Effects of the inclusion of microalgae spirulina (Spirulina platensis) in the diet of Gilthead Sea bream (Sparus aurata) on growth and metabolism

In gilthead seabream aquaculture, the feed supplies in the market is very expensive due to its high content of animal protein. In this respect, spiruline appears to be a valuable substitute to animal and vegetable protein. In this study we performed two experiments. The scope of the first one was to determine the effect of the inclusion of Spirulina platensis hydrolyzed on the physiological state and growth in juveniles of Sparus aurata. A total of 180 individuals were fed for 128 days with three different feeds: control diet, diet with 2% of hydrolyzed microalgae (Sp2), and diet with 4% of hydrolyzed microalgae (Sp4).The experimental groups were tested in triplicate (except control group that was in duplicate). Biometric parameters were registered every two or three weeks. At the end of the experiment blood samples were collected to analyze plasma metabolites. After this we tried to evaluate the anti-oxidant response in animals remained from the first experiment using a toxicological assay with sodium nitrite lasting three days. Fish were divided into control, Spi 2% and Spi 4%, all them with and without NaNO2. Even then, the plasma metabolites data were collected after 24h and 72h. At the end of the first experiment the administration of S. platensis appeared to have a negative impact on growth of S. aurata respect the control feed. Furthermore, the lactate content registered showed a significant difference between the control and the spiruline administration. In the second experiment the spiruline feed showed a glucose and a lactate content with significant differences after 72h of exposition to nitrites respect the control group due to the interaction between nitrites and treatment. S. platensis hydrolyzed 2% and 4% do not seems a good substitution for S. aurata both as a growth enhancer and improver of health metabolic pathways. Its role as a good antioxidant has not been confirmed in these experiments.

In vivo and in vitro effects of genistein as obesogenic/anti-obesogenic compound in the lipid metabolism of rainbow trout (Oncorhynchus mykiss)

Nowadays, soy is one of the most used ingredients in the formulation of fish feed, due to the ample market supply, lower market price, high protein concentration and favorable amino acid composition. Nevertheless, soybean meal products are rich and primary diet source of phytoestrogens, as genistein, which may have a potential negative impact on growth, hormonal regulation and lipid metabolism in fish. The principal aim of this study was to better understand in vivo and in vitro genistein’s effects on lipid metabolism of rainbow trout. In adipose tissue it was showed an unclear role of genistein on lipid metabolism in rainbow trout, and in liver an anti-obesogenic effect, with an up-regulation of autophagy-related genes LC3b (in adipose tissue) and ATG4b (in liver and adipose tissue), a down-regulation of apoptosis-related genes CASP3 (in adipose tissue) and CASP8 (in liver). An increase of VTG mRNA levels in liver was also observed. Genistein partially exerted these effects via estrogen- receptor dependent mechanism. In white muscle, genistein seemed to promote lipid turnover, up-regulating lipogenic (FAS and LXR) and lipolytic (HSL, PPARα and PPARβ) genes. It seemed that genistein could exert its lipolytic role via autophagic way (up-regulation of ATG4b and ATG12l), not through an apoptotic pathway (down-regulation of CASP3). The effects of genistein on lipid-metabolism and apoptosis-related genes in trout muscle were not dose-dependent, only on autophagy-related genes ATG4B and ATG12l. Moreover, a partial estrogenic activity of this phytoestrogen was also seen. Through in vitro analysis (MTT and ORO assay), instead, it was observed an anti-obesogenic effect of genistein on rainbow trout adipocytes, and this effect was not mediated by ERs. Both in vivo and in vitro, genistein exerted its effects in a dose-dependent manner.

Effects of photoperiod and feeding regimes on Neuropeptide Y (NPY) and Agouti-related Protein (AgRP) expression in central areas of senegalese sole, Solea senegalensis

In the present study, we have tried to expand our knowledge about the endocrine mechanisms that regulate feeding and growth in cultured fish, which could be relevant for the improvement of fish farming conditions and feeding strategies. In order to reach this goal, we have investigated some orexigenic hormones, Neuropeptide Y (NPY) and the paralogues of Agouti-related protein, (AgRP1, AgRP2) in Solea senegalensis, an important species for Mediterranean aquaculture. We focused on hormones synchronization to different feeding regimes (diurnal vs nocturnal and random feeding) and photoperiod (light-dark cycle vs constant darkness). Therefore, the achieved results could also be relevant from a chronobiological perspective. Solea senegalensis specimen were reared in two different photoperiods, i.e.LD Light-Dark conditions as well as in DD conditions (constant darkness) along with different feeding regimes (fed at ML, Med and RND times), so to determine if mRNA expression of orexigenic hormones (NPY, AgRP1 and AgRP2) are entrained by feeding time and/or photoperiod. Our results show an independence of npy mRNA expression from the feeding time and suggest an endogenous control of npy expression in telencephalon of sole, while in optice tectum, npy expression could be entrained by the light-dark cycle. Our results on Senegalese sole AgRP1 and AgRP2 showed the same pattern of expression, indicating that expression of AgRPs is related to photoperiod in optic tectum, instead to feeding time. However the involvement of AgRP1 and AgRP2 in feeding behaviour should not be discarded in sole, as further research will be carried out with specimens maintained under different fasting conditions. our results reinforce the role of the telencephalon as the main neural area involved in the neuroendocrine control of food intake in fish, where endogenous NPY rhythms have been found, while diencephalon statistical variations weren’t observed suggesting that this brain area could be less involved in the neuroendocrine control of food intake in fish than previously thought.