Incorporation and metabolism of14C-labelled polyunsaturated fatty acids in wild-caught juveniles of golden grey mullet,Liza aurata,in vivo

The incorporation, and the capacity for desaturation and elongation in vivo, of intraperitoneally-injected,¹⁴C-labelled n-3 and n-6 C₁₈ and C₂₀ polyunsaturated fatty acids (PUFA) were investigated in juvenile golden grey mullet,Liza aurata. The results indicate that juvenile mullet have only limited ability to convert C₁₈ polyunsaturated fatty acids to C₂₀ and C₂₂ highly unsaturated fatty acids (HUFA)in vivo. This suggests that juvenile golden grey mullet require the provision of preformed C_20/22 HUFA, such as eicosapentaenoic and docosahexaenoic acids, in the diet. The impairment in the desaturase/elongase pathway was similar to that found in turbot,Scophthalmus maximus, and gilthead sea bream,Sparus aurata, being primarily at the level of Δ5-desaturase. The data from the largely herbivorous golden grey mullet juveniles are consistent with the hypothesis that marine fish in general, irrespective of dietary habits, have limited capacity for the desaturation and elongation of C₁₈ PUFA. The defect in Δ5-desaturase activity combined with the consistent finding that arachidonic acid is selectively incorporated and retained in membrane phosphatidylinositol suggests that, like turbot and gilthead sea bream, golden grey mullet may also have a requirement for preformed arachidonic acid in the diet.     

Effects of dietary polyunsaturated fatty acid/vitamin E (PUFA/tocopherol ratio on antioxidant defence mechanisms of juvenile gilthead sea bream (Sparus aurata L., Osteichthyes, Sparidae)

Lipid peroxidation, specifically polyunsaturated fatty acid (PUFA) oxidation is highly deleterious, resulting in damage to cellular biomembranes, and may be a principal cause of several diseases in fish including jaundice and nutritional muscular dystrophy. Tissue lipid PUFA content and composition are critical factors in lipid peroxidation, as is the level of endogenous antioxidant molecules such as vitamin E. The primary objective of the present study was the characterization of antioxidant systems in a cultured juvenile marine fish, gilthead sea bream (Sparus aurata) with the underlying aim to understand how to avoid oxidation problems that may cause pathologies and disease and so to enhance growth and quality of early ongrowing stages. Juvenile sea bream were fed diets having either high or low levels of fish oil and supplemented or basal levels of vitamin E with PUFA/vitamin E ratios ranging from 117±12 in the diet with low PUFA supplemented with vitamin E to 745±48 in the diet with high PUFA with no additional vitamin E. None of the diets had serious deleterious effects on growth or survival of the fish, but the different dietary regimes were effective in significantly altering the PUFA/vitamin E ratios in the fish livers with values ranging from 5.7±0.4 in fish fed the diet with low PUFA supplemented with vitamin E to 91.1±13.2 in fish fed the diet with high PUFA with no additional vitamin E. This had effects on the peroxidation status of the fish as indicated by the significantly altered levels of in vivo lipid peroxidation products measured in liver, with fish fed the diet rich in PUFA and low in vitamin E showing significantly higher values of thiobarbituric acid reactive substances (TBARS) and isoprostanes. The isoprostane levels generally followed the same pattern as the TBARS levels supporting its value as an indicator of in vivo oxidative stress in fish, as it is in mammals. However, few significant effects on antioxidant enzyme activities were observed suggesting that more severe conditions may be required to affect these activities such as increasing the PUFA/vitamin E ratio or by increasing peroxidative stress through the feeding of oxidized oils.     

The desaturation and elongation of 14C-labelled polyunsaturated fatty acids by pike (Esox lucius L.) in vivo

To examine the ability of pike (Esox lucius L.) to modify exogenous PUFA by desaturation and elongation, ¹⁴C-labelled 18:2(n-6), 18:3(n-3), 20:4(n-6) and 20:5(n-3) were injected intraperitoneally and the distribution of radioactivity in tissue lipid classes and liver PUFA measured. In all tissues examined, radioactivity from all ¹⁴C-PUFA was recovered in many classes of acyl lipids and the level of recovery generally reflected the relative abundance of the lipid classes. Triacylglycerols, CGP and EGP usually contained high levels of all incorporated ¹⁴C-PUFA. PI contained higher levels of radioactivity from ¹⁴C-20:4(n-6) than from other injected substrates. In liver lipid, the 6 desaturation products of ¹⁴C-18:2(n-6) and ¹⁴C-18:3(n-3) contained no measurable radioactivity although the elongation products of the 6 desaturation products were labelled, as were the direct elongation products of these injected substrates. No radioactivity from ¹⁴C-18:2(n-6) or ¹⁴C-18:3(n-3) was detected in C₂₀ or C₂₂ products of 5 and 4 desaturation. Almost all radioactivity from injected ¹⁴C-20:4(n-6) was recovered in this PUFA. Of the total radioactivity from ¹⁴C-20:5(n-3) incorporated into liver lipid, 7% was present as 24:5 and 16.4% was recovered in hexaenoic fatty acids. In liver, 24:5(n-3) and 24:6(n-3) each accounted for 1% of the mass of total fatty acids and were located almost exclusively in triacylglycerols. The presence of radioactivity in these C₂₄ PUFA suggests that in pike the synthesis of 22:6(n-3) from 20:5(n-3) may proceed without 4 desaturase via the pathway which involves chain shortening of 24:6(n-3). It is concluded that under the circumstances employed in this study pike, do not exhibit 5 desaturase activity and are unable to synthesize 20:4(n-6) and 20:5(n-3) from 18:2(n-6) and 18:3(n-3), respectively. This suggests that pike may require 20:4(n-6) and 20:5(n-3) preformed in the diet.Abbreviations CGP choline glycerophospholipids - CL cardiolipin - EGP ethanolamine glycerophospholipids - PG phosphatidylglycerol - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acids - SM sphingomyelin - TLC thin-layer chromatography     

Comparison of the swimming performance of farmed and wild gilthead sea bream, Sparus aurata

Farmed gilthead sea bream, Sparus aurata, frequently escape from the sea cages and interact with wild populations. The impact of these interactions on the wild populations will depend, in part, on differences in performance of the bream. This study compared the swimming performance of the wild and farmed fish in a current channel. The absolute critical swimming speed (U_crit) increased with increasing size while the relative U_crit decreased. Even at the same length there were noticeable performance differences between the individuals. The wild sea bream have significantly higher (P<0.05) absolute U_crit performance (0.86±0.01 m s⁻¹) than the farmed fish (0.79±0.01 m s⁻¹) and significantly higher (P<0.05) relative U_crit performance (4.52±0.05 BL s⁻¹) than the farmed fish (4.21±0.05 BL s⁻¹). The present study suggests that cultured sea bream may not have the ability to compete with wild sea bream in native seawaters.     

Effect of broodstock diets on lipid classes and their fatty acid composition in eggs of gilthead sea bream (Sparus aurata L.)

Total lipid content, total lipid fatty acids, lipid classes and their fatty acid compositions were studied in eggs of gilthead sea bream from two different broodstocks. The two broodstock diets used had the same total lipid content, but differed with respect to their fatty acid levels. The lipid class composition of eggs from different origins was not significantly different. The predominant polar lipids were phosphatidylcholine, phosphatidylethanolamine and sphingomyelin, and the main neutral lipid classes were cholesterol esters and triacylglycerols. The fatty acid composition of total lipids and main lipid classes in the two groups of eggs showed marked differences, reflecting the influence of fatty acid levels in the broodstock diets.     

Effect of broodstock diets on total lipids and fatty acid composition of larvae of gilthead sea bream (Sparus aurata L.) during yolksac stage

Total lipid content and total lipid fatty acid compositions were studied in larvae at the yolksac stage of gilthead sea bream from two different broodstocks. The two broodstock diets had the same total lipid content but were different with respect to their fatty acid levels. There were differences in the fatty acid composition of total lipids in the two groups of larvae, reflecting the influence of fatty acid levels in the diets fed to the broodstock. The main fatty acids in both larvae lots during yolksac stage were 16:0, 18:1n-9 and 22:6n-3. The desaturation index Δx = [P + Σ (n − x) − Σ (n − x) diet]/P allowed the determination of the relative importance of the Δ9, Δ6, Δ5 and Δ4 desaturations during the egg and larval yolksac stages. The levels of total lipid fatty acids in eggs and larvae during yolksac period were almost constant and very low desaturation and elongation activities were apparent. A competitive inhibition was observed between the fatty acids of the n-6 series with respect to the ones of the n-3 series in those eggs and larvae from broodstock whose diet contained a high proportion of linoleic acid and its derivatives.     

Evaluation of immunological parameters in farmed gilthead sea bream, Sparus aurata L., before and during outbreaks of 'winter syndrome'

In recent years, 'winter syndrome' has caused severe economic losses in many marine farms in southern Europe. This study compared the activity of the immune system in healthy, asymptomatic and diseased gilthead sea bream. Serum protein and immunoglobulin content were evaluated as well as serum complement activity from these groups of fish. Differential leucocyte counts were also determined. Immunological assays were performed to determine intra- and extracellular 'respiratory burst' activity as non-specific immune response parameters. Diseased fish showed a significant reduction in haemolytic activity (90%), and in serum proteins and immunoglobulins in comparison with the other groups. A significant increase in the lymphocyte percentage (50%) and a decrease in the granulocyte percentage (over 70%) was found in asymptomatic fish. Respiratory burst activity was reduced in both the clinical and preclinical stages of the disease, compared with the controls. These observations confirm a severe immunodeficiency in diseased fish but also the presence of a cellular immune dysfunction in fish without clinical signs, before the onset of the disease.     

Recent advances in European sea bass and gilthead sea bream nutrition

European sea bass (Dicentrarchus labrax) and gilthead sea bream(Sparus aurata) are amongst the most important finfish speciescultured in the Mediterranean region. Production of these species isnowadays a well-controlled process, but knowledge of their nutritionalrequirements is still very limited. Nevertheless, a considerable amountof data has been accumulated in recent years, and the purpose of thispaper is to review the recent advances on the nutritional requirementsof sea bass and sea bream. The optimum protein to energy ratio of thediets of sea bass and sea bream seem to be higher than for salmonids,and there is some evidence that high dietary lipid levels have nobeneficial effects on fish performances. Although the essential aminoacid requirements were estimated by the ideal protein method, data basedon the dose-response method is only available for a few amino acids.Essential fatty acid requirements were estimated for sea bream juvenilesbut data is lacking for sea bass. Vitamin and mineral requirements ofthese species are practically unknown. Although the importance ofbroodstock nutrition on gonadal development, spawning and egg quality isrecognized, few studies were done to elucidate these aspects. The recentdevelopment of microparticulate diets for larvae will contribute to theaccurate evaluation of their nutritional requirements.     

Glomerulonephritis and immunosuppression associated with dietary essential fatty acid deficiency in gilthead sea bream, Sparus aurata L., juveniles

An experiment was conducted to determine the effect of an essential fatty acid (EFA) deficient diet on growth, immune status and renal morphology of juvenile gilthead sea bream, using two diets: a control diet containing 2% (DW) of n-3 high unsaturated fatty acid (n-3 HUFA) and a diet formulated to be deficient in EFA for this species and containing 0.5% DW of n-3 HUFA (diet NFA). After 9 weeks of feeding the EFA-deficient diet fish showed a reduction in growth compared with the control group (107.48 +/- 9.14 and 123.14 +/-11.87 g final weight respectively). Fish fed the NFA diet showed a reduction in the erythrocyte volume together with increased erythrocyte fragility, haemoglobin content and red blood cell count in comparison with fish fed the control diet. EFA deficiency also reduced cellular immunity in terms of neutrophil activity and the number of circulating lymphocytes. The serum alternative complement pathway was markedly reduced in fish fed the EFA deficient diet. Fish fed the control diet showed glomeruli with a well-defined Bowman's space and normal renal tubes. However, up to 88% of fish fed the EFA-deficient diet showed alterations in renal morphology affecting at least 50% of the glomeruli, which showed extreme dilation of capillaries and occlusion of Bowsman's capsule. Mesangial proliferation and diffuse thickening of the capillary walls, as well as renal tube degeneration, were also observed.     

The effect of food competition and relative size of group members on growth of juvenile gilthead sea bream, Sparus aurata

The effects of food competition and relative size of group members on growth of sea bream, Sparus aurata were tested in groups composed of four fish. Each group consisted of a medium sized test fish (approximately 250 mg) and three fish larger or smaller than the test fish. In the first experiment food competition was addressed directly by contrasting its presence with its absence. Neither food competition nor relative size of school members had a significant effect on growth. However, there was a highly significant interaction between these two main effects. Under conditions of food competition the medium sized test fish had a three times higher specific growth rate when surrounded by small vs large fish. Under conditions of no food competition when the test fish were separated during feeding a similar but reverse pattern occurred due to differences in food conversion efficiency. In the second experiment, the effects of food quantity and relative size of group members were tested. Both main effects and their interaction were highly significant. Under conditions of low food competition (i.e., surplus food supply) fish grew again better when surrounded by larger fish as compared to their growth rate when surrounded by smaller ones. Size matching via a differential growth rate in a schooling species such as sea bream was suggested to increase the confusion effect by reducing the ability of a predator to focus on an odd sized fish. The implications of direct competition for food for the efficient culture of this species are discussed.     

Effects of dietary mannan oligosaccharides in early weaning diets on growth,survival, fatty acid composition and gut morphology of gilthead sea bream (Sparus aurata,L.) larvae

Early weaning of marine fish larvae with dry diets delays gut maturation and reduces growth rates. In juvenile and adult forms of several marine fish species, inclusion of dietary mannan oligosaccharides (MOS) improves gut integrity and functionality, but the effects of MOS inclusion in gilthead sea bream (Sparus aurata, L.) larval diets have not been addressed yet. Thus, this study assesses the effects of dietary MOS inclusion on survival, growth performance, gut morphology, feed acceptance and quality of gilthead sea bream larvae. For that purpose, 16 days post‐hatched gilthead sea bream larvae were fed four graded levels of MOS (Biomos^®, Alltech, Nicholasville, KY, USA) in weaning diets as follows: 0 g kg^?1 MOS, 0.5 g kg^?1 MOS, 1.5 g kg^?1 MOS and 2 g kg^?1MOS. Dietary MOS did not affect feed acceptance in gilthead sea bream larvae (P > 0.05). MOS supplementation was correlated in a dose‐dependent way with higher larval survival (P = 0.026). After 15 days of feeding, dietary MOS increased whole larvae (P < 0.01) arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid. Gilthead sea bream larvae fed 2 g kg^?1 MOS presented higher gut occupation with goblet cells after feeding compared with larvae fed the other dietary treatments. Overall, the results suggest that inclusion of MOS in early weaning diets for gilthead sea bream improves essential fatty acid utilization and may promote growth and final survival.     

Effects of protein level of the diet on digestibility and growth of gilthead sea bream, Sparus auratus L.

A feeding trial with a duration of 12 weeks was conducted to determine the effects of various protein levels, in relation to diet digestibility and growth of the gilthead sea bream, Sparus aurata L. Four experimental fish meal/wheat meal based diets (A,B,C,D) containing 400, 450, 500 and 550 g kg^?1 protein, respectively, were tested. The increase of the fishmeal content of the diet led to an increment of dry matter digestibility. Apparent digestibility coefficients of protein and lipids were always very high (being near or even over 90%). Energy digestibility coefficients increased from diet A to diet D, which corresponds to a decrease in the wheat meal content of the diet. Voluntary feed intake increased with the decrease of protein content of the diet (from diet D to diet A). In the other sense, feed/gain ratio decreased regularly as protein percentage increased (from diet A to diet D). The most favourable feed/gain ratio, 1.07, was noted for the group receiving 55% protein (diet D). Fish on the lowest protein diet (Diet A) showed the highest protein efficiency ratio (PER) and the highest percentage retention of the digestible protein intake. Other than slight positive differences between fish fed diets with 500 and 550 g kg^?1 protein, no significant differences were observed for growth when dietary protein exceeded 450 g kg^?1. Beyond this level, no significant difference in final average individual weight was observed. Although it is generally considered that the dietary protein requirement for gilthead sea bream is 400 g kg^?1, our experiment demonstrates that to obtain high growth rates (>2.3% per day), a minimum of 450 g kg^?1 protein in the diet is necessary. However, the most favourable values for growth rates and feed/gain ratio are obtained with 550 g kg^?1 protein in diet, considering that no significant differences were observed for PER among diets B, C and D.     

Microsatellite markers reveal population structure in gilthead sea bream Sparus auratus from the Atlantic Ocean and Mediterranean Sea

ABSTRACT:   This study was carried out to investigate the genetic variability and the population genetics of Sparus auratus. In fact, despite its importance in Mediterranean fisheries and aquaculture, very little it is known concerning its population structure. Samples of wild gilthead sea bream were collected in seven different localities along the Mediterranean and Atlantic coasts and were genetically characterized by means of microsatellite markers. Genotyping at four microsatellite loci revealed high polymorphism (7–38 alleles/locus) and expected heterozygosities, which ranged from 0.80 to 0.85. A slight but significant population structure was found ( F _ST = 0.010). In fact, at least three populations of gilthead sea bream within the Western Mediterranean Sea were identified (Sardinian Sea, Sardinian Channel and Central Tyrrhenian Sea), which are also genetically differentiated from those of the Atlantic Ocean and the Adriatic Sea.     

Effect of dietary energy content on gilthead sea bream (Sparus aurata) feeding behaviour and nutritional use of the diet

We have studied how the diet energy level affects gilthead sea bream (Sparus aurata) daily pattern of demand‐feeding activity and nutritional use of the diet under summer‐ and winter‐like conditions. To that end, animals were kept in a closed circuit under controlled temperature and photoperiod, and fed one of two commercial diets with either high (H: 238 g kg^?1 fat) or low (L: 172 g kg^?1 fat) energy content. In summer conditions (26 °C; 12 : 12 L : D), both diets yielded similar growth rate (0.7 ± 0.0 and 0.7 ± 0.1) and thermal unit growth coefficient values (0.6 ± 0.0 and 0.6 ± 0.0), and although the daily rate of delivered feed was somewhat higher for fish under the high‐energy diet (16 ± 2 g kg^?1 of fish) the difference was not significant. In winter conditions (17 °C; 9 : 15 L : D), on the other hand, no differences were found for any of the parameters evaluated. Thus, the higher lipid content of the diet does not appear to have any protein sparing effect. The ‘summer’ demand‐feeding pattern displayed three daily peaks, which were quite apparent for fish under the low‐energy diet, but rather smoothed for those that ate the energy‐rich diet. This profile was replaced in winter conditions by a single peak around noon, and the animals also displayed a lower overall number of demands.     

The in vivo incorporation and metabolism of [1-14C] linolenate (18:3n-3) in liver, brain and eyes of juveniles of rainbow trout Oncorhynchus mykiss L and gilthead sea bream Sparus aurata L.

Accumulation of docosahexaenoic acid (DHA; 22:6n-3) in brain and eyes during development has been demonstrated in fish but it is not clear whether liver or neural tissues themselves are of greater importance in the biosynthesis of DHA from dietary 18:3n-3. In the present study, we investigated the in vivo metabolism of intraperitoneally injected [1-¹⁴C]18:3n-3 in liver, brains and eyes of young juvenile fish. Metabolism was followed over a 48h time-course in order to obtain dynamic information that could aid the elucidation of the roles of the different tissues in the biosynthesis and provision of DHA from dietary 18:3n-3. The study was performed in both a freshwater fish, rainbow trout Oncorhynchus mykiss L and a marine fish, gilthead sea bream Sparus aurata L to determine the effect that low or limiting5-desaturase activity may have in this process. As expected, the results showed that although the sea bream incorporated more 18:3n-3 into its lipids, metabolism of the incorporated fatty acid by de saturation and elongation was generally greater in the trout. In liver, the percentages of radioactivity recovered in tetraene and pentaene products were greater in trout than in sea bream although there was no difference in hexaenes. In contrast, the re covery of radioactivity in DHA was significantly greater in brain in trout compared to sea bream. In both species, the percentage of radioactivity recovered in desaturated/elongated products was much lower in liver than in brains and eyes, but that percentage increased over the 48h time-course. In trout though, the highest percentages of desaturated products in brain and eye were observed after 12 and 24h, respectively. However in sea bream the highest percentages of desaturated products in the neural tissues were observed after 24-48h. Radioactivity was recovered in 24:5n-3 and 24:6n-3, intermediates in the 4-independent ("Sprecher shunt") pathway for the synthesis of DHA, in both species, especially in the brain and eyes. Overall, although the results cannot eliminate a role for liver in the biosynthesis and provision of DHA for developing neural tissues in fish, they suggest that DHA can be synthesised in fish brain and eye in vivo.     

Effects of chronic confinement on physiological responses of juvenile gilthead sea bream, Sparus aurata L., to acute handling

Understanding how gilthead sea bream, Sparus aurata L., an important Mediterranean Sea species for aquaculture, respond physiologically to stressors commonly encountered in intensive rearing is important for effective production, as managing for stress is a major factor in maintaining healthy fish stocks. Our objective was to determine whether holding juvenile gilthead sea bream at a high density (HD), as a chronic stressor, would affect their physiological responses to a subsequent acute handling stressor. After acclimation at a low density (LD) of 6 kg m^?3 in 200‐L circular tanks containing 33–36 g L^?1 recirculating seawater at 19°C under a normal photoperiod, juvenile 37‐g gilthead sea bream were confined for 14 days at a HD of 26 kg m^?3 and then subjected to 30‐s aerial emersion in a dipnet. Plasma levels of cortisol, glucose, lactate, osmolality and chloride were determined in fish held in separate lots during LD (control) and HD confinement at 0, 1, 2, 7 and 14 days, and then after handling at 0, 1, 2, 4 and 8 h. Although plasma cortisol levels were similar in LD and HD fish groups after 14 d of confinement (15 and 23 ng mL^?1, respectively), the cortisol response in fish from the HD treatment at 1 and 2 h following acute handling (70 and 37 ng mL^?1, respectively) was only about half of that measured in the control group (139 and 102 ng mL^?1); plasma cortisol was similar in both groups by 4 and 8 h. In contrast, plasma glucose elevations in response to handling were higher at 4 and 8 h in the HD‐held fish (94 and 72 mg dL^?1, respectively) than in those from the LD treatment (59 and 51 mg dL^?1); glucose responses were similar in both groups at 1 and 2 h after handling and throughout confinement. Plasma lactate levels were higher in LD fish than in the HD group at the beginning of the experiment but were similar after 14 d confinement and responses to handling were similar (e.g. 33 and 35 mg dL^?1 at 1 h). Plasma osmolality showed increases during the first 2 h after acute handling but no differences were evident between the two density treatments at any time during confinement or posthandling. Plasma chloride levels did not change throughout the experiment. The reduced plasma cortisol response to acute handling likely resulted from negative feedback of mildly but chronically elevated circulating cortisol caused by the confinement stressor on the hypothalamic–pituitary–interrenal axis. While other post‐handling physiological changes also showed differences between treatment groups, the suppressed cortisol response in the HD‐held fish suggests a reduction in the gilthead sea bream's normal capacity to respond to an acute stressor.     

Starvation before slaughtering as a tool to keep freshness attributes in gilthead sea bream (Sparus aurata)

Instrumental quality parameters related to freshness were evaluated for gilthead sea bream (Sparus aurata) after different storage periods on ice (2, 4, 7 and 11 days of storage) for one, four and eight days of pre-slaughter starvation. The study was carried out on 360 gilthead sea bream from five different groups reared on a commercial fish farm. Starvation period had a significant effect on fish texture: the force necessary to compress 4 mm of flesh after two days ice storage in 1 day starved fish (16.50 ± 0.59 Newtons) was less than that for four day (18.98 ± 0.63 Newtons) and eight day starved fish (20.43 ± 1.12 Newtons). Significant differences in flesh puncture and flesh compression were not detected, although these values did increase with increasing starvation period. A significant increase in pH and myofibrilar protein solubility was also detected for fish starved for one day.     

Biochemical differences in lateral muscle of wild and farmed gilthead sea bream (series Sparus aurata L.)

Red and white muscle from specimens of wild and farmed gilthead sea bream (Sparus aurata) were analyzed for histochemical ATPase activity, total protein content, fatty acids, trace element concentrations and myosin isoforms. The fibre type composition of muscle samples was confirmed histochemically by the ATPase reaction, which did not show any differences between the two groups of animals. Myosin ATPase activities, myosin and protein yields were significantly higher in white muscle than in the red muscle and for the red muscle the latter two parameters were higher in wild fish. Fatty acid profiles revealed differences between the two groups of animals, probably because of the fatty acid composition of the diets. Zinc, copper and iron concentrations were higher in red muscle than in white muscle; muscles from wild fish were significantly richer in trace elements. No separation of fast and slow heavy chains of myosin could be obtained on SDS-gel electrophoresis, but two dimensional electrophoresis revealed the presence of three light chains in white muscle (LC1F, LC2F, LC3F), and two main types in red muscle (LC1S, LC2S). Small, variable percentages of LC3F were found in the red muscle samples, especially in the wild fish. It is concluded that the different environmental conditions, experienced by wild and farmed fish, have significantly influenced the biochemical composition of their lateral muscle.     

Effect of different feeding strategies on gilthead sea bream (Sparus aurata) demand-feeding behaviour and nutritional utilization of the diet

We have studied the effects of time‐restricted food access and ration restriction on gilthead sea bream demand‐feeding behaviour and nutritional use of the diet (Sparus aurata), and also compared the nutritional efficiency of three different feeding systems: manual, automatic and modulated‐automatic. In the first trial, fish were allowed to feed from self‐feeders under three different conditions: ad libitum, ration restriction, and time‐restricted food access, and their demand‐feeding pattern, diet utilization and body composition were analysed. In the second trial, animals were fed by hand or using an automatic system, either fixed or modulated, and diet utilization and body composition were analysed as before. Restricting the amount of food modifies gilthead seabream self‐feeding behaviour, with fish increasing the number of demands provided these are rewarded with food. However, demand‐feeding activity does not increase if rewards are restricted to a certain time. Feeding gilthead sea bream by hand versus automatically, and distributing the daily food ration in two or three equal or unequal‐size daily meals, have no effect on the animals’ growth, nutritional use of the diet or body composition.