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.     

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.     

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.     

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.     

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 high-level fish meal replacement by plant proteins in gilthead sea bream (Sparus aurata) on growth and body/fillet quality traits

Juvenile gilthead sea bream (initial body weight ca. 100 g) were reared in an indoor flow through marine water system for 1 year. Fish were fed two isoenergetic [19.2 kJ g⁻¹ dry matter (DM)] and isoproteic (426 g kg⁻¹ DM) diets either based on fish meal (diet FM) or on a mixture of plant protein sources (diet PP), replacing 75% of fish meal protein. The growth trial was conducted in duplicate, two tanks for each dietary treatment. Growth performance and feed utilization were registered. Fillet quality parameters were evaluated and sensory analyses on cooked fillet were performed. Both groups had similar weight gain and specific growth rates. Feed intake was higher in sea bream fed diet FM (0.48 versus 0.44), while feed efficiency and protein efficiency ratio were significantly higher in sea bream fed PP (0.83 versus 0.77 and 2.0 versus 1.76, respectively). Sea bream fed diet FM had a lower hepatosomatic index (0.80 versus 0.87%), and a higher fillet yield (45.9 versus 44.9%). The fillet from sea bream fed diet FM had higher moisture (696 versus 682 g kg⁻¹), lower lipid levels (91 versus 100 g kg⁻¹) with higher levels of n‐3 polyunsaturated fatty acids (PUFA) and monounsaturated fatty acids (MUFA), while the PP fed sea bream presented a higher level of PUFA n‐6. There were minor differences in muscle free amino acid levels between the two diet groups. As regards sensory evaluation of cooked fillet, the judges were unable to discriminate the two dietary groups of fish. Summarizing, the results demonstrate the possibility to use diets containing high levels (750 g kg⁻¹) of plant ingredients in gilthead sea bream without affecting growth performance and with minor effects on quality traits of commercial size sea bream.     

金头鲷的形态学及框架结构的研究

从初孵仔鱼培育到17月龄鱼种中,检测50尾鱼(鱼龄17个月)的体重(g)、体长(cm)、全长(cm)、尾叉长(cm)、体高(cm)、体宽(cm)、头长(cm)、吻长(cm)、眼后头长(cm)、眼径(cm)、尾柄长(cm)和尾柄高(cm)等的平均值及标准差分别为241·11±49·56、18·72±1·30、24·15±1·39、22·02±1·23、8·20±0·63、3·18±0·31、5·56±0·36、1·98±0·28、2·47±0·21、1·52±0·09、2·56±0·45、1·96±0·17;体长/体高、体长/头长、体长/尾柄长、体长/眼径、头长/眼径、头长/尾柄长、头长/尾柄高、尾柄长/尾柄高、头长/眼后头长和体高/体厚等的平均值分别为2·29±0·10、3·33±0·18、7·49±1·25、12·37±1·12、3·72±0·30、2·26±0·43、2·88±0·25、1·31±0·21、2·29±0·16和2·59±0·22。其可数性状:背鳍、胸鳍、腹鳍、臀鳍等的鳍式为D.Ⅺ-12~14、P1.5 (9~10)、P2.Ⅰ-1 4、A.Ⅲ-11;侧线鳞为56~69枚、侧线上鳞为6~8枚;脊椎骨数为9~10(胸骨) 13~14(尾骨)根;上颌齿式为:1~5 7~10、2~13、4~9、0~9/0~7、3~10、3~15、4~11 1~4;下颌齿式为:1~2 6~15、4~10、0~8/0~7、4~9、6~12 1~2。通过观察,运用传统测量法对其外观的可数、可量性状进行测量与分析,并结合Humphsies,Bookstein和Bookstein等提出的框架法(Truss)思路来构建金头鲷的框架结构图,几何地描述金头鲷的体形特征,可为研究金头鲷的种质提供科学依据。     

Pharmacokinetics of oxolinic acid in gilthead sea bream, Sparus aurata L.

This is the first study on the pharmacokinetic parameters of oxolinic acid (OA) in gilthead sea bream, Sparus aurata L. The kinetic profile of OA was studied after a single intravascular injection (20 mg kg⁻¹) in 100 g fish at 20 °C. The distribution half-life ( t _1/2α) and the elimination half-life ( t _1/2β) of the drug were found to be short (0.51 and 12.60 h, respectively). The drug penetration from the plasma to the tissues was adequate as the apparent volume of distribution of the drug at steady-state ( V _d(ss)) was found to be 2.11 L kg⁻¹. The mean residence time ( MRT ) of OA was short (14.25 h) and the total clearance rate ( Cl _T) of the drug was low (0.15 L kg⁻¹ h⁻¹). The bioavailability ( F %) of OA following oral administration (30 mg kg⁻¹) was also low (14%). Maximum values were observed for muscle at 0.5 h after injection, with levels declining as with subsequent sampling. At the first two time points (0.5 and 1 h) plasma levels of OA were higher than muscle, however, the reverse was evident for subsequent samples. Following oral administration, highest muscle levels were found at 16 h and, with the exception of the 24-h sampling, muscle OA concentrations were higher than plasma at all time points. The fast elimination of OA suggests short withdrawal times with reference to human consumption of treated fish.     

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.     

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.     

Enhancement of gilthead seabream (Sparus aurata) larval growth by dietary vitamin E in relation to two different levels of essential fatty acids

The objective of this study was to determine the effect of dietary vitamin E on gilthead seabream (Sparus aurata) growth and survival, at two different highly unsaturated fatty acids (HUFAs) levels. Eighteen days old gilthead seabream larvae were fed four formulated experimental diets combining two different dietary levels of HUFAs (M: medium 2.5 + 1.5, DHA + EPA, H: high 5 + 2.5 DHA + EPA g per 100 g) with two different levels of vitamin E (M: medium 540 mg kg^?1, H: high 2900 mg kg^?1): MM, MH, HM, HH (HUFA/vitamin E). After 2‐week feeding trial, the average survival rate was 52.6% and there were no significant differences found among treatments. Increase in vitamin E up to high level markedly improved larval growth, particularly when dietary HUFA levels were lower, suggesting a higher protection value when these fatty acids are more limiting. At medium dietary HUFA levels, increase in vitamin E from medium to high level enhanced larval growth performance in terms of total length. Moreover, increase in vit E enhanced HUFAs content in the larval polar lipids denoting the anti‐oxidative effect of vitamin E.     

金头鲷全人工育苗技术研究

利用金头鲷亲鱼202尾,获受精卵39900万粒,选取产卵盛期受精卵583万粒,孵化初孵仔鱼462万尾,平均孵化率78．2％,培育出3—4cm的幼鱼34．2万尾,平均成活率7．4％,最高达18．4％。     

Apparent digestibility coefficients of feed ingredients and their predictability in compound diets for gilthead seabream, Sparus aurata L.

Apparent digestibility of crude protein, amino acids, lipid, carbohydrate and energy was measured for a range of feed ingredients fed to gilthead seabream, Sparus aurata L. — fish meal, poultry meal, meat meal, blood meal, squid meal, extracted soyabean and wheat flour. Chromic oxide was used as a non-absorbed reference substance and faeces were collected by stripping. Diets compounded from mixtures of these ingredients were then used to examine the possibility of predicting the digestibility of formulated diets.
Apparent digestibility of crude protein ranged from 79% to 90%, lipids from 83% to 95% and energy from 72% to 88% in the different ingredients. Apparent digestibility of carbohydrates was lower and ranged from 49% to 77%. Apparent digestibility of amino acids was higher than that of crude protein and differences were found among digestibilities of individual amino acids.
Tests conducted using five compound diets indicated that ingredient digestibility was additive for protein, amino acids, lipids and energy, whereas the digestibility of carbohydrates in the compound feeds was slightly lower than predicted.
Diets for Sparus aurata may thus be formulated on the basis of digestibility of individual ingredients.     

Preliminary evaluation of pea seed meal in diets for gilthead sea bream (Sparus aurata) juveniles

Juvenile gilthead sea bream with a mean initial body weight of 5 g were fed for 12 weeks with experimental diets containing 10% and 20% fishmeal protein (sole protein source in the control diet) replaced by processed pea seed meals. The processed pea seed meals were dehulled, defibred, extruded and microground pea seed meal (PSM1) or whole pea treated by infrared radiation and ground (PSM2). Apparent digestibility coefficients of the experimental diets were determined in a separate trial. At the end of the growth trial there were no significant differences in growth performance, feed utilization or whole-body composition among experimental groups. There were no differences in apparent protein digestibility among experimental groups (except for fish fed PSM1 at the lowest inclusion level). Both dry matter and energy digestibility of the diets, including PSM2 and with the highest inclusion level of PSM1, were significantly lower than those of the control diet. The results of this study suggest that pea seed meal may replace up to 20% fishmeal protein in diets for gilthead sea bream juveniles without affecting fish performance. Further studies should focus on technological treatments to increase utilization of pea seed meal carbohydrate, as both apparent dry matter and energy digestibility were affected by dietary inclusion level and by pea seed meal processing method.     

Fatty acid digestibility in gilthead sea bream fed diets containing native,re‐esterified or acid vegetable oils

Re‐esterified vegetable oils are obtained from a chemical esterification reaction between vegetable acid oils and glycerol. Due to their properties, it is expected that they have a higher nutritive value than their corresponding acid oils and a better digestibility than their native counterparts. The aim of this study was to determine the effect of re‐esterified oils with a different monoacylglycerol (MAG) and diacylglycerol (DAG) content, produced from palm or rapeseed, on fatty acid digestibility in gilthead sea bream (Sparus aurata). Triplicate groups of fish were fed nine experimental diets containing different oils during 28 days. For each source, four different types of oil were used: native, re‐esterified low or high in MAG and DAG and acid. A commercial fish oil was used for the control diet. Diets containing re‐esterified oils had better apparent digestibility coefficients (ADC) of total fatty acids than acid oil diets. Re‐esterified oils do not negatively affect apparent digestibility coefficients of fatty acids when compared to their corresponding native oils and could be incorporated as a source of energy in diets for gilthead sea bream. An improvement in digestibility compared to the native oil diet was only obtained in palm re‐esterified oil high in MAG and DAG.     

Incorporation and metabolism of 14C-labelled polyunsaturated fatty acids in juvenile gilthead sea bream Sparus aurata L. 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 PUFAs were investigated in juvenile gilthead sea bream, Sparus aurata. The results indicate that juvenile gilthead sea bream have only limited ability to convert CH PUFAs to C₂₀ and C₂₂ HUFAs in vivo. The data are consistent with the results from nutritional studies on larvae, postlarvae and fingerlings that have shown that gilthead sea bream require the provision of preformed eicosapentaenoic and docosahexaenoic acids in the diet. The impairment in the desaturase/elongase pathway was quantitatively and qualitatively similar to that found in turbot, Scophthalmus maximus, being at the level of the 5-desaturase. The low activity of 5-desaturase combined with the consistent finding that arachidonic acid is selectively retained in membrane phosphatidylinositol suggests that, in addition to eicosapentaenoic and docosahexaenoic acids, gilthead sea bream may also have a requirement for preformed arachidonic acid in the diet.Abbreviations AA 5,8,11,14-eicosapenaenoic acid (arachidonic acid, 20:4n–6) - CPL diradyl (diacyl + alkenylacyl + alkylacyl) glycerophosphocholine - DHA 4,7,10,13,16,19-docosahexaenoic acid (22:6n–3) - EPA 5,8,11,14,17-eicosapentaenoic acid (20:5n–3) - EPL diradyl (diacyl, alkenylacyl + alkylacyl) glycerophosphoethanolamine - HUFA highly unsaturated fatty acids ( C₂₀ and with 3 double bonds) - LA 9,12-octadecadienoic acid (linoleic acid, 18:2n–6) - LNA 9,12,15-octadecatrienoic acid (-linolenic acid, 18:3n–3) - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acid(s)     

Recovery of opercular anomalies in gilthead sea bream, Sparus aurata L.: morphological and morphometric analysis

Opercular anomalies are very frequent in reared gilthead sea bream and these can negatively influence the product value. Field observations have suggested that opercular malformations can recover over time. In order to verify this hypothesis, 140-day-old gilthead sea bream with monolateral opercular anomalies were divided into three groups, according to the type and increasing seriousness of the opercular malformations, and another group was composed of fish with bilateral opercular anomalies. All groups were monitored for 16 months. In the group with monolateral anomalies, the opercular recovery process was documented by morphological (stereomicroscope) and morphometric analysis. For the latter analysis, two relevant areas, A and T, were identified in the cephalic region. The ratio (T - A)/T, tending to 1, represents the recovery index (RI) of anatomical integrity and quantifies the recovery level of opercular complex anomalies. Results suggested that the recovery process was considerable over the 16 months of investigation but should not be considered complete. At the end of the study, 61% of the gilthead sea bream population with monolateral opercular defects recovered external integrity, whereas the population with bilateral defects showed a poorer recovery capability.     

Influence of dietary levels of soybean meal on the performance and gut histology of gilthead sea bream (Sparus aurata L.) and European sea bass (Dicentrarchus labrax L.)

The present study was performed to determine the effect of soybean meal (SBM) on the performance and gut histology of gilthead sea bream and European sea bass. Three isonitrogenous and isolipidic extruded diets (crude protein, 470 g kg⁻¹ diet; crude fat, 200 g kg⁻¹ diet) were formulated containing 0 (0 SBM), 180 (180 SBM) and 300 (300 SBM) g kg⁻¹ diet SBM and tested on both species in two separate experiments. Fish at an initial average weight of around 18 g were randomly allocated to 800 L square tanks connected to a closed recirculating system. The trials lasted 80 days for sea bream and 89 days for sea bass. Fish were hand-fed to apparent satiation. Increasing the level of SBM had no significant effects on the specific growth rate, feed intake and feed conversion rate in both the species. In the sea bream distal intestine, lamina propria was moderately and diffusely expanded in some fish due to an increase in cell infiltration represented by mononuclear cells, this finding being more frequent in animals fed diet 300 SBM. No other morphological alterations in intestinal folds, enterocytes or other inflammation signs were noticed in the sea bream distal intestine of any group. No histological differences were found in sea bass in any experimental group.