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.     

Characterization of nodavirus and viral encephalopathy and retinopathy in farmed turbot, Scophthalmus maximus (L.)

An outbreak of nodavirus infection in turbot larvae is described with respect to histopathology, immunohistochemistry, cell culture cultivation, RT-PCR amplification and sequence analysis of the capsid protein gene RNA2. Affected turbot developed classical signs of viral encephalopathy and retinopathy (VER) with abnormal swimming behaviour and high mortality levels. In the acute stage of infection, light microscopy revealed vacuolation of the central nervous system (CNS), with positive immunohistochemical staining for nodavirus. Later in the infection, CNS lesions appeared more chronic and contained clusters of cells immunopositive for nodavirus. Bacterial overgrowth in the intestines of the fish may have provoked or influenced the course of the nodavirus infection. We were unable to propagate the virus in cell culture. While RT-PCR using primers designed to detect Atlantic halibut nodavirus gave negative results, further testing with primers complementary to a more conserved region of RNA2 resulted in amplification of a product of the expected size. The entire RNA2 segment was cloned and sequenced. Sequence alignment showed that the turbot nodavirus (TNV) was different from previously described fish nodaviruses. In addition, phylogenetic analysis based on an 823 nt region of the sequence indicated that TNV clustered outside the four established fish nodavirus genotypes, suggesting a fifth genotype within the betanodaviruses.     

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。     

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.     

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.     

Effect of diet composition on nutrient digestibility and digestive enzyme levels of gilthead sea bream (Sparus aurata L.)

An experiment was conducted to evaluate the effects of dietary composition on apparent digestibility coefficients of protein, fat and carbohydrate of gilthead sea bream. In addition α‐amylase and proteolytic activities were measured in the digestive tract of fish, held at 20±1°C. Six experimental diets were formulated containing approximately 40%, 45% and 50% protein, 11% and 21% fat and a starch level which fluctuated from 14% to 36%. Fish having an average weight of 100–130 g were used, fed at 1.5% of their body weight daily, and digestibility was measured using an indirect method. Enzyme activities were measured in the digestive tract of fish, fed diets containing a combination of 40%, 50% protein with 11%, 21% fat at 0.5, 5, 10, 24 h after a single meal. Starch digestibility was reduced with its level in the diet. It also negatively affected fat digestibility as well as protein digestibility, the last only at the high fat level. Dietary fat level had a negative effect on starch and protein digestibility. Fat affected also strongly α‐amylase levels in the digestive tract, while its effect on protease activity was of smaller magnitude. These results indicate that digestive enzyme activities and nutrient digestibility values are affected by dietary composition, carbohydrates and fat indicating the strongest effect.     

Growth in adult gilthead sea bream (Sparus aurata L) as a result of interference in sexual maturation by different photoperiod regimes

Gilthead sea bream (Sparus aurata L) pass through the period of sexual maturation while being intensively reared for commercial purposes. This process affects growth and food conversion efficiency. In an attempt to reduce this problem several lighting regimes have been utilized which impede gonad development by increasing the length of the day. A four hour increase in natural day length three months before reaching sexual maturation in fish of 390 g, improves growth in comparison to the control by about 15% (means SD of 606.2 ± 10.6 and 526.5 ± 12.7 g, respectively) towards the end of the natural reproductive season. However, these differences are subsequently compensated since the sexual maturation process is only delayed (rather than being permanently halted). The manipulated groups reached maturity and then stopped growing between three and four months after the control groups. For this reason, it would appear advantageous to manipulate sexual maturation through photoperiod alteration in those groups which go to market either during or towards the end of the reproductive season since greater fish weights will be achieved in addition to an improved food efficiency.     

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.     

Effectiveness of four methods for tagging juveniles of farm-reared gilthead sea bream, Sparus aurata, L.

This study tested different tags for each size of farm-reared gilthead sea bream, Sparus aurata L. juveniles, used in enhancement programmes. The following tags were tested: dye mark, fingerling tag, opercular tag, anchor tag A (filament diameter of 1.20 mm) and anchor tag B (0.85 mm). Tag/mark retention, wounds produced by the tag, ease of application, ease of detection and cost were examined. Epidermal dye marks had poor retention and detection rates. Of the tags tested, retention was greater for the anchor tag A (80–100%) after 7 months; anchor tag B showed a retention rate of only 20% after 3 months. The fingerling tag had a retention rate of 27.5% after 12 months. The opercular tag had a retention rate of 44% after 20 days but fractured the opercular bone in many cases. Tag incisions and wounds persisted in some fish for the duration of the experiment.     

Amino acid retention of gilthead sea bream (Sparus aurata,L.) fed with pea protein concentrate

A trial of 218 days of duration was carried out to assess the use of pea protein concentrate (PPC) as a substitute for fish meal in diets for juvenile gilthead sea bream (52 g average initial weight), using four diets (0, 16, 32 and 48) with PPC inclusion levels of 0, 162, 325 and 487 g kg^?1, respectively. At the end of the trial, the fish reached weights of 397, 385, 383 and 355 g for 0, 16, 32 and 48 diets, respectively; diet 48 gave the lowest specific growth rate (SGR, 0.88% per day) but 0%, 16% and 32% PPC did not present statistical differences. Feed conversion ratio (FCR), protein efficiency ratio (PER) and body composition were not affected by the diets. Digestible protein retention was significantly higher with the 0% diet, but no energy retention differences were observed. Retention rates of ingested and digestible amino acids were very similar; in general, the retention of essential amino acids decreased as dietary PPC increased, and lysine retention was the highest and phenylalanine retention was the lowest. From the results of this experiment, it may be concluded that fish meal can be replaced up to 32% by PPC in sea bream without negative effects on fish weight gain, FCR and PER.     

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.     

The effects of long-day photoperiod on growth, body composition and skin colour in immature gilthead sea bream (Sparus aurata L.)

Immature gilthead sea bream (Sparus aurata L.) with a mean initial weight of 25.6 g were reared over 11 months to market size under different photoperiods: 16L:8D; 24L:0D and a control. Differences in final mean weight were significant between the three treatments (P<0.001): 16L:8D, 465.0 g; 24L:0D, 445.9 g; control, 402.6 g. Fish from the 24L:0D consumed most while 16L:8D fish converted it most efficiently. The lipid content of fillets was lower (P<0.05) for the 24L:0D (5.37±0.16 g 100 g^?1 of fillet) than for the control (6.02±0.12 g), and the moisture content lower for the control than for either treatment groups. Skin luminosity (L^*) was directly related to the number of hours of light exposure.     

Interactions of microorganisms isolated from gilthead sea bream, Sparus aurata L., on Vibrio harveyi, a pathogen of farmed Senegalese sole, Solea senegalensis (Kaup)

Four bacterial isolates from farmed gilthead sea bream, Sparus aurata, included in a previous study as members of the Vibrionaceae and Pseudomonodaceae and the genus Micrococcus, have been evaluated for their adhesive ability to skin and intestinal mucus of farmed Senegalese sole, Solea senegalensis, and their antagonistic effect on Vibrio harveyi, a pathogen of sole. These isolates showed higher adhesion to sole mucus than the pathogenic strains of V. harveyi assayed. Only two of the isolates showed antagonistic activity to V. harveyi. Interactions of the four isolates with V. harveyi in respect of adhesion to skin and intestinal mucus under exclusion, competition and displacement conditions were studied. Three isolates were able to reduce the attachment to skin and intestinal sole mucus of a pathogenic strain of V. harveyi under displacement and exclusion conditions, but not under competition conditions. The in vivo probiotic potential of isolate Pdp11 was assessed by oral administration followed by challenge with the pathogenic V. harveyi strain Lg14/00. A group of 50 Senegalese sole received a commercial diet supplemented with 10(8) cfu g(-1) of lyophilized Lg14/00 for 15 days. A second group of fish received a non-supplemented commercial diet. After challenge the mortality of the fish receiving the diet supplemented with the potential probiotic isolate was significantly lower than that in the fish receiving the non-supplemented commercial diet. This study has shown that the ability to interfere with attachment of pathogens, as well as the adhesion to host surfaces, are suitable criteria for selection of candidate probiotics for use in the culture of Senegalese sole.     

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.     

Optimum dietary soybean meal level for maximizing growth and nutrient utilization of on-growing gilthead sea bream (Sparus aurata)

Six isonitrogenous [450 g kg⁻¹ crude protein (CP)] and isoenergetic diets (23 kJ g⁻¹) with six levels of defatted soybean meal inclusion (0, 132, 263, 395, 526 and 658 g kg⁻¹) in substitution of fish meal were evaluated in gilthead sea bream of 242 g initial weight for 134 days. Fish fed diets S0, S13, S26 and S39 had a similar live weight (422, 422, 438 and 422 g, respectively) but fish fed diets S53 and S66 obtained the lowest final weight (385 and 333g, respectively), and similar results were presented in specific growth rate (SGR). Fish fed diets S53 and S66 also obtained the highest feed conversion ratio (FCR). Quadratic multiple regression equations were developed for SGR and FCR which were closely related to dietary soybean level. The optimum dietary soybean levels were 205 g kg⁻¹ for maximum SGR and 10 g kg⁻¹ for minimum FCR. Sensorial differences were appreciated by judges between fish fed S0 and S39 soybean level, but after a re-feeding period of 28 days with diet S0, these differences disappeared.     

Pharmacokinetics of ivermectin in sea bream, Sparus aurata using a direct competitive ELISA

The pharmacokinetics of ivermectin in the serum of cultured sea bream, Sparus aurata, after a single intraperitoneal injection of 100 μg kg⁻¹ body weight was studied by the use of direct competitive ELISA. Pharmacokinetic analysis of the serum concentrations versus time points obtained was performed using non-comparmental analysis and a compartmental pharmacokinetic model approach. In the latter case a two-compartment open model with a lag time gave the best fitting. The maximum peak serum concentration was 308.4 ng ml⁻¹ at 2 h post treatment. The AUC of ivermectin was 10700 ng h ml⁻¹ and the elimination half-life 15.37 h, indicating a rapid uptake, high bioavailability and fast elimination of the drug by sea bream. This revised version was published online in July 2006 with corrections to the Cover Date.     

A new tool for determining the optimum fish meal and vegetable meals in diets for maximizing the economic profitability of gilthead sea bream (Sparus aurata,L.) feeding

The economic profitability of fish meal replacement by soybean meal and pea‐rice concentrate mixture, was analysed considering the economic conversion ratio (ECR) and the economic profit index (EPI_st) as a proposal for evaluating the economic profitability in diets for sea bream. The optimal fish meal levels for minimizing the ECR were 33% (when soybean meal was included) and 14% (in the case of pea‐rice concentrate mixture) and for maximizing the EPI_st, optimal fish meal levels were 31% and 28.8%, respectively, considering the current market price of fish meal and vegetal ingredients. The EPI_st, seems to be a better index than the ECR to estimate diet profitability, because it considers weight increment, feed conversion ratio, diet cost and fish sale price. A sensibility analysis was conducted, considering variations in the price of fish meal and vegetal ingredients. Finally, several multiple regressions were developed for estimating the optimal level of fish meal with each vegetable ingredient and the expected value of the economic profit as a function of the market price of both fish meal and plant protein. When fish meal prices increase, the highest beneficial level of the inclusion fish meal and the highest economic profit would be achieved with pea‐rice mixture.