• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.93-100
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• 2020

A bacterial strain inhibiting the growth of Vibrio anguillarum, the causative agent of vibriosis, was isolated from fish intestines. The isolated strain HS36 was identified as Aerococcus urinaeequi based on the characteristics of the genus according to Bergey's Manual of Systematic Bacteriology and by 16S rRNA sequencing. The growth rate and antibacterial activity of strain HS36 in shaking culture were higher than those in static culture, while the optimal pH and temperature for antibacterial activity were 7.0 and 30℃, respectively. The active antibacterial substance was purified from a culture broth of A. urinaeequi HS36 by Sephadex G-75 gel chromatography, Sephadex G-25 gel chromatography, and reverse-phase high-performance liquid chromatography. Its molecular weight, as estimated by Tricine SDS-polyacrylamide gel electrophoresis, was approximately 1,000 Da. The antibacterial substance produced by strain HS36 was stable after incubation for 1 h at 100℃. Although its antibacterial activity was optimal at pH 6-8, activity was retained at a pH range from 2 to 11. The purified antibacterial substance was inactivated by proteinase K, papain, and β-amylase treatment. The newly purified antibacterial substance, classified as a class II bacteriocin, inhibited the growth of Klebsiella pneumoniae, Salmonella enterica, and Vibrio alginolyticus.

• Microbiology and Biotechnology Letters
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• v.31 no.2
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• pp.129-134
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• 2003

The purpose of the present study was to screen the effective of lactic acid bacteria (LAB) as probiotics, which are able to protect aquacultural fish pathogenic bacteria, and investigate their characterization. Twenty strains of lactic acid bacteria were isolated from fish intestine, fermented fish foods and kimchis. These bacteria were screened for antagonistic activity against fish pathogenic bacteria. Seven tested LAB strains were able to inhibit the fish pathogenic bacteria, including Vibrio anguillarum, Edwardsiella tarda, and Streptococcus sp.. Of the probiotic candidates, BK19 strain isolated from fermented pollack viscera indicated the largest inhibition activity. Moreover, this strain showed a resistance over low pH and antibiotic agents. Therefore this probiotic candidate BK19 was finally selected and identified as a probiotic strain. This particular probiotic bacteria was identified as Lactobacillus sakei BK19 by biochemical characteristics and 165 rRNA PCR amplification.

• Journal of Life Science
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• v.18 no.6
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• pp.789-795
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• 2008

To study the possible use of probiotics in fish farming, we evaluated antagonism of antibacterial strain Bacillus amyloliquefaciens H41 against the fish pathogenic bacterium Vibrio anguillarum NCMB1. The purification of growth inhibition factor produced by B. amyloliquefaciens H41 was achieved by obtaining supernatant of this bacterium. The growth inhibition factor was purified to homogeneity by 70% ammonium sulfate precipitation, DEAE-sephadex A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography, and sephadex G-50 gel filtration column chromatography with 40.8 fold of purification and 2.9% yield. The molecular weight of the purified growth inhibition factor was 48 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH and temperature for the growth inhibition factor were pH 7.5 and $30^{\circ}C$, respectively. The activity of growth inhibition factor was enhanced slightly by some metal ions, such as $Mg^{+2}$, $Mn^{+2}$, but was inhibited by the addition of $Co^{+2}$, $Hg^{+2}$, $Zn^{+2}$ and $Ag^{+2}$. NaCl stability of the growth inhibition factor was observed with 50% residual activity at 3% NaCl concentration. Toxicity test showed that the purified B. amyloliquefaciens H41 growth inhibition factor did not affect the live of Japanese flounder (Paralichthys olivaceus) and the effectiveness was 78% of residual lethality compared to commercial antibacterial agents.

• Journal of fish pathology
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• v.13 no.1
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• pp.45-51
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• 2000

Bacterial diseases with mixed infection have recently occurred at land-based flounder farms in Korea. Thus, single antibacterial is not effective for therapy of mixed bacterial diseases of fish because of their different causative bacteria. The purpose of the present study was to obtain basic data for positive usefulness of a combination of antibacterials used for synergism to mixed bacterial diseases of fish. Snergistic interaction in combination of antibacterials was determined by in vitro antimicrobial activity against selected fish-pathogenic bacteria, Vibrio anguillarum, Edwardsiella tarda, Streptococcus sp., Staphylococcus epidermidis, on the basis of Checkerboard assay using fractional inhibitory concentration (FIC) indices. Synergistic interactions were observed in combinations of (oxytetracycline HCL+lincomycin), (tetracycline HCL+florfenicol), (oxytetracycline HCL+florfenicol) against V. anguillarum, (sodium nifurstyrenate+florfenicol), (tetracycline HCL+florfenicol), (sodium nifurstyrenate+oxolinic acid), (oxytetracycline HCL+florfenicol) against E. tarda, (ciprofloxacine+oleandomycin), (oxytetracycline HCL+oleandomycin), (tetracycline HCL+oleandomycin), (oxytetracycline HCL+lincomycin), (oxytetracycline HCL+spiramycin), (oxytetracycline HCL+erythromycin), (doxycycline HCL+oleandomycin), (tetracycline HCL+spiramycin) against Streptococcus sp., and (ciprofloxacine+erythromycin), (florfenicol+erythromycin), (doxycycline HCL+oleandomycin), (ciprofloxacine+oleandomycin) against S. epidermidis.

• Journal of Life Science
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• v.17 no.10
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• pp.1434-1440
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• 2007

We have investigated the effects of mushroom mycelium (Phellinus linteus, Coriolus versicolor) mixed cultural extract on the immune responses of olive flounder, Paralichthys olivaceus. The mixed culture extracts were evaluated for the growth, hematology, lysozyme activity, leukocyte phagocytic activity, and disease resistance against Vibrio anguillarum. In the effect of the growth, the body weight and length gain in the group, which fed with mushroom mycelium mixed cultural extract, were 52 g and 3.4 cm higher than that in the control, respectively. For the hematology, the administration of mushroom mycelium mixed cultural extract resulted in increase of glucose. However, there was no distinct differences in GOT (glutamic oxaloacetic transaminase), GPT (glutamlc pyruvic transamlnase), TG, TP, and LDH (lactate dehydronase) among each group. The activities of lysozyme were 80% higher in the experimental groups than in the control. The activities of leucocyte were 66% higher in the experimental groups than in the control. Although lysozyme activity and leucocyte activity showed somewhat decrease after 12 weeks, these activities were still higher than in the control. The cumulative mortality (%) after an artificial challenge with $7{\times}10^8\;CFU$ of Vibrio anguillarum per fish was 25% higher in the experimental groups than the control.

• Journal of fish pathology
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• v.22 no.2
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• pp.115-124
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• 2009

Photobacterium damselae subsp. damselae and 4 Vibrio spp, V. anguillarum, V. splendidus, V. harveyi and V. ordalii, were isolated from the diseased olive flounders, Paralichthys olivaceus. The pathogenicity of the isolates were compared to mortality, blood biochemical contents, such as alanineaminotransferase(ALT), aspatate aminotransferase(AST), and cortisol level, and non-specific immune responses, nitroblue tetrazolium (NBT) reduction of macrophages and lysozyme activities of serum. The mortalities and levels of ALT, AST and cortisol of fishes infected with P. damselae were higher than those of others but significantly low in non-specific immune responses, NBT and lysozyme activities. These results suggest that P. damsela might produce damselysin having high hemolytic and phospholipase activities, correlated with the pathogenicity. P. damsela could also make an obstruction of internal organs, following increasing in the level of ALT and AST, and depression in host immunity caused by induced high levels of cortisol.

• Journal of Life Science
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• v.27 no.10
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• pp.1161-1167
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• 2017

In order to find a new antimicrobial bacterium, we performed screening for antimicrobial activity of bacteria isolated from the eggs of a sea hare. The newly identified strain was designated as Phaeobacter inhibens KJ-2, based on the biochemical characterization and 16S rRNA gene sequence analysis. A colony of P. inhibens KJ-2 showed a circular and ruler-like smooth form at the edge, and a brown color. However, when maintained with a longer incubation time, its coloring was transformed into dark brown. From the result of SEM, P. inhibens KJ-2 is a bacillus which has a length of $0.8{\sim}1.0{\mu}m$ and a width of $0.4{\sim}0.6{\mu}m$. The optimal growth and antimicrobial activity were observed by shaking the culture for 24 hr at $20^{\circ}C$, which showed potent activity against pathogenic bacteria including Vibrio logei, Vibrio campbellii, Vibrio mimicus, Vibrio vulnificus, and Vibrio salmonicida. The antimicrobial activity was proportional to the amount of produced acylated homoserine lactones (AHLs). Therefore, we suggest that production of antimicrobial materials from P. inhibens KJ-2 is regulated by Quorum sensing (QS).

• Journal of fish pathology
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• v.20 no.1
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• pp.71-81
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• 2007

Photobacterium damselae subsp. damselae and 4 Vibrio spp.(V. anguillarum, V. splendidus, V. harveyi and V. ordalii) were isolated from the diseased olive flounders, Paralichthys olivaceus. The isolates were tested on the pathogenicity in vitro. The properties of extracellular products(ECPs) were investigated with enzymatic activities, hemolytic activities toward the sheep and olive flounder erythrocytes, and cytotoxicity activities on the cell-line. And potential signal transduction pathways of the bacterial internalization were detected by using signal transduction inhibitors. P. damselae was high in phospholipase activity, hemolytic activity to olive flounder erythrocytes and cytotoxicity activity. And P. damselae had diversified internalizing pathways as compared to isolated vibrios. Therefore, these activities may be related with pathogenicity of P. damselae.

• Proceedings of the Korean Society of Life Science Conference
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• 2003.10a
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• pp.103-103
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• 2003

• Journal of Aquaculture
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• v.19 no.4
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• pp.231-235
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• 2006

The effects of mycelium cultural extract supplemented diet, hematology and disease resistance against Vibrio anguillarum in juvenile flounder, Paralichthys olivaceus were evaluated. Fish were fed the Phellinus linteus with Coriolus militaris versicolor mixed mycelium cultural extract supplemented diet, Phellinus linteus mycelium cultural extract supplemented diet and Coriolus militaris tmycelium cultural extract supplemented diet a commercial diet for 12 week. The body weight and length gain from the fish fed on daily the phellinus with coriolus versicolor mixed mycelium cultural extract supplemented diet, phellinus linteus mycelium cultural extract supplemented diet and Coriolus versicolor mycelium cultural extract supplemented diet of each mycelium cultural extract were higher than the control and the Glutamic oxalacetic transaminase (GPT) were of lower than the control. The relative precent survival rate (RPS) after an artificial challenge with $7{\times}10^5\;CFU$ of Vibrio anguillarum per fish was higher than the control.