Effect of dietary lipids and vitamin E on growth performance,body composition,anti‐oxidative ability and resistance to Aeromonas hydrophila challenge of juvenile Chinese mitten crab Eriocheir sinensis

Vitamin E supplement is important in protecting lipid oxidation and enhancing immunity of aquatic animals. A 10‐week feeding trial was conducted to evaluate the effect of dietary lipid and vitamin E on juvenile Chinese mitten crab Eriocheir sinensis. The experimental diets included three levels of vitamin E (0, 100 and 300 mg kg^?1) and two levels of lipid (6% and 9%). The 9% lipid diet significantly enhanced the body lipid of crabs compared with the 6% lipid diet. The vitamin E concentration of hepatopancreas increased with the increase in vitamin E in the diets regardless of dietary lipid levels. The hepatopancreas fatty acids, especially polyunsaturated fatty acid and highly unsaturated fatty acid, were significantly enhanced by vitamin E supplement or 9% dietary lipid. Vitamin E supplement significantly increased the total antioxidant capacity, superoxide dismutase, lysozyme and phenoloxidase activities of crabs compared with those fed the diets without vitamin E supplement. The hepatopancreas malondialdehyde of crabs fed 100 mg vitamin E kg^?1 was significantly lower than those fed 0 or 300 mg vitamin E kg^?1. The phenoloxidase activity of crabs fed the 9% lipid diet was significantly higher than those fed 6% lipid, irrespective of vitamin E levels. Vitamin E supplementation increased the bacterial resistance of juvenile crab. This study indicates that dietary lipid and vitamin E supplement do not affect crab growth and survival, but vitatmin E supplement at 100 mg kg^?1 can enhance immunity and antioxidant capacity of crab fed 6% or 9% dietary lipids.     

豆油替代鱼油对中华绒螯蟹幼蟹生长、非特异性免疫和抗病力的影响

以酪蛋白和明胶为蛋白源配制5组等氮等能的实验饲料, 分别为全鱼油(F1)、25%(F2)、50%(F3)和75%(F4)豆油替代组和全豆油组(F5), 每组4个重复, 投喂初始体质量为(0.27±0.03) g的中华绒螯蟹(Eriocheir sinensis)幼蟹6周, 评价豆油替代鱼油的饲料对幼蟹生长、非特异性免疫力和抗病力的影响。结果发现, 各组幼蟹的存活率差异不显著(P>0.05); F4组幼蟹的增重率和特定生长率显著高于F1组(P<0.05), 但与其他各组差异不显著(P>0.05)。F4组幼蟹的血清酸性磷酸酶和碱性磷酸酶活性均显著高于其他各组(P<0.05), F5组最低。幼蟹的血清中超氧化物歧化酶活性以F4组最高, 显著高于F3组(P<0.05), F3和F4组均显著高于其他各组(P<0.05), 其他F1、F2和F5三组间差异不显著(P>0.05)。F1组幼蟹的血清丙二醛含量最高(P<0.05), 其余各组之间则差异不显著(P>0.05); F4组幼蟹的血清酚氧化酶活性最高, 而F5组最低(P<0.05); 各组幼蟹的溶菌酶差异不显著(P>0.05)。实验结束后, 采用嗜水气单胞菌(Aeromonas hydrophila)攻毒10 d, 发现F5组幼蟹最早出现死亡, 且该组幼蟹的累积死亡率显著高于F1和F4组(P<0.05)。结果提示, 采用豆油完全替代饲料中的鱼油对幼蟹的生长影响不显著, 但会降低其抵抗感染疾病的能力; 用75%豆油替代鱼油可在明显提高幼蟹免疫机能和抗病力的情况下获得良好的生长性能。

     

A combination of vitamins C and E alleviates oxidized fish oil‐induced hepatopancreatic injury in juvenile Chinese mitten crab Eriocheir sinensis

Dietary lipid oxidation is a lasting concern to aquafeed manufacturers and aquaculture farmers. Here, we determined if there was an association between oxidized fish oil (OFO) and hepatopancreatic injury in Chinese mitten crab, and evaluated the alleviating effects of a combination of vitamins C and E on hepatopancreatic injury. There were significant reductions in the body weight gain, hepatopancreatic index, and feed efficiency in crabs that received OFO. Diets with OFO also resulted in reduced crude fat contents in the hepatopancreas and muscle tissues, and alternations in the hepatopancreatic fatty acid compositions, especially a reduction in polyunsaturated fatty acids. Moreover, dietary OFO exposure reduced the antioxidant defense and non‐specific immune response, caused hepatopancreatic injury, and even apparent whitening. An ultrastructural examination of the whitened hepatopancreas revealed that hepatopancreatic cells were injured at varying degrees following OFO exposure. The combination of vitamins C and E could efficiently maintain a constant fatty acid profile in the hepatopancreas exposed to OFO, thus alleviating OFO‐induced hepatopancreatic injury. In addition, a biochemical analysis indicated that a combination of vitamins C and E not only improved the antioxidant status by mitigating the harmful effects of oxidized oil, but also modulated non‐specific immune responses in the crab that received OFO. Overall, OFO‐induced hepatopancreatic injury may be alleviated by a dietary combination of vitamins C and E, and the feeding of rancid forage fish or diets that contain highly oxidized oil is likely a cause of hepatopancreatic necrosis that is frequently observed in farmed crabs.     

Effect of poly‐β‐hydroxybutyrate on growth,enzyme activity and intestinal microbial community of Chinese mitten crab,Eriocheir sinensis (Milne‐Edwards) juveniles

Poly‐β‐hydroxybutyrate (PHB) is microbial carbon and energy storage polymer, which can be degraded into water‐soluble β‐hydroxybutyric acid in the gastrointestinal tract of aquatic animals. A 60‐day culture experiment was performed using Chinese mitten crab, Eriocheir sinensis (Milne‐Edwards) juveniles with an average initial body weight of 0.74 ± 0.06 g which were fed a diet supplemented with 0%, 0.5%, 1%, 3% or 5% PHB. A PHB dietary supplementation of 1% and 3% significantly improved the body weight gain, moulting frequency and concomitantly reduced 2nd–3rd moulting intervals of the crabs (P < 0.05). The dietary PHB level positively related to hepatopancreatic pepsin, trypsin and lipase activity (P < 0.05). Increasing the dietary PHB also improved total superoxide dismutase activity, but reduced alkaline phosphatase and acid phosphatase activity in the serum of hemolymph (P < 0.05). A 16S rRNA gene analysis by denaturing gradient gel electrophoresis indicated that PHB supplementation led to a significantly higher range‐weighted richness, diversity and evenness of the gut bacterial community when dosed at 3% in the feed. The beneficial effects of PHB are discussed in terms of immune defense, metabolism and gut microbiota of the crabs.     

Label‐free quantification proteomics reveals the effects of dietary fish oil and soybean oil on the immune response of Chinese mitten crab,Eriocheir sinensis

The replacement of fish oil (FO) in Eriocheir sinensis can significantly reduce the cost of E. sinensis cultivation, while several studies have indicated that replacing FO with soybean oil (SO) could significantly reduce the resistance of E. sinensis to disease. However, the molecular mechanisms underlying these effects remain poorly understood. In this study, crabs were fed two diets containing FO or SO, following which a label‐free quantification proteomic analysis was employed. And the activity of enzymes involved in the nonspecific immune response was also measured. Growth performance was undifferentiated between the crabs fed with FO and SO. A total of 519 proteins were identified, and 70 proteins were significantly altered between the crabs fed the two different diets. Five proteins related to the immune response were identified to be differently expressed. C‐type lectin, haemocyanin subunit 6 and cryptocyanin were significantly downregulated, while fatty acid‐binding protein and catalase were highly expressed in the crabs fed SO. The activities of acid phosphatase, alkaline phosphatase, superoxide dismutase and phenoloxidase were all significantly changed in crab fed with different diets. These findings will provide novel insight into the molecular mechanism regarding the replacement of FO on the immune response of E. sinensis and provide evidences for the relationship between nutrition and immunity in E. sinensis.     

Effects of dietary cholesterol levels on moulting performance,lipid accumulation,ecdysteroid concentration and immune enzymes activities of juvenile Chinese mitten crab Eriocheir sinensis

Effects of dietary cholesterol levels on moulting performance, lipid accumulation, ecdysteroid concentration and immune enzymes activities of juvenile Eriocheir sinensis were investigated. Crabs were fed with feeds contained various cholesterols of 1100, 2100 and 3200 mg kg^?1 in both paddy fields and laboratory experiments. In paddy fields trial, the crabs fed with diets contained 3200 mg kg^?1 cholesterol achieved higher growth rate than those fed with diets contained no supplemental cholesterol (1100 mg kg^?1). In laboratory trial, moulting frequencies of crabs fed with diets contained 3200 mg kg^?1 cholesterol were higher than those of crabs fed with diets contained no supplemental cholesterol from the 6th to 10th moult. Further laboratory experiment indicated that intermoults of crabs fed with diets contained 3200 mg kg^?1 cholesterol significantly shortened compared with crabs fed with the basal feeds (1100 mg kg^?1 cholesterol). In the intermoult, total lipid content, ecdysterone concentration and three immune enzymes in crabs were increased with the increment of dietary cholesterol levels between the 7th and the 8th moult in laboratory experiments. Taken together, dietary cholesterol not only enhanced moulting performances of growth, survival and moult frequency, but also enhanced total lipid storage, ecdysterone concentration and three immune enzymes activities in the intermoult period.     

Effect of dietary lipid source and vitamin E on growth,non‐specific immune response and resistance to Aeromonas hydrophila challenge of Chinese mitten crab Eriocheir sinensis

Six purified diets were formulated to contain three lipid sources, fish oil (FO), linseed oil (LO) and soybean oil (SO), at 6% diet lipid crossing two levels of vitamin E (100 and 300 mg α‐tocopheryl acetate/kg diet) for each lipid source (FO100, FO300, LO100, LO300, SO100, SO300). The juvenile Chinese mitten crab, Eriocheir sinensis, respectively, fed on these diets with four replicates for 6 weeks. The crab weight gain (WG) and specific growth rate (SGR) were significantly affected by dietary lipid sources. No difference was found between the crabs fed two levels of vitamin E, but the WG and SGR were numerically higher in crab fed 300 mg/kg vitamin E than those fed the other level of vitamin E. The lipid source and vitamin E level could affect fatty acid composition in the hepatopancreas. The contents of saturated fatty acids (SAFA) and n‐3HUFA were significantly higher in the crab‐fed fish oil. The highest contents of n‐6PUFA and n‐3PUFA were found in the crab‐fed soybean oil and linseed oil respectively. The contents of SAFA, n‐3HUFA and n‐3PUFA were higher in the 300 mg/kg vitamin E treatment. A lower malondialdehyde (MDA) content and higher phenoloxidase (PO) activity were observed in the crab fed 300 mg/kg vitamin E. The results of this study indicate that the Chinese mitten crab fed the diet with 6% fish oil and 300 mg/kg vitamin E showed better growth, antioxidant capacity and resistance to Aeromonas hydrophila.     

Effects of dietary folic acid on growth,antioxidant capacity,non‐specific immune response and disease resistance of juvenile Chinese mitten crab Eriocheir sinensis (Milne‐Edwards, 1853)

An 8‐week study was conducted to determine folic acid requirement and its effect on antioxidant capacity and immunity in juvenile Chinese mitten crab Eriocheir sinensis (Milne‐Edwards, 1853), followed by a challenge assay with the pathogen Aeromonas hydrophila for 2 weeks. Folic acid was added to a basal diet at seven levels (0, 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mg folic acid kg^?1 diet), and a diet free of folic acid and vitamin B₁₂ was also included as a control. Crabs were fed twice daily in 32 tanks with 7.76–8.17 mg oxygen L^?1, 25.0–31.0 °C and 7.5–8.3 pH. Growth and feed efficiency were significantly greater in crabs fed ≥2.0 mg folic acid kg^?1, but not significantly different between crabs fed diets >2.0 mg folic acid. The superoxide dismutase activity and glutathione S‐transferase activity were highest in crabs fed ≥2.0 mg folic acid kg^?1, followed by those fed 0.5 and 1.0 mg folic acid kg^?1, and the control diet. The malondialdehyde content was highest in crabs fed the control diet, followed by those fed 0 mg folic acid kg^?1, and the lowest value occurred in those fed ≥0.5 mg folic acid kg^?1. Phenoloxidase activity and total haemocytes were significantly higher in crabs fed ≥2.0 mg folic acid kg^?1 than other diets. Crabs fed 2.0 mg folic acid kg^?1 had the highest lysozyme, acid phosphatase and alkaline phosphatase activities but the lowest cumulative mortality. The optimum dietary folic acid requirement by E. sinensis was estimated at 2.29–2.90 mg kg^?1 diet.     

Identification of SNPs in the 5′‐flanking region and 3′‐UTR of the MIH gene and their association with precocity of the Chinese mitten crab Eriocheir sinensis

Moult‐inhibiting hormone (MIH), an important regulator of steroidogenesis, inhibits the synthesis of ecdysteroid in Y‐organ (YO) and plays a significant role in the regulation of moulting and post‐embryonic development of crustacean. Because unsuccessful moulting have been widely observed in precocious crabs, we investigated whether genetic variants in the 5′‐flanking region and 3′‐untranslated region (3′‐UTR) of the MIH gene are associated with precocity of the Chinese mitten crab. Thirty individual DNA samples were sequenced to search for SNPs in the 5′‐flanking region and 3′‐UTR of the MIH gene. Five SNPs (g.196 T>A, g.230 C>T, g.305 T>C, g.323 C>A and g.372 C>T) in the 5′‐flanking region and 6 SNPs (g.2677 C>T, g.2759 T>A, g.2807 T>C, g.3042 A>G, g.3088 T>G and g.3295 T>G) in the 3′‐UTR of the MIH gene were selected for the individual genotyping in a two‐stage association study. We found that a SNP g.3088 T>G in the 3′‐UTR of MIH gene was consistently associated with precocity of the Chinese mitten crab in stage 1 and stage 2, with a per‐allele OR (Odds Ratio) of 1.469 (95% CI: 1.169–1.844) after two stages combined (P = 0.001). However, no significant associations were observed between the other 10 SNPs and precocity of the Chinese mitten crab. To our best knowledge, this is the first association study between various SNP genotypes and phenotype attributes in Chinese mitten crab. Our findings suggest that the SNP g.3088 T>G may be a candidate marker for effective marker‐assisted selection to decrease the precocity of the Chinese mitten crab in future studies.     

The effect of poly‐β‐hydroxybutyrate on the performance of Chinese mitten crab (Eriocheir sinensis Milne‐Edwards) zoea larvae

Two experiments were conducted regarding the effect of poly‐β‐hydroxybutyrate (PHB) on the survival and development of Chinese mitten crab (Eriocheir sinensis Milne‐Edwards) larvae. Different PHB delivery approaches (particulate, enriched in filter feeding prey, or a combination of both) and feeding levels were applied to crab larvae from the zoea 2 (Z2) up to the megalopa stage. Bio‐encapsulation into rotifers and Artemia at a PHB dose of 100 mg L^?1 seemed an efficient approach to deliver PHB to the larvae and resulted in a significantly enhanced survival, development rate and osmotic tolerance. The results indicate that PHB should only be applied as a feed additive rather than as a food source, and that the availability of an adequate amount of nutrients seems to be of major importance for the PHB to induce these beneficial effects.     

Dietary effects of poly‐β‐hydroxybutyrate on the growth performance,digestive enzyme activity,body composition,mineral uptake and bacterial challenge of rainbow trout fry (Oncorhynchus mykiss)

This study investigated the effects of diet supplementation with poly‐β‐hydroxybutyrate (PHB) on growth performance, digestive enzyme activity, lipid metabolism, mineral uptake and bacterial challenge of the rainbow trout fry (initial weight: 111.3 ± 13.7 mg) during a 6‐week experimental period. In the experimental set‐up, the commercial diet of rainbow trout fry was replaced with 0.5%, 1% and 2% PHB. The results of our work showed that the replacing of diets with PHB in experimental treatments decreases the weight of rainbow trout fry during the first 2 weeks before significantly increasing final weight at the end of the 6‐week period. PHB also improved digestive enzyme activity in experimental treatments. The highest total protease, pepsin activity and pancreatic enzyme secretion were observed with the 0.5% PHB treatment. A higher concentration of Na and K was observed in the whole body of the fry fed on 1% and 2% PHB‐supplemented diets. Enhanced survival rates occurred in all groups of fry after bath exposure to Yersinia ruckeri compared to rates in those fed the control diet. Our results suggest that the diet supplemented with PHB may improve growth performance, digestive enzyme activity and the functioning of the immune system. These positive effects could be considered for new applications in aquaculture.     

Effect of dietary β‐1,3‐glucan on the immune response of Litopenaeus vannamei exposed to nitrite‐N

β‐1,3‐Glucan with 1.0 g kg^?1 was supplemented to a basal diet [control (C)] with different feeding schedules: permanently β‐1,3‐glucan diet (BG) and 14 days BG–14 days control diet (C + BG). Growth and immunological responses [respiratory burst (RB), superoxide dismutase (SOD), catalase (CAT), lysozyme and total protein] in haemolymph, hepatopancreas and muscle of Litopenaeus vannamei were recorded after 84‐day feeding and exposure to nitrite‐N (20 mg L^?1) for 120 h. β‐1,3‐Glucan administration did not affect growth performance. However, as compared with control, elevated CAT and lysozyme activities were seen in haemolymph of both BG groups, while significantly higher activities of SOD, lysozyme and RB in hepatopancreas, and higher activities of CAT and lysozyme in muscle were only seen in C + BG group. After nitrite‐N stress, significantly higher haemolymph protein, and hepatopancreas activities of lysozyme and RB were observed in both BG groups, but significantly higher activities of haemolymph SOD was only seen in C + BG group. The mortality in groups BG and C + BG was significantly lower than that in group C, but C + BG group showed a trend of higher nitrite‐N resistance compared with BG group. Considering dietary cost and immunostimulatory effects, the feeding schedule with 14 days BG–14 days control diet is more recommended.     

Comparative effect of Freund's adjuvant and Aloe vera L. gel on the expression of TNF‐α and IL‐1β in vaccinated Cyprinus carpio L. with Aeromonas hydrophila C. bacterin

The comparative effects of Freund's and Aloe vera gel as adjuvants on the expression of IL‐1β and TNF‐α genes were studied in vaccinated common carp (Cyprinus carpio) with Aeromonas hydrophila bacterin. Fishes were intraperitoneally immunized with A. hydrophila bacterin in combination with Aloe vera gel or Freund's and also without any adjuvant. At day 28 after immunization, all groups were challenged by lethal dose of A. hydrophila (10⁷ cells/fish). Changes in the expression of IL‐1β and TNF‐α genes were evaluated in anterior kidney before challenge and 12, 24, 72 and 7 days postchallenge using quantitative real‐time PCR. Higher expression levels of both genes were observed in all vaccinated groups compared with non‐immunized group. Fishes which received Aloe vera gel showed higher expression of IL‐1β and TNF‐α in relation to animals which vaccinated with or without Freund's adjuvant. We concluded that Aloe vera gel in compared with Freund's adjuvant had a more stimulatory effect on the expression of immune‐related genes in vaccinated common carp and it can use as a novel adjuvant in aquaculture.     

Effects of dietary supplementation of A3α‐peptidoglycan on the growth,immune response and defence of sea cucumber Apostichopus japonicus

To determine the effects of A3α‐peptidoglycan (A3α‐PG) extracted from Bifidobacterium sp. on the growth, immune response and disease resistance of sea cucumber Apostichopus japonicus, a 70‐day feeding trial was conducted in this study. A total of 216 sea cucumbers were fed with four practical diets prepared from a commercial feed with different contents (0, 1.5, 2.5 and 4.0 g kg^?1) of A3α‐PG. The specific growth rate (SGR), total coelomocyte count (TCC), phagocytotic activity and activities of four immunological enzymes in both cell‐free coelomic fluid (extracellular, EC) and coelomocyte lysate supernatant (intracellular, IC), including acid phosphatase (ACP), alkaline phosphatase (ALP), peroxidase (POD) and superoxide dismutase (SOD), were measured at the end of the feeding trial. Finally, the animals were administered a 16‐day Vibrio splendidus challenge via intraperitoneal injection to test the potency of A3α‐PG on disease resistance. Compared with the control (0 g kg^?1 A3α‐PG), a significant increase (P < 0.05) in SGR was observed in the groups fed with 1.5 and 2.5 g kg^?1 A3a‐PG. The TCC, ranging from 7.25 × 10⁶ to 1.05 × 10⁷ cells mL^?1, was not significantly affected (P > 0.05) by A3α‐PG,. Coelomocyte phagocytotic activities in all of the A3α‐PG‐supplemented groups were significantly activated (P < 0.05), but no significant difference (P > 0.05) was observed. Sea cucumbers fed with 1.5 and 2.5 g kg^?1 A3α‐PG exhibited significant activation (P < 0.05) of EC/IC‐ACP, EC/IC‐ALP, and EC/IC‐POD activities. A significant increase in EC‐SOD activities (P < 0.05) was exhibited by all groups with A3α‐PG supplementation. The challenge test showed that animals fed with diets containing 2.5 and 4.0 g kg^?1 A3α‐PG had significantly lower cumulative mortalities compared with the control 16 days after exposure. All of the results presented here show that A3α‐PG can positively enhance the growth, immune response and disease resistance of sea cucumber, suggesting that dietary supplementation of A3α‐PG has potential applications in the health management of economic species of sea cucumber.     

Dietary vitamin E could improve growth performance,lipid peroxidation and non‐specific immune responses for juvenile cobia (Rachycentron canadum)

An 8‐week feeding trial was conducted to establish the dietary vitamin E requirement of juvenile cobia. The basal diet was supplemented with 10, 20, 30, 40, 60, 120 mg vitamin E kg^?1 as all‐rac‐α‐tocopheryl acetate. The results indicated that fish fed the diets supplemented vitamin E had significantly higher specific growth rate, protein efficiency ratio, feed efficiency and survival rate than those fed the basal diet. It was further observed that vitamin E concentrations in liver increased significantly when the dietary vitamin E level increased from 13.2 to 124 mg kg^?1. Fish fed the basal diet had significantly higher thiobarbituric acid‐reactive substances concentrations in liver than those fed the diets supplemented vitamin E. Fish fed the diets supplemented with 45.7 and 61.2 mg kg^?1 vitamin E had significantly higher red blood cell and haemoglobin than those fed the basal diet, while fish fed the diets supplemented with 61.2 and 124 mg kg^?1 vitamin E had higher immunoglobulin concentration than those fish fed the basal diet. Lysozyme and superoxide dismutase were significantly influenced by the dietary vitamin E level. The dietary vitamin E requirement of juvenile cobia was established based on second‐order polynomial regression of weight gain and lysozyme to be 78 or 111 mg all‐rac‐α‐tocopheryl acetate kg^?1 diet, respectively.     

Biomarkers of bone development and oxidative stress in gilthead sea bream larvae fed microdiets with several levels of polar lipids and α‐tocopherol

Although dietary marine phospholipids are able to improve culture performance of marine fish larvae in a further extend than soybean lecithin, both types of phospholipids (PL) markedly increase oxidative risk. The inclusion of a fat‐soluble antioxidant such as the vitamin E α‐tocopherol could allow a better control of oxidative stress. The objective of this study was to determine the combined effect of graded levels of α‐tocopherol with different levels and sources of krill phospholipids (KPL) and soybean lecithin (SBL) on growth, survival, resistance to stress, oxidative status, bone metabolism‐related genes expression and biochemical composition of sea bream larvae. Sea bream larvae were completely weaned at 16 dph and fed for 30 days seven microdiets with three different levels of PL (0, 40 and 80 g kg^?1 diet) and two of α‐tocopherol 1500 and 3000 mg kg^?1 diet. Sea bream larvae fed diets without PL supplementation showed the lowest survival, growth and stress resistance, whereas increase in PL, particularly KPL, markedly promoted larval survival and growth. However, feeding SBL markedly increased TBARs and GPX gene expression increasing the peroxidation risk in the larvae. Besides, KPL inclusion improved incorporation of n‐3 HUFA and, particularly, EPA into larval tissues, these fatty acids being positively correlated with the expression of BMP‐4, RUNX 2, ALP, OC and OP genes and to bone mineralization for a given larval size class. The increase in dietary α‐tocopherol tends to improve growth in relation to the n‐3 HUFA levels in the diet, denoting the protective role of this vitamin against oxidation. Indeed, dietary α‐tocopherol decreased the oxidative stress in the larvae as denoted by the reduction in larval TBARs contents and gene expression of SOD and CAT, but not GPX. Thus, increase in dietary α‐tocopherol effectively prevented the formation of free radicals from HUFA, particularly EPA, but did not affect the incidence of bone anomalies or the expression of genes related to osteogenetic processes.     

Dietary soya allergen β‐conglycinin induces intestinal inflammatory reactions,serum‐specific antibody response and growth reduction in a carnivorous fish species,turbot Scophthalmus maximus L.

This study was conducted to investigate the effects of dietary β‐conglycinin on the growth performance, digestion, gut morphology and immune responses of juvenile turbot (Scophthalmus maximus L.). Four diets were formulated to contain 0%, 2%, 4% and 8% purified β‐conglycinin. Triplicate groups of 30 fish were fed to apparent satiation twice daily for 12 weeks. Fish fed 4% and 8% dietary β‐conglycinin showed significantly reduced specific growth rate, feed efficiency ratio, apparent digestibility coefficient of nutrients and whole‐body lipid contents, as well as a profound infiltration of mixed leucocytes in the lamina propria and a significant decrease in the absorptive surface of distal intestine. The expression of pro‐inflammatory cytokines, TNF‐α and IL‐1β, in the distal intestine was significantly upregulated by 4% dietary β‐conglycinin, whereas a significantly lower expression level of IgM and anti‐inflammatory cytokine TGF‐β1 was observed in fish fed 8% dietary β‐conglycinin. Serum lysozyme and alternative complement pathway activity were first significantly enhanced by 2% dietary β‐conglycinin and then rapidly declined by 4% and 8% dietary β‐conglycinin. Respiratory burst activity of head kidney macrophages and serum superoxide dismutase activity were significantly suppressed by 4% and 8% dietary β‐conglycinin. Dietary β‐conglycinin (2–8%) significantly increased the level of specific antibody against β‐conglycinin in serum. Collectively, these results suggested that higher levels of dietary β‐conglycinin (4–8%) induced a variety of non‐specific and specific immune responses and intestinal mucosal lesions in turbot, resulting in inferior feed utilization and poor growth performance.