Evaluation of Garlic Juice Processing Waste Supplementation in Juvenile Black Rockfish (Sebastes schlegelii) Diets on Growth Performance, Antioxidant and Digestive Enzyme Activity, Growth- and Antioxidant-Related Gene Expression, and Disease Resistance against Streptococcus iniae.

An 8-week feeding trial was conducted to evaluate the effects of various dietary levels of garlic juice processing waste (GJPW) on the growth, feed utilization, digestive and antioxidant enzyme activity, growth- and antioxidant-related gene expression, and resistance to Streptococcus iniae infection of juvenile black rockfish (Sebastes schlegelii). A total of 450 juvenile rockfish were randomly distributed into 30 L rectangular tanks (30 fish per tank). Five experimental diets were prepared in triplicate. The fish were fed experimental diets supplemented with GJPW at concentrations of 0 (GJPW0, control), 2.5 (GJPW2.5), 5 (GJPW5), 7.5 (GJPW7.5), and 10 g kg-1 (GJPW10) diet. All of the GJPW-supplemented treatments (2.5, 5, 7.5, and 10 g kg-1) significantly enhanced weight gain (WG), specific growth rate (SGR), feed efficiency (FE), protein efficiency ratio (PER), and digestive enzyme activity (amylase, trypsin, and lipase). A decreasing trend was seen in plasma aspartate aminotransferase (ALT), alanine aminotransferase (AST), and glucose (GLU) content with increasing dietary levels of GJPW. In contrast, plasma lysozyme and antioxidant enzyme activities were significantly increased with increasing dietary GJPW levels. Furthermore, GJPW administration significantly upregulated the expression of insulin-like growth factor-1 (IGF-1), superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) in the liver of rockfish. A challenge test with S. iniae showed significantly higher resistance in the GJPW-supplemented treatments than in the control. In short, dietary supplementation GJPW enhanced growth performance and antioxidant response in juvenile black rockfish, with suitable effects in fish fed with 2.5 g kg-1 GJPW for 8 weeks.

Dietary Supplementation with Ginger (Zingiber officinale) Residue from Juice Extraction Improves Juvenile Black Rockfish (Sebastes schlegelii) Growth Performance, Antioxidant Enzyme Activity and Resistance to Streptococcus iniae Infection.

Plant-derived feed additives provide cost effective and environmentally friendly alternatives to antibiotics for improving fish performance in aquaculture. An 8-week feeding trial was conducted to evaluate the effects of dietary ginger residue from juice extraction (GRJE) on juvenile black rockfish (Sebastes schlegelii) growth performance, antioxidant enzyme activities, and resistance to Streptococcus iniae infection. Juvenile rockfish (n = 450; initial weight = 2.2 ± 0.01 g) were randomly distributed into 30 L rectangular tanks (30 fish per tank). Five experimental diets with GRJE concentrations of 0% (control), 0.25%, 0.5%, 0.75%, and 1% were prepared in triplicate. Three groups of fish were randomly assigned to each diet and fed to apparent satiation twice daily. After the feeding trial, fish were challenged with S. iniae, and cumulative survival was observed for six days. Growth parameters, feed efficiency, and the protein efficiency ratio showed a quadratic correlation with the GRJE concentration in the fish diet. Proximate composition and plasma chemistry were not significantly affected. Plasma lysozyme, superoxide dismutase, glutathione, and catalase activities linearly increased with increasing GRJE supplementation levels. Moreover, survival in the S. iniae challenge test was significantly higher in fish fed diets supplemented with 0.75-1% GRJE. Our findings demonstrated that 0.75% GRJE dietary supplementation enhanced the growth performance, antioxidant activity, and disease resistance of juvenile black rockfish with no adverse effects.

Internal dynamics of inorganic and methylmercury in a marine fish: Insights from mercury stable isotopes.

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 (∼0.63‰) 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.

A comparative study of effects of dietary mercuric chloride and methylmercury chloride on growth performance, tissue accumulation, stress and immune responses, and plasma measurements in Korean rockfish, Sebastes schlegeli.

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.

Dietary effect of low fish meal aquafeed on gut microbiota in olive flounder (Paralichthys olivaceus) at different growth stages.

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.

Effects of Dietary Protein and Lipid Levels on Growth and Body Composition of Juvenile Far Eastern Catfish Silurus asotus.

A 3×2 factorial experiment was conducted to determine the effects of dietary protein and lipid levels on the growth and body composition of juvenile far eastern catfish. Six diets were formulated to contain three levels of protein (20%, 30% and 40%) and two levels of lipid (9% and 17%). Triplicate groups of fish (initial body weight of 7.6 g) were hand-fed to apparent satiation for 66 days. Final mean weight was improved with increasing dietary protein and lipid levels, and the highest final mean weight was observed in fish fed the 40/17 (% protein/% lipid) diet. No significant difference was observed in final mean weight for fish fed between 30/17 diet and 40/9 diet. Feed efficiency of fish fed the diets containing over 30% protein levels with 9% and 17% lipid levels were significantly higher than those of fish fed the 20% protein levels. Feed efficiency of fish fed the 30/17 diet was not significantly different from that of fish fed the 40/9 diet or 40/17 diet. Feed efficiency and protein efficiency ratio of fish fed the 20% protein diets with 17% lipid level were significantly higher than those of fish fed 9% lipid diet. Daily feed intake of fish tended to decrease with increasing dietary protein and lipid levels. Moisture content of whole body in fish fed the 9% lipid diets was significantly higher than that of fish fed the 17% lipid diets at the same protein level, but the opposite trends were found for crude lipid content. Significant effects of dietary lipid were observed for most fatty acids, according to their relative values in the diets. The results of this study suggest that the protein requirement for maximum growth of juvenile far eastern catfish may be higher than 40%, and an increase of dietary lipid level from 9% to 17% can improve growth and feed utilization.

Effects of Dietary Lipid Source and Level on Growth Performance, Blood Parameters and Flesh Quality of Sub-adult Olive Flounder (Paralichthys olivaceus).

This study was conducted to investigate the effects of dietary lipid source and level on growth performance, blood parameters, fatty acid composition and flesh quality of sub-adult olive flounder Paralichthys olivaceus. Eight experimental diets were formulated to contain 5% squid liver oil (SLO), 5% linseed oil (LO), 5% soybean oil (SO), a mixture of 1% squid liver oil, 2% linseed oil and 2% soybean oil (MIX), no lipid supplementation with high protein level (LL-HP), 10% squid liver oil (HL-SLO), a mixture of 1% squid liver oil, 4.5% linseed oil and 4.5% soybean oil (HL-VO), and 1% squid liver oil with high starch level (LL-HC), respectively. Two replicate groups of fish (average initial weight of 296 g) were fed the diets for 17 wks. After 5 wks, 11 wks and the end of the feeding trial, five fish from each tank were randomly sampled for analysis of body composition. At the end of the feeding trial, final mean weight of fish fed the LL-HP diet was significantly (p<0.05) higher than that of fish fed the HL-VO diet, but did not differ significantly from those of fish fed the SLO, LO, SO, MIX, HL-SLO and LL-HC diets. Fish fed the LL-HP diet showed significantly higher feed efficiency than fish fed the LO, HL-SLO and HL-VO diets. Feed efficiency of fish fed the LO, SO and MIX diets were similar to those of fish fed the SLO and HL-SLO diets. Fish fed the HL-SLO diet showed significantly higher total cholesterol content in plasma compared with other diets. Fatty acid composition of tissues was reflected by dietary fatty acid composition. The highest linoleic (LA) and linolenic acid (LNA) contents in the dorsal muscle were observed in fish fed the SO and LO diets, respectively, regardless of feeding period. The highest eicosapentaenoic acid (EPA) content in the dorsal muscle was observed in fish fed the LL-HP and LL-HC diets after 11 and 17 weeks of feeding, respectively. Fish fed the SLO and HL-SLO diets showed higher docosahexaenoic acid (DHA) content than that of other treatments after 11 and 17 weeks of feeding, respectively. Dietary inclusion of vegetable oils reduced n-3 HUFA contents in the dorsal muscle and liver of fish. The n-3 HUFA contents in tissues of fish fed the SLO and HL-SLO diets were higher than those of fish fed other diets, except for the LL-HP and LL-HC diets. Hardness, gel strength, chewiness and cohesiveness values of dorsal muscle in fish were significantly affected by dietary lipid source. The results of this study indicate that fish oil in fish meal based diets for sub-adult olive flounder could be replaced by soybean oil and linseed oil without negative effects on growth and feed utilization.