Assessment of reference dietary amino acid pattern for juvenile red sea bream, <Emphasis Type="Italic">Pagrus major</Emphasis>

Four semi-purified diets, containing crystalline amino acids (CAAs), were fed to juvenile red sea bream, Pagrus major in order to ascertain the ideal dietary amino acid pattern for this species. A control diet containing 50% casein–gelatin as protein sources, but no CAAs were fed to the fish. The other diets contained 30% casein–gelatin and 20% CAAs. CAAs were added to diets to simulate with amino acid pattern of the red sea bream eggs protein (REP), red sea bream larvae whole body protein (RLP), red sea bream juvenile whole body protein (RJP), and brown fishmeal protein (BFP). The juveniles (average initial body weight, 1.58 ± 0.01 g) were maintained in triplicate tanks and fed twice daily for 30 days. The highest weight gain was observed in juveniles fed the RJP diet. No significant difference was observed in juveniles fed the RLP and BFP diet. Feed efficiency ratio, protein efficiency ratio and amino acid retention in the whole body were significantly (p < 0.05) affected by the simulated dietary amino acid patterns. The essential amino acid profile and A/E ratios of the whole body after the growth trial showed little difference among the dietary treatments. The results suggest that red sea bream juveniles are able to utilize high amounts of CAA in coated form. The amino acid pattern of RJP could be used as an appropriate of reference dietary amino acid for this species.     

Effect of dietary medicinal herbs on lipid metabolism and stress recovery in red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

The effect of dietary medicinal herbs on lipid metabolism and stress recovery was investigated in red sea bream Pagrus major. Fish (mean body weight 24.0 ± 0.2 g) were fed on test fish meal diets supplemented with either Massa Medicata (Mm), Crataegi Fructus (Cf), Artemisia capillaries (Ac), or Cnidium officinale (Co), or with a mixture of the four herbs (HM) for 12 weeks. A control group was fed a diet without herbs in the same manner. A high survival rate was observed in the herbal diet groups. The final mean body weight, feed efficiency, protein efficiency ratio, and apparent protein and lipid retention in the Ac, Co, and HM groups were higher than those in the control and Mm groups. Final carcass, hepatic lipid and triglyceride contents, and plasma triglyceride and nonesterified fatty acid levels were lower in the Ac, Co, and HM groups compared to those of control and Mm groups. However, final hepatic phospholipid, plasma phospholipid, and high density lipoprotein cholesterol levels were higher in the Ac, Co, and HM diets groups than in the control and Mm groups. The Cf, Co, Ac, and HM groups showed faster recovery time in the 2-phenoxyethanol anesthesia test and a higher recovery rate in the 10-min air exposure test than those of the control and Mm groups. Moreover, the Cf, Ac, Co, and HM diet groups had a significantly lower plasma cortisol level than the control and Mm diet group, but the glucose level in the herbal diet groups was higher than that in the control group after a 1-h air exposure. These results indicate that the addition of medicinal herbs to the fish diet improved lipid utilization and stress recovery in red sea bream.     

Effect of the squid viscera hydrolysate on growth performance and digestion in the red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

The improvement in feed efficiency is one of the most important subjects in fish culture. The development of feed, in terms of good intake, high growth performance, and high feed efficiency is needed. Squid viscera are one of the candidates for alternative material in improving feed efficiency in fish culture. In the present study, we described the dietary effect of the squid viscera hydrolysate (SVH) on the growth performance of the red sea bream. The addition of SVH to feed caused significant increases in feed intake, fork length, and body weight and produced a marked improvement in feed conversion after 4 weeks of feeding. Furthermore, the results of this feeding revealed that low dietary levels of SVH promote growth performance in the red sea bream. We physiologically analyzed digestion and appetite in fish fed diet containing SVH. SVH promoted the activity of hepatic trypsin and lipase, gene expression of stomach pepsin, hepatic lipase, and pyloric caeca trypsin, thereby improving the nutrient availability in red sea bream. Moreover, the mRNA expression of appetite regulating factor, such as brain NPY and stomach ghrelin was significantly improved by dietary SVH. Our current results indicate that dietary SVH as alternative material produced excellent effects on growth performance, which is dependent on the promoting effect on digestion and appetite in red sea bream.     

Effect of water temperature and light intensity on swim bladder inflation and growth of red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis> larvae

This study aimed to examine the effect of different water temperatures and light intensities on swim bladder inflation (SBI) and growth of red sea bream Pagrus major larvae to improve rearing techniques for this species. Two sets of experiments were conducted: different rearing temperatures were used in experiment 1 (19, 21, 23, and 25 °C), and different light intensities in experiment 2 (250, 1000, 4000, and 16,000 lx). Water temperature did not affect SBI frequency, but SBI initiation was accelerated at higher temperature, i.e., it was initiated on 3 days post-hatching (dph) at 25 °C and on 6 dph at 19 °C, suggesting that the promotion period for SBI, which needs a surface skimmer to be  run, also accelerated with increasing temperature in red sea bream larviculture. A higher temperature also significantly promoted larval growth, although the notochord of larvae at SBI initiation was shorter at higher temperatures. Light intensity had no effect on either the initiation or the frequency of SBI. However, light intensity of 250 lx significantly reduced early larval growth compared to light intensities higher than 1000 lx. These results indicate that light at an intensity greater than 1000 lx at the water surface is suitable for the early larviculture of red sea bream.     

Use of acceleration loggers in aquaculture to determine net-cage use and field metabolic rates in red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

We investigated the usefulness of acceleration loggers in aquaculture by examining net-cage use and metabolic rates in red sea bream, Pagrus major. First, the fish’s metabolic rate (mg O₂ kg⁻¹ min⁻¹) was measured with the logger in a swim tunnel at designated water velocities. We found that metabolic rate could be expressed by using a linear regression model of the activity rate index (unitless min⁻¹) derived from acceleration data. Using this equation, the field metabolic rates of three fish in a net cage were monitored and were estimated at 14.1–15.0 kcal kg⁻¹ day⁻¹. The results suggested that 15–19% of energy from satiation feeding ration was consumed for metabolism and activity in the net cage. The loggers showed orderly net-cage use by the fish. Tagged individuals used the whole cage from surface to bottom, but individual fish that preferred the surface area rarely used the bottom, and vice versa. Metabolic rate increased significantly with distance of the fish from their preferred depths. The logger provided information on the physiological and behavioral responses of fish in a given breeding system, and its use should contribute to the design of practical aquaculture systems.     

Physiological response,blood chemistry profile and mucus secretion of red sea bream (<Emphasis Type="Italic">Pagrus major</Emphasis>) fed diets supplemented with <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> under low salinity stress

Environmental stressors caused by inadequate aquaculture management strategies suppress the immune response of fish and make them more susceptible to diseases. Therefore, efforts have been made to relieve stress in fish by using various functional feed additives in the diet, including probiotics. The present work evaluates the effects of Lactobacillus rhamnosus (LR) on physiological stress response, blood chemistry and mucus secretion of red sea bream (Pagrus major) under low salinity stress. Fish were fed four diets supplemented with LR at [0 (LR0), 1 × 10² (LR1), 1 × 10⁴ (LR2) and 1 × 10⁶ (LR3) cells g^?1] for 56 days. Before stress, blood cortisol, urea nitrogen (BUN) and total bilirubin (T-BIL) showed no significant difference (P > 0.05), whereas plasma glucose and triglyceride (TG) of fish-fed LR2 and LR3 diets were significantly lower (P < 0.05) than those of the other groups. Plasma total cholesterol (T-CHO) of fish-fed LR3 diet was significantly (P < 0.05) lower than that of the other groups. Furthermore, total plasma protein, mucus myeloperoxidase activity and the amount of mucus secretion were significantly enhanced in LR-supplemented groups when compared with the control group (P < 0.05). After the application of the low salinity stress test, plasma cortisol, glucose, T-CHO and TG contents in all groups showed an increased trend significantly (P < 0.01) compared to the fish before the stress challenge. However, plasma total protein and the amount of secreted mucus showed a decreased trend in all groups. On the other hand, BUN, T-BIL and mucus myeloperoxidase activity showed no significant difference after exposure to the low salinity stress (P > 0.05). In addition, the fish that received LR-supplemented diets showed significantly higher tolerance against low salinity stress than the fish-fed LR-free diet (P < 0.05). The physiological status and the detected immune responses, including total plasma protein and mucus myeloperoxidase activity in red sea bream, will provide a more comprehensive outlook of the effects of probiotics to relieve stress in fish.     

Use of <Emphasis Type="Italic">Porphyra</Emphasis> spheroplasts as feed additive for red sea bream

ABSTRACT:   Two test diets with and without 5% Porphyra spheroplasts (PS) were formulated using white fishmeal as the main protein source. Red sea bream Pagrus major (mean body weight 15.4 ± 0.1 g) were maintained in a flow-through system (100 L) of thermo-controlled sea water (salinity 32–34, 25°C, 8 L/min) with ordinary aeration (400–600 mL/min) under laboratory light conditions (light–dark 12 h:12 h). Fish were fed three times a day at 10:00, 14:00 and 18:00 hours by hand for 42 days at 6% body weight on each experimental diet. Studies revealed that growth performance, survival and nutrient retention were significantly ( P  < 0.05) higher in the groups fed a diet containing spheroplasts (PS diet). Further, the fish fed the PS diet showed significantly ( P  < 0.05) lower feed conversion rates. Both groups of the PS and control diets had similar levels of body nutritional profile in terms of proximate compositions and fatty acids without compromising blood serum related parameters. From these experimental results, thus, it is comprehensible that a supplementary diet containing Porphyra spheroplasts can be used for maximizing not only growth of P. major but also for utilization of the feed ingredients.     

Effect of taurine supplementation on hepatic metabolism and alleviation of cadmium toxicity and bioaccumulation in a marine teleost,red sea bream, <Emphasis Type="Italic">Pagrus major</Emphasis>

This study was performed to unravel the mechanism of the beneficial action of taurine on marine teleost fish, red sea bream (Pagrus major), by analyzing the hepatic metabolism. Moreover, the ameliorative effects of the nutrient against cadmium toxicity and bioaccumulation were further evaluated. The fish were fed a diet containing 0 % (TAU0 %), 0.5 % (TAU0.5 %), or 5.0 % (TAU5.0 %) taurine for 40–55 days (d) and subjected to cadmium acute toxicity and bioaccumulation tests. Taurine deficiency in feed severely affected growth and the hepatic metabolic profiles of the fish, including a remarkable increase in myo-inositol, aspartate, and ß-alanine in the TAU0 % group, which indicates a complementary physiological response to taurine deficiency. For the acute toxicity test, fish were fed the test diets for 55 d and were then exposed to different dose of cadmium ranging from 0 to 5.6 mg/L for 96 h. Fish fed taurine had a higher tolerance to cadmium than those not fed taurine. For the bioaccumulation test, fish were fed the test diets for 40 d and then were chronically exposed to 0.2 mg/L of cadmium for 28 d followed by depuration for 21 d. Cadmium concentrations in the liver and muscle of fish fed TAU5.0 % were significantly lower than those of fish fed TAU0 % for the first 7 d of exposure and the first 7 d of elimination. Our findings suggest a possible mechanism for the beneficial role played by taurine and that the inclusion of taurine in fish aquaculture feed may reduce cadmium contamination of fish intended for human consumption.     

Effects of organic acids on growth and phosphorus utilization in red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

ABSTRACT:   A feeding experiment was conducted to investigate the effects of various organic acids and supplements on the growth, phosphorus (P) utilization, and environmental loading of nitrogen (N) and P in red sea bream Pagrus major. Seven diets designated as PA (0.5% inorganic P as Ca[H₂PO₄]₂), PO (no inorganic P), CA (1% citric acid), MA (1% malic acid), LA (1% lactic acid), MHA (1% methionine hydroxy analog) and LTE (1% liquid trace elements) were formulated. Duplicate groups of 25 fish were fed four times daily to near satiation for 75 days. Fish fed CA and LTE diets showed significantly better weight gain and feed conversion ratio compared to fish fed diet PO. Absorption of P was significantly improved in all organic acid-supplemented groups compared to the PA and PO groups. The P retention in CA and LTE groups were significantly higher compared to PA and PO diets. The P excretion was also significantly decreased in all organic acid-supplemented groups. Thus, use of organic acids, in particular CA and LTE, in red sea bream diets can reduce the use of inorganic phosphorus and contribute to the development of much-desired eco-friendly diets.     

Dietary medicinal herbs improve growth and some non-specific immunity of red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

The effects of dietary medicinal herbs on growth and some non-specific immunity were investigated in juvenile red sea bream Pagrus major. The fish (mean body weight 24.0±0.2g) were fed fishmeal diets supplemented with either Massa medicata (Mm), Crataegi fructus (Cf), Artemisia capillaries (Ac), Cnidium officinale (Co), or a mixture of all the herbs (HM), and a control diet without medicinal herbs, for 12 weeks. Survival, specific growth rate, feed efficiency, condition factor and hemoglobin levels were higher in fish given herbal diets than fish given the control diet without herbs. Significantly higher serum high density lipoprotein-cholesterol level and lysozyme activity were detected in HM and Co diet groups, and alternative complement pathway activity was detected in the HM diet group. However, significantly lower serum alanine aminotransferase and aspartate aminotransferase activities were obtained in all herbal diet groups compared with the control diet group. Pathogen challenge test by intraperitoneal injection of Vibrio anguillarum indicated that highest survival was obtained in the HM diet group followed by Ac, Co, Cf, and Mm diet groups. The lowest survival was obtained in the control group. These results reveal that medicinal herbs in diets enhance growth and some non-specific immunity of red sea bream.     

Periodic changes in the growth performance and biochemical composition of juvenile red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis> fed non-heated and heated squid and krill meal-based diets

In finfish aquaculture, fish meal is heated during the manufacturing process, which affects the digestibility and protein absorption by fish. However, manufactured fishmeal that is not heated does not undergo thermal denaturation. Few studies have investigated the effects of non-heated animal protein sources on the growth performance of fish. We investigated the effects of heated and non-heated squid and krill meal as diets for red sea bream. Five test diets were formulated to contain heated squid meal, non-heated squid meal, heated krill meal, non-heated krill meal, and fish meal as a control. Fifty fish (initial mean weight?=?3.5 g) were distributed in ten 100-l experimental tanks. Fish were fed one of the five diets 3 times daily until satiation for 5 weeks. Regarding growth performance, fish fed the krill meal diet exhibited better growth than those fed squid meal during the first week of the rearing period. However, the squid meal diet group showed better performance than the krill meal diet group during the third week. Moreover, differences in body weight among treatments were greater during the fifth week. Better weight gain and thermal growth coefficient were recorded in the non-heated diet groups than in the heated diet groups. Higher feed intake was observed in the non-heated diet groups than in the heated diet groups. These results suggest higher performance of non-heated squid and krill meal as the protein source of the red sea bream diet. Further, the suitability of the diet type (e.g., squid and krill) might depend on the feeding period and/or developmental stage of fish.     

Seagrass habitat reduces vulnerability of red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis> juveniles to piscivorous fish predator

ABSTRACT:   Predation experiments were conducted in mesocosms to test the hypothesis that habitat complexity affects vulnerability of red sea bream Pagrus major juveniles to piscivorous fish predators. Juvenile behavior was video-recorded for 6 h in two structurally different habitats: vegetated with sea grass Zostera marina , and unvegetated 0.5-t tanks. Association behavior with seagrass was observed in the vegetated tank throughout the trials. Predation experiments were conducted with 30 red sea bream juveniles (29.9 mm body length) exposed to two individuals of the piscivorous fish predator Chinese sea bass Lateolabrax sp. (261.6 mm), for 6 h in 1.0-t tanks. Predation rate (no. of fish predated on per predator per h) was significantly lower in the vegetated tank (0.02/predator per h) than in the unvegetated tank (0.27/predator per h). Present experiments indicate that habitat complexity reduces vulnerability of juvenile red sea bream to predation by piscivorous fish by serving as physical and/or visual barriers and limiting the predator's ability to pursue and capture prey. Seagrass beds in the shallow coastal waters around Japan are suggested to be an important nursery for red sea bream since they provide the juveniles with habitat complexity as well as serve as a feeding ground.     

Comparison of apparent digestibility coefficient among replicates and different stocking density in red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

Two consecutive feeding trials were conducted to know: (i) how the apparent digestibility coefficient (ADC) varies among replicates on different days and during the course of the experiment (Trial 1); and (ii) the effect of stocking density (e.g. 7.17, 5.56, and 3.92 kg fish/t water) on the ADC of nutrients and energy in red sea bream (Trial 2). In both trials, 0.5% Cr₂O₃ was used as an inert marker. In Trial 1, there were no significant differences in ADC within the replicates although the values on different days varied significantly during the second week. The ADC of all parameters in the third and fourth weeks was significantly higher than in the second week (P<0.05). Although statistically insignificant, the lower stocking densities (3.92 and 5.56 kg fish/t water) showed better growth performance and superior ADC of protein, lipid, and energy than the higher stocking density in Trial 2. The results indicated that it may be better to acclimate fish to the diet for at least three weeks to obtain more reliable data by using a single tank for each treatment for digestibility experiments. Results also indicated that growth and digestibility performance can be stimulated by controlling stocking density.     

Immune response of sea bass <Emphasis Type="Italic">Dicentrarchus labrax</Emphasis> to <Emphasis Type="Italic">Tenacibaculum maritimum</Emphasis> antigens

ABSTRACT:   Seawater fishes are affected by a pathology commonly called 'myxobacteriosis', caused by Tenacibaculum maritimum (formerly Flexibacter maritimus ). The disease is characterized by fin erosion and necrotic ulcers of skin and muscle, and by low but constant mortality in cultured marine fish; in Italy is one of the most important and widely spread diseases affecting sea bass Dicentrarchus labrax , gilthead seabream Sparus aurata , sharp-snouted bream Diplodus puntazzo , white bream Diplodus sargus , and six-tooted bream Dentex dentex . In order to obtain an effective vaccine against the disease, formalin killed cells (FKC), extracellular products (ECPs) and crude lipopolysaccharide (LPS) preparations were obtained from the T. maritimum strain SPVId and injected intraperitoneally twice into the sea bass. The fish immune response to the preparations was studied: agglutinating antibody titer and in vitro phagocytosis were determined after the first and second injection in order to evaluate whether the preparations are immunogenic or not and if the booster effect took place. The results show that FKC and LPS preparations increased the antibody titer after the first injection when compared to the control sea bass. Moreover, all the preparations stimulated a secondary (booster) response. In vitro phagocytosis of the total blood was significantly higher for all the preparations when compared to the controls, but the crude LPS immunized sea bass showed the highest activity.     

Spatial learning-induced <Emphasis Type="Italic">egr</Emphasis>-<Emphasis Type="Italic">1</Emphasis> expression in telencephalon of gold fish <Emphasis Type="Italic">Carassius auratus</Emphasis>

The immediate-early gene (egr-1) expression was used to examine the neuron’s response in telencephalon of goldfish during spatial learning in small space. Fishes were pre-exposed in the experimental apparatus and trained to pick food from the tray in a rectangular-shaped arena. The apparatus was divided into identical compartments comprising three gates to provide different spatial tasks. After the fish learned to pass through the gate one, two more gates were introduced one by one. Fish made more number of attempts and took longer time (P < 0.05) to pass through the first gate than the gate two or three. This active learning induces the expression of egr-1 in telencephalon as established by western blot analysis. Subsequently, the fish learn quickly to cross the similar type of second and third gate and make fewer errors with a corresponding decline in the level of egr-1 expression. As the fish learned to pass through all the three gates, third gate was replaced by modified gate three. Interestingly, the level of egr-1 expression increased again, when the fish exhibit a high exploratory behavior to cross the modified gate three. The present study shows that egr-1 expression is induced in the telencephalon of goldfish while intensively acquiring geometric spatial information to pass through the gates.