Effects of postmortem storage temperature on sea bass (Dicentrarchus labrax) muscle protein degradation: analysis by 2-D DIGE and MS

Storage conditions are known to be important for postmortem deterioration of fish muscle, and temperature is one of the factors with the strongest impact on this process. In order to shed light on the influence of temperature on the status of sea bass (Dicentrarchus labrax) muscle proteins during postmortem storage, a 2-D DIGE and mass spectrometry study was performed on fish kept at either 1 or 18°C for 5 days. As expected, the greatest alterations in sea bass filet protein composition were observed upon postmortem storage at 18°C, with distinct changes appearing in the 2-D protein profile after 5 days of storage at this temperature. In particular, degradation of the myofibrillar protein myosin heavy chain and of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase, among the most abundant muscle proteins, could be clearly observed upon storage at higher temperatures. Although to a lesser extent, however, several proteins were observed to vary in abundance also upon storage for 5 days at 1°C. In particular, one of the most interesting observations was the rapid and significant decrease in the abundance of nucleoside diphosphate kinase B and phosphoglycerate mutase 2, which was observed also at low storage temperatures and appeared to be temperature-independent. The results of this study offer new knowledge on changes occurring in sea bass muscle proteins during postmortem storage at different temperatures and provide indications on protein degradation trends that might be useful for monitoring freshness of fish and quality of storage conditions.     

Fatty acids in liver and muscle of farmed and wild sea bass (Dicentrarchus labrax L.)

Growth rate and assimilation of essential fatty acids were analysed in sea bass (Dicentrarchus labrax L.) fed different diets. Gas Chromatographie analyses of total lipids in commercial diets indicated excessive presence of saturated, mono and dienoic fatty acids, while the content of to 3 HUFA was very low. Fatty acid patterns of liver and white muscle farmed sea bass reflected the contents of the dietary lipids. The concentration of polyunsaturated fatty acids, particularly ω3 HUFA was significantly lower when compared with samples from the wild population. The atherogenic and fish lipid quality indices confirmed the higher nutritional quality of wild sea bass when compared with farmed fishes.     

Zinc and copper in fish from natural waters and rearing ponds in Northern Italy

Zinc and copper were detected in several tissues of fresh and saltwater fish. Liver concentrations varied widely, with respect to the storage and detoxication functions of the organ. In muscular tissues the two metals are linked to aerobic metabolism being higher in the heart and lower in the white muscle. High levels of zinc were found in the female gonad, while in the brain zinc has been shown to be more constant and possibly regulated better than copper. In sea bass supplemented with artificial diets no correlation was found between the metal content in the diet and that of the tissue. In goldfish attempts using gel filtration to isolate specific metal binding proteins of low molecular weight gave negative results, the metals were mostly bound to ligands excluded from the gel.     

Survival and changes in growth of juvenile tench (Tinca tinca L.) fed defined diets commonly used to culture non-cyprinid species

The study is an attempt to evaluate the feasibility of intensive tench culture using non‐specific diets as a preliminary step to check the acclimatization of the species under intensive rearing systems. Five‐month‐old juvenile tench were reared in recirculating systems at mean water temperatures of 22°C for 75 days. Fish fed with four different commercial diets (trout starter, trout first feeding, sea‐bass and eel), showed significantly higher final weights than the fish fed either eel or sea‐bass diets. Initial weight for all treatments was 2.3 ± 0.53 g. Final weight for the commercial diet groups was 3.56 ± 0.4 g, compared with the remaining groups that reached 2.09 ± 0.47 g (P < 0.05). Significantly higher survival rates were observed in the eel and sea‐bass groups (84.7 and 51.5%, respectively) than in either of the trout diet groups (38%). Specific growth rates (1.26 vs –0.18) and condition factor (1.26 vs 0.93) were also higher than those fed with salmonid diets (P < 0.05). Results obtained in this study indicate that regardless of the species’ slow growth, when compared with other cyprinids, final growth rates and survival of tench fed exclusively on sea‐bass or eel diets can be considered satisfactory. It must be pointed out that these promising results were obtained at lower temperatures than previous studies of tench in culture systems. The use of belt feeders did not show improvement in growth compared with manually fed fish when trout diets were used.     

Development of leptin resistance in rat soleus muscle in response to high-fat diets

Direct evidence for leptin resistance in peripheral tissues such as skeletal muscle does not exist. Therefore, we investigated the effects of different high-fat diets on lipid metabolism in isolated rat soleus muscle and specifically explored whether leptin's stimulatory effects on muscle lipid metabolism would be reduced after exposure to high-fat diets. Control (Cont, 12% kcal fat) and high-fat [60% kcal safflower oil (n-6) (HF-Saff); 48% kcal safflower oil plus 12% fish oil (n-3)] diets were fed to rats for 4 wk. After the dietary treatments, muscle lipid turnover and oxidation in the presence and absence of leptin was measured using pulse-chase procedures in incubated resting soleus muscle. In the absence of leptin, phospholipid, diacylglycerol, and triacylglycerol (TG) turnover were unaffected by the high-fat diets, but exogenous palmitate oxidation was significantly increased in the HF-Saff group. In Cont rats, leptin increased exogenous palmitate oxidation (21.4 +/- 5.7 vs. 11.9 +/- 1.61 nmol/g, P = 0.019) and TG breakdown (39.8 +/- 5.6 vs. 27.0 +/- 5.2 nmol/g, P = 0.043) and decreased TG esterification (132.5 +/- 14.6 vs. 177.7 +/- 29.6 nmol/g, P = 0.043). However, in both high-fat groups, the stimulatory effect of leptin on muscle lipid oxidation and hydrolysis was eliminated. Partial substitution of fish oil resulted only in the restoration of leptin's inhibition of TG esterification. Thus we hypothesize that, during the development of obesity, skeletal muscle becomes resistant to the effects of leptin, resulting in the accumulation of intramuscular TG. This may be an important initiating step in the development of insulin resistance common in obesity.     

复合乳酸菌对冷藏海鲈鱼块的保鲜效果

【目的】研究复合乳酸菌对冷藏海鲈鱼块的保鲜效果。【方法】以冷藏海鲈鱼块为对象,筛选出3株能够明显抑制其优势腐败菌(草莓假单胞菌Pseudomonas fragi,腐败希瓦氏菌Shewanella putrefacens)生长的单一乳酸菌,同时也筛选出对其优势腐败菌具有最显著抑制效果的一组复合乳酸菌,再将该复合乳酸菌接种到海鲈鱼块上,在4°C冷藏过程中,通过感官评定、挥发性盐基氮(TVB-N值)的测定和优势腐败菌的计数来评价复合乳酸菌对冷藏海鲈鱼块的保鲜效果。【结果】单一乳酸菌(干酪乳杆菌LC1、植物乳杆菌LP1和乳酸菌L3)对2株冷藏海鲈鱼优势腐败菌的抑制效果明显;复合乳酸菌(干酪乳杆菌LC1+植物乳杆菌LP1+乳酸菌L3)的抑菌效果最为显著;在4°C冷藏过程中,复合乳酸菌能使冷藏海鲈鱼块发生感官变化延缓6 d、使TVB-N值的升高延缓2 d,同时显著抑制优势腐败菌的生长。【结论】复合乳酸菌对冷藏海鲈鱼块具有良好的保鲜作用,能有效延长其货架期。     

Modelling swimming activities and energetic costs in European sea bass (Dicentrarchus labrax L., 1758) during critical swimming tests

Muscular activity patterns in red and white muscles linked to oxygen consumption were studied during critical swimming tests in the sea bass (Dicentrarchus labrax Linnaeus 1758). The species is one of the most important for Mediterranean Sea aquaculture. A sigmoid model was used to fit both the oxygen consumption and red muscle activity while the white muscle activity pattern was described by an exponential model. Red muscle reaches an activation plateau close to critical swimming speed mostly due to the oxygen diffusion velocity in tissues. The exponential activation of white muscle appears to be linked to short and sudden periods of great energy need to cope with adverse conditions such as predation and escape. Both oxygen consumption and muscular activity were found to be size dependent. The bioenergetics of sea bass was modelled based on fish mass and swimming speed to predict the minimum and maximum speed as well as the mass-specific active metabolic rate and standard metabolic rate. An important finding was that contrary to other well-known species, swimming at subcritical speeds in sea bass involves both red and white muscle in different proportions.     

Fillet yield and chemical composition of farm-raised sunshine bass (Morone chrysops♀ × Morone saxatilis♂) fed high-energy diets

Fillet yield and chemical composition were evaluated in small, medium and large (about 150, 350 and 700 g, respectively) size classes of sunshine bass ( Morone chrysops ♀ ×  Morone saxatilis ♂) fed high-energy diets. Fish size had a major effect on the fillet yield, which was significantly higher (38.8 vs. 33.4–34.6%) in fish belonging to the medium-size class. The lipid contents of both whole and skinned fillets were higher than those reported in the literature, ranging from 7.8 to 10.6% in whole fillets and from 5.9 to 7.6% in skinned fillets, thereby allowing classification of sunshine bass fed high-energy diets as a medium-fat fish. A significant effect of fish size on lipid content was found only for whole fillets, therefore indicating that dietary energy excess in larger fish is mainly stored as subdermal fat. The dietary fatty acid profile was found to be the major determinant of edible muscle lipid composition in all size-classes considered. The cholesterol content of muscle tissue ranged from 23 to 29 mg/100 g tissue.     

Influence of dietary composition on growth and energy reserves in tench (Tinca tinca)

The effects of different protein, lipid and carbohydrate diets on growth and energy storage in tench, Tinca tinca L., were studied. Over a 2-month period fish were fed four different diets: control, protein-enriched, carbohydrate-enriched and lipid-enriched. The best growth rates were obtained with the control and protein-enriched diets; the carbohydrate diet produced the worst results (lowest specific growth rate, weight gain, nutritional index and hepatosomatic index). These results suggest that it is not advisable to reduce dietary fish protein below 35%, and that it is not possible to obtain a protein-sparing effect of either lipids or carbohydrates, at least in our experimental conditions. The high-protein diet resulted in the storage of energy excess as muscle proteins and hepatic glycogen. Tench fed the high-carbohydrate diet stored carbohydrates as muscle glycogen and reduced plasma triglycerides. Finally, both liver and muscle lipid content were in positive correlation to dietary lipid.     

在低蛋白质饲料中补充必需氨基酸对大口黑鲈生长、体组成和免疫指标的影响

为了研究在低蛋白质饲料中补充晶体必需氨基酸对大口黑鲈生长、体组成和免疫指标的影响,根据鱼体的必需氨基酸组成模式设计了7种等能的试验饲料。其中4种饲料（45CP、40CP、35CP和30CP）的粗蛋白质水平分别为45%、40%、35%和30%,另3种饲料（40AA、35AA和30AA）是在低蛋白质饲料（40CP、35CP和30CP）的基础上添加必需氨基酸,使它们的必需氨基酸含量与45CP（对照组）相一致。用上述饲料对初始体重为（10.13 0.01） g的大口黑鲈进行了89d的饲养试验。饲养试验在室内循环水养殖系统中进行,每种饲料设3个重复,每重复放养30尾鱼。方差分析显示:试验鱼的生长性能、饲料效率、全鱼和肌肉的粗蛋白质含量、成活率以及免疫指标均随着饲料蛋白质含量的降低而显著降低（P0.05）。添加必需氨基酸的35AA和30AA的饲料效率和蛋白质保留率分别显著高于对应的未添加必需氨基酸的35CP和30CP组（P0.05）,但仍显著低于45CP组（P0.05）。40AA的试验鱼血清溶菌酶活性和血清补体活性与45CP组差异不显著（P0.05）。35AA和30AA组的头肾白细胞呼吸爆发活性显著高于35CP和30CP组（P0.05）。30AA组的全鱼粗蛋白质含量以及肥满度显著高于30CP（P0.05）。各组试验鱼的水分和灰分均无显著差异（P0.05）。回归分析显示:在低蛋白质饲料中补充晶体必需氨基酸对大口黑鲈幼鱼的生长、饲料效率和蛋白质保留率所产生的影响与其引起增加了的饲料蛋白质水平而不是饲料的必需氨基酸水平正相关。研究表明,在低蛋白质饲料中补充的晶体必需氨基酸对大口黑鲈的生长、体组成和免疫指标产生的有益作用不及等量的以蛋白质为来源的必需氨基酸。       

Self-design of fish diets by means of self-feeders: validation of procedures

In this paper, we show the results from four different experiments in which sea bass, maintained under laboratory conditions, could choose between two or three different diets through self-feeders, which gradually increased the complexity and potential range of selection, to design their own diet in accordance with their requirements. At first, sea bass were allowed to select between two complete diets differing in the proportion of protein (52-58%): this showed their capacity to distinguish between two diets made of the same ingredients. Next, two incomplete diets, containing a fixed amount of protein (56%) and lacking either fat or carbohydrate, were made available. Three mixed diets made up of pairs of macronutrients (protein-carbohydrate, protein-fat or fat-carbohydrate) were tested in the next experiment and, finally, three diets containing only one macronutrient (protein fat or carbohydrate) were made available to fish. Taking into account selection made by the fish in the first three experiments, in which macronutrient selection was statistically different, protein was the main macronutrient chosen by fish (278.15 kJ/kgBW/day, on average), followed by fat and carbohydrate (162.85 and 64.56 kJ/kgBW/day, respectively). In conclusion, the results reveal the ability of sea bass to select an appropriate diet from experimental diets containing two or three macronutrients and suggest that the proposed methodology is a powerful tool for studying the differing nutritional needs of different species of fish.     

Prevention of high-fat diet-induced muscular lipid accumulation in rats by �� lipoic acid is not mediated by AMPK activation

Skeletal muscle triglyceride accumulation is associated with insulin resistance in obesity. Recently, it has been suggested that α lipoic acid (ALA) improves insulin sensitivity by lowering triglyceride accumulation in nonadipose tissues via activation of skeletal muscle AMP-activated protein kinase (AMPK). We examined whether chronic ALA supplementation prevents muscular lipid accumulation that is associated with high-fat diets via activation of AMPK. In addition, we tested if ALA supplementation was able to improve insulin sensitivity in rats fed low- and high-fat diets (LFD, HFD). Supplementing male Wistar rats with 0.5% ALA for 8 weeks significantly reduced body weight, both on LFD and HFD (−24% LFD+ALA vs. LFD, P < 0.01, and −29% HFD+ALA vs. HFD, P < 0.001). Oil red O lipid staining revealed a 3-fold higher lipid content in skeletal muscle after HFD compared with LFD and ALA-supplemented groups (P < 0.05). ALA improved whole body glucose tolerance (∼20% lower total area under the curve (AUC) in ALA supplemented groups vs. controls, P < 0.05). These effects were not mediated by increased muscular AMPK activation or ALA-induced improvement of muscular insulin sensitivity. To conclude, the prevention of HFD-induced muscular lipid accumulation and the improved whole body glucose tolerance are likely secondary effects due to the anorexic nature of ALA.     

Metabolic flexibility and insulin resistance

Metabolic flexibility is the capacity for the organism to adapt fuel oxidation to fuel availability. The inability to modify fuel oxidation in response to changes in nutrient availability has been implicated in the accumulation of intramyocellular lipid and insulin resistance. The metabolic flexibility assessed by the ability to switch from fat to carbohydrate oxidation is usually impaired during a hyperinsulinemic clamp in insulin-resistant subjects; however, this "metabolic inflexibility" is mostly the consequence of impaired cellular glucose uptake. Indeed, after controlling for insulin-stimulated glucose disposal rate (amount of glucose available for oxidation), metabolic flexibility is not altered in obesity regardless of the presence of type 2 diabetes. To understand how intramyocellular lipids accumulate and cause insulin resistance, the assessment of metabolic flexibility to high-fat diets is more relevant than metabolic flexibility during a hyperinsulinemic clamp. An impaired capacity to upregulate muscle lipid oxidation in the face of high lipid supply may lead to increased muscle fat accumulation and insulin resistance. Surprisingly, very few studies have investigated the response to high-fat diets. In this review, we discuss the role of glucose disposal rate, adipose tissue lipid storage, and mitochondrial function on metabolic flexibility. Additionally, we emphasize the bias of using the change in respiratory quotient to calculate metabolic flexibility and propose novel approaches to assess metabolic flexibility. On the basis of current evidence, one cannot conclude that impaired metabolic flexibility is responsible for the accumulation of intramyocellular lipid and insulin resistance. We propose to study metabolic flexibility in response to high-fat diets in individuals having contrasting degree of insulin sensitivity and/or mitochondrial characteristics.     

Effects of dietary protein and lipid levels on growth and feed utilization of wild‐caught striped sea bream,Lithognathus mormyrus

A feeding trial was carried out to determine the effects of dietary protein and lipid levels on the growth performance and feed utilization of wild‐caught striped sea bream (Lithognathus mormyrus). The experimental fish were collected from a local lagoon (Çardak Lagoon, Çanakkale, Turkey), transferred to the Marine Net Cage Unit and fed by hand to apparent satiation with a commercial sea bream feed (Biomar; 42% crude protein, 16% crude lipid). Approximately 4 weeks were needed to acclimate the fish to farming conditions. No pathological signs were observed and no fish losses occurred during the adaptation period. For the test trials four test diets with different levels of protein and lipid were formulated [low protein and low lipid (LP:LL), low protein and high lipid (LP:HL), high protein and low lipid (HP:LL), and high protein and high lipid (HP:HL)] and fed to L. mormyrus (mean weight 85.0 ± 4.6 g SEM) in the net cages (Ø 2 m, depth 2.5 m) for 60 days. During the experiment water temperature varied between 21.1 and 26.4°C; dissolved oxygen 8.4–9.6 mg L^?1; pH 7.2–8.6; and salinity 23.3–25.6‰. Growth performances of fish fed high protein diets were higher compared to fish fed low protein diets, irrespective of the dietary lipid level (P < 0.05). Feed conversion ratio (FCR) and protein efficiency ratio (PER) were not influenced by dietary protein or lipid levels (P > 0.05). Preliminary results indicate that striped sea bream can be easily adapted to farming conditions in net cages, and that a diet containing 50% crude protein and 15% crude lipid (HP:LL) levels with 23.0 g protein MJ^?1 gross energy of protein/energy ratio would be suitable for striped sea bream growth.     

Tryptophan requirement of juvenile Asian sea bass Lates calcarifer

The dietary requirement of tryptophan for juvenile Asian sea bass (Lates calcarifer Bloch) was studied. The juveniles (mean initial weight, 5.30 ± 0.06 g) were given semi‐purified test diets containing fish meal, gelatin, squid meal, and crystalline amino acids, for 12 weeks. Each set of isonitrogenous and isocaloric test diets contained graded levels of tryptophan. Fish (15 per tank) were reared in 250‐L fiberglass tanks provided with continuous flow‐through sea water at 26°C and salinity of 28 p.p.t. Fish were fed twice daily at a feeding rate of 8% of the body weight day^?1 for the first 4 weeks and at 3.5–2.5% of the body weight day^?1 from 5 to 12 weeks. The experiment was in a completely randomized design with two replicates per treatment. Mean percentage weight gains and feed efficiency ratios were significantly different in fish fed varying tryptophan levels. Survival was 100% in all treatments. On the basis of break‐point analysis of the growth response, the dietary tryptophan requirement of juvenile Asian sea bass is 0.41% of the dietary protein. This information will be useful in further refinement of practical feed formulations for the Asian sea bass.     

Synchronizing Effect of Photoperiod on the Dual Phasing of Demand-Feeding Rhythms in Sea Bass

Under natural environmental conditions, sea bass feeding rhythms are nocturnal in winter and diurnal the rest of the year. In this paper we describe the effect of contracting and expanding photoperiods and two skeleton photoperiods (SP) on the dual feeding rhythms of sea bass ( Dicentrarchus labrax L. ). To this end, twelve animals were held individually with access to self-feeders. First, the lights on and lights off were progressively delayed and advanced respectively by one hour in group 1 (G1), and conversely in group 2 (G2), so that the fish were exposed from a light/dark (LD) 12L:12D cycle to 2:22 LD (G1) and DL (G2) cycles and finally 0.25:23.75 LD (G1) and DL (G2). In the second experiment two SP's were used involving two light pulses separated by 12 hours, each pulse lasting 0.25 hours during the first two weeks and one hour during the succeeding two weeks. The results showed that diurnal and nocturnal sea bass tended to confine their feeding phase following the contraction of the LD cycle. Both SP's failed to simulate a complete photoperiod. In conclusion, the LD cycle appeared to drive the daily feeding rhythms but, the photoperiod length did not itself control the inversions of nocturnal and diurnal fish, so that other factors, in addition to photoperiod, may be involved in the control of the annual rhythms of phase inversions in sea bass.     

Diel variation in diet composition of a riverine fish community

Although diel food habit studies have been undertaken on a number of individual species, few studies have examined diel variation in the diets of fish communities. We examined the diel diet variation and feeding periodicity of a fish community in the Juniata River, Pennsylvania. Nine species, totalling 1,098 fish, were collected at 4-h intervals over a 24-h period in October 1989, in numbers sufficient to describe their diel variation in diet composition. Diel variation in diet composition was evident in all species, as no single prey taxon was dominant in the diet of any species during any 4-h interval. Ephemeropterans were the most important prey taxa for four species of centrarchids, whereas chironomids were the main prey of banded killifish, mimic shiners, and spotfin shiners. Algae was the major component in the diet of spottail shiners, whereas bluntnose minnows contained mostly detritus. Feeding activities of rock bass, redbreast sunfish, and pumpkinseed occurred at low levels throughout the day; peak feeding occurred from 2000 to 0400 hours. Food consumption of smallmouth bass increased throughout the day with peak consumption occurring at 2000 hours. Non-centrarchids fed little during daylight hours and showed peak activity at 2000–2400 h. Construction of a 24-hour diet from six 4-h interval estimates and feeding periodicity data provided a comprehensive representation of the diel feeding ecology of all species collected.     

Lysine and arginine requirements of juvenile Asian sea bass (Lates calcarifer)

Two separate experiments were conducted to determine the dietary requirements of juvenile Asian sea bass Lates calcarifer Bloch for lysine and arginine. Fish (average initial weight: lysine experiment, 13.12 ± 0.12 g; arginine experiment, 2.56 ± 0.13 g) were given amino acid test diets for 12 weeks containing fish meal, zein, squid meal, and crystalline amino acids. Each set of isonitrogenous and isocaloric test diets contained graded levels of L ‐lysine or L ‐arginine. The feeding rate in the lysine experiment was at 4–2.5% of the body weight day^?1, while in the arginine experiment it was at 10–4% of the body weight day^?1. The fish (20 per tank, lysine experiment; 15 per tank, arginine experiment) were reared in 500‐L fibreglass tanks with continuous flowthrough sea water at 27 °C and salinity of 31 ppt in the lysine experiment and at 29 °C and salinity of 29 ppt in the arginine experiment. The experiments were in a completely randomized design with two replicates per treatment. Survival was high in fish given adequate lysine or arginine. Mean percentage weight gains were significantly different in fish fed varying levels of lysine or arginine. Fish fed high levels of L ‐arginine suffered high mortalities. No significant differences were obtained in the feed efficiency ratios (FER, g gain g^?1 feed) of fish fed graded lysine, although the values tended to increase as the dietary lysine level was increased up to the requirement level. In contrast, in the arginine experiment, significant differences in FER of fish among treatments were obtained; the highest FER was observed in fish fed the diet containing an optimum arginine level. On the basis of the growth response, survival, and FER, the lysine and arginine requirements of juvenile Asian sea bass were estimated to be 20.6 g kg^?1 dry diet (4.5% protein) and 18.2 g kg^?1 dry diet (3.8% protein), respectively. These data will be useful in the further refinement of practical diet formulations for the Asian sea bass.