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There is a considerable interest in supplementing fish diets with bio-functional substances such as polyunsaturated fatty acids (PUFAs), and research is being conducted to evaluate their effects on fish physiology, particularly on breeding performances. The objective of this study was to find out how squid-derived lipids (PUFAs) influence the Rohu carp, Labeo rohita, in terms of reproduction, and embryonic and larval growth. The experimental fish were provided with formulated diet for a duration of 90 days, containing 1 % squid-extracted lipids as a primary source of polyunsaturated fatty acids (PUFAs). To determine the accuracy of the formulation, a proximate analysis of the feed ingredients and the formulated feeds were conducted. The results demonstrated remarkable reproductive outcomes during the spawning trial. Observations in fish treated with PUFAs revealed a significant improvement (p

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

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.

Song-Gang® bio-stone (SGS) is a microporous crystalline hydrated aluminosilicate which has found various applications because of their very unique physiochemical characteristics such as ion exchange and absorptive-desorptive properties. Significant progress has been made in recent years on applications of these inorganic adsorbents in different industries including agriculture, aquaculture, water and wastewater treatment. This review article intends to summarize the published reports on the applications of SGS in aquaculture industry. SGS application as a feed additive to enhance fish growth and promote their health and nutritional parameters is the most important discussed areas. According to the technical data that are discussed in this review, SGS should be considered as a material with tremendous potential for application in the aquaculture industry. Considerable amounts of research works are under way to explore other opportunities for application of SGS to benefit aquaculture industry.

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 1β 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.

The red-pink pigments produced by Arthrobacter bussei reportedly possess antioxidant and antibiotic properties, in addition to serving as food colorants and providing nutritional benefits. Despite the industrial potential of A. bussei, research on this bacterium has remained limited since it was first identified. This study evaluated the effects of dietary supplementation with A. bussei isolated from the Saemangeum Reservoir on the growth performance, hematological parameters, antioxidant capacity, and innate immunity of Litopenaeus vannamei. A total of six experimental diets were prepared and evaluated through a feeding trial. A basal feed was used as the control (CON), and the fishmeal content of the experimental diets was partially replaced with A. bussei powder (0.2%, 0.4%, 0.6%, 0.8%, and 1.0%; AB-1, AB-2, AB-3, AB-4, and AB-5, respectively). Shrimp (mean ± standard deviation, 0.40 ± 0.01 g) were randomly stocked in 50-L acrylic tanks (18 tanks with 25 shrimp per tank). Experimental diets containing 38.5% of crude protein and 5.4% crude lipid was supplied to the shrimp four times daily (9:00, 13:00, 17:00, 21:00) at 6–14% of shrimp body weight for 6 weeks. The experimental groups (AB-2, AB-3, AB-4, and AB-5) fed with A. bussei powder showed significantly higher final body weight (FBW), weight gain (WG), feed efficiency (FE), and specific growth rate (SGR) than the CON group. The mean glutamyl oxaloacetic transaminase and glutamic-pyruvate transaminase levels of the experimental groups supplemented with A. bussei powder were significantly lower than those of the CON group. The experimental groups supplemented with A. bussei powder exhibited higher lysozyme, superoxide dismutase and glutathione peroxidase activity than the CON group. Our findings suggested that A. bussei powder can improve the growth rate, FE, health, antioxidant capacity, and non-specific immunity of L. vannamei. We recommend that the diets of Pacific white shrimp be supplemented with ≥ 0.4% A. bussei powder.

We investigated the nine experimental diets containing fish meal (FM) and/or fish meal analog (FMA) as the major source of animal protein to determine the optimum FMA level as the substitute of FM protein in the diet of juvenile Japanese eel. In addition, two natural feed additives such as Song-Gang stone (SG) and Yucca meal (YM) were supplemented in the diet to evaluate their efficacy as the immunostimulants. The diets are as follows: 100% FM + 0% FMA in diet (FMA0), 90% FM + 10% FMA in diet (FMA10), 80% FM + 20% FMA in diet (FMA20), 70% FM + 30% FMA in diet (FMA30), 60% FM + 40% FMA in diet (FMA40), FMA0 + 0.4% SG (FMA0SG), FMA0 + 0.1% YM (FMA0YM), FMA20 + 0.4% SG (FMA20SG), and FMA20 + 0.1% YM (FMA20YM). Nine groups of Japanese eel each with three replicates were distributed (initial weight of 9 ± 0.2 g) in rectangular tanks receiving flow through water. Each group of the treatment consisted with 15 fish and fed one of the diets for 8 weeks. At the end of the feeding trial, fish fed with the FMA0 and FMA10 diets showed no significant differences in weight gain (WG), specific growth rate (SGR), feed efficiency (FE), and protein efficiency ratio (PER). Meanwhile, fish fed with FMA20, FMA30, and FMA40 diets showed significantly lower WG, SGR, FE, and PER than the fish fed with the FMA0 (control) diet. In addition, there were no significant differences among fish fed with the SG- and YM-supplemented diet groups. However, lysozyme activities in fish fed with the FMA10, FMA20, FMA30, and FMA40 were significantly lower than the fish fed with the FMA0SG, FMA0YM, FMA20SG, and FMA20YM diets. After 7 days of injection with V. Anguillarum, cumulative survival rates of fish fed with the FMA0SG and FMA0YM diets were significantly higher than the FMA0 diet group. The results revealed that the FMA could replace up to 10% of FM as a protein source in the diet of Japanese eel and both of the natural feed additives (SG and YM) could improve replacing rates of FMA from 10% to 20% without compromising growth and health status of fish.

β-glucan is considered as an effective immunostimulant because of its binding capacity to different receptors on leukocytes leading to the stimulation of immune responses including bactericidal activity, cytokine productivity, and survival fit ability at cellular levels. In response to immune cell surface receptors, β-glucan stimulates to release cytokines and chemokines. It has been found that these signaling proteins eventually stimulate the immunocompetent cells in fish such as monocytes, macrophages, and neutrophils for killing pathogens by phagocytosis, oxidative burst, and cytotoxic killing activities. They also procreate immunological memories and specific antibodies through activation of T and B lymphocytes. Researchers have proved that β-glucan can modulate some important biochemical (serum hemoglobin, serum protein, and total hemocyte count) and immunological (lysozyme activity, phagocytic activity, oxidative burst activity, and phenoloxidase activity) properties providing more competent immune profile for treating fish and aquatic organisms. β-glucan-supplemented fish showed limited sensitivity of genes involved in acute inflammatory reactions. Findings have shown that β-glucan exerts a positive impact on fish and aquatic organisms’ immunity, enhancing their disease resistance by increasing functional and decreasing deleterious responses. This review focuses on the basic bump of β-glucan on fish and shellfish immunity and recent information on the uses of β-glucan in progressive aquaculture.

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.

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.

Abstract Five experimental diets were formulated to evaluate the effects of dietary enzymatically hydrolyzed tuna by-product on growth, non-specific immune responses, and hematology of juvenile rainbow trout (Oncorhynchus mykiss). A basal diet with 50% of fishmeal was used as control (CON) and four other diets replaced 12.5% (TBB12.5), 25% (TBB25), 37.5% (TBB37.5), and 50% (TBB50) of fish meal in the CON diet. Juvenile rainbow trout (4.87 ± 0.05 g) were randomly distributed into 15 tanks (50 L) and fed 3–4% of wet body weight two times a day. At the end of 7 weeks of feeding trial, weight gain, specific growth rate, feed efficiency, and protein efficiency ratio of fish fed CON diet were significantly higher than those of fish fed TB50 diet (P 0.05). There were no significant differences in GPT levels among fish fed CON, TBB12.5, TBB25, and TBB37.5 diets. Also, there were no significant differences in lysozyme, superoxide dismutase, glucose, and total protein levels in all experimental diet (P > 0.05). The broken-line analysis indicated that the minimum dietary level of enzymatically hydrolyzed tuna by-product to replace fishmeal could be 29.7% in rainbow trout. These results indicated that the optimum level of dietary enzymatically hydrolyzed tuna by-product could replace greater than 29.7% but less than 37.5% of fishmeal in juvenile rainbow trout diet.

Molluscs are the most important resources among all the seafood items in South-East Asian countries. However, very little information available on nutritional value of molluscs in these regions. In this study, we evaluated the 7 economically important species of molluscs in terms of proximate composition, amino acids profile, fatty acids profile, cholesterol and heavy metal contents in the bivalves (mussels, oysters, clams and cockles) and univalve (snail) collected from freshwater and marine environments of Bangladesh. The results of the proximate analyses revealed that significantly higher amount of crude protein contents were present in marine water oysters, clams and cockles (59.3 ± 0.3 to 75.4 ± 0.2%) than the freshwater mussels and snail (36.9 ± 0.4 to 49.6 ± 0.6%) on dry matter basis. However, carbohydrate contents were significantly higher in freshwater mussels and snail (30.2 ± 0.9 to 57.3 ± 0.2%) compared to the marine water bivalves (8.1 ± 0.4 to 20.2 ± 0.6%). Crude lipid contents were ranged from 2.5 ± 0.2 to 11.2 ± 0.1% and ash from 11.4 ± 0.1 to 16.8 ± 0.6% among the bivalves and snail species. The amino acid contents were comparatively higher in marine water bivalves than their freshwater counterparts. Saturated fatty acid contents were found to be higher in marine water bivalves than the freshwater mollusc species. The results also show that the omega-3 (eicosapentaenoic acid, EPA and docosahexaenoic acid, DHA) fatty acids were comparatively higher in oysters, clams and cockles in marine water than those in freshwater mussels and snail. However, omega-6 fatty acids like linoleic acid (LA), α- linolenic acid (ALA) and arachidonic acid (ARA) were higher in freshwater mussels and snail than in the marine bivalves. The n-3/n-6 ratio were significantly higher in oysters and cockle species than the other groups of bivalves and snail. The index of atherogenicity and index of thrombogenicity of the mollusc species ranged from 0.74 ± 0.1 to 1.74 ± 0.2 and 0.5 ± 0.1 to 2.6 ± 0.2, respectively. The results show that marine water bivalves contained higher amount of potassium, sodium, iron, chlorine especially oyster species contained significantly higher iodine than the freshwater bivalves and snail. However, freshwater mussels and snail showed significantly higher amount of zinc contents than the marine bivalves. The heavy metal contents such as arsenic, chromium and mercury were absent or present in very tiny amounts among the mollusc species. Significantly higher amount of cholesterol was present in marine bivalves and freshwater snail species than the freshwater mussels. Overall, the results indicate that marine bivalves can be good sources of high quality protein and lipid especially EPA and DHA. On the other hand, freshwater mussels and snails also could be good sources of protein, LA and ARA but scarcity of EPA and DHA.

The price of fish oil has reached a historical peak due to a consistent downward production trend, and therefore, the search for sustainable alternative sources has received great attention. This research was conducted to evaluate dietary micro-algae, Schizochytrium sp. (SC) as fish oil (FO) replacer in rainbow trout, Oncorhynchus mykiss. In the first trial, apparent digestibility coefficient (ADC) was 92.4% for dry matter, 91.4% for crude protein, and 94.2% for crude lipid in rainbow trout. In the second trial, six diets were formulated to replace FO at 0% (CON), 20% (T20), 40% (T40), 60% (T60), 80% (T80), and 100% (T100) with SC in the rainbow trout (3.0 ± 0.4 g, mean ± SD) diet. After eight weeks’ feeding trial, weight gain (WG), specific growth rate (SGR), and feed efficiency (FE) of fish fed the T20 diet were significantly higher than those of fish fed other diets (p < 0.05). However, there were no significant differences in these parameters among those of fish fed CON, T40, T60, and T80 diets. Lysozyme activity of fish fed the T20 diet was significantly higher than those of fish fed other experimental diets (p < 0.05). After 10 days of disease challenge testing with pathogenic bacteria (Lactococcus garvieae 1 × 108 CFU/mL), the cumulative survival rate of fish fed the T20 diet was significantly higher than those of fish fed the CON, T80, and T100 diets. Therefore, these results suggest dietary microalgae SC is well-digested and could replace up to 80% of fish oil in the diet of rainbow trout without negative effects on growth and immune responses.

Fishmeal is an important source of protein in the fish diet, but its supply is unstable due to environmental pollution and overfishing of anchovy and herring. In addition, the disease outbreak of fish has been increased, and the abuse of antibiotics and vaccines has been becoming a problem for residual substances. Therefore, studies that address diets with reduced fishmeal content and additives that can increase efficiency and replace antibiotics are needed. Hence, in this study, the probiotics bacteria Bacillus sp. SJ-10 (BSJ-10) and Lactobacillus plantarum (LP) were heat-killed (HK) used as additives in a low-fishmeal diet. The two bacteria with a 45 % fishmeal diet were added at a concentration of 3.34 × 108 CFU/mL, and both of probiotics bacteria were mixed and added to the same concentration in order to produce the diet for HK BSJ-10 (HKBSJ-10), HK LP (HK-LP), and MIX (HKBSJ-10 + HK-LP) groups. We raised four replicates of juvenile olive flounder for 8 weeks. In the HKBSJ-10 group, the plasma insulin, GH, and IGF-I level were higher than the other groups (P

Recent research is increasingly shedding light on the important role that microbial metabolites such as γ-aminobutyric acid (GABA) play in the context of nutrition, cognition, immune function, and the modulation of the gut microbiome. Yet, very few trials were conducted to assess the effects of its supplementation on biomarkers of fish health. Therefore, an eight-week feeding trial was devised to evaluate GABA supplementation in juvenile olive flounder, (Paralichthys olivaceus). A total of 630 fish with an average weight of 4.90 ± 0.10 g (±SD) were randomly assigned to one of seven triplicate groups and fed a non-GABA supplemented diet (CON, with 92 mg/kg GABA content), a positive control with 4 g/kg oxytetracycline (OTC), and five other diets supplemented with 50, 100, 150, 200 and 250 mg/kg GABA (corresponding to a total GABA content of 154, 229, 282, 327 and 352 mg/kg, respectively). Growth, blood chemistry, nonspecific immunity, digestive enzyme activity and disease resistance were assessed. The results showed that 100 and 150 mg/kg GABA supplementation consistently yielded significant improvements (p < 0.05) in growth, intestinal amylase, serum lysozyme, and survival against infection with Streptococcus iniae. Based on polynomial analysis, the optimal supplementation level was determined to be 237 mg/kg. These results support GABA as an important functional feed additive in juvenile olive flounder.

We investigated the three probiotic bacteria and a processed yeast (GroPro-Aqua) as the replacers of antibiotics in juvenile olive flounder. A total of seven diets were used, that is, one basal or control (CON) diet; and six other diets, of which, three diets were prepared by supplementing probiotic bacteria such as Bacillus subtilis WB60 (BSWB60) at 1 × 108 CFU/g diet, Bacillus subtilis SJ10 (BSSJ10) at 1 × 108 CFU/g diet, and Enterococcus faecium SH30 (EFSH30) at 1 × 107 CFU/g diet; one diet with processed yeast (GRO) at 0.35% diet; and two other diets were supplemented with oxytetracycline (OTC) and amoxicillin (AMO) at 4 g/kg of each. Triplicate groups of fish (average 12.1 g) were fed one of the diets for eight weeks. At the end of the feeding trial, the fish that were fed the probiotic bacteria-supplemented diets had a significantly higher final weight, weight gain, and specific growth rate compared to the CON, OTC, and AMO diets. Fish that were fed the GRO diet had significantly higher feed efficiency and protein efficiency ratios than those of the fish that were fed the CON diet. Serum glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, glucose, and total protein were not affected by the diets. Lysozyme activity in fish that were fed the BSSJ10, BSWB60, and EFSH30 diets were significantly higher compared to the CON and OTC diets, whereas myeloperoxidase activity of fish fed the BSWB60 and EFSH30 diets were significantly higher than those of fish fed the CON and AMO diets. Flounder growth hormone gene expressions of fish that were fed BSWB60 and GRO diets were significantly higher compared to the CON, OTC, and AMO diets. The interleukin-1β gene expression of fish that were fed the BSSJ10, BSWB60, EFSH30, OTC, and GRO diets was significantly higher than those of fish fed the CON diet. The interleukin-10 gene expression of fish that were fed the BSSJ10, EFSH30, and GRO diets was significantly higher than those of fish fed the CON and AMO diets. Posterior intestinal histology of fish showed significantly higher villi length in fish that were fed the BSSJ10, BSWB60, EFSH30, and GRO diets compared to the CON diet. After 15 days of challenge test with pathogenic bacteria Edwardsiella tarda, the cumulative survival rate of fish that were fed the BSSJ10, BSWB60, EFSH30, and GRO diets were significantly higher than those of fish that were fed the CON diet. Overall, the results indicate that dietary supplementation of B. subtilis (108 CFU/g diet), E. faecium (107 CFU/g diet), and processed yeast (GroPro-Aqua at 0.35% diet) could replace the antibiotics in terms of improving growth, immunity, gut health, and disease resistance in juvenile olive flounder.

Abstract Three different dietary additives were evaluated based on growth performance, innate immune responses and disease resistance in juvenile Amur catfish Silurus asotus. Four diets were prepared by supplementing 0.4% Song-gang® stone (SG), 0.05% Yucca meal (YM), 0.05% β-glucan (BG) in combination with 0.04% SG and 0.05% BG (SG + BG) to a basal commercial diet (control; CONT). Triplicate groups of fish averaging 4.95 ± 0.05 g were fed one of the test diets between 3 and 5% body weight for 8 weeks. At the end of the feeding trial, weight gain, specific growth rate, feed efficiency and protein efficiency ratio of fish fed SG and SG + BG were significantly higher than those of fish fed the BG and CONT diets (P

A 6-week feeding trial was carried out to evaluate the effects of dietary vitamin E (dl-α-tocopheryl acetate, TA) on growth and mercury (Hg) accumulation in juvenile olive flounder (Paralichthys olivaceus) treated with mercuric chloride (HgCl2). Vitamin E and HgCl2 were added to the semi-purified basal diet. Six semi-purified diets in a 2 × 3 factorial design were formulated to contain 2 levels of Hg (0 or 20 mg HgCl2/kg diet) and 3 levels of vitamin E (0, 100, or 200 mg TA/kg diet). Experimental fish (n = 360, 9.99 ± 0.15 g) were randomly allocated into 30-L tanks at a density of 20 fish per tank with 3 replicates in each treatment and were fed twice a day. At the end of the feeding trial, dietary Hg depressed the growth performances in terms of weight gain (WG), specific growth rate (SGR), feed efficiency (FE) and protein efficiency ratio (PER) in fish, while fish fed the diets supplemented with vitamin E showed significant growth improvement in both presence and absence of HgCl2 in the diets (P

Abstract Different levels of dietary soybean meal (SBM) as a fish meal (FM) replacer, with and without amino acid supplementation, for whiteleg shrimp, Litopenaeus vannamei reared in the biofloc system was examined in eight weeks of feeding trial. Eight experimental diets consisted of a basal diet with 0% FM replacement by SBM provided in clear sea water without biofloc system (S0SW), four diets replacing FM at 0% (S0), 33% (S33), 67% (S67) and 100% (S100) by SBM, and three diets replacing FM at 33% (S33A), 67% (S67A) and 100% (S100A) by SBM supplemented with amino acids (methionine and lysine) in the seawater biofloc system. Results of water quality analyses showed significantly lower total suspended solids and nitrate for S0SW group than all other treatments. Diets S0 and S33A resulted in higher weight gain and specific growth rate among all groups, with no significant differences with S33 group. In addition, whole-body protein and amino acid compositions of shrimp fed S0SW were lower than most biofloc groups. Haemolymph parameters showed significant differences in total protein, cholesterol and triglyceride between groups S0 and S0SW. Also, superoxide dismutase activity showed a decreasing trend with increasing replacement level. In conclusion, based on these results, SBM could replace up to 33% of FM with or without amino acid supplementation in juvenile whiteleg shrimp diets reared in the biofloc system.

We performed an 8-week feeding trial to evaluate dietary soluble extract hydrolysates from fishery by-products, such as shrimp soluble extract (SSE) with or without inosine monophosphate (IMP), tilapia soluble extract (TSE) and squid soluble extract (SQSE), in juvenile Nile tilapia. A diet without feed additives was used as the control diet (CON); and five other experimental diets were formulated with 2% soluble extracts consisting of 100% SSE, 98% SSE + 2% IMP (SSEP2), 96% SSE + 4% IMP (SSEP4), 100% SQSE and 100% TSE. The diets were fed to 4.9 ± 0.07 g (mean ± SD) juvenile Nile tilapia in triplicate groups. The weight gain and specific growth rates of fish fed the SSE, SSEP2 and SSEP4 diets were significantly higher than those of fish fed the CON and SQSE diets. The superoxide dismutase activity levels of fish fed the SSE and SSEP4 diets were significantly higher than those of fish fed the CON, SSEP2, SQSE and TSE diets. Myeloperoxidase activity levels of fish fed the SSE and SSEP4 diets were significantly higher than those of fish fed the CON, SSEP2 and SQSE diets. Lysozyme activity levels of fish fed the SSEP4 and SQSE diets were significantly higher than those of fish fed the SSE and SSEP2 diets. Feed efficiency, protein efficiency ratio, survival rate, whole body proximate composition and hematological parameters were not significantly different among the groups. After ten days of challenge = against Aeromonas hydrophila, the cumulative survival rate of fish fed the SSE diet was significantly higher than those of fish fed the CON, SQSE and TSE diets. In conclusion, dietary shrimp soluble extract could improve the growth performance, non-specific immune responses and disease resistance in juvenile Nile tilapia, and inosine monophosphate did not add further benefits to this ingredient.

Abstract A feeding trial was conducted to evaluate the effects of two different sizes of extruded pellets (EP) (EP1 - 3 mm or EP2 - 5 mm) and a moist pellet (MP) in olive flounder, Paralichthys olivaceus, reared in semi-recirculation system. A total of 450 fish with an average initial weight of 5.0 ± 0.2 g (mean ± SD) were fed one of the three experimental diets in triplicate groups. At the end of a 6-week feeding trial, weight gain, specific growth rate, and feed efficiency of fish fed EP diets were significantly higher than those of fish fed MP (P

Abstract We determined the optimum dietary protein level in juvenile barred knifejaw Oplegnathus fasciatus in cages. Five semi-purified isocaloric diets were formulated with white fish meal and casein-based diets to contain 35, 40, 45, 50, and 60 % crude protein (CP). Fish with an initial body weight of 7.1 ± 0.06 g (mean ± SD) were randomly distributed into 15 net cages (each size: 60 cm × 40 cm × 90 cm, W × L × H) as groups of 20 fish in triplicates. The fish were fed at apparent satiation level twice a day. After 8 weeks of feeding, the weight gain (WG) of fish fed 45, 50, and 60 % CP diets were significantly higher than those of fish fed 35 and 40 % CP diets. However, there were no significant differences in WG among fish fed 45, 50, and 60 % CP diets. Generally, feed efficiency (FE) and specific growth rate (SGR) showed a similar trend as WG. However, the protein efficiency ratio (PER) was inversely related to dietary protein levels. Energy retention efficiency increased with the increase of dietary protein levels by protein sparing from non-protein energy sources. Blood hematocrit content was not affected by dietary protein levels. However, a significantly lower amount of hemoglobin was found in fish fed 35 % CP than in fish fed 40, 45, 50, and 60 % CP diets. Fish fed 60 % CP showed the lowest survival rate than the fish fed 35, 40, 45, and 50 % CP diets. Broken-line analysis of WG showed the optimum dietary protein level was 45.2 % with 18.8 kJ/g diet for juvenile barred knifejaw. This study has potential implication for the successful cage culture of barred knifejaw.

Abstract The present study was conducted to evaluate the effects of dietary protein levels on growth, biometrics, hematology and body composition in juvenile parrot fish Oplegnathus fasciatus. Fish averaging 7.1 ± 0.06 g (mean ± SD) was randomly distributed into 15 net cages (each size: 60 × 40 × 90 cm, W × L × H) as groups of 20 fish. Five isocaloric diets (16.7 kJ/g energy) were formulated to contain crude protein levels (CP) as 35 (CP35), 40 (CP40), 45 (CP45), 50 (CP50) and 60 % (CP60) in the diets. Fish were fed one of the experimental diets at apparent satiation twice a day in triplicate groups. At the end of 8-week feeding trial, weight gain (WG) of fish fed with CP50 and CP60 diets were significantly higher than those of fish fed with CP35, CP40 and CP45 diets. Fish fed with CP45, CP50 and CP60 diets had higher feed efficiency (FE) and specific growth rate (SGR) than those of fish fed with CP35 and CP40 diets. Protein retention efficiency (PRE) decreased with increase of dietary protein levels among fish fed with the experimental diets. Whole-body crude protein and lipid contents increased with the dietary protein level up to CP50 diet. In conclusion, analysis of variance (ANOVA) revealed that the optimum dietary protein level could be 50 % for maximum growth of juvenile parrot fish, while the broken-line analysis of WG suggested that the level could be 48.5 %, in a diet containing 16.7 kJ/g energy.

Abstract This study evaluated the dietary lysine requirement by measuring the plasma free lysine concentrations in rainbow trout, Oncorhynchus mykiss after dorsal aorta cannulation. A basal diet containing 36.6 % crude protein (29.6 % crystalline amino acids mixture, 5 % casein and 2 % gelatin) was formulated to one of the seven L-amino acid based diets containing graded levels of lysine (0.72, 1.12, 1.52, 1.92, 2.32, 2.72 or 3.52 % dry diet). A total of 35 fish averaging 512 ± 6.8 g (mean ± SD) were randomly distributed into seven groups with five fish in each group. After 48 h of feed deprivation, each group of fish was fed one of the experimental diets by intubation at 1 % body weight. Blood samples were taken at 0, 5 and 24 h after intubation. Post-prandial plasma free lysine concentrations (PPlys, 5 h after intubation) of fish fed diets containing ≥ 2.32 % lysine were higher than those of fish fed diets containing ≤ 1.92 % lysine. Post-absorptive free lysine concentrations (PAlys, 24 h after intubation) of fish fed diets containing 2.32 and 3.52 % lysine were higher than those of fish fed diets containing ≤ 1.52 % lysine. The broken-line regression analysis on the basis of PPlys and PAlys indicated that the lysine requirement of rainbow trout could be 2.34 and 2.20 % in diet. Therefore, these results strongly suggested that the dietary lysine requirement based on the broken-line model analyses of PPlys and PAlys could be greater than 2.2 but less than 2.34 % (corresponding to be 6.01 % ≤, but ≤ 6.39 % in dietary protein basis, respectively) in rainbow trout. Also, these results shown that the quantitative estimation of lysine requirement by using PPlys and PAlys could be an acceptable method in fish.

Mercury as one of the most toxic elements can be present in organic or inorganic form in marine fishes, which may cause a potential threat to public health. In this study, we investigated to determine the dietary organic (O-Hg) and inorganic (I-Hg) mercury threshold levels on induced mercury toxicity in juvenile olive flounder, Paralichthys olivaceus as a marine fish model. Twenty-eight fish averaging 3.1 ± 0.05 g (mean ± SD) were arbitrarily assigned to each of 27 tanks. Each tank was arbitrarily restricted to triplicates of nine experimental diets for eight weeks. The experimental diets were manufactured to contain 0 (Control), 10 (I-Hg10, O-Hg10), 20 (I-Hg20, O-Hg20), 40 (I-Hg40, O-Hg40) and 160 (I-Hg160, O-Hg160) mg/kg diet in organic form as methylmercury (MeHg) or in inorganic form as mercuric chloride (HgCl2). At the termination of the experimental trial, weight gains (WGs) of fish fed the control and 10 (I-Hg10, O-Hg10) diets were remarkably higher than those of fish fed the 20 (I-Hg20, O-Hg20), 40 (I-Hg40, O-Hg40) and 160 (I-Hg160, O-Hg160) (p < 0.05). Specific growth rate and feed efficiency of fish fed control and 10 (I-Hg10, O-Hg10) diets were significantly higher than those of fish fed 40 (I-Hg40, O-Hg40) and 160 (I-Hg160, O-Hg160) diets. In comparison to the dietary inorganic mercury, dietary MeHg bioaccumulation rates were significantly higher in the tissue levels according to the dietary inclusion levels. MeHg accumulated mostly in kidney, followed by liver and gill tissues. HgCl2 accumulated in tissues, in decreasing order, liver > kidney > gills. A broken-line regression model for percentage of WG indicated that the threshold toxicity level for an Hg-incorporated diet of juvenile olive flounder could be 13.5 mg Hg/kg in the form of HgCl2 and 8.7 mg Hg/kg in the form of MeHg.

An eight-week feeding trial was conducted to evaluate the effects of different dietary probiotic supplements in juvenile whiteleg shrimp, Litopenaeus vannamei. A basal control diet without probiotics (CON), and five other diets by supplementing Bacillus subtilis at 107 CFU/g diet (BS7), B. subtilis (BS8), Pediococcus pentosaceus (PP8), and Lactococcus lactis (LL8) at 108 CFU/g diet, and oxytetracycline (OTC) at 4 g/kg diet were used. Whiteleg shrimp with initial body weights of 1.41 ± 0.05 g (mean ± SD) were fed with these diets. Growth of shrimp fed BS8 and LL8 diets was significantly higher than those of shrimp fed the CON diet (p < 0.05). Superoxide dismutase activity in shrimp fed PP8 and LL8 diets was significantly higher than that of shrimp fed the CON diet (p < 0.05). Lysozyme activity of shrimp fed probiotics and OTC diets significantly improved compared to those on the CON diet (p < 0.05). The intestinal histology showed healthier guts for shrimp fed the probiotic diets (p < 0.05). Immune-related gene expression in shrimp fed BS8, PP8 and LL8 diets was recorded as significantly higher than that of shrimp fed CON and OTC diets (p < 0.05). Also, results of the challenge test for 7 days and the digestive enzyme activity of shrimp fed BS8, PP8, and LL8 were significantly improved compared to those on the CON diet (p < 0.05). Therefore, these results indicated that L. lactis at 108 CFU/g could be an ideal probiotic for whiteleg shrimp, and also B. subtilis WB60 and P. pentosaceus at 108 CFU/g could improve the growth, immunity, histology, gene expression, digestive enzyme activity, and disease resistance, while replacing antibiotics.

An eight-week feeding trial was conducted to evaluate the effects of a basal control diet (CON), Bacillus subtilis at 107 (BS7) and at 108 CFU/g diet (BS8), Lactococcus lactis at 107 CFU/g (LL7) and at 108 CFU/g diet (LL8), and oxytetracycline (OTC) at 4 g/kg diet on Nile tilapia. Fish with initial body weight of 2.83 ± 0.05 g (mean ± SD) were fed two times a day. Weight gain, specific growth rate, feed efficiency, protein efficiency ratio and lysozyme activity of fish fed BS8, LL8 and LL7 diets were significantly higher than those of fish fed CON diet (p < 0.05). Superoxide dismutase and myeloperoxidase activity of fish fed BS8, LL8, BS7, LL7 and OTC diets were significantly higher than those of fish fed CON diet. Intestinal villi length and muscular layer thickness of fish fed BS8, LL8 and LL7 diets were significantly higher than those of fish fed CON and OTC diets. Also, heat shock protein 70 (HSP70), interleukin (IL-1β), interferon-gamma (IFN-γ) and tumour necrosis factor (TNF-α) gene expression of fish fed BS8 and LL8 diets were significantly higher than those of fish fed CON diet. After 13 days of challenge test, cumulative survival rate of fish fed BS8 and LL8 diets were significantly higher than those of fish fed CON, BS7 and OTC diets. Based on these results, B. subtilis and L. lactis at 108 (CFU/g) could replace antibiotics, and have beneficial effects on growth, immunity, histology, gene expression, and disease resistance in Nile tilapia.

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

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

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

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