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The experiment was conducted to evaluate the supplementary effects of gamma aminobutyric acid (GABA) and sodium butyrate (SB) when a graded level of fish meal (FM) was replaced with soy protein concentrate (SPC) in diets for juvenile red seabream (Pagrus major). A control diet was designed to contain 60% FM (F60). Two other diets were formulated by reducing FM levels to 40% and 20% with SPC (F40 and F20). Six more diets were formulated by adding 0.02% GABA or 0.2% SB to each F60, F40 and F20 diets (F60G, F60S, F40G, F40S, F20G and F20S). Each diet was randomly assigned to a triplicate group of fish (5.52 g/fish) and provided for eight weeks. Final body weight, weight gain and specific growth rate of fish fed F60G, F60S, F40G and F40S diets were comparable and significantly higher (p < 0.05) than other groups. The growth of fish fed SB-containing diets was significantly increased (p < 0.05) compared to fish fed the respective control diets. The feed efficiency and protein efficiency ratios were significantly higher (p < 0.05) in the fish fed all diets containing 60% and 40% FM compared to F20 and F20G groups. The F40S diet resulted in the highest feed utilization values. The F20S group exhibited significantly higher (p < 0.05) feed utilization than the F20 and F20G groups. Serum lysozyme activity was significantly higher (p < 0.05) in fish fed the GABA- and SB-containing diets compared to the F20 group. The F60S group exhibited the highest lysozyme activity which was significantly higher (p < 0.05) than in the F20 and F40 groups. Therefore, the growth performance, feed utilization and innate immunity of red seabream can be enhanced by dietary supplementation with GABA or SB in low-FM diets containing SPC. The FM level in the juvenile red seabream diet can be reduced to 40% with SPC and GABA or SB while maintaining performance better than a diet containing 60% FM.
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Lipids in fish diets provide energy and play important roles in immunity and metabolism. Atlantic salmon, a species that migrates from freshwater to seawater, requires high energy, especially during smoltification. Juvenile teleosts have low lipid requirements, and a high dietary lipid content is known to have negative effects on their growth and digestion. Therefore, this study evaluated the effect of two commercial rainbow trout feeds (low-lipid, 13.41% and 14.6%) on the growth and immune responses of early parr-stage Atlantic salmon compared to commercial salmon feed (high-lipid, 29.52%). Atlantic salmon parr (weight: 14.56 ± 2.1 g; length: 11.23 ± 0.44 cm) were randomly divided into three groups and fed either one of two commercial rainbow trout feeds (RTF1 and RTF2) or the commercial salmon feed (ASF) for 12 weeks. At the end of the feeding trial, growth, haematology, histology and gene expression analyses were performed. There were no significant differences in weight gain rates or feed efficiency between the groups (p > 0.05). Superoxidate dismutase, glutathione peroxidase, lysozyme and immunoglobulin M activities were not different among the experimental groups (p > 0.05). A histological examination of the liver and intestinal tissues showed no pathological symptoms of inflammatory response or lipid accumulation in any of the groups. In an intestinal transcriptome analysis using RNA-seq, the expression levels of several genes linked to lipids, immune-related proteins, cytokines and chemokines did not differ significantly between the groups (p > 0.05). Commercial rainbow trout feed with low lipid content has no clear negative impact on the development of Atlantic salmon during the early parr stage (14.5 to 39.6 g). This study provides basic information for the development of economical feed for early parr-stage Atlantic salmon.
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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.
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This study investigated the effects of supplementation of low-temperature probiotics isolated from the intestines of olive flounder on the growth performance, digestibility, and regulation of intestinal microbiota and the expression of genes related to growth, immunity, and apoptosis in olive flounder. Bacteria showing high growth at approximately 15-20 °C, which is the temperature of olive flounder culture, were isolated and confirmed to be Pseudomonas species through 16S rRNA gene sequence analysis. Whole-genome sequencing revealed that the strain has a 6,195,122 bp single circular chromosome and a guanine-cytosine content of 59.9%. In the feeding trial, supplementation with 1 × 108 CFU/g of the isolate strain positively modulated growth performances, digestive enzyme activity, and gut microbiota composition of olive flounder. RT-qPCR for the comparison of growth, immunity, and apoptosis-related gene expression levels showed no significant differences between the groups. Therefore, the isolated host-associated low-temperature probiotics improved the growth performance of olive flounder by causing positive changes in digestive activity and intestinal microbial composition without affecting host gene expression.
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Enfermedades de los Peces , Lenguado , Probióticos , Animales , Acuicultura , Enfermedades de los Peces/microbiología , Probióticos/farmacología , ARN Ribosómico 16S/genética , TemperaturaRESUMEN
In this study, a 12-week feeding experiment was conducted to characterize the effects of exogenous α-amylase on the growth, feed utilization, digestibility, plasma α-amylase activity, feed degradation rate, and fecal particle size of olive flounder (Paralichthys olivaceus). Diet was supplemented with 0 (AA0; control), 100 (AA100), 200 (AA200), or 400 (AA400) mg/kg of α-amylase, respectively. Fish (273.1 ± 2.3 g) were stocked into 12 tanks (25 fish/1,000-L tank) and 3 tanks were randomly selected for each diet group. As a result, α-amylase was found to have no significant effects (p ≥ 0.05) on the growth, feed utilization parameters, and whole-body proximate compositions. α-Amylase-treated fish exhibited only a significant increase in the apparent digestibility coefficient of carbohydrates compared to the controls. In addition, in vitro analyses revealed that α-amylase dose-dependently increased (p < 0.05) the feed degradation rate, while photographs of the intestinal content after 2, 4, and 8 h of feeding demonstrated an improved degradation rate in the α-amylase-treated groups. Plasma α-amylase content was higher in the AA200 and AA400 groups, whereas the control group produced significantly larger-sized fecal particles (90% size class) than these two groups. In the intestine, no changes were observed in the expression levels of the immune-related TNF-α, IL-1ß, IL-2, immunoglobulin-M, HSP-70, lysozyme, and amylase alpha-2A. However, growth-related genes IGF-1, IGF-2, TGF-ß3, and growth hormone genes were upregulated in muscle tissues. Collectively, exogenous α-amylase has positive roles in the modulation of the digestibility coefficient, blood α-amylase concentration, growth-related gene expression, and diet degradation for improved digestion in olive flounder.
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Lenguado , Animales , Inmunidad Innata , alfa-Amilasas , Dieta/veterinaria , Nutrientes , Expresión Génica , Alimentación Animal/análisisRESUMEN
The experiment was conducted to evaluate alternative protein ingredients in a low-fish meal (FM) diet for red seabream (Pagrus major). Twelve experimental diets were formulated. Control diet (CON) was designed to contain 60% FM. Other experimental diets were formulated by replacing 50% of FM from the CON with soy protein concentrate (SPC), corn gluten (CG), meat meal (MM), and/or chicken byproduct meal (CBM). Four diets were designed including one of SPC, CG, MM, or CBM as FM replacer and designated as SPC, CG, MM, and CBM. Six other diets were formulated by adding two ingredients as SPC and CG, SPC and MM, SPC and CBM, CG and MM, CG and CBM, or MM and CBM, and designated as SCG, SMM, SCM, CMM, CCM, and MCM, respectively. The 12th diet (MIX) was formulated by including SPC, CGM, MM, and CBM. Triplicate fish groups (50.2 ± 0.1 g) were hand-fed for 12 weeks. Weight gain (WG) of fish was significantly improved by MM and MCM diets compared to CG, SCG, CMM, and CCM diets. WG of CON, SPC, CM, SMM, SCM, and MIX groups were comparable with MM and MCM groups. The lowest WG was observed in CG and CMM groups. Feed efficiency (FE) was significantly higher in MM group compared to SPC, CG, SGC, and CMC groups. FE of MCM group was significantly higher than CG and SCG groups. Fillet linolenic acid (C18:2n-6) level in CG group was significantly higher than CON, MM, CM, SCM, CCM, and MCM groups. Serum lysozyme activity was significantly higher in MCM and MIX groups. Therefore, a high level of dietary CG reduces the growth performance and feed utilization of red seabream. A mixture of MM and CBM seems to be more efficient in replacing FM from red seabream diet.
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This study investigated the morphological, histological, and histochemical characteristics of the digestive tract of the marbled flounder (Pseudopleuronectes yokohamae). The relative length of the gut of the marbled flounder digestive tract was 1.54 ± 0.10 (n = 20), and it had a simple stomach and 6-9 pyloric caeca. The mucosal folds of the marbled flounder digestive tract exhibited a general branched morphology. The thickness and mucosal fold length of the intestinal muscularis externa showed similar aspects in all areas. The thickness of the intestinal muscularis externa was the thickest in the posterior intestine portion, and the length of mucosal folds was the longest in the anterior intestine portion. It was indicated that food digested by gastric acid in the stomach moves to the anterior portion (including pyloric caeca) and mid portion of the intestine, ensuring effective stimulation of cholecystokinin (CCK)-producing cells. In addition, the distribution pattern of CCK-producing cells in the intestine was very similar to that of mucus-secreting goblet cells. The CCK-producing cells and goblet cells in the marbled flounder were well-adapted to promote optimal control of the digestive process. Based on the morphological and histochemical studies, it was concluded that the marbled flounder displays a digestive tract comparable to that of fish species with carnivorous habits.
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Butylated hydroxytoluene (BHT) is a commonly used antioxidant added to animal/fish feed to limit lipid autoxidation and peroxidation. Although there have been reviews and reports of BHT toxicity in animals, limited information is available with respect to the toxic effects and accumulation of BHT due to oral exposure in aquaculture species. Therefore, 120 days of feeding trial was conducted to evaluate the effects of dietary BHT on the marine fish olive flounder Paralichthys olivaceus. Graded levels of BHT were added to the basal diet in increments of 0, 10, 20, 40, 80, and 160 mg BHT/kg, corresponding to 0 (BHT0), 11 (BHT11), 19 (BHT19), 35 (BHT35), 85 (BHT85), and 121 (BHT121) mg BHT/kg diets, respectively. Fish with an average weight of 77.5 ± 0.3 g (mean ± SD) were fed one of the six experimental diets in triplicate groups. Growth performance, feed utilization, and survival rate were not significantly affected by the dietary BHT levels among all experimental groups, whereas BHT concentration in the muscle tissue was found to increase in a dose-dependent manner up to 60 days of the experimental period. Thereafter, BHT accumulation in muscle tissue showed a declining trend among all treatment groups. Furthermore, the whole-body proximate composition, nonspecific immune responses, and hematological parameters (except triglycerides) were not significantly influenced by the dietary levels of BHT. Blood triglyceride content was significantly higher in fish fed the BHT-free diet compared to all other treatment groups. Thus, this study demonstrates that dietary BHT (up to 121 mg/kg) is a safe and effective antioxidant without exhibiting any adverse effects on the growth performance, body composition, and immune responses in the marine fish olive flounder, P. olivaceus.
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Host-associated probiotics (HAPs) are bacteria originally isolated from rearing water or the host's gastrointestinal tract in order to enhance the host's growth and health. This study investigated the HAP potential of Bacillus sp. PM8313, isolated from wild red sea bream (Pagrus major), through characterization and feeding trials. Results based on in vitro tests showed that PM8313 is safe, confirming its hemolytic, cytotoxic, and antibiotic resistance. In addition, PM8313 showed advantages as a probiotic with high viability in the gastrointestinal model and a high cell adhesion rate. Whole-genome sequencing demonstrated that PM8313 has a 4,615,871 bp single circular chromosome and a guanine-cytosine content of 45.25%. It also showed the absence of genes encoding virulence factors, such as cytotoxin, enterotoxin, hemolysin, sphingomyelinase, and phospholipase. In the feeding trial, a supplemental diet of 1 × 108 CFU/g PM8313 positively altered the weight gain, digestive enzyme activity, and intestinal microbiota composition of red sea bream. Analysis of nonspecific immune parameters and immune-related gene expression, and a challenge test showed that PM8313 supplementation increases immunity and pathogenic bacteria resistance. Our findings suggest that PM8313 should be considered for application as a novel HAP to red sea bream aquaculture.
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A 56-day feeding trial was conducted to determine the effect of dietary supplementation with Bacillus sp. isolated from the intestines of red sea bream on the growth performance, immunity, and gut microbiome composition of red sea bream. Three diets (a control diet and two treatments) were formulated without Bacillus sp. PM8313 or ß-glucan (control, CD), 1 × 108 CFU g-1 PM8313 (BSD), and 1 × 108 CFU g-1 PM8313 + 0.1% ß-glucan (BGSD). At the end of the experiment, the weight, specific growth rate, feed conversion ratio, and protein efficiency ratio of the fish in the BSD and BGSD diet groups were significantly improved than those of the control group (P < 0.05). Additionally, amylase and trypsin activities were significantly higher (P < 0.05) in both groups compared to the control. Superoxide dismutase and lysozyme activity, which are serum non-specific immune responses, only increased in the BGSD group. The two treatment groups exhibited a marked difference in the intestinal microbiota composition compared to the control group. Furthermore, the treatment groups exhibited an upregulation of IL-6 and NF-κb, coupled with high survival rates when challenged with Edwardsiella tarda. Therefore, dietary supplementation with PM8313 improved the growth performance, digestive enzyme activity, non-specific immunity, and pathogen resistance of red sea bream, in addition to affecting the composition of its intestinal microflora.
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Bacillus , Microbioma Gastrointestinal , Perciformes , Dorada , beta-Glucanos , Animales , Alimentación Animal/análisis , beta-Glucanos/farmacología , Suplementos Dietéticos/análisis , Resistencia a la EnfermedadRESUMEN
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.
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The purpose of this study was to characterize the bacteria isolated from rockfish intestines and to investigate the effects of feed supplementation in rockfish aquaculture. Bacillus sp. KRF-7 isolated from the intestine of rockfish (Sebastes schlegelii) was demonstrated to be safe based on in vitro tests confirming the absence of hemolysis, cytotoxicity, and genes with toxigenic potential. In a feeding trial, providing a supplemental diet of 1 × 108 CFU g-1Bacillus sp. KRF-7 was observed to positively alter the weight gain, specific growth rate, feed conversion ratio, and protein efficiency ratio of juvenile rockfish. KRF-7 supplementation showed positive regulation of nonspecific immune parameters, such as superoxide dismutase, lysozyme activity, and myeloperoxidase activity. This analysis also revealed a change in the composition of the intestinal microbiota at the phylum level from Proteobacteria to Firmicutes. In both the kidney and spleen, the expression levels of IL-10, NF-κB, and B cell activating factors in the KRF-7-supplemented group were significantly increased compared to those in the control group. Therefore, this study verified the safety of KRF-7 isolated from the intestine of rockfish and suggests that dietary supplementation with KRF-7 enhances the growth performance of rockfish and has beneficial effects on the regulation of the intestinal microbiota and immune response.
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Bacillus , Lubina , Probióticos , Alimentación Animal/análisis , Animales , Acuicultura , Dieta/veterinaria , Suplementos Dietéticos , Intestinos , Mananos , OligosacáridosRESUMEN
An 8-week feeding trial was conducted to determine the optimal dietary protein level for juvenile marbled flounder. Five semi-purified test diets were formulated to contain different protein levels (CP) including 42.7%, 47.4%, 53.3%, 58.8%, and 64.5% (dry matter), named as CP42.7, CP47.4, CP53.3, CP58.8, and CP64.5, respectively. Five hundred and twenty-five juveniles (6.0 ± 0.1 g) were randomly distributed into 15 tanks (300 L tanks), resulting in 35 fish per tank (n = 3 tanks). Fish were fed the test diets 5 times per day until satiation. The CP58.8 resulted in the highest gain in weight and the best efficiency in feed utilization among the tested protein levels (P < 0.05). Fish fed the CP58.8 diet showed significantly higher whole-body protein and lipid contents than the fish that were fed the other diets (P < 0.05). Fish fed the CP53.3, CP58.8, and CP64.5 diets showed a significantly higher dorsal-muscle lipid content than the fish that were fed the CP42.7 and CP47.4 diets (P < 0.05). The one-slope straight broken-line regression analysis on the results of the thermal growth coefficient and feed conversion ratio indicated that the estimated optimum dietary protein level was 58.8%. Taken together, it is suggested that the dietary protein level of 58.8% is optimal for better growth and high efficiency in feed utilization for the juvenile marbled flounder.
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Experiments were conducted to identify different ratios of Bacillus sp. SJ-10 and Lactobacillus plantarum KCCM 11322 mixtures at a concentration of 1 × 108 CFU/g diet; the effects on growth and cellular and humoral immune responses and the characteristics of disease protection in olive flounder (Paralichthys olivaceus). Flounder were divided into six groups and fed control diet D-1 (without Bacillus sp. SJ-10 and L. plantarum KCCM 11322), positive control diets D-2 (Bacillus sp. SJ-10 at 1 × 108 CFU/g feed) and D-3 (L. plantarum KCCM 11322 at 1 × 108 CFU/g feed); or treatment diets D-4 (3:1 Bacillus sp. SJ-10 and L. plantarum KCCM 11322 at 0.75 + 0.25 × 108 CFU/g feed), D-5 (1:1 Bacillus sp. SJ-10 and L. plantarum KCCM 11322 at 0.50 + 0.50 × 108 CFU/g feed), or D-6 (1:3 Bacillus sp. SJ-10 and L. plantarum KCCM 11322 at 0.25 + 0.75 × 108 CFU/g feed) for 8 weeks. Group D-4 demonstrated better growth and feed utilization (P < 0.05) compared with the controls and positive controls. Similar modulation was also observed in respiratory burst for all treatments and in the expression levels of TNF-α, IL-1ß, IL-6, and IL-10 in different organs in D-4. D-4 and D-5 increased respiratory burst, superoxide dismutase, lysozyme, and myeloperoxidase activities compared with the controls, and only D-4 increased microvilli length. When challenged with 1 × 108 CFU/mL Streptococcus iniae, the fish in the D-4 and D-5 groups survived up to 14 days, whereas the fish in the other groups reached 100% mortality at 11.50 days. Collectively, a ratio-specific Bacillus sp. SJ-10 and L. plantarum KCCM 11322 mixture (3:1) was associated with elevated growth, innate immunity, and streptococcosis resistance (3:1 and 1:1) compared with the control and single probiotic diets.
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Bacillus , Suplementos Dietéticos , Lenguado , Inmunidad Humoral , Lactobacillus plantarum , Alimentación Animal , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Lenguado/crecimiento & desarrollo , Lenguado/inmunologíaRESUMEN
In recent years, considerable and growing attention has been given to the application of host-associated microorganisms as a more suitable source of probiotics in aquaculture sector. Herein, we isolated and screened the olive flounder gut microbiota for beneficial bacterial strains that might serve as potential probiotics in a low fishmeal extruded aquafeed. Among the ten identified isolates, Bacillus amyloliquefaciens SK4079 and B. subtilis SK4082 were screened out based on their heat-resistant ability as well as enzymatic and non-hemolytic activities. Although both strains were well able to utilize carboxymethyl cellulose (CMC), xylan, and soybean meal (SBM) as a single carbon source in the minimal nutrient M9 medium, B. subtilis exhibited significantly higher cellulase, xylanase, and protease activities than B. amyloliquefaciens. The two selected strains were well able to degrade the undesirable anti-nutritional component of the SBM, which would limit its utilization as protein source in aquafeed industry. Significantly higher biofilm formation capacity and notably stronger adhesive interactions with the flounder's skin mucus were detected in B. subtilis than B. amyloliquefaciens. Immobilization of the spores from the selected strains, in a SBM complex carrier, remarkably enhances their thermal resistance at 120 °C for 5 min and different drying conditions. It was also interesting to learn that the B. subtilis spores could survive and remain viable after being sprayed onto extruded low-fish meal feed pellets for as long as 6 months. Overall, the findings of the present study could help the food/feed industries achieve their goal of developing cost-effective yet efficient products.
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Lenguado , Microbioma Gastrointestinal , Probióticos , Animales , Acuicultura , Lenguado/microbiologíaRESUMEN
Mercury isotope ratios in fish tissues have been used to infer sources and biogeochemical processes of mercury in aquatic ecosystems. More experimental studies are however needed to understand the internal dynamics of mercury isotopes and to further assess the feasibility of using fish mercury isotope ratios as a monitoring tool. We exposed Olive flounder (Paralichthys olivaceus) to food pellets spiked with varying concentrations (400, 1600 ng/g) of methylmercury (MeHg) and inorganic mercury (IHg) for 10 weeks. Total mercury (THg), MeHg concentrations, and mercury isotope ratios (δ202Hg, Δ199Hg, Δ200Hg) were measured in the muscle, liver, kidney, and intestine of fish. Fish fed mercury unamended food pellets and MeHg amended food pellets showed absence of internal δ202Hg and Δ199Hg fractionation in all tissue type. For fish fed IHg food pellets, the δ202Hg and Δ199Hg values of intestine equilibrated to those of the IHg food pellets. Kidney, muscle, and liver exhibited varying degrees of isotopic mixing toward the IHg food pellets, consistent with the degree of IHg bioaccumulation. Liver showed additional positive δ202Hg shifts (â¼0.63) from the binary mixing line between the unamended food pellets and IHg food pellets, which we attribute to redistribution or biliary excretion of liver IHg with a lower δ202Hg to other tissues. Significant δ202Hg fractionation in the liver and incomplete isotopic equilibration in the muscle indicate that these tissues may not be suitable for source monitoring at sites heavily polluted by IHg. Instead, fish intestine appears to be a more suitable proxy for identifying IHg sources. The results from our study are essential for determining the appropriate fish tissues for monitoring environmental sources of IHg and MeHg.
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Mercurio , Compuestos de Metilmercurio , Contaminantes Químicos del Agua , Animales , Ecosistema , Monitoreo del Ambiente , Peces , Isótopos de MercurioRESUMEN
Mercury (Hg) is known as a highly toxic heavy metal, and its toxicity varies depending on its form due to different toxicokinetics between inorganic and organic Hg. Limited information on comparison of Hg toxicity concerning its chemical form by oral exposure is currently available in cultured fishes. Therefore, we conducted a comparative study to have a better understanding of distinct toxic effects between mercuric chloride (HgCl2) and methylmercury chloride (CH3HgCl) in Korean rockfish. The 12-weeks dietary exposure of HgCl2 and CH3HgCl with its graded levels (0.4-6.4 ppm) (2 × 5 factorial design) in the young-of-the-year rockfish (initial weight: 82 ± 0.3 g) resulted in neither interactive nor main effects on whole-organism responses, including growth, feed utilization, and survival. However, the distinct pattern of Hg accumulation between the two forms in dorsal muscle, brain, liver and kidney tissues was observed, showing that the rockfish fed the CH3HgCl-contained diets exhibited the dose-dependent accumulation throughout the sampling points (1, 2, 4, 8, and 12 weeks post feeding), whereas those fed the HgCl2-contained diets did not show such response. The CH3HgCl exposure induced higher oxidative stress and immunotoxicity, reflected by the elevated plasma superoxide dismutase and lysozyme activities, respectively. In addition, the CH3HgCl-induced alteration in plasma measurements, including the plasma aspartate transaminase activity and total protein level was found. Taken together, the dietary exposure of methylmercury chloride had more pronounced toxic effects than mercuric chloride in the young-of-year rockfish, needed to be taken into consideration for regulation of maximum allowed levels for Hg by its chemical form.
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Cloruro de Mercurio/toxicidad , Compuestos de Metilmercurio/toxicidad , Perciformes/fisiología , Contaminantes Químicos del Agua/toxicidad , Animales , Encéfalo/metabolismo , Dieta , Mercurio/metabolismo , Estrés Oxidativo , Perciformes/metabolismo , República de Corea , Superóxido Dismutasa/metabolismoRESUMEN
This study was conducted to investigate the long-term effect of a low fish meal (FM) diet comprising plant-based protein sources (PPS) on changes of gut microbial diversity in olive flounder (Paralichthys olivaceus) over the course of life. Two experimental diets were prepared to contain 74% FM (control) or 52% FM with 22% PPS (30% FM replacement, FM30). Fish were fed one of the two experimental diets for 8 months, and we collected the midgut contents to analyze the gut bacterial community by Illumina MiSeq based on the metagenomic sequences in the V3-V4 regions of 16S rRNA. We found that there were nine dominant phyla, which in turn presented Proteobacteria, Firmicutes, and Actinobacteria as the three major phyla in the gut microbiota of the flounder. At genus level, the dominant genera were Delftia, Prevotella, and Chthoniobacter at the juvenile stage (below 100 g/fish); Chthoniobacter, Bacillus, and Bradyrhizobium at the grower stage (400 g/fish); Chthoniobacter, Bacillus, and Delftia at the subadult stage (800 g/fish); and Lactobacillus and Prevotella at the adult stage (over 1,000 g/fish). The microbial diversity in olive flounders arched from the juvenile and subadult stage and reached a plateau thereafter. The fish fed the FM30 diet significantly had an increased abundance of Lactobacillus and Photobacterium and had less abundance of Prevotella and Paraprevotella than the control. However, the effect of dietary PPS was not significant on total microbial richness, indicating no negative effect as feed sources on the intestinal microbiota in olive flounder. These results indicate that the life stage of olive flounder is more important in modulating intestinal microbiota than is the diet. It could also be concluded that dietary PPS might be used as a potential fish meal alternative without any compromising effects on microbial diversity of olive flounder for long-term feeding.
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Alimentación Animal , Lenguado/crecimiento & desarrollo , Lenguado/microbiología , Microbioma Gastrointestinal , Animales , Biodiversidad , Peces , Metagenómica , ARN Ribosómico 16SRESUMEN
A 12-week feeding trial was conducted to evaluate the effects of multi-strain probiotics (MSP) in a low fish meal (FM) diet on overall performance, gut microbiota, selected non-specific immune responses and antioxidant enzyme activities of olive flounder (Paralichthys olivaceus) juveniles. A total of 225 healthy olive flounders (initial mean body weight, 13.5⯱â¯0.01â¯g) were randomly separated into 3 groups of 75 fish, each group having three replicates of 25 fish; first group was fed with a FM-based control diet (Con), 2nd group was fed with a low-FM diet containing a blend of plant and animal protein meals replacing 30% of the FM protein (FM30), and 3rd group was fed with the FM30 diet supplemented with 108-109â¯CFUâ¯kg-1 of the MSP (Pro). With the exception of lipid retention, which was significantly lower in fish fed the FM30 diet compared to the other two treatments, no other statistically significant differences were recorded with respect to any of the other growth and nutrient utilization parameters. Myeloperoxidase and lysozyme activities of fish fed the Pro diet were much higher and significantly different than those of fish fed the FM30 diet. Glutathione peroxidase activity was significantly higher in Pro- than in Con-fed fish, which, in turn, was significantly higher than FM30-fed fish. Expression of immune-related genes including IL-1ß, IL-6, and TNF-α was markedly upregulated in livers of the fish fed Pro diet compared to those fed the Con and FM30 diets. Furthermore, supplementation of MSP in FM30 diet enriched the Lactobacillus abundance in the fish gut as well as predictive gene functions in relation to lipid and carbohydrate metabolisms. These data suggested that the MSP could reduce the potential adverse effects of the low-FM diet and might be used as a healthy immunostimulant for olive flounder.
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Peces Planos/inmunología , Microbioma Gastrointestinal/fisiología , Inmunidad Innata/efectos de los fármacos , Nutrientes/metabolismo , Probióticos/farmacología , Alimentación Animal/análisis , Animales , Dieta/veterinaria , Peces Planos/crecimiento & desarrollo , Peces Planos/metabolismo , Peces Planos/microbiología , Microbioma Gastrointestinal/efectos de los fármacos , Distribución AleatoriaRESUMEN
Investigations were carried out to evaluate and quantify the effects of dietary supplementation with heat-killed (HK) Bacillus sp. SJ-10 (BSJ-10) probiotic (1â¯×â¯108â¯CFUâ¯g-1) on the growth and immunity of olive flounder (Paralichthys olivaceus). Flounder (averagely 9.64â¯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 (Pâ¯<â¯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 (Pâ¯<â¯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 (1â¯×â¯108â¯CFUâ¯mL-1), the HKBSJ-10-fed group exhibited significantly higher protection (Pâ¯<â¯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.