Effects of short‐chain fructooligosaccharides on growth performance and hepatic intermediary metabolism in turbot (Scophthalmus maximus) reared at winter and summer temperatures

The effect of short‐chain fructooligosaccharides (scFOS) incorporation on growth, feed utilization, body composition, plasmatic metabolites and selected liver enzyme activities of turbot juveniles reared at winter (15 °C) and summer (20 °C) temperatures was studied. Four comparable diets were formulated to contain circa 50 : 50 fish meal and plant ingredients as protein sources. Experimental diets included increasing levels of scFOS (0, 5, 10 and 20 g kg^?1). Final weight was higher at 20 °C, but thermal growth unit, feed efficiency, nitrogen and energy retention were better at 15 °C. scFOS supplementation did not affect fish growth performance. Fish reared at 15 °C had higher liver glycogen, visceral and hepatosomatic indices. Liver lipids, plasma triglycerides, total lipids, cholesterol HDL and LDL were higher in turbot reared at 20 °C. Malic enzyme, fatty acid synthetase, alanine aminotransferase and glutamate dehydrogenase activities were higher in fish reared at 15 °C. Malic enzyme was lower in turbot fed with 20 g kg^?1 scFOS compared to control diet; however, fatty acid synthetase presented an increasing trend as dietary scFOS increased up to 10 g kg^?1. Glutamate dehydrogenase activity was higher in fish fed the control diet. Results seem to indicate no benefits of scFOS incorporation to diets on growth performance of turbot.     

Effect of a dietary phospholipid supplementation on growth and fatty acid composition of European sea bass (Dicentrarchus labrax L.) and turbot (Scophthalmus maximus L.) juveniles from weaning onwards

Two 40-day feeding trials using extruded diets were conducted to assess the effect of a dietary phospholipid (PL) supplementation on growth, survival and fatty acid composition of European sea bass (Dicentrarchus labrax) and turbot (Scophthalmus maximus) from weaning onwards. Two dietary treatments (FO and PL) were tested; both had an identical extruded basis (92.5% total diet weight) coated with a different lipid fraction (7.5% total diet weight). Diet PL contained 2% egg yolk PL (69% pure). In diet FO the PL was replaced by hydrogenated coconut oil. The isolipidic diets contained an equal amount of fish oil ethyl esters providing 1.6% (% diet dry weight) of n-3 highly unsaturated fatty acids (HUFA). A diet water stability test showed no effect of the PL supplementation on the leaching of the dietary fatty acids. In both fish species weight, but not survival, significantly increased as a result of PL supplementation. Weaning onto the experimental diets resulted in similar changes in the relative percent levels of fatty acids in both species. In general, the percentage of saturated fatty acids levelled off after a rapid increase, while monoenes increased after an initial decrease. Total n-3 polyunsaturated fatty acids (PUFA) decreased and total n-6 PUFA remained almost constant. The major effect of the dietary PL on fish fatty acid composition was a 50% increase in n-6 and n-3 HUFAs compared to the PL-free FO diet. The rise in n-6 HUFA may have reflected the higher moiety in the dietary PL. On the other hand this was not the case for the n-3 HUFA since they represented only low levels in the PL fraction (0.1%) compared to that provided by the ethyl esters (1.6%) suggesting a more efficient incorporation of the PL n-3 HUFA than of the ethyl ester n-3 HUFA. A second hypothesis is that the dietary PL may have favored the incorporation of the dietary ethyl ester n-3 HUFA.     

Cross‐effects of dietary probiotic supplementation and rearing temperature on growth performance,digestive enzyme activities,cumulative mortality and innate immune response in seabass (Dicentrarchus labrax)

Probiotic influence on fish immune response and digestive capacity is extensively discussed in aquaculture. In this experiment, a feeding trial was carried out for 100 days to evaluate the cross‐effects of probiotic supplementation and rearing temperature (17, 20 and 23°C) in juvenile seabass (Dicentrarchus labrax). The experimental diet was supplemented with a commercial probiotic blend (Biomin AquaStar Growout) at 3 g/kg diet (5.23 × 10⁸ CFU/kg diet), and tested against a non‐supplemented diet (control). Growth performance and innate immune responses were analysed at 70 and 100 days of feeding, whereas digestive enzyme activities were determined at 100 days of feeding. At the end of the feeding trial, fish were subjected to a handling stress and cumulative mortality was recorded. Digestive enzyme activities were influenced by temperature, with α‐amylase and lipase activities peaking at the higher temperature (23°C) and trypsin at the lower temperature (17°C). Immune parameters showed a significant temperature versus feeding duration effect, with complement system (ACH50) and peroxidase peaking at 70 and 100 days of feeding, respectively. Poststress cumulative mortality was higher at the lowest temperature (17°C), especially in fish fed the control diet. In conclusion, water temperature was the main variable affecting the studied parameters, whereas the dietary probiotic supplementation had influence on the chymotrypsin activity and survival rate in seabass reared at 17°C.     

Siberian sturgeon (Acipenser baerii) larvae fed Artemia nauplii enriched with poly‐β‐hydroxybutyrate (PHB): effect on growth performance,body composition,digestive enzymes,gut microbial community,gut histology and stress tests

The effects of feeding Artemia nauplii enriched with or without poly‐β‐hydroxybutyrate (PHB) and/or highly unsaturated fatty acids (HUFA) on Siberian sturgeon (Acipenser baerii) larvae were investigated. Feeding larvae with PHB‐enriched nauplii (PHB, PHB + HUFA) decreased the growth performance of the larvae. PHB affected the body composition by increasing the lipid content of the whole body and decreasing total saturated, monoenoic, n3, n6 and decosahexanoeic acid (DHA) in the larvae, indicating that the PHB addition affected lipid metabolism. A high activity of pepsin was observed in the digestive extracts of PHB treatments (PHB, PHB + HUFA), while PHB suppressed amylase activity in the intestine of the larvae. Based on molecular analysis, PHB changed the microbial community in the distal intestine of the larvae. The highest counts of goblet cells were observed in the HUFA‐containing treatments (HUFA, PHB + HUFA), indicating that HUFA addition may improve the mucosal barrier defence system. The overall quality of the larvae was evaluated by exposing them to different salinities and ammonia stress levels. PHB decreased survival rates in these challenges. Our results show that optimal PHB doses for bio‐encapsulation into Artemia remain to be determined for further application at the earliest larval stages of sturgeon.     

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.     

Effects of different dietary carbohydrate‐to‐lipid ratios on growth,plasma biochemical indexes,digestive and immune enzymes activities of juvenile orange‐spotted grouper Epinephelus coioides

An 8‐week feeding trial was conducted to investigate the effects of dietary carbohydrate‐to‐lipid (CHO:L) ratios on growth, liver and muscle glycogen content, haematological indices, and liver and intestinal enzyme activity of juvenile grouper (Epinephelus coioides). Five isonitrogenous (496.0 g/kg protein) and isoenergetic (21.6 KJ/g gross energy) diets with varying CHO:L ratios of 0.65 (D1), 1.31 (D2), 2.33 (D3), 4.24 (D4) and 8.51 (D5), respectively, were fed to triplicate groups of 20 fish (average 10.02 ± 0.1 g) for 8 weeks. Results showed that the weight gain rate, specific growth rate and protein efficiency ratio (PER) of juvenile grouper increased first and then decreased with the increase in CHO:L ratio, reaching a maximum value in the D4 (CHO:L = 4.24) diet. The trend for the feed conversion ratio was opposite to the PER. Along with the diet CHO:L ratios, the apparent digestibility coefficients (ADC) of crude lipid and energy for the juvenile groupers decreased gradually, while the ADC of dry matter, liver and muscle glycogen level increased gradually. Total protein, triglycerides and cholesterol in serum were all maximized in the D4 diet and glucose in the D5 (CHO:L = 8.51) diet. Digestive enzyme activity in the intestine was significantly affected by dietary CHO:L ratio. Liver hexokinase, phosphofructokinase and glucose‐6‐phosphate dehydrogenase activity increased significantly as CHO:L ratio increased. Liver lysozyme and acid phosphatase activity in the groupers fed the D3 (CHO:L = 2.33) diet was significantly higher than that of other diets. Liver fructose‐1,6‐bisphosphatase and alkaline phosphatase activity reached a maximum value in the D4 diet and was significantly higher than that in the D1 diet. Taking the above results together, it can be concluded that an optimal dietary CHO:L ratio of 2.33 is suitable for grouper culture concerning growth performance and health.     

Sodium alginate supplementation modulates gut microbiota,health parameters,growth performance and growth‐related gene expression in Malaysian Mahseer Tor tambroides

This study investigated the effects of sodium alginate supplementation on gut microbiota composition, health parameters, growth performances and growth‐related gene expression of Malaysian mahseer. Five test diets were formulated by supplementing 0%, 0.1%, 0.2%, 0.4% and 0.8% sodium alginate. Triplicate groups of juvenile Tor tambroides (2.19 ± 0.05 g) were stocked in 15 aquaria (20 individuals per aquarium) and fed at 3.0% body weight per day for 60 days. PCoA and UPGMA analysis showed that gut bacterial community were more convergence in higher sodium alginate‐supplemented diets. The percentage of Porphyromonadaceae, Bacteroides, Plesiomonas and Shewanella were substantially higher and Aeromonas, Entomoplasmatales and Prevotellaceae were drastically lower in higher sodium alginate (0.2%–0.8%) diets. Sodium alginate supplementation (≥0.2%) significantly improved the haematocrit value and respiratory burst activity of T. tambroides. Growth performances and feed utilization were significantly higher in 0.2%–0.4% sodium alginate‐supplemented diets. The increased growth rate of T. tambroides was governed by both hyperplastic and hypertrophic muscle growth. Real‐time PCR data demonstrated that most of the growth‐related genes were significantly upregulated in 0.2%–0.4% sodium alginate‐supplemented diet. Finally, it can be concluded that sodium alginate should be supplemented at 2 g/kg in practical fish feed formulation.     

Dietary supplement of Enterococcus faecalis on digestive enzyme activities,short‐chain fatty acid production,immune system response and disease resistance of Javanese carp (Puntius gonionotus,Bleeker 1850)

This study investigated the effects of Enterococcus faecalis on digestive enzyme activities and short‐chain fatty acid production in fish intestine, resistance against Aeromonas hydrophila and humoral immunity response by 3 experiments on Javanes carp (Puntius gonionotus). The experiment 1 revealed that diet supplemented with E. faecalis significantly (P < 0.05) increased protease and lipase activities compared to control fed fish. Moreover, E. faecalis supplementation significantly enhanced the production of propionic and butyric acid in the intestine, while no significant difference (P > 0.05) in acetic acid production was observed. In the challenge study (experiment 2), fish were injected (intraperitoneal) with 10⁷ A. hydrophila per ml and survival was significantly improved when fish were fed diet supplemented with E. faecalis compared to control fish. In experiment 3, dietary E. faecalis affected immune system response as fish fed the probiont and exposed to 10⁶ A. hydrophila per ml displayed significantly elevated antibody levels compared to control fed fish. Fish fed diet supplemented with E. faecalis but not exposed to the pathogen revealed significantly higher antibody level than control fish (P < 0.05). Therefore, E. faecalis can be used as a probiotic in Javanese carp farming.     

饲料中大豆黄酮对大菱鲆生长、消化酶活力、抗氧化力及肠道结构的影响

为研究大豆黄酮在大菱鲆幼鱼中的营养生理作用,本实验在以鱼粉为主要蛋白源的基础饲料中添加不同剂量的大豆黄酮(0、5、10、20和100 mg/kg)来配制5种等氮等脂的实验饲料,并通过12周的投喂养殖实验评估饲料中不同剂量的大豆黄酮对大菱鲆幼鱼生长性能、消化酶活力、抗氧化力以及肠道结构的影响。结果表明:与对照组相比,饲料中添加不同剂量的大豆黄酮对大菱鲆幼鱼的存活率(98.89%~100.00%)、终末体质量(21.24~24.42 g)、特定生长率(1.81~1.98%/d)、饲料效率(1.01~1.11)、摄食率(1.43~1.51%/d)及形体指标均没有产生显著性影响;饲料中添加大豆黄酮显著降低了大菱鲆幼鱼鱼体的粗蛋白(15.41%~15.59%)和粗脂肪(3.19%~3.93%)含量,但对鱼体的水分(77.41%~79.70%)和灰分(3.46%~3.81%)含量未产生显著性影响;饲料中添加10~100 mg/kg大豆黄酮显著提高了大菱鲆幼鱼的胰蛋白酶活力(35.26~40.66 U/g prot),但胃蛋白酶(31.75~49.56 U/mg prot)、肠蛋白酶(10.00~14.79U/mg prot)、胃淀粉酶(0.10~0.25 U/mg prot)和肠淀粉酶(0.05~0.17 U/mg prot)的活力在各处理组间没有显著性差异。饲料中添加5、10和20 mg/kg的大豆黄酮显著降低了血清丙二醛(10.67~11.17 nmol/mL)的含量,并显著提高了大菱鲆幼鱼血清超氧化物歧化酶(51.05~53.36U/mL)和谷胱甘肽过氧化物酶(551.40 U/mL)的活力;饲料中添加不同浓度的大豆黄酮对大菱鲆幼鱼后肠肠道结构完整性没有显著性影响,但10~20 mg/kg大豆黄酮显著促进了肠道组织结构的发育和成熟,提高了大菱鲆幼鱼后肠肠绒毛的高度(391.26~401.48μm)。研究表明,大豆黄酮(5~100 mg/kg)对大菱鲆的生长没有显著性的影响,但饲料中适量的大豆黄酮(10~20mg/kg)可以显著提高大菱鲆幼鱼的消化酶活力和抗氧化能力,并促进肠道绒毛的发育。     

Effects of five compound attractants in high plant‐based diets on feed intake and growth performance of juvenile turbot (Scophthalmus maximus L.)

An experiment was conducted to evaluate the effects of five attractants on feed intake, growth performance and appetite‐regulating genes expression for juvenile turbot (Scophthalmus maximus L.). Fish were fed with seven diets: fish‐meal‐based diet (FM), fermented soybean meal replacing 45% fish meal (FSBM) and other diets with 1% compound attractant A (CA‐A) consisting of amino acids, compound attractant B (CA‐B) consisting of quaternary ammonium bases, compound attractant C (CA‐C) consisting of nucleotides, compound attractant G (CA‐G) consisting of the combination of CA‐A, CA‐B and CA‐C and blue mussel protein hydrolysate (BMH) on the basis of FSBM diet respectively. Daily feed intake in CA‐A group was significantly higher, but that in CA‐B group was significantly lower than FSBM group. Similarly, specific growth rate was significantly enhanced in CA‐A and CA‐G groups and were significantly decreased in CA‐B group. Protein efficiency ratio and protein retention in CA‐B group were significantly lower than FSBM group. Furthermore, BMH supplement increased the mRNA expression of Ghrelin in the intestine of fish. These results showed that CA‐A and CA‐G supplement could enhance growth in plant‐based diets, but CA‐B supplement had the opposite effect, which might be associated with lower feed intake and lower feed utilization.     

Effect of dietary encapsulated organic salts (Na‐acetate,Na‐butyrate,Na‐lactate and Na‐propionate) on growth performance,haemolymph, antioxidant and digestive enzyme activities and gut microbiota of juvenile narrow clawed crayfish,Astacus leptodactylus leptodactylus Eschscholtz, 1823

A 63‐day experiment was done to study the effects of four levels (5, 10, 20 and 50 g/kg) of encapsulated organic salts (Na‐acetate, Na‐butyrate, Na‐lactate and Na‐propionate) on the growth indices and haemato‐immunological responses of crayfish Astacus leptodactylus leptodactylus (4.38 ± 0.08 g). Crayfish were distributed at 51 1,000‐L tanks (17 treatments at triplicate). The highest values of final weight (27.86 g), specific growth rate (2.94% body weight per day) and survival rate (96%) were observed in the crayfish fed the 20 g/kg of encapsulated Na‐propionate diet (p < .05). The highest activities of phenoloxidase (7.4 U/min), superoxide dismutase (7.80 U/min) and lysozyme (9.40 U/min) were observed in the gut of crayfish fed the 20 g/kg of encapsulated Na‐propionate diet (p < .05), as well as the highest activities of alkaline protease (10.70 U/mg), lipase (9.10 U/mg), amylase (9.60 U/mg) and the lactobacillus count (p < .05). Broken line regression model of SGR and phenoloxidase activity suggested that the optimum dietary levels of encapsulated Na‐acetate, Na‐butyrate, Na‐lactate and Na‐propionate could be 30.7, 31.8, 31.4 and 33.5 g/kg, respectively, in crayfish reared in culture conditions.     

Effects of dietary supplementation of probiotics on the growth,activities of digestive and non‐specific immune enzymes in hybrid grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀)

Effects of Bacillus cereus BC‐01, Lactobacillus acidophilus LAG01, Clostridium butyricum CBG01 and their combinations as supplementation on the growth performance, digestive enzyme activities and serum non‐specific immunity of hybrid grouper (Epinephelus lanceolatus ♂×E. fuscoguttatus ♀) were assessed. Seven different diets, that is one control diet (basal feed without any probiotics, CT) and six treatment diets containing single B. cereus (Bs), L. acidophilus (Ls) and C. butyricum (Cs) at 1.0 × 10⁹ cfu/kg feed, and also their combinations in equal proportion at 1.0, 2.0 and 3.0 × 10⁹ cfu/kg feed (BLC1, BLC2 and BLC3) were prepared respectively. After 60‐day feeding trial, the final weight, specific growth rate,food consumption, food conversion efficiency and apparent digestibility coefficient of fish in Ls and BLC3 were significantly higher compared with the control (p < .05). The activities of pepsin and trypsin in the intestine of fish for Ls and BLC3 were significantly higher relative to the control (p < .05). Relative to controls, significantly enhanced amylase and lipase activities in proximal intestine except for Cs and BLC1 and lipase activities in distal intestine except for Cs were observed (p < .05). Meanwhile, activities of superoxide dismutase in the serum of fish for all treatments, lysozyme and catalase in Ls and BLC3, and glutathione peroxidase except for Cs were significantly enhanced (p < .05). Based on the above, dietary supplementation of single L. acidophilus at 1.0 × 10⁹ cfu/kg or combination of three strains at 3.0 × 10⁹ cfu/kg for hybrid grouper is recommended.