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Microalgae are one of the most promising groups of functional ingredients for inclusion in microdiets for Senegalese sole (Solea senegalensis) larvae, supplying components with anti-oxidant effects that may be important during an accelerated stage of development where excessive reactive oxygen substance production leading to oxidative stress may occur. The diatom Phaeodactylum tricornutum complies with such potential since it is rich in fucoxanthin, a carotenoid with demonstrated antioxidant activity. This study aimed at determining the effect of dietary P. tricornutum supplementation on the growth performance, survival and levels of oxidative stress of Senegalese sole postlarvae. Four dietary treatments were tested: a commercial microdiet, used as control, and three experimental variants where P. tricornutum was included (1) whole cells at 5%-WP5, (2) broken cells at 5%-BP5 and (3) broken cells at 10%-BP10. At the end of the trial, no significant differences were found in growth performance of larvae fed diets where P. tricornutum was included at 5% (WP5 and BP5) and the commercial control. However, larvae from the BP10 treatment had a significantly higher dry weight than larvae from the remaining treatments. Moreover, the experimental diets had no significant effects on oxidative stress. Nonetheless, lipid peroxidation levels were significantly higher in smaller larvae fed diets where P. tricornutum whole cells were included, relatively to the commercial control. The results of the present study demonstrate that P. tricornutum broken cell biomass has the potential for inclusion in weaning microdiets for Senegalese sole, increasing larval growth performance at an inclusion level of 10 %. European Union European Commission [613588] info:eu-repo/semantics/publishedVersion

Senegalese sole (Solea senegalensis) production presents several nutritional challenges, making this species a good candidate to study the dietary potential of bioactive compounds. Since proper nutrition plays a fundamental role in fish biology, it is necessary to further investigate species-specific and well-balanced diets in order to improve Senegalese sole juvenile farming. Algae have antioxidant properties, high-quality dietary protein, and are a source of bioactive compounds. This study evaluates the effects of dietary microalgal inclusion in both health status and growth performance of Senegalese sole post-larvae. Individuals 41 days after hatching (DAH) were randomly distributed among 12 tanks and four experimental diets were randomly distributed by triplicate groups. A basal diet served as CTRL and the experimental diets were formulated to include 3% of each of the algal biomass (CHLO, Chlorella sp. from heterotrophic production; PHAEO, Phaeodactylum sp.; and NANNO, Nannochloropsis sp.). At 50 DAH, 20 post-larvae/tank were collected and homogenized for analysis of immune and oxidative status, and at 61 DAH the total length, dry weight, and survival were assessed. No changes were observed in survival and total length of individuals, post-larvae fed NANNO, and CHLO dietary treatments increased dry weight at 61 DAH compared with those fed CTRL. While post-larvae immune status was apparently not altered by dietary treatments at 50 DAH, the total glutathione content decreased in fish fed PHAEO and CHLO dietary treatments compared to control diet. The observed results on improvement of growth performance without adverse effects on the immune status and decrease of endogenous total glutathione point to the fact that Nannochloropsis sp., Phaeodactylum sp., and Chlorella sp. could work as potential candidates for inclusion in microdiets for Senegalese sole. VALORMAR project - Compete 2020 [POCI-01-0247-FEDER-024517]; Lisboa 2020; CRESC Algarve 2020; Portugal 2020; European Union through European Regional Development Fund; Foundation for Science and Technology (FCT)Portuguese Foundation for Science and TechnologyEuropean Commission [UIDB/04423/2020, UIDP/04423/2020]; FCT, PortugalPortuguese Foundation for Science and TechnologyEuropean Commission [SFRH/BD/108243/2015, PD/BDE/129262/2017, IF/00197/2015] info:eu-repo/semantics/publishedVersion

Dietary phospholipids' (PLs) content, origin, and profile are known to affect fish development and reproductive performance, but their effects in zebrafish (Danio rerio) nutrition are still poorly investigated. Therefore, this study aimed to assess the effect of practical microdiets containing plant-based and marine PL sources in zebrafish growth, survival, skeletal development, and reproductive performance. Reproductive performance was evaluated according to sperm motility, number of eggs, egg morphometry, hatching rate, and offspring standard length at 5 days postfertilization (dpf). For this purpose, seven microdiets were used, where two control diets were tested along with a supplementation with soybean lecithin (SL) as a plant-based PL source, and krill oil (KO) and copepod oil (CO) as marine PL sources, or in combinations (SLCO and SLKO). KO supplementation decreased larval growth performance and induced severe skeletal anomalies. SL supplementation reduced sperm total motility but improved offspring length at 5 dpf. CO supplementation increased sperm motility and the number of spawned eggs. Our results showed that a careful selection of the origin of dietary PL sources for microdiet formulation is critical to ensure adequate skeletal development and reproductive success. This study contributes to the improvement of zebrafish microdiet formulation and optimization of zebrafish husbandry practices. FEDER through PO Algarve 21 [23000-ZEBRAFEEDS] Portuguese Foundation for Science and Technology (FCT)Portuguese Foundation for Science and Technology [SFRH/BD/97466/2013] Foundation for Science and Technology (FCT)Portuguese Foundation for Science and Technology [UID/Multi/04326/2019] CRESC Algarve 2020 [EMBRC.PT ALG-01-0145-FEDER-022121] COMPETE 2020 [EMBRC.PT ALG-01-0145-FEDER-022121] info:eu-repo/semantics/publishedVersion

Larvae and early juveniles of fast-growing fish species show tremendous growth potential, leading to higher requirements for protein, high-unsaturated fatty acids, and other nutrients. Several commercial weaning diets with relatively high success have been developed for low/moderate-growing species; however, additional challenges are outlined to meet growth potential and energy requirements of such fast-growing species. The objective of the present study was to evaluate two commercial microdiets for marine fish, one having simultaneously a very high protein and high lipid contents (HIGH), and another (MOD) with a high protein and moderate lipid content, in a growth performance trial with greater amberjack post-larvae from 33 to 78 days after hatching. Moreover, histological preparations of liver, anterior and posterior intestine were assessed for hepatic and intestinal lipid inclusions quantification and gut epithelial brush height measurement. Activities of the digestive enzymes: pepsin, trypsin, chymotrypsin, lipase and amylase were also analyzed. Post-larvae fed HIGH microdiet exhibited higher final weight and lower feed conversion ratio that those fed on MOD microdiet. Liver displayed a higher level of lipid inclusions for the MOD diet than for HIGH diet. Moreover, enterocytes of posterior intestine presented a much higher level of supranuclear vacuoles for the HIGH diet compared to MOD diet. The lower Trypsin/Chymotrypsin ratio observed at the end of the experiment in larvae fed on MOD diet may indicate a deficiency in protein of this diet. Together, these results support that larvae of greater amberjack, and likely other fast-growing marine fish species, require high protein–high lipid microdiets. The use of microdiets developed targeting slower growing marine species may lead to sub-optimal performances in fast-growing larvae., This study was undertaken by Project WEANTUNA supported by Eurostars (E! 7994) and co-financed by FEDER through PO Algarve 21 in the framework of QREN 2007-2013 (30200).

Zebrafish is a model species with a high variability of feeding regimes among fish facilities. The use of live feeds for early life stages is a common practice, and few studies have focused early weaning into microdiets. The lack of standardized feeding protocols among research facilities promotes discrepancies in biological performances, and few studies relate dietary regimes to zebrafish development. The objective of this work was to assess the effect of an early transition into microdiets in zebrafish development by evaluating growth, survival, reproductive performance, and skeletal anomalies. These parameters were assessed in one group exclusively fed on Artemia nauplii and two groups fed on microdiets (commercial and experimental). Results showed that an early weaning with the two microdiets significantly improved zebrafish growth and reproductive performance, while a decrease in incidence of vertebral column anomalies was observed. A high survival was also maintained in fish fed microdiets at an early developmental stage when comparing to exclusive Artemia nauplii feeding. In conclusion, early weaning with high quality microdiets is beneficial for zebrafish growth, reproductive performance, and skeletal development, contributing to the standardization of zebrafish husbandry practices. FEDER through PO Algarve 21 in the framework of QREN - 3000-ZEBRAFEEDS. Portuguese Foundation for Science and Technology -SFRH/BD/974662013. info:eu-repo/semantics/publishedVersion

Broodstock 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. FP7/SME/2008/1; UID/Multi/04326/2019; DL 57/2016/CP1361/CT0007; DL 57/2016/CP1361/CT0033 info:eu-repo/semantics/publishedVersion

In diet formulation for fish, it is critical to assure that all the indispensable amino acids (IAA) are available in the right quantities and ratios. This will allow minimizing dietary AA imbalances that will result in unavoidable AA losses for energy dissipation rather than for protein synthesis and growth. The supplementation with crystalline amino acids (CAA) is a possible solution to correct the dietary amino acid (AA) profile that has shown positive results for larvae of some fish species. This study tested the effect of supplementing a practical microdiet with encapsulated CAA as to balance the dietary IAA profile and to improve the capacity of Senegalese sole larvae to utilize AA and maximize growth potential. Larvae were reared at 19 A degrees C under a co-feeding regime from mouth opening. Two microdiets were formulated and processed as to have as much as possible the same ingredients and proximate composition. The control diet (CTRL) formulation was based on commonly used protein sources. A balanced diet (BAL) was formulated as to meet the ideal IAA profile defined for Senegalese sole: the dietary AA profile was corrected by replacing 4 % of encapsulated protein hydrolysate by CAA. The in vivo method of controlled tube-feeding was used to assess the effect on the larvae capacity to utilize protein, during key developmental stages. Growth was monitored until 51 DAH. The supplementation of microdiets with CAA in order to balance the dietary AA had a positive short-term effect on the Senegalese sole larvae capacity to retain protein. However, that did not translate into increased growth. On the contrary, larvae fed a more imbalanced (CTRL group) diet attained a better performance. Further studies are needed to ascertain whether this was due to an effect on the voluntary feed intake as a compensatory response to the dietary IAA imbalance in the CTRL diet or due to the higher content of tryptophan in the BAL diet.

Senegalese sole (Solea senegalensis) is a highly-valuable flatfish species with interest for diversification of aquaculture in Mediterranean countries. Unlike other fish species known to have a high dietary DHA requirement during the larval stage, several studies have suggested dietary DHA could play a minor role for Senegalese sole during the early developmental stages. This suggestion was recently supported by studies that indicated an ability of Senegalese sole to biosynthesise DHA from its precursors during the larval period. However, it is still uncertain if Senegalese sole post-larvae can effectively grow on low dietary DHA levels during weaning. Moreover, recent studies focusing on the juvenile phase of Senegalese sole have suggested a possible impairment of this species to efficiently digest or absorb high dietary lipid levels. Yet, if a reduction of dietary lipid levels would affect a normal development of Senegalese sole during the early stages remains to be established. Therefore, this study aimed at determining if a reduction of dietary DHA and lipid levels would affect Senegalese sole performance and lipidme-tabolism duringweaning and post-larval period. In this study, Senegalese sole post-larvae were reared according to four dietary treatments: low lipid levels (treatments LFAT and LFAT_DHA) and moderate lipid levels (MOD-FAT and MODFAT_DHA). DHA levels in LFAT_DHA and MODFAT_DHA diets were higher than in LFAT and MOD-FAT diets, respectively. Subsequently, effects of experimental diets on Senegalese sole post-larval growth, survival, fatty acid composition and histopathological condition of intestinal and liver tissues were evaluated. Results showed that feeding on diets with low DHA and lipid levels significantly reduced the growth performance of Senegalese sole post-larvae. However, at the end of the experimental period, no significant differences were observed for post-larval survival in response to dietary treatments. In overall, this study disadvises that weaning diets for Senegalese sole post-larvae should contain low DHA and lipid levels, as a reduction of these levels may compromise post-larval growth performance.Statement of relevance: Determining essential fatty acid requirements in marine fish larvae is of utmost importance for the aquaculture industry, contributing to fulfil the growth potential of farmed species. In the case of Senegalese sole, a highly valued species farmed in Mediterranean countries, it may have the capacity to biosynthesise DHA from its precursors during the larval stage. However, the inclusion of DHA-rich oils in weaning diets is costly, considerable savings could be achieved by replacing DHA-rich oils by common fish oils in weaning diets for this species. Furthermore, Senegalese sole has also been shown a poor ability to deal with high dietary lipid levels during the juvenile stage. However, no studies have addressed this capacity in Senegalese sole larvae. This study comprises an evaluation on the effects of reducing dietary DHA and lipid levels during Senegalese sole weaning by integrating an assessment of these effects on post-larval growth, survival, fatty acid composition and histopathological condition. (C) 2016 Elsevier B.V. All rights reserved. Project SOLEAWIN (310305/FEP/71)

23 páginas, 3 figuras, 4 tablas., 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., This work was supported by project POCI/CVT/60176/2004—SULFAAR, financed by POCI 2010 (FCT, Portugal), which is co-financed by FEDER. Wilson Pinto and Cláudia Aragão benefited from grants SFRH/BD/27889/2006 and SFRH/BPD/37197/2007 (FCT, Portugal), respectively. Manuel Yúfera contribution was funded by Consejería Innovación, Ciencia y Empresa—Junta de Andalucía (Spain) which is co-financed by FEDER (project P06-AGR-01697). This publication benefits from participation in LARVANET COST action FA0801.

Mestrado em Engenharia Física The main objective of this work is the development of a hardmetal components (WC-6%Co) recovery method by thermal deposition process. The thermal deposition technique used was HVOF (high velocity oxygen-fuel). The HVOF enables depositions of thick coatings (100-500 µm) with low porosity levels, high hardness and excellent adhesion. Before deposition, hardmetal samples with different geometries (plates and cylinders) were finished in order to have different roughness. The influence of these parameters in adhesion was studied. After this step, different re-sintering temperatures were used, in order to determine which one allows to obtain the maxima densification, elements distribution and metallurgical bonding. The re-sintering promotes the densification of the coating, with an increase of its hardness and metallurgical bonding formation. The inclusion of an intermetallic layer was tested along with different layer parameters. In liquid phase sintering (1383 and 1455 ºC) a complete densification of the coating occurred, while a bonding between the substrate and the coating only partially happened. The results of SEM/EDS show low levels of porosity and a complete and uniform distribution of the elements of the alloy. The cylindrical samples without intermetallic layer showed the lowest level of porosity and best metallurgical bonding. When the substrate surface was polished (Ra = 0.05 mm) lower levels of porosity and greater metallurgical bonding were found for both geometries. Taking into account the results obtained in this study, we can conclude that the implementation of this process is appropriate for cylindrical components with a polished surface. In these components the intermetallic layer is unnecessary and punctual defects like pores can be repaired with this process. O presente trabalho teve como objetivo o desenvolvimento de um processo de recuperação de componentes de metal duro (WC-6%Co) por projeção térmica, especificamente por HVOF (high velocity oxygenfuel). Esta técnica possibilita a obtenção de revestimentos espessos (100-500 µm) com baixos níveis de porosidade, elevada dureza e excelente adesão ao substrato. Foram testados diferentes tipos de acabamentos superficiais de modo a avaliar qual a superfície onde se obtém melhor adesão do filme e diferentes temperaturas de ressinterização, para garantir a ligação metalúrgica. A ressinterização permitiu a densificação do filme e o consequente aumento da sua dureza bem como a ligação metalúrgica do filme ao substrato. Procedeu-se também à variação da geometria dos provetes (placas e cilindros) e estudou-se ainda a variação da espessura e rugosidade de uma camada de interligação de níquel-alumínio (NiAl). Nas placas, a sinterização na presença de fase líquida (1383 e 1455 °C) deu origem a uma densificação completa do filme e apenas uma ligação metalúrgica parcial entre o revestimento e o substrato. Os resultados obtidos por SEM/EDS mostram um baixo nível de porosidade e uma difusão completa e uniforme dos elementos de liga. Nas amostras cilíndricas sem camada de interligação ocorreu uma ligação metalúrgica perfeita não sendo detetada qualquer porosidade, tendo sido observada uma elevada uniformidade no revestimento. Os melhores resultados a nível de porosidade e ligação metalúrgica foram obtidos nas superfícies polidas (Ra= 0,05 mm) em ambas as geometrias. Face aos resultados obtidos, é possível concluir que a implementação do processo desenvolvido a nível industrial, pode ser realizada em componentes cilíndricos e com superfície polida. Nesta geometria e com acabamento polido foram obtidas ligações metalúrgicas perfeitas e sem presença de porosidade, não sendo necessária a inclusão da camada de interligação de NiAl. O estudo realizado também permitiu verificar que esta técnica se poderá implementar em defeitos localizados, nomeadamente poros superficiais.

(Tesis) Ingeniero en Computación e Informática Actualmente en la empresa Anglo American División El Soldado, se presentan problemas al momento del desarrollo de los procesos de capacitación correspondiente tanto a trabajadores que deben realizarla, como a los encargados de tramitar los documentos (administración, supervisores y gerentes),por lo que como proyecto informático tenemos el objetivo reducir el tiempo que el personal de la organización ocupa en los procesos de capacitación (generación, toma y resultados), a partir de una solución informática propia para la empresa con el fin de tener un producto diferenciador de otro que se pueda implementar y que cumpla con las expectativas del cliente.

Trabajo presentado en la IV International Meeting on Marine Research (IMMR 2014), celebrada en Peniche (Portugal) el 10 y 11 de julio de 2014., The improvement of micro diets, for fish larvae, has been a major challenge in past years; however some problems remain without a resolution. Besides its tremendous growth rates, at early developmental stages, fish larvae present an immature digestive system and a progressive metamorphic process that suggest special nutritional requirements. It is therefore essential to provide fish larvae with easy and/or quickly metabolized protein sources, like protein hydrolisates, high bioavailable forms of vitamins and trace minerals. The low molecular weight of such compounds, in association to the high surface area of the micro-particles, makes them highly soluble and prone to water leaching losses. These nutrient losses, not only makes these nutrients unavailable to the larvae, but originate an important increase of organic matter in the rearing tanks, leading to detrimental water quality criteria. The development of high-performing micro-particulate diets for fish larvae requires a delicate balance between an adequate quantitative and qualitative nutritional supply and good water stability of the feeds, without compromising the palatability and the digestive breakdown. A series of binders, protein-based (e.g. fish gelatin, wheat gluten, spray-dried plasma) and carbohydrate-based (gums, alginates, pectins, starches, dextrin, polyols) were incorporated at graded levels (1, 2.5 and 5%) in a standard formulation for seabream larvae. Diets were manufactured by a proprietary low-shear extrusion process and afterwards processed to obtain a 200-400 micron fraction. All diets were analyzed for the following criteria: water leaching of nitrogenous compounds, turbidimetry and particle water stability. An additionally approach relied on the use of various encapsulation techniques (spray-drying, fluid-bed drying and vacuum coating) to incorporate trace minerals (Zn and Mn) and amino acids (taurine, lysine and methionine). Technological results show that the beneficial effects towards nutrient leaching and physical stability of the micro-pellets after immersion in water are highly dependent on binder type and less markedly on binder dose. Through the use of selected binders (e.g. guar gum and gelatin) in association to a spray-drying encapsulation process, we were able to significantly alter the release profile of Zn, Mn and taurine. Some of these prototypes were also tested in vivo with gilthead seabream larvae and clearly suggest that microencapsulation technologies that allow a controlled release profile of nutrients result in beneficial effects to larval growth and quality criteria. The results of several specific scenarios will be presented., This work has been partly funded under projects 13380-MICALA and 23000-ZEBRAFEEDS co-financed by FEDER through PO Algarve 21 in the framework of QREN 2007-2013, and by the EU seventh Framework Programme by the ARRAINA project No. 288925: Advanced Research Initiatives for Nutrition & Aquaculture.

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.

Flatfish species seem to require dietary taurine for normal growth and development. Although dietary taurine supplementation has been recommended for flatfish, little is known about the mechanisms of taurine absorption in the digestive tract of flatfish throughout ontogeny. This study described the cloning and ontogenetic expression of the taurine transporter (TauT) in the flatfish Senegalese sole (Solea senegalensis). Results showed a high similarity between TauT in Senegalese sole and other vertebrates, but a change in TauT amino acid sequences indicates that taurine transport may differ between mammals and fish, reptiles or birds. Moreover, results showed that Senegalese sole metamorphosis is an important developmental trigger to promote taurine transport in larvae, especially in muscle tissues, which may be important for larval growth. Results also indicated that the capacity to uptake dietary taurine in the digestive tract is already established in larvae at the onset of metamorphosis. In Senegalese sole juveniles, TauT expression was highest in brain, heart and eye. These are organs where taurine is usually found in high concentrations and is believed to play important biological roles. In the digestive tract of juveniles, TauT was more expressed in stomach and hindgut, indicating that dietary taurine is quickly absorbed when digestion begins and taurine endogenously used for bile salt conjugation may be recycled at the posterior end of the digestive tract. Therefore, these results suggest an enterohepatic recycling pathway for taurine in Senegalese sole, a process that may be important for maintenance of the taurine body levels in flatfish species. Electronic supplementary material The online version of this article (doi:10.1007/s00726-010-0826-6) contains supplementary material, which is available to authorized users.

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.

Aromatic amino acids (AAs, phenylalanine and tyrosine) may be specifically required during fish metamorphosis, since they are the precursors of thyroid hormones which regulate this process. This project attempted to evaluate aromatic AA metabolism during the ontogenesis of fish species with a marked (Senegalese sole; Solea senegalensis) and a less accentuated metamorphosis (gilthead seabream; Sparus aurata). Fish were tube-fed with three L-[U-14C] AA solutions at pre-metamorphic, metamorphic and post-metamorphic stages of development: controlled AA mixture (Mix), phenylalanine (Phe) and tyrosine (Tyr). Results showed a preferential aromatic AA retention during the metamorphosis of Senegalese sole, rather than in gilthead seabream. Senegalese sole's highly accentuated metamorphosis seems to increase aromatic AA physiological requirements, possibly for thyroid hormone production. Thus, Senegalese sole seems to be especially susceptible to dietary aromatic AA deficiencies during the metamorphosis period, and these findings may be important for physiologists, fish nutritionists and the flatfish aquaculture industry.

Hypersaline habitats in Chile, from marine solar salt pans to saline lagoons and pools in Andean salars, were prospected in search of Artemia populations. These saline ecosystems were characterized through physico-chemical parameters and ionic composition of their brines. Biometrics of cysts and nauplii, as well as morphometrics by using multivariate discriminant analysis for adult specimens evidenced that the Chilean populations of brine shrimp belong to A. franciscana. Cross-breeding results supported the former hypothesis of conspecific Chilean populations, and their differences with A. persimilis, also endemic to the New World, but restricted to Argentinean sites.