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Organic acids (OA) and nature-identical compounds (NIC) such as monoterpenes and aldehydes are well-known growth and health promoters in terrestrial livestock while their application for fish production is recent and their mechanisms of action require further study. Hence, this study tested the increasing dietary level (D0, D250, D500, D1000; 0, 250, 500 and 1000 mg kg feed−1 respectively) of a microencapsulated blend containing citric and sorbic acid, thymol and vanillin over 82 days on rainbow trout to assess the effects on growth, feed utilization, intestine cytokine gene expression and gut microbiota (GM). Furthermore, the effects on intestinal cytokine gene expression and GM were also explored after one week at high water temperature (23 °C). OA and NIC improved specific growth rate (SGR) and feed conversion rate (FCR) during the second half (day 40–82) of the feeding trial, while at the end of the trial protein (PER) and lipid efficiency (LER) increased with increasing dietary level. GM diversity and composition and cytokine gene expression analysis showed no significant differences in fish fed with increasing doses of OA and NIC (82 days) demonstrating the absence of inflammatory activity in the intestinal mucosa. Although there were no statistical differences, GM structure showed a tendency in clustering D0 group separately from the other dietary groups and a trend towards reduction of Streptococcus spp. was observed in the D250 and D1000 groups. After exposure to high water temperature, lower GM diversity and increased gene expression of inflammatory intestinal cytokines were observed for both inclusions (D0 vs. D1000) compared to groups in standard condition. However, the gene up-regulation involved a limited number of cytokines showing the absence of a substantial inflammation process able to compromise the functional activity of the intestine. Despite further study should be conducted to fully clarify this mechanism, cytokines up-regulation seems to be concomitant to the reduction of the GM diversity and, particularly, to the reduction of specific lactic acid bacteria such as Leuconostoc. The application of the microencapsulate blend tested can be a useful strategy to improve growth and feed utilization in rainbow trout under normal temperature conditions. According to the results organic acids and nature-identical compounds did not revert the effects triggered by the increased temperature of water.

Simple Summary Nutritional strategies focused on the use of botanicals as modulators of several physiological responses and health promoters of the gastrointestinal tract have attracted interest in animal production. Previous research indicates the positive results of using essential oils (EOs) as natural feed additives for several farmed animals. In the last decades, these nutritional alternatives have been evaluated and reported in fish production in order to increase fish growth and feed utilization and to promote animal welfare. Therefore, the present study was designed to compare the effects of feed EO supplementation in two different forms (natural and composed of active ingredients obtained by synthesis) on the gastric mucosa in European sea bass. EOs decrease oxyntopeptic cells and increase somatostatin and ghrelin enteroendocrine cells. In addition, we showed that Na+K+-ATPase was expressed in oxyntopeptic cells (OPs) in the same way as H+K+-ATPase (typical marker for mammalian parietal cells) and, for this reason, consider Na+K+-ATPase a valid marker for OPs. Abstract The current work was designed to assess the effect of feed supplemented with essential oils (EOs) on the histological features in sea bass’s gastric mucosa. Fish were fed three diets: control diet (CTR), HERBAL MIX® made with natural EOs (N-EOs), or HERBAL MIX® made with artificial EOs obtained by synthesis (S-EOs) during a 117-day feeding trial. Thereafter, the oxyntopeptic cells (OPs) and the ghrelin (GHR) and somatostatin (SOM) enteroendocrine cells (EECs) in the gastric mucosa were evaluated. The Na+K+-ATPase antibody was used to label OPs, while, for the EECs, anti-SOM and anti-GHR antibody were used. The highest density of OP immunoreactive (IR) area was in the CTR group (0.66 mm2 ± 0.1). The OP-IR area was reduced in the N-EO diet group (0.22 mm2 ± 1; CTR vs. N-EOs, p < 0.005), while in the S-EO diet group (0.39 mm2 ± 1) a trend was observed. We observed an increase of the number of SOM-IR cells in the N-EO diet (15.6 ± 4.2) compared to that in the CTR (11.8 ± 3.7) (N-EOs vs. CTR; p < 0.05), but not in the S-EOs diet. These observations will provide a basis to advance current knowledge on the anatomy and digestive physiology of this species in relation to pro-heath feeds.

In Mediterranean aquaculture, significant advances have been made towards a reduction of marine-derived ingredients in aquafeed formulation, as well as in defining the effect on how environmental factors such as rearing density interact with fish health. Little research, however, has examined the interaction between rearing density and dietary composition on main key performance indicators, physiological processes and gut bacterial community. A study was undertaken, therefore to assess growth response, digestive enzyme activity, humoral immunity on skin mucus, plasma biochemistry and gut microbiota of gilthead sea bream (Sparus aurata, L. 1758) reared at high (HD, 36–44 kg m−3) and low (LD, 12–15 kg m−3) final stocking densities and fed high (FM30/FO15, 30% fishmeal FM, 15% fish oil, FO) and low (FM10/FO3; 10% FM and 3% FO) FM and FO levels. Isonitrogenous and isolipidic extruded diets were fed to triplicate fish groups (initial weight: 96.2 g) to overfeeding over 98 days. The densities tested had no major effects on overall growth and feed efficiency of sea bream reared at high or low FM and FO dietary level. However, HD seems to reduce feed intake compared to LD mainly in fish fed FM30/FO15. Results of digestive enzyme activity indicated a comparable digestive efficiency among rearing densities and within each dietary treatment even if intestinal brush border enzymes appeared to be more influenced by stocking density compared to gastric and pancreatic enzymes. Plasma parameters related to nutritional and physiological conditions were not affected by rearing densities under both nutritional conditions a similar observation was also achieved through the study of lysozyme, protease, antiprotease and total protein determination in skin mucus, however; in this case lysozyme was slightly reduced at HD. For the first time on this species, the effect of rearing density on gut bacterial community was studied. Different response in relation to dietary treatment under HD and LD were detected. Low FM-FO diet maintained steady the biodiversity of the gut bacterial community between LD and HD conditions while fish fed high FM-FO level showed a reduced biodiversity at HD. According to the results, it seems feasible to rear gilthead sea bream at the on-growing phase at a density up to 36–44 kg m−3 with low or high FM-FO diet without negatively affecting growth, feed efficiency, welfare condition and gut bacterial community. info:eu-repo/semantics/acceptedVersion

Fishmeal (FM) possesses one of the highest buffering capacities in comparison to most alternative vegetable aquafeed ingredients and its decreasing content in current formulations might affect the ideal gastrointestinal environment for digestive enzyme action and gut bacterial community of carnivorous fish species. A study was undertaken, therefore, to assess growth response, gastrointestinal pH, digestive enzyme activity and gut bacterial community of European sea bass (Dicentrarchus labrax L.) juveniles fed low FM diets (10% FM) with or without a feed buffering additive, calcium carbonate (FM10 + B and FM10, respectively) in comparison to a standard FM diet (20% FM, FM20). Three isonitrogenous and isolipidic extruded diets were fed to triplicate fish groups of 80 individuals (initial weight: 23 g) to overfeeding over 64 days. No significant differences due to low FM dietary levels were observed in final body weight, specific growth rate, feed intake, feed and protein efficiency. Low FM diet did not affect gastrointestinal pH in the stomach, anterior intestine, mid-intestine and hindgut at 0, 4, 8 and 12 hours post meal (hpm) while the inclusion of calcium carbonate in low FM diet seems to slightly increase the pH in the hindgut at 12 hpm. The absence of significant differences of pepsin, chymotrypsin, amylase and lipase activity suggests a comparable digestive efficiency among treatments although trypsin activity was slightly reduced in low FM diets after 4 hpm. Decreasing FM content seems to exert an effect on the overall gut bacterial community analysed by next-generation sequencing even if no significant effects on specific bacterial component were detected. The gut bacterial community in all the treatments was particularly rich in lactic acid bacteria such as Lactobacillus spp. which may provide important beneficial functions for the host and be associated with a healthy intestinal epithelium. According to the results, increasing the feed buffering capacity does not seem to improve digestive conditions while it is feasible to include 10% FM dietary level in practical formulation for European sea bass juveniles without negatively affecting growth, feed efficiency and digestive luminal conditions. info:eu-repo/semantics/publishedVersion

Recent important developments in sole aquaculture have increased the availability of the farmed product on the market. The aim of this research was to characterize and compare for the first time in common sole (Solea solea) morphometric parameters, nutrients and sensory traits of cultured and wild specimens. Farmed sole, while maintaining the characteristics of a lean fish species (2% of lipid content), displayed a EPA+DHA fillet content more than twice as high as its wild counterpart. Sensory traits of “potato”, “boiled fish”, sweet, firmness, astringency, chewiness, were correlated to farmed origin while whiteness, briny, “octopus”, “crab”, salty, acid, bitter, umami, intensity and juiciness were linked to wild origin. Intensive farming conditions improved the nutritional value of common sole in terms of lipid content and fatty acid profile and led to sweeter sensory traits particularly associated with a terrestrial vegetable perception.

A 121-day feeding trial was undertaken to test the effects of two dietary lipid levels (16 and 21% L16, L21) in triplicated gilthead sea bream groups (initial weight: 67.5 g) reared at two different water temperatures (high, H 23°C and low, L 17°C) in the same recirculation system but exposed to a switch in temperature after 58 days. Fish kept at H were transferred to L (HL transition, autumn shift), and the fish kept at L were exposed to H (LH transition, summer shift), while continuing to receive the same diet to apparent satiation in each group. At the end of the trial, no significant diet effect on specific growth rate (SGR), feed intake (FI), and feed conversion rate (FCR) were detected in fish exposed to HL transition compared with those exposed to LH transition, while gross lipid efficiency (GLE) and lipid efficiency ratio (LER) were higher in L16. After temperature changes, L16 displayed higher SGR, FI, GLE, and LER, while mesenteric fat index was reduced. After temperature changes, the combined effects of low lipid diet and low temperature conditions resulted in higher pepsin activity, while trypsin, chymotrypsin, and lipase activities were generally higher at high lipid content. The combined effect of diet and temperature did not alter the metabolic plasma profile, except for the observed final higher aspartate aminotransferase (AST) and alkaline phosphatase (ALP) values when combining high dietary lipid (L21) and temperature changes. Different diets showed a significantly different gut microbiome layout, only at high temperature with L16 diet resulting in a higher load of Lactobacillus. On the contrary, no dietary impact on ecosystem diversity was observed, independently from the temperature. In addition, L16 diet in the HL transition favored an increase in Weissella and Bradyrhizobium genera in the gut microbiome, while in the final condition of LH transition, L21 diet favored a significant increase in Streptococcus and Bacillus. According to the results, the utilization of 16% dietary lipid levels in gilthead sea bream should be preferred during seasonal temperature changes in order to optimize feed utilization and gut health., This research was undertaken under the MedAID (Mediterranean Aquaculture Integrated Development) project, which has received funding from the European Union’s Horizon 2020 Research and Innovation Programme, Call H2020-SFS-23-2016, Grant agreement no. 727315 (http://www.medaid-h2020.eu/). MY and NG received support from the Spanish Ministry of Science, Innovation and Universities (MCIU), the State Research Agency (AEI), the European Regional Development Fund (FEDER/ERDF), and project Thermodigest (RTI2018-096134-B-I00) granted to MY.

A 71-day study was conducted to explore the effect of increasing dietary levels (0, 250, 500, 1000 mg kg feed−1; D0, D250, D500 and D1000, respectively) of a blend of microencapsulated organic acids (OA, specifically citric and sorbic acid) and nature identical compounds (NIC, specifically thymol and vanillin), on growth, intestinal immune parameters and gut microbiota (GM) of European sea bass juveniles reared under normal and subsequently suboptimal environmental conditions (high temperature, 30.0 ± 0.4 °C and low oxygen, 4.6 ± 0.6 mg L−1). OA and NIC did not promote growth, feed utilisation and feed intake at the inclusion tested but induced a significantly upregulation of IL-8, IL-10 and TGFβ. GM analyzed by next-generation sequencing showed that OA and NIC were able to exert prebiotic properties stimulating the development of beneficial bacteria taxa such as Lactobacillus, Leuconostoc, and Bacillus sp. Picrust analyses displayed a significant potential functional reconfiguration of GM promoting a decrease in inflammation-promoting and homeostatic functions at increasing OA and NIC administration. For the first time on this species the exposure to suboptimal rearing conditions was able to modify GM structure reducing LAB and increasing Proteobacteria, findings which were consistent with the inflammatory process observed at mRNA level.

The high nutritional quality of common sole Solea solea increases its value for the fishery industry and for the aquaculture sector. To ensure the expansion of its production, it is necessary to implement farming and broodstock management technique to produce high quality eggs and larvae. This work summarizes eight years of study on reproductive performances, growth and parental contribution of a common sole broodstock from the Mediterranean-North Adriatic Sea. The broodstock (11 females and 13 males) reached the peak of fecundity after 5/6 years of captivity, with a production of 296.48 and 376.54 × 103 of total eggs kg−1 female with a fertilization rate of 31.6 ± 18.3 and 41.9 ± 23.8 %, respectively. Results shows that variations in temperature cycles are pivotal for a successful breeding season, and body condition during the first 3 years of captivity was the most important parameter positively correlated to parental contribution for both males and females. Parental contribution was assessed by eight species-specific microsatellite loci during the first 3 years of reproduction. Although the two sexes displayed different temporal trends, parentage assessment demonstrated an overall increase in the number of active breeders. However, only 13 couples out of 50 produced the 70.4 % of the larvae in year 5, confirming the same reproduction behavioural pattern in Mediterranean common sole than that observed both in Senegalese sole and in common sole of Atlantic origin. This work provides valuable data needed to understand the mechanisms behind the maturation characteristics of this species and support future strategy for the establishment and management of Mediterranean common sole aquaculture.

In the issue of fishmeal replacement, besides maintaining optimal growth, a key area of investigation for continuing to improve modern aquafeeds includes the evaluation of the effects of plant ingredients on fish quality. It is generally accepted that farmed fish quality can be influenced by the formulation of composition of their feed. Hence, the aim of the present research was to evaluate plant protein inclusion up to 84% of the overall protein content in an integrated study on growth and quality traits of European sea bass. Three diets were formulated to contain increasing plant protein levels (50, 67 and 84%; 50PP, 67PP and 84PP, respectively), with fishmeal dietary levels at 30, 20 and 10%, respectively. No significant differences due to reducing fishmeal content were observed after 118 days in terms of growth (final body weight and specific growth rate) and feed intake, even though a trend towards lower growth performance at higher fishmeal replacement levels was observed. Fish fed diet 50PP showed lower feed conversion rate in comparison to those fed diet 84PP, while no differences were recorded between diet 50PP and 67PP. No significant differences among treatments were found in protein efficiency rate. On the contrary, fish fed diet 84PP showed lower gross protein efficiency in comparison to those fed diet 50PP and 67PP. No significant differences in biometric indices and fillet composition were observed. No significant differences were found in pH, liquid holding capacity and skin colour measurements between treatments, while regarding fillet colour, significant differences were found only for H°ab. In conclusion, our findings demonstrated that dietary plant proteins up to 84% of the overall protein content had no effects on quality traits of European sea bass in comparison with 50% and 67%. All experimental groups showed similar growth even though 84% plant protein inclusion negatively influenced feed and protein utilisation.

Contents The study of lipids and fatty acids (FAs) has been used in the assessment of egg quality because their composition can influence the fertilization rate, hatching, survival and growth of marine fish larvae. For these reasons, the lipid content (TL) and fatty acid composition of common sole (Solea solea) eggs were measured and correlated to egg and larval viability parameters throughout an entire reproductive season. Seventeen batches of fertile eggs obtained from natural spawning of captive breeders were characterized for the TL, FA profile, hatching rate (HR) and survival rate of larvae (SR) at 0–6 days post-hatching (dph). The egg FA composition reflected the composition of the feed supplied to the broodstock during summer and autumn (before and during vitellogenesis) rather than that supplied during the spawning season. In general, the egg FA profile showed minimal differences among the early-, mid- and late-spawning periods (possibly due to the change of the diet and/or water temperature) indicating that it is possible to obtain a similar egg quality in terms of egg FA profile over 2 months of spawning. Saturated FAs and monounsaturated FAs (MUFA) were positively correlated with HR, while TL, 22 : 6n-3 (DHA), 20 : 4n-6 (ARA), polyunsaturated FAs of the (n-3) series (n-3 PUFA) and polyunsaturated FAs of the (n-6) series were negatively correlated (p ≤ 0.05). MUFA, 20 : 5n-3 (EPA), n-6/n-3 were positively correlated with SR, while DHA, n-3 PUFA, DHA/EPA were negatively correlated (p ≤ 0.05). In conclusion, the feed supplied before and during vitellogenesis has a major role in determining the egg FA profile in common sole. The relationships found between TL and FAs with egg and larval viability parameters differ from many other farmed marine fish species, which may suggest the need for a specific broodstock feed for this species.

The effects of four light intensities (1000 lx, 500 lx, 50 lx, 3 lx) on growth, survival and feeding activity in common sole (Solea solea L.) larvae were studied from 4 to 51 days post hatching (dph). During the pelagic larval stage (4–12 dph), larvae reared at 3 lx showed a lower growth. From 19 onwards, the larvae reared under 3 lx displayed a significant (P ≤ 0.05) higher SGR than the other treatments and a higher final weight compared to 1000 lx and 500 lx. Survival rate was higher under intermediate light intensities (500 and 50 lx). Larvae reared at 3 lx displayed a significant delay in the degree of metamorphosis compared to the other treatments, while at 33 dph metamorphosis was completed under all treatments. Histological examination revealed the importance of vision and light in the first feeding of this species, while after metamorphosis, the full development of other sensory organs indicated that feeding activity is also mediated by chemosensory perception. Results indicate that high light intensity seems to be more suitable during the pelagic larvae, while the opposite would ensure better growth from the onset of metamorphosis to the benthic phase.

The present study was carried out to evaluate growth, gut histology and gut bacterial community of gilthead sea bream ( Sparus aurata ) fed with increasing dietary soybean meal (SBM) levels in a low fishmeal (FM) based diet, in comparison with a control diet. Five isoproteic and isolipidic experimental diets were formulated to contain increasing levels of SBM (0, 100, 200, and 300 g kg −1 named S0, S10, S20 and S30, respectively) with 150 g kg −1 of FM, and one control diet (C) without SBM and containing 350 g kg −1 of FM. Sixty sea bream (initial body weight 75.9 ± 1.9 g, n = 900) per tank were reared in a recirculation system at 23.0 ± 1.0 °C and fed to satiation. The trial was run in triplicate and lasted 100 days. At the end of the trial fish fed the S30 diet showed a higher ( P ≤ 0.05) specific growth rate (SGR) compared to S0 (SGR, 1.17 ± 0.03, 1.20 ± 0.01, 1.22 ± 0.01, 1.25 ± 0.01 and 1.21 ± 0.04 for S0, S10, S20, S30 and C, respectively), and a higher feed intake (FI) compared to S0, S10 and S20. Sea bream fed the C diet had a higher ( P ≤ 0.05) FI compared to S0 (FI, 1.40 ± 0.01, 1.45 ± 0.01, 1.44 ± 0.03, 1.51 ± 0.03 and 1.46 ± 0.02 for S0, S10, S20, S30 and C, respectively). No significant differences in feed conversion rate, protein efficiency ratio, gross protein efficiency and gross lipid efficiency among the treatments were detected. No specific histopathological changes indicative of soy-induced enteritis were observed in the intestine of any fish examined. Gut bacterial community of the distal intestine content was analyzed by Next-Generation Sequencing. At the phylum level, the gut bacterial community was dominated by Firmicutes (relative abundance 71%), while the most represented family was Lactobacillaceae (26%). Even if no significant differences ( P ≤ 0.05) in the gut bacterial community α and β-diversity according to the different diets were detected, Cyanobacteria and Lactobacillaceae progressively increased from diet C to diet S30. In conclusion results of growth, nutrient utilization, gut histology and gut bacterial community indicate that SBM can be successfully incorporated up to a level of 300 g kg −1 with the inclusion of 150 g kg −1 of FM, without any deleterious effects on growth, protein utilization and gut health during the on-growing of sea bream.

Changing trends in fish feed formulation, with progressively higher inclusion levels of plant ingredients, are invariably introducing more fibre despite the fact that this component cannot be utilized by most fish. The effects of increasing insoluble dietary fibre level on growth, nutrient utilisation, blood parameters and gut health in European sea bass (Dicentrarchus labrax L.) were studied over a period of 117 days. Moreover, investigation on digesta transit time through gastrointestinal evacuation pattern and digesta characteristics (moisture of digesta) were studied. Five isoproteic diets were formulated to contain increasing insoluble fibre levels, neutral detergent fibre, NDF (7.2, 8.9, 11.5, 13.1 and 15.5%) derived by the inclusion of sunflower hulls and soybean hulls. No significant differences due to fibre inclusion levels were observed in final body weight, specific growth rate, feed intake, feed conversion rate, protein and lipid efficiency. No significant differences in serum total protein, glucose, triglycerides, alkaline phosphatase and inorganic phosphorous were found. All the histological sections showed normal intestinal architecture, and inflammatory and/or degenerative changes were not present in any histological section from all subjects examined. The investigation into gastrointestinal evacuation pattern revealed no significant differences between treatments, however higher dietary fibre levels seem to increase the time required to empty the stomach while the time required to empty 90% of the hindgut content was similar in all the treatments: around 46–47 h. No differences were found between diets in the moisture content of digesta along the digestive tracts. We can conclude that the different insoluble fibre levels tested in this trial have no effects on overall performances and feed efficiency in European sea bass. Results from blood biochemistry profile and histology confirm good nutritional and health status of fish under all feeding treatments. The inclusion of fibre had no influence on digesta transit time. In formulation of feed for the on-growing of European sea bass insoluble fibre derived from sunflower hulls and soybean hulls can be included at a level of up to 15.5%.

The aquaculture industry depends upon the development of sustainable protein sources to replace fishmeal (FM) in aquafeeds and the products derived from soybeans are some of the most studied plant feedstuffs. A key area of investigation for continuing to improve modern aquafeeds includes the evaluation of varying proportions and combinations of plant ingredients to identify mixtures that are more efficiently utilized by the fish. This study investigated the effects of increasing soybean meal (SBM) by replacing a mix of plant ingredients in low FM (20%) diets on growth, blood biochemistry profile and gut histology on European sea bass. Five isonitrogenous and isolipidic experimental diets were formulated: four diets containing increasing SBM levels (0, 10, 20 and 30%; 0SBM, 10SBM, 20SBM and 30SBM, respectively) with a low content of FM (20%) and one control diet (0% SBM; 35% FM). Diets containing SBM brought to comparable performance and protein utilization, while 0SBM had negative impact on feed conversion rate and protein utilization. Blood parameters suggested an optimal nutritional status under all feeding treatments, even though slightly decreased values were reported at increasing dietary SBM. Histology examination did not show any changes indicative of soy-induced enteritis. We can conclude that for European sea bass: (i) different blends of plant protein did not affect feed intake despite the 20% FM dietary level; (ii) the inclusion of SBM maintains optimal growth and feed utilization in low FM diets; (iii) blood biochemistry profile showed a good nutritional status under all feeding regimes; (iv) no evidence of soy-induced enteritis was reported in any group fed low FM diets. For formulation of practical diets in on-growing of European sea bass, SBM up to 30% can be successfully incorporated into feeds containing low FM inclusion.

This article proposes a systematic evaluation of methods pertaining to the pathology discipline (histology, histochemistry, ultrastructure, in situ techniques, image analysis including morphom-etry and fractal dimension analysis) employed to study liver and intestine status during a series of nutritional trials performed on flatfish species (common sole, Solea solea; Senegal sole, Solea senegalensis and turbot, Scophthalmus maximus) of commercial interest. Histology is the first step able to detect the normal architecture of the tissues or the possible occurring changes related to inflammation, degeneration and cell death. The evaluation of liver should include a scoring system to obtain semi-quantitative numerical data that are statistically analysable; histochemisty is a valid tool for identifying the type of cell content, together with ultrastructure that also permits the detection of even mild signs of cell damage. For the evaluation of the intestine, in situ techniques are useful together with fractal dimension analysis to determine the mucosal tro-phism, while ultrastructure can demonstrate cell injury. These considerations were resumed in a methodological flowchart as a valid tool to assess the fish health status when nutritional trials are planned.

The effects of early weaning on the performances and molecular ontogeny of digestive enzyme precursors in common sole (Solea solea) larvae were studied. Four experimental weaning time protocols (WPs) were performed in triplicate. One protocol was used for the control group where larvae were fed live feed until 27. days post-hatching (dph) (WP-27) and weaned with dry feed. The other groups provided only with dry feed (WP-4), live feed until 13. dph (WP-13) and live feed until 18. dph (WP-18), followed by weaning on to the same commercial microdiet (MD) used in the WP-27 group (AgloNorse K/S Tromsø Fiskeindustry A/S & Co., Norway). The trial lasted until 33. dph. The larvae of the WP-4 group displayed a significantly lower survival rate, lower growth increase and a delay in the onset of metamorphosis when compared with the other groups. The WP-13 and WP-18 groups displayed significantly lower weight increases when compared with the WP-27 group, whereas metamorphosis development and survival were not influenced. Partial transcripts of trypsinogen 1 (tryp1), trypsinogen 3 (tryp3), trypsinogen Y (trypY), chymotrypsinogen (chymt), amylase (amy) and hepatic lipase (HL) were cloned and analysed by quantitative real-time PCR (qPCR). The tryp1 and chymt mRNA levels displayed a rapid increase in all the treatments between 4 and 8. dph and thereafter displayed a homogeneous fluctuating pattern with a higher increasing tendency for chymt at the end of the trial. The amount of tryp3 and trypY mRNA remained stable or slightly increased until 27. dph in all WP groups and thereafter displayed a general up-regulation trend. The amy transcript levels displayed a gradual increase throughout the study in all WP groups. The HL mRNA levels increased until 13. dph in the WP-27, WP-18 and WP-13 groups and thereafter remained generally stable until 27. dph, whereas in the WP-4 group, there was a linear time-related increasing trend until the end of the trial. This study supports previous results on the feasibility of early weaning of common sole larvae using a MD before the end of metamorphosis. The molecular ontogeny of some important digestive enzyme precursors is described for the first time in this species. The transcripts analysed were not greatly influenced by the WPs. © 2013 Elsevier B.V., The study was financed by the Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (IZLER, Italy) and by the Spanish Ministry of Economic Affairs and Competitiveness (MINECO), by “Ridigest” (AGL2011-23722) and “AQUAGENOMICS” (Consolider-Ingenio Program; CDS2007-0002) project grants to MY. We also wish to thank Larvanet-COST ACTION FA0801 (EU RTD framework Programme) (www.larvanet.org).

Currently available data show that shellfish and finfish production in Italy, derived both from fisheries and aquaculture activities, is on the order of 474,000 tons, each activity representing 50 % of the total amount. In this context, the finfish aquaculture industry contributes on average 31 % to the national aquaculture production and on average 59 % of its value, giving a total amount of 72,000 tons and a value of around 351 million € (2010). According to FEAP statistics, Italy is the fourth largest finfish producer in EU27, after the UK, Greece, and Spain, while it is also one of the six largest finfish producers among the non- EU and EU member countries, together with Norway, UK, Greece, Turkey, and Spain. Presently, fish culture activities are mainly focused on rainbow trout (Oncorhynchus mykiss, 55.5 %), followed by European sea bass (Dicentrarchus labrax, 13.6 %), gilthead sea bream (Sparus aurata, 12.2 %), gray mullet (Mugil cephalus, 5.3 %), sturgeon (Acipenser spp., 2 %), and European eel (Anguilla anguilla, 1.7 %). Over the last 20 years, freshwater fish production and aquaculture (trout, carp, and eel) have decreased in Italy, with the exception of sturgeon. In contrast, marine fish production has significantly increased during the same period, and the two leading species, European sea bass and gilthead sea bream, presently contribute 25.8 % of the finfish production. From 1,900 tons in 1990, production reached 19,000 tons in 2010, with a 900 % increase, at an average percentage of 4.5 %. In addition, new marine fish species were successfully cultured over the same period. This review outlines the past and present situation of finfish culture in Italy and discusses future developments and priorities, with particular emphasis on new, emerging aquaculture species.

In vertebrates, chemosensitivity of nutrients occurs through the activation of taste receptors coupled with G-protein subunits, including α-transducin (G(αtran)) and α-gustducin (G(αgust)). This study was aimed at characterising the cells expressing G(αtran) immunoreactivity throughout the mucosa of the sea bass gastrointestinal tract. G(αtran) immunoreactive cells were mainly found in the stomach, and a lower number of immunopositive cells were detected in the intestine. Some G(αtran) immunoreactive cells in the stomach contained G(αgust) immunoreactivity. Gastric G(αtran) immunoreactive cells co-expressed ghrelin, obestatin and 5-hydroxytryptamine immunoreactivity. In contrast, G(αtran) immunopositive cells did not contain somatostatin, gastrin/cholecystokinin, glucagon-like peptide-1, substance P or calcitonin gene-related peptide immunoreactivity in any investigated segments of the sea bass gastrointestinal tract. Specificity of G(αtran) and G(αgust) antisera was determined by Western blot analysis, which identified two bands at the theoretical molecular weight of ~45 and ~40 kDa, respectively, in sea bass gut tissue as well as in positive tissue, and by immunoblocking with the respective peptide, which prevented immunostaining. The results of the present study provide a molecular and morphological basis for a role of taste-related molecules in chemosensing in the sea bass gastrointestinal tract.

The in vitro effect of a Yucca schidigera extract (YSE) and tannins from chestnut wood on composition and metabolic activity of canine and feline faecal microbiota was evaluated. Four treatments were carried out: control diet, chestnut tannins (CT), YSE and CT + YSE. The YSE was added to canine and feline faecal cultures at 0.1 g/l, while CT were added at 0.3 g/l for a 24-h incubation. A total of 130 volatile compounds were detected by means of headspace-solid phase microextraction gas-chromatography/mass spectrometry analyses. Several changes in the metabolite profiles of fermentation fluids were found, including a decrease of alcohols (-19%) and esters (-42%) in feline and canine inoculum, respectively, which was due to the antibacterial properties of tannins. In canine inoculum, after 6 h, YSE + CT caused lower cadaverine concentrations (-37%), while ammonia (-4%) and quinolone (-27%) were reduced by addition of CT. After 24 h, the presence of CT resulted in a decrease of sulphur compounds, such as dimethyl sulphide (-69%) and dimethyl disulphide (-20%). In feline faecal cultures, after 6 h, CT lowered the amount of indole (-48%), whereas YSE tended to decrease trimethylamine levels (-16%). Both in canine and feline inoculum, addition of CT and, to a minor extent, YSE affected volatile fatty acids patterns. In canine faecal cultures, CT exerted a marginal inhibitory effect on Escherichia coli population (-0.45 log 10 numbers of DNA copies/ml), while enterococci were increased (+2.06 log 10 numbers of DNA copies/ml) by YSE. The results from the present study show that YSE and tannins from chestnut wood exert different effects on the composition and metabolism of canine and feline faecal microbiota. In particular, the supplementation of YSE and tannins to diets for dogs and cats may be beneficial due to the reduction of the presence of some potentially toxic volatile metabolites in the animals' intestine.

A 9-week feeding trial was performed to evaluate the effects of fishmeal (FM) replacement by a mixture of plant proteins (PP), consisted of wheat gluten, soybean meal and soy protein concentrate, on the growth performance and welfare of turbot juveniles (initial weight 9.7 ± 0.2 g). Four isonitrogenous and isolipidic diets contained FM at 500 g kg−1 (FM50), 350 g kg−1 (FM35), 200 g kg−1 (FM20) and 50 g kg−1 (FM5). A decreased feed intake and an increased feed conversion rate was observed in FM5 group. Specific growth rate was significantly reduced in FM20 and FM5 groups, whereas protein and lipid utilization and proximate whole body composition were significantly different in FM5 group. Serum cortisol significantly increased in FM20 and FM5 groups whereas cholesterol, triglycerides, NEFA, total protein and urea concentrations significantly decreased. Serum lysozyme and blood phagocytes increased in FM20 and FM5 groups. FM35 ensured growth close to FM50, without significant effects on health and welfare of animals. FM20 and FM5 groups displayed reduced growth, metabolic stress and an immune response with effects on health and welfare. Results highlighted the consistency between growth performance and welfare status, suggesting the usefulness of their combined assessment for evaluating the suitability of PP and to improve dietary formulation for turbot.

Vertebrates perceive a variety of exogenous substances using two main chemosensory systems, taste and olfaction. The taste perception occurs through the interaction of taste receptors associated with specific G protein subunits such as a-transducin (Gatran) and a-gustducin (Gagust). Aquatic vertebrates are also provided with a chemosensory system consisting of solitary chemosensory cells distributed to the oropharynx and skin. In this study, we identified Gatran and Gagust-immunoreactive cells intermingled with non-labeled epithelial cells in the gastric mucosa of the common sole. A long-term diet with increasing concentrations of mussel meal in the protein component of a conventional fish meal-based diet induced a dose-dependent increase in the gastric epithelial area and density of Gatran and Gagust immunoreactive cells. These findings suggest that taste-related molecules are regulated by changes in diet formulation in common sole aquaculture.

Fish are free-living organisms since initial stages of development and are exposed to numerous pathogens before their lymphoid organs have matured and adaptive immunity has developed. Susceptibility to diseases and juvenile mortality represent key critical factors for aquaculture. In this context, the characterization of the appearance kinetics of the immune system key members will be useful in understanding the ability of a particular species in generating immune protection against invading pathogens at different developmental stages. The present study characterized, for the first time, the transcriptional onset of un-explored relevant genes of both innate and adaptive immune system during the Solea solea ontogenesis. Gene expression profiles of immune relevant genes was investigated, by means of DNA microarray, in ten developmental stages, from hatching (1 day post-hatching, dph) to accomplishment of the juvenile form (33 dph). The obtained results revealed that transcripts encoding relevant members of innate immune repertoire, such as lysozyme, AMPs (hepcidin, β-defensin), PPRs and complement components are generally characterized by high expression levels at first stages (i.e. hatch and first feeding) indicating protection from environmental pathogens even at early development. Transcription of adaptive immune genes (i.e. Class I and class II MHC, TCRs) differs from that of the innate immune system. Their onset coincides with metamorphosis and larvae-to-juvenile transition, and likely overlaps with the appearance and maturation of the main lymphoid organs. Finally, data collected suggest that at the end of metamorphosis S. solea cell-mediated immune system hasn't still undergone full maturation.

The possibility of increasing n-3 and n-6 long-chain polyunsaturated fatty acids (PUFA) content in microalgal mixtures used to feed Tapes philippinarum larvae was explored by lowering culture temperature from 26 to 14 °C. Although fatty acid composition of different microalgal species has a genetic basis, the algal cultures grown at 14 °C significantly increased the content of long-chain n-3 PUFA in Isocrysis galbana and in Thalassiosira pseudonana, while in Tetraselmis tetrathelo, the PUFA increase only involved shorter chain PUFA, namely 16:4n-3 and 18:4n-3. However, larvae fed on the PUFA enriched microalgal mixture did not show improvements in growth and survival performances with respect to the control group fed the microalgal mixture grown at 26 °C. From a biochemical perspective, two key aspects emerged from the results: (i) clam larvae have adequate biotransformation and selection skills to adjust fatty acid profile to their requirements as they can even modulate the incorporation of essential long-chain PUFA as 20:5n-3 and 22:6n-3 when the dietary supply exceeds the physiological requirements; (ii) bivalve can biosynthesize non-methylene-interrupted dienoic (NMID) fatty acids as confirmed by the constancy of relative proportion with larvae growth in spite of the NMID fatty acid absence in the diet.

Three isoproteic (47% protein) diets were formulated to contain graded levels of crude fat (diet D16:16%, diet D24: 24% and diet D32: 32%). Each diet was fed to satiation in three and to 80% satiation in two replicate groups of gilthead sea bream (Sparus aurata), having an initial body weight of 72–74 g. The trial lasted 81 days. Groups fed to satiation showed higher final body weight (FBW; 238.8–252.3 g vs. 218.0–229.3 g) and daily growth index (DGI; 2.49–2.65%/day vs. 2.27–2.34%/day) than those fed to 80% satiation. Feed intake was significantly different both for feeding level and for diet composition. Fish fed to satiation had higher feed conversion rate (FCR) compared to the 80% satiation groups (1.33–1.44 vs. 1.13–1.17; P B 0.001). Within satiation groups, FCR was significantly lower in fish fed D16 compared to fish fed D32 (1.33 vs. 1.44, P B 0.05), whereas no statistical differences were found within the 80% satiation groups. The increase in dietary lipid level did not improve growth performance, feed efficiency and protein utilization but decreased gross lipid efficiency. Conversely, a reduction in ration from satiation to 80% satiation decreased DGI, thus improving FCR. Feed costs were influenced by dietary energy level and feeding ratio, the lowest energy diet at 80% satiation being the most profitable combination among the variables.

The effects of feeding 1,3/1,6 β-glucans on the innate and the adaptive immune responses of European sea bass (Dicentrarchus labrax) was investigated. Two experiments were carried out during the study. In the first, a number of non-specific immune parameters were examined at 4, 7, 10, 14, 21 and 25 days of feeding fish with a semipurified diet containing Macrogard ©, a commercially available form of 1,3/1,6 β-glucans. The respiratory burst activity of head kidney macrophages isolated from the different groups of fish fed the immunostimulant peaked and subsequently decreased at different times during the experiment. Head kidney macrophages from fish fed 250 ppm β-glucans had a statistically higher level of respiratory burst activity at Day 21 of feeding compared with fish fed no immunostimulant. No statistical differences were observed in lyzozyme activity during this trial. In the second experiment, the effect of feeding 1,3/1,6 β-glucans on the immune response of fish to an alginate-encapsulated Vibrio vaccine administered orally was examined. Respiratory burst of head kidney macrophages and serum lysozyme activity decreased in all fish over the course of the trial, while serum lysozyme activity was considerably lower than values obtained in the first experiment. Fish vaccinated orally had significant increases in antibody response by Week 2 post-vaccination, but β-glucans did not appear to affect these levels. Vaccination may have resulted in activating the immune system as a whole, thus masking any difference in immunostimulation by the β-glucans. It may be that the optimal doses and timing of β-glucans administration is different when the immunostimulant is administered alone or in combination with the vaccine. In conclusion, European sea bass can be immunomodulated with oral administration of β-glucan. Optimal doses and administration times have been established when β-glucans are fed alone, although further studies are needed to establish ideal feeding regimes for glucans administrated in combination with vaccination. Questo studio è stato condotto per determinare l’effetto che i 1,3/1,6 β-glucani possono esercitare sul sistema immunitario specifico e aspecifico del branzino Dicentrarchus labrax. A questo scopo sono stati effettuati due esperimenti. Nel primo, alcuni parametri relativi al sistema immunitario aspecifico sono stati esaminati dopo 4, 7 10, 14, 21 e 25 giorni di alimentazione con diete semi-purificate contenenti il prodotto commerciale a base di β-glucani Macrogard®. L’attività relativa allo scoppio respiratorio dei macrofagi del rene craniale isolati dagli animali di tutti i gruppi sperimentali ha mostrato un picco di attività seguito da un decremento, ma con temporalità differenti a seconda del dosaggio. I macrofagi dei pesci alimentati con la dieta contenente 250 ppm di glucani hanno mostrato differenze statisticamente significative rispetto agli altri gruppi al giorno 21 di somministrazione. Nessuna differenza è stata riscontrata per quanto riguarda l’attività del lisozima. Nel secondo esperimento, gli effetti della somministrazione orale di β-glucani sono stati valutati quando somministrati insieme ad un vaccino orale contro la vibriosi. L’attività relativa allo scoppio respiratorio dei macrofagi e l’attività del lisozima sono calati lungo tutto il periodo di osservazione, con quest’ultimo valore considerevolmente più basso rispetto a quello ottenuto nel primo esperimento. I pesci vaccinati oralmente hanno mostrato un aumento della risposta anticorpale dopo 2 settimane dalla somministrazione di vaccino anche se i β-glucani non hanno mostrato effetti sul sistema immunitario specifico dei pesci. La vaccinazione, stimolando l’attività del sistema immunitario, può aver mascherato l’effetto dei β-glucani. La dose ottimale di β-glucani potrebbe essere differente quando questi vengono somministrati insieme ad un vaccino. In conclusione, il branzino sembra poter essere immunomodulato mediante la somministrazione oral dei β-glucani. È stato possibile stabilire la dose e il tempo di somministrazione ottimali quando i β-glucani vengono utilizzati singolarmen-te; al contrario, sono necessari ulteriori studi per stabilire la dose ottimale quando questi vengono somministrati insieme ad un vaccino.

Thepresentstudywasperformedtodeterminetheef-fect of soybean meal (SBM) on the performance andguthistologyofgiltheadseabreamandEuropeanseabass. Three isonitrogenous and isolipidic extrudeddiets (crude protein, 470gkg 1 diet; crude fat,200g kg 1 diet) were formulated containing 0(0SBM), 180 (180SBM) and 300 (300SBM)gkg 1 diet SBM and tested on both species in two separateexperiments. Fish at an initial average weight ofaround18gwererandomlyallocatedto800Lsquaretanksconnectedtoaclosedrecirculatingsystem.Thetrialslasted80daysforseabreamand89daysforseabass. Fish were hand-fed to apparent satiation. In-creasing the level of SBM had no signi¢cant eiectsonthe speci¢cgrowth rate, feed intakeand feedcon-versionrateinboththespecies. Intheseabreamdis-tal intestine, lamina propria was moderately anddiiuselyexpanded in some ¢shduetoan increase incell in¢ltration represented by mononuclear cells,this ¢nding being more frequent in animals fed diet300SBM. No other morphological alterations in in-testinal folds, enterocytes or other in£ammationsigns were noticed in the sea bream distal intestineof any group. No histological diierences were foundinseabassinanyexperimentalgroup.Keywords: gilthead sea bream, European seabass, soybean meal, growth, gut histologyIntroductionCurrently, the importance of vegetable protein inaquaculture is well recognized by feed industry op-erators due to the growing pressure to ¢nd ¢sh meal(FM) substitutes in ¢sh diets. Among the diierentingredients, soybean meal (SBM) is one of the mostinterestingalternativeFMbecauseof theadvantagesof supply, price and protein and amino acid (AA)composition. On the other hand, the use of soybeanproducts may pose a series of problems that mayaiect palatability (Rackis 1974), protein digestibility(Glencross, Carter, Duijster, Evans, Dods, McCaierty,Hawkins, Maas & Sipsas 2004) and phosphorus bio-availability (Vielma, Makinen, Ekholm & Koskela2000), and may reduce feedutilization. Diierent ¢shspecieshaveshownvariablesensitivitytosoyantinu-tritional factors. In Atlantic salmon and rainbowtrout,SBMcausesimpairedgrowthandproteinutili-zation in addition to distinct morphological altera-tions in the intestine that become more pronouncedwith increasing dietary level (Krogdahl, Bakke-McKellep & Baeverfjord 2003). The morphologicalalterations are characterized by a widening of thelaminapropria,whichisin¢ltratedbyamixedpopula-tion of in£ammatory cells, and concurrent changesin enterocyte structure, including a decrease in oreven a loss of the supranuclear vacuoles present innormalenterocytes (Baeverfjord & Krogdahl1996), ashorteningof the microvilliand increased formation

International audience; Coastal systems could be affected by hypoxic events brought about by global change. These areas are essential nursery habitats for several fish species including the common sole (Solea solea L.). Tolerance of fish to hypoxia depends on species and also on their physiological condition and nutritional status. Indeed, high dietary lipid content has been recently shown to negatively impact the resistance of sole to a severe hypoxic challenge. In order to study the molecular mechanisms involved in the early response to hypoxic stress, the present work examined the hepatic transcriptome in common sole fed diets with low and high lipid content, exposed to severe hypoxia. The activity of AMP-activated protein kinase (AMPK) was also investigated through the quantification of threonine-172 phosphorylation in the alpha subunit. The results show that hypoxia consistently regulates several actors involved in energy metabolism pathways and particularly AMPKα, as well as some involved in cell growth and maintenance or unfolded protein response. Our findings reveal that (1) the expression of genes involved in biological processes with high energy cost or implicated in aerobic ATP synthesis was down-regulated by hypoxia, contrary to genes involved in neoglucogenesis or in angiogenesis, (2) the consumption of high lipid induced regulation of metabolic pathways going against this energy saving, and (3) this control was fine-tuned by the regulation of several transcriptomic factors. These results provide insight into the biological processes involved in the hepatic response to hypoxic stress and underline the negative impact of high lipid consumption on the tolerance of common sole to hypoxia.

Summary The present study was carried out to test different mussel meal (MM) dietary levels in combination with fishmeal (FM) on the growth performance, fatty acid composition and liver histology of common sole, Solea solea juveniles to highlight the growth potential of this species. Four isoproteic (53%) and isolipidic (11%) pelletized diets were formulated to contain graded levels of mussel meal, MM0 (0%), MM25 (25%), MM50 (50%) and MM75 (75%), up to 75%. Sole juveniles (initial individual mean body weight 13.1 ± 2.3 g, n = 840) were fed to satiation for 91 days. Seventy fish per tank (500-L, 0.64 m2 bottom surface) were reared in 12 tanks (3 tanks per treatment) at 20 ± 1°C. Diets containing MM (MM25, MM50 and MM75) gave a significantly higher specific growth rate (SGR, 1.27 ± 0.01, 1.38 ± 0.06 and 1.40 ± 0.05, respectively), higher feed intake and lower feed conversion rate (FCR, 1.09 ± 0.01, 1.00 ± 0.04 and 0.98 ± 0.02, respectively) when compared to the FM-based diet (MM0, SGR, 0.98 ± 0.11, FCR, 1.52 ± 0.13). Carcass proximate composition was not influenced by dietary treatments, with the exception of the significantly lower lipid content in the MM75 group. Protein efficiency ratio (PER) and gross protein efficiency (GPE) were significantly improved by the mussel meal inclusion (PER, 1.29 ± 0.12, 1.76 ± 0.01, 1.89 ± 0.06, 1.95 ± 0.08; GPE, 25.29 ± 1.85, 33.38 ± 0.89, 35.96 ± 1.36, 36.59 ± 1.05 in MM0, MM25, MM50 and MM75, respectively). A significant decrease in the viscerosomatic index was observed in fish fed with MM50 and MM75 in comparison to MM0. The hepatosomatic index of fish fed with MM0 and MM25 was higher than that of fish fed with MM75, although the histological examination of liver parenchyma in all experimental groups showed a uniformly abundant accumulation of lipid droplets. Carcass fatty acid composition was significantly affected by dietary treatments, reflecting the dietary fatty acid profile. According to these results, the inclusion of MM in experimental FM-based diets improved the performance and feed utilization of common sole juveniles. The inclusion of MM in the present trial allowed a higher SGR than that registered in previous growth trials on common sole. This study could provide useful information to detect effective ingredients for practical diets in Solea solea. It also seems advisable to consider an inclusion of at least 25% MM in the experimental reference diet to be used for further application towards the development of specific diets for this species.

Knowledge about the nutritional requirements of common sole ( Solea solea ) is limited and no information regarding the optimal dietary lipid level is available yet. Thus, this study was undertaken to assess the growth response and feed utilization of common sole juveniles fed diets with increasing lipid levels. Four isonitrogenous (59% protein) pelletized diets with different dietary lipid levels (8%, 12%, 16% and 20%; L8, L12, L16 and L20, respectively) were fed to triplicate fish groups of 80 individuals to apparent satiation over 150 days. A one-way ANOVA, Tukey's post hoc test and linear regression were used to analyse the data ( P ≤ 0.05). At the end of the trial, final body weight was significantly higher in fish fed L8 (40.7 ± 1.7 g), followed by those fed L12 (35.1 ± 1.2 g), L16 (27.9 ± 2.5 g) and L20 (22.1 ± 0.3 g). The specific growth rate was higher in fish fed L8 and L12, compared to the other treatments, and it was lowest in L20. Voluntary feed intake decreased with an increase in the dietary lipid level. The feed conversion rate, the protein efficiency ratio and the gross protein efficiency were lower in fish fed L20, while no significant differences were observed among L8, L12 and L16. Gross lipid efficiency was significantly higher in fish fed low lipid diets. Histological observations showed that 19 of 36 observed subjects had lipid droplets in the cytoplasm of enterocytes at the apex of the mucosal folds and, in some cases, also along the entire fold (intestinal steatosis). The number of fish with intestinal steatosis in groups L20 and L16 was significantly higher than the number in group L8. Ultrastructure showed large electrondense lipid droplets within the cytoplasm of enterocytes and warping of the cytoplasmic membrane (steatosis); in some cases, lipid droplets were also present within the Golgi apparatus. In conclusion, the results of this trial suggest that the diet for S. solea juveniles should include no more than 12% lipids. Higher lipid inclusions not only led to a substantial decline in performance but also affected gut health. This should be taken into consideration in formulating specific practical diets for common sole.

The present study was undertaken in order to determine the effect of a dietary incorporation of soybean meal (SBM) on growth performance, nutrient utilization and gut histology of Egyptian sole Solea aegyptiaca. This species, which is genetically and morphologically close to Solea solea, is currently present in the Mediterranean Sea and reared in some Italian farms, using the same techniques for S. solea production and it could represent a promising new species for Mediterranean aquaculture. Three isoproteic and isolipidic extruded diets (crude protein, 47%, crude fat, 20%) were formulated containing 0 (diet 1), 18 (diet 2) and 30% SBM (diet 3). 540 juveniles (initial body weight of 6.9 ± 0.2 g) were equally distributed into nine 150 l squared tanks (bottom surface: 520 cm2) connected with a recirculation system (temperature 20 ± 1 °C and dissolved oxygen above 6 ppm). Animals have been hand-fed twice a day (at 9.00 a.m. and 5.00 p.m.) at a daily ratio of 1.5% body weight− 1. Diets were tested in triplicate for 87 days. The fish were weighed at days 28, 57 and 87, and they were sampled at days 0, 57 and 87 for nutrient retention determination. One-way ANOVA with Newman–Keuls' post-test were used to analyze data (P ≤ 0.05). No differences in terms of palatability were observed and any diet has been consumed completely all throughout the experiment. At day 87, fish fed diet 1, 2 and 3 reached weights of 25.8 ± 0.8 g, 27.5 ± 0.9 g and 26.1 ± 1.5 g, respectively and no differences in performances or nutrient retention were found. Histopathological gut examinations have revealed no noticeable differences in the appearances of the intestines between any of diet groups. Intestinal mucosal cells presented well vacuolated upper, with well defined microvillar brush borders. On the basis of the results of this trial, SBM seems to be a good protein source for Egyptian sole and can be added in the diet up to 30% without any reduction in growth rate and no adverse effect on gut histology.