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.     

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.     

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.     

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.     

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.     

Microsatellite multiplex panels for population genetic analysis of red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

In this study, multiplex PCR panels were developed to amplify 20 microsatellite markers for red sea bream, Pagrus major, with the aim of reducing labor and material costs associated with genetic analysis of this species. The usefulness of these panels was validated in 200 fish collected at five sampling locations. The occurrence of null alleles was suggested at five markers, which were dropped from further analysis. The remaining 15 microsatellite loci showed high levels of heterozygosity (H _E range 0.34–0.97, H _O range 0.32–1) and a wide range in the number of alleles per locus (A; range 5–46). Increasing the number of microsatellite loci from three to ten and 15 improved the performance of the panels for population differentiation (F _ST) and estimation of Nei’s (D _S) genetic distance. The results of this study confirm the usefulness of genotyping a large number of microsatellite DNA markers to expand our knowledge of red sea bream population genetics.     

Effects of fish size and water temperature on the acute toxicity of boron to Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> and red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

ABSTRACT:     The acute toxicities of boron were examined for Japanese flounder Paralichthys olivaceus and red sea bream Pagrus major in terms of fish size and water temperature. Japanese flounder of 0.1–70.0 g and red sea bream of 0.6–20.3 g were exposed to different concentrations of boron for 96 h at 20°C under semi-static conditions. In both fish species, the median lethal concentration (LC₅₀) for 96 h of boron increased linearly with increasing fish size, ranging from 108 to 252 mg B/L for the flounder, and from 97 to 172 mg B/L for the sea bream. The effect of water temperature on acute toxicity was examined for Japanese flounder of 0.6 and 1.5 g at 10, 15, 20 and 25°C, and for red sea bream of 0.6 and 2.4 g at 12, 15, 20 and 25°C. The toxicity of boron for the flounder increased linearly with increasing water temperature. The 96 h LC₅₀ values ranged from 299 to 108 mg B/L for the 0.6 g flounder and from 350 to 113 mg B/L for the 1.5 g flounder. A similar trend was shown for the 2.4 g red sea bream; however, the relationship for the 0.6 g red sea bream was not significant.     

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.     

Supplemental effect of <Emphasis Type="Italic">Porphyra</Emphasis> spheroplasts on the growth and feed utilization of black sea bream

ABSTRACT:   A 56-day feeding trial was conducted to evaluate the effects of Porphyra spheroplasts (PS) as a feed additive on growth, carcass composition and feed utilization of black sea bream. Four experimental diets with or without PS (0, 1, 3, 5% inclusion level) were formulated to determine the proper incorporation level for best growth and utilization using white fishmeal as the principal protein source. Three replicate groups of fish averaging 1.28 ± 0.05 g (mean ± SD) were maintained in flow-through thermo-controlled sea water (32–34 psu, 25°C) aquaria (100 L) under the laboratory conditions (L12 : D12) and fed three times a day at 09:00, 13:00 and 17:00 hours by hand at approximately 3% body weight on each diet. The rearing trial revealed that growth performance (in terms of weight gain, specific growth rate) are almost the same level in all the dietary groups, nevertheless the survival, nutrient utilizations and retentions (feed efficiency, protein efficiency ratio, protein retention rate, lipid retention rate) were significantly higher ( P  < 0.05) in a group fed on the diet containing 3% spheroplasts. Further, the fish fed on the diet containing PS had significantly higher ( P  < 0.05) crude lipid level in their dorsal muscles and viscera. The fish fed a PS-based diet showed higher level of eicosapentaenoic acid and docosahexaenoic acid (DHA) in viscera and DHA in dorsal muscles. The results obtained in this feeding trial suggest that the optimum dietary PS supplementation level up to 3% for black sea bream can be considered as improved consequence on growth performance, nutrient utilization or body composition were noticed.     

Reproductive cycle of yellow sea bream <Emphasis Type="Italic">Dentex tumifrons</Emphasis> in Wakasa Bay,the Sea of Japan off central Honshu

ABSTRACT:   To determine the spawning season and reproductive cycle of the yellow sea bream, Dentex tumifrons , in Wakasa Bay, in the Sea of Japan off central Honshu, seasonal change in the gonadosomatic index ( GSI ) was examined and histological observations of both ovaries and testes were conducted. The female GSI gradually increased from May, maintained a high level between August and October and then rapidly decreased. The histological observations of the ovary showed that vitellogenesis began from May and developed until September. Spent fish began to be observed in October and all the fish collected in December were spent. There was only one peak in the seasonal change in GSI of males and females in Wakasa Bay. Thus, yellow sea bream spawn only in autumn in Wakasa Bay. Differences in the reproductive cycle and number of spawning seasons were found between the two different latitudinal areas, Wakasa Bay and the East China Sea (low latitudinal area) where the spawning seasons were spring and autumn. An increase in water temperature is an important factor to initiate gonadal maturation (recrudescence) and is terminated by the short day-length in Wakasa Bay.     

Vertebral deformities in cultured red sea bream, Pagrus major, Temminck and Schlegel

Vertebral deformities were investigated in cultured red sea bream, Pagrus major. In the field, deformities in seedlings were categorized and their incidence was calculated. In the laboratory, the symptoms of major vertebral deformities were examined morphologically using radiographs and by making transparent skeletal specimens. The internal structure of deformed vertebrae was examined histologically. The shortened body condition had the highest incidence (0.9–8.3%) of all deformities in the seedlings. In individuals with the shortened body condition, the ratio of trunk and caudal part length to body height was smaller. These fish had skeletal anomalies in the vertebrae, mainly centrum defects (64.3%) or undersized centrums (25.2%). The specimens with centrum defects had a characteristic anomaly in the vertebrae, with plural pairs of neural and haemal spines on a single centrum. This anomaly was frequently observed in the posterior abdominal vertebrae. The internal skeletal structure of such abnormal centrums was basically the same as that of normal centrums. In all the specimens with undersized centrums, both the centrum length and diameter were shorter than normal except for the first and second centrum, and urostyle.     

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.     

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.     

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.     

Changes in muscle collagen content during <Emphasis Type="Italic">post mortem</Emphasis> storage of farmed sea bream (<Emphasis Type="Italic">Sparus aurata</Emphasis>): influence on textural properties

The changes in collagen content and its solubility in sea bream muscle were studied for variable storage times following the death of the fish, and these variables were related to the evolution of physical parameters important for consumer acceptance: firmness and water-holding capacity (WHC). The results show that the collagen content in muscle diminished slightly over storage time and that this variable was directly related to firmness but inversely related to the water-holding capacity. With regard to collagen solubility, a decline was detected in acid-soluble collagen (ASC) in the first few post mortem hours, perhaps related to the end of rigor mortis that occurs at these stages. Pepsin-soluble collagen (PSC) increased, while insoluble collagen (ISC) decreased from 96 h, coinciding with a loss of firmness. This softening can be explained as a result of specific collagenases acting on the insoluble fraction of the collagen.     

Predation on fish larvae by moon jellyfish <Emphasis Type="Italic">Aurelia aurita</Emphasis> under low dissolved oxygen concentrations

ABSTRACT:   Laboratory experiments were conducted to test the hypothesis that low dissolved oxygen concentrations have the potential to enhance the predation rate on fish larvae by moon jellyfish Aurelia aurita which is increasing in abundance in the coastal waters of Japan. Larvae of the red sea bream Pagrus major in four size classes (2.9, 4.1, 6.2 and 8.6 mm in standard length) were used as prey in a short-term predation experiment. No change in the bell contraction rate of the jellyfish (mean bell diameter, 100.4 ± 10.2 mm) was observed at the oxygen concentrations tested (1, 2 and 4 mg/L, and air-saturated, 5.5–6.0 mg/L), suggesting a strong tolerance to a decline in the oxygen concentration. More than 80% of the 2.5 and 4.1-mm size-class larvae were predated on by the jellyfish at all oxygen concentrations during the 15-min trials in 10-L tanks. The 6.2 and 8.6-mm size-class larvae were able to escape from the jellyfish apparently due to their developed swimming ability at the two higher dissolved oxygen concentrations. There was an increase in predation on larvae of these two size classes at the two lower dissolved oxygen concentrations. Predation by moon jellyfish on fish larvae can be more intense at low dissolved oxygen concentrations that commonly occur in the coastal waters of Japan.     

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.     

Studies on epizootic iridovirus infection among red sea bream,Pagrus major (Temminck & Schlegel), cultured in Taiwan

Since 1993, an epizootic viral disease has occurred in net-cage cultured red sea bream, Pagrus major (Temminck & Schlegel), in Peng-hu Island located on the south-western coast of Taiwan. The diseased fish exhibited abnormal swimming and were lethargic, but few visible external signs were observed. The cumulative mortality because of the disease sometimes reached 50-90% over 2 months. Histopathogical studies of the affected fish showed enlarged basophilic cells in the gill, kidney, heart, liver and spleen. These necrotic cells were Feulgen-positive and stained blue using Giemsa. Transmission electron microscopy revealed icosahedral virions in the cytoplasm of the necrotic cells. The viral particles consisted of a central nucleocapsid (75-80 nm) and envelope, and were 120-150 nm in diameter. These results suggest that the virus belongs to the Iridoviridae. Using polymerase chain reaction (PCR), approximately 570 bp fragments were produced from the viral DNA using as a template 1-F and 1-R primers derived from red seabream iridovirus (RSIV) from red sea bream in Japan. Similar results were also obtained using nested-PCR with different primer sets (1-F, 2-R and 2-F, 1-R). Although the size and some features of epizootics of this virus differed from RSIV in Japan, it shows close genetic affinities with the latter and it is suggested that RSIV has been introduced to Taiwan.