• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.6
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• pp.823-829
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• 2016

Yacon (YC), ginger (GG) and blueberry (BB) were added to the diet of juvenile rockfish Sebastes schlegeli to investigate the effects of these substances on growth, feed utilization and resistance to Streptococcus iniae compared to feed additive ethoxyquin. A total of 480 fish were distributed in 12 tanks and four experimental diets [control (Con), with 0.01% ethoxyquin, and 1% YC, GG and BB diets] were provided in triplicate. After an 8-week feeding trial, 20 fish from each tank were infected with Streptococcus iniae. Fish fed the BB diet had higher weight gain than fish fed the GG and Con diets. Feed efficiency of fish fed the GG and YC diets was higher than that of fish fed the BB and Con diets. The protein efficiency ratio of fish fed the GG diet was higher than that of fish fed any other diet. The cumulative mortality 54 h after infection of fish fed the Con diet was higher than that of fish fed any other diet. These results indicate that YC, GG, and BB are effective dietary additive, not only improving growth performance and feed utilization but also lowering the mortality of juvenile rockfish exposed to S. iniae.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.5
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• pp.678-682
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• 2016

Antibacterial activity of 80% methanol extract from 13 commercial herb medicines was measured against fish pathogens Streptococcus iniae, causing several diseases in various kind of fish. Siegesbeckia pubescens showed the strongest antibacterial activity against S. iniae. Methanol extract of S. pubescens was further extracted using several organic solvents having different polarity. Extract from n-hexane and ethyl acetate fraction showed strong activity. Minimal inhibitory concentration, MIC of S. pubescens extract was measured and resulted showing 8 μg/mL with n-hexane fraction and 32 μg/mL with ethyl acetate fraction against S. iniae. The growth of S. iniae was fully inhibited by adding 50 μg/mL (final concentration) of n-hexane or ethyl acetate fraction in the liquid media. It is needed that, from these results, purification and isolation of responsible active compound(s) of these activities and further study on the synergy effect using combination with commercial antibiotics against fish pathogenic bacteria.

• Natural Product Sciences
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• v.24 no.4
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• pp.225-228
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• 2018

By activity-guided fractionation, gliotoxin was isolated as an antibacterial metabolite of the fungus Penicillium decumbens which was derived from the jellyfish Nemopilema nomurai. Gliotoxin was further evaluated for antibacterial activity against several piscine and human MDR (multidrug resistance) pathogens. Gliotoxin showed significant antibacterial activity against Gram-positive piscine pathogens such as Streptococcus iniae FP5228, Streptococcus iniae FP3187, Streptococcus parauberis FP3287, Streptococcus parauberis SPOF3K, S. parauberis KSP28, and Lactococcus garvieae FP5245. Gliotoxin showed strong activity especially against S. parauberis SPOF3K and S. iniae FP5228, which are resistant to oxytetracycline. It is noteworthy that gliotoxin effectively suppressed streptococci which are the major pathogens for piscine infection and mortality in aquaculture industry. Gliotoxin also showed strong antibacterial activity against multidrug- resistant human pathogens (MDR) including Enterococcus faecium 5270 and MRSA (methicillin-resistant Staphylococcus aureus) 3089.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.1
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• pp.18-22
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• 2014

The antibacterial activities of methanol extracts of 19 commercial herbal medicines were measured against the fish pathogens Vibrio ichthyoenteri and Streptococcus iniae, which cause several fish diseases. Rhus javanica showed the strongest antibacterial activity against V. ichthyoenteri and S. iniae. The methanol extract of R. javanica was extracted further using several organic solvents with different polarities. The extract from the ethyl acetate fraction showed strong activity against both fish pathogens. The minimum inhibitory concentration (MIC) of the R. javanica extract was $32{\mu}g/mL$ for V. ichthyoenteri and $128{\mu}g/mL$ for S. iniae. Further purification and isolation of the active compound (s) responsible for these activities and further study of the synergistic effect using combinations of antibiotics against pathogenic bacteria are needed.

• Korean Journal of Veterinary Research
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• v.47 no.3
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• pp.291-298
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• 2007

Olive flounder, Paralichthys olivaceus is one of the most important cultured fish in Korea, its farming has been negatively impacted by viral, bacterial and parasitic diseases. Streptococcal infection was considered as a serious problem because of significant economic losses in olive flounder farm industry. The development and evaluation of vaccine for protection against infection by this agent were required. We evaluated the safety and efficacy of ${\beta}$-hemolytic Streptococcus (S.) iniae vaccine on olive flounder Three hundreds of flounders (weight $119.8{\pm}20.7g$, body length $22.6{\pm}1.4cm$) were reared in 0.5 tons aquaria in land-marine tank system. Seawater was provided from the sea of Inchon in Korea, and water temperature was set to $22^{\circ}C$ and $25^{\circ}C$ in the vaccination and challenge test, respectively. We used the formalin-inactivated ${\beta}$-hemolytic S. iniae (F2K) vaccine (M VAC INIAE; Kyoritsu seiyaku, Japan) originated in Japan. The vaccine was intraperitoneally administered to fish. Both of vaccinated group and control group were challenged with intraperitoneally injection by virulent S. iniae SI-36 isolates with $1.0{\times}10^7CFU/fish$ at 3 weeks after vaccination. Difference on mortality of control and vaccinated group (90.0 and 15.0%, 76.5 and 8.0% respectively) at two trials were found significant (p<0.05), and relative percent survival were 83.4% and 89.5%, respectively. The dead fishes were showed dark pigmentation of skin, abdominal extension, hemorrhagic ascites, and liver necrosis, and isolated the S. iniae strain from ascites, liver and kidney. We confirmed the safety and efficacy of ${\beta}$-hemolytic S. iniae vaccine by determinations of the optimal management condition and artificial challenge test in olive flounder.

• Journal of fish pathology
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• v.33 no.2
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• pp.119-125
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• 2020

During the period from 2003 to 2020, a total of 470 Streptococcus species were isolated from farmed flounder in Jeju. Out of 470 isolates, 92 (19.6%) isolates and 378 isolates (80.4%) were identified as Streptococcus iniae and S. parauberis, respectively by multiplex PCR assay. During that period, the percentage of S. iniae decreased from 56.9% in 2003 to 0.0% in 2020 whereas that of S. parauberis increased from 43.1% in 2003 to 100% in 2020. In the PCR assay for serotyping, the isolated S. parauberis showed 3 subserotypes, Ia (34.9%), Ib/Ic (46.3%) and II (18.8%). In 2003 and 2004, serotype II was dominant at 59.1% and 50.0% of isolation rates, however between 2005 and 2009, subserotype Ib/Ic was dominant (57.6%, 86.0%, 84.6%, 57.9%, and 83.3%). After 2010, except for 2015, subserotype Ia was the most dominant one. In the last 3 years (2018 to 2020), subserotype Ia was most abundant (70%), followed by subserotype Ib/Ic (16-30%). Serotype II was not isolated in 2018, but in 2019 and 2020, it showed an increased tendency to 3.4% and 16.7%, respectively. It is believed that continuous monitoring is necessary for research on counter-measures against streptococcosis of flounder.

• Journal of fish pathology
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• v.18 no.2
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• pp.167-178
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• 2005

Streptococcus iniae is one of the major bacterial pathogens of cultured olive flounder, Paralichthys olivaceus in Korea. This study was carried out to investigate the interaction between morphological characteristics and immune response such as phagocytosis and serum killing activity. They were differentiated phenotypically into two groups, the viscous colonies and the non-viscous colonies, but the strains in both groups were similar in physiological and biochemical characteristics. Electron microscopic examination of the viscous form revealed thick, electron-dense exopolysaccharide materials surrounding polycationic ferritin-stained cells, while the exopolysaccharide material was absent around the non-viscous form of S. iniae. The viscous type strains were disclosed more virulent than those of non-viscous type for olive flounder. The viscous strains were resistant to normal serum killing activity, while the non-viscous strains were susceptible to bactericidal activity of normal serum from olive flounder. The viscous strains were more resistant to opsonophagocytosis and decreased the chemiluminescence response of head kidney macrophages of olive flounder. All of the tested non-viscous S. iniae strains were efficiently destroyed by the macrophage after phagocytosis. Both the viscous and the non-viscous strains invaded and replicated in cultured fish cell line (CHSE-2l4). This cellular invasion may contribute to the pathogenesis of invasive S. iniae infection.

• Journal of Veterinary Clinics
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• v.26 no.3
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• pp.226-230
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• 2009

We evaluated the efficacy of $\beta$-hemolytic Streptococcus(S.) iniae vaccine on cultured olive flounder. Three hundred flounders(weight $50{\pm}5$ g) were obtained from two farm at Wando and Taean in the southern and western coast of Korea at May and June 2007, respectively. Twenty of flounders moved in 0.5 tons aquaria in land-marine tank system of National Veterinary Research and Quarantine Service. Seawater was transported from the sea of Inchon in western Korea, and water temperature maintained to $22^{\circ}C$ and $25^{\circ}C$ during the vaccination and challenge test, respectively. We used the formalin-inactivated $\beta$-hemolytic S. iniae vaccine produced by domestic manufacturers. The vaccine was intraperitoneally administered to fish. The vaccinated and control group were challenged with intraperitoneal injection by virulent S. iniae SI-36 isolates with $5.0{\times}10^8$ CFU/fish at 3 weeks after vaccination. We evaluated the vaccine efficacy by calculating numbers of dead fish, and observing of clinical signs, exterior and gross lesions, and examining bacteria isolation and identification. Thirty-four(25.2%) of 135 control and vaccinated group fish were dead with serious anemia, abdominal extension, and hernia of intestine during 3 weeks post vaccination. We isolated Neoheterobothrium hirame from the buccal cavity and Edwardsiella tarda from kidney of dead and diseased fish. When infected fish with these agents were challenged with S. iniae SI-36 isolates, the cumulative mortality of control and vaccinated group were 86.7, and 46.7%, respectively. However, significant differences(p<0.05) were observed on cumulative mortality between control(20.0%) and vaccinated group(95.0%) at second trials with 40 healthy, and relative percent survival(RPS) was 78.0%. We confirmed that the efficacy of $\beta$-hemolytic S. iniae vaccine on olive flounder were impacted by health condition such as bacterial and parasitic diseases.

• Journal of fish pathology
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• v.36 no.2
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• pp.277-286
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• 2023

Korean rockfish, Sebastes schlegelii, is a representative bony fish that belongs to the family Scorpaenidae and the order Scorpaeniformes. It has high ecological and economic value and is widely cultivated in many East Asian countries, including South Korea, Japan and China. One of streptococci, Streptococcus iniae, is Gram-positive cocci with a negative reaction for catalase and oxidase. The Korean rockfish shows clinical signs when infected with S. iniae, such as body darkening, bleeding, enlarged kidneys, blurred eyes, abdominal distension, etc., ultimately leading to death. The Korean rockfish causes significant economic losses every year in South Korea due to streptococcosis. In this study, we identified bacteria from the fish using polymerase chain reaction and conducted analyses of hemolytic activity and biochemical tests using API 20 STREP and API ZYM systems. Results of confirming the hemolytic activity (n=4) observed in alpha-type hemolysis (25%), beta-type hemol- ysis (50%), and gamma-type hemolysis (25%) of isolates. The biochemical test results exhibited sig- nificant variation among S. iniae. Additionally, we performed intraperitoneal injection with S. iniae in the fish and analyzed the phylogenetic tree using housekeeping genes of S. iniae, including cpsD, arcC, glnA, groEL, gyrB, mutS, pheT, prkC, rpoB, and tkt, via multilocus sequence typing (MLST). The lethal dose (LD₅₀) showed strong pathogenicity, such as 3.34 × 10 colony-forming unit (CFU)/ml for 23FBStr0601 strain and 7.16 × 10 CFU/ml for 23FBStr0602 strain. 23FBStr0603 strain showed relatively low pathogenicity at 1.73 × 10⁵ CFU/ml. The strains 23FBStr0601 and 23FBStr0602, which showed strong pathogenicity, clustered into one monophyletic group. The 23FBStr0603 strain showed weak pathogenicity and formed a monophyletic group with KCTC 3657.

• Journal of Life Science
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• v.24 no.8
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• pp.880-886
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• 2014

In this study, we isolate and identify bacteria from seawater collected from Jeju coast, to evaluate the antimicrobial activity against the fish pathogenic bacteria. 14 bacterial strains were isolated and identified using physiological, biochemical and molecular tools. Antibacterial activity of all the 14 isolates were screened against four major fish pathogens namely, two Gram-positive: Streptococcus iniae, Streptococcus parauberis and two Gram-negative: Vibrio anguillarum, Edwardsiella tarda. Results revealed that among the 14 isolates, MK-11 was found to have antibacterial activity against S. iniae, S. parauberis, V. anguillarum Particularly, S. iniae was susceptibility with the MIC value of $250{\mu}g/ml$. The biochemical and physio-chemical results reveal that MK-11 had the sugar-alcohol disassemble ability of the D-sorbitol and D-mannitol. Also the utilization of the yeast extract, sorbitol and di-potassium phosphate were noted to be high. The optimum culture condition such as pH and temperature was recorded as pH 6.0, $25^{\circ}C$ and along with 1% NaCl which differs from the previous reports particularly in nutrient resolutions. As results of the analysis of 16S rDNA sequences, MK-11 show the high similarity with Paenibacillus polymyxa, P. jamilae, P. brasilensis 99.78%, 99.43%, 99.39%, repectively. Hence, in the present study, the isolated Paemibacillus sp. MK-11 from Jeju seawater possesses the antibacterial activity against fish pathogens and it could be used as a new antibiotic agents against the gram positive fish pathogens.