00 h. Egg quality parameters also changed along the spawning period, with viability and hatching rates, increasing towards the end of the night period. The spawning rhythm persisted (Cosinor analysis, p < 0.001) for 5 days under constant light conditions, suggesting the presence of an endogenous clock mechanism which controlled this circadian pattern. The present results represent an advantage for the optimization of egg production protocols in this species, and an added value to the current knowledge of marine fish physiology.
- Solea senegalensis sperm cryopreservation: new insights on sperm qualityPublication . F RIESCO, MARTA; Oliveira, Catarina; Soares, Florbela; Gavaia, Paulo J.; Dinis, Maria Teresa; Cabrita, ElsaCryopreservation of Senegalese sole sperm can represent an alternative to overcome some reproductive problems of this species. However, it is important to guarantee the safe use of cryopreserved sperm by selecting an appropriate protocol according to a high demand quality need to be ensured. It has been demonstrated that traditional assays such as motility and viability do not provide enough information to identify specific damage caused by cryopreservation process (freezing and thawing). Specific tests, including lipid peroxidation and DNA damage, should be performed. In the present study, motility and lipid peroxidation were performed as specific tests allowing us to discard cryopreservation conditions such as methanol as internal cryoprotectant and bovine serum albumin as external cryoprotectant. In addition, a caspase 3/7 detection by flow cytometry was performed to analyze apoptosis activity in the best selected conditions. Moreover, new highly sensitive tests based on transcript number detection have recently been described in fish sperm cryopreservation. For this reason, a transcript level detection assay was performed on certain oxidative and chaperone genes related to fertilization ability and embryo development (hsp70, hsp90BB, hsp90AA, gpx) to select the best cryopreservation conditions. DMSO+ egg yolk proved to be the best cryoprotectant combination in terms of transcript level. This study describes an optimized cryopreservation protocol for Solea senegalensis sperm demonstrating for the first time that transcript degradation is the most sensitive predictor of cell status in this species after cryopreservation.
- Optimizing rearing and welfare in Senegalese sole (Solea senegalesensis) broodstock: effect of ambient light intensity and handling time on stress responsePublication . Figueiredo, Filipe; Aragão, Cláudia; Pinto, Wilson; Dinis, Maria Teresa; Oliveira, CatarinaBroodstock rearing conditions and handling procedures should be optimized in aquaculture species in order to benefit fish welfare and guarantee optimal conditions for spawning. In teleosts, basal cortisol levels display daily rhythms, oscillating along the 24 h of the day. In this sense, handling fish at different moments of the day may lead to different stress responses. The present study aimed at investigating the optimal rearing conditions for Senegalese sole broodstock, considering ambient light intensity and handling time. The optimal light intensity (50, 100 or 200 lx) was investigated by measuring fish cortisol levels and monitoring locomotor activity rhythms under each intensity tested. Results showed a significant increase in cortisol levels of fish exposed to 200 lx, when compared to values obtained under 100 lx, accompanied by changes in locomotor activity rhythms in both tanks under study. These results suggested that 200 lx may be too high as light intensity for this species, whereas 100 lx seems to be more adequate. Also, daily rhythms of stress response were investigated in breeders from different origins (Wild and first generation, G1). Basal cortisol levels and cortisol stress response after an acute stressor (air exposure) were monitored at two distinct moments of the day (Mid-Light and Mid-Dark). Basal levels were higher during the day in the wild group, while G1 fish seemed to have lost the daily fluctuations in basal cortisol plasma levels, as well as their daily rhythms of locomotor activity. Both groups showed lower stress responses during night-time, an indication that this is an adequate period of the day to handle this species. Senegalese sole breeders born in captivity presented more pronounced stress responses when compared to wild fish, reflecting their different life history in terms of stress challenges.