Towards an inert diet for first-feeding gilthead seabream Sparus aurata L. larvae

The development of an inert food to replace live prey during the early stages of marine fish larvae requires research in different fields and therefore a precise work strategy. Our research on this subject has been carried out in successive steps using the gilthead seabream Sparus aurata . The first step was the design of a food particle that would be well accepted and ingested by free-swimming marine larval fish during the first developmental stages. We chose microencapsulation by polymerization of the dietary protein as the most appropriate method for making the particles; different types of microcapsules were made using a basic diet containing only the major dietary components. In the second step, our aim was to keep the larvae alive in a routine rearing system in 300-L tanks, using exclusively this kind of food, long enough to detect any changes in growth, survival, or anatomical and histological status of the larvae, in order to verify whether the technological changes were positive. The third step focused on diet formulation and searching for clues to inefficient assimilation and growth. The use of ' in vitro ' digestibility techniques allowed us to detect the inhibitory effect of some diet ingredients on larval proteases and to determine more suitable sources of protein. We now have a microcapsule able to efficiently support growth and development of S. aurata larvae , at least during the first 2 weeks of life, although the larvae still need to feed on rotifers during the first 2–4 days of exogenous feeding. This microcapsule will make it possible to make advances in determining the specific nutritional requirements of larval fish.     

Optimum soybean lecithin contents in microdiets for gilthead seabream (Sparus aurata) larvae

The aim of the present study was to determine the optimum dietary levels of soybean lecithin (SBL) for seabream (Sparus aurata) larvae, and its influence on production performance and digestive enzymes activity. Larvae were fed five formulated microdiets with five levels of SBL. Complete replacement of live preys with the experimental microdiets for seabream larvae at 16 dph produced over 55% survival rates, particularly in fish fed with the highest levels of SBL. Moreover, increase in dietary SBL up to 80 g kg⁻¹ significantly improved larval growth, leading to high final total length and body weight. An increase in alkaline phosphatase activity with the elevation up to 80 g kg⁻¹ SBL was also found denoting a better maturation of the digestive system. Besides, there was a stimulatory effect of dietary SBL on PLA2 activity. Finally, increasing dietary SBL lead to better utilization of dietary highly unsaturated fatty acid, as it was reflected in their higher content in both neutral and polar lipid of the larvae. In summary, elevation of dietary SBL up to 80 g kg⁻¹ in microdiets for seabream significantly improved digestive enzymes activities, enterocyte maturation, utilization and deposition of dietary essential fatty acids and larval growth, as a consequence of a better digestion, absorption, transport and deposition of dietary nutrients.     

Optimum krill phospholipids content in microdiets for gilthead seabream (Sparus aurata) larvae

The aim of the present study was to determine the optimum dietary levels of krill phospholipids (KPL) for sea bream (Sparus aurata) larvae, and its influence on larval development and digestive enzymes activity. Larvae were fed five formulated microdiets with five different levels of KPL. Complete replacement of live preys with the experimental microdiets for seabream larvae produced high survival and growth rates, particularly in fish fed the highest levels of KPL. In the present study, increase in dietary KPL up to 120 g kg^?1 (100 g kg^?1 total PL) significantly improved larval survival and growth, whereas further increase did not improve those parameters. An increase in alkaline phosphatase, trypsin and lipase activity with the elevation of KPL up to 120 g kg^?1 was also found denoting a better functioning of digestive system. Besides, there was a linear substrate stimulatory effect of dietary KPL on phospholipase A2 activity. Finally, increasing dietary KPL lead to better assimilation of n‐3 HUFA especially eicosapentaenoic acid, reflected in the higher content of these fatty acids in both neutral and polar lipids of the larvae. In summary, KPL were found to be an excellent source of lipids for seabream larvae. Optimum inclusion levels of this ingredient in microdiets to completely substitute live preys at this larval age were found to be 120 g kg^?1 KPL.     

Transmission of lymphocystis disease virus to cultured gilthead seabream,Sparus aurata L., larvae

The transmission of lymphocystis disease virus (LCDV) to gilthead seabream, Sparus aurata L., larvae was investigated using fertilized eggs from a farm with previous reports of lymphocystis disease. LCDV genome was detected by PCR‐hybridization in blood samples from 17.5% of the asymptomatic gilthead seabream broodstock analysed. Using the same methodology, eggs spawned from these animals were LCDV positive, as well as larvae hatched from them. The presence of infective viral particles was confirmed by cytopathic effects development on SAF‐1 cells. Whole‐mount in situ hybridization (ISH) and immunohistochemistry (IHC) showed the presence of LCDV in the epidermis of larvae hatched from LCDV‐positive eggs. When fertilized eggs were disinfected with iodine, no viral DNA was detected either in eggs (analysed by PCR‐hybridization) or in larvae (PCR‐hybridization and ISH). These results suggest the vertical transmission of LCDV, the virus being transmitted on the egg surface. Larvae hatched from disinfected eggs remain LCDV negative during the endotrophic phase, as showed by PCR‐hybridization, ISH and IHC. After feeding on LCDV‐positive rotifers, viral antigens were observed in the digestive tract, which suggests that viral entry could be achieved via the alimentary canal, and that rotifers can act as a vector in LCDV transmission to gilthead seabream larvae.     

Eggs of gilthead seabream Sparus aurata L. as a potential enrichment product of Brachionus sp. in the larval rearing of gilthead seabream Sparus aurata L.

The gilthead seabream Sparus aurata L. is a sequential spawner whose reproductive cycle can be manipulated to obtain year-round egg and fry production. Hence, there is an overproduction of eggs in commercial hatcheries that is normally wasted. Two feeding experiments were conducted with the purpose of investigating the potential use of fresh and freeze-dried gilthead seabream eggs for the enrichment of live preys. The first experiment was also designed to study the possibility of using freeze-drying as a conservation process for eggs in excess. Larvae were reared for 20 days after hatching (DAH),on a diet of Brachionus sp. As experimental controls, the commercial enrichment emulsion SUPER HUFA was used in the first experiment, and the microalgae Nannochloropsis sp. in the second. This study demonstrated the potential of gilthead seabream eggs as an enrichment product to be used in the feeding of the first larval stages of Sparus aurata L. The data obtained emphasize the need to have an overall perspective of the nutritional requirements of the organisms in culture.     

The effect of dietary exogenous digestive enzymes on ingestion,assimilation, growth and survival of gilthead seabream (Sparus aurata,Sparidae, Linnaeus) larvae

The success of microdiets commonly used in the cultivation of marine fish larvae is limited to serving as partial replacements for live food. This limited success is thought to be associated with a reduced digestive ability due to an incompletely developed digestive system. The enhanced growth obtained from live food has been partially attributed to the digestive enzyme activity of the food organism. The present study was designed to test the effect of an exogenous digestive enzyme incorporated. into a microdiet on the growth of Sparus aurata. Larval gilthead seabream, 20–32 days old, were fed ¹⁴C labelled microdiets containing a commercial pancreatic enzyme at different concentrations (0, 0.1 and 0.05g / 100 g dry diet). Rates of ingestion and assimilation were measured and their relationship to dry weight was determined. Our results show that the success of the microdiet as a food for larval gilthead seabream was limited by the larva's low ingestion rate which only approached its maintenance requirement. In addition, the presence of digestive enzyme in the microdiet enhanced its assimilability by 30%. Larval growth over ten days was 0, 100 and 200% on microdiet free of added enzymes, one with added enzymes and a live food regime, respectively. It is our opinion that successful development of microdiets for Sparus aurata must be based on diets improved both in digestibility and attraction to the larvae. Further studies are now underway to determine the nutritional requirements of gilthead seabream larvae using the experimental method developed in the present study. This research was carried out in partial fulfillment of the requirements for the M.Sc. degree.     

Characterization of digestive enzyme activity during larval development of gilthead seabream (Sparus aurata)

The evolution of the digestive enzyme equipment in seabream from hatching to 30 days old larvae was studied; there was a progressive increase in the activity of protease, amylase and acid and alkaline phosphatase from day 15 onwards. The use of specific inhibitors, and SDS-PAGE provided evidence to suggest that most of the proteases belonged to the serine group. A high -amylase activity was also denoted. Zymograms of larval extracts indicated that exogenous food has more a qualitative than a quantitative role in the secretion of digestive enzymes in this species.     

Reproductive performance of gilthead seabream (Sparus aurata L., 1758) fed two combined levels of carotenoids from paprika oleoresin and essential fatty acids

Despite carotenoids and essential fatty acids seem to play important roles in fish reproduction, no studies have yet been conducted to determine the effect of dietary carotenoids on gilthead seabream broodstock performance and their relation as antioxidants with dietary n‐3 highly unsaturated fatty acid (HUFA) levels. In addition, the high cost of synthetic sources of carotenoids is leading to the search for new natural carotenoid sources such as paprika oleoresin. Four experimental diets containing two combined levels of carotenoids from paprika oleoresin (40 and 60 mg kg^?1) and n‐3 HUFA (25 and 40 g kg^?1) were respectively fed to triplicate groups of gilthead seabream (Sparus aurata) broodfish. Elevation of n‐3 HUFA dietary levels from 25 to 40 g kg^?1 significantly improved gilthead seabream broodstock performance in terms of egg viability, hatching rates and fecundity. Besides, it markedly increased egg contents in HUFAs which play important energetic and structural roles and improve embryo development. Both arachidonic acid and eicosapentaenoic acid (20:5n‐3) egg contents were more readily affected by dietary n‐3 HUFA than docosahexaenoic acid. HUFA levels did not caused any negative effect suggesting an optimized content of antioxidants in broodstock diets. Increase in dietary carotenoids from 40 to 60 mg kg^?1 increased carotenoid contents in eggs and significantly improved egg fertilization rates suggesting an important sperm cell’s protective role by reducing the risk of lipid peroxidation which is detrimental for sperm motility. The increased inclusion of dietary paprika oleoresin enhanced egg carotenoid deposition and improved fish reproductive performance, denoting the high nutritional value of this product as a source of carotenoids for broodstock of this species.     

Preliminary studies on the cryopreservation of gilthead seabream (Sparus aurata) embryos

Vitrification could provide a promising tool for the cryopreservation of fish embryos. In order to achieve successful cryopreservation, several parameters should be taken into account in the design of a vitrification protocol. In the present study some relevant factors were investigated (permeable and non-permeable cryoprotectant toxicity, toxicity of vitrificant solutions, adequate container for embryo loading and temperature of thawing) using two gilthead seabream embryonic development stages (tail bud and tail-bud-free). Permeabilized embryos were incubated in dimethyl sulfoxide (DMSO), methanol (MeOH), ethylene glycol (EG) and 1,2-propanediol (PROH) in concentrations ranging from 0.5 to 6 M for 10 and 30 min and in 5%, 10% and 15% polyvinyl pyrrolidone (PVP), 10%, 15% and 20% sucrose or 0.1%, 1% and 2% X1000® for 2 min. After treatment, embryos were washed and incubated in seawater until hatched. The toxicity of permeable cryoprotectants increased with concentration and exposure time. EG was best tolerated by the embryos. Exposure to non-permeable cryoprotectants did not affect the hatching rate except at F stage. Six vitrificant solutions (DMSO—V1, V2 and V3 and EG—V1, V2 and V3) were tested using a stepwise incorporation protocol. The DMSO-based solutions contained 5 M DMSO + 2 M MeOH + 1 M EG plus 5% PVP, 10% sucrose or 2% X1000® and the EG-based solutions contained 5 M EG + 2 M MeOH + 1 M DMSO plus 5% PVP or 10% sucrose. Before loading the embryos into 0.5 ml straws or 1 ml macrotubes, toxicity tests were effected with these solutions. Our results demonstrated that DMSO-based solutions were better tolerated by seabream embryos than EG-based solutions. After thawing (water bath, 0 or 25 °C), embryos were evaluated by stereoscopic microscopy and the percentage of embryos with intact morphology was registered. The highest percentage of embryos with intact morphology (28%) was observed in samples frozen in macrotubes and thawed at 25 °C. Several malformations associated with ice crystal formation inside the embryos were detected. None of these embryos achieved hatching. Our results suggest that the absence of a proper incorporation of cryoprotectants prior to vitrification is the main problem that must be overcome. This procedure should be optimized in order to avoid ice crystal formation inside embryo compartments.     

Settling velocity and total ammonia nitrogen leaching from commercial feed and faecal pellets of gilthead seabream (Sparus aurata L. 1758) and seabass (Dicentrarchus labrax L. 1758)

The physico-chemical characteristics of particulate wastes of Sparus aurata and Dicentrarchus labrax were investigated. Changes in the dimensions, settling velocity and total ammonia nitrogen (TAN) leached from commercial feed pellets were investigated after soaking. Also, the settling velocity and TAN leached from faecal pellets of these fish were assessed at 15 and 25 °C. The settling velocity of feed pellets was influenced positively by pellet weight and negatively by immersion length as a result of changes in pellet dimensions after soaking. The settling velocity of faecal pellets was determined by pellet weight. The experimental design did not allow identification of any consistent effect of water temperature on settling velocity. Total ammonia nitrogen leaching over time from feed and faecal pellets was successfully explained by means of a first-order kinetic equation. For feed pellets, water temperature significantly affected the speed of the process and the time at which the maximum TAN leached was reached, but did not influence the maximum TAN leached. Leaching was related to feed pellet size, and so the smaller the pellet, the higher the leaching. Total ammonia nitrogen leaching from faecal pellets was greater per unit weight than in feed pellets. However, neither water temperature nor fish species influenced TAN leaching from faeces.     

Enrichment of Artemia nauplii in essential fatty acids with different types of liposomes and their use in the rearing of gilthead sea bream (Sparus aurata) larvae

The present study aimed to optimize the enrichment protocols of Artemia nauplii with highly unsaturated fatty acid liposomes. Three different types of liposomes formulated with krill phospholipid extract were tested, using a commercial emulsion as the control treatment. The incubation time of the nauplii in the enrichment medium was chosen as the first variable studied. Total lipid, lipid class and fatty acid profiles were analyzed in enriched nauplii after 18 and 21 h incubation in the enrichment media. A third sampling point at 24 h, where nauplii had been kept in clean seawater at 28 °C for 3 h, was established to emulate potential changes in the nutritional value of the nauplii after some hours in the larval rearing tank. A second objective of the study was the establishment of a suitable liposome-based diet for the rearing of gilthead sea bream larvae, as a model of marine fish.Differences in the enrichment efficiencies through time were found not only between liposome and emulsion based diets, but also among the different types of liposomes. Whereas nauplii enriched with multilamellar vesicles and unilamellar liposomes prepared by extrusion had maximum enrichment performance at 18 h of incubation, nauplii enriched with unilamellar liposomes prepared by the detergent solubilization method followed the enrichment pattern of the control emulsion, with an optimal incubation period closer to 21 h. Analyses of the diets of the larval nutrition trial seemed to confirm the results obtained in the first experiment, although changes in certain variables occurred among the enrichments carried out during the larval feeding trial. Biochemical characterization of body lipids of the larvae indicated differences in total lipid and essential fatty acid content, although no significant differences were found in growth and survival of the larvae. Thiobarbituric acid reactive substances (TBARS), used to determine the peroxide content of the larvae, indicated a significant higher content of these compounds in larvae fed emulsion enriched nauplii compared to larvae fed liposome enriched prey.     

Pigmentation of gilthead seabream, Sparus aurata (L. 1875), using Chlorella vulgaris (Chlorophyta, Volvocales) microalga

A feeding experiment was conducted over 9 weeks with seven groups of 30 (fish per group) unpigmented gilthead seabream, Sparus aurata (L. 1875) (initial mean weight = 145.2 ± 12.3 g). Three experimental diets were prepared by adding to a basal diet free of carotenoid (final pigment content of around 40 mg per kg feed): (i) a biomass of the carotenogenic Chlorella vulgaris (Chlorophyta, Volvocales); (ii) a synthetic astaxanthin; and (iii) a mixture (1:1) of microalgal biomass and synthetic astaxanthin. At 3‐week intervals, five fish were sampled from each tank for total carotenoids analysis in skin and muscle. The carotenoid pigments (total amount = 0.4%) identified in the carotenogenic alga were lutein (0.3%), β‐carotene (1.2%), canthaxanthin (36.2%), astaxanthin, free and esterified forms (55.0%), and other pigments (7.3%). Carotenoid pigments were significantly deposited in the four skin zones studied during the feeding trial: the forefront between the eyes, the opercule, along the dorsal fin and in the abdominal area. In the muscle, regardless of the astaxanthin source, the amount of carotenoids measured was very low (less than 1 mg kg^?1) and differences not significant. Moreover, no muscle pigmentation was evident, and there was no variation in the amount of carotenoid analysed in skin tissue, through the trial, for each treatment. It was concluded that supplementing the feed with C. vulgaris would be an acceptable practice in aquaculture to improve the market appeal of the gilthead seabream.     

Association of a lordosis-scoliosis-kyphosis deformity in gilthead seabream (Sparus aurata) with family structure

Skeletal deformities constitute a major problem for aquaculture industry by decreasing the final value of the fish. An analysis of skeletal malformations in 11,640 fish was performed considering families and triplicates per family. Thirtynine different skeletal abnormalities were detected, such as lordosis, vertebral fusion, absence of one or both operculum, bent-jaw, etc. Moreover, a new unusual complex spinal column deformity consisting of a consecutive repetition of lordosis, scoliosis and kyphosis (LSK) from the head to the caudal fin was described. This syndrome was statistically associated with the family structure (Z_{2H–family,LSK}=3.49; p<0.05). The incidence of this deformity was 0.2% in the whole population, and 6.5% within the affected family (2H). The environmental and genetic causes are discussed.     

Enrichment of gilthead seabream (Sparus aurata L.) diet with microalgae: effects on the immune system

The present study assessed the effects of three orally administered microalgae (Nannochloropsis gaditana, Tetraselmis chuii and Phaeodactylum tricornutum) on different immune parameters and immune-related gene expression of gilthead seabream (Sparus aurata L.). Fish were fed a control or one of six experimental diets and sampled at 2 and 4?weeks of treatment. At the end of the trial, growth performance and different systemic and local immune activities were measured (natural haemolytic complement activity, IgM levels, phagocytosis and respiratory burst). The expression levels of different immune-associated genes (EF-1α, IgM_H, TCR-β, MHCIα, MHCIIα, CSF-1R and β-defensin) were analysed in head-kidney and gut. Administration of N. gaditana and T. chuii resulted in a significant increase in haemolytic complement activity, phagocytic capacity, as well expression level of β-defensin, as well as MHCIIα and CSF-1R, respectively. The P. tricornutum-supplemented diet provoked immunostimulation, and very little effect on gene expression was observed. These results suggest that dietary microalgae enhance gilthead seabream defence activity, which could be very important in order to consider such microalgae as a possible additive in fish diets.     

Effect of an experimental microparticulate diet on the growth, survival and fatty acid profile of gilthead seabream (Sparus aurata L.) larvae

In recent years, a great deal of interest has emerged in the development of microdiets as an economic alternative to live food, in the larval culture of marine fish species. The ability to grow Sparus aurata larvae on a prototype microparticulate diet was examined. To achieve this objective, four feeding regimes differing in the time when the microdiet was introduced (3, 7 or 12 days) and one based exclusively on an inert diet were tested, during the first 22 days of larval life. Significant differences in larval growth were found between the experimental feeding regimes and their corresponding controls (enriched rotifers during the whole experimental period); the larvae in the co-feeding regimes and with an exclusive microparticulate diet were always significantly smaller than larvae fed on rotifers alone. However, the difference was minimised by introducing the inert diet at a later date. A lower survival was found in larvae with a co-feeding regime, in comparison with the control treatments and the survival was significantly lower in larvae fed exclusively on a microparticulate diet. The fatty acid analysis revealed that the experimental microencapsulated diet and the rotifers enriched with Protein Selco® presented relatively similar fatty acid content. In spite of the slightly higher (n?3)/(n?6) and Docosahexaenoic acid (DHA)/Eicosapentaenoic acid (EPA) ratios and somewhat lower highly unsaturated fatty acid (HUFA) content found in the inert diet, the fatty acid composition of the diets cannot explain the differences found in larval performance. The results revealed that the complete replacement of live prey with the tested microparticulate diet is still not possible in S. aurata larval rearing. Nevertheless, better growth and survival results and a substantial reduction in the daily supply of live food can be achieved with a combination of microdiet and live prey.     

Energy and protein requirements for maintenance and growth in gilthead seabream (Sparus aurata L.)

Factorial determinations of energy and protein requirements in growing Sparus aurata were carried out at 23–24°C. The energy content in the whole fish was dependent on fish weight and ranged from 5 to 11 MJ kg⁻¹ body mass for 1–250 g fish, whereas the protein content remained constant at 179 g kg⁻¹.
During starvation the fish lost 42.5 kJ body weight (BW) (kg)^−0.83 day⁻¹ and 0.42 g protein BW (kg)^−0.70 day⁻¹. The maintenance requirement for energy was calculated to be 55.8 kJ BW (kg)^−0.83 day⁻¹ and for protein 0.86 g BW (kg)^−0.70 day⁻¹. Utilization of digestible energy and digestible crude protein below and at maintenance was determined as 0.72 and 0.51, respectively. Utilization of digestible energy and digestible crude protein for growth above maintenance was determined as 0.46 and 0.28, respectively.
These values allow estimation of requirements for growing Sparus aurata .     

Interference of Listonella anguillarum with potential probiotic microorganisms isolated from farmed gilthead seabream (Sparus aurata, L.)

Four isolates obtained from gilthead seabream have been tested for their adhesion to the skin, gill and intestinal mucus of gilthead seabream, and for their ability to interfere with Listonella anguillarum, an important pathogen of farmed gilthead seabream. The ability to adhere to mucus was higher than 7% for all isolates. Three isolates showed an antagonistic effect against some of the pathogenic strains tested. They were assayed to interfere with the attachment of L. anguillarum to the mucus of gilthead seabream. Only two isolates significantly reduced the adhesion of L. anguillarum to all of the mucus assayed under exclusion, competition and displacement conditions. According to the criteria applied, the isolate Pdp11 was selected and its in vivo probiotic potential was assessed by oral administration followed by challenge with the pathogen L. anguillarum. For the feeding trial, a group of 50 gilthead seabreams received a commercial diet supplemented with lyophilized 10⁸ CFU g^?1 of this isolate for 15 days. An other group of similar characteristics received the non‐supplemented commercial diet. After the challenge, the mortality of the fish receiving the diet supplemented with the potential probiotic was significantly lower than that observed in the groups of fish receiving the non‐supplemented commercial diet.     

Ovarian contractility in reared gilthead seabream (Sparus aurata L.) in different phases of the reproductive cycle

Spontaneous ovarian tunica albuginea contractility was evaluated in gilthead seabream (Sparus aurata L.) at different phases of the reproductive cycle. Fourteen adult females were sampled from February to November 2012 in a commercial fish farm, and ovaries were removed and processed for histological and contractility analyses. Fish reproductive stages were evaluated on haematoxylin–eosin-stained ovary sections or by simple macroscopic observation of hydrated oocytes in spawning individuals. Tunica albuginea spontaneous contractility was recorded by using ovary wall strips mounted in an organ bath containing modified Ringer’s solution. Ovary macro- and microscopic analyses allowed the identification of three different reproductive conditions: vitellogenesis, spawning and regressing. The gilthead seabream tunica albuginea was capable to contract spontaneously, and significant differences were found in mean contraction amplitude among the three reproductive states, with the highest value recorded in individuals in regressing condition and the lowest in individuals at spawning stage. No differences in mean contractility frequency among the three different groups were found. Possible involvement of spontaneous contractility in facilitating developing follicle advancement towards the ovarian lumen within the ovary and in supporting recovery of regressing ovaries may be hypothesized. The low contractility observed during the final oocyte maturation and spawning phases does not seem to support a role of tunica albuginea during ovulation, which could conversely involve theca cell contraction. Alternatively, possible single instantaneous contractions of tunica albuginea muscle fibres, not detected in the present study, could occur during ovulation in response to neuro-hormonal stimulations; a role of abdominal wall musculature in ovary “squeezing” and consequent release of ovulated eggs cannot be excluded.     

Mono- and duoculture of juvenile sharpsnout seabream Diplodus puntazzo (Cetti) and gilthead seabream Sparus aurata L. in a recirculated water system

Although the growth of intensively reared sharpsnout seabream Diplodus puntazzo (DP) has been suggested to improve in duoculture with gilthead seabream Sparus aurata (SA), the stocking ratio for most effective duoculture rearing has not been investigated. For this purpose, juvenile D. puntazzo (1.68±0.011 g) and S. aurata (1.43±0.007 g) were reared in duplicated groups of 165 specimens each, in two monoculture (100% DP, 100% SA) and four duoculture combinations (80% DP–20% SA, 60% DP–40% SA, 40% DP–60% SA, 20% DP–80% SA) for 96 days under recirculated water system conditions. Both species exhibited the highest growth when stocked at 20% and the poorest growth when stocked at 80%. For each species, no major differences for coefficient of weight variation and carcass proximate composition were detected. As the percentage of stocked S. aurata increased, D. puntazzo aggressiveness and weight variability of whole population were reduced, while total biomass, food consumption and food conversion ratio were increased. The results obtained are discussed in relation to fish behaviour and social interactions. From the fish farming point of view, it is indicated that under the present experimental conditions, juvenile S. aurata should be reared in monoculture and D. puntazzo in duoculture 40% DP–60% SA.