Supplementation of Protein Hydrolysates to a Low‐fishmeal Diet Improves Growth and Health Status of Juvenile Olive Flounder,Paralichthys olivaceus

An 11‐wk feeding trial was conducted to evaluate three different protein hydrolysates as feed ingredients in high‐plant‐protein diets for juvenile olive flounder. Five experimental diets were fed to juvenile olive flounder to examine the effect of three different protein hydrolysates on growth performance, innate immunity, and disease resistance against bacterial infection. A basal fishmeal (FM)‐based diet was regarded as a high‐FM diet (HFM) and a diet containing soy protein concentrate (SPC) as a substitute for 50% FM protein was considered as a low‐FM diet (LFM). Three other diets contained three different sources of protein hydrolysates, including shrimp, tilapia, and krill hydrolysates (designated as SH, TH, and KH), replacing 12% of FM protein. All diets were formulated to be isonitrogenous and isocaloric. Triplicate groups of fish (15.1 ± 0.1 g) were handfed one of the diets to apparent satiation twice daily for 11 wk and subsequently challenged against Edwardsiella tarda. Growth performance and feed utilization of fish fed hydrolysate‐supplemented diets were significantly improved compared to those of fish fed the LFM diet. Dietary inclusion of the protein hydrolysates significantly enhanced apparent digestibility of dry matter and protein of the diets. In the proximal intestine, histological alterations were observed in the fish fed the LFM diet. The fish fed the hydrolysate diets showed significantly longer mucosal fold and enterocytes and greater number of goblet cells compared to fish fed the LFM diet. Respiratory burst activity was significantly higher in fish fed the TH and KH diets than fish fed the LFM diet. Significantly higher immunoglobulin levels were found in fish fed SH and KH diets compared to those of fish fed the LFM diet. Dietary inclusion of the protein hydrolysates in SPC‐based diets exhibited the highest lysozyme activity. Significantly higher superoxide activity was observed in groups of fish fed the KH diet. Fish offered the protein hydrolysates were more resistant to bacterial infection caused by E. tarda. The results of this study suggest that the tested protein hydrolysates can be used as potential dietary supplements to improve growth performance and health status of juvenile olive flounder when they were fed a LFM diet.     

Evaluation of shrimp or tilapia protein hydrolysate at graded dosages in low fish meal diet for olive flounder (Paralichthys olivaceus)

A study was carried out to examine and optimize the inclusion levels of shrimp hydrolysate (SH) or tilapia hydrolysate (TH) in low fishmeal diets for olive flounder (Paralichthys olivaceus). A fishmeal (FM)‐based diet was considered as a high FM (HFM) diet, and a diet containing soy protein concentrate (SPC) as a FM replacer at 50% substitution level was regarded as a low FM (LFM) diet. Six other experimental diets were prepared by dietary supplementation of SH or TH to LFM diet at different inclusion levels of 15 g/kg, 30 or 45 g/kg in the expense of FM (designated as SH‐1.5, SH‐3.0, SH‐4.5, TH‐1.5, TH‐3.0 and TH‐4.5, respectively). After 10 weeks of a feeding trial, growth performance and feed utilization efficiency of fish were significantly higher in fish fed HFM, SH‐3.0, SH‐4.5, TH‐1.5 and TH‐3.0 diets compared to those of fish fed LFM diet. Intestine diameter, villus height and goblet cell counts of fish were significantly increased by dietary inclusion of SH or TH into LFM diet. Dry matter and protein digestibility of diets were significantly improved by SH or TH incorporation. Innate immunity of fish was significantly enhanced by dietary SH or TH supplementation into LFM diet. Disease resistance of fish was significantly increased against Edwardsiella tarda by dietary inclusion of SH and TH at the highest inclusion level (45 g/kg). The optimum inclusion level of SH or TH in a SPC‐based LFM diet could be ~30 g/kg and 15–30 g/kg, for olive flounder.     

Evaluation of Antarctic krill (Euphausia superba) meal supplementation in diets for olive flounder (Paralichthys olivaceus)

The supplemental effect of Antarctic krill meal (KM) into a low fish meal (FM) diet was evaluated for olive flounder (Paralichthys olivaceus). A 56% FM‐based diet was regarded as a high FM inclusion diet (HFM), and a low‐FM diet (LFM) was prepared by replacing 50% FM from the HFM. Four other diets were prepared by supplementing 3%, 6%, 9% and 12% KM into the LFM diet gradually replacing soy protein concentrate and tankage meal (designated as KM3, KM6, KM9 and KM12 respectively). Quadruplicate groups of fish were fed one of the diets for 12 weeks. The growth performance and feed utilization efficiency were improved by the dietary KM supplementation. Digestibility of dietary protein and dry matter was increased by the KM3‐9 diets. Haematocrit and haemoglobin were increased by KM supplementation. The innate immunity and antioxidant capacity assessed by Ig, antiprotease, lysozyme, GPx and SOD and the condition factor of fish were significantly increased by KM3‐9 diets. Moreover, goblet cell counts, villi length and fillet yield of fish were significantly improved by all the KM‐containing diets (KM3‐12). A 25‐day‐long challenge test with the Edwardsiella tarda pathogen showed that the cumulative mortality was higher in fish fed the LFM diet than in fish fed the HFM or KM‐supplemented diets. The results indicate that dietary KM supplementation in a LFM diet can increase growth performance and feed utilization efficiency, diet digestibility, intestinal development and functions, innate immunity and disease resistance of olive flounder. The recommended level of KM inclusion in a LFM diet seems to be 6.6% according to quadratic regression analysis.     

Effects of Dietary Supplementation of Barodon on Growth Performance,Innate Immunity and Disease Resistance of Juvenile Olive Flounder,Paralichthys olivaceus,Against Streptococcus iniae

This study was conducted to evaluate the effects of dietary supplementation of Barodon, an anionic alkali mineral complex, on growth, feed utilization, humoral innate immunity and disease resistance of olive flounder. A basal experimental diet was used as a control and supplemented with 0.1, 0.2, 0.3, 0.4, or 0.5% Barodon. Triplicate groups of fish (26.4 ± 0.2 g) were fed one of the diets to apparent satiation twice daily for 10 wk. The growth performance was enhanced (P < 0.05) linearly and quadratically in fish fed diets containing Barodon compared with that in fish fed the control. Feed utilization was significantly improved by Barodon supplementation. Serum lysozyme and antiprotease activities were increased quadratically in Barodon fed groups. Also, significantly higher superoxide dismutase activity was found in Barodon‐fed fish. Dietary supplementation of 0.1–0.3% Barodon resulted in significant enhancement of fish disease resistance against Streptococcus iniae. The findings in this study indicate that dietary supplementation of Barodon can enhance growth, feed utilization, innate immunity, and disease resistance of olive flounder and that the optimum level seems to be 0.1% in diets.     

Effects of dietary supplementation of citrus by‐products fermented with a probiotic microbe on growth performance,innate immunity and disease resistance against Edwardsiella tarda in juvenile olive flounder,Paralichthys olivaceus (Temminck & Schlegel)

Two consecutive studies were conducted to evaluate the dietary supplementation of citrus by‐products (CB) fermented with probiotic bacteria on growth performance, feed utilization, innate immune responses and disease resistance of juvenile olive flounder. In Experiment I, five diets were formulated to contain 0% (control) or 3% four different CB fermented with Bacillus subtilis (BS), Enterococcus faecium (EF), Lactobacillus rhamnosus (LR) and L. plantarum (LP) (designated as CON, CBF‐BS, CBF‐EF, CBF‐LR and CBF‐LP, respectively). During 10 weeks of a feeding trial, growth performance and feed efficiency were not significantly different among all the fish groups. However, fish fed CBF containing diets had significantly higher survivals than the CON group. Disease resistance of fish against Edwardsiella tarda was increased by the fermentation of CB. In Experiment II, we chose the BS as a promising probiotic and formulated five diets to contain 0%, 2%, 4%, 6% and 8% CBF‐BS. Growth performance was not significantly affected by the CBF‐BS supplementation during 6 weeks of a feeding trial. Innate immunity of fish was significantly enhanced by CBF‐BS supplementation. Myeloperoxidase and lysozyme activities were increased in a dose‐dependent manner by dietary CBF‐BS inclusions. In a consecutive challenge test against E. tarda, an increased disease resistance was found by CBF‐BS supplementation. These studies indicate that the fermentation process of CB with probiotic has beneficial effects on innate immunity and thereby increases disease resistance of olive flounder against E. tarda. Bacillus subtilis can be used as a promising probiotic microbe for by‐product fermentation in fish feeds.     

Interactive effects of dietary cholesterol and protein sources on growth performance and cholesterol metabolism of Japanese flounder (Paralichthys olivaceus)

A feeding trial was conducted to compare the effects of supplemental cholesterol in fish meal (FM), fish protein concentrate (FPC), soy protein isolate (SPI) and soy protein concentrate (SPC)‐based diets on growth performance and plasma lipoprotein levels of Japanese flounder (Paralichthys olivaceus). Eight isonitrogenous and isocaloric diets including FM, FPC, SPI or SPC as sole protein source with or without supplementation with 10 g cholesterol kg^?1 diet were fed to juvenile fish for 8 weeks. Dietary cholesterol supplementation significantly increased the feed intake and specific growth rate in fish fed SPI‐based diets, but decreased those in fish fed FPC‐based diets. In addition, cholesterol supplementation significantly increased the level of cholesterol and ratio of low‐density lipoprotein cholesterol to high‐density lipoprotein cholesterol in plasma of fish fed fish protein‐based diets, whereas no effects were observed in fish fed soy protein‐based diets. The hepatic lipid content of fish fed FPC‐, SPI‐ or SPC‐based diets were significantly increased by supplemental cholesterol, but no influence was observed in fish fed FM‐based diets. These results suggested that dietary protein source modify the growth‐stimulating action of cholesterol; cholesterol supplementation may increase the arteriosclerotic lesion in fish fed fish protein‐based diets and the incidence of fatty liver in fish fed soy protein‐based diets.     

Feeding fermented soybean meal diet supplemented with taurine to yellowtail Seriola quinqueradiata affects growth performance and lipid digestion

Feeding experiment was conducted to determine whether fermentation and taurine supplementation of soybean meal (SBM) fed to yellowtail improve growth performance and lipid digestion. Six diets were formulated and designated as SBM, SBM supplemented with taurine (SBM + T), SBM fermented by Bacillus spp. (FSBM1), SBM fermented by Lactobacillus spp. (FSBM2), FSBM2 supplemented with taurine (FSBM2 + T), and fish meal (FM). Yellowtail fingerlings with an initial body weight of 42 g were stocked in 200‐L tanks. The fish were fed, each diet was fed twice daily in two separate tanks for 8 weeks. Results showed that lipid digestibility of the SBM diet was significantly lower than that of the FM diet, but SBM diet lipid digestibility was improved by taurine supplementation and fermentation. Lipase activity in anterior intestinal digesta, lipid contents in liver and muscle, and bile acid concentrations in gallbladder and anterior intestinal digesta were significantly lower in fish fed SBM diet than in fish fed FM diet; these parameters were not significantly different between fish fed SBM + T, FSBM2, FSBM2 + T diets and FM diet. Although taurine supplementation or fermentation improved lipid digestibility compared with FM diet, growth parameters were only improved in the fish fed FSBM2 + T diet, indicating that apart from a lack of taurine, SBM contains other factors responsible for inferior growth of yellowtail. These factors can be partially eliminated by Lactobacillus spp. fermentation.     

Effect of dietary fish protein hydrolysate on growth,feed utilization and IGF‐I levels of Japanese flounder (Paralichthys olivaceus)

This study was conducted to investigate the effect of fish protein hydrolysate on growth performance, insulin‐like growth factor I (IGF‐I) levels and the expression levels of liver IGF‐I mRNA in juvenile Japanese flounder (Paralichthys olivaceus). Fish hydrolysate was produced by enzymatic (alcalase and flavourzyme) treatment and size‐fractionated by ultrafiltration. The permeate after ultrafiltration (UF) and the non‐ultrafiltered fish hydrolysate were tested as feed ingredients using high plant protein diets. Fish meal was used in the control diet (FM). The feeding trial lasted for 60 days, and fish fed with 37 g kg^?1 UF showed the best growth, feed efficiency, digestibility and protein utilization. Plasma IGF‐I level was examined with radioimmunoassay, and the expression levels of liver IGF‐I mRNA were evaluated using real‐time PCR normalized against the 18S rRNA gene. Plasma IGF‐I levels were significantly increased by inclusion of fish protein hydrolysate. Liver IGF‐I mRNA expression was significantly higher in fish fed with 37 g kg^?1 UF diet than fish fed with control diet. The results indicated that small molecular weight compounds from fish protein hydrolysate showed a positive effect on growth and feed utilization in juvenile Japanese flounder. Dietary fish protein hydrolysate could improve plasma IGF‐I levels and liver IGF‐I mRNA expression in Japanese flounder.     

Effects of dietary fish oil replacement by microalgae raw materials on growth performance,body composition and fatty acid profile of juvenile olive flounder,Paralichthys olivaceus

Replacing dietary fish oil with DHA‐rich microalgae Schizochytrium sp. and EPA‐rich microalgae Nannochloropsis sp. for olive flounder (Paralichthys olivaceus) was examined. Three experimental isonitrogenous and isolipidic diets with lipid source provided by 50% fish oil (F50S50), 50% (M50F25S25) and 100% microalgae raw material (M100) respectively were compared with a soybean oil (S100) diet as control. Triplicate groups of olive flounder juveniles (16.5 ± 0.91 g) were fed the experimental diets, and a group was fed the control diets for 8 weeks in a recirculation system. Results showed feed efficiency and growth performance were not significantly changed when fish oil (FO) was totally substituted by soybean oil (SO) or microalgae raw material (MRM). The whole‐body composition, lipid content of liver and muscle, and lipid composition of plasma were not significantly influenced by the total substitution of FO by MRM. The polyunsaturated fatty acids (PUFA) content of muscle and liver declined in fish fed S100 diet, whereas it was not significantly reduced in fish fed M50F25S25 and M100 diets. The total substitution of FO by MRM not only maintained the levels of arachidonic acid, EPA or DHA but also increased n‐3/n‐6 ratio. In conclusion, MRM as the sole lipid source is sufficient to obtain good feed efficiency, growth performance and human health value in olive flounder juveniles.     

Effect of tributyrin supplementation in high‐soya bean meal diet on growth performance,body composition,intestine morphology and microbiota of juvenile yellow drum (Nibea albiflora)

Six equal‐protein and equal‐lipid diets were formulated: the fish meal (FM) diet, the soya bean meal (SBM) diet with 40% of FM protein replaced by SBM protein and tributyrin (TB) diets with 0.05% (TB0.05), 0.10% (TB0.1), 0.20% (TB0.2) and 0.40% (TB0.4) tributyrin supplemented in the SBM‐based diet. Each kind of diet was randomly fed to triplicate tanks with 20 fish per tank. Fish were fed apparent satiation twice daily for 56 days. No significant difference in weight gain rate (WGR) and feed efficiency rate (FER) was observed between fish fed the FM, TB0.1 and TB0.2 diets (p > .05). Muscle histidine and arginine proportion of fish fed TB0.1 diet was significantly higher than that of fish fed the SBM diet (p < .05). Intestine morphology results indicated that the supplementation of 0.1% tributyrin significantly improved the mucosal fold height, microvilli length and microvilli density when compared with those of fish fed the SBM diet (p < .05). The supplementation of dietary tributyrin suppressed the pro‐inflammatory gene expression, which may be due to the improvement of physical barrier and modification of microbial communities, such as Acinetobacter, Rhodocyclaceae, Brevundimonas, Sphingopyxis, Hydrogenophaga, Methyloversatilis and Devosia. In conclusion, dietary 0.1% tributyrin supplementation in high‐soya bean meal diet improved growth performance, flesh quality and intestinal morphology structure integrity of yellow drum.     

Supplemental iron and phosphorus increase dietary inclusion of cottonseed and soybean meal in olive flounder (Paralichthys olivaceus)

A long‐term feeding experiment was conducted to investigate the use of cottonseed and soybean meal (CS) with iron and phosphorus supplements in diets for olive flounder (Paralichthys olivaceus). Olive flounder with an initial average size of 28.5 ± 0.35 g (mean ± SD) were divided into 15 groups (three tanks per dietary treatment) and fed 480 g kg^?1 crude protein diets in which each of five isonitrogenous diets was formulated to contain different levels of cottonseed/soybean meal (1 : 1) to replace fish meal (FM) with iron and phosphorus supplementations. The five experimental diets were as follows: diet 1 (control), 0%CS; diet 2, 20%CS; diet 3, 30%CS; diet 4, 30%CS + Fe&P; and diet 5, 40%CS + Fe&P. After 26 weeks of feeding trial, no significant differences were observed in weight gain, feed utilization and survival among all the treatments. The total gossypol accumulation in liver of fish fed diets supplemented with iron was significantly lower than that of fish fed diets without supplementation of iron. The results indicate that the addition of iron in diets could prevent the absorption of free gossypol. The findings in this study suggest that dietary supplements of iron and phosphorus could increase the inclusion of cottonseed and soybean meal for FM replacement in diets for marine fish species.     

Effects of the Dietary Fermented Tuna By‐product Meal on Growth,Blood Parameters,Nonspecific Immune Response,and Disease Resistance in Juvenile Olive Flounder,Paralichthys olivaceus

This study evaluated the effects of dietary fermented tuna by‐product meal (FTBM) in juvenile olive flounder, Paralichthys olivaceus. Five diets were formulated to replace fishmeal (FM) with FTBM at 0% (FTBM₀), 12.5% (FTBM_12.5), 25.0% (FTBM₂₅), 37.5% (FTBM_37.5), or 50% (FTBM₅₀). After 8 wk, weight gain, specific growth rate, and feed efficiency of fish fed FTBM₀ and FTBM_12.5 diets were significantly higher than fish fed the other diets (P < 0.05). Also, mean cumulative survival rates (%) of fish fed the FTBM₀ and FTBM_12.5 diets were significantly higher than those fed FTBM₅₀ diet at Day 9 postchallenge with Edwardsiella tarda (P < 0.05). Protein efficiency ratio of fish fed FTBM₀ and FTBM_12.5 diets was significantly higher (P < 0.05) than fish fed diets FTBM_37.5 and FTBM₅₀. Broken‐line regression analysis of weight gain showed an optimal FM replacement level of 10.65% with FTBM. Therefore, the optimal dietary inclusion of FTBM in juvenile olive flounder diets could be greater than 10.65% but less than 12.5% without any adverse physiological effects on fish health.     

Effect of various sources of dietary additive on growth,body composition and serum chemistry of juvenile olive flounder (Paralichthys olivaceus)

Asia traditional herbal feed additives have often been shown to have beneficial effects on disease resistance, growth, feed conversion and protein conversion. There have been few studies on the use of these feed additives in olive flounder diets. The present study was designed to determine if there are benefits associated with the inclusion of any of these traditional herbal feed additives in olive flounder diets. One thousand two hundred sixty juvenile fish averaging 6.5 g were randomly distributed into 36, 180‐L flow‐through tanks. No additive was included into the control (Con) diet. The aqueous type of additives: 2.5% bamboo (BB), 0.25% Prunus mume (PM), 2.5% Artemisia capillaris (AC), 2.5% cinnamon (CC), 2.5% Cirtus junos (CJ), 0.25% Cudrania tricuspidata (CT) and 2.5% Ficus carica (FC), and the powder type of additives: 1% Houttuynia cordata (HC), 1% safflower (SF) and 1% sea tangle (ST) were included into the experimental diets. Finally, 0.1% emulsifying oil (EO) was included into the experimental diet. Feed efficiency ratio (FER) of fish fed the BB, CT and SF diets was higher than that of fish fed the FC and HC diets. Protein efficiency ratio (PER) of fish fed the BB and SF diets was higher than that of fish fed the AC, FC and HC diets. None of serum criteria of fish was affected by any of dietary additives. Dietary additives did not affect weight gain and serum chemistry of olive flounder, but 2.5% bamboo extract and 1% safflower powder was effective to improve FER and PER.     

Potential of indigenous Bacillus spp. as probiotic feed supplements in an extruded low‐fish‐meal diet for juvenile olive flounder,Paralichthys olivaceus

A 12‐week feeding trial was designed to assess the probiotic potential of indigenous Bacillus amyloliquefaciens and/or Bacillus subtilis singly or in combination with Bacillus licheniformis in an extruded feed for olive flounder (Paralichthys olivaceus) juveniles. A high fish meal (FM) diet (control) and a low‐FM diet containing an alternative protein blend (30% FM replacement, FM30) were formulated. Three other experimental diets were prepared by inclusion of B. amyloliquefaciens (BA), B. subtilis (BS), or a mixture of B. amyloliquefaciens, B. subtilis, and B. licheniformis (BASL) into FM30 diet, with a final concentration of 10⁶ CFU/g diet. Results indicated that the FM30 diet was well tolerated by flounder, and the overall performance was not affected by dietary treatments. Lysozyme activity and total immunoglobulin level were significantly reduced in flounders when fed with the FM30 diet compared with the BASL and BA diets, respectively. The Bacillus additives neither enriched the relative abundance of the corresponding Bacillus spp. in the relevant gut microbiota of olive flounder nor modulated the presumptive gene functions of the gut microbiome. Despite the absence of growth‐promoting effect, the tested probiotics could still be economically viable for use as immunostimulants in commercial flounder diets with partial FM replacement.     

Organic acids blend as dietary antibiotic replacer in marine fish olive flounder,Paralichthys olivaceus

The present study was conducted to evaluate the efficacy of organic acid blends as dietary antibiotic replacer in marine fish olive flounder, Paralichthys olivaceus. Fish averaging 3.5 ± 0.05 g (mean ± SD) were fed one of the four experimental diets: (1) without antibiotic or organic acid (Control/CON); (2) with antibiotic—50 mg oxytetracycline per kg body weight per day (OTC); (3) with organic acid blend A—4 g/kg diet (OA_A); and (4) with organic acid blend B—4 g/kg diet (OA_B), for 10 weeks. At the end of the experiment, total intestinal bacterial counts in fish‐fed OA_A, OA_B and OTC were significantly lower than that of fish‐fed CON diet (p < 0.05). Further, the group of fish‐fed organic acid blends (OA_A, OA_B) or antibiotic (OTC)‐supplemented diets exhibited lower intestinal Vibrio sp. counts compared with fish‐fed CON diet. Disease challenge test with bacteria Edwardsiella tarda showed significantly lower cumulative mortality rates for the group of fish‐fed OA_A, OA_B or OTC than that of fish‐fed CON diet (p < 0.05). There were no negative effects on the growth, serological characteristics and proximate composition among the group of fish‐fed different experimental diets. Therefore, the present experiment demonstrates that blends of organic acid could be a promising alternative to dietary antibiotics for the preventive and/or curative health management in marine fish olive flounder aquaculture.     

Effects of Dietary Inclusion of Chlorella vulgaris on Growth,Blood Biochemical Parameters,and Antioxidant Enzyme Activity in Olive Flounder,Paralichthys olivaceus

This study was carried out to investigate the effects of dietary inclusion of defatted Chlorella on growth performance, body composition, blood biochemistry, and antioxidant enzyme activity in olive flounder. Four isonitrogenous (51% crude protein) diets were formulated to contain 0 (control), 5, 10, or 15% Chlorella meal (CM) (designated as Con, CM5, CM10, and CM15, respectively) and fed to triplicate groups of fish (104.4 g) to apparent satiation twice daily for 8 wk. At the end of the feeding trial, significant enhancement (P < 0.05) in growth performance was obtained at over 10% CM compared to fish fed the control diet. No significant changes in dorsal muscle and liver proximate composition were found following CM administration. The groups fed CM‐containing diets revealed significantly lower plasma cholesterol concentration than those fed the control diet. Dietary CM affected antioxidant enzyme activity; significantly higher plasma catalase activity was found in fish fed ≥10% CM and total antioxidant capacity increased in CM5 and CM10 groups compared to the control. However, plasma glutathione peroxidase and superoxide dismutase activities were not significantly influenced by dietary CM. Also, significant enhancement in 1,1‐diphenyl‐2‐picryl‐hydrazyl radical scavenging activity was found in dorsal muscle of fish fed CM15 diet compared to the control. The findings in this study showed that dietary inclusion of 10–15% CM can enhance growth performance and affect antioxidant enzyme activity and lipid metabolism in olive flounder.     

Single or combined effects of fructo‐ and mannan oligosaccharide supplements and Bacillus clausii on the growth,feed utilization,body composition,digestive enzyme activity,innate immune response and lipid metabolism of the Japanese flounder Paralichthys olivaceus

In this study, we examined the effects of the following eight experimental diets, which varied in fructo oligosaccharides (FOS), mannan oligosaccharides (MOS) and Bacillus clausii concentrations, on the Japanese flounder: control diet (no FOS, MOS and B. clausii), diet F (5 g kg⁻¹ FOS), diet M (5 g kg⁻¹ MOS), diet FM (2.5 g kg⁻¹ FOS + 2.5 g kg⁻¹ MOS), diet B (10⁷ cells g⁻¹B. clausii), diet FB (5 g kg⁻¹ FOS + 10⁷ cells g⁻¹B. clausii), diet MB (5 g kg⁻¹ MOS + 10⁷ cells g⁻¹B. clausii) and diet FMB (2.5 g kg⁻¹ FOS + 2.5 g kg⁻¹ MOS + 10⁷ cells g⁻¹B. clausii). Japanese flounder, initially weighing an average of 21 g, were distributed into 24 net cages at a stocking density of 20 fish per cage. Each diet was hand‐fed to three groups of fish twice daily for 56 days. The weight gain rate (WGR) in fish fed diets B, MB and FMB were significantly higher than in fish fed the control diet, where the fish fed diet FMB had the highest WGR. Fish fed any of the diets, except diets F and B, exhibited better feed conversion ratio than those fed the control diet. Diets MB and FMB significantly elevated intestinal protease activity compared with the control diet, but only the diet FMB promoted amylase activity. Feeding diets FB and FMB increased body protein deposition; additionally, feeding diets B, MB and FMB significantly reduced body lipid deposition. Lysozyme (LSZ) activity was significantly higher in fish fed diets B, FB, MB and FMB than in fish fed the control diet. All diets, except diet M, decreased triglyceride (TG) levels compared to the control diet. Low‐density lipoprotein cholesterol levels in fish fed diets F, FB and FMB were significantly lower than in fish fed the control diet. Without exception, no diets affected feeding rate, condition factor, body moisture, ash contents, phagocytic activity of leucocytes or cholesterol or high‐density lipoprotein cholesterol levels. Our results suggest that diets supplemented with FOS, MOS and B. clausii improved growth performance and health benefits of the Japanese flounder more than other diets or the control diet.     

Dietary choline requirement of juvenile olive flounder (Paralichthys olivaceus)

An eight‐week study was conducted to determine the optimum dietary choline level in juvenile olive flounder, Paralichthys olivaceus. Seven diets were prepared to contain 0, 250, 500, 750, 1,000, 2,000 and 3,000 mg/kg diet. Juveniles (5.9 ± 0.03 g; 5.5 ± 0.4 cm; mean ± SD) were randomly distributed into 21 tanks (25 fish/tank) and fed one of the diets in triplicates. Survival rate of fish fed the diet containing the lowest choline level was significantly lower than those of fish fed the other diets (p < 0.05). Final body weight, weight gain, specific growth rate, feed efficiency and protein efficiency ratio significantly increased with increasing choline levels up to 1,000 mg/kg diet. Whole‐body protein and lipid contents increased in accordance with choline levels up to 750 mg/kg diet, beyond which they plateaued. Liver and muscle lipid contents elevated with increasing choline levels up to 2,000 mg/kg diet. Plasma cholesterol esters, triglycerides, cholesterol and total lipids were significantly influenced by the graded choline levels; however, responses of those indices were not identical. Broken‐line analyses of weight gain and liver choline concentrations responding to the graded choline levels revealed that choline requirements of the juvenile flounder could be between 847 and 1,047 mg/kg diet.     

Dietary Protein Hydrolysates from Animal By-Products: Digestibility and Enzymatic Activity for Dourado Salminus brasiliensis

Fishmeal (FM) is a high-cost and scarce feedstuff. Notwithstanding, the formulation and processing of diets for carnivorous fish strongly rely on FM as a protein source, given its palatability and biological value. Animal by-product hydrolysates are high-quality feedstuff and can substitute FM in aquafeeds. This study evaluated the digestibility of hydrolysates from tilapia residue (TR), tuna head (TH), swine liver (SL), and poultry liver (PL), and the profile of digestive enzymes in juvenile dourado, Salminus brasiliensis, a carnivorous Characin, fed diets containing graded levels of the hydrolysates. Inclusion of hydrolysates in diet formulations lowered the pH of feed but did not alter feed intake by fish. Higher apparent digestibility coefficients (ADCs) of diets were recorded for fish fed diets containing TR and SL, and lower ADCs were recorded for fish fed diets containing TH. Protease and lipase activities in the animal’s stomach were higher, especially for those fed with diets containing SL. Amylase activity was higher in pyloric caeca, whereas in the intestines, the higher activity was seen in fish fed control and TR diets. Animal by-product hydrolysates were highly digestible for dourado, and the enzymatic profile of fish was influenced by the nutrient contents of diets.     

Soy protein concentrate as a substitute for fish meal in diets for juvenile Acanthopagrus schlegelii: effects on growth,phosphorus discharge and digestive enzyme activity

The current high demand and cost of fish meal (FM) necessities the evaluation of alternative plant protein ingredients in diets of farmed marine fish. A 56‐day feeding trial was performed to study the effects of replacement of FM with soy protein concentrate (SPC) in diets of Acanthopagrus schlegelii. Diets were prepared at levels of 0%, 30%, 47.5%, 65%, 82.5% and 100% SPC, respectively, replacing FM. The results indicated no significant differences (p > .05) in % weight gain (WG) and specific growth rate of fish fed S30% to S82.5% diets compared with the control diet while, further inclusion at 100% significantly depressed growth performance. SPC inclusion and phytase supplementation significantly affected the phosphorous discharge (P‐load) showing lowest value (3.83 ± 0.53 g/kg WG) in S100% compared to control (14.79 ± 0.37 g/kg WG) and in fish fed S30% diet (13.24 ± 0.89 g/kg WG) (p < .05). The results of this study showed that FM could be substituted up to 82.5% by SPC in the diet of Acanthopagrus schlegelii fingerlings (5.53 ± 0.12 g) without any adverse effects. Phytase supplementation SPC based diets could be effective in reducing the phosphorus load in the aquatic culture environment.