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.     

Ontogeny of tolerance to and avoidance of ultraviolet radiation in red sea bream Pagrus major and black sea bream Acanthopagrus schlegeli

ABSTRACT:   Ontogenetic changes of tolerance to, and avoidance of, ultraviolet-B radiation (UV-B) were examined in red sea bream Pagrus major and black sea bream Acanthopagrus schlegeli . In the tolerance experiment, larvae and juveniles (age 13–46 days) were put in beakers, and were exposed to one of five different levels of UV-B radiation (1.8, 1.1, 0.2, 0.1, and 0 W/m²) for one hour. Their survival rates were calculated either 12 or 24 h later. In the avoidance experiment, fish (age 3–49 days) were put in a long experimental tank, half of which was covered with UV-blocking film and placed under two levels of UV-B radiation (1.1 and 0.2 W/m²), and their avoidance indices were calculated. Black sea bream had significantly better survival compared to red sea bream for most ages. Only black sea bream of ages 37 and 49 days showed significant avoidance of UV radiation under the higher level of UV-B, whereas both species did not show avoidance on any days at the lower level. The present results suggest that black sea bream are significantly better adapted to habitats with high UV-B radiation, than red sea bream, reflecting that back sea bream live in shallower waters through their early life stages.     

Effect of dietary taurine enhancement on growth and development in red sea bream Pagrus major larvae

This study investigated the effect of feedings taurine‐enriched rotifers on the growth and development of larval red sea bream (RSB). Rotifers incubated in taurine‐enriched water at a taurine concentration of 800 mg L^?1 (T‐800) and 0 mg L^?1 (T‐0) were fed to larvae from 3 to 20 days after hatching (DAH). Notochord length, body weight and specific growth rate of T‐800 group were significantly greater than those of T‐0 at 14, 17, 9–11 and 18–20 DAH. Taurine content of larvae in the T‐800 group increased rapidly from 11 DAH and thereafter remained significantly higher than T‐0. Flexion larvae firstly appeared in both groups at 8 DAH, however, at 20 DAH post‐flexion larvae were significantly more abundant in T‐800 than T‐0. While nucleic acid and protein contents (μg mg^?1 wet fish) showed remarkable changes, ontogenetic growth in RSB larvae stage was observed to switch from hyperplastic growth to hypertrophic growth with the start of the flexion stage. Although a similar change in nucleic acid contents was observed between the two groups, the protein content (μg fish^?1) and protein/DNA ratio of T‐800 remained higher than that of T‐0 during the hypertrophic growth period. These results suggest that dietary taurine accelerates the growth and development in RSB larvae especially during hypertrophic growth (flexion stage) after the early hyperplastic growth.     

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.     

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.     

Morphological aspects of feeding and improvement in feeding ability in the early larval stages of red sea bream Pagrus major

The present study examined the appearance, ossification and growth of the bones that form the oral cavity in early larval stages of laboratory reared red sea bream ( Pagrus major ) for 380 h after hatching. The fundamental elements of the oral cavity appeared 11 h after initial mouth opening (HAMO). Development in the red sea bream, based on the osteological development of the feeding apparatus, was divided into three phases following the first feeding (24 HAMO; mean total body length 3.3 mm). The first phase was the early sucking phase (24 to 80–100 HAMO; approximately 3.9 mm), during which the head and bones increased in size. The intensified sucking phase (to 200–220 HAMO; approximately 4.9 mm) was defined by the appearance of new structural elements and a continued enlargement of the head and bones. Finally, during the transition phase (beyond 300 HAMO; approximately 5.6 mm), larvae used grasping as well as sucking to feed, new elements appeared, ossification began, size increased and teeth were acquired. As the larvae advanced through these three phases, the ability to feed by sucking was enhanced by the appearance and growth of new bones. The developmental phases appear to be linked to the transition from endogenous to exogenous nutrition resources under laboratory rearing conditions and to diversification in the size and components of wild food organisms.     

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.     

Fish meal replacement by soy protein from soymilk in the diets of red sea bream (Pagrus major)

Six isoenergetic diets were formulated as follows: fish meal (FM) 700 g kg^–1 (control, C), FM 300 g kg^–1 + soy protein concentrate 300 g kg^–1 (SPC), FM 300 g kg^–1 + enzyme‐treated SPC 300 g kg^–1 (ESC), FM 170 g kg^–1 + soy protein isolate 300 g kg^–1 (SPI), FM 160 g kg^–1 + enzyme‐treated SPI 300 g kg^–1 (ESI) and FM 150 g kg^–1 + conglycinin 300 g kg^–1(CG). Forty fish (3.9 g) were randomly distributed into each of eighteen 300‐L tanks, fed twice daily until satiation for 8 weeks. The final body weight, specific growth rate and condition factor did not show significant differences among the fish fed with diets C, SPC, ESC and ESI (p > .05). The survival was significantly lower in fish fed with diets SPI and CG. Feed efficiency was significantly higher in fish fed with diets SPC and C than in fish fed with other diets (p < .05). There were no significant differences in nutrients retention efficiencies in fish fed with diets C, SPC, ESC and ESI. A significantly higher phosphorus retention efficiency in fish fed with soymilk protein diets resulted in lower phosphorus discharge to the environment (p < .05). These results suggest that the soymilk proteins can comfortably replace 570–770 g FM kg^–1 diet of red sea bream juvenile, which will ensure significant ecological benefits through reducing phosphorus load to the environment.     

Teratogenic effects of isolated and combined short‐term hypercapnia and hypoxia on red sea bream (Pagrus major) embryos

Some vertebral anomalies in cultured fish arise from unusual environmental conditions during embryogenesis. We investigated the individual and combined teratogenic effects of short‐term hypercapnia and hypoxia on embryos of red sea bream (RSB), Pagrus major. Ten‐somite stage embryos were exposed to hypercapnia (60 and 120 mg/L dissolved carbon dioxide: DCD) and hypoxia (0%, 10%, 25%, 50%, and 75% dissolved oxygen: DO) independently and concomitantly for seven different periods (0, 30, 60, 90, 120, 150 and 180 min) to examine somitic disturbances at hatching, which are prodromal symptoms of centrum defects. Another experiment examined vertebral anomalies in juveniles raised from eggs exposed to hypercapnia (120 mg/L DCD) and hypoxia (10% DO) independently and concomitantly. Short‐time exposures (30–180 min) to hypercapnia (60 and 120 mg/L) and hypoxia (10% DO or less) independently and additively caused larval somitic disturbances and juvenile centrum defects. The results indicate that short‐term hypercapnia and hypoxia generally have the same teratogenic effect on embryos, although there were some differences in the locations of the somitic disturbances and centrum defects, with additive teratogenicity when the conditions were combined. These results suggest the necessity of maintaining appropriate DCD and DO concentrations during egg incubation and transport and during reproduction in recirculating aquaculture.     

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.     

Genetic diversity and structure of domesticated strains of red sea bream,Pagrus major,inferred from microsatellite DNA markers

The genetic diversity and structure of nine domesticated strains of red sea bream used in a private hatchery were studied and compared to a wild population. A total of 313 individuals were genotyped at eight microsatellite loci. Average number of alleles per locus ranged from 5.5 to 9.4 in domesticated strains, but that of the wild population was 28.4. Heterozygosity of domesticated strains (ranged 0.697–0.804) was also lower compared to the wild population (0.952). Estimated Ne also decreased in all domesticated strains (ranged 10.3–126.0) compared to the wild population (1422.5). The UPGMA tree and 3‐D FCA showed that there were two main clusters containing domesticated strains, and the wild population was at the middle of both of the domesticated clusters. The STRUCTURE analysis also supported the phylogenetic analysis, and revealed three sub‐clusters in the domesticated strains. Pairwise F_ST revealed that all domesticated strains were statistically different from the wild population, and also the differentiation between domesticated strains was all statistically significant. Information on genetic diversity and structure of domesticated strains of red sea bream obtained in this study will be useful for future broodstock management and selective breeding programmes.     

Effects of dietary taurine levels on epidermal thickness and scale loss in red sea bream,Pagrus major

Taurine is often added to artificial fish diets to compensate for a reduction in fish meal (FM). However, the taurine content of FM‐based diets is typically lower than in diets consisting of raw fish, even in diets where FM is the only protein source. We evaluated the effects of dietary taurine in FM‐based diets on epidermal thickness and scale detachability in red sea bream Pagrus major. We compared the effect of diets containing 0% (control), 0.3% (Tau‐0.3%), 0.6% (Tau‐0.6%) and 1.0% (Tau‐1.0%) taurine. Red sea bream (average body weight, 39 g) were fed these diets for 7 weeks. Taurine supplementation had no effect on growth, feed intake, feeding efficiency, or survival. However, the epidermal thickness was higher in fish in the Tau‐0.6% and Tau‐1.0% groups than in the control and Tau‐0.3% groups. Similarly, scale loss was significantly higher in the control group than in the Tau‐0.6% and Tau‐1.0% groups. Our results suggest that supplementation with >0.6% taurine (1.0% in diet) improves skin condition.     

Purification and characterization of phospholipase A2 from the pyloric caeca of red sea bream, Pagrus major

A phospholipase A₂ was purified 55,000-fold in a yield of 10% from the lipid-free extract of powder of the pyloric caeca of red sea bream to near homogeneity by sequential column chromatography on S-sepharose fast flow, butyl-cellulofine, Asahipak ES-502C cation-exchange HPLC, TSK gel G3000SW gel-filtration HPLC, and Asahipak ODP-50 reversed-phase HPLC. The final preparation showed a single band with the apparent molecular mass of 14 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and an estimated specific activity was 717 µmol min^-1 mg^-1 protein. The purified enzyme had a pH optimum in the range of pH 8.0–9.0 and required the presence of both 8 mM of Ca²⁺ and from 2 to 10 mM of sodium deoxycholate for its maximal activity, using 2 mM of phosphatidylcholine as a substrate. The purified enzyme preferentially hydrolyzed the 2-acyl ester bonds of both phosphatidylglycerol and phosphatidylcholine in the presence of sodium deoxycholate, followed in order by phosphatidylethanolamine and phosphatidyl-serine. In contrast to porcine pancreatic PLA₂, pyloric caeca PLA₂ hydrolyzed mixed-micellar phosphatidylcholine substrate effectively, regardless of the kinds of bile salts used. These results indicate that Ca²⁺-dependent low molecular mass PLA₂, so called secretory PLA₂, occurs in the pyloric caeca of red sea beam.     

Efficacy of taurine supplementation for preventing green liver syndrome and improving growth performance in yearling red sea bream Pagrus major fed low-fishmeal diet

ABSTRACT:   This study was performed to evaluate the efficacy of taurine supplementation for preventing green liver syndrome and improving growth performance in red sea bream Pagrus major fed a low-fishmeal (FM) diet. Yearling red sea bream were fed for 34 weeks on low-FM diets either supplemented with taurine, or without taurine, and the tissue taurine and bile pigment concentrations were measured. Compared to the fish fed the FM diet, fish fed the low-FM diet without taurine supplementation resulted in inferior feed performances and higher incidence of green liver related to the morphological transformation of the erythrocytes. In these fish, the hepatopancreatic taurine concentration was significantly lower and hepatopancreatic biliverdin concentration was high compared to the fish fed the FM diet. These parameters were markedly improved by taurine supplementation of the low-FM diet and were similar in levels to the fish fed the FM diet. These results indicate that green liver appearance and inferior feed performances of red sea bream fed the low-FM diet without taurine supplementation were caused by dietary taurine deficiency, and indicate the requirement of taurine supplementation to low-FM diets for red sea bream.     

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.