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Modern aquaculture feeds tend to contain lower levels of fish based ingredients, while increasing the content of plant ingredients. However, this may alter the vitamin profile of the feeds, leading to unbalanced vitamin supply. Requirements for several vitamins have been established for species such as carps and salmonids, but adequate levels for gilthead sea bream are yet unknown. Vitamin D is mainly involved in Ca homeostasis by regulating Ca uptake and liberation from bone intervening in bone remodeling. Fish are unable to synthesize vitamin D and so require absorbing it directly from the diet, thus, it is considered essential for fish. A practical plant-based diet containing 10% fish meal and 6% fish oil containing five levels of vitamin D3 (0.15, 0.43, 0.50, 0.55 and 0.65 mg kg− 1 or 5.8, 17.0, 20.0, 22.0 and 26.0 IU g− 1 ) were formulated to identify the optimum levels for gilthead seabream juveniles. Feeding juveniles of gilthead seabream with a range of vitamin D3 levels between 5.8 and 26.0 IU g− 1 for 70 days did not markedly alter growth. Increase dietary vitamin D3 significantly raised the liver contents in vitamin D3 in a dose-dependent manner following a potential regression. Increased dietary vitamin D3 levels up to 11.6 IU g− 1 may reduce the incidence of skeletal anomalies, particularly caudal and maxillary anomalies, whereas further elevation of dietary vitamin D3 levels increased the concentration of vitamin D3 in liver as well as skeletal anomalies in association to the up-regulation of alp and bmp2 gene expression. The occurrence of myocarditis signs in fish fed vitamin D3 levels of 20.0 IU g− 1 or more denote the toxic effects of these dietary levels. These results, together with the increased occurrence of skeletal anomalies in seabream fed the highest dietary vitamin D3 levels, suggest initial signs of hypervitaminosis D. Thus, the recommended level for vitamin D3 for gilthead seabream juveniles fed diets containing high levels of plant ingredients was suggested to be 11.6 IU g− 1.

An effective replacement for fish meal (FM) and fish oil (FO) based on plant-based raw materials in the feed of marine fish species is necessary for the sustainability of the aquaculture sector. However, the use of plant-based raw materials to replace FM and FO has been associated with several negative health effects, some of which are related to oxidative stress processes that can induce functional and morphological alterations in mucosal tissues. This study aimed to evaluate the effects of dietary oligosaccharides of plant origin (5,000 ppm; galactomannan oligosaccharides, GMOS) and a phytogenic feed additive (200 ppm; garlic oil and labiatae plant extract mixture, PHYTO) on the oxidative stress status and mucosal health of the gills of juvenile European sea bass (Dicentrarchus labrax). The experimental diets, low FM and FO diets (10%FM/6%FO) were supplemented with GMOS from plant origin and PHYTO for 63 days. GMOS and PHYTO did not significantly affect feed utilization, fish growth, and survival. GMOS and PHYTO downregulated the expression of β-act, sod, gpx, cat, and gr in the gills of the fish compared with that in fish fed the control diet. The expression of hsp70 and ocln was upregulated and downregulated, respectively, in the GMOS group compared with that in the control group, whereas the expression of zo-1 was downregulated in the PHYTO group compared with that in the GMOS group. The morphological, histopathological, immunohistochemical, and biochemical parameters of the fish gills were mostly unaffected by GMOS and PHYTO. However, the PHYTO group had lower incidence of lamellar fusion than did the control group after 63 days. Although the tissular distribution of goblet cells was unaffected by GMOS and PHYTO, goblet cell size showed a decreasing trend (−11%) in the GMOS group. GMOS and PHYTO significantly reduced the concentration of PCNA+ in the epithelium of the gills. The above findings indicated that GMOS and PHYTO in low FM/FO-based diets protected the gill epithelia of D. labrax from oxidative stress by modulating the expression of oxidative enzyme-related genes and reducing the density of PCNA+ cells in the gills of the fish.

Simple Summary The present study was on the gene expression of a rate-limiting enzyme in long chain polyunsaturated fatty acids (LC-PUFAs), fatty acyl desaturase 2 (fads2), throughout the embryonic development of a gilthead sea bream. The results showed a maternal transfer of fads2 mRNA to the developing oocyte. The embryonic fads2 expression might start after the neurula stage. No effect was found in fads2 expression in developing eggs from broodstock fed with a diet rich in rapeseed oil or fish oil. The present study provides information on the change of LC-PUFA biosynthesis during embryogenesis. Abstract Previous studies have shown that it is possible to increase the ability of marine fish to produce long-chain polyunsaturated fatty acid from their 18C precursors by nutritional programming or using broodstock with a higher fatty acyl desaturase 2 (fads2) expression. However, those studies failed to show the effect of these interventions on the expression of the fads2 gene in the developing egg. Moreover, there were no studies on the temporal expression of the fads2 during ontogeny in the gilthead sea bream (Sparus aurata). In order to determine the changes in expression of fads2 during ontogeny, gilthead sea bream broodstock with a high (HRO) or low (LRO) fads2 expression fed a diet previously used for nutritional programming, or a fish oil-based diet (LFO) were allowed to spawn. The samples were taken at the stages of spawning, morula, high blastula, gastrula, neurula, heart beating, hatch and 3 day-old first exogenous feeding larvae to determine fads2 expression throughout embryonic development. The results showed the presence of fads2 mRNA in the just spawned egg, denoting the maternal mRNA transfer to the developing oocyte. Later, fads2 expression increased after the neurula, from heart beating until 3-day-old larvae, denoting the transition from maternal to embryonic gene expression. In addition, the eggs obtained from broodstock with high fads2 expression showed a high docosahexaenoic acid content, which correlated with the downregulation of the fads2 expression found in the developing embryo and larvae. Finally, feeding with the nutritional programming diet with the partial replacement of fish oil by rapeseed oil did not affect the long chain polyunsaturated fatty acid (LC-PUFA) contents nor fads2 expression in the gilthead sea bream developing eggs.

Elevated mortality during early stages and high occurrence of skeletal anomalies are major concerns in Greater amberjack (Seriola dumerili) juvenile's production. Identification of nutritional requirements during live prey feeding stages, particularly those related with n-3 HUFA would help to formulate specific diets for this species improving larval survival and reducing skeleton anomalies. In this study S. dumerili larvae were reared from 17 to 35 days post hatching (dph). Larvae were fed Artemia sp. containing five different n-3 HUFA levels from 2.7 to 20.5% TFA. Growth, survival, skeletal anomalies occurrence and larval biochemical composition were determined. Best growth, final survival and survival after air stress test were achieved in the range of dietary n-3 HUFA concentrations, between 5 and 20%TFA. The lowest occurrence of skeletal anomalies was achieved in fish fed 12% n-3 HUFA content in Artemia, and the occurrence of cranial anomalies was correlated to increased dietary n-3 HUFA levels. Besides, whole body larval fatty acid profiles showed a positive correlation with dietary n-3 HUFA. Based on the overall results the recommended n-3 HUFA dietary level during Artemia feeding for larval S. dumerili was suggested between 12 and 17%TFA.

The establishment of well-balanced diets that meet nutrient requirements is important to optimize a large-scale production of new aquaculture species. This is the case of meagre (Argyrosomus regius), a promising new aquaculture species, with great potential owing to its high growth rate, feed efficiency and easy adaptation to captivity. Knowledge on the nutritional requirements of this species is still scarce, namely regarding essential fatty acids, which are required to sustain growth, development, immune status and survival. A feeding trial was performed with meagre fingerlings (2.8 g ± 0.23) testing 5 increasing dietary n-3 LC-PUFA levels (0.8, 1.4, 2.0, 2.6 and 3.6% DM) with the purpose of evaluating the n-3 LC-PUFA requirements for fast growth of meagre fingerlings. Meagre reflected very high specific growth rates (4.1 to 4.6%) and low feed conversion ratios (0.7 to 0.8), thus highlighting its great potential for aquaculture production. Fish fed 0.8% n-3 LC-PUFA showed the lowest growth, which was significantly improved by increasing the dietary n-3 LC-PUFA levels up to 2.0–2.6%. DHA and ARA were preferentially retained over EPA in whole fish body. Fish fed 0.8% n-3 LC-PUFA showed an up-regulation of fads2 and elovl5 relative gene expressions. Thus, meagre seems to have active Δ6 desaturases and Elovl5, but their activities being insufficient to produce DHA and EPA from PUFA precursors to sustain fast growth, at least under the experimental conditions tested. Young meagre shows a typical marine requirement for n-3 LC-PUFA, estimated to be, at least, 2.0% DM of the diet.

A sustainable growth of the aquaculture sector implies the use of sustainable novel raw materials as replacers of the traditional fishmeal (FM) and fish oil (FO) ingredients. This fact has led to the development of sustainable and functional diets as part of a management strategy to reduce the effects on fish growth performance and health derived from low FM/FO dietary contents. In this sense, Antarctic krill (Euphausia superba) is considered a potential candidate in dietary inclusions to potentiate fish growth and health status. In this study, European sea bass (Dicentrarchus labrax) were fed a practical diet with either a 15% fishmeal content (KM0; control diet) or the same diet substituted by 30% (KM5; 50 g KM/kg diet) or 50% (KM7.5; 75 g KM/kg diet) Antarctic krill meal (KM) for 12 weeks in triplicates. At the end of the feeding trial, growth performance, liver morphology, liver proximate composition, lipid classes and fatty acid profiles, as well as the expression of hepatic genes related with lipid metabolism were evaluated. Fish fed KM-based diets presented higher (p 0.05) biochemical composition and fatty acid profile. However, smaller hepatocellular area and lower grade of cytoplasm vacuolization as well as a better alignment around sinusoidal spaces were found. The analyses of liver lipid classes revealed a positive correlation between the level of dietary KM and the pigmented material such as astaxanthin and free fatty acid content, as well as a negative correlation with the cholesterol levels. The expression of hepatic genes studied demonstrated a downregulation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (hmgr) and delta-6-desaturase (fads2) expression levels in fish fed KM-based diets. Besides, gene expression levels of fatty acid binding protein 7 (fabp7) and lipoprotein lipase (lpl) were significantly correlated with KM dietary levels. Altogether, these results profile KM as a potential promoter of growth and liver health in European sea bass fed low fish meal and oil diets.

Five isoproteic (54.8%) and isolipidic (24.1%) microdiets, which varied in their docosahexaenoic acid (DHA) content (0.25%, 0.75%, 1.64%, 1.99% and 3.17%; dw), were manufactured to determine its effects on longfin yellowtail Seriola rivoliana larvae in terms of fish biological performance, whole body fatty acid profile and incidence of skeletal anomalies from 30 dah (11.31 ± 1.79 Total Length, TL) to 50 dah (19.80 ± 0.58 mm TL). The inclusion of dietary DHA up to 3.17% (dw) improved larval resistance to air exposure, although DHA did not significantly affect fish final growth or final survival. Indeed, high levels of dietary DHA (1.99% and 3.17%, dw) tended to increase the incidence of skeletal anomalies in S. rivoliana larvae, albeit no significant differences were observed. Furthermore, the occurrence of severe anomalies such as kyphosis and lordosis, was mainly associated to the larvae fed the highest levels of dietary DHA. In terms of survival, increasing dietary DHA levels did not significantly affect longfin yellowtail survival rate, despite a tendency for enhanced survival. The results of the present study proved that the inclusion of dietary DHA in inert diets up to a 3.17% (dw) and a DHA/EPA ratio above 3.1 increased the final survival and stress resistance in S. rivoliana larvae.

11 pages, 6 figures, 1 table, In order to successfully diversify Mediterranean aquaculture, it is necessary to determine optimum culture conditions of potential candidate species such as greater amberjack (Seriola dumerili). Among culture conditions, rearing temperature is a key factor for achieving optimum growth and maintaining fish welfare. However, little is known about the optimum culture conditions of greater amberjack (Seriola dumerili). Thus, the aim of this study was to determine the effect of three different rearing temperatures (17, 22 and 26°C) during 120 days on growth performance, body morphometry, biochemical composition, gut transit and liver morphology of greater amberjack (Seriola dumerili) juveniles. After 120 days of rearing, fish raised at 26°C showed higher (p < .05) body weight and specific growth rate than fish held at lower temperatures, as well as improved feed utilization, protein efficiency and nutrient retention percentages. Fish stomach emptying was faster (p < .05) in fish raised at 26°C than in fish held at 22°C and 17°C. Similar results were obtained for gut transit time, being gut emptying faster (p < .05) in fish reared at 26°C than in fish cultured at lower temperatures. Rearing temperature also induced changes in fish morphology which resulted in a higher (p < .05) caudal propulsion efficiency index for fish reared at 26°C. Based on these results, we conclude that greater amberjack fingerlings perform better at 26°C than at 22°C or 17°C, This work has been funded by the EU seventh Framework Programme by the DIVERSIFY project 7FP‐KBBE‐2013‐GA‐602131

The use of terrestrial raw materials to replace fish meal (FM) and fish oil (FO) in marine fish diets may affect fish growth performance and health. In the last years functional additives have been profiled as good candidates to reduce the effects on health and disease resistance derived from this replacement, via reinforcement of the fish immune system. In the present study, three isoenergetic and isonitrogenous diets with low FM and FO (10% and 6% respectively) were tested based on supplementation either with 0.5% galactomannanoligosaccharides (GMOS diet) or 0.02% of a mixture of essential oils (PHYTO diet), a non-supplemented diet was defined as a control diet. Fish were fed the experimental diets in triplicate for 9 weeks and then they were subjected to a stress by confinement as a single challenge (C treatment) or combined with an experimental intestinal infection with Vibrio anguillarum (CI treatment). Along the challenge test, selected stress and immunological parameters were evaluated at 2, 24 and 168h after C or CI challenges. As stress indicators, circulating plasma cortisol and glucose concentrations were analyzed as well as the relative gene expression of cyp11b hydroxylase, hypoxia inducible factor, steroidogenic acute regulatory protein, heat shock protein 70 and heat shock protein 90 (cyp11b, hif-1α, StAR, hsp70 and hsp90). As immune markers, serum and skin mucus lysozyme, bactericidal and peroxidase activities were measured, as well as gene expression of Caspase-3 (casp-3) and interleukin 1β (il-1ß). The use of functional additives induced a significant (p 0.05) reduction of circulating plasma cortisol concentration when confinement was the unique challenge test applied. Supplementation of PHYTO induced a down-regulation of cyp11b, hif-1α, casp-3 and il-1β gene expression 2h after stress test, whereas StAR expression was significantly (p 0.05) up-regulated. However, when combination of confinement stress and infection was applied (CI treatment), the use of PHYTO significantly (p 0.05) down-regulated StAR and casp-3 gene expression 2h after challenge test, denoting that PHYTO diet reinforced fish capacity of stress response via protection of head kidney leucocytes from stress-related apoptotic processes, with lower caspase-3 gene expression and a higher il-1β gene expression when an infection occurs. Additionally, dietary supplementation with GMOS and PHYTO compounds increased fish serum lysozyme after infection. Both functional additives entailed a better capability of the animals to cope with infection in European sea bass when fed low FM and FO diets.

There is an increasing interest from the aquafeed industry in functional feeds containing selected additives that improve fish growth performance and health status. Functional feed additives include probiotics, prebiotics, organic acids, and phytogenics (substances derived from plants and their extracts). This study evaluated the effects of dietary inclusion of a mucilage extract rich in galactomannan oligosaccharides (GMOS), a mixture of garlic and labiatae-plants oils (PHYTO), and a combination of them (GMOSPHYTO), on gut microbiota composition of European sea bass (Dicentrarchus labrax) fed with a low fishmeal (FM) and fish oil (FO) diet. Three experimental diets and a control diet (plant-based formulation with 10% FM and 6% FO) were tested in a 63-days feeding trial. To analyze the microbiota associated to feeds and the intestinal autochthonous (mucosa-adhered) and allochthonous (transient) microbial communities, the Illumina MiSeq platform for sequencing of 16S rRNA gene and QIIME2 pipeline were used. Metabarcoding analysis of feed-associated bacteria showed that the microbial communities of control (CTRL) feed deeply differed from those of experimental diets. The number of reads was significantly lower in CTRL feed than in other feeds. The OTU (operational taxonomic unit) number was instead similar between the feeds, ranging from 42 to 50 OTUs. The variation of resident gut microbiota induced by diet was lower than the variation of transient intestinal microbiota, because feedstuffs are a major source of allochthonous bacteria, which can temporarily integrate into the gut transient microbiome. However, the composition of transient bacterial communities was not simply a mirror of feed-borne bacteria. Indeed, the microbial profile of feeds was different from both faecal and mucosa profiles. Our findings suggest that the dietary inclusion of GMOS (0.5%) and PHYTO (0.02%) in a low FM and FO diet induces changes in gut microbiota composition of European sea bass. However, if on allochthonous microbiota the combined inclusion of GMOS and PHYTO showed an antagonistic effect on bactericidal activity against Vibrionales, at mucosa level, only GMOSPHYTO diet increased the relative abundance of Bacteroidales, Lactobacillales, and Clostridiales resident bacterial orders. The main beneficial effects of GMOS and PHYTO on gut microbiota are the reduction of coliforms and Vibrionales bacteria, which include several potentially pathogenic species for fish, and the enrichment of gut microbiota composition with butyrate producer taxa. Therefore, these functional ingredients have a great potential to be used as health-promoting agents in the farming of European sea bass and other marine fish.

Aquaculture, the fastest growing food production sector cannot continue to rely on finite stocks of marine fish as the primary source of the omega-3 (n-3) long-chain polyunsaturated fatty acids (LC-PUFA), eicosapentaenoic acid (EPA; 20:5n3) and docosahexaenoic acid (DHA; 22:6n-3), for feeds. A four-month feeding trial was conducted to investigate the impact of a de novo oil, with high levels of EPA and DHA, obtained from transgenic Camelina sativa on growth performance, tissue fatty acid profiles, and expression of lipid metabolism genes when used as a replacement for fish oil in feed for European seabass (Dicentrachus labrax). Triplicate groups of 50 juvenile fish (initial weight 16.7 ± 0.92 g) per tank were fed for 4 months with one of three isolipidic and isoproteic experimental diets consisting of a standard diet containing a commercial blend of fish oil and rapeseed oil (CFO), a diet containing transgenic Camelina oil (TCO), or a blend of fish oil and rapeseed oil with enhanced levels of EPA and DHA (EFO) formulated to match the n-3 LC-PUFA profile of the TCO feed. Final weight of fish fed the GM-derived oil was not different to fish fed either CFO or EFO. Slight lower growth performance of fish fed TCO at the beginning of the trial was related to transient reduced feed intake, possibly caused by glucosinolates in the raw Camelina sativa oil. The GM-derived oil improved the nutritional quality of the fish fillet by enhancing total n-3 PUFA levels compared to the fish fed the other two feeds, and maintained flesh EPA and DHA at the same levels as in fish fed the diets containing fish oil. The metabolic response in liver and intestine was generally relatively mild although diets TCO and EFO seemed to trigger a metabolic response consisting of an up-regulation of both β-oxidation (cpt1a) and fatty acid transport (fabp1), possibly reflecting higher levels of LC-PUFA. Overall, the present study indicated that an oil of terrestrial origin, Camelina sativa, when engineered to contain high levels of EPA and DHA can replace fish oil in feeds for European seabass with no detrimental impact on growth or feed efficiency, while also maintaining or increasing tissue n-3 LC-PUFA contents.

This work brings together the view of different specialists in the areas of larviculture, physiology, ecology, nutrition and animal health, regarding how to deal with the aquaculture farming of species with complex life cycles in a multidisciplinary way, using as models the octopus and the muricid C. concholepas, with the aim of reducing the gap between the experimental and the industrial culture of species that are relevant for the diversification of aquaculture, particularly in Chile. Although these species are similar in their difficulty to reach the terminal planktonic phase prior to juvenile, they differ in the bottlenecks they have to overcome to reach it. Relevant aspects of study to achieve juvenile production from early life stages rearing, whether for repopulation or for ongrowing, are as follows: (i) Replacement or supplementation of live diets with inert diets to achieve significant survival values over the first stages of life. (ii) Physiological approaches to establish cultivation conditions evaluating the individual responses to several rearing conditions, specially the interaction between temperature, dissolved oxygen and acidity. Studies of urgent character due to the global warming scenario. (iii) Genomic studies associated with the effect of ontogenetic development, environment, health and nutrition on gene expression to understand, in an integrated way, the key processes for the development and growth of immature stages. (iv) Studies on the control of reproduction, the quality control of the ova, the genetic structure of reproductive populations and the characterization of diseases are also necessary to achieve efficient hatchery technologies., The International Workshop which enabled the discussionof these knowledge gaps was funded by INLARVI network of DID-UACh, Chile. Dr. Carlos Rosas also received fundingfrom the TEMPOXMAR network of UNAM, and DGAPAPAPIIT project IN219116, Mexico. Dr. Juan Carlos Navarroalso received funding from FONDECYT project 1131094.

The main objective of this study was to evaluate the stress response of the greater amberjack, Seriola dumerili, by determining plasma and skin mucus cortisol levels, as physiological indicator of fish stress, and skin mucin-2 gene expression as indicator of skin mucin production, under three different potential stressors related to aquaculture practices: temperature, handling and fasting. For trial 1, 45 greater amberjack juveniles (19.5 ± 4.1 g) were divided in three graded rearing temperatures (17, 22 and 26 °C) and maintained for 120 days, in order to define the basal cortisol level for this species after a long-term thermal acclimation within the range described for this species. Trial 2 was conducted under two different rearing densities and two different handling protocols with 222 fish (425.02 ± 36.9 g) divided in 4 treatments by triplicate for 90 days, in order to study the effect of routine handling protocols on the selected stress markers for this species. Trial 3 used 72 fish (302.27 ± 21.4 g) divided in two experimental groups, one fed to apparent satiety and another one subjected to fasting for 10 days (triplicates for each group). After trials 2 and 3, a shallow water challenge was conducted to further study the adaptive response of greater amberjack juveniles to combined stress situations. Plasma and skin mucus were obtained for cortisol determination at 1, 24 and 72 h (trial 2) and 1, 3 and 24 h (trial 3) post stress challenge. Additionally, skin was dissected out for determining muc-2 gene expression. Long-term acclimation to temperature did not affect plasma or mucus cortisol levels and muc-2 gene expression, being basal levels of circulating plasma cortisol around 7.5 ng/mL. A high stocking density (8 kg m−3) induced an elevation (p

Traditionally, a major part of aquaculture technology requires fish oil (FO) and fish meal (FM) to produce the aquafeed for farmed species. FO is the main source of persistent organic pollutants (POPs) in fish feed. In recent years, the use of vegetable-origin ingredients in fish feeds has been increasingly studied as an alternative to reduce the levels of these lipophilic pollutants in farmed species. The aim of this study was to evaluate the effect of the use of dietary vegetable oils in the farming of European sea bass (Dicentrarchus labrax) on the contents in persistent - polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) - and semi persistent pollutants - polycyclic aromatic hydrocarbons (PAHs) and polybrominated diphenyl ethers (BDEs) - of their edible parts. A total of 60 seabass muscle pools were obtained from fish farmed employing six experimental diets, which contained different percentages of FO (6 vs. 3%) and FM (20%, 10% and 5%). We did not observe differences in the contamination level of seabass muscle in relation to the percentage of FM in their diet. However, the fish farmed using feed which had lower levels of FO (3%) showed significantly lower muscle levels of ΣPCBs and carcinogenic PAHs (Σc-PAHs), with a reduction of 25.6% and 95.11% (respectively), as compared with those fished raised with feed with higher levels of FO (6%). Also much lower levels were found in OCPs such as sum of DDTs (30.88% of reduction), sum of chlordanes (42.85% of reduction), and sum of BDEs (48.16% of reduction) in those seabass fed with a lower percentage of FO. The results of this study indicate that the use of alternative feed ingredients that allow the employment of low percentage of FO in feeds help to reduce the load of several toxic pollutants in the fillets of European seabass.

The complete removal of fishmeal (FM) and fish oil (FO) is required to promote the sustainable development of aquaculture and for that, fast growing high quality fish that are fed without FM and FO are necessary. Early nutritional programming may allow the production of fish better adapted to utilize diets with vegetable meals (VM) and oils (VO). The main objective of this study was to research in the potential value of fatty acids as modulators of early nutritional programming in marine fish for a better utilization of VO/VM. For that purpose gilthead sea bream (Sparus aurata) broodstock were fed four different replacement levels of FO by linseed oil (LO) and their effect on fecundity and spawn quality, egg composition, Δ-6-desaturase (Δ6D) gene expression, progeny growth performance and their growth response to a challenge with diets low in FO and FM, but high in VO and VM. The results showed that feeding gilthead sea bream broodstock with high LO diets had very long-term effects on the progeny. Thus, FO replacement by LO up to 80–100% in broodstock diets for gilthead sea bream not only reduced fecundity and spawn quality, but also growth of 45 dah and 4-month-old juveniles, as well as Δ6D gene expression. However, when the 4 month-old juveniles were fed with a low FM and FO diet, even those from broodstock fed only 60% replacement of FO by LO showed a higher growth and feed utilization than juveniles from parents fed FO. These results demonstrate the interesting potential of early nutritional programming of marine fish by broodstock feeding to improve long-term performance of the progeny. Further studies are being conducted to determine optimum nutrient levels in the broodstock diets and the molecular mechanisms implied to develop effective nutritional intervention strategies for this species. Statement of relevance This study demonstrates for the first time in fish the potential of broodstock nutrition to conduct early nutritional programming of culture fish for a better utilization of low fish meal and fish oil diets by the progeny, showing its effect not only during reproduction and larval development but also during on-growing.

To date, European abalone aquaculture relies mostly on locally harvested fresh seaweeds which nutritional quality and abundance vary greatly, hence affecting abalone growth. Abalone artificial diets generally include fishmeal, limiting their utilization in ecologically sustainable aquaculture and affecting abalone quality and acceptance by the consumers. A six month feeding trial was conducted to assess the nutritional value of four different dried seaweed meals: Ulva lactuca (Chlorophyta), Gracilaria cornea (Rhodophyta), Laminaria digitata (Phaeophyta) and Palmaria palmata (Rhodophyta), as ingredients to all-vegetable-based formulated feeds for abalone Haliotis tuberculata coccinea (33.1 ± 0.8 mm and 4.7 ± 0.6 g). A mixed fresh algae diet of G. cornea and U. Ulva rigida, reared in an Integrated Multi-Trophic Aquaculture (IMTA) system, served as control. Survival rates were very high (95–98%), regardless of the diet fed. Enriched fresh algae produced a significantly higher growth for H. tuberculata coccinea (169% weight gain) than all the artificial diets (49–84% WG). Comparison among abalone fed the different formulated diets showed that the inclusion of P. palmata improved growth, condition index and dietary protein utilization. On the contrary, the use of L. digitata markedly reduced the efficiency of dietary protein since the protein-related nutritional index (PER), the percentage of protein deposited in the foot muscle as well as the meat to shell ratio recorded for animals fed this diet were the lowest, despite a higher feed intake. Large differences were found in the FA profile of fresh algae as compared with the three formulated diets. The n-3/n-6 ratio was much higher in the fresh algae and, consequently, in the foot tissues of abalone fed this diet in comparison to the one of those fed the formulated ones. The elevated contents of 20:4n-6 in the abalone fed the experimental diets and 20:5n-3 in abalone fed the fresh algae, as well as their respective metabolites, suggest that abalone have the ability to desaturate and elongate LA to ARA and ALA to EPA. Further studies are required to improve the growth obtained with these vegetable based diets, especially concerning the use of different seaweed combinations and inclusion levels, as well as the diet processing methods to improve diet water stability. Statement of relevance Development of vegetable based diets for abalone.

Despite the interest of meagre (Argyrosomus regius) as a fast-growing candidate for Mediterranean aquaculture diversification, there is a lack of information on nutrition along larval development. Importance of highly unsaturated fatty acids (HUFA) and the antioxidant vitamins E and vitamin C has not been investigated yet in this species. Six diets with two levels of HUFA (0.4% and 3% dw), two of vitamin E (1500 and 3000 mg kg−1) and two of vitamin C (1800 and 3600 mg kg−1) were fed to 15 dah meagre larvae. Larval growth in total length and dry body weight was significantly lowest in larvae fed diet 0.4/150/180 and showed few lipid droplets in enterocytes and hepatocytes and lower HUFA contents than the initial larvae. Increase in dietary HUFA up to 3%, significantly improved larval growth and lipid absorption and deposition. Besides, among fish fed 3% HUFA, increase in vitamin E and vitamin C significantly improved body weight, as well as total lipid, 22:6n-3 and n-3 fatty acids contents in the larvae. Thus, the results showed that 0.4% dietary HUFA is not enough to cover the essential fatty acid requirements of larval meagre and a high HUFA requirement in weaning diets is foreseen for this species. Besides, the results also pointed out the importance of dietary vitamin E and C to protect these essential fatty acids from oxidation, increase their contents in the larvae and promote growth, suggesting high vitamin E and C requirements in meagre larvae (higher than 1500 and 1800 mg kg−1 for vitamin E and vitamin C respectively).

The concept of nutritional programming raises the interesting possibility of directing specific metabolic pathways or functions in juvenile fish, for example, to improve the use of substitutes to fishmeal and oil, and hence to promote sustainability in aquaculture. The aim of the study was to determine effects of early nutritional stimuli of gilthead seabream larvae and check if nutritional programming of gilthead sea bream is possible between 16 days post hatching (dph) and 26 dph. A trial was conducted to determine the effects of early nutritional stimuli of gilthead seabream larvae. Five experimental microdiets (pellet size

Currently, the larviculture of many marine fish species with small-sized larvae depends for a short time after hatching, on the supply of high-quality live zooplankton to ensure high survival and growth rates. During the last few decades, the research community has made great efforts to develop artificial diets, which can completely substitute live prey. However, studies aimed at determining optimal levels of minerals in marine larvae compound feeds and the potential of novel delivery vectors for mineral acquisition has only very recently begun. Recently, the agro-food industry has developed several nano-delivery systems, which could be used for animal feed, too. Delivery through nano-encapsulation of minerals and feed additives would protect the bioactive molecules during feed manufacturing and fish feeding and allow an efficient acquisition of active substances into biological system. The idea is that dietary minerals in the form of nanoparticles may enter cells more easily than their larger counterparts enter and thus speed up their assimilation in fish. Accordingly, we evaluated the efficacy of early weaning diets fortified with organic, inorganic, or nanoparticle forms of trace minerals (Se, Zn, and Mn) in gilthead seabream (Sparus aurata) larvae. We tested four experimental diets: a trace mineral-deficient control diet, and three diets supplemented with different forms of trace minerals. At the end of the feeding trial, larvae growth performance and ossification, and the level of expression of six target genes (SLC11A2β, dmt1, BMP2, OC, SOD, GPX), were evaluated. Our data demonstrated that weaning diets supplemented with Mn, Se, and Zn in amino acid-chelated (organic) or nanoparticle form were more effective than diets supplemented with inorganic form of minerals to promote bone mineralization, and prevent skeletal anomalies in seabream larvae. Furthermore, nanometals markedly improved larval stress resistance in comparison to inorganic minerals and upregulated mRNA copy number of OC gene. The expression of this gene was strongly correlated with mineralization degree, thus confirming its potency as a good marker of bone mineralization in gilthead seabream larvae.

The longfin yellowtail ( Seriola rivoliana ) is a preferred species for aquaculture diversification in the Canary Islands. The aim of the present study was obtain a large number of spawns whit hormonal induction, which would provide reliable numbers of juveniles for the sustainable development of longfin yellowtail culture. Two females with oocytes bigger than 500 μ, and four males, previously captured from the wild were used for this study. Females and males of 10.7 ± 1.4 and 8.5 ± 0.9 kg body weight and 86.2 ± 5.3 and 78.9 ± 1.9 cm total length, respectively, were acclimated in 10 m 3 tanks. Fish were injected three times per month between June and November with 20 μg kg − 1 of GnRHa. During the experimental period, temperature ranged from 21.9 ± 0.2 °C in June to 23.5 ± 0.4 °C in November. Broodstock feeding consisted in Atlantic mackerel ( Scomber scombrus ) at 2% BW, three times per week. A total of 20,189,700 eggs (944,326 eggs female kg − 1 ) were obtained with fifteen injections and 33 spawns (2.2 ± 0.7 spawns per injection). A significant positive was found, between the numbers of eggs obtained each month and the average monthly temperature. Latency period was approximately of 28 h and 30 min. A highly significant negative statistical relation ( P − 1 of GnRHa every 10 days in fluent males and 500 μm oocytes bearing females of longfin yellowtail broodstock were successful to obtain a high induction efficiency and excellent spawn quality.

The present study aimed to determine the spawning efficacy, egg quality and quantity of captive breed meagre induced with a single gonadotrophin‐releasing hormone agonist (GnRHa) injection of 0, 1, 5, 10, 15, 20, 25, 30, 40 or 50 µg kg-1 to determine a recommended optimum dose to induce spawning. The doses 10, 15 and 20 μg kg-1 gave eggs with the highest quality (measured as: percentage of viability, floating, fertilisation and hatch) and quantity (measured as: total number of eggs, number of viable eggs, number of floating eggs, number of hatched larvae and number of larvae that reabsorbed the yolk sac). All egg quantity parameters were described by Gaussian regression analysis with R2=0.89 or R2=0.88. The Gaussian regression analysis identified that the optimal dose used was 15 μg kg-1. The regression analysis highlighted that this comprehensive study examined doses that ranged from low doses insufficient to stimulate a high spawning response (significantly lower egg quantities, p-1 through to high doses that stimulated the spawning of significantly lower egg quantities and eggs with significantly lower quality (egg viability). In addition, the latency period (time from hormone application to spawning) decreased with increasing doses to give a regression (R2=0.93) which suggests that higher doses accelerated oocyte development that in turn reduced egg quality and quantity. The identification of an optimal dose for the spawning of meagre, which has high aquaculture potential, represents an important advance for the Mediterranean aquaculture industry.

Haliotis tuberculata coccinea has been identified as a target species for European aquaculture development, in order to fulfil the rising demand for abalone. The effects of different stocking densities on the growth performance, feed utilization and survival of two different initial size groups (30 and 40 mm) of abalones, during the final grow-out to cocktail/market size (45–60 mm), were determined over a 6-month period. Trials were performed in abalone cages installed in a commercial open-sea cages fish farm. Animals were fed the red algae Gracilaria cornea and the green one Ulva rigida, both obtained from a land-based integrated multi-trophic aquaculture system. Survival rates were very high (94–98%) regardless the density employed. Sustained high linear growth was recorded both in shell and weight. However, a 17–19% reduction in weight gain was obtained by doubling the initial stocking density, suggesting a higher competition for space or food. Nevertheless, the high growth performance (70–94 lm day � 1 ; 250–372% weight gain) and survival attained, even at high densities, denoted the suitability of the offshore mariculture system as well as the biofilter produced macroalgae for grow-out culture of H. tuberculata coccinea that overall could reach cocktail/commercial size in only 18–22 months.

Reliable reproduction and sufficient amounts of juveniles obtained in captivity constitute a main bottleneck for mass production of greater amberjack. The aim of the present study was to combine several aspects of broodstock management and hormonal induction, to obtain a large number of high quality spawns. Captured Seriola dumerili sub-adults were kept for three years in 10 m3 tanks. When fish were larger than 75 cm, they were individually tagged, periodically explored to determine gonad maturation and injected with gonadotropin releasing hormone analogue from June to October. A total of 15 inductions to four males and two females produced 330 821 ± 219 519 eggs/induction and a total of 22 spawns. The number of spawns and female fecundity (2.48 millions eggs/female/spawning season) were similar to those of free wild populations and higher than those previously obtained in captivity for this species. Egg quality increased from an initial low in the first two spawns in June to give, during July to October, a mean of 96.01 ± 6.50% fertilized eggs, 92.58 ± 17.56% hatched eggs, 78.66 ± 12.60% larval survival at day 3 after. These results showed that intramuscular injections of 20 μg kg−1 body weight GnRHa under natural temperature and photoperiod conditions in the Canary Islands produced a high induction efficiency.

Escapes of farmed fish into the ecosystem have recently received a marked interest from scientists for their competition with wild populations or the risk of genetic contamination. Fatty acid profiles have been proposed as bio-indicators due to the inclusion of terrestrial oils in aquafeed ingredients. This study evaluated 1) the effect of wasted food on the fatty acid content of a farm-associated fish and 2) the suitability of fatty acid profiles as a bioindicator of aquaculture-ecosystem interactions, using the bogue ( Boops boops ) as a model. This species is an opportunistic fish usually associated with–or even found in–sea farms, and with a high natural occurrence along the Mediterranean and Atlantic coasts. The results showed that fish farms had a direct effect on the condition index, muscle and whole lipid content of bogue inside or around the sea cage, and this effect disappeared completely at 3 km from the cages. Despite the fact that aquafeeds also affected the bogue fatty acid profiles by increasing linoleic and oleic acids and reducing DHA, these profiles were very similar to those of bogue sampled close to a sewage outfall, denoting that these fatty acids are not exclusively influenced by aquaculture and therefore should not be considered as good biomarkers for escapees. However, bogues influenced by aquaculture were higher in linolenic acid than those found further than 3 km from the farm or close to the sewage, suggesting this fatty acid as a better indicator of aquaculture influence.

Despite considerable progress in recent years, many questions regarding fish larval nutrition remain largely unanswered, and several research avenues remain open. A holistic understanding of the supply line of nutrients is important for developing diets for use in larval culture and for the adaptation of rearing conditions that meet the larval requirements for the optimal presentation of food organisms and/or microdiets. The aim of the present review is to revise the state of the art and to pinpoint the gaps in knowledge regarding larval nutritional requirements, the nutritional value of live feeds and challenges and opportunities in the development of formulated larval diets.

Dietary lipid source in aquaculture has become a central topic in research whilst natural resources availability diminishes. Hence, to weigh up and forecast consumers impressions, the impact of partial (70%) and complete (100%) dietary replacement of fish oil (FO) by linseed oil (LO) on sensory and quality attributes was studied during the edible shelf life of gilthead sea bream (Sparus aurata). Physico-chemical parameters (pH, torrymeter, total volatile basic nitrogen, thiobarbituric acid reactive substances and texture), and sensory analysis, both in cooked and raw fish were carried out during 17 days of ice storage. Throughout ice storage, feeding with LO diets, TBARS values remained lower on muscle than those found when feeding FO control diet. On freshly caught fish (day 0 of ice storage), statistically significant dietary texture variations were recorded on cooked fillet fed FO diet. No sensory differences on Quality Index Method, sensory profile or Torry scheme were found with partial or total LO replacement diets.

Octopus vulgaris is a potential candidate to diversify marine aquaculture. Even though mortality under rearing conditions has been related to reproductive processes, the benefit of sex segregation remains unclear. In order to clarify this, wild octopuses (862 ± 101 g) were reared in floating cages under three proportions of male : female sex ratio: 1:1, 1:0, and 4:1 (n = 30 per treatment). The experimental period lasted 2 mo and octopuses were fed on bogue, Boops boops (aquaculture by-product). Higher growth rates were observed in octopuses reared under 1:1 and 1:0 (1.8%/d) in comparison with those reared under 4:1 conditions (1.6%/d). Regarding sexes, a lower growth was detected in females reared under 4:1 (1.1%/d) in comparison with females (1.8%/d) and males (1.7–1.9%/d) reared under 1:1 conditions. Survival was 97, 97, and 90% in sex ratio 1:1, 1:0, and 4:1, respectively. Sexual maturity data showed that males were all mature, while most females were still maturing (60–84%) at the end of the rearing period. High lipid content in bogue (44% dw) did not reflect on octopus muscle. This tissue showed a similar biochemical composition irrespective of the sex ratio condition, with a high protein (87% dw), a low lipid (5% dw), and a high n-3 HUFA content (42%).

Two fresh diets were tested on the biological performance of males and females of Octopus vulgaris under industrial rearing conditions in floating cages. The presence of females could potentially release paralarvae to the natural environment (‘integrated aquaculture’). Octopuses were fed on a single diet of bogue (aquaculture by-product), and on a mixed diet (60–40% crab-bogue), in two trials. In Trial 1 (918 ± 125 g), growth was higher in octopuses fed on the mixed diet (1.9–2.0%d−1) than in those fed on the bogue diet (1.8–1.9%d−1) irrespective of sex, and mortality was 3% regardless of sex and diet. In Trial 2 (1483 ± 269 g), growth was higher in males fed on the mixed diet (1.8%d−1) than in males fed on the bogue diet (1.4%d−1), both higher than females (1.1–1.3%d−1), and mortality was 22–28% regardless of sex and diet. Best biological performance in trial 1 was related to the lack of functional maturation in females, evidence by the low average gonadosomatic index (2.0–3.6%) in comparison with trial 2 (8.8–11.4%), regardless of diet. No correlations between final weight and sexual maturity indices were found, so the length of the rearing period to maximize biomass increment or to release paralarvae depends on functional maturation in females.

The present study describes the adaptation of longfin yellowtail Seriola rivoliana as broodstock and first larval rearing trials under intensive and semi-intensive conditions. Fifteen sub-adults were captured in the South coast of Gran Canaria (Canary Islands, Spain) in June 2007. Fish (initial weight 1.76 ± 0.25 kg) reached a weight of 6.0 ± 1.1 kg in July 2010. Once a year, fish were sampled to determine individual growth in weight and size. In addition, the state of sexual maturity was established based on gonadal biopsies. On the basis of repeated hormonal injection (GnRHa, 20 μg/kg), 10 successful spawns were obtained between July and October 2009, with 92.5 ± 5.5% and 72.6 ± 17.2%, fertilization and egg viability respectively. First results of larval rearing under semi-intensive conditions, showed an average survival at 30 DAH of 2.5% as compared with 0.5% under intensive conditions. The low survivals under the two rearing conditions in addition to their failure to pass a stress test could be attributed to deficiencies in essential fatty acids as could be seen in both eggs and feeds. Morphometric parameters showed no significant difference between the two rearing systems in 230 DAH larvae.

Some benthic cephalopods are considered potential candidates to diversify marine aquaculture, as they show fast growth and high market price. Most research on cephalopod culture is currently focusing on the development of specific enrichments and compound feeds, while little research has been conducted in order to test new rearing systems for cephalopods. The rigid characteristic of the floating cages commonly used for the ongrowing of Octopus vulgaris has restricted their use to calm water conditions (estuaries and harbors). Such sites are scarce and highly demanded, especially by the tourism industry; therefore the development of O. vulgaris grow out at these locations competes with touristic interests. The present study was set to compare the biological performance of O. vulgaris reared in a benthic cage (2 m 2 ) as opposed to the traditional floating cage (2.5 m 2 ), during two ongrowing trials. Initial rearing density was 10 kg m −3 and octopuses (892 ± 125 g) were fed on bogue Boops boops , discarded from fish farms, for 60–67 days. High growth (1.8–1.9% day −1 ) and high survival (91–97%) were observed, regardless of the rearing system, and led to best biomass increment (178–212%) and food conversion rates (2.3–2.6) ever recorded for O. vulgaris under industrial rearing conditions. These results underline the adequacy of the benthic cage for the ongrowing of this species, and also the potential of aquaculture discarded fishes, particularly bogue, as a single diet for this species. High growth rates obtained and the high lipid content of bogue (44% dry weight) suggest efficient lipid utilization in this species. Proximate composition and fatty acid profile in octopus muscle was not affected by the rearing system. High dietary lipid content was not reflected in muscle proximate composition, which showed high protein (87% dw) and low lipid content (5% dw) by the end of the experimental period. Farmed octopus showed high levels of n -3 HUFA (42%), which should enhance its value for the consumers.

The octopus, Octopus vulgaris , is one of the main targets for aquaculture diversification in Mediterranean countries. However, the development of octopus farming is limited by the lack of information regarding nutritional requirements of this species during its life cycle. In this study, five diets were tested on the biological performance (growth, protein retention and biochemical composition) of individually reared octopuses (n = 8 per diet), including three single diets constituted by: an endemic crab (the white crab, Plagusia depressa ), a commercial crab imported frozen (the blue crab, Portunus pelagicus ), and bogue ( Boops boops ) discarded from fish farms (aquaculture by-product), as well as two mixed diets, containing a 60–40% of blue crab-bogue and white crab-bogue, respectively. The rearing period lasted 8 weeks. Octopuses that fed on a mixed diet constituted by blue crab-bogue showed a higher growth than those feeding on bogue as a single food item. No significant differences in growth were observed among individuals feeding on single food items. Highest protein retention was observed in octopuses fed on diets containing discarded bogue, associated with a high lipid and monoenes content in this food item, underlying the use of lipid as energy source in O. vulgaris . However, discarded bogue was deficient in ARA in comparison with octopus tissues, which did not seem to affect growth during the experimental period. These findings underline the potential of aquaculture by-products, particularly bogue, as an adequate diet for culturing O. vulgaris .

Abalone Haliotis tuberculata coccinea Reeve (1846), is a target species for diversification of European aquaculture production. Taking into account that sustainable, eco-friendly production methods are to be a part of future expansion of the abalone industry, growth performance of juvenile abalone reared in an integrated culture system was evaluated and compared with that of abalone fed non-enriched macroalgae. Four macroalgae treatments, three monospecific: Ulva rigida (UN), Hypnea spinella (HN) and Gracilaria cornea (GN) and a composite one (MN), were produced out of fishpond wastewater effluents, while other four control treatments consisted of the same species reared in fresh seawater (U; H; G; M). Seaweeds reared in fishpond wastewater effluents increased their protein content from 11–17% to 29–34%. Lipids consisted mainly of saturated fatty acids (SFA) (43–60%), palmitic acid being the most abundant fatty acid (40–47%). Highest EPA percentage was found in red algae H. spinella (6.9%), being ten times higher than that of U. rigida (0.7%). All the algae tested contained very low levels of arachinodic acid (0.1–1.6%) and docosahexaenoic acid (0.5–3%). Protein levels in foot muscle (74–76%) did not differ significantly (P G. cornea led to the lowest growth performance probably due to the lowest feed intake. The results clearly indicate that H. tuberculata coccinea growout can efficiently take place in an integrated-culture system suggesting that on-farm seaweed-abalone production could be part of future development of the abalone industry in the Canary Islands.

The physiological response to stressors in fish, including hormonal profiles and associated tissue responsiveness, is less documented. The aim of this study was to evaluate feeding gilthead sea bream ( Sparus aurata ) with diets containing linseed oil (LO) and soybean oil (SO) as substitutes to fish oil (FO) and their effect on fatty acid profile of head kidney and the consequent effect on stress response to a crowding challenge. Fish were fed 8 experimental diets with different levels of substitution 0% (FO), 70% (70LO, 70SO, 20LO50SO and 50LO20SO) and 100% (100LO, 100SO and 50LO50SO) over a period of 8 months. At the end of the feeding trial, samples of head kidney were collected for biochemical analysis and the fish were challenged by a crowding test. During the challenge, samples of plasma for cortisol analysis were collected at 0 h, 2 h, 5 h, 24 h, 48 h and 1 week in order to study acute and chronic stress responses. Results showed that fish fed vegetable oils (VO) had significantly decreased ARA, EPA, DHA and n-3 HUFA, while LA, LNA and total C18 PUFAs were significantly increased. The basal cortisol levels were significantly increased in fish fed 70LO, 100LO, 50LO20SO and 50LO50SO. The physiological response to crowding was significantly affected by the diet. After 2 h of crowding, all the treatments showed higher cortisol, with fish fed 100LO had significantly the highest response registering 131.38 pg/ml. After 5 h and 24 h, plasma cortisol was reduced in all treatments except in 50LO20SO. After 48 h of crowding, the plasma cortisol was increased in all treatments with the maximum value seen in fish fed 100LO (72.12 pg/ml). These levels were decreased in fish fed FO, 70LO, 100LO and 50L050SO after 1 week of crowding, but remained higher in fish fed 70SO, 100SO, 20LO50SO and 50LO20SO. In conclusion, fish fed LO diets showed the same response pattern as the control but with higher intensity regaining the basal levels after 1 week as the control, while fish fed SO had a slow response but changed the pattern characterized by a lower response at the beginning and longer recuperation without regaining the control value even after 1 week.

Octopus vulgaris is a species of demand in the market with the potential to diversify European aquaculture. However, this species develops complex social interactions under culture conditions, which may have negative effects on its growth, survival and profitability. In order to understand its behaviour under such conditions, individual tagging systems allow a careful evaluation of biological parameters, such as growth and longevity. The present work describes a combined protocol (anaesthetic and tagging) for implanting subcutaneous passive integrated transponder tags (PIT). The effect of two anaesthetic agents in facilitating octopus handling is assessed: clove oil at 20–40–100 mg L−1 and ethanol (96%) at 1–1.5–2%. The most suitable body location of PIT tags, its effect on growth and mortality, the addition of a stitch and the PIT retention rate after 2 months in floating cages were evaluated. It was concluded that immersion in seawater with 1.5% of ethanol at 22.3±0.5 °C is a suitable anaesthetic for this species. The results showed that the best-selected PIT body location was the upper left arm III. No effect of the PIT tagging system was found on growth and survival when tagged and untagged octopuses were compared. It was observed after 2 months that the stitch did not induce an increase in the retention rate and 81–100% tag retention regardless of the dietary treatment.

Estefanell, J., Socorro, J., Roo, F. J., Fernández-Palacios, H., and Izquierdo, M. 2010. Gonad maturation in Octopus vulgaris during ongrowing, under different conditions of sex ratio. – ICES Journal of Marine Science, 67: 1487–1493. Octopus vulgaris is a suitable candidate for aquaculture, but there are problems with breeding in captivity, such as aggressive behaviour among males and the frequent death of females after the eggs hatch. To avoid these problems and further understand the sexual maturation of common octopus in captivity, males and females were reared together and separately under similar culture conditions. In all trials, the initial rearing density was 10 kg m−3. Females (n = 15, sex ratio 0:1) and males (n = 11, sex ratio 1:0) were kept in circular tanks, and a mixed group (n = 209, sex ratio 4:1) in floating cages. Trials started in November 2008 and octopuses from each treatment were examined macroscopically and histologically in December and January to assess sexual maturation. All the males matured, regardless of the sex ratio during rearing, as did all females in the mixed group. In contrast, a large proportion of the females kept isolated from males was still immature in December and January. Although maturation was successful in floating cages, there was 76% mortality there, in contrast to the zero mortality in tanks. Moreover, most of the dead octopuses from the cages were in post-reproductive condition, with a low digestive gland index, suggesting that this was natural post-reproductive mortality. Therefore, sex segregation is deemed advantageous to avoiding early mortality.

The present study describes the osteological development and the occurrence of skeletal deformities in red porgy Pagrus pagrus larvae in relation to the intensification of the rearing system. Eggs obtained from natural spawning were cultured under two different rearing systems: intensive (100 eggs l⁻¹) in 2000 l and semi-intensive (mesocosm) system (5 eggs l⁻¹) in 40,000 l conico-cylindrical tanks. Fish samples were periodically collected along the development from hatching to juveniles at 95 days post hatching (dph). Osteological development, meristic counts and the presence of skeletal deformities were evaluated. Despite the external appearance of the juveniles being similar to wild standards, X-ray studies revealed a high number of fish (semi-intensive: 37·8%; intensive: 45·5%) with skeletal deformities. Regardless of the rearing system, no significant interaction was found between the per cent of the most common deformities, axial deviations (lordosis and presence of fused vertebrae). Cranial deformities and kyphosis incidences, however, were significantly higher in intensively cultured P. pagrus. Also, the fused vertebrae in these fish were located mainly in the caudal area instead of pre-haemal area for semi-intensively reared P. pagrus. Moreover, a significant interaction was found between the total number of vertebrae and the type of rearing system used; fish from the intensive system showing a higher number of fish with an extra vertebrae (10 abdominal + 15 caudal). Present results suggest a relationship among feeding sequence, osteological development and deformity incidence and location in P. pagrus larvae.

Despite the importance of certain highly unsaturated fatty acids in osmotic regulation, few studies have been addressed to determine the essential fatty acid requirements for a given species cultured under different salinities. As Galaxias maculatus is a diadromic species, the present study aimed to determine the effect of salinity on the optimum dietary EPA/docosahexaenoic (DHA) ratio for survival and growth during the larval stages. Larvae were fed for 20 days with rotifers containing two different EPA/DHA ratios (low: 0.64 and high: 2.18) at three different salinities (0, 10 and 15 g L−1). The results of this study showed a marked effect of water salinity on larval dietary lipid utilization in G. maculatus larvae. These results suggested that G. maculatus larvae reared at higher salinities may have a higher dietary requirement for DHA, whereas larvae reared at 0‰ showed higher requirements for EPA. The overall results of the present study indicate that even small changes in salinity can determine the optimum dietary EPA/DHA ratio and the quantitative essential fatty requirements of fish. This may have important repercussions and affect the rearing performance of G. maculatus cultured under different salinities.

Summary Despite the great interest of red porgy as a new species for Mediterranean aquaculture, its commercial production is constrained by the high incidence of skeletal deformities occurring in this species under culture conditions. Several studies have been conducted to better understand the origin of these anomalies in this species, using different system intensiveness, rotifers enrichment products or rotifers docosahexaenoic acid content. The first study showed that culture intensification increased the number of fish with an extra vertebrae, what was probably related to the different nutritional quality of live preys employed in each treatment, since water temperature, salinity and genetic background were identical for the different batches of fish studied. Total incidence of skeletal abnormalities was higher in the intensive system, particularly cranial abnormalities and kyphosis in the cephalic vertebrae. In both rearing systems the most common skeletal anomalies were vertebral column disorders, lordosis and fused vertebrae, their localization along the column being affected by the culture intensiveness. Rotifer enrichment, predominantly its docosahexaenoic acid content significantly affected deformities occurrence. A marked positive effect of rotifer docosahexaenoic acid content was found on larval survival. X-ray studies denoted elevated levels of bone abnormalities associated, in both trials, to low docosahexaenoic acid content in live preys. Among different anomalies, the presence of fused vertebrae was the most frequent deformity for both rearing trials. A 50% reduction in the number of deformed fish for each type of deformity was obtained when the larvae were fed higher docosahexaenoic acid levels, denoting the important role of this fatty acid in bone development. Further studies are needed to elucidate the importance of essential fatty acids on the development of bone deformities in fish, since the functions of these fatty acids differ among them and can lead to very different effects in fish metabolism, including bone formation.

Summary Red porgy is a candidate species for marine aquaculture diversification. The objective of the present study was to describe the osteological development in this species and the occurrence of skeletal deformities in relation to the intensification of the rearing system. Fish samples were periodically collected along the development from hatching to juveniles (95 days after hatching). Osteological development and the presence of skeleton abnormalities were evaluated. Larvae reared under S-IS showed a better growth in terms of total length in comparison with IS reared ones. Regarding to osteological development for red porgy, this was similar between fish from both culture systems, but differing in timing of apparition and ossification of skeletal elements. X-ray studies revealed a high number of fish with skeletal deformities (Semi-intensive: 38.8%; Intensive: 46.5%), but no significant effect of the rearing technique on the incidence of deformities such as lordosis or fused vertebrae was found. However, cranial abnormalities and kyphosis incidences were significantly higher in intensive system cultured red porgy. These results, suggest a relationship among rearing technique, osteological development and the apparition of certain deformities.

The promotion of nutritional strategies that optimize natural defence mechanisms in fish is of critical importance in producing robust juveniles and adult fish. These animals are more capable of minimizing the impact of opportunistic pathogen attack, thus improving liveability and performance characteristics. The importance of the piscine gastrointestinal tract as a major endocrine and osmoregulatory organ is well reported as is its function as a defensive barrier to pathogen attack. Investigations using the inclusion of a specific structural form of mannan oligosaccharide have been shown to improve the performance parameters, immune status, and gut morphology and improve an important aspect of barrier protection through the enhancement of mucal production in a number of aquaculture species. The selenium status of an animal is pivotal in determining the success of the innate and adaptive immune response of the animal, and the use of an organic selenium source, in the form of a selenoyeast, has been shown to improve enzyme function and tissue uptake. The antioxidant role of many of the selenoptroteins and the role of selenium in the glutathione peroxidase enzyme pathways involved in the control of oxidative stress is critical if oxidative damage and cell membrane lipid peroxidation are to be prevented. The use of these compounds as feed additives has important implications for health management in commercial aquaculture facilities. Further research is needed to evaluate the benefits offered by a range of commercial products.

Despite being proposed as a potential candidate for diversification of marine aquaculture, limited larval survival and the occurrence of elevated levels of skeletal deformities restrict the commercial production of red porgy. The present study was conducted to determine the effect of rotifer enrichment, particularly on DHA, on growth, survival and occurrence of skeleton deformities in this species. The study included two trials, in one the viability of commercial emulsions for rotifer enrichment was evaluated whereas in another the effect of the elevation of DHA in rotifer emulsions on the larvae was studied. No significant differences were found in growth between larvae fed different rotifers in both trials suggesting that 1.9% dw DHA is enough to fulfil the DHA requirements of red porgy larvae for maximum growth. However, a marked positive effect of rotifer DHA content supplementation on larval survival was found. A low larval survival was found when rotifers had a high DPA content, in agreement with the results obtained for other Sparids. This fatty acid was accumulated in red porgy larval tissues in high levels (0.79% dw DPA) when rotifers rich in DPA were the main source of food. Although the external appearance of the juveniles seemed to be normal, X-ray observations showed elevated levels of bone abnormalities associated, in both trials, to low DHA content in the live prey. Among different anomalies, the presence of fused vertebrae was the most frequent deformity for both rearing trials. Although DHA content in the rotifers used for feeding was the only difference a 50% reduction in the number of deformed fish for each type of deformity studied was obtained when the larvae fed higher DHA levels, denoting the important role of this FA in the prevention of deformities at the rotifer feeding stage. Further studies are needed to elucidate the importance of essential fatty acids on the development of bone deformities in fish, since the functions of HUFA are different, and their absolute levels and ratios among them can lead to very different effects in fish metabolism, including bone formation.

Red porgy, Pagrus pagrus, is a candidate for aquaculture diversification in the Mediterranean area. The main restriction for the development of this species, under cultured conditions, is the loss of their natural red–pink coloured skin. Several studies, using different dietary astaxanthin sources have given positive results on red porgy skin coloration. However, there is scarce information on the time required for cultured red porgy to obtain adequate skin colour when astaxanthin is included in the diet. This study aims to evaluate the effect of supplementing adult red porgy diet with shrimp shell meal, as a source of esterified astaxanthin, during different periods prior to harvest, on skin colour and skin carotenoid concentration. The experiment was carried out for 180 days, testing a control diet with no carotenoids and a shrimp shell meal diet (SM) with an inclusion of 16% shrimp shell meal in substitution of fish meal. Four treatment groups were established. Control treatment fish fed throughout the experiment on the control diet and SM60, SM120 and SM180 treatment fish fed first on the control diet and thereafter for 60, 120 and 180 days before harvesting on the SM diet. Growth was higher in fish from SM180 treatment group in comparison to control fish. Skin redness (a⁎) increased with increasing feeding period, and skin yellowness (b⁎) only until 120 days of feeding with SM diet. Whereas skin lightness (L⁎) does not seem to be influenced by dietary inclusion of shrimp shell meal for different periods of time. Skin carotenoid concentration also augmented with increasing feeding with SM diet. As a result, cultured red porgy fed for a period of 120 to 180 days, prior to harvest, on the SM diet may achieve similar skin coloration to wild red porgy, only differing in skin carotenoid concentration.

This study examines the utility of Passive Integrated Transponder (PIT) tagging system in fingerlings of gilthead seabream, Sparus auratus L. Two different body positions (abdominal, dorsal) and four weight classes (2–3 g, 3–4 g, 4–5 g, 5–8 g) were tested in three experiments. Tag loss rate, as well as its effect on growth and mortality rates, was determined. Times of handling and healing were also assessed. Fish tagged in the abdominal cavity showed a significantly lower tag loss rate (14%) than those given dorsal muscle implants (40%). No differences were found in growth within experiment between tagged and untagged fish, their final mean weight ranging between 6–39 g and 6–38 g, respectively. There was no difference in mortality between tagged and untagged fish at any position (0–3.4% and 0–2.1%, respectively). Fish smaller than 3 g suffered significantly higher mortality (14.3%), indicating that the PIT tags are not suitable for very small individuals. PIT retention rate was 100% above 4 g, irrespective of the personal expertise in tagging. Mean tag application time was 19 s per fish. Mean wound healing time was close to 20 days. Hence, these results showed that PIT tagging of gilthead seabream above 3 g is feasible because it does not affect growth or mortality.

In the present study 75 g European sea bass were fed for 8 months with different diets (22% dietary lipid content) containing vegetable oils at two inclusion levels. The control diet (Diet FO) contained anchovy oil as the only lipid source; in diets 60RO, 60LO, and 60SO of fish oil was substituted by rapeseed, linseed or soybean oils, respectively; finally, in diet 80LO, 80% of the fish oil was substituted by linseed oil. Fish were fed to apparent satiation three times a day. All fish were individually weighed once per month. Lipid and fatty acid composition of diets and fish fillets were determined at the beginning, middle and end of the experimental period. Once the commercial size was reached, all fish were fed a 100% FO containing diet during 150 days. No significant differences were found in feed intake. Fish fed the diets containing 80% linseed oil or 60% rapeseed oil had significant ( P After 150 days of re-feeding period with a 100% fish oil diet, DHA levels were restored in those fish previously fed diets containing vegetable oils, but EPA levels remained lower when compared with fish fed 100% FO diet. Flesh content of linoleic and linolenic acids remained higher in those fish previously fed soybean and linseed oil containing diets, respectively.

A 60-day feeding trial was conducted to assess the suitability of three red algae, Hynea spinella, Hynea musciformis and Gracilaria cornea, as potential feed for the culture of juvenile abalone, Haliotis tuberculata coccinea R. Seaweeds were reared in a biofiltration unit with fishpond waste water effluents. The three algal species were found to contain high protein contents which would be related to its production under the high nitrogen culture conditions of the biofilter system. Protein and carbohydrate contents were highest in H. musciformis and lowest in G. cornea. Survival rates of juvenile abalone were very good, regardless of the algae fed. Feed intake of H. spinella was highest, followed by H. musciformis. Growth rates of abalone were within the range obtained under commercial conditions, final shell length and weight being significantly highest in animals fed H. spinella and lowest in those fed G. cornea. Feeding G. cornea lead to the lowest growth performance due to the lowest feed intake, whereas feed conversion ratios were significantly highest for H. musciformis and protein efficiency ratios were higher for both H. spinella and G. cornea. This study suggested the good potential of any of the three red seaweeds tested—successfully produced by the biofilter system, their nutritional composition being similar to other macroalgae used as feed for abalone and matching the abalone protein and lipid requirements—hence promoting growth and survival. Nevertheless, the biofilter produced macroalgae H. spinella showed the highest dietary value for juvenile of H. tuberculata coccinea.

This study investigates the effect of dietary vitamin E on juveniles of gilthead seabream under stressful situations, focusing on the effects on growth, haematology, some immune parameters and plasma cortisol as indicators of stress. Two sardine meal-based experimental diets, one of them supplemented with 150 mg of alpha tocopherol kg(-1) of diet (control) and another one without vitamin E supplementation (diet NE), were assayed under two different stress conditions: overcrowding as a chronic stressor (during 15 weeks) and repetitive chasing as an acute repetitive stressor. Low levels of vitamin E in the diet depleted alternative complement pathway activity [from 167.23 U ml(-1) (control fish) down to 100.99 U ml(-1)] and also nonspecific haemagglutination. Also, fish fed a non-supplemented diet showed an elevation of plasma cortisol basal levels without a stressor influence [from 3.91 ng cortisol ml(-1) plasma (control fish) up to 21.70 ng cortisol ml(-1) plasma]. Low levels of vitamin E in the diet also produced an increase of erythrocyte fragility. Under chronic stress, fish fed the vitamin E-deficient diet showed a reduction in growth and survival, and alterations in haematological parameters, such as an additional haemoconcentration in response to overcrowding when compared with control fish. Under repetitive stress, fish fed the vitamin E deficient diet showed faster elevation of plasma cortisol levels in response to stress and a lower survival rate than control fish. Production of oxygen radicals by blood neutrophils was reduced under repetitive stress in fish fed the non-supplemented diet. These results suggest that fish fed the vitamin E-deficient diet had lower stress resistance.