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  • Free amino acids are absorbed faster and assimilated more efficiently than protein in postlarval Senegal sole (Solea senegalensis)
    Publication . Rønnestad, I.; Conceição, L. E. C.; Aragão, C.; Dinis, Maria Teresa
    To improve the formulation of diets for the early stages of marine fish, assimilation rates of free amino acids (FAA) and protein in postlarval Senegal sole (Solea senegalensis) were determined. Fish (2.45 ± 0.87 mg dry weight) were tube fed 36 nL of a diet of FAA containing L-[35S] methionine (FAA diet) or bovine serum albumin, containing L-[methylated-14C]bovine serum albumin (Prot-diet), both at a concentration of 4.08 g/L. A time series was performed, and the amounts of label in incubation water, liver, gut and body carcass were quantified. The FAA diet was absorbed with a 3.5-times-higher transfer rate (P < 0.001) from the gut into the larval body tissues compared with the Prot-diet. The FAA diet also was assimilated with greater efficiency than the Prot-diet (80% versus 58%, P = 0.001). If we assume that the label present in the gut represents amino acids incorporated into the intestinal tissue, the assimilation efficiencies for the two diets were 89 and 64%. Therefore, FAA seems to be superior to protein as a dietary source of amino acids in Senegal sole postlarvae. However, because the absorption dynamics of protein and FAA differ, care should be taken when using the sources together to avoid amino acid imbalance.
  • Nutritional physiology during development of Senegalese sole (Solea senegalensis)
    Publication . Conceição, L. E. C.; Ribeiro, Laura; Engrola, S.; Aragão, C.; Morais, S.; Lacuisse, M.; Soares, Florbela; Dinis, Maria Teresa
    The Senegalese sole, a species with a complex metamorphosis, difficulties in weaning and with occasional problems of malpigmentation and skeletal deformities, is a good model species to study larval nutritional physiology. In addition, the early metamorphosis and acquisition of a peculiar non-proactive bottom-feeding behaviour make early weaning an important issue in sole hatcheries. The present work reviews recent findings in different aspects of nutritional physiology during the development of Senegalese sole, in an attempt to optimize the composition of sole diets and to understand what are the limiting factors for weaning sole. Both digestive enzymes activity and tracer studies using 14C-Artemia show that sole larvae, even at young stages, have a high capacity for digesting live preys. This is reflected in a high growth potential and low mortality rates for this species during the larval stage compared to other marine fish species. Based on the observation of the digestive enzymes profile, early introduction of inert microdiets in co-feeding with Artemia does not seem to affect intestinal function. However, when co-feeding is not provided, intestinal activity may be depressed. Furthermore, early introduction of microdiets in co-feeding with Artemia may have a positive effect on survival rates, but at the expense of lower growth rates and higher size dispersal. This may reflect variation in the adaptation capacity of individual larvae to inert diets. High dietary neutral lipid (soybean oil) content results in reduced growth and accumulation of lipid droplets in the enterocytes and affects the capacity of Senegalese sole larvae to absorb and metabolise dietary fatty acids (FA) and amino acids (AA). Through tube feeding of different 14C-lipids and free FA it has been shown that FA absorption efficiency increases with unsaturation and that sole larvae spare DHA from catabolism. In addition, it was demonstrated that absorption efficiency varies according to molecular form, being highest for free FA, lowest for triacylglycerols and intermediate for phospholipids. Live preys commonly used in larviculture do not seem to have a balanced AA profile for sole larvae. Furthermore, the ideal dietary AA composition probably changes during development. Rotifers and Artemia metanauplii are apparently deficient in one or more of the following AA depending on the larval development stage: histidine, sulphur AA, lysine, aromatic AA, threonine and arginine. It has also been demonstrated that balancing the dietary AA profile with dipeptides inArtemia-fed larvae increases AA retention and reduces AA catabolism. When supplementing larval diets with limiting AA it should also be considered that sole larvae have different absorption, and retention efficiencies for individual AA, and that they have the capacity to spare indispensable AA. In addition, the absorption of free AA is faster and more efficient than that of complex proteins. Improvements in biochemical composition of inert microdiets for sole are likely to contribute to the reproducible weaning success of Senegalese sole.
  • Dietary taurine supplementation enhances metamorphosis and growth potential of Solea senegalensis larvae
    Publication . Pinto, W.; Figueira, L.; Ribeiro, L.; Yúfera, M.; Dinis, Maria Teresa; Aragão, C.
    The effect of dietary taurine supplementation on growth performance, metamorphosis success and amino acid metabolism of Senegalese sole (Solea senegalensis) larvae was investigated. These parameters were assessed in larvae fed control and taurine supplemented microcapsules during the pelagic phase. Subsequently, a similar evaluation was carried out in newly settled larvae fed upon Artemia, in order to verify the effect of earlier dietary taurine supplementation in larvae reared under improved feeding conditions. Results showed that dietary taurine supplementation did not affect larval growth performance and metamorphosis during the pelagic phase. However, by the end of the trial, Senegalese sole previously fed taurine supplemented microcapsules had a significantly higher growth performance and metamorphosis completion success than larvae fed control microcapsules. These differences were likely related to the improvement of feeding conditions upon settlement, which probably helped revealing the positive effects of earlier dietary taurine supplementation on Senegalese sole performance. Additionally, Senegalese sole may have benefited from taurine antioxidant properties during metamorphosis, since larval antioxidant defences may saturate at this stage. Furthermore, results from metabolic trials have shown that dietary taurine supplementation significantly increased amino acid retention in Senegalese sole larvae when a concomitant increase of taurine body levels was found. Therefore, an increase in larval growth potential and metamorphosis success was observed under dietary taurine supplementation and these results may help understanding why dietary taurine supplementation has been reported to simultaneously increase taurine body levels and growth performance in other fish species, leading to a better comprehension on the role of taurine during fish development.
  • Is dietary taurine supplementation beneficial for gilthead seabream (Sparus aurata) larvae?
    Publication . Pinto, W.; Figueira, L.; Santos, A.; Barr, Y.; Helland, S.; Dinis, Maria Teresa; Aragão, C.
    This study evaluated the effect of dietary taurine supplementation on the growth performance and methionine metabolism of gilthead seabream larvae. For this purpose, a growth experiment was carried out in which seabream larvae were fed upon rotifers supplemented with “blank” or taurine-enriched liposomes. A complementary trial was also done in which seabream larvae fed with live prey were subsequently tube-fed a solution containing a L-[U-14C] methionine with or without a taurine supplement. Results from the growth experiment showed that rotifers were successfully enriched with taurine, but no effects were observed on larval growth performance, survival or amino acid composition. Furthermore, dietary taurine supplementation did not result in an increase of larval taurine levels, a factor that may have been determinant for the absence of effects observed on growth performance. In the tube-feeding trial, results showed that dietary taurine supplementation led to an increase of methionine retention in larvae. These findings suggest the existence of an active taurine biosynthesis pathway for gilthead seabream during the larval stage. Hence, gilthead seabream may not be dependent on dietary taurine to maintain the taurine body pool, since it may convert taurine from methionine if required. Taken together, the results from this study indicate that dietary taurine supplementation does not seem to enhance the larval growth performance in fish species able to biosynthesise taurine during this stage, which seems to be the case of gilthead seabream. However, this study also showed that in these species, dietary taurine supplementation may ultimately affect larval metabolism by increasing methionine availability for several important physiological purposes, contributing to a better understanding on the role of taurine during the early life stages of fish development.
  • Assimilation and catabolism of dispensable and indispensable free amino acids in post-larval Senegal sole (Solea senegalensis)
    Publication . Rønnestad, I.; Conceição, L. E. C.; Aragão, C.; Dinis, Maria Teresa
    The postprandial metabolism of dietary free amino acids (AA) was studied in post-larval Senegal sole, Solea senegalensis, aged 32 days after hatching (DAH). The diet was administered as a single pulse (36 nl, 43.1 mmol/l) using a micro tube-feeding technique and a dissolved mixture of crystalline AA. In four separate treatments the diet contained L [U-14C] tracer for two indispensable AA (IAA), lysine and arginine or two dispensable amino acids (DAA), glutamate and alanine. The post-larva absorbed all tested AA from the diet with similar efficiency (97.5%). A small fraction of the IAA was catabolised (11.5±1.1 and 15.1±3.3%, for lysine and arginine, respectively) and a high proportion was retained in the body (86.7±1.3 and 81.6±4.1%). For the DAA more were catabolised (64.9±5.3% and 41.4±7.2% for glutamate and alanine, respectively) and less were retained (32.9±5.1% and 56.3±7.2%). On this basis, it appears that post-larval Senegal sole use DAA in preference to IAA as energy substrates while the retention (assimilation efficiency) is better for the IAA. These results support other recent studies that early stages of fish have a better capacity to regulate AA catabolism than previously believed and that indispensable AA are saved for body growth.