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- Nutrition, quality and performance in marine fish larvaePublication . Conceição, L. E. C.; Alves-Martins, D.; Engrola, S.; Gavaia, Paulo J.; Richard, Nadège; Aragão, C.O cultivo de larvas de peixes marinhos é caracterizado por altas mortalidades e por problemas de qualidade, nomeadamente malformações esqueléticas. A solução de parte destes problemas passa por uma melhor compreensão dos requisitos nutricionais. Este trabalho pretende fazer uma revisão de trabalhos recentes que demonstram como os aminoácidos, os ácidos gordos e a vitamina K, mas também o regime alimentar, podem influenciar a qualidade e a performance das larvas de peixes marinhos.
- Novel methodologies in marine fish larval nutritionPublication . Conceição, L. E. C.; Aragão, C.; Richard, Nadège; Engrola, S.; Gavaia, Paulo J.; Mira, Sara; Dias, J.Major gaps in knowledge on fish larval nutritional requirements still remain. Small larval size, and difficulties in acceptance of inert microdiets, makes progress slow and cumbersome. This lack of knowledge in fish larval nutritional requirements is one of the causes of high mortalities and quality problems commonly observed in marine larviculture. In recent years, several novel methodologies have contributed to significant progress in fish larval nutrition. Others are emerging and are likely to bring further insight into larval nutritional physiology and requirements. This paper reviews a range of new tools and some examples of their present use, as well as potential future applications in the study of fish larvae nutrition. Tube-feeding and incorporation into Artemia of 14C-amino acids and lipids allowed studying Artemia intake, digestion and absorption and utilisation of these nutrients. Diet selection by fish larvae has been studied with diets containing different natural stable isotope signatures or diets where different rare metal oxides were added. Mechanistic modelling has been used as a tool to integrate existing knowledge and reveal gaps, and also to better understand results obtained in tracer studies. Population genomics may assist in assessing genotype effects on nutritional requirements, by using progeny testing in fish reared in the same tanks, and also in identifying QTLs for larval stages. Functional genomics and proteomics enable the study of gene and protein expression under various dietary conditions, and thereby identify the metabolic pathways which are affected by a given nutrient. Promising results were obtained using the metabolic programming concept in early life to facilitate utilisation of certain nutrients at later stages. All together, these methodologies have made decisive contributions, and are expected to do even more in the near future, to build a knowledge basis for development of optimised diets and feeding regimes for different species of larval fish.
- Changes in plasma amino acid levels in a euryhaline fish exposed to different environmental salinitiesPublication . Aragão, C.; Costas, B.; Vargas-Chacoff, L.; Ruiz-Jarabo, I.; Dinis, Maria Teresa; Mancera, J. M.; Conceição, L. E. C.Previous studies have shown that Senegalese sole is partially euryhaline in the juvenile phase, being able to adapt to a wide range of salinities in a short-time period, due to changes at the osmoregulatory and metabolic level. This study aimed to assess the effects of acclimation of sole to a wide range of salinities, with a special emphasis on the role of plasma amino acids during this process. Sole juveniles were acclimated for 2 weeks to different salinities: 5, 15, 25, 38, and 55 g L−1. Plasma levels of cortisol, glucose, osmolality, and free amino acids were assessed at the end. Changes in plasma levels of cortisol, glucose, and amino acids indicate that fish reared at 5 and 55 g L−1 were facing extra energy costs. Amino acids seem to play an important role during salinity acclimation, either as energy sources or as important osmolytes for cell volume regulation.
- Can dietary aromatic amino acid supplementation be beneficial during fish metamorphosis?Publication . Pinto, W.; Rodrigues, V.; Dinis, Maria Teresa; Aragão, C.Aromatic amino acids (AA, phenylalanine and tyrosine) are the precursors of thyroid hormones, which regulate metamorphosis in all vertebrates. In fish, this is a critical developmental stage where physiological requirements for aromatic AA may increase. Therefore, it is possible that dietary aromatic AA supplementation may be beneficial to accomplish a successful transition from larvae to the juvenile stage. This work aimed to assess the effect of dietary aromatic AA supplementation on tyrosine metabolism in species with an accentuated (Senegalese sole, Solea senegalensis) and a less marked (gilthead seabream, Sparus aurata) metamorphosis. For this purpose, either species were fed Artemia and subsequently received or not an aromatic AA supplement through tube-feeding, together with a L-[U-14C] tyrosine marker, throughout metamorphosis. Tyrosine was elected as a tracer since the metabolic fate of this aromatic AA is directly linked to the production of thyroid hormones and metamorphosis. Results showed that dietary phenylalanine supplementation did not increase tyrosine retention in Senegalese sole larvae during metamorphosis, suggesting that this species may not be able to biosynthesise tyrosine from phenylalanine at a sufficient rate to supply its physiological requirements until after metamorphosis. Furthermore, dietary tyrosine supplementation effectively increased tyrosine availability in Senegalese sole body fluids at metamorphosis, most likely for coping with metamorphosis-related processes, such as production of thyroid hormones. Therefore, dietary tyrosine supplementation may be beneficial for Senegalese sole during metamorphosis. On the other hand, results indicated that gilthead seabream larvae do not seem to require an additional supply of aromatic AA during metamorphosis, probably because the physiological requirements for tyrosine do not increase during this developmental stage. The different results observed for Senegalese sole and gilthead seabream are probably related to the complexity of metamorphosis that each species undergoes and to the needs for production of thyroid hormones, which seems to affect aromatic AA requirements during this critical stage of development. These findings may be important for physiologists, fish nutritionists and for the flatfish aquaculture industry.
- Dietary nitrogen and fish welfarePublication . Conceição, L. E. C.; Aragão, C.; Dias, J.; Costas, B.; Terova, G.; Martins, C.; Tort, L.Little research has been done in optimizing the nitrogenous fraction of the fish diets in order to minimize welfare problems. The purpose of this review is to give an overview on how amino acid (AA) metabolism may be affected when fish are under stress and the possible effects on fish welfare when sub-optimal dietary nitrogen formulations are used to feed fish. In addition, it intends to evaluate the current possibilities, and future prospects, of using improved dietary nitrogen formulations to help fish coping with predictable stressful periods. Both metabolomic and genomic evidence show that stressful husbandry conditions affect AA metabolism in fish and may bring an increase in the requirement of indispensable AA. Supplementation in arginine and leucine, but also eventually in lysine, methionine, threonine and glutamine, may have an important role in enhancing the innate immune system. Tryptophan, as precursor for serotonin, modulates aggressive behaviour and feed intake in fish. Bioactive peptides may bring important advances in immunocompetence, disease control and other aspects of welfare of cultured fish. Fishmeal replacement may reduce immune competence, and the full nutritional potential of plant-protein ingredients is attained only after the removal or inactivation of some antinutritional factors. This review shows that AA metabolism is affected when fish are under stress, and this together with sub-optimal dietary nitrogen formulations may affect fish welfare. Furthermore, improved dietary nitrogen formulations may help fish coping with predictable stressful events.
- Different environmental temperatures affect amino acid metabolism in the eurytherm teleost Senegalese sole (Solea senegalensis Kaup, 1858) as indicated by changes in plasma metabolitesPublication . Costas, B.; Aragão, C.; Ruiz-Jarabo, I.; Vargas-Chacoff, L.; Arjona, F. J.; Mancera, J. M.; Dinis, Maria Teresa; Conceição, L. E. C.Senegalese sole (Solea senegalensis) is a eurytherm teleost that under natural conditions can be exposed to annual water temperature fluctuations between 12 and 26C. This study assessed the effects of temperature on sole metabolic status, in particular in what concerns plasma free amino acid changes during thermal acclimation.
- Effect of variable levels of dietary cholesterol and plant sterols on the growth performance and bone metabolism in gilthead seabream (Sparus aurata) juvenilesPublication . Dias, J.; Colen, Rita; Rodrigues, V.; Aragão, C.; Engrola, S.; Viegas, Michael; Laizé, Vincent; Gavaia, Paulo J.; Cancela, LeonorCholesterol is found in all animal tissues and is an important component of biological cell membranes with functions such as precursor to bile acids, hormones and vitamins. Fish meal and fish oil are cholesterol-rich ingredients. Replacement of these marine-derived ingredients by plant proteins and vegetable oils tends to reduce dietary cholesterol levels.
- Physiological short-term response to sudden salinity change in the Senegalese sole (Solea senegalensis)Publication . Herrera, M.; Aragão, C.; Hachero-Cruzado, Ismael; Ruiz-Jarabo, I.; Vargas-Chacoff, L.; Mancera, J. M.; Conceição, L. E. C.The physiological responses of Senegalese sole to a sudden salinity change were investigated. The fish were first acclimated to an initial salinity of 37.5 ppt for 4 h. Then, one group was subjected to increased salinity (55 ppt) while another group was subjected to decreased salinity (5 ppt). The third group (control group) remained at 37.5 ppt. We measured the oxygen consumption rate, osmoregulatory (plasma osmolality, gill and kidney Na+,K+-ATPase activities) and stress (plasma cortisol and metabolites) parameters 0.5 and 3 h after transfer. Oxygen consumption at both salinities was higher than for the control at both sampling times. Gill Na+,K+-ATPase activity was significantly higher for the 55 ppt salinity at 0.5 h. Plasma osmolality decreased in the fish exposed to 5 ppt at the two sampling times but no changes were detected for high salinities. Plasma cortisol levels significantly increased at both salinities, although these values declined in the low-salinity group 3 h after transfer. Plasma glucose at 5 ppt salinity did not vary significantly at 0.5 h but decreased at 3 h, while lactate increased for both treatments at the first sampling time and returned to the control levels at 3 h. Overall, the physiological response of S. senegalensis was immediate and involved a rise in oxygen consumption and plasma cortisol values as well as greater metabolite mobilization at both salinities.
- Efficiency of 2-phenoxyethanol and Clove Oil for Reducing Handling Stress in Reared Meagre, Argyrosomus regius (Pisces: Sciaenidae)Publication . Barata, Marisa; Soares, Florbela; Almeida, Ana Catarina; Pousão-Ferreira, Pedro; Ribeiro, Laura; Aragão, CláudiaThe effect of two anesthetics, 2-phenoxyethanol (2-PE) and clove oil (COil) were studied in meagre, Argyrosomus regius. This study aimed to determine the adequate dose of anesthesia for different aquaculture procedures, to assess a sedative or stressor effect of low anesthetic concentrations, and to assess the effect of low anesthetic doses at high fish densities for longer periods of time on fish survival and behavior. Anesthetics were tested at different concentrations: 2-PE at 100, 250, 400, 550, and 700mg/L; COil at 25, 40, 55, 70, and 85mg/L. Meagre became anesthetized within 10min when using concentrations above 250mg/L for 2-PE and above 40mg/L for COil. Deep anesthesia was achieved at 700 and 85mg/L, for 2-PE and COil, respectively. The fish did not react when blood was collected at these concentrations. Plasma cortisol and glucose levels were similar between sedated meagre with both 2-PE-100mg/L and COil-10mg/L, and the control (not sedated; P>0.05). This suggests that low concentrations of these anesthetics induce similar stress response as handling without anesthesia during routine activities. No mortality was registered when meagre was maintained at high densities with low concentrations of each anesthetic for 2h.
- Physiological responses of Senegalese sole (Solea senegalensis Kaup, 1858) after stress challenge: Effects on non-specific immune parameters, plasma free amino acids and energy metabolismPublication . Costas, B.; Conceição, L. E. C.; Aragão, C.; Martos-Sitcha, J. A.; Ruiz-Jarabo, I.; Mancera, J. M.; Afonso, A.Physiological responses after an acute handling stress and their subsequent effects on innate immune parameters, plasma free amino acids (AA) and liver energy substrates were assessed in Senegalese sole (Solea senegalensis). Eight groups of six specimens (136.1± 58.4 g wet weight) were maintained undisturbed, while other eight groups of six specimens were used for acute stress challenge (air exposed during 3 min). A group of six specimens was sampled for blood and head-kidney collection immediately after air exposure (time 0), while the remaining groups were sampled at 5 and 30 min, 1, 2, 4, 6 and 24 h. Undisturbed fish were sampled at the same times and used as control. Fish were fasted for 24 h prior to air exposure and sampling. Plasma cortisol, glucose, lactate and osmolality levels increased immediately after stress peaking at 1 h in air exposed fish. Changes in plasma free AA were also observed at 1 and 24 h after stress. In liver, glycogen levels significantly decreased at 30 min and 1 h, while triglycerides values significantly increased at 1, 2 and 4 h in air exposedfish. In addition, total AA levels in liver augmented significantly at 2 h holding high until 24 h in air exposed specimens. The respiratory burst of head-kidney leucocytes from air exposed fish was significantly higher than that from control groups at 2 and 6 h after air exposure. On the other hand, plasma lysozyme activity significantly decreased at 4 h after acute stress in air exposed fish, while plasma alternative complement pathway followed an inverse linear relationship with respect to cortisol showing the lowest value at 1 h after air exposure. The present study suggests that Senegalese sole presents a stress response comparable to that observed in other teleosts. While some indispensable AA may be used for the synthesis of compounds related to the stress response or fatty acid transport, dispensable AA were probably mainly employed either as energy sources or in gluconeogenesis. Moreover, results from non-specific immune parameters assessed suggest that cortisol may act as regulator of the innate immune system.
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