• Volume 40,Issue 9,2016 Table of Contents
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    • Changes on the contents of total volatile basic nitrogen and biogenic amines in two common species of freshwater fish, Hemiculter leucisculus and Erythroculter erythropterus, at high temperature storage

      2016, 40(9):1505-1511. DOI: 10.11964/jfc.20160310312

      Abstract (1855) HTML (477) PDF 758.75 K (1737) Comment (0) Favorites

      Abstract:Hemiculter leucisculus and Erythroculter erythropterus are small fishes which are common freshwater species in China. They are often fed to aquatic animals, and are also food for humans. In this study, the total volatile basic nitrogen (TVBN) and nine biogenic amines (cadaverine, putrescine, histamine, tyramine, serotonin, spermidine, spermine, dopamine and octopamine) of H.leuciclus and E.erythropterus at 30℃ storage were examined in every 6h for 48h. The correlation analysis of the contents of TVBN or biogenic amines with the storage time was also evaluated for these two species. The results showed that there were differences in the contents of biogenic amines and TVBN in H.leucisculus and E.erythropterus. The TVBN contents of H.leuciclus and E.erythropterus obviously increased with the storage time increasing. The TVBN content of E.erythropterus was significantly higher than that in H. Leucisculus at all time except at 42 h (P<0.05). The TVBN contents of H. leucisculus and E.erythropterus exceeded the national edible standard of 30 mg/100 g after 12h. Therefore these two species were not suitable to eat after 12 h. The four main biogenic amines detected in these two species were cadaverine, putrescine, tyramine and histamine. The contents of these four biogenic amines increased significantly with the extension of storage time (P<0.05). The contents of cadaverine and histamine in E.erythropterus were significantly higher than those in H.leuciclus (P<0.05). The contents of TVBN, cadaverine, putrescine, histamine and tyramine were found to correlate closely (P<0.01) with the storage time in these two species, and the same situation was also found for serotonin from E.erythropterus.

    • Effects of replacing fish meal with fermented mulberry leaves on the growth, lipid metabolism and antioxidant capacity in largemouth bass (Micropterus salmoides)

      2016, 40(9):1408-1415. DOI: 10.11964/jfc.20150609909

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      Abstract:This experiment was conducted to study the effect of replacement of fish meal by fermented mulberry leaves on the growth, lipid metabolism and antioxidant capacity of largemouth bass[with the initial weight (10 g)]. Four isonitrogenous and isoenergetic diets (CP 42%, GE 18 MJ/Kg) were formulated to contain different levels of fermented mulberry based upon the basal diet to replace fish meal 0 (D1), 15% (D2), 30% (D3) and supplement EAA based on 30% (D4). After 8 weeks, the results showed that final weight (FW), specific growth rate (SGR), viscera somatic index (VSI) and hepatopancreas somatic index (HSI) were decreased significantly when the replacement ratio of fish meal by fermented mulberry was 30%, and supplement crystalline amino acids (CAA) could improve the growth of largemouth bass obviously. There were no significant differences in protein efficiency ratio (PER), feed conversion ratio (FCR), feeding rate (FR), condition factor (CF) and whole body general composition for each test group. The largemouth bass serum of CHO, TG, HDL-C and LDL-C were decreased observably but the ratios of HDL-C/CHO and HDL-C/LDL-C showed the opposite trend. The LDL-C decreased as fermented mulberry ratio increased. Superoxide dismutase (SOD) activities increased significantly as fermented mulberry ratio increased, but malondialdehyde (MDA) decreased markedly. Results of above show that a suitable percentage of fermented mulberry replacement of fish meal has no effect on the growth of largemouth bass. Too high replacement levels would inhibit the growth of largemouth bass, but could improve the lipid metabolism and antioxidant capacity of largemouth bass. Supplement of essential amino acids can improve the utilization of fermented mulberry, and promote the growth of largemouth bass.

    • Effects of dietary silymarin on growth performance, enzymes of hepatic lipid metabolism and antioxidant ability in GIFT Oreochromis niloticus

      2016, 40(9):1309-1320. DOI: 10.11964/jfc.20160210292

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      Abstract:Silymarin was extracted from Silybum marianum (St. Mary's thistle, milk thistle), and has been used to treat liver, spleen and gallbladder disorders due to functions in anti-radical, antioxidant, anti-lipid peroxidation, anti-inflammatory and to protect the stability of plasma membrane. However, it remains unknown whether silymarin has the same functions in juvenile GIFT Oreochromis niloticus. A 9-week feeding experiment was performed to investigate the effects of dietary silymarin levels on growth performance, hepatic lipid metabolism enzymes activities and antioxidant ability in juvenile GIFT[initial body weight (8.17±0.31) g]. Five practical diets were formulated with incorporation of silymarin at the levels of 0 (control), 100, 200, 400 and 800 mg/kg. At the end of the feeding trial, growth performance, body composition, serum biochemical indices, hepatic lipid metabolism enzymes activities and antioxidant enzymes activities were measured. The results were as follows:① Compared with those in the other groups, the highest weight gain rate (WGR), specific growth rate (SGR) and condition factor(CF) were observed in fish fed the diets containing 100 mg/kg silymarin. By contrast, feed conversion rate(FCR) of the fish fed the diets containing 100 mg/kg silymarin was the lowest and significantly decreased compared with control group. Dietary silymarin supplementation significantly decreased hepatosmatic index (HSI) and viscerosomatic index(VSI). There was no significant difference of survival rate among all the groups. ② The lipid content in whole body, muscle and liver was the lowest at the group of 100 mg/kg diet and significantly lower than the control group. Dietary silymarin supplementation significantly decreased triglyceride in serum. The activities of serum aspartate transaminase (AST) and alanine aminotransferase (ALT) were both decreased initially and then increased as silymarin level increased, and the lowest activity of AST and ALT were observed in the group fed 200 and 100 mg/kg diet, respectively. ③ For lipid metabolism enzymes, dietary silymarin supplementation significantly elevated the activities of lipoprotein lipase (LPL) and total lipase (TL) in liver, and the highest activity of lipase (LPS) was in the group of 200 mg/kg diet, while it was significantly lower than the control group. ④ Feed added silymarin significant increased glutathione peroxidase (GSH-PX) activity and decreased catalase (CAT) activity in liver, and significantly reduced malondialdehyde (MDA) content in liver. ⑤ Liver slices of GIFT treated with dietary silymarin addition at 100 mg/kg showed few hepatocytes with empty vacuoles and less nuclear migration. In conclusion, appropriate dietary silymarin supplementation could promote growth performance, decrease muscle, whole body and hepatopancreas lipid contents, and regulate lipid metabolism enzymes and antioxidant ability in liver. On the basis of the above results, the proper dose of silymarin in juvenile GIFT diet was recommended to be 100 mg/kg.

    • Effect of dietary astaxanthin on culture-based dominant bacterial community isolated from the intestinal tract and gills of male Chinese mitten crab (Eriocheir sinensis)

      2016, 40(9):1416-1430. DOI: 10.11964/jfc.20160210289

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      Abstract:Chinese mitten crab (Eriocheir sinensis) is one of the important economic species of freshwater aquaculture in China. Dietary astaxanthin affects the number and composition of culture-based dominant bacteria from intestinal tract and gills, which is closely linked to the body's immunity. In order to realize the relationship between dietary astaxanthin and the balance of bacteria in intestinal tract and gills, five diets with various concentration astaxanthin (0.00, 26.60, 41.62, 81.37 and 75.35 mg/kg, respectively) were fed to adult male crabs for 70 days. The dominant bacteria number and composition of five crab groups were measured by culture-based enumeration techniques, and then the isolates were sequenced by 16S rRNA gene analysis. A total of 106 bacterial strains were isolated from the intestinal tract, gills and aquaculture water. 92 bacterial strains were dominant bacteria, of which 29 bacterial strains were from intestinal tract, 32 bacterial strains from gills and 31 bacterial strains from aquaculture water. At the genus level, the dominant bacteria were Citrobacter, Pseudomonas and Aeromonas in the intestinal tract, by contrast, the dominant bacteria were Citrobacter, Bacillus, Pseudomonas and Aeromonas in the gill, and the dominant bacteria were Bacillus, Aeromonas, Citrobacter, Pseudomonas and Chryseobacterium in the aquaculture water. The total number of culture-based dominant bacteria in intestinal tract was the highest (1.06×108 cfu/g) in group 2. The number of potential pathogenic bacteria of group 5 (75.35 mg/kg) was significantly decreased in intestinal tract. However, the total number of dominant bacteria of group 5 was significantly higher than the other four groups in the gill, and the numbers of dominant bacteria had no significant difference in all five groups in aquaculture water. To conclude, dietary astaxanthin with various concentrations could significantly affect the culture-based dominant bacteria composition of intestinal tract and gill in male E. sinensis. The number of potentially pathogenic bacteria in intestinal tract and gills accounted for high proportion in group 3. This study for the first time analyzed the effect of astaxanthin on number and composition of culture-based dominant bacteria of intestinal tract and gills in male Chinese mitten crab, and discussed the relationship between concentration of dietary astaxanthin, aquaculture water and the number of culture-based dominant bacteria from intestinal tract and gill, and the results may provides a theoretical basis for E. sinensis diet exploitation and bacterial flora control in the future.

    • Effects of dietary tert-butylhydroquinone on growth performance, blood biochemical parameter, non-specific immunity and intestinal tissue structure of turbot (Scophthalmus maximus)

      2016, 40(9):1321-1329. DOI: 10.11964/jfc.20160310303

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      Abstract:A feeding trial was conducted to evaluate the effects of dietary tert-butylhydroquinone (TBHQ) on growth performance, blood biochemical parameter, non-specific immunity and intestinal tissue structure of turbot (Scophthalmus maximus). Four isonitrogenous and isoenergetic diets were formulated to contain graded levels (0, 150, 450 and 750 mg/kg) of TBHQ. Fish[initial average weight (8.31±0.04) g] were randomly allocated to 24 tanks, and each tank was stocked with 30 fish. Fish were fed twice daily to apparent satiation for 12 weeks. The results indicated that fish fed the diets containing 450 and 750 mg/kg TBHQ had significantly lower weight gain ratio and specific growth rate than that of fish fed the basal diet (the control group). Fish fed the diet containing 450 mg/kg TBHQ had significantly higher activity of serum alkaline phosphatase than that of fish fed the diet containing 150 mg/kg TBHQ. Fish fed the diet containing 150 mg/kg TBHQ had significantly lower albumin and high density lipoprotein content than that of fish fed the basal diet and the diet containing 450 mg/kg TBHQ; fish fed the diet containing 450 mg/kg TBHQ had significantly higher serum creatinine content than that of fish fed the basal diet; fish fed the diet containing 150 mg/kg TBHQ had significantly lower serum total protein content than that of fish fed the basal diet. Fish fed the diets containing 450 and 750 mg/kg TBHQ had significantly higher activity of serum CAT, lysozyme activity, and fish fed the diet containing 750 mg/kg TBHQ had significantly higher respiratory burst activity of renal phagocytes. Fish fed the diet containing 750 mg/kg TBHQ had significantly lower serum SOD activity. Fish fed the diets containing 450 and 750 mg/kg TBHQ had significantly lower ratio of intestinal villus length to bowel diameter than that of fish fed the basal diet, and fish fed diet containing 750 mg/kg TBHQ had significantly lower ratio of microvillus length to bowel diameter than that of fish fed the basal diet. In conclusion, the results of the present study indicated that 150 mg/kgTBHQ in diets of juvenile turbot had no significant effects on weight gain, physiological or biochemical indexes, while fish fed diet with more TBHQ (>450mg/kg) had negative effects on the growth and physiological status.

    • Effect of dietary taurine supplementation on growth, intestine structure and resistance to acute crowding stress in juvenile black carp (Mylopharyngodon piceus) fed low fish meal diets

      2016, 40(9):1330-1339. DOI: 10.11964/jfc.20151210200

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      Abstract:An 8-week feeding experiment was conducted to investigate the effects of dietary graded level of taurine supplementation on growth performance, intestine structure and resistance to acute crowding stress of juvenile black carp (Mylopharyngodon piceus) (initial mean body weight 5.90±0.03 g). The diet containing 20% fish meal was set as the control group. All the other groups contained 10% fish meal and were supplemented with 0% (I0), 0.05% (I0.05), 0.1% (I0.1), 0.2% (I0.2) and 0.4% (I0.4) taurine, respectively. Each diet was randomly fed to triplicate groups of 30 fishes per tank (300 L). After the feeding experiment, according to growth performance, a 24 h acute crowding stress was conducted in 4 group (the control, I0, I0.1, I0.4, and the density is 100 g/L. The results showed that the weight gain rate (WGR) of black carp in I0 group was significantly lower than that in the control group (P<0.05). At the lower fish meal level, WGR of black carp first rose and then decreased with increasing dietary taurine supplementation level, and the maximum value was found in 0.1% group (P<0.05), who had no significant difference with the control group. Compared with the control, the villus height of the I0 group were higher, crypt depth and number of goblet cells of the I0 group were smaller; At the lower fish meal level, all fish fed the diets with dietary taurine supplementation had a greater crypt depth, goblet cell and lymphocytes number. Moreover, a lower villus height, villus height, crypt depth, goblet cell and lymphocytes number of intestine in the I0.1 group were significantly higher than the control group (P<0.05). As the growth of the crowded stress time, cortisol, blood glucose (GLU) and lysozyme (LSZ), complement C3, superoxide dismutase (SOD) and glutathione (GSH) all had a trend of first rose and then decreased, and the maximum were found at 2 h and 8 h. There were different increase of these indexes among dietary treatments. The serum glucose and cortisol of fish fed the diet with 10% fish meal level were significantly higher than those in the other group (P<0.05), but serum LSZ, complement C3, SOD and GSH in I0 group lower than those in the other group (P>0.05). These results suggested that dietary taurine supplementation can enhance growth performance of black carp, improve intestine structure, and increase the ability of black carp's resistance to acute crowding stress.

    • In vivo absorption and release characteristics of Pb and Cd in diet in Eriocheir sinensis

      2016, 40(9):1431-1439. DOI: 10.11964/jfc.20160110226

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      Abstract:To learn more about the enrichment and release characteristics of heavy metals in the feed and the tissues of Eriocheir sinensis, kinetic parameters were gained for biological accumulation and release of two heavy metals Pb and Cd in gill, hepatopancreas and muscle of E. sinensis, by using two-compartment bioconcentration model to carry out the experiment in which the crabs were exposed to the feed with Pb concent of 10.21 mg/kg, 22.01 mg/kg, 40.81 mg/kg, and Cd concent of 1.78 mg/kg, 2.80 mg/kg, 4.48 mg/kg, so as to provide theoretical basis for the distribution, migration and enrichment, and provide guiding significance for safe production eriocheir sinensis for Pb and Cd in E. sinensis. Heavy metal uptake rate constant (k1), extracting rate constant (k2), bioconcentration factor (BCF) and biological half-lives (B1/2), equilibrium concentration of heavy metals in vivo CAmax were obtained by non-linear curve fitting. The results showed:(1) The concent of Pb in crab gill, hepatopancreas and muscle exhibited an apparently positive correlation to the accumulation time as well as the concent. On the forty-eighth day of enrichment, the content of Pb in each tissue reached the maximum, the content in gill was 0.18 mg/kg, 1.14 mg/kg, 1.27 mg/kg and 1.91 mg/kg respectively; Content in hepatopancreas was 1.00 mg/kg, 2.17 mg/kg, 2.33 mg/kg and 3.50 mg/kg respectively; Content in muscle was 0.18 mg/kg, 0.73 mg/kg, 1.00 mg/kg and 1.35 mg/kg respectively; The accumulation of Cd in gill and hepatopancreas presented the similar results to that of Pb.On the forty-eighth day of enrichment, the content of Cd in gill reached the maximum, the content of was 0.026 mg/kg, 0.073 mg/kg, 0.107 mg/kg and 0.154 mg/kg respectively. In addition to the twenty-fourth day of the group C, the content of the hepatopancreas reached the highest,was 1.90 mg/kg; The other 3 groups of experiments, the highest content was reached in the enrichment of forty-eighth days, it was 0.33 mg/kg, 1.05 mg/kg and 1.24 mg/kg respectively. In group C, the content of forty-eighth days decreased, it wsa 1.76 mg/kg. But there was no obvious rule of Cd in muscle. (2) The accumulation and release of Pb and Cd were both slow in tissues and organs of E. sinensis. At steady state the concentration of Pb in crab gill, hepatopancreas and muscle ranged from 1.07 mg/kg to 1.69 mg/kg, 4.87 mg/kg to 4.95 mg/kg, 0.79 mg/kg to 1.28 mg/kg, the concentration of Cd in crab gill, hepatopancreas ranged from 0.06 mg/kg to 0.14 mg/kg, 1.25 mg/kg to 2.66 mg/kg. The bioconcentration factor (BCF) of Pb and Cd in the tissues and organs ranged from 0.03 to 0.48 and 0.03 to 0.87, respectively, indicating that the absorption of Cd was much more higher than that of Pb in E. sinensis. The biological half-lives of Pb and Cd in the tissues and organs (B1/2) ranged from 9 to 67 d and 8 to 48 d, which signified that the release of Cd is significantly slower than that of Pb. (3) Pb and Cd were accumulated selectively in the major organs and tissues in E. sinensis. After feeding with the feed, Pb and Cd were distributed in the same way, namely, hepatopancreas > gills > muscle.

    • Effect of bile acid level in high lipid diet on the intestinal structure and lipid metabolic enzymes activities of juvenile Schizothorax prenanti

      2016, 40(9):1340-1348. DOI: 10.11964/jfc.20160110268

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      Abstract:This experiment was conducted to examine the effect of high lipid diet on the intestinal structure and lipid metabolism enzymes in juvenile Schizothorax prenanti treated with bile acid. In our study, a total of 360 juvenile S. prenanti, with average body weight of (12.74±0.14) g, were randomly distributed into four groups with three replicates each. The high lipid diet diets with different bile acid levels (0, 75, 150 and 300 mg/kg) were fed to juvenile S. prenanti for 70 days. The results showed that the foregut and midgut intestinal tube wall thickness, as well as villi length significantly increased with increasing dietary bile acid levels up to 75 mg/kg diet, and remained nearly constant thereafter. Meanwhile, the foregut plica width was significantly reduced with increasing dietary bile acid levels up to 75 mg/kg diet, where the response reached a plateau. In addition, we found that the intestinal lipase (LPS), lipoprotein lipase (LPL), hepatic lipase (HL) and total lipase (TL) increased firstly and then tended to stabilize. Interestingly, the activities of LPS and LPL of juvenile S. prenanti were the highest for fish fed with diet containing 300 mg/kg bile acid diet, which were 2881.17 U/g and 14.43 U/mg prot, respectively. While the activities of HL and TL were highest for fish fed 150 mg bile acid/kg diet, which were 43.70 and 58.03 U/mg prot, respectively. Collectively, the results indicated that 75-150 mg bile acid/kg diet can improve the fat metabolism enzyme activity, promote the utiligation of diet lipid and improve intestinal structure, and protect the intestinal health in juvenile S. prenanti.

    • Effect of fasting on body composition, muscle fatty acid profiles and serum biochemical parameters of large yellow croaker (Larimichthys crocea)

      2016, 40(9):1440-1450. DOI: 10.11964/jfc.20151210216

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      Abstract:This study was conducted to determine the effect of fasting on body composition, muscle fatty acid profiles and biochemical parameters of serum in large yellow croaker (Larimichthys crocea). 105 net-cage cultured healthy fish with similar body weight (249.07±7.28 g) were selected for fasting experiment. The muscle and blood were sampled from experimental fish after 0, 3, 7, 14, 21, 28, 35, 42, 49 days of fasting for chemical analysis, respectively. The results showed that moisture and crude ash of muscle increased and crude fat decreased during fasting period. The protein content of muscle significantly decreased at 21 days of fasting and then remained constant. Saturated fatty acid (SFA) and monounsaturated fatty acid (MUFA) content in muscle showed no significant change(P>0.05), while the total polyunsaturated fatty acid (PUFA) and n-3 high unsaturated fatty acid (n-3HUFA) content significantly decreased at the end of experiment. The activities of serum alanine transaminase (ALT), glutamic-oxaloacetic transaminase (AST), lactate dehydrogenase (LDH) and the contents of serum glucose (GLU), Kalium (K+), sodium (Na+), calcium (Ca2+) and chloride (Cl-) significantly increased, while serum triglyceride (TG), Cholesterol (CHOL) contents significantly reduced after 3 days of fasting. Nevertheless serum total protein (TP), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) contents and alkaline phosphatase (ALP) activity keep relatively stable(P>0.05). The content of GLU, protein, lipid and metabolic enzymes activities in serum fluctuated decrease with prolongation of fasting, while Na+ and Cl-content fluctuated increase. The results suggested that large yellow croaker main used body lipid as energy source during fasting period. The nutrient metabolism intensity showed wave-like fluctuated and turn weak as fasting time going by. But the immunity and metabolism function of large yellow croaker were not significantly harmed by long time fasting.

    • Effects of different dietary pantothenic acid levels on growth performance, feed utilization and serum indices of juvenile Litopenaeus vannamei

      2016, 40(9):1349-1358. DOI: 10.11964/jfc.20160110258

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      Abstract:An 8-week feeding trial was conducted to evaluate the effects of different dietary pantothenic acid levels on growth performance, feed utilization and serum indices of juvenile Pacific white shrimp (Litopenaeus vannamei). Six isonitrogenous and isolipidic practical diets (41% crude protein and 8% crude lipid) were formulated to supplement with pantothenic acid levels were of (control diet), 50, 100, 150, 300 and 600 mg/kg, respectively. The analyzed values of dietary pantothenic acid are 20.9, 69.3, 99.0, 150.2, 304.4 and 513.6 mg/kg, respectively. Each diet was randomly assigned to 3 replicates of 40 juvenile shrimps (initial weight approximately 0.73±0.12 g). The results indicated that weight gain rate and specific growth rate were not significantly affected by dietary pantothenic acid levels (P>0.05). However, shrimp fed the control diet had lower survival rate than those fed the diets supplemented with pantothenic acid (P<0.05), while there were no significant differences among those supplemented with pantothenic acid. Feed efficiency and protein efficiency rate significantly increased with dietary pantothenic acid level increasing from 20.9 mg/kg to 99.0 mg/kg, then decreased with further increase of dietary pantothenic acid level from 150.2 mg/kg to 513.6 mg/kg. Composition of whole body and muscle in the juvenile L. vannamei were not significantly influenced by the dietary pantothenic acid levels (P>0.05). Shrimp fed control pantothenic acid diet had lower glucose concentration in serum than those fed the other diets, while there were no significant differences among all diets supplemented with pantothenic acid. Shrimp fed the diet containing 150.2 mg/kg pantothenic acid had lower TG in serum than those fed the diets containing 20.9 mg/kg and 513.6 mg/kg. Total protein and cholesterol in serum were not affected by dietary pantothenic acid levels. SOD activity in serum increased significantly with dietary pantothenic acid level increasing from 20.9 mg/kg to 150.2 mg/kg, then decreased significantly with further increase of dietary pantothenic acid level (P<0.05). Shrimp fed the diet containing 99.0 mg/kg pantothenic acid had higher T-AOC than the other diets. Shrimp fed the diet containing 69.3 mg/kg pantothenic acid and control diet had higher level MDA in serum than those fed the other diets, and the lowest MDA in serum occurred at diet containing 150.2 mg/kg pantothenic acid (P<0.05). Based on a two-slope broken-line model between feed efficiency, productive protein value and dietary pantothenic acid levels, the optimal dietary pantothenic acid requirement is estimated to be 113.40 and 119.87 mg/kg for juvenile Pacific white shrimp, respectively.

    • Effects of salinity-light intensity-temperature combinations on the growth and highly unsaturated fatty acids of Chaetoceros sp. SHOU-B98

      2016, 40(9):1451-1461. DOI: 10.11964/jfc.20150810040

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      Abstract:Orthogonal experiments L9(34) were applied to investigate the effects of different salinity (6, 12 and 18), light intensity[10, 30 and 50 μmol/(m2·s)] and temperature (10, 20 and 30℃) combinations on the growth and cellular contents of arachidonic acid (ARA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in Chaetoceros sp. SHOU-B98. The results showed that salinity, light intensity, temperature and the interaction of the three factors had significant effects on growth and highly unsaturated fatty acid contents of the microalgae. Under salinity 12, light intensity 30 μmol/(m2·s) and temperature 30℃ combination and salinity 18, light intensity 50 μmol/(m2·s) and temperature 20℃ combination, the microalgae displayed the fastest growth and highest biomass since the 4th day (P<0.05). The best culture conditions for cell to accumulate different highly unsaturated fatty acids were also different. The optimal combined culture condition for Chaetoceros sp. SHOU-B98 cell to accumulate ARA was combination of salinity 6, light intensity 50 μmol/(m2·s) and temperature 30℃. While the optimal culture condition to accumulate EPA or DHA was combination of salinity 18, light intensity 50 μmol/(m2·s) and temperature 20℃. Therefore the suitable culture condition for microalgae Chaetoceros sp. SHOU-B98 with fast growth and high fatty acid nutrition value is the combination of salinity 18, light intensity 50 μmol/(m2·s) and temperature 20℃.

    • Effects of dietary methionine levels on muscle amino acids composition and serum antioxidant ability of GIFT Oreochromis niloticus

      2016, 40(9):1359-1367. DOI: 10.11964/jfc.20150109700

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      Abstract:A feeding trial was conducted to evaluate the effects of dietary methionine levels on muscle amino acids composition and serum antioxidant ability of GIFT tilapia (Oreochromis niloticus)[initial body weight of (66.76±2.29) g]. Six isonitrogenous and isoenergetic semi-purified diets were formulated to contain 32.09% crude protein,17.82% kJ/g gross energy and six levels of methionine 0.26%, 0.55%, 0.85%, 1.14%, 1.44% and 1.73% respectively (dry matter basis). Each diet was randomly assigned to triplicate groups of 25 fish and fed to apparent satiation by hand thrice daily (8:00, 13:00 and 16:00) for 60 days in indoor farming system. The results showed that the all amino acids content, gross of essential amino acid (ΣEAA), gross of total amino acid (ΣTAA), gross of total delicious amino acid (ΣDAA) in muscle initially increased with increasing dietary methionine levels but then decreased. All the indexes were highest when the dietary methionine level was 1.14% (P<0.05). The ratio of essential amino acids to total amino acids (ΣEAA/ΣTAA), the ratio of delicious amino acid to total amino acids (ΣDAA/ΣTAA), the ratio of branched chain amino acid to aromatic amino acids (ΣBCAA/ΣAAA) had no significant differences (P>0.05). With the increase of dietary methionine level, the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) in serum first decreased, then increased, at the same time, the malondialdehyde (MDA) in serum first increased, then towards stability. Results of above show that the optimum dietary methionine level could effectively improve the muscle amino acid composition and the quality of the muscles, and enhance the serum antioxidant capacity of GIFT tilapia.

    • Prokaryotic expression and bioactivity analysis of orexin A from Cynoglossus semilaevis

      2016, 40(9):1462-1471. DOI: 10.11964/jfc.20151110174

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      Abstract:In order to investigate the underlying mechanisms for food intake regulation of Cynoglossus semilaevis, one of the key feeding factors-orexin A-was produced in vitro in prokaryotic expression system and its possible role in feed intake regulation was explored using in vitro incubation method of hypothalamus gland. The matured peptide fragment of orexin A was amplified and subcloned into the prokaryotic expression vector-pET32a to successfully construct orexin A/pET32a recombinant plasmid which was highly expressed in E.coli BL21 after being induced by IPTG with special fusion polypeptides containing His6 at their N-terminus. The orexin A fusion protein was mainly expressed in supernatant liquid with molecular weight of 24.9 kDa, and the fusion protein products maximally accounted for 52.8% of the whole bacterial protein post 6h induction with 1 mmol/L IPTG at 32℃. Western blotting analysis indicated fusion protein had the antigenicity to His6 antibody. Supernatant after crushing, was purified using Ni2+-NTA affinity chromatography, then the purified protein with molecular weight of 24.9 kDa was obtained. Incubating hypothalamus with recombinant expression orexin A of Cynoglossus semilaevis indicates that orexin A can significantly influence expression of NPY mRNA, orexin A mRNA and secretion of NPY peptide in hypothalamus. Therefore, the obtained recombinant orexin A protein has biological activity in the present study. The present results would be helpful for better understanding the roles of orexin A in feeding regulation and development of high-efficient feeding promotion additives for aquaculture of Cynoglossus semilaevis.

    • Effect of glycyrrhetinic acid (GA) on the physiological responses and immunoreactions of channel catfish (Ictalurus punctatus) injection lipopolysaccharide (LPS)

      2016, 40(9):1368-1375. DOI: 10.11964/jfc.20160410347

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      Abstract:Glycyrrhetinic acid (GA), which is a main active principal constituent of liquorice, is extensively used as antibacterial and anti-inflammatory Chinese medicine. This study was carried out to evaluate the effects of GA on the oxidative stress of Ictalurus punctatus injection lipopolysaccharide (LPS). The fish were randomly divided into two groups:a control group (fed a standard diet) and a treatment group (standard diet supplemented with 0.15 mg/kg GA) and fed for 8 weeks. We then challenged the fish with LPS and recorded changes in red and white blood cell counts (RBC and WBC), plasma cortisol, lysozyme, ACH50, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and hepatic catalase (CAT), superoxide dismutase (SOD) and malondialdehyde (MDA) for a period of 48 h. Supplementation with 0.15 g/kg GA significantly increased plasma lysozyme activity before infection, plasma ACH50 activity at 6 h after infection, WBC hepatic at 6 h after infection, CAT activity 24 h after infection, hepatic SOD activity before and after infection. In addition, the supplemented group had decreased levels of plasma AST and ALT activities 6 h after infection, hepatic CAT 3 h after infection, hepatic MDA 24 and 48 h after infection. Our results suggest that ingestion of a basal diet supplemented with 0.15 g/kg GA prevents LPS-induced immunological challenges response by regulating key enzyme activities, and challenged with LPS (2.0 mg/kg) affect physiological responses and immunoreactions of channel catfish.

    • Studies on larval metamorphosis and feed conversion of Rapana venosa

      2016, 40(9):1472-1478. DOI: 10.11964/jfc.20150910068

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      Abstract:In the past decades, the natural resources of Rapana venosa in China have obviously declined because of over-exploitation and habitat destruction. It is very necessary to study the artificial reproduction of R. venosa for recovering natural resources and meeting the demand of the market. However, large-scale artificial seedling of R. venosa is still hampered by low larval metamorphosis rates and feed conversion. In order to solve these problems, larval rearing, spat collection and juveniles' culture were systematically investigated in the study. The results showed that in the early stage of the planktonic larvae, growth rate of shell height was 21.8 μm/d (0-12 d). In the middle stage of the planktonic larvae, growth rate of shell height was 3 μm/d (13-18 d). In the late stage of the planktonic larvae, growth rate of shell height was 46.8 μm/d (19-30 d). After being induced by the fresh seawater from the precipitation pond, the planktonic larvae began to settle and metamorphose on the first day and the process of settlement and metamorphosis ended with 40% of larval metamorphosis on the fourth day. The juveniles of R. venosa can feed on spats of Crassostrea gigas newly settled to the scallop shells and spats of Scapharca broughtonii newly settled to the monofilament nylon. Growth rate of shell height in the juveniles was 65.6 μm/d (1-8 d). From the ninth day, growth rates of the juveniles obviously quickened. Growth rates of shell height were 136.4 μm/d (9-12 d), 307.3 μm/d (13-16 d) and 445.8 μm/d (17-20 d), respectively. It is suggested that the competent larvae of R. venosa can be induced to settle and metamorphose by the fresh seawater from the precipitation pond. Spats of C. gigas and S. broughtonii could be animal feeds of the juveniles of R. venosa.

    • Effects of dietary carbohydrate-to-lipid ratios on growth, blood biochemical indices, key enzymes activity and gene expression of hepatic glucose metabolism of juvenile GIFT Oreochromis niloticus

      2016, 40(9):1376-1386. DOI: 10.11964/jfc.20160410370

      Abstract (2333) HTML (521) PDF 779.88 K (1963) Comment (0) Favorites

      Abstract:The purpose of this study was to evaluate the effects of dietary carbohydrate-to-lipid (CHO:LIP) ratios on growth performance, plasma biochemical indexes, pyruvate kinase (PK) and phosphoenolpyruvate carboxykinase (PEPCK) activity and mRNA expression in liver of juvenile GIFT Oreochromis niloticus. Five isonitrogenous and isoenergetic diets were formulated to contain various CHO:LIP ratios ranging from 1.5 to 16.5. Each diet was tested in three replicates of fish[initial weight:(6.77±0.14) g] for 8 weeks in the recirculating aquacultural system. The results indicated that specific growth rate (SGR), weight gain rate (WGR), protein efficiency ratio (PER) increased significantly as dietary CHO:LIP varied from 3.9 to 7.0, while feed conversion ratio (FCR) showed the opposite variation trend. Crude lipid content of whole body decreaced as dietary CHO:LIP ratios increased, and was at the lowest level in 16.5 group, which was significantly decreased only compared with 1.5 group. Plasma concentrations of triglycerides and total cholesterol also significantly decreased in 16.5 group. However, serum glucose increased significantly as CHO:LIP ratios increased. High dietary carbohydrate significantly enhanced the activities and mRNA expression of liver PK, while the effects of high carbohydrate on the activity and mRNA level of liver PEPCK were in a contrary tendency. Based on the second-order polynomial regression analysis of SGR, the optimal dietary carbohydrate and lipid contents for fingerling GIFT O. niloticus were 49.69% and 9.53%, respectively, with a corresponding dietary CHO:LIP ratio of 8.4.

    • Stress response of grass carp (Ctenopharyngodon idella), Prussian carp (Carassius auratus gibelio) and black carp (Mylopharyngodon piceus) to handling

      2016, 40(9):1479-1485. DOI: 10.11964/jfc.20151110164

      Abstract (2117) HTML (555) PDF 1.10 M (1639) Comment (0) Favorites

      Abstract:The influence of handling stress on concentrations of cortisol, glucose and lactate in plasma, glycogen content in liver and activities of two enzymes (hexokinase and pyruvate kinase) responsible to glycolysis in liver of three cyprinidaes with different food habits, grass carp Ctenopharyngodon idella (herbivore), Prussian carp Carassius auratus prussianio (omnivore) and black carp Mylopharyngodon piceus (carnivore) was evaluated. The concentrations of cortisol, glucose and lactate in plasma of grass carp, Prussian carp and black carp significantly increased after handling. The content of liver glycogen of grass carp and black carp declined at 2 h after handling, while no significant difference was found in content of liver glycogen of Prussian carp before and after handling. The glucose concentration in plasma was higher in black carp than in grass carp and Prussian carp, while the content of liver glycogen was higher in Prussian carp than those in grass carp and black carp. A smaller increase in concentrations of lactate and glucose in plasma was observed in Prussian carp, suggesting that stress response induced by handling was lower in Prussian carp compared to grass carp and black carp. No significant difference was found in the activities of hexokinase and pyruvate kinase in liver of grass carp and Prussian carp before and after handling, while the activity of hexokinase in liver of black carp decreased at 2 h after handling. These results indicate that the increase of glucose concentration in plasma did not enhance the enzymes activities for glycolysis in liver of grass carp, Prussian carp and black carp.

    • Scanning electron microscopic observations of gill, digestive organ and crystal rod of Hyriopsis cumingii exposed to hypoxia and toxic Microcystis aeruginosa

      2016, 40(9):1486-1496. DOI: 10.11964/jfc.20150409846

      Abstract (2147) HTML (543) PDF 27.11 M (1229) Comment (0) Favorites

      Abstract:In order to clarify the toxic effects of hypoxia and toxic Microcystis aeruginosa on freshwater pearl mussel (Hygriopsis cumingii) and histopathology, scanning electron microscopy was used to investigate the histopathological changes in the gill, stomach, intestine and crystal rod of the freshwater pearl mussel. The results showed that a large number of gill shedding and columnar cell necrosis were observed from the seventh day in the combined experimental group with hypoxia and toxic algae, as well as cilia loss from gastrointestinal cavity surface and epithelial cell necrosis rupture, and the crystal rod completely disappeared from the fifth day; from the seventh day, in hypoxia group and toxic algae group, a small amount of gill and digestive tract lesions were observed, and in hypoxia group, crystal rod completely disappeared from the seventh day, while in toxic algae group, the crystal rod persists throughout the experiment. After the stress was removed, in the three exposed groups, the crystal rods did not return to the normal levels as in the control group. Therefore, hypoxia and toxic Microcystis aeruginosa caused unrecoverable damages in gill and digestive system of H. cumingii, and the combined stress group was most severely affected, and hypoxia caused more severe effects than the toxic Microcystis aeruginosa. This study provides a histopathological reference for the physiological adaptation mechanisms in H. cumingii exposed to hypoxia and toxic algae, and a theoretical basis for the feasibility of H. cumingii as a bioremediation tool species for improving eutrophic waters.

    • Effects of fish oil under different stored conditions on growth performance and antioxidant indexes in juvenile grouper (Epinephelus coioides)

      2016, 40(9):1387-1397. DOI: 10.11964/jfc.20160310326

      Abstract (2153) HTML (509) PDF 31.70 M (1312) Comment (0) Favorites

      Abstract:In order to study the effects of fish oil under different stored conditions on growth performance and antioxidant indexes in juvenile grouper, Epinephelus coioides. The conditions which the fish oil was oxidized are as follows:stored temperature:4℃, ambient temperature(AT):(31.5±3.5)℃; stored time:45, 90,135 d; supplementations of antioxidant:30 mg/Kg(EQ), 300 mg/Kg(HEQ). According to the different treatment conditions of fish oil, there were 14 groups:TF+EQ[positive control (fresh oil+ EQ)], TF[negative control (fresh oil)], T4℃+45d+EQ, T4℃+45d+HEQ, T4℃+90d+EQ, T4℃+90d+HEQ, T4℃+135d+EQ, T4℃+135d+HEQ, TAT+45d+EQ, TAT+45d+HEQ, TAT+90d+EQ, TAT+90d+HEQ, TAT+135d+EQ, TAT+135d+HEQ. Preparation of nitrogen and other fat diets fed the Epinephelus coioides, which is from juvenile grouper (18±0.02) g to 56 days. There were no significant differences between TF+EQ and T4℃+45d+EQ, but T4℃+45d+EQ was significantly higher than TAT+135d+EQ in WGR. The FCR of T4℃+45d+EQ, T4℃+45d+HEQ, TAT+45d+EQ and TAT+45d+HEQ were relatively low, which had no significant differences with TF+EQ and TF but significantly lower than TAT+135d+EQ and TAT+135d+HEQ (P<0.05). The cell structures of TF+EQ, T4℃+45d+HEQ were clear and obvious and the sizes of these fat droplets in cells were normal, but much more greater fat droplets in TAT+135d+EQ and TAT+135d+HEQ. The CAT of T4℃+135d+EQ, TAT+135d+EQ and TAT+135d+HEQ were significantly higher than TF+EQ and TF. The SOD and GSH-Px were highest inTAT+135d+EQ, which were significantly higher than TF+EQ and TF. The MDA of TF+EQ, TF, T4℃+45d+EQ, T4℃+45d+HEQ, TAT+45d+EQ and TAT+45d+HEQ in liver had no significant differences, but were significantly lower than the other groups. We found the growth performance and antioxidant index in juvenile grouper of T4℃+45d+EQ, T4℃+45d+HEQ, TAT+45d+EQ and TAT+45d+HEQ were relatively good, T4℃+45d+EQ, T4℃+45d+HEQ were better than TAT+45d+EQ and TAT+45d+HEQ. That shows short term storage, adding antioxidants are advantageous to storage of fish oil, besides 4℃ is better than ambient temperature.

    • Effects of dietary vitamin D3 level on transcriptomic response in yellow catfish (Pelteobagrus fulvidraco)

      2016, 40(9):1289-1298. DOI: 10.11964/jfc.20151110165

      Abstract (2301) HTML (525) PDF 1.19 M (2126) Comment (0) Favorites

      Abstract:The study was carried out to investigate the effects on possible gene regulation in intestine and kidney of yellow catfish (Pelteobagrus fulvidraco) under the treatments of different dietary vitamin D3 levels (0(VD0), 1243(VD2), 22 700(VD20) IU/Kg) by RNA-seq technique. 83 265 unigenes were selected from the 305 568 982 raw reads. The average length was 845.38 nt and N50 was 1620 nt. 29 224 genes were annotated by using Blast software. According to the results of gene ontology functional enrichment analysis, significant differently expressed genes Were mainly distributed on the pathways like cellular process, metabolic pathways and biological regulation. The results of KEGG pathway enrichment analysis showed that 1532 genes were involved in metabolic pathway, in which the affected genes were the most. Dietary vitamin D treatment significantly affected the genes expression. When compared with the control group (VD0 vs VD2/VD0 vs VD20), 380 genes were up-and down-regulated in common, in which 266 genes were up-regulated and 114 genes were down——regulated. Among three levels (VD0 vs VD2 vs VD20), 21 genes were up-and down-regulated in common, including 4 genes up regulated and 17 down-regulated. The results indicated that different dietary vitamin D3 levels significantly affected the gene expressions in intestine and kidney of yellow catfish. The information of the differentially expressed genes provided evidences about the growth, metabolism and immunological response of yellow catfish.

    • Nutritional composition and edible safety analysis of two estuarine bivalve clams, Meretrix meretrix and Mactra veneriformis in Shuangtaizi Estuary

      2016, 40(9):1497-1504. DOI: 10.11964/jfc.20160610422

      Abstract (2002) HTML (545) PDF 499.61 K (1514) Comment (0) Favorites

      Abstract:In order to understand the nutrient characteristics of Meretrix meretrix and Mactra veneriformis, the general nutrients (crude protein, crude fat and crude ash), amino acids, as well as heavy metals (Cd, As, Cr, and Hg) in edible part of these two clams were investigated. The results showed that the crude protein content (58.24 g/100 g) and crude fat (5.93 g/100 g) in M. meretrix was similar to those in M. veneriformis (57.38 g/100 g, and 6.18 g/100 g). The polysaccharides content in M. meretrix is approximately three times as high as that in M. veneriformis. The crude ash contents in M. meretrix and M. veneriformis were 18.35 g/100 g and 25.39 g/100 g, respectively. In dry sample, the total amino acids content was up to 44.87 g/100 g in M. meretrix and 37.20 g/100 g in M. veneriformis. Meanwhile, contents of the essential amino acids and delicious amino acids in M. meretrix (18.39 g/100 g, 18.89 g/100 g) were higher than those in M. veneriformis (15.16 g/100 g, 15.58 g/100 g). The ratio of essential amino acids content to total amino acids was close in both buried clams and more than 40%. Among four heavy metals including Cd, As, Cr, and Hg, the Cd exhibited the highest content. The content of Cd was 5.40 mg/kg in M. meretrix and 6.3 mg/kg in M. veneriformis. It was found that the contents of Cr and Hg in M. meretrix and M. veneriformis were lower than the standard for the toxic substance in the non-environmental pollution aquatic products and other limit standards. Whereas, Cd and As contents in both estuarine buried clams were higher than the standard. Based on the PMTDI health risk evaluation model, the contents of heavy metals in both buried clams posed no health risk to humans. In conclusion, the results suggest that M. meretrix and M. veneriformis are both suitable for diet and delicious additives. Furthermore, M. meretrix is a more ideal protein source for humans given the composition and content of protein and amino acid.

    • Effects of compound diet and fresh frozen Ammodytes personatus on growth, antioxidant ability and lipid metabolism of hybrid grouper (Epinephelus fuscoguttatus ♀×Epinephelus lanceolatus ♂) juveniles

      2016, 40(9):1398-1407. DOI: 10.11964/jfc.20150910061

      Abstract (2836) HTML (518) PDF 558.90 K (1763) Comment (0) Favorites

      Abstract:This study was conducted to determine the effects of commercial compound diet and fresh frozen Ammodytes personatus on growth, antioxidant ability and lipid metabolism of hybrid grouper (Epinephelus fuscoguttatus♀×E. lanceolatus♂) juveniles. Ten thousand five hundred and forty-two groupers[initial average weight (191.47±3.70) g] were randomly divided into two groups with three replicates at a stock density of one thousand seven hundred and fifty-seven, and commercial compound diet and fresh frozen A.personatus were allocated to each of two groups of fish for a feeding period of 75 days. The results showed that the weight gain rate (WGR), specific growth rate (SGR), condition factor (CF),feed conversion ratio (FCR) and whole body fat content of hybrid grouper juveniles fed with fresh frozen trash fish group were significantly higher than those of fish fed with commercial compound diet (P<0.05), and the hepatosmatic index (HSI), viscerosomatic index (VSI) were significantly lower than those of commercial compound diet group (P<0.05). But the survival rate (SR) and protein efficiency ratio (PER) had no difference between fresh frozen trash fish group and commercial compound diet group (P>0.05). The contents of total cholesterol (CHO), triglyceride (TG), and low density lipoprotein cholesterol (LDL-C) in serum of commercial diet group were significantly decreased compared with fresh frozen A.personatus group (P<0.05). The activities of glutamic-pyruvic transaminase (GPT) and glutamic-oxaloacetic transaminase (GOT) in commercial compound diet were significantly higher than those of fresh frozen A. personatus group (P< 0.05). There was an obvious decrease in levels of superoxide dismutase (SOD), total antioxidant capacity (T-AOC), catalase enzyme (CAT) and fatty acid synthase (FAS) (P<0.05), and a significant increase in activity of lipoprotein lipase (LPL) was observed in commercial diet group (P<0.05), while the levels of malic dehydrogenase (MDH) and carnitine palmitoyltransferase-Ⅱ (CPT-Ⅱ) were not impacted by two diets. Under given conditions, the fresh frozen A.personatust is better than commercial compound diet, and commercial compound diet may cause an adverse effect on liver of hybrid grouper juveniles. Therefore, it is necessary to optimize and adjust the nutrient formula of hybrid grouper juveniles by referring to the quality nutrition characteristics of fresh frozen A.personatus.

    • Effects of dietary curcumin on growth performance, body composition and serum antioxidant enzyme activity in juvenile turbot (Scophthalmus maximus)

      2016, 40(9):1299-1308. DOI: 10.11964/jfc.20160410369

      Abstract (2342) HTML (662) PDF 675.57 K (1871) Comment (0) Favorites

      Abstract:A 77-d feeding trial was conducted to evaluate the effects of dietary curcumin on growth performance, body composition and serum antioxidant enzyme activity in junvenile turbot (Scophthalmus maximus)[initial body weight (5.12±0.04) g]. Four isonitrogenous and isoenergetic practical diets (50.66% protein, 17.36% lipid) were formulated to contain graded levels (0, 0.02%, 0.04% and 0.06% dry weight) of curcumin. Each diet was randomly assigned to triplicate groups of 35 fish and the fish were fed twice daily to apparent satiation. The results showed that survival rate (SR), specific growth rate (SGR), feed intake (FI), hepatosomatic index (HSI) and viserosomatic index (VSI) were not significantly affected by the dietary curcumin levels. There were no significant differences observed in whole body moisture content among all treatments. Lipid content in whole body significantly decreased, while lipid content in liver and muscle significantly increased with increasing dietary curcumin levels. Fish fed diet with 0.04% curcumin showed lower protein content in whole body than other groups. The activity of serum SOD increased with increasing dietary curcumin levels, and fish fed diet with 0.06% curcumin had significantly higher value than the control group. The activity of serum CAT was opposite to the trend of serum SOD activity, and the group fed diet with 0.06% curcumin had higher value than the control group. The serum MDA content and GSH content were significantly decreased with increasing dietary curcumin levels. In conclusion, the results from the present study indicated that 0.06% dietary curcumin improved the serum anti-oxidation ability of juvenile turbot, but did not affect the growth of fish.

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