The effects of two dietary synbiotics on growth performance, haematological parameters, and non-specific immune responses in Japanese eel (Anguilla japonica).

Feed additives have attracted increased attention in aquaculture due to their ability to modulate fish gut microbiota, resulting in improved fish growth and immunity. This study assessed two synbiotics' effects in Japanese eels: Bacillus subtilis with mannooligosaccharide (MOS) and Enterococcus faecium with fructooligosaccharide (FOS). Six diets, including a control (CON), oxytetracycline (OTC), and four synbiotic diets - B.subtilis at 1 × 106 and 107 CFU/g with 5 g/kg MOS (BS6MO and BS7MO) and E. faecium at 1 × 106 and 107 CFU/g with 5 g/kg FOS (EF6FO and EF7FO) - were fed to triplicate groups of 20 fish averaging 6.00 ± 0.07g for eight weeks. Fish fed the BSMOS diets showed significantly higher weight gain (WG), specific growth rate (SGR), and feed efficiency (FE) compared to the CON and OTC diets (P < 0.05), but not significantly different from those fed the EFFOS diets. Weight gain, SGR of fish fed EFFOS were not significantly different from those fed all other diets (P > 0.05). Fish fed the OTC diet showed a higher mean aspartate aminotransferase (AST) level, though the difference was not statistically significant. The myeloperoxidase (MPO) activity of fish fed the BS7MO diet was significantly higher than in all other diets, and the superoxide dismutase (SOD) activity of fish fed the BS7MO diet was also significantly higher than in the EF7FO diet. Overall, the BSMOS synbiotic diets were significantly more effective than the CON diet in enhancing fish survival against Vibrio anguillarum. Our findings suggest that synbiotics can be a preferable alternative to antibiotics in aquaculture.

Evaluation of dietary fermented tuna by-product meal as partial replacement for unprocessed tuna by-product meal in fishmeal-based diets for juvenile olive flounder Paralichthys olivaceus.

This study examined the effects of feeding fermented tuna by-product (FTBP) on the growth, non-specific immune response, liver and intestinal morphology, and disease resistance of olive flounder Paralichthys olivaceus. Olive flounders (n = 20; 2 g) were randomly assigned into four dietary groups in triplicates. Fish were fed four test diets (50% crude protein; 10% crude lipid) for 10 weeks in which unprocessed tuna by-product (TBP) meal was replaced (on a protein basis) with 0% (FTBP0 as the control diet), 15% (FTBP15), 30% (FTBP30), and 60% (FTBP60) of FTBP protein. Results showed that growth performance, blood parameters, and proximate composition were not influenced by FTBP inclusion. Non-specific immune parameters such as superoxide dismutase activity in the FTBP30 and FTBP60 groups were significantly higher than in the FTBP15 and the control groups, whereas lysozyme and myeloperoxidase activities were not different. Liver histopathology revealed normal architecture in groups fed FTBP0 and FTBP15 diets although mild alterations were noted in the FTBP30- and FTBP60-fed groups. Intestinal villi height and muscular thickness were not significantly altered with FTBP inclusion. Moreover, higher cumulative survival rate was observed in the FTBP60-fed group than fish fed with the other diets following the 13-day challenge with E. tarda. Together, these results demonstrate that fermented tuna by-product meal could be included at 60% in olive flounder diets.

Effects of partial replacement of dietary fish meal by bioprocessed plant protein concentrates on growth performance, hematology, nutrient digestibility and digestive enzyme activities in juvenile Pacific white shrimp, Litopenaeus vannamei.

BACKGROUND: Bioprocessing of plant feedstuff can be a novel approach for reducing the overwhelming dependence on fish meal in aquaculture. The objective of this study was to evaluate the performance of Pacific white shrimp Litopenaeus vannamei fed solid-state fermented protein concentrates in order to replace fish meal in the diet. RESULTS: In the first trial, a group of 15 shrimp (average 3.88 g) were randomly distributed into aquaria in triplicate according to the experimental diets. Ten isonitrogenous (400 g kg-1 CP) and isolipidic (90 g kg-1 CL) diets were formulated to contain high-protein fish meal (HFM) and low-protein fish meal (LFM), and four types of bioprocessed protein concentrates (BPCs) as a replacement of fish meal (BPC-A, -B, -C and -D) each at 30% and 50% FM replacement levels. BPC-A was a solid-state fermented mixture of soybean and corn gluten meals; BPC-B was pre-treated acid-hydrolyzed BPC-A; BPC-C and BPC-D were BPC-A + 2% shrimp soluble extract (SSE) and BPC-B + 2% SSE, respectively. After 8 weeks, shrimp fed the HFM, BPC-B, BPC-C and BPC-D diets showed significantly higher growth performance at 30% FM replacement than those of shrimp fed the BPC diets at 50% FM replacement. Interestingly, shrimp fed the BPC-D diet could replace up to 50% FM replacement. In the second trial, the results show that apparent digestibility coefficients of feeds and apparent digestibility coefficients of ingredients for crude protein were significantly higher in fish fed the BPC-B, BPC-C and BPC-D diets. CONCLUSIONS: The results demonstrated successful partial replacement of high-protein fish meal using high-quality fermented protein concentrates from plant sources. © 2019 Society of Chemical Industry.

Heat-killed Bacillus sp. SJ-10 probiotic acts as a growth and humoral innate immunity response enhancer in olive flounder (Paralichthys olivaceus).

Investigations were carried out to evaluate and quantify the effects of dietary supplementation with heat-killed (HK) Bacillus sp. SJ-10 (BSJ-10) probiotic (1 × 108 CFU g-1) on the growth and immunity of olive flounder (Paralichthys olivaceus). Flounder (averagely 9.64 g) were divided into two groups, and fed control and HK BSJ-10 (HKBSJ-10)-inoculated diets for 8 weeks. Investigations were carried out on growth and feed utilizations, innate immunity, serum biochemical parameters, microvilli length, and pro- and anti-inflammatory cytokine gene (tumor necrosis factor [TNF]-α, interleukin [IL]-1ß, IL-6, and IL-10) transcriptions. Compared to control, HKBSJ-10 diet significantly (P < 0.05) enhanced weight gain and protein efficiency ratio, 1.17 and 1.11 folds respectively. Humoral innate immune parameters, lysozyme and superoxide dismutase in treatment group were also elevated by 1.34 and 1.16 folds. Similarly, an increased (P < 0.05) relative expressions of TNF-α, IL-1ß, IL-6 were recorded in liver (2.71, 3.38, and 4.12 folds respectively), and gill (2.08, 1.98, and 1.81 folds respectively) than that of controls. Moreover, after challenge with Streptococcus iniae (1 × 108 CFU mL-1), the HKBSJ-10-fed group exhibited significantly higher protection (P < 0.05) against streptococcosis compared to controls, validating the observed changes in immune parameters and induction on the cytokine-encoding genes. Therefore, HKBSJ-10 increases growth, modulates innate immune parameters, and protects olive flounders against streptococcosis.

Tuna byproducts as a fish-meal in tilapia aquaculture.

Potentiality of the use of tuna byproducts as a fish-meal replacement on Nile tilapia (Oreochromis niloticus) was examined for 84 weeks by tracking the concentrations of cadmium and mercury in the internal organs, muscles and fish whole body through generation including their immature eggs and their larvae. The results confirmed that the tuna byproducts can be used as a fish-meal substitute in tilapia aquaculture, because their acceptable ranges for cadmium and mercury consequently did not exceed the food safety values (both < 0.5 mg kg-1), despite their proportional increases in the fish body. The use of tuna byproducts as a protein source is expected to reduce the cost of feed with other fishmeal substitutes in tilapia aquaculture. However, fish (flounder) indiscriminately consuming tuna byproduct feed were prohibited and recalls of sales were issued by the government (July 2018, Republic of Korea), as the threshold for mercury in the fish bodies had been exceeded (0.6-0.8 mg kg-1). Further study of the use of tuna byproducts as fishmeal replacements for other species in aquaculture is needed, as concentration ratios can vary depending on the species.

Synergistic effects of dietary supplementation of Bacillus subtilis WB60 and mannanoligosaccharide (MOS) on growth performance, immunity and disease resistance in Japanese eel, Anguilla japonica.

This study evaluated the synergistic effects of dietary Bacillus subtilis WB60 and mannanoligosaccharide (MOS) in juvenile Japanese eel, Anguilla japonica. Seven treatment diets were formulated to contain three different levels of B. subtilis (0.0, 0.5, and 1.0 × 107 CFU/g diet denoted as BS0, BS0.5, and BS1, respectively) with two MOS levels (0 and 5 g/kg diet denoted as M0 and M5, respectively), and one diet with oxytetracycline (OTC) at 5 g/kg diet. Each diet (BS0M0 (CON), BS0M5, BS0.5M0, BS0.5M5, BS1M0, BS1M5, and OTC) was fed to triplicate groups of 20 fish averaging 9.00 ±â€¯0.11 g (mean ±â€¯SD) for eight weeks. Average weight gain, feed efficiency, specific growth rate and protein efficiency ratio of fish fed the BS0.5M5 and BS1M5 diets were significantly higher than those of fish fed CON, BS0.5M0 and OTC diets (P < 0.05). Significant increases in the nonspecific enzymatic activities (e.g., lysozyme and myeloperoxidase) were detected from fish fed the BS0.5M5, BS1M5, and OTC diets compared to the CON, BS0.5M0, and BS0M5 diets (P < 0.05). Whereas, immunoglobulin M expressions were recorded significantly higher for fish fed the BS0.5M5 and BS1M5 diets compared to those of fish fed the other diets (P < 0.05). Also, heat shock protein 70 mRNA levels of fish fed BS0.5M5 and BS1M5 diets were significantly higher than those of fish fed the CON diet (P < 0.05). Histological observations of the intestinal morphology showed healthier gut for fish fed BS0.5M5 and BS1M5 diets than those fed CON, BS0M5, and OTC diets. Additionally, resistance to bacterial challenge with Vibrio anguillarum was recorded significantly lower for fish fed the CON diet than those fed other diets (P > 0.05). Therefore, the results for growth performance, non-specific immune responses, intestinal morphology, and disease resistance demonstrated that supplementation of B. subtilis at 0.5 × 107 CFU/g diet and mannanoligosaccharide at 5 g/kg diet could have beneficial synergistic effects in Japanese eel. The isolated probiotic from eel and the selected prebiotic could lead to the development of a specific and potential synbiotic in Japanese eel aquaculture.

Synergistic effects of dietary Bacillus sp. SJ-10 plus ß-glucooligosaccharides as a synbiotic on growth performance, innate immunity and streptococcosis resistance in olive flounder (Paralichthys olivaceus).

Bacillus sp. SJ-10 (BSJ-10) was identified from traditional Korean fermented fish, the previously recognized prebiotic ß-glucooligosaccharides (BGO), and their combination as a synbiotic were prepared to evaluate their individual and synergistic effects in olive flounder (Paralichthys olivaceus). Four diets (one control and three treatments) were formulated containing neither BSJ-10 nor BGO (control), 1 × 108 CFU g -1 BSJ-10 (BSJ-10), 0.1% BGO (BGO), and 1 × 108 CFU g-1 BSJ-10 + 0.1% BGO (BSJ-10 + BGO). Triplicates of 15 fish (weight 10 ± 0.25 g) were randomly allocated to the four diet groups and fed one of the diets for 8 weeks. At the end of the experiment, fish weight gain (WG), specific growth rate (SGR), feed conversion ratio, and protein efficiency ratio in BSJ-10, BGO and BSJ-10 + BGO diets were positively modulated (P < 0.05) compared with control. Specially, WG and SGR were significantly (P < 0.05) higher in BSJ-10 + BGO than that of BSJ-10 and BGO (individual component). The innate immune parameters such as respiratory burst, superoxide dismutase, and lysozyme activity (LSZ) of fish fed BSJ-10 and BSJ-10 + BGO (both groups) were significantly (P < 0.05) higher than the control. Moreover, myeloperoxidase activity (MPO) and LSZ of fish fed BSJ-10 + BGO were significantly higher compared with individual component. Compared with control, intestinal BSJ-10 content, expression of interleukin (IL)-1ß in liver and kidney, and tumor necrosis factor (TNF)-α in liver were higher in both groups, but microvillus length was increased (P < 0.05) only in BSJ-10 + BGO. During in vivo challenge experiment with Streptococcus iniae (1 × 108 CFU ml-1), survival rate of fish was significantly higher in all treatment groups versus control. Moreover, in BSJ-10 + BGO, protection against S. iniae infection and transcription of TNF-α and IL-6 in gill were significantly (P < 0.05) higher than the individual component. Collectively, an improved WG, SGR, MPO, LSZ, transcription of IL-6 and TNF-α, and cumulative survival rate against streptococcosis clearly demonstrates a synergistic outcome of diet BSJ-10 + BGO as synbiotic in olive flounder.

Comparative evaluation of dietary probiotics Bacillus subtilis WB60 and Lactobacillus plantarum KCTC3928 on the growth performance, immunological parameters, gut morphology and disease resistance in Japanese eel, Anguilla japonica.

The current experiment was conducted to evaluate and compare the efficacy of two different probiotics Bacillus subtilis WB60 and Lactobacillus plantarum KCTC3928 in diet of Japanese eel, Anguilla japonica. Seven experimental diets were formulated to contain no probiotics (CON), three graded levels of B. subtilis at 106 (BS1), 107 (BS2), 108 (BS3) and L. plantarum at 106 (LP1), 107 (LP2), 108 (LP3) CFU/g diet. Twenty fish averaging 8.29 ± 0.06 g were distributed in to 21 aquaria and were randomly assigned to one of the experimental diets in triplicate groups. Average weight gain (WG), feed efficiency (FE), and protein efficiency ratio (PER) of fish fed B. subtilis at 107 (BS2) and 108 (BS3) CFU/g diet were significantly higher than those of fish fed other experimental diets (P < 0.05). Nonspecific enzymatic activities including lysozyme, superoxide dismutase (SOD), myeloperoxidase (MPO) from fish fed B. subtilis at 107 (BS2) and 108 (BS3) CFU/g diet were detected to be significantly higher than that from fish fed CON diet (P < 0.05). Whereas, level of intestine glyceraldehyde-3-phosphate dehydrogenase (GAPDH), heat shock protein 70, 90 (HSP70, 90) and immunoglobulin (IgM) from fish fed B. subtilis at 107 and 108 CFU/g diet were recorded to be significantly higher than those of fish fed other experimental diets (P < 0.05). Parameters for intestinal morphology and histology suggested a healthier gut for the fish fed108 (BS3) CFU/g diet as compared to other treatment groups. Whereas, results from the disease challenge test with bacteria Vibrio angulillarum showed significantly lower survival rate for fish fed CON diet than those of fish fed other experimental diets. Therefore, these results indicated that oral supplement of B. subtilis at 108 (BS3) CFU/g diet could be a more effective source of probiotic compared to L. plantarum in Japanese eel.

Comparison of the effects of dietary single and multi-probiotics on growth, non-specific immune responses and disease resistance in starry flounder, Platichthys stellatus.

An 8-week feeding trial was conducted to evaluate the effects of dietary probiotics on growth performance and non-specific immune responses in starry flounder, Platichthys stellatus. Fish averaging 46.5 ± 0.65 g (mean ± SD) were fed one of the six experimental diets; one control (Cont), and five other diets were prepared by supplementing single-probiotics 1 (Bacillus subtilis; SP1, 2 × 109 CFU kg-1 diet), single-probiotics 2 (Bacillus licheniformis; SP2, 2 × 109 CFU kg-1 diet), multi-probiotics 1 (Bacillus subtilis + Bacillus licheniformis; MP1, 2 × 109 CFU kg-1 diet), multi-probiotics 2 (commercial probiotics; Bacillus subtills + Bacillus licheniformis + Paenibacillus polymyxa + Aspergillus oryzae + Saccharomyces cerevisiae; MP2, 2 × 109 CFU kg-1 diet) and oxytetracycline (OTC) at 5 g OTC kg-1 diet. At the end of 8 weeks feeding trial, weight gain (WG) and specific growth rate (SGR) of fish fed SP1, MP1 and MP2 diets were significantly higher than those of fish fed control diet (P < 0.05). Superoxide dismutase (SOD) activity of fish fed MP2 diet was significantly higher than those of fish fed OTC diet (P < 0.05). Nitro blue tetrazolium (NBT) activity and lysozyme activity of fish fed SP1, MP1 and MP2 diets were significantly higher than those of fish fed OTC diet (P < 0.05). However, there was no significant difference among fish fed SP1, SP2, MP1 and MP2 diets. During the Edwardsiella tarda challenge test, the first mortality occurred on day 2. After the 14 days challenge test, cumulative survival rate of fish fed MP1 and MP2 diets were significantly higher than those of fish fed control diet (P < 0.05). However, there was no significant difference among fish fed SP1, SP2, MP1, MP2 and OTC diets in survival rate at the termination of the challenge test. Although there was little advantage in immunological parameters with fish fed MP diets, single and multi-probiotics were equally effective statistically. These results demonstrated that single or multi-probiotics had equal beneficial effect as an antibiotic replacer in terms of growth performance, non-specific immune responses and disease resistance in starry flounder.

Evaluation of Optimum Dietary Threonine Requirement by Plasma Free Threonine and Ammonia Concentrations in Surgically Modified Rainbow Trout, Oncorhynchus mykiss.

This study was carried out to evaluate the dietary threonine requirement by measuring the plasma free threonine and ammonia concentrations in rainbow trout, Oncorhynchus mykiss after dorsal aorta cannulation. A total of 70 fish (average initial weight 506±8.2 g) were randomly distributed into each of the 14 net cages (5 fish/cage). After 48 hours (h) of feed deprivation, each group was intubated at 1% body weight with one of the seven L-amino acid based diets containing graded levels of threonine (0.42%, 0.72%, 0.92%, 1.12%, 1.32%, 1.52%, or 1.82% of diet, dry matter basis). Blood samples were taken at 0, 5, and 24 h after intubation. Post-prandial plasma free threonine concentrations (PPthr) of fish 5 h after intubation with diets containing 1.32% or more threonine were significantly higher than those of fish intubated with diets containing 1.12% or less threonine (p<0.05). Post-absorptive free threonine concentrations (PAthr) after 24 h of intubation of the fish with diets containing 0.92% or more threonine were significantly higher than those of fish intubated with diets containing 0.72% or less threonine. Post-prandial plasma ammonia concentrations (PPA, 5 h after intubation) were not significantly different among fish intubated with diets containing 1.12% or less threonine, except the PPA of fish intubated with diet containing 0.42% threonine. Broken-line model analyses of PPthr, PAthr, and PPA indicated that the dietary threonine requirement of rainbow trout should be between 0.95% (2.71) and 1.07% (3.06) of diet (% of dietary protein on a dry matter basis).

The Optimum Feeding Frequency in Growing Korean Rockfish (Sebastes schlegeli) Rearing at the Temperature of 15°C and 19°C.

Two feeding trials were conducted to determine the optimum feeding frequency in growing Korean rockfish, (Sebastes schlegeli) reared at the temperatures of 15°C and 19°C. Fish averaging 92.2±0.7 g (mean±standard deviation [SD]) at 15.0±0.5°C and 100.2±0.4 g (mean±SD) at 19.0±0.5°C water temperature were randomly distributed into each of 15 indoor tanks containing 250-L sea water from a semi-recirculation system. A total of five feeding frequency groups were set up in three replicates as follows: one meal in a day at 08:00 hour, two meals a day at 08:00 and 17:00 hours, three meals a day at 08:00, 14:00, and 20:00 hours, four meals a day at 08:00, 12:00, 16:00, and 20:00 hours, and one meal every 2 days at 08:00 hour. Fish were fed at the rate of 1.2% body weight (BW)/d at 15°C and 1.5% BW/d at 19°C. At the end of 8 wks of feeding trial weight gain and specific growth rate were significantly higher at the fish fed groups of one meal a day and two meals a day at 15°C and fish fed groups of 1 meal every 2 days at 19°C were significantly lower than those of all other fish fed groups. Glutamic oxaloacetic transaminase and glutamic pyruvic transaminase of fish fed group at 1 meal every 2 days was significantly higher than those of all other fish fed groups in both experiments. Weight gain, specific growth rate and condition factor were gradually decreased as the feeding frequency increased. The results indicate that growing Korean rockfish 92 and 100 g perform better at 15°C than 19°C water temperature. As we expected, current results have indicated that a feeding frequency of 1 meal a day is optimal for the improvement of weight gain in growing Korean rockfish grown from 92 g to 133 g at 15°C and 100 g to 132 g at 19°C water temperature.

Evaluation of the Efficacy of Fermented By-product of Mushroom, Pleurotus ostreatus, as a Fish Meal Replacer in Juvenile Amur Catfish, Silurus asotus: Effects on Growth, Serological Characteristics and Immune Responses.

The present experiment was conducted to evaluate the efficacy of dietary fermented by-product of mushroom, Pleurotus ostreatus, (FBPM) as a fish meal (FM) replacer in juvenile Amur catfish, Silurus asotus. A total number of 225 fish averaging 5.7±0.1 g (mean±standard deviation) were fed one of the five experimental diets formulated to replace FM with FBPM at 0%, 5%, 10%, 20%, and 30% (FBPM0, FBPM5, FBPM10, FBPM20, and FBPM30, respectively). At the end of eight weeks of the experiment, average weight gain (WG) of fish fed FBPM0 or FBPM5 were significantly higher than those of fish fed FBPM20 or FBPM30 diets (p<0.05). However, there was no significant differences in WG among the fish fed FBPM0, FBPM5 or FBPM10, and between fish fed FBPM10 or FBPM20, and also between those fed FBPM20 or FBPM30 diets. Lysozyme activity of fish fed FBPM0 or FBPM5 were significantly higher than those of fish fed FBPM10, FBPM20 or FBPM30 diets (p<0.05). The chemiluminescent response of fish fed FBPM5 was significantly higher than those of fish fed FBPM0, FBPM20 or FBPM30 diets (p<0.05). Broken line regression analysis of WG suggested that the maximal dietary inclusion level for FBPM as a FM replacer could be 6.3% without any adverse effects on whole body composition and on serological characteristics. Therefore, these results may indicate that the maximal dietary inclusion level of FBPM as a FM replacer could be 6.3% in juvenile Amur catfish.

Effects of Feeding Rate and Water Temperature on Growth and Body Composition of Juvenile Korean Rockfish, Sebastes schlegeli (Hilgendorf 1880).

Three feeding trials were conducted to evaluate the effects of feeding rate and water temperature on growth and body composition of juvenile Korean rockfish, Sebastes schlegeli rearing at 3 different water temperatures. A total of 270 fish (each experiment) individual body weight (BW) averaging 16±0.3 g (mean±SD) were fed a commercial diet for 4 wk at 16°C, 20°C, and 24°C. At each temperature, triplicate tanks were assigned to one of 6 feeding rates: 1.5%, 2.5%, 2.8%, 3.1%, 3.4%, and satiation (3.7% BW/d) at 16°C, 1.9%, 2.9%, 3.2%, 3.5%, 3.8% and satiation (4.1% BW/d) at 20°C and 1.7%, 2.7%, 3.0%, 3.3%, 3.6%, and satiation (3.9% BW/d) at 24°C water temperature. Weight gains of fish in satiation and 3.4% groups at 16°C, in satiation and 3.8% groups at 20°C and in satiation and 3.6% groups at 24°C were significantly higher than those of fish in the other treatments (p<0.05). A broken line regression analysis of weight gain indicated that optimum feeding rates of juvenile Korean rockfish were 3.41% at 16°C, 3.75% at 20°C and 3.34% at 24°C water temperature. Results of the present study indicate that the optimum feeding rate could be >3.1% but <3.41% at 16°C, >3.5% but <3.75% at 20°C and >3.0% but <3.34% at 24°C. As we expected results suggest that fish performed better at 20°C than 16°C or 24°C water temperature and the optimum feeding rate could be 3.1% BW/d to 3.7% BW/d in 16 g of juvenile Korean rockfish.

Effects of Different Dietary Cadmium Levels on Growth and Tissue Cadmium Content in Juvenile Parrotfish, Oplegnathus fasciatus.

This feeding trial was carried out to evaluate the effects of different dietary cadmium levels on growth and tissue cadmium content in juvenile parrotfish, Oplegnathus fasciatus, using cadmium chloride (CdCl2) as the cadmium source. Fifteen fish averaging 5.5±0.06 g (mean±SD) were randomly distributed into each of twenty one rectangular fiber tanks of 30 L capacity. Each tank was then randomly assigned to one of three replicates of seven diets containing 0.30 (C0), 21.0 (C21), 40.7 (C41), 83.5 (C83), 162 (C162), 1,387 (C1,387) and 2,743 (C2,743) mg cadmium/kg diet. At the end of sixteen weeks of feeding trial, weight gain (WG), specific growth rate (SGR) and feed efficiency (FE) of fish fed C21 were significantly higher than those of fish fed C83, C162, C1,387 and C2,743 (p<0.05). Weight gain, SGR and FE of fish fed C0, C21 and C41 were significantly higher than those of fish fed C162, C1,387 and C2,743. Protein efficiency ratio of fish fed C0, C21 and C41 were significantly higher than those of fish fed C1,387 and C2,743. Average survival of fish fed C0, C21, C41 and C162 were significantly higher than that of fish fed C2,743. Tissue cadmium concentrations increased with cadmium content of diets. Cadmium accumulated the most in liver, followed by gill and then muscle. Muscle, gill and liver cadmium concentrations of fish fed C0, C21, C41 and C83 were significantly lower than those of fish fed C162, C1,387 and C2,743. Based on the ANOVA results of growth performance and tissue cadmium concentrations the safe dietary cadmium level could be lower than 40.7 mg Cd/kg diet while the toxic level could be higher than 162 mg Cd/kg diet.

Effects of dietary squid oil on breeding performance and embryonic and larval development of butter catfish Ompok pabda.

The present study was conducted to determine the effects of squid oil on the endangered butter catfish, Ompok pabda to confer enhanced breeding with embryonic and larval development for promoting its aquaculture. A total of 360 fish were obtained from the Brahmaputra River, Mymensingh, Bangladesh, of which, 60 fish stocked in each tank having an initial weight and length of 16.35 ± 0.57 g and 15.25 ± 0.38 cm, respectively, in the cisterns of 1.22 × 2.44 × 1.25 m (total 6 cisterns) maintained at 90 cm water depth. During the experimental period, a constant physico-chemical conditions of water such as temperature, pH, and dissolved oxygen (DO) were 26.5 ± 2 °C, 7.4 ± 0.2, and 6.7 ± 0.5 ppm, respectively, were maintained in each cistern. As the source of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), dietary 1 % squid oil (SQO) was supplemented in the diet of the treated group (SQO diet) to compare while the control group offered with basal diet without SQO supplementation (CON diet). The fecundity, spawning, fertilization, hatching rate, and survival rate of fry in SQO group were significantly (P < 0.01) higher than in the CON group. Moreover, better early embryonic and larval development of fish was observed in the SQO group i.e. size of fertilized egg diameter, growth and early developmental stages of larvae but not significantly different from the control group. Collectively, the results of the present study showed that dietary SQO supplementation improved the breeding and reproductive performances of butter catfish. The findings of this study could assist to develop a nutrient-rich diet for the better broodstock development of butter catfish at the farm level which may ultimately reduce the mortality and poor survival of offspring of this commercially important catfish species.

Evaluation of Gamma-Aminobutyric Acid (GABA) as a Functional Feed Ingredient on Growth Performance, Immune Enhancement, and Disease Resistance in Olive Flounder (Paralichthys olivaceus) under High Stocking Density.

Gamma-aminobutyric acid (GABA) is a non-protein amino acid that is found in the brain and central nervous system of animals as an inhibitory neurotransmitter. It has been shown to have a variety of physiological functions, including stress reduction and immune enhancement. This study investigated the effects of dietary supplementation with GABA on growth, serum biochemistry, innate immunity, and disease resistance in juvenile olive flounders (Paralichthys olivaceus) challenged with Edwardsiella tarda under high-stocking density. A control diet and three experimental diets were prepared, with 150 mg/kg (GABA150), 200 mg/kg (GABA200), and 250 mg/kg (GABA250) of GABA added to each diet, respectively. Each experimental diet was fed to olive flounders in triplicate with an initial weight of 12.75 g ± 0.3 g in 40 L tanks at two stocking densities: normal density (20 fish/tank) and high density (40 fish/tank). After 8 weeks of the feeding trial, growth, feed utilization, whole-body proximate compositions, blood analyses, and non-specific immune responses were measured, and challenge tests were performed. There were no significant differences in the weight gain (WG) and specific growth rate (SGR) among fish fed the GABA-supplemented diets at the two stocking densities. However, the normal-density groups showed significantly higher WG and SGR than the high-density groups (p < 0.05). There was no significant difference in feed efficiency and protein efficiency ratio among all groups. Moreover, there was no significant difference in the whole-body proximate composition analysis (p > 0.05). There were no significant differences in cortisol levels in fish fed the GABA at both densities, but the high-density group showed a significantly higher cortisol than the low-density group. Blood GABA significantly increased in a dose-dependent manner regardless of the density groups (p < 0.05). Superoxide dismutase activity showed significantly higher levels than the control group, but there was no significant effect of the stocking densities in fish fed the GABA diets (p < 0.05). Myeloperoxidase activities in fish fed the GABA200 and GABA250 diets showed significantly higher levels at both of the stocking densities (p < 0.05). Lysozyme activity was significantly higher in the GABA150 group than in the CON, GABA200, and GABA250 groups (p < 0.05). After 15 days of challenge tests with Edwardsiella tarda, the cumulative survival rates of the GABA150, GABA200, and GABA250 groups were significantly higher than that of the CON group (p < 0.05). The results suggested that the optimal dietary GABA level for juvenile olive flounder culture is 150 mg/kg, regardless of rearing density, to enhance growth, immunity, and disease resistance.

Interactive Effect of Dietary Gamma-Aminobutyric Acid (GABA) and Water Temperature on Growth Performance, Blood Plasma Indices, Heat Shock Proteins and GABAergic Gene Expression in Juvenile Olive Flounder Paralichthys olivaceus.

Gamma-aminobutyric acid (GABA) is an important inhibitory neurotransmitter in the central nervous system of living organisms and has the ability to reduce the magnitude of stress in humans and animals. In this study, we evaluated the supplemental effects of GABA on normal and high water temperature based on growth, blood plasma composition as well as heat shock proteins and GABA-related gene expression in juvenile olive flounder. For this, a 2 × 2 factorial design of experiment was employed to investigate the dietary effects of GABA at 0 mg/kg of diet (GABA0 diet) and 200 mg/kg of diet (GABA200 diet) in water temperatures of 20 ± 1 °C (normal temperature) and 27 ± 1 °C (high temperature) for 28 days. A total of 180 fish with an average initial weight of 40.1 ± 0.4 g (mean ± SD) were distributed into 12 tanks, of which, each tank contained 15 fish based on the 4 dietary treatment groups in triplicate. At the end of the feeding trial, the results demonstrated that both temperature and GABA had significant effects on the growth performance of the fish. However, fish fed the GABA200 diet had a significantly higher final body weight, weight gain and specific growth rate as well as a significantly lower feed conversion ratio than the fish fed the GABA0 diet at the high water temperature. A significant interactive effect of water temperature and GABA was observed on the growth performance of olive flounder based on the two-way analysis of variance. The plasma GABA levels in fish were increased in a dose-dependent manner at normal or high water temperatures, whereas cortisol and glucose levels were decreased in fish fed GABA-supplemented diets under temperature stress. The GABA-related mRNA expression in the brains of the fish such as GABA type A receptor-associated protein (Gabarap), GABA type B receptor 1 (Gabbr1) and glutamate decarboxylase 1 (Gad1) were not significantly affected by GABA-supplemented diets under normal or temperature stressed conditions. On the other hand, the mRNA expression of heat shock proteins (hsp) in the livers of the fish, such as hsp70 and hsp90, were unchanged in fish fed the GABA diets compared to the control diet at the high water temperature. Collectively, the present study showed that dietary supplementation with GABA could enhance growth performance, and improve the feed utilization, plasma biochemical parameters and heat shock proteins and GABA-related gene expression under the stress of high water temperatures in juvenile olive flounder.

Evaluation of Yeast Hydrolysate in a Low-Fishmeal Diet for Whiteleg Shrimp (Litopenaeus vannamei).

An eight-week feeding trial was performed to evaluate the effects of yeast hydrolysate (YH) supplementation in a low-fishmeal diet on the growth, immune responses, intestinal histology and disease resistance of whiteleg shrimp (Litopenaeus vannamei). Five experimental diets were produced by supplementing YH at 0 (CON), 0.5 (YH0.5), 1 (YH1), 2 (YH2) and 4 (YH4) % to a basal diet containing 10% fishmeal and compared with a positive control with 25% fishmeal (FM25). Shrimp with an initial average weight of 0.43 ± 0.005 g (mean ± SD) were stocked in 18 tanks and fed the experimental diets (38% protein and 8% lipid) four times a day. Results showed that shrimp fed the FM25 diet exhibited significantly higher final body weight, weight gain, specific growth rate and protein efficiency ratio than those fed CON, YH0.5, YH1 and YH2 diets (p < 0.05). However, there were no significant differences between shrimp fed the YH4 and FM25 diets (p > 0.05). In addition, there were no significant differences in whole-body proximate composition, hemolymph biochemical parameters and non-specific immune responses among treatments. Intestinal villi length and muscular layer thickness of shrimp fed the YH4 and FM25 diets were significantly higher than the other groups. At the end of the bacterial (Vibrio parahaemolyticus) challenge test, shrimp fed YH4 and FM25 diets showed a significantly higher survival rate than those of shrimp fed CON, YH0.5 and YH1 (p < 0.05). These results suggest that supplementing 4% YH in diet containing 10% fishmeal could beneficially influence growth, intestinal morphology and disease resistance of whiteleg shrimp.

Effect of Four Functional Feed Additives on Growth, Serum Biochemistry, Antioxidant Capacity, Gene Expressions, Histomorphology, Digestive Enzyme Activities and Disease Resistance in Juvenile Olive Flounder, Paralichthys olivaceus.

An 8-week feeding trial was executed to evaluate the efficacy of four functional feed additives in replacing antibiotics in juvenile olive flounder, Paralichthys olivaceus, fed with a low-fish-meal diet. A basal diet without feed additives was used as a control (CON); other diets were formulated by supplementing 0.50% taurine (TW), 0.30% peptide (PT), 0.23% mineral water (MW), 0.35% yeast-extracted nucleotides (GRO), 0.35% GRO + 0.50% taurine (GROTW), 0.35% GRO + 0.30% peptide (GROPT) and 0.35% GRO + 0.23% mineral water (GROMW) into the basal diet; in addition, one diet was supplemented with oxytetracycline (OTC) at 0.5% as a positive control. Triplicate groups of 25 fish with an average weight of 5.15 ± 0.06 g (mean ± SD) were fed one of the nine experimental diets. At the end of the feeding trial, the weight gain, specific growth rate and protein efficiency ratio of fish fed the GRO, GROMW, GROPT and GROTW diets were significantly higher than those of fish fed the CON diet (p < 0.05). The feed efficiency of fish fed the GRO, GROMW, GROPT and GROTW diets was significantly higher than that of fish fed the TW and OTC diets. However, the survival, hepatosomatic index, viscerosomatic index and condition factor of fish, as well as their whole-body proximate composition, were not significantly affected by the experimental diets (p > 0.05). The serum glutamic pyruvic transaminase of fish fed the GROPT diet was significantly lower than that of fish fed the CON diet. However, glutamic oxaloacetic transaminase, glucose and total protein were not significantly affected by the experimental diets (p > 0.05). The serum superoxide dismutase activity of fish fed the PT, TW, GRO, GROMW, GROPT and GROTW diets was significantly higher than that of fish fed the CON diet. The lysozyme activity of fish fed the PT, GRO, GROMW, GROPT and GROTW diets was significantly higher than that of fish fed the CON and OTC diets. The myeloperoxidase activity of fish fed the TW, GRO, GROMW, GROPT and GROTW diets was significantly higher than that of fish fed the CON, PT and MW diets (p < 0.05). The flounder growth hormone gene expression of fish fed the TW, GRO, GROMW, GROPT, GROTW and OTC diets was significantly higher than that of fish fed the CON, PT and MW diets (p < 0.05). The interleukin 1ß and interleukin 10 gene expressions of fish fed the GRO, GROMW, GROPT and GROTW diets were significantly higher than those of fish fed the CON, PT, TW and MW diets (p < 0.05). Intestinal histology showed a significantly higher villi length for fish fed the GRO, GROMW, GROPT and GROTW diets compared to that of fish fed the CON diet (p < 0.05). Digestive enzyme activities such as trypsin activity were significantly higher in fish fed the GROMW, GROPT and GROTW diets than those in the rest of the diet groups (p < 0.05). Amylase activity in fish fed the MW, GRO, GROMW, GROPT, GROTW and OTC diets was significantly higher than that of fish fed the PT, TW and CON diets (p < 0.05). On the other hand, the lipase activity of fish fed the TW, GRO, GROMW, GROPT and GROTW diets was significantly higher than that of fish fed the CON, PT, MW and OTC diets (p < 0.05). The cumulative survival rate of fish fed the PT, GROTW, GROPT and GROMW diets was significantly higher than that of fish fed the CON, TW and MW diets after thirteen days of the challenge testing. Overall, the results demonstrate that the GRO, GROMW, GROPT and GROTW diets could be beneficial feed additives to replace antibiotics in juvenile olive flounder fed low-fish-meal diets.

Re-evaluation of the Optimum Dietary Vitamin C Requirement in Juvenile Eel, Anguilla japonica by Using L-ascorbyl-2-monophosphate.

This study was conducted to re-evaluate the dietary vitamin C requirement in juvenile eel, Anguilla japonica by using L-ascorbyl-2-monophosphate (AMP) as the vitamin C source. Five semi-purified experimental diets were formulated to contain 0 (AMP0), 30 (AMP24), 60 (AMP52), 120 (AMP108) and 1,200 (AMP1137) mg AMP kg-1 diet on a dry matter basis. Casein and defatted fish meal were used as the main protein sources in the semi-purified experimental diets. After a 4-week conditioning period, fish initially averaging 15±0.3 g (mean±SD) were randomly distributed to each aquarium as triplicate groups of 20 fish each. One of five experimental diets was fed on a DM basis to fish in three randomly selected aquaria, at a rate of 3% of total body weight, twice a day. At the end of the feeding trial, weight gain (WG) and specific growth rate (SGR) for fish fed AMP52 and AMP108 were significantly higher than those recorded for fish fed the control diet (p<0.05). Similarly, feed efficiency (FE) and protein efficiency ratio (PER) for fish fed AMP52 were significantly higher than those for fish fed the control diet (p<0.05). Broken-line regression analysis on the basis of WG, SGR, FE and PER showed dietary vitamin C requirements of juvenile eel to be 41.1, 41.2, 43.9 and 43.1 (mg kg(-1) diet), respectively. These results indicated that the dietary vitamin C requirement could range from 41.1 to 43.9 mg kg(-1) diet in juvenile eel when L-ascorbyl-2-monophosphate was used as the dietary source of vitamin C.