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Gilthead seabream (Sparus aurata) tend to increase fat deposition during summer farming conditions in the Mediterranean, which may negatively affect productive performance and consumers' quality perception of the final product. Therefore, this study evaluated the impacts of protein to lipid ratios in low fishmeal/fish oil diets on growth performance, body composition, feed conversion and nutrient utilization of seabream on-grown during summer temperature conditions. The experimental diets contained low levels of fishmeal, fish oil, and crude protein (39%), differing in crude lipid content: 16% (MF diet) or 12% (LF diet). A growth trial was per-formed with seabream (initial weight: 100 & PLUSMN; 7 g) from August to October (water temperature: 23.1 & PLUSMN; 2.2 & DEG;C). A digestibility trial was also performed (at 23 & DEG;C). Key performance indicators, whole-body composition and ac-tivities of digestive enzymes were evaluated at the end of the experiment (64 days). Low dietary lipid levels negatively affected lipid, energy, and amino acid digestibility, and as a result, fish fed the LF diet presented higher nitrogen faecal losses. Still, the decrease in nutrient digestibility was not related to dietary effects on the digestive enzyme activities. The experimental diets did not compromise the activity of pancreatic, gastric, and intestinal digestive enzymes nor feed utilization, but a slight growth impairment was observed in fish fed the LF diet, probably due to the lower amino acid and lipid digestibility. However, a potential benefit of this dietary treatment towards reducing fat accumulation in seabream during summer was observed. Nevertheless, the environmental impact of the nitrogen losses during seabream on-growing should be considered when estimating the sustainability of the production. This study demonstrated that the optimisation of diet formulations should account for the environmental conditions, especially in Mediterranean aquaculture, so the economic and envi-ronmental impacts may be correctly evaluated towards a more sustainable fish production. LA/P/0101/2020 info:eu-repo/semantics/publishedVersion

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Background: Several miRNAs are known to control myogenesis in vertebrates. Some of them are specifically expressed in muscle while others have a broader tissue expression but are still involved in establishing the muscle phenotype. In teleosts, water temperature markedly affects embryonic development and larval growth. It has been previously shown that higher embryonic temperatures promoted faster development and increased size of Senegalese sole (Solea senegalensis) larvae relatively to a lower temperature. The role of miRNAs in thermal-plasticity of growth is hitherto unknown. Hence, we have used high-throughput SOLiD sequencing to determine potential changes in the miRNA transcriptome in Senegalese sole embryos that were incubated at 15 ºC or 21 ºC until hatching and then reared at a common temperature of 21 ºC. Results: We have identified 320 conserved miRNAs in Senegalese sole, of which 48 had not been previously described in teleosts. mir-17a-5p, mir-26a, mir-130c, mir-206-3p, mir-181a-5p, mir-181a-3p and mir-199a-5p expression levels were further validated by RT- qPCR. The majority of miRNAs were dynamically expressed during early development, with peaks of expression at pre-metamorphosis or metamorphosis. Also, a higher incubation temperature (21 ºC) was associated with expression of some miRNAs positively related with growth (e.g., miR-17a, miR-181-5p and miR-206) during segmentation and at hatching. Target prediction revealed that these miRNAs may regulate myogenesis through MAPK and mTOR pathways. Expression of miRNAs involved in lipid metabolism and energy production (e.g., miR-122) also differed between temperatures. A miRNA that can potentially target calpain (miR-181-3p), and therefore negatively regulate myogenesis, was preferentially expressed during segmentation at 15 ºC compared to 21 ºC. Conclusions: Temperature has a strong influence on expression of miRNAs during embryonic and larval development in fish. Higher expression levels of miR-17a, miR-181-5p and miR-206-3p and down-regulation of miR-181a-3p at 21 ºC may promote myogenesis and are in agreement with previous studies in Senegalese sole, which reported enhanced growth at higher embryonic temperatures compared to 15 ºC. Moreover, miRNAs involved in lipid metabolism and energy production may also contribute to increased larval growth at 21 ºC compared to 15 ºC. Taken together, our data indicate that miRNAs may play a role in temperature-induced phenotypic plasticity of growth in teleosts.

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The present work examined the short- and long-term effects of three rearing temperatures on protein metabolism and growth trajectories of Senegalese sole larvae using 14C-labelled Artemia protein as feed. A first feeding trial was performed on larvae reared at 15, 18 and 21 °C (at 26, 17 and 14 days post-hatching (dph), respectively) and a second trial conducted on post-larvae after transfer to the same rearing temperature (~20 °C) (49, 35 and 27 dph, in larvae initially reared at 15, 18 and 21 °C, respectively). Temperature greatly influenced larvae relative growth rate (RGR) and survival, since growth at 15 °C was severely depressed. Protein digestibility and retention was highest at 18 °C during the first trial (85.35 ± 1.16 and 86.34 ± 2.33 %, respectively). However, during the second trial, post-larvae from 15 °C had the highest feed intake and protein digestibility (3.58 ± 1.54 and 75.50 ± 1.35 %, respectively), although retention was similar between treatments. Furthermore, after transfer to 20 °C larvae from 15 °C experienced compensatory growth, which was observed until 121 dph, and confirmed by RGR values, which were significantly higher at 15 ºC than at 21 ºC or 18 ºC. Results from the present study show that Solea senegalensis larval development, survival and protein digestion and retention are highly affected by thermal history.

Fertilised eggs of Senegalese sole were incubated at 15, 18 or 21 °C, and after hatching all larvae were reared at 21 °C until 30 days post-hatch. By this point larvae from the 18 or 21 °C temperature groups had 11 and 9% more muscle fibres than those from 15 °C, respectively. Hyperplastic growth during metamorphosis was higher in larvae from 18 °C. Embryonic temperature induced gene expression changes, albeit with a variable pattern throughout development. Myf5, myod2, myHC and fst mRNA levels were significantly higher at several stages prior to hatching in embryos incubated at 21 °C, whereas hsp90AB and hsp70 transcripts were present at higher levels in the 15 °C group. Myf5, myod1, myod2, pax7, myog, fst, igf-II, igf1r, hsp90AA and hsp90AB were expressed at higher levels during early development, particularly during somitogenesis. In contrast, mrf4, myHC, mylc2, igf-I, mstn1 and hsp70 were up-regulated at later stages of larval development, namely during and after metamorphosis. This study is the first example of thermal plasticity of myogenesis with prolonged effect in a flatfish.

Enhanced production of high quality and healthy fry is a key target for a successful and competitive expansion of the aquaculture industry. Although large quantities of fish larvae are produced, survival rates are often low or highly variable and growth potential is in most cases not fully exploited, indicating significant gaps in our knowledge concerning optimal nutritional and culture conditions. Understanding the mechanisms that control early development and muscle growth are critical for the identification of time windows in development that introduce growth variation, and improve the viability and quality of juveniles. This literature review of the current state of knowledge aims to provide a framework for a better understanding of fish skeletal muscle ontogeny, and its impact on larval and juvenile quality as broadly defined. It focuses on fundamental biological knowledge relevant to larval phenotype and quality and, in particular, on the factors affecting the development of skeletal muscle. It also discusses the available methodologies to assess growth and larvae/juvenile quality, identifies gaps in knowledge and suggests future research directions. The focus is primarily on the major farmed non-salmonid fish species in Europe that include gilthead sea bream, European sea bass, turbot, Atlantic cod, Senegalese sole and Atlantic halibut.

In the present study, Senegalese sole eggs incubated at 20 °C were reared at three different temperatures (15 °C, 18 °C or 21 °C) during the pelagic phase, and then transferred to a common temperature (20 °C) from benthic stage until 100 days post-hatch (dph). Somatic growth, fast muscle cellularity and expression of 15 growth-related genes were compared at the same developmental stage for each temperature (pre-metamorphic larvae, metamorphic larvae, post-metamorphic larvae) and at 83 dph and 100 dph early juveniles. During pre-metamorphosis and metamorphosis, larvae from 21 °C weighed significantly more than those reared at 18 °C or 15 °C (P < 0.001). Relative growth rate (RGR) of pelagic larvae and survival of newly-settled larvae were also higher at 21 °C (P < 0.05). Furthermore, an increase in muscle growth towards the highest temperatures was observed concomitantly with an increase in gene expression, namely myogenic regulatory factors (MRFs), myosins, igf-I and fgf6. After transfer to a common temperature (20 °C), the 15 °C group initiated a process of compensatory growth, inverting relative growth rate values that became the highest, particularly between 83 dph and 100 dph (P < 0.05). This increased growth effort in the 15 °C group was accompanied by an up-regulation of gene expression in fast muscle, particularly in the 83 dph juveniles. Mrf4 and myHC were up-regulated at 15 °C and positive correlations with growth were also found for myog or pax7 (P < 0.05). In contrast, mstn1 was down-regulated at 15 °C (P < 0.05), suggesting a boost in muscle growth. By 100 dph, the 15 °C group had already reached the same weight as the 18 °C and the same length as the 21 °C ones. At this age, the 18 °C juveniles had the largest mean muscle fibre diameter (P < 0.001), and thus fibre hypertrophy seems to be a major growth mechanism relatively to the other groups. Our work shows that temperature during pelagic phase greatly influences the growth trajectory of Senegalese sole juveniles.

Texto integral disponível através do editor Myogenin (myog) encodes a highly conserved myogenic regulatory factor that is involved in terminal muscle differentiation. It has been shown in mammals that methylation of cytosines within the myog promoter plays a major role in regulating its transcription. In the present study, the Senegalese sole (Solea senegalensis) myog putative proximal promoter was identified and found to be highly conserved among teleosts. Therefore, it is plausible that it plays a similar role in controlling myog expression. Cytosine methylation of the myog promoter in skeletal muscle of Senegalese sole larvae undergoing metamorphosis was influenced by rearing temperature. A lower temperature (15°C) significantly increased myog promoter methylation in skeletal muscle, particularly at specific CpG sites, relatively to higher rearing temperatures (18 and 21°C). Myog transcription was downregulated at 15°C, whereas expression of dnmt1 and dnmt3b was upregulated, consistently with the higher myog methylation observed at this temperature. Rearing temperature also affected growth and fast muscle cellularity, producing larger fibers at 21°C. Taken together, our data provide the first evidence of an epigenetic mechanism that may be underlying the temperature-induced phenotypic plasticity of muscle growth in teleosts.

Once spawned and fertilized the oviparous fish eggs operate as closed systems; only respiratory gases, heat and negligible amounts of solutes and water are exchanged freely, as a result of an extremely low permeability of the egg surface membranes. Currently, the opportunities to exert a nutritional stimulus during a stage of high metabolic plasticity, such as fish embryogenesis, are restricted to maternal transfer and the onset of exogenous feeding.

Cholesterol 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.

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.

Temperature-induced responses at the level of muscle growth potential have been observed in teleost fish species, concomitantly with gene expression changes (Johnston and Hall 2004). Incubation temperature has been demonstrated to influence the muscle cellularity at hatch and during larval development, and these effects are highly species-specific. Changes in the relative timing and expression level of myogenic genes were found to be related with thermal plasticity of muscle phenotype (Fernandes, Mackenzie et al. 2006). The eggs of the flatfish Senegalese sole (Solea senegalensis) in aquaculture conditions are normally incubated within the 16-20ºC range, but influence of temperature on muscle cellularity and expression of growth-related genes is unknown. Understanding these effects may contribute to improve growth conditions and juvenile production.

Senegalese sole is distributed along the Mediterranean and southwestern Europe, reaching a market price of 20-25 €/kg - Promising species for aquaculture diversification - Control and intensification of culture is reasonably achieved - The high incidence of skeletal anomalies in intensively reared fish represents a constraint to commercial production - Samples from Portugal and Greece were analyzed towards identifying skeletal differences due to rearing conditions

A large effort has been dedicated in the past years to the development of nutritional balanced inert diets for marine fish larvae in order to suppress the nutritional deficiencies of live feed. In this study growth performance, Artemia intake, protein digestibility and protein retention were measured for Senegalese sole (Solea senegalensis Kaup), in order to provide insight into how protein utilization affects growth performance. Three feeding regimes were tested: ST – standard live feed; ArtRL – live feed and 20%Artemia replacement with inert diet (dry matter basis) from mouth opening; ArtRH – live feed and 58%Artemia replacement with inert diet from mouth opening. Artemia intake and protein metabolism were determined at 6, 15 and 21 days after hatching using 14C-labelled Artemia protein and subsequent incubation in metabolic chambers. At the end of the experiment, sole fed exclusively with live feed were significantly larger than sole from Artemia replacement treatments. Protein digestibility decreased during sole ontogeny, and more sharply in ArtRH sole. Concomitantly retention efficiency increased during ontogeny but with a slight delay in ArtRH sole. Senegalese sole larvae growth and protein utilization is depressed when co-fed high levels of inert diet and Artemia, mostly during metamorphosis climax.

The present study intended to evaluate the effects of early introduction of inert diet in lipid digestibility and metabolism of sole, while larval feed intake, growth and survival were also monitored. Solea senegalensis larvae were reared on a standard live feed regime (ST) and co-feeding regime with inert diet (Art R). Trials using sole larvae fed with Artemia enriched with two different lipid emulsions, containing glycerol tri [1-14C] oleate (TAG) and L-3-phosphatidylcholine-1,2-di-[1-14C] oleoyl (PL), were performed at 9 and 17 days after hatching (DAH) to study lipid utilization. Co-feeding did not affect sole survival rates (ST 59.1 ± 15.9%; Art R 69.56 ± 9.3%), but was reflected in significantly smaller final weight at 16 DAH (ST 0.71 ± 0.20; Art R 0.48 ± 0.14 mg). Higher feed intake was observed in sole larvae fed on Artemia enriched with labeled PL at 9 DAH but not at 17 DAH. At 17 DAH, the smaller larvae (Art R treatment) ingested proportionally more Artemia in weight percentage, independently of enrichment. At 9 DAH lipid digestibility was equal among treatments and higher than 90%, while at 17 DAH it was higher in ST treatment (around 73%) compared to the Art R group (around 66%). Lipid retention efficiency at 9 DAH was higher in the Art R treatment, reaching values of 50%, while these values almost duplicated at 17 DAH, ranging up to 80% in both treatments without significant differences. These results show that co-feeding of live feed and inert diet from first-feeding in Senegalese sole has a toll in terms of growth and lipid digestibility but does not seem to compromise lipid metabolic utilization.

One of the major obstacles to propagating dusky grouper, Epinephelus marginatus, is the difficulty of rearing the early larvae. We have successfully raised dusky grouper larvae in mesocosms using a mixed diet of endogenous plankton grown in the rearing tank and an exogenous supply of Brachionus plicatilis and Artemia sp. Newly hatched larvae at an initial density of 1.3 ind.l-1 were stocked in partially shaded 3 m3 circular outdoor tanks during the summers of 2007 and 2008. Before introducing newly hatched larvae, the water was left for six days to promote plankton growth. Larval growth occurred at two different rates: i) a faster rate from first feeding to the beginning of metamorphosis, and ii) a slower rate at transformation. Survival at the beginning of metamorphosis was less than 10% (33 DPH) in 2007 and between 25 and 50% (25 DPH) in 2008. High mortalities were observed during larval transformation. The estimated minimum food requirement per grouper larvae increased more than 300% from the beginning of the notochord flexion to the beginning of metamorphosis. To meet such a high feeding requirement the number of larger prey organisms/copepods in the mesocosm should be eight times higher during this time period.

Submitted by Paulo Gavaia (pgavaia@ualg.pt) on 2014-05-30T10:05:46Z No. of bitstreams: 1 Avanços recentes em nutrição de larvas de peixes.pdf: 75116 bytes, checksum: 728a03d131b35c9646da86008536f71d (MD5) Approved for entry into archive by Pedro Sousa (a19661@ualg.pt) on 2014-06-25T09:13:19Z (GMT) No. of bitstreams: 2 Avanços recentes em nutrição de larvas de peixes.pdf: 75116 bytes, checksum: 728a03d131b35c9646da86008536f71d (MD5) 8753041680192686.zip: 69092 bytes, checksum: 482c5f4899029bb21c5d8512b89342fe (MD5) Made available in DSpace on 2014-06-25T09:13:19Z (GMT). No. of bitstreams: 2 Avanços recentes em nutrição de larvas de peixes.pdf: 75116 bytes, checksum: 728a03d131b35c9646da86008536f71d (MD5) 8753041680192686.zip: 69092 bytes, checksum: 482c5f4899029bb21c5d8512b89342fe (MD5) Previous issue date: 2009

The appearance of skeletal deformities is a major problem associated to intensive, large-scale culture of fish species, affecting growth, development and ultimately, survival of produced fish. Although the mechanisms working behind it are still unclear, it is known that environmental, genetic and nutritional factors play fundamental roles. Given its importance as a vertebrate model to study skeleton-related diseases, the selection of an appropriate dietary protocol is a critical step in order to diminish the nutritional effect on bone and cartilage development. To better understand the effect of diet on skeletal development and deformities, as well as on larvae and juvenile global performance, we have tested four distinct dietary regimes, composed of rotifers Brachionus sp., Artemia nauplii and dry, formulated food.

Tese dout., Aquacultura, Universidade do Algarve, 2008 Made available in DSpace on 2011-09-07T16:05:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2008

Submitted by Paulo Gavaia (pgavaia@ualg.pt) on 2014-05-30T10:27:14Z No. of bitstreams: 1 Effect of pre-weaning feeding regime on weaning performance of Senegalese sole, Solea senegalensis (Kaup, 1858).pdf: 328434 bytes, checksum: aee6e779dae7e86c9ed402c159644123 (MD5) Approved for entry into archive by Pedro Sousa (a19661@ualg.pt) on 2014-06-25T10:35:29Z (GMT) No. of bitstreams: 2 Effect of pre-weaning feeding regime on weaning performance of Senegalese sole, Solea senegalensis (Kaup, 1858).pdf: 328434 bytes, checksum: aee6e779dae7e86c9ed402c159644123 (MD5) 8753041680192686.zip: 295206 bytes, checksum: 7a92aa469c468cff66f427c2e20c9800 (MD5) Made available in DSpace on 2014-06-25T10:35:29Z (GMT). No. of bitstreams: 2 Effect of pre-weaning feeding regime on weaning performance of Senegalese sole, Solea senegalensis (Kaup, 1858).pdf: 328434 bytes, checksum: aee6e779dae7e86c9ed402c159644123 (MD5) 8753041680192686.zip: 295206 bytes, checksum: 7a92aa469c468cff66f427c2e20c9800 (MD5) Previous issue date: 2005

The Senegalese sole, Solea senegalensis Kaup, 1858, has a high market value, and is therefore considered a potentially valuable new species for aquaculture. To attain its potential, it is necessary to improve the production of juveniles, so that production of market-size fish can get fully underway. The mass production of sole in aquaculture will only be possible when hatcheries begin to be able to produce juveniles for ongrowing. Several studies have been conducted to improve sole production (Dinis, 1986, 1992; Dinis et al., 1999), and some pigmentation abnormalities were reported, which require further study. The present paper describes the principal abnormalities found in sole pigmentation. Studies indicate that 11 % of juveniles present pigmentation problems, which could be considered a problem insofar as mass production is involved, because such fish would be difficult to market. El lenguado Solea senegalensis Kaup, 1858 es una especie de valor comercial elevado y está considerada de interés potencial para la acuicultura. En consecuencia, existe una creciente necesidad de desarrollar el cultivo larval de esta especie a escala industrial y producir juveniles para engorde en las piscifactorías. Se han realizado diversos trabajos con el objetivo de mejorar la metodología en la producción de S. senegalensis (Dinis, 1986, 1992; Dinis et al., 1999) y se señalan algunas limitaciones: entre ellas, las anomalías de pigmentación. Se describen aquí las principales anomalías en la pigmentación del lenguado a partir de puestas naturales de reproductores salvajes mantenidos en cautividad. El porcentaje total de alevines mal pigmentados fue del 11 %, lo que puede constituir un problema cuando se pretende la producción a gran escala, pues entraña un obstáculo para su comercialización, con el consiguiente descenso de la rentabilidad. Instituto Español de Oceanografía

Submitted by Sofia Engrola (sengrola@ualg.pt) on 2014-07-03T14:35:24Z No. of bitstreams: 1 Hormonal induction of Atlantic dusky grouper (Epinephelus marginatus) broodstock.pdf: 639332 bytes, checksum: ff4b06b1501e0bb9a900cf9035acb146 (MD5) Approved for entry into archive by Merja Muzavor (mmuzavor@ualg.pt) on 2014-07-04T12:33:16Z (GMT) No. of bitstreams: 2 8854940252609639.zip: 627579 bytes, checksum: 29f7949ef4356f4ed834acdafd3458b5 (MD5) Hormonal induction of Atlantic dusky grouper (Epinephelus marginatus) broodstock.pdf: 639332 bytes, checksum: ff4b06b1501e0bb9a900cf9035acb146 (MD5) Made available in DSpace on 2014-07-04T12:33:16Z (GMT). No. of bitstreams: 2 8854940252609639.zip: 627579 bytes, checksum: 29f7949ef4356f4ed834acdafd3458b5 (MD5) Hormonal induction of Atlantic dusky grouper (Epinephelus marginatus) broodstock.pdf: 639332 bytes, checksum: ff4b06b1501e0bb9a900cf9035acb146 (MD5) Previous issue date: 2008

Dissertação mest., Aquacultura e Pescas, Universidade do Algarve, 2009 The general aim of this study was to understand the underlying factors responsible for the individual variation in growth of juvenile Senegalese sole (Solea senegalensis). Senegalese sole is a species of high commercial value and high interest for aquaculture in Europe, therefore it would be important to understand these individual variations. Understanding the causes for individual variation in growth in relation with the feed intake, metabolic rate and response to stress contributes to maximize the production efficiency by reducing food waste and improving water quality. In order to identify the possible factors responsible for the individual variation in sole growth, its relationship with metabolic rate, stress response and feed intake was determined. Twelve fish with 18.49 ± 2.94g of initial weight were kept in standard conditions and feed once a day, during a period of six months. Individual oxygen consumption measurements in Sole juveniles were determined by flow-through respirometry at standard conditions, after the fish were fed a single meal and after a stressful condition. Blood samples for plasma cortisol, were taken from all fish at the start of the experiment (control, indicative of basal levels) and after stress exposure. Quantification of individual feed intake was performed by incorporation with radio-opaque ballotinis glass beads in a dry feed, followed by radiography to quantify the amount of ingested pellets. Individual fish exhibited pronounced variation in growth (final weight ranging from 30.45 to 67.53g), oxygen consumption of fasted fish (108.0 to 447.4 μmol/g/h), oxygen consumption of fed fish (174.7 μmol/g/h to 813.5 μmol/g/h), oxygen consumption after emersion stress (186.8 μmol/g/h to 376.7 μmol/g/h), and plasma cortisol after emersion stress (4.81 ng/ml to 46.45 ng/ml). Individual differences in metabolic rate do not contribute to explain individual differences in growth. However, after emersion stress individual differences in stress response seem to play a role in explaining individual differences in growth, what may result from different copying styles. Sole feed intake as measured by X-ray does not seem to be a good indicator of growth rate. The oxygen consumption increases both when fish are fed and after an emersion stress, as result of additive costs of growth and stress to the cost of maintenance. All together, this study suggests that individual variation of growth in Solea senegalensis is essentially due to genetic variation, and its reduction may accomplished when genetic selection programs are adopted. Furthermore, a study with a larger number of fish and over a larger time period would be important to check some of the trends found in this thesis between growth potential, metabolic rate and coping styles of Senegalese sole.