Reproduction,growth and stress response in adult red sea bream,Pagrus major (Temminck & Schlegel) exposed to different photoperiods at spawning season

Adult red sea bream, Pagrus major (body weight, 1.0–2.0 kg) was exposed to three photoperiods [12 h light:12 h dark (12L:12D), 16 h light:8 h dark (16L:8D) and 24 h light:0 h dark (24L:0D)] from 2 months before spawning till the end of the spawning season to investigate growth, spawning and stress response. During the spawning season, tanks were checked every morning for spawned eggs. The growth performance in fish under 24L:0D was stimulated with significantly higher feed intake than those under other photoperiods (P<0.05). The number of eggs and gonadal histology confirmed that three and five females out of six in each of duplicate tanks of the 16L:8D treatment spawned. In contrast, only two out of six females in one tank of the 24L:0D treatment spawned, and no spawns were observed in the 12L:12D treatment. At the end of the spawning period, both 17β‐estradiol and testosterone levels were significantly higher in fish exposed to 16L:8D followed by 12L:12D and 24L:0D photoperiods (P<0.05). Photoperiod manipulation did not cause significant stress response in fish (P>0.05). The results suggest that stimulating the growth performance of red sea bream at reproductive stage with a 24L:0D photoperiod is possible if the fish are subjected to this photoperiod long before the onset of the spawning season.     

Hypoxic conditions induce centrum defects in red sea bream Pagrus major (Temminck and Schlegel)

A previous study elucidated that an extreme hypoxia during somitogenesis induced the most frequent skeletal malformation centrum defects in red sea bream (RSB), Pagrus major. In this study, details of the hypoxic conditions to induce them in RSB, dissolved oxygen (DO) concentration and exposure time to hypoxia, were investigated. Fertilized eggs were exposed to seawater of six DO concentrations (0%, 10%, 25%, 50%, 75% and 100% of saturation) for seven different periods (5, 10, 30, 60, 120, 240 and 360 min) during somitogenesis. Somitic disturbances in newly hatched larvae were induced by exposure to 0% and 10% DO concentration for 10 and 120 min and longer respectively. Rearing eggs exposed to hypoxic condition of 10% DO for 240 min for 40 days post‐hatch showed that the location and the frequency of somitic disturbances in larvae and centrum defects in juveniles were significantly correlated (P<0.01). Dissolved oxygen concentration of the interstitial water in the egg high density layer formed at the water surface in a stationary state abruptly decreased to 3.7% within 7 min. Centrum defect induction by exposure of eggs to extreme low DO concentrations for a short period, which is the probable situation in the practical juvenile production, suggests that careful maintenance of DO concentration is important in the incubating water of fertilized eggs during egg sorting and transportation, where eggs are made into a pile and undergo hypoxia, for the prevention of centrum defects.     

Ultrasonic immunization of sea bream, Pagrus major (Temminck & Schlegel), with a mixed vaccine against Vibrio alginolyticus and V. anguillarum

In order to clarify the effectiveness of ultrasonication on vaccine delivery, juvenile sea bream, Pagrus major , were treated with eight different ultrasonic methods. A mixed vaccine against Vibrio alginolyticus and V. anguillarum was used to immunize the fish . The intensity and frequency of the ultrasound were 280 mW cm^–2 and 35 kHz, respectively. The ultrasonic methods included continuous or pulsed ultrasound for 3 min, and continuous or pulsed ultrasound for 3 min before and/or after immersion for 3 min. Of all the eight ultrasonic methods tested, `pulsed ultrasound followed by immersion' and `immersion, pulsed ultrasound, and followed by immersion again' provided the best protection, which were comparable with protection of fish immunized by intraperitoneal injection. Moreover, the convenience of applying these two ultrasonic methods for immunization was comparable with the immersion method and was much better than intraperitoneal injection. If 2 × 10⁸ CFU mL^–1 of this mixed vaccine was used for vaccination repeatedly five times by ultrasonic methods, it could still produce good protection for the immunized sea bream. Therefore, the ultrasonic method is an effective and practical approach for fish vaccination on a large scale.     

Oxygen deficiency during somitogenesis causes centrum defects in red sea bream, Pagrus major (Temminck et Schlegel)

Vertebral deformities in red sea bream, Pagrus major, remain serious obstacles to the improvement of seedling quality for its aquaculture. However, the causalities of the deformities remain unclear and prevention methods have not yet been established. In this paper, oxygen deficiency during somitogenesis was demonstrated to cause centrum defects (formerly called fused vertebrae in many cases), which are the major vertebral deformity in cultured red sea bream. An induction experiment of centrum defects was conducted by placing fertilized red sea bream eggs under low dissolved oxygen conditions (10.3–16.6%). The low oxygen treatment was carried out for five different developmental stages of embryo: two‐cell stage to blastula stage; gastrula stage; three to 10 somites stage; 11–17 somites stage and 18–24 somites stage. Oxygen deficiency during somitogenesis induced a high incidence of centrum defects. In contrast, it hardly induced centrum defects during the other stages. The dissolved oxygen concentration in the rearing water should be carefully regulated for fertilized eggs, especially during somitogenesis to reduce the incidence of vertebral deformities in the red sea bream.     

Henneguya pagri n. sp. (Myxozoa: Myxosporea) causing cardiac henneguyosis in red sea bream, Pagrus major (Temminck & Schlegel)

Henneguya pagri n. sp. (Myxozoa: Myxosporea) is described from netpen-cultured red sea bream, Pagrus major, in Japan. Affected fish displayed anaemic gills, an enlarged bulbus arteriosus and internal haemorrhaging in the pericardial cavity. Disease outbreaks occurred during the summer and the prevalence of infection reached 75% in August. Plasmodia of H. pagri developed in the bulbus arteriosus of red sea bream, in which degenerative cardiomyopathy was observed. Following maturation of the parasite, the influx of spores into the gills caused congestion of the gill capillaries, resulting in proliferative branchitis. Spores (10.5 x 7.5 microm) were ovoid with two caudal appendages (29.6 microm in average length), with a whip-like extension from the end. Partial small subunit ribosomal RNA gene sequences of H. pagri are closely related to those of H. lateolabracis infecting the bulbus arteriosus of Chinese sea bass, Lateolabrax sp.     

Effect of rotifer enrichment with herbal extracts on growth and resistance of red sea bream,Pagrus major (Temminck & Schlegel) larvae against Vibrio anguillarum

The effects of the dietary medicinal herbs Massa medicata, Crataegi fructus (Cf), Artemisia capillaries, Cnidium officinale and their mixture (HM), on growth and resistance against Vibrio anguillarum were examined using larval red sea bream, Pagrus major. The methanol extracts of Cf and HM inhibited the proliferation of seven infectious bacterial strains including the genera Aeromonas, Edwasiella, Photobavterium, Pseudomonas and Vibrio. Rotifers enriched with emulsified bonito oil containing extracts of Cf and HM lowered the bacterial counts in their body on TCBS medium for the genus Vibrio in comparison with rotifers enriched by emulsified bonito oil alone. Although statistically not significant, larvae fed rotifer fortified with Cf from 3 to 20 days after hatching showed the best growth and final body weight followed by those given rotifer with HM and control enrichment. However, final body length of larvae fed rotifer with Cf was significantly larger than larvae fed on rotifers enriched with other herb extracts. After a challenge test with V. anguillarum, survival of larvae fed rotifer with Cf and HM was significantly higher than for the control treatment. These results revealed that Cf and HM in rotifer is useful to promote growth and resistance against V. anguillarum in red sea bream larvae, providing a new technology for mass production of disease‐resistant fry and fingerlings.     

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.     

Establishment and characterization of a fibroblast cell line derived from the dorsal fin of red sea bream,Pagrus major (Temminck & Schlegel)

The establishment and partial characterization of a continuous cell line from the dorsal fin of red sea bream, Pagrus major, are described. The cell line, designated RSBF‐2, has been subcultured for more than 100 passages since its initiation in November 2000. It was optimally maintained at 28 °C in Leibovitz L‐15 medium with 10% foetal bovine serum. Propagation of RSBF‐2 cells was serum dependent and exhibited low plating efficiency (<1.7%). Aside from long‐term cryopreservation, the cells could also be kept at 4 °C for 72 days. The distribution of the chromosome number was 38–98 with a mode of 48. The RSBF‐2 cell line was susceptible to red sea bream iridovirus but only produced a few rounded and refractory cells. Virus‐inoculated RSBF‐2 cells were then subcultured to generate a persistently infected cell line. RSBF‐2 was also very sensitive to the extracellular products of Photobacterium damselae ssp. piscicida and produced significant fluorescent signals after transfection with pEGFP‐C3. Analysis of mitochondrial cytochrome b gene sequences revealed 99% identity between the cell line and Pagrus major.     

Optimum dietary taurine level in casein-based diet for juvenile red sea bream Pagrus major

ABSTRACT:   This study was conducted to investigate the effect of dietary taurine and cholyltaurine (C-tau) on growth and body composition of juvenile red sea bream Pagrus major . Semi-purified casein-based diets supplemented with 0 (control diet), 0.1, 0.3, 0.5 and 0.7% taurine and 0.5% C-tau were fed to red sea bream (average body weight 4.7 g) for 6 weeks at 20°C. The growth and feed efficiency were the lowest in fish fed the control diet. Taurine supplementation improved the growth and feed efficiency of fish dose-dependently, and the taurine requirement was estimated as 0.52% in terms of optimizing growth and 0.48% in terms of optimizing feed efficiency. Taurine content in the whole body and liver increased with the dietary taurine level. Supplemental C-tau at the 0.5% level had limited effects on the growth and no effect on body taurine, biliary bile salt and liver fat contents. From these results it can be inferred that the optimal dietary taurine requirement of juvenile red sea bream is 0.5% on a dry weight basis, and that the supplementation of taurine in the diet not only improves the growth but also increases hepatic lipid levels of red sea bream juveniles.     

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.     

Inosine supplementation effectively provokes the growth,immune response,oxidative stress resistance and intestinal morphology of juvenile red sea bream,Pagrus major

A feeding trial was conducted to determine effects of dietary inosine on growth, immune responses, oxidative stress resistance and intestinal morphology of juvenile red sea bream. A semi‐purified basal diet supplemented with 0 (D1, control), 2 g kg^?1 (D2), 4 g kg^?1 (D3), 6 g kg^?1 (D4) and 8 g kg^?1 (D5) dietary inosine, respectively, to formulate five experimental diets. Each diet was randomly allocated to triplicate groups of fish (initial weight: 8 g). After 50 days of feeding trial, fish fed diets with 4 g kg^?1 inosine had the highest (p < .05) final weight, weight gain and specific growth rate. Similarly, improved feed intake, feed conversion ratio and protein efficiency ratio were also found at 4 g kg^?1 inosine supplemented group. Some non‐specific immune parameters such as total serum protein, lysozyme activity and bactericidal activity tended to be higher for fish fed diets supplemented with inosine. Peroxidase and catalase activity also influenced (p < .05) by dietary inosine supplementation. Fish fed 2 g kg^?1 and 4 g kg^?1 inosine supplemented diets showed the least oxidative stress condition. Inosine supplementation significantly increased (p < .05) anterior enterocyte height and posterior fold height, enterocyte height and microvillus height compared with control. We concluded that 4 g kg^?1 dietary inosine supplementation effectively provokes growth and health performance of red sea bream by increasing growth, immune response, oxidative stress resistance and intestinal health condition.     

Secondary nursing of the red sea bream,Chrysophrys major (Temminck & Schlegel), in an earth pond

Fry of the red sea bream, Chrysophrys major (Temminck & Schlegel), were nursed in an earthern pond for 40 days. Samples of 50 fry were weighed and measured individually at 10-day intervals. Growth of fry was faster in this pond than in the open sea, presumably as a result of the higher temperature in the pond. The high mortality during the first and the second 10-day periods was probably caused by handling and incorrect feeding. The frequency distribution of weight was skewed to the right for all samples, with the highest coefficient of skewness at the first and last weighings.     

Diluted rotifer, Brachionus plicatilis (Muller), extract as culture medium stimulates hatching of red sea bream, Pagrus major (Temminck & Schlegel)

Hatching rates and hatching times (50% hatch) of red sea bream, Pagrus major (Temminck & Schlegel), eggs in culture media that were prepared to dilute rotifer Brachionus plicatilis Muller extract at 4000-, 6000-. 8000- and 10 000-fold with artificial sea water (σ₁₅, = 24.0) were investigated. Although differences in hatching rate between the culture media and control artificial sea water were negligible, hatching time was significantly shorter in media at 4000- and 6000-fold dilutions than in media at 8000- and 10 000-fold dilutions and in the control artificial sea water, when the eggs immediately post spawning were incubated. The hatching rates of eggs at three developmental stages, immediately post-spawning, gastrula stage and heart-beat initiation stage, incubated respectively in the medium at 6000-fold dilution and the control, did not differ either, yet the hatching times at each egg stage in the medium were significantly shorter than that in the control. These results suggest that hatching enzyme secretion from the hatching glands is promoted to perceive chemical stimuli in the water-soluble fraction of rotifer extract via the embryo's chemoreceptors, from the heart-beat initiation stage to immediately before hatching, thus shortening the hatching time.     

真鲷精子诱导牙鲆减数分裂雌核发育

采用真鲷精子激活牙鲆卵子,经(0±0.5)℃冷休克处理,诱导了牙鲆减数分裂雌核发育。灭活真鲷精子,紫外线照射剂量3.4mJ/cm2时孵化率最低;随着照射剂量的增加,孵化率恢复,照射剂量73mJ/cm2时,孵化率最高,呈现典型的Hertwig效应。用流式细胞仪检测倍性,未经冷休克处理的胚胎全部为单倍体,经冷休克处理的均为二倍体,表明精子灭活有效。冷休克实验结果表明,冷休克起始时间2～5min均有效,以3min为最好;冷休克持续时间30～90min均有效,以45min为最好。综合两项因素,授精3min后,休克时间持续45min组的授精率和孵化率均为最高,与其他处理组差异显著(P<0.05)。RAPD分析结果显示,异源精子的遗传信息未在雌核发育二倍体的电泳图中出现,表明其遗传信息没有传递给子代。     

Availability of black soldier fly meal as an alternative protein source to fish meal in red sea bream (Pagrus major,Temminck & Schlegel) fingerling diets

Black soldier fly meal (BM) is an outstanding candidate as a fish meal (FM) substitute because it contains relatively high protein and essential amino acids. In this study, we replaced FM in red sea bream diets (Pagrus major) with BM to investigate its effect on growth and feed utilization. Six isonitrogenous and isolipidic experimental diets were prepared by substituting 0%, 20%, 40%, 60%, 80% and 100% FM protein with BM (Control, BM20, BM40, BM60, BM80 and BM100, respectively). After the 8-week feeding trial, final body weight, weight gain, specific growth rate and feed efficiency decreased linearly with increasing dietary BM level (p < 0.05). Apparent digestibility of protein and fat and serum total cholesterol concentration decreased with increasing dietary BM levels (p < 0.05) Final body weight, WG, SGR and FE against dietary BM level had breaking points of 76.2%, 41.7%, 76.5% and 60.0%, respectively, in segmental regression analysis. In conclusion, the results suggest that BM can replace a maximum of 41.7% of FM in the diet of red sea bream without compromising growth performance or feed efficiency for 56 days.     

Genetic diversity of red sea bream Pagrus major in western Japan in relation to stock enhancement

SUMMARY: Stock enhancement is used in Japan as a tool to help the replenishment of wild populations of red sea bream Pagrus major . In this study, we analyzed the genetic diversity and composition of wild red sea bream at seven locations around Shikoku Island, South-west Japan, using three microsatellite loci. This analysis was done to test the hypothesis that: (i) red sea bream comprises a single Mendelian population along Japan; and (ii) stock enhancement programs around Shikoku Island are causing genetic differentiation among wild stocks. The results indicated that some locations from the Shikoku area were not significantly different from the rest of Japan, supporting the hypothesis of a single Mendelian population. Significant departures from Hardy–Weinberg equilibrium and significant pairwise F_ST among locations indicated genetic instability within this region. We suggest that the stock enhancement programs made in the region are the possible cause of this genetic instability. A management scheme for the hatcheries involved in the stock enhancement of red sea bream is presented.     

日本鹿儿岛湾真鲷的资源评估和增殖策略

通过世代分析对1990年至2002年鹿儿岛湾真鲷（Pagrus major）自然种群和放流种群的种群大小进行了评估,评估模型考虑了渔业资源管理和增殖放流措施。结果表明放流种群的规模有所减少,而自然种群的规模维持不变。真鲷自然种群的捕获量日趋减少,而放流种群的捕获量已超过50吨。研究结果表明增殖700000—800000尾真鲷种苗与其50吨的捕捞量相协调。在各种捕获量和放流量情况下,采用种群动力学模型预测了2017年真鲷的捕获量,其结果表明提高放流真鲷的数量能增加其捕获量。     

Effect of dietary lipid oxidation with vitamin C and E supplementation on fillet quality of red sea bream,Pagrus major (Temminck & Schlegel) during storage

An experiment was conducted to determine the effects of different levels of dietary vitamin C (VC) and E (VE) supplementation on fillet quality of red sea bream fed oxidized fish oil (OFO). Fish with an average body weight of 205.0 g were fed four test diets for 9 weeks. Control diet contained fresh fish oil (FFO) with 100 mg kg^?1 of VE and 500 mg kg^?1 of VC (FFO100E/500C). The other three diets contained OFO with varying levels of VE (mg kg^?1) and VC (mg kg^?1) (OFO100E/500C, OFO200E/500C and OFO200E/1000C). After feeding trial, two fillets from each fish by hand filleting were stored in a refrigerator at 4°C for 96 h during analyses. Results showed that fish fed OFO increased fillet thiobarbituric acid reactive substances (TBARS) and K‐value, and decreased fillet VC and VE concentrations during storage time. Supplementation of VC did not have any detectable effect on fillet quality. Increasing dietary VE supplementation increased fillet VE concentrations, reduced fillet TBARS and K‐value values of red sea bream. Therefore, we suggest that dietary supplementation of 200 mg kg^?1 of vitamin E could improve fillet oxidative stability of red sea bream fed OFO.     

Effects of dietary copper nanoparticles and vitamin C supplementations on growth performance,immune response and stress resistance of red sea bream,Pagrus major

A 60‐day feeding trial was conducted to determine the effects of copper nanoparticles (Cu‐NPs) and vitamin C (VC) on red sea bream. Besides the control diet (D1), six diets were supplemented with Cu‐NPs and VC [0/800 (D2), 0/1,000 (D3), 0/1,200 (D4), 2/800 (D5), 2/1,000 (D6) and 2/1,200 (D7) mg Cu‐NPs/VC per kg]. Cu‐NP was a significant factor on final weight (FBW), weight gain (WG) and specific growth rate (SGR), feed intake (FI), feed (FER) and protein efficiency ratios (PER), protein gain (PG) and protein retention (PR), body protein and lipid contents, protease (PA) and bactericidal activities (BA) and tolerance against stress (LT50%) (p < .05). In addition, BA and LT50% were significantly affected by either Cu‐NPs or VC (p < .05). Fish fed Cu‐NPs or/and VC‐supplemented diets showed higher FBW, WG, SGR, PG, PR, FI, PA, LA and BA values when compared with the control group (p < .05). FER, PER and body lipid content were significantly enhanced in D4, D5, D6 and D7 groups; meanwhile, body protein and LT50% were significantly enhanced in D5, D6 and D7 groups when compared with D1 group (p < .05). In conclusion, dietary Cu‐NPs or/and VC improved growth and health of red sea bream.     

Photoperiod influences the growth, food intake, feed efficiency and digestibility of red sea bream (Pagrus major)

Two consecutive trials were conducted to investigate the effects of photoperiod manipulation on growth rate, food intake and feed conversion efficiency (Trial 1), and the digestibility of nutrients and energy (Trial 2) in red sea bream, Pagrus major (body weight 19–120 g). Fish were exposed to four photoperiods (6L:6D, 12L:12D, 16L:8D and 24L:0D) with light intensity 1500 lx on the water surface. The fish were fed with a commercial diet to apparent satiation. In Trial 2, 0.5% chromic oxide (Cr₂O₃) was used as an inert marker. Significantly higher weight gain and specific growth rates were observed in fish exposed to a 24L:0D photoperiod followed by 16L:8D, 6L:6D and 12L:12D photoperiods (P < 0.05). Food intake and feed conversion efficiency (FCE) were also significantly higher in fish exposed to 24L:0D followed by 16L:8D, 6L:6D and 12L:12D photoperiods (P < 0.05). Fish exposed to 24L:0D and 16L:8D photoperiods showed significantly higher lipid and energy digestibility than those exposed to a 12L:12D photoperiod (P < 0.05). The results demonstrated that the enhancement of growth performances under 16L:8D and 24L:0D photoperiods were attributed to improved appetite, greater food intake and higher feed conversion efficiency as well as higher digestibility.