• Fisheries and Aquatic Sciences
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• v.6 no.4
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• pp.209-212
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• 2003

Lucigenin (Lg)- and luminol (Lm)-dependent chemiluminescence (CL) was used to compare the respiratory burst of rockfish (Sebastes schlegeli) phagocytes after stimulation with phorbol myristate acetate (PMA). To establish which reactive oxygen species (ROS) contributes to the observed CL, the modulators of ROS metabolism, such as superoxide dismutase (SOD), catalase, and sodium azide $(NaN_3)$ were used. Although LgCL responses were inhibited significantly by the addition of either SOD or catalase, in comparison to the control, significantly lower LgCL responses were recorded by SOD than catalase. LmCL also showed significantly decreased responses by the addition of SOD and catalase. However, there were no statistical differences in CL responses between SOD and catalase additions. More profound and significant decrease of LmCL responses were recorded by simultaneous addition of SOD and catalase. Sodium azide markedly enhanced LgCL responses, while it significantly inhibited LmCL responses. These results indicate that LgCL and LmCL can be used to measure extracellular $O_2$ production and myeloperoxidase (MPO)-mediated ROS production in fish phagocytes, respectively. Furthermore, LmCL can be used for analyzing intracellular ROS production by simultaneous addition of both SOD and catalase.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.5
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• pp.647-653
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• 2015

A 12-wk feeding trial was conducted to evaluate the essentiality of choline supplementation in diets for parrot fish. Five isonitrogenous and isocaloric diets were supplemented with 0 (as control), 500, 1,000, and 2,000 mg choline per kg diet, and a positive control diet without choline contained 0.3% of 2-amino-2-methyl-1-propanol as choline biosynthesis inhibitor (designated as Con, C500, C1000, C2000 and $Con^+$, respectively). Triplicate groups of fish (body weight, $8.8{\pm}0.01g$) were fed one of the experimental diets at a rate of 4% body weight twice daily. The fish fed $Con^+$ diet revealed significantly lower growth performance and feed utilization efficiency than other fish groups. Supplementation of choline to the basal diet did not significantly influence fish growth. The highest liver lipid content was observed in fish fed the $Con^+$ diet and inversely correlated with liver choline concentration although the differences were not significant. Also, significantly higher liver linoleic, eicosapentaenoic and docosahexaenoic acid contents were found in fish fed the $Con^+$ diet. Innate immune parameters including respiratory burst and myeloperoxidase activities were not significantly affected by dietary choline levels. The findings in this study conclude that choline concentration of approximately $230mgkg^{-1}$ diet meets the requirement of parrot fish.

• Fisheries and Aquatic Sciences
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• v.16 no.1
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• pp.7-14
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• 2013

A 10-week feeding trial was conducted to examine the effects of dietary spirulina and quercetin on growth, innate immunity, disease resistance and dietary antioxidant capacity in the juvenile olive flounder. Triplicate groups of fish (initial body weight, $2.9{\pm}0.01g$) were fed one of isonitrogenous (48% crude protein) and isocaloric (17.4 MJ/kg DM) experimental diets containing 0% spirulina (as a control), 3.4% spirulina, or 6.8% spirulina with or without supplementation of 0.5% quercetin (designated as CON, SP3.4, SP6.8, and SP6.8 + Q, respectively) at a rate of 3% body mass twice daily. Higher dietary antioxidant capacity was found with spirulina supplementation, and the highest value (P < 0.05) was obtained with SP6.8 + Q diet. At the end of the feeding trial, no significant effects were observed on growth performance, body composition and disease resistance against Edwardsiella tarda. Lysozyme activity was significantly increased by spirulina supplementation (P < 0.05), and the highest value was observed in the group fed SP6.8 + Q diet. Also, significantly higher respiratory burst activity (P < 0.05) was found in SP3.4 group. According to the results of this study, dietary supplementation of 3.4% spirulina may enhance innate immunity of olive flounder.

• Journal of fish pathology
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• v.28 no.1
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• pp.53-62
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• 2015

Here, we report the physiological changes and growth in disk abalone, Haliotis discus discus, in relation to dietary supplementation with citrus pomace (CP) 6%, Ecklonia cava residue (ECR) 6%, and CP + ECR (3% + 3%). The composition and nutrient content, survival rate and growth rate were measured 0, 4, 8 and 12 weeks after feeding the supplemented diets of CP and/or ECR. Moreover, the experiment of low salinity stress (25psu) for environmental resistance was examined for a period of 48 hours after feeding the supplemented diets for 12 weeks. The activities of superoxide dismutase (SOD), catalase (CAT), lysozymes, respiratory burst, and phenoloxidase were measured. The moisture content and crude protein condition of the body were increased with the addition of ECR only (P<0.05). We observed higher levels of survival in the experimental group compared with the control group. Moreover, the growth disk abalone that were fed a diet containing ECR was higher compared with the control group. However, the growth of abalone fed a diet containing CP was similar to the control group. With a rearing condition of low salinity stress, survival rate and lysozyme activity were increased in the ECR group compared with the control group. Dietary ECR reduced the level of CAT activity to approximately 30% of the control, however the level of CAT activity in the ECR group was similar to the start level of the previous stress. These results suggest that dietary ECR gives rise to an enhanced immunity in disk abalone, as a result of the decrease in CAT and lysozyme activity in particular. Accordingly, the growth and survival rate were increased by feeding an ECR-supplemented diet in the rearing of disk abalone, Haliotis discus discus.

• Journal of fish pathology
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• v.32 no.1
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• pp.21-28
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• 2019

It has been reported that various anti-oxidant substances stimulate non-specific immune responses in fishes. In this study it was examined whether N-acetylcysteine (NAC), a precusor for anti-oxidant glutathione (GSH) synthesis, can modulate non-specific immune function in Far Eastern catfish Silurus asotus. Immune functions were assessed using the respiratory burst activity monitored by chemiluminescence (CL) responses in isolated leucocyte. NAC stimulated CL responses with doses of 10 or 100 mg/kg, but not with 1 mg/kg after 48 hr injection. It was observed with 10 mg/kg NAC that CL activity continued to elevate from 24 hr through 96 hr post-dosing, and returned to the near preinjection level by 10 days. To understand whether NAC can also activate CL activity in vitro, NAC was directly added to isolated catfish leucocytes. It was observed, however, that NAC can not stimulate CL at reasonable concentration ranges in vitro. As NAC is a precursor for the strong anti-oxidant glutathione (GSH), a putative immune stimulator, it was assessed whether GSH can also stimulate CL responses. Observed results show that GSH activated CL both in vivo and in vitro. The data obtained collectively support the proposition that NAC indirectly stimulates non-specific immune functions in catfish by enhancing GSH biosynthesis, but not by direct action of NAC. Such effects may have beneficial significance in aquaculture for practical utilization.

• Molecules and Cells
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• v.46 no.6
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• pp.329-336
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• 2023

Reactive oxygen species (ROS) serve as secondary messengers that regulate various developmental and signal transduction processes, with ROS primarily generated by NADPH OXIDASEs (referred to as RESPIRATORY BURST OXIDASE HOMOLOGs [RBOHs] in plants). However, the types and locations of ROS produced by RBOHs are different from those expected to mediate intracellular signaling. RBOHs produce O₂^•- rather than H₂O₂ which is relatively long-lived and able to diffuse through membranes, and this production occurs outside the cell instead of in the cytoplasm, where signaling cascades occur. A widely accepted model explaining this discrepancy proposes that RBOH-produced extracellular O₂^•- is converted to H₂O₂ by superoxide dismutase and then imported by aquaporins to reach its cytoplasmic targets. However, this model does not explain how the specificity of ROS targeting is ensured while minimizing unnecessary damage during the bulk translocation of extracellular ROS (eROS). An increasing number of studies have provided clues about eROS action mechanisms, revealing various mechanisms for eROS perception in the apoplast, crosstalk between eROS and reactive nitrogen species, and the contribution of intracellular organelles to cytoplasmic ROS bursts. In this review, we summarize these recent advances, highlight the mechanisms underlying eROS action, and provide an overview of the routes by which eROS-induced changes reach the intracellular space.

• Korean Journal of Microbiology
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• v.49 no.2
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• pp.172-178
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• 2013

The strains were added to the feed in the concentration of $10^3$, $10^5$, and $10^7$ CFU/kg and 2% of fishes were given the feed twice a day (8 AM and 5 PM) for 12 weeks. In result of the nonspecific immune response study to examine Respiratory burst activity, Lysozyme activity and Phagocytosis activity every two weeks until the end of the study, all test samples showed greater activities than control samples and improved immune activity with Bacillus sp. IS-2. The mortality test performed by artificial infection using Streptococcus iniae, a pathogenic bacterium, after the completion of this study also showed over 55% greater survival rate in all test samples. In result of performing PCR using the universal primer to verify that the probiotic stays in the intestines of the fishes, all test samples showed PCR product of 1,465 bp. Based on the above findings, it was concluded that Bacillus sp. IS-2 in the feed improved farmed flatfish's immune system and resistance against diseases as the probiotics. Also, the physiological indicators discovered by this study would be useful for identifying the mechanisms of probiotics.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.3
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• pp.250-256
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• 2009

A 8-week feeding trial was conducted to investigate the effects of dietary supplementation of Kugija (Lycium chinense) on the growth and immunological response in juvenile Korean rockfish (Sebastes schlegeli). Six experimental diets were supplemented with Kugija at 0, 0.1, 0.5 1.0, 3.0 and 5.0% ($K_0,\;K_{0.1},\;Ko_{0.5},\;K_{1.0},\;K_{3.0},\;K_{5.0}$) on a dry-matter basis. After 2 weeks, triplicate groups of 30 fish initially averaging 3.36$\pm$0.2 g (mean$\pm$SD) were randomly distributed into the aquarium and were fed one of the experimental diets for 8 weeks. By the end of the 8-week feeding trial, fish fed the $K_{0.5}$ and $K_{1.0}$ diets exhibited a higher weight gain and specific growth rate than fish fed $K_0$ and $K_{0.1}$ diets (P<0.05). Feed efficiency of fish fed the $K_{0.5}$ diet showed significant higher value than that of fish fed the $K_0$, $K_{3.0}$ and $K_{5.0}$ diets (P<0.05). Hepatosomatic index of fish fed the $K_{0.5}$ diet was significantly higher than that of fish fed the $K_{0.1}$ and $K_{5.0}$ diets (P<0.05). Hematocrit of fish fed the $K_{0.5},\;K_{1.0}$ and $K_{5.0}$ diets was significantly higher than that of fish fed the $K_0$ diet (P<0.05). Glutamic oxaloacetic transaminase of fish fed the $K_{0.5},\;K_{1.0},\;K_{3.0}$ and $K_{5.0}$ diets was significantly lower than in fish fed the other diets (P<0.05). Glutamic pyruvic transaminase activity of fish fed the $K_{3.0}$ diet was significantly lower than those of fish fed the $K_0$ and $K_{0.1}$ diets (P<0.05). However, there was no significant difference in the activity of the transaminase in fish fed the $K_{0.5}\;K_{1.0},\;K_{3.0}$ and $K_{5.0}$ diets. Lysozyme activity of fish fed the $K_{0.5}$ and $K_{1.0}$ diets was significantly higher than that in fish fed the $K_0$ and $K_{0.1}$ diets (P<0.05). Respiratory burst activity of fish fed the $K_{3.0}$ diet was significantly higher than those of fish fed the $K_0$ and $K_{0.1}$ diets (P<0.05). However, there was no significant difference in respiratory burst activity of fish fed the $K_{0.5}\;K_{1.0},\;K_{3.0}$ and $K_{5.0}$ diets. Fish fed Kugija showed lower early mortality than fish fed a Kugija-free diet upon challenge with Vibrio ordalii. The results suggest that feeding of Kugija (Lycium chinense) enhances growth, non-specific immunity and disease resistance in juvenile Korean rockfish.

• Journal of Aquaculture
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• v.21 no.2
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• pp.82-88
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• 2008

A 12-week feeding trial was conducted to investigate the effects of dietary supplementation of probiotics as a feed additive for juvenile Korean rockfish Sebastes schlegeli. Four experimental diets supplemented with no probiotic(Control), Bacillus polyfermenticus(BP), Bacillus licheniformis(BL) or Bacillus polyfermenticus plus Saccharomyces cerevisiae(BP+SC) at $1.0{\times}10^7$ CFU/kg diet as a dry-mater(DM) basis were prepared by mixing with a basal diet. After 12 weeks of the feeding trial, fish fed BP+SC diet showed significantly higher weight gain(WG), feed efficiency(FE), specific growth rate(SGR) and protein efficiency ratio(PER) than those of fish fed control diet(P<0.05), however there were no significant differences in WG, FE, SGR and PER among fish fed the BP, BL and BP+SC diets. Fish fed BP and BP+SC diets showed significantly higher condition factor(CF) than that of fish fed control and BL diets. Fish fed BP, BL, BP+SC diets showed significantly higher hepatosomatic index(HSI) than that of fish fed control diet, however there was no significant difference in HSI among fish fed BP, BL and BP+SC diets. Fish fed BP+SC diet showed significantly lower serum glucose than that of fish fed control diet, however there was no significant difference in serum glucose among fish fed BP, BL and BP+SC diets. Fish fed BP+SC diet showed significantly higher respiratory burst activity(NBT assay) than that of the fish fed control and BL diets, however there was no significant difference in NBT assay between fish fed BP and BP+SC diets. Fish fed BP and BL diets showed significantly higher lysozyme activity than that of the fish fed control diet, however there was no significant difference in lysozyme activity among fish fed BP, BL and BP+SC diets. Fish fed BP and BP+SC diets showed significantly lower cumulative mortality than that of the fish fed control diet, however there was no significant difference in cumulative mortality among fish fed BP, BL and BP+SC diets after the challenge test. From these results, dietary B. polyfermenticus, B. licheniformis and B. polyfermenticus plus S. cerevisiae supplementation in juvenile Korean rockfish diet could enhance growth performances, non-speicific immunities and a higher resistance against the specific pathogen.

• Tuberculosis and Respiratory Diseases
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• v.61 no.3
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• pp.256-264
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• 2006

Background: An acute lung injury(ALI) is characterized by the recruitment, activation, and apoptosis of inflammatory cells, numerous products released by inflammatory cells such as reactive oxygen species, inflammatory mediators, and a variety of proteolytic enzymes. It was reported that bacterial infections in diabetics showed impaired PMN functions such as reduced PMN respiratory burst and decreased microbicidal activity in inflamed tissue. However, the effect of the proteinase - inhibitor (MMP-9 vs TIMP-1) in ALI in diabetics is unclear. This study evaluated the differences in the expression of MMP-9 and TIMP-1 after the stimulation of endotoxin in a rat model. Methods: Six-week-old male Sprague-Dawley rats were classified into normal, DM, LPS and DM+LPS groups. The peripheral blood, BAL fluids, and lung tissues were obtained from individual rats. The MMP-9 activity was measured by gelatin zymography and the TIMP-1 level was measured by Western blotting. Results: The total BAL cells of the DM-LPS groups were significantly lower than the LPS groups (p < 0.01). The MMP-9 activities in the serum were higher in the DM+LPS groups than in the other groups. The MMP-9 activities in the BAL fluids were significantly higher in the DM+LPS group than in the normal and diabetic rats (p < 0.05). TIMP-1 expressions in the BAL fluids were significantly lower in the DM+LPS group than other groups (p < 0.05). The ratio between MMP-9 and TIMP-1 in the BAL fluids was significantly higher in the DM+LPS groups (p < 0.05). Conclusion: In ALI in diabetics the higher MMP-9 activity and lower TIMP-1 level are believed to prolonged and intensify the course of inflammation.