Studies on the green liver in cultured red sea bream fed low level and non-fish meal diets: Relationship between hepatic taurine and biliverdin levels

SUMMARY: The aim of this study was to investigate the cause of green liver in red sea bream fed substitute protein diets. Red sea bream Pagrus major was given either of the following diets for 28 weeks: (1) control diet (50% fish meal), (2) low level fish meal diet (15% fish meal), and (3) non-fish meal diet (0% fish meal). The green liver was observed in all groups tested, but the incidence was much higher in the experimental diet groups. The feeding of substitute protein diets reduced plasma triglyceride and cholesterol levels. However, there was no significant difference in plasma hepatic enzyme activities and plasma bile salts concentration among the treatments. Fish fed the substitute protein diets showed low hepatic taurine levels with an appearance of a biliverdin in the liver. Moreover, the proportion of ditaurobilirubin to total biliary bile pigments was significantly lower in fish fed the substitute protein diets. These data indicate that feeding of substitute protein diets did not induce any cholestatic hepatobiliary obstructions and that the low hepatic taurine level was one of the probable factors responsible for the occurrence of green liver in red sea bream fed substitute protein diets.     

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.     

Necessity of dietary taurine supplementation for preventing green liver symptom and improving growth performance in yearling red sea bream Pagrus major fed nonfishmeal diets based on soy protein concentrate

The necessity of dietary taurine supplementation for preventing green liver symptom and improving growth performance of red sea bream Pagrus major fed nonfishmeal (non-FM) diets was investigated. Yearling red sea bream (initial body weight, 580 g) were fed for 36 weeks on non-FM diets based on soy protein concentrate (SPC) supplemented with taurine at levels of 0%, 0.5%, 1.0%, 1.5%, and 2.0%. Specific growth rate (SGR) and feed conversion ratio (FCR) of fish fed the taurine-unsupplemented SPC diet were markedly inferior. In these fish, incidence of green liver was markedly higher and was accompanied by a decrease of tissue taurine concentration and an increase of hepatopancreatic bile pigment content. The green liver symptom was mainly caused by an increase of hemolysis since the erythrocytes became osmotically fragile due to taurine deficiency. Physiological abnormality and growth performance (SGR and FCR) were markedly improved by taurine supplementation to the SPC diets. These results indicate that dietary taurine supplementation is necessary for yearling red sea bream fed non-FM diet based on SPC to maintain normal physiological condition and growth performance.     

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.     

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.     

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.     

Effects of feeding a fish meal‐free soy protein concentrate‐based diet on the growth performance and nutrient utilization of red sea bream (Pagrus major)

Two groups of red sea bream (average body weight 85 g) were fed to satiation fish meal (FM) or FM‐free soy protein concentrate (SPC)‐based diet (FM‐sat and SPC groups respectively). A third group (FM‐pair) was fed the same amount of FM as of SPC fed to SPC‐sat group for 6 weeks. Growth rate of SPC group was significantly slower than of FM‐sat and FM‐pair groups (p < 0.05). Feed conversion ratios of FM‐sat and FM‐pair groups were similar and were better than that of SPC group (p < 0.05). Visceral fat somatic index of SPC group was significantly lower than of the other groups (p < 0.05), while that of the latter two groups were similar. Plasma triglyceride of FM‐sat and FM‐pair groups was similar and lower than the level of SPC group (p < 0.05). Trypsin activity in hepatopancreas of SPC group was significantly lower than that of other groups (p < 0.05). We suggest a low growth of SPC group was not simply related to low‐feed intake. Feeding of a FM‐free SPC‐based diet may reduce lipid accumulation and consequently lead to low‐growth performance, in red sea bream.     

Interactive effects of dietary vitamin C and phospholipid in micro‐bound diet for growth,survival, and stress resistance of larval red sea bream,Pagrus major

This study was conducted to examine the effects of dietary ascorbic acid (AsA) and phospholipid (PL) and their interaction on growth, survival, and stress resistance in red sea bream larvae. Twenty‐six days old red sea bream were fed nine micro‐bound diets supplemented three levels of AsA (0, 800 and 1600 mg kg^?1 diet) and PL (0, 20 and 40 g kg^?1 diet) for 15 days. Dietary AsA and PL were both significant factors on survival rates. There was also an interaction between dietary AsA and PL on survival rate (P < 0.05). The larvae fed 800 or 1600 mg kg^?1 AsA with 40 g kg^?1 PL diets showed the highest survival rate, with values similar to those of the live‐food supplemented group. Stress resistance against low salinity exposure significantly increased with increased dietary level of AsA and PL. However, significant interaction of AsA and PL was not detected. The larvae fed 1600 mg kg^?1 AsA with 40 g kg^?1 PL diet showed the highest stress resistance among all diets, but it was not significantly different than that of larvae fed 800 mg kg^?1 AsA with 40 g kg^?1 PL diet. This study clearly demonstrated that combined use of AsA and PL can improve survival of 26–40 days posthatching red sea bream larvae. Moreover, the present study suggested that 800 mg kg^?1 AsA with 40 g kg^?1 PL in diet was needed for producing high quality seedling under the stressful conditions.     

Dietary inorganic phosphorus or microbial phytase supplementation improves growth,nutrient utilization and phosphorus mineralization of juvenile red sea bream,Pagrus major,fed soybean‐based diets

A 2 × 3 factorial design with triplicates examined the interaction between dietary inorganic phosphorus (IP) and phytase on growth, mineral utilization and phosphorus (P) mineralization in juvenile red sea bream. The treatments were three levels of dietary IP supplementation at 0, 2.5 and 5 g kg^?1, either without or with phytase supplementation [2000 FTU kg^?1; phytase unit is defined as the amount of enzyme activity which liberates 1 micromol of inorganic phosphorus per minute at pH 5.5 and 37 °C at a substrate concentration (sodium phytate) of 5.1 mmol L^?1]. Juvenile red sea bream (IBW = 1.3 g ± 0.1) were stocked twelve fish per tank and fed for 50 days. Growth and feed efficiency were significantly (P < 0.05) enhanced by both dietary P and phytase supplementation. Feed intake and survival rate were not significantly affected by the dietary treatments. Both dietary IP and phytase supplementation significantly increased plasma IP and Mg levels. Concentration of vertebral mineral and scale P was significantly increased by both dietary treatments. A skeletal malformation syndrome of scoliosis occurred in fish fed both non‐IP and non‐phytase supplemented diet. Interaction between main dietary effects was detected for vertebral Zn, scale P and whole‐body ash and Mg content. With regard to growth and other examined productivity traits, phosphorus requirement of juvenile red sea bream can be met if supplemented with 2000 FTU phytase kg^?1 or in the absence of phytase, by dietary inclusion of 2.5–5 g kg^?1 of IP.     

Effects of natural feeding stimulants and glutamic acid supplementation on the feed intake,growth performance and digestive enzyme activities of red sea bream (Pagrus major) fed fish meal‐free soy protein concentrate (SPC)‐based diet

Two experiments were conducted for red sea bream (Pagrus major). In experiment 1, the optimum level of glutamic acid and natural feeding stimulants to enhance feed intake were determined and found that glutamic acid level of 0.5% and fish meat hydrolysate (FMH) were effective. In experiment 2, fish were fed with soy protein concentrate (SPC)‐based diet with synthetic feeding stimulants (Basal diet), the Basal diet with FMH (FMH diet), the FMH diet with glutamic acid (FMHG diet) and with fish meal diet (FM diet) as a control until satiation for 8 weeks. Feed intake of FMHG‐fed fish was significantly higher than others (p < 0.05). Specific growth rate and the feed conversion ratio of FMHG were comparable to those of FM‐fed fish (p > 0.05). Relative visceral fat ratio and crude lipid content of any SPC‐based diet‐fed fish tended to be lower than those of FM diet‐fed fish. There were no significant differences in trypsin and lipase activities hepatopancreas among treatments. SPC can be utilized as a sole protein source in a diet for red sea bream. The lower growth performance in SPC‐based diet‐ fed fish was not due to poor digestive enzyme secretion but could be associated with lipid utilization disorder.     

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.     

The microhabitat and size of gammarid species selectively predated by young red sea bream Pagrus major

SUMMARY: Mechanisms of prey selection by young red sea bream Pagrus major were examined in Shijiki Bay by comparing the species composition of gammaridean amphipods in the diet with that in the environment. Although gammarids were the major source of food for young red sea bream, their species composition in fish stomachs changed with the growth of fish from June to October and always differed from that in the environment. The patterns of selection of gammarids were related most closely to the microhabitat of each species before mid-July. The order of selectivity of species in terms of the microhabitat was as follows: epifaunal species > shallow-burrowing species > infaunal tube-dwelling species > deep-burrowing species. After mid-July, however, selection was determined by the body size of gammarid species. Byblis japonicus that were large enough as a prey species were mainly consumed and other smaller species were not selectively eaten. Thus, the microhabitat and the body size of gammarids play dominant roles in prey selection by young red sea bream.     

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.     

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.     

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.