• Korean Journal of Environmental Biology
• /
• v.32 no.4
• /
• pp.382-388
• /
• 2014

Shortfin eel (Anguilla bicolor pacifica) is a species of commercial importance and its production is greatly affected due to the infection by Heterosporis anguillarum. In this study, we evaluated the effect of H. anguillarum infection on the growth of Shortfin eel. A disease that trunk muscle of cultured shortfin eel, Anguilla bicolor pacifica, were irregular and resulted in death, breakout of the commercial eel culture farm. We observed that the trunk muscle of infected eels were irregular and represented white or yellowish externally. Histopathologically, a great numbers of large or small spores and sporophorocysts were also observed in degenerated muscle layer. The cloning of specific gene of H. anguillarum, encoding small subunit ribosomal RNA (SSU-rRNA) was amplified by the polymerase chain reaction(PCR) from the muscle lesion of diseased eel. The size of clone gene is well matched with the size of small subunit ribosomal RNA of H. anguillarum and thus confirming the infection by H. anguillarum.

• Journal of Life Science
• /
• v.16 no.4
• /
• pp.605-611
• /
• 2006

To investigate the possible use of probiont in fish farming industry, a bacterium with inhibitory effect against Vibrio anguillarum was isolated from gastrointestinal tract of the marine fish of yellow tail. It was identified to be Bacillus amyloliquefaciens H41 based on biochemical and physiological characterization. The optimal growth conditions of the isolated strain were 1% peptone, 1.5% yeast extract, 1% sucrose, 0.5% NaCl, 0.05% $MgSO_4{\cdot}7H_20$, pH 7.0-8.0, and 20 hr of incubation between $28-35^{\circ}C$ under aeration. The culture supernatant of the isolated strain showed inhibition activity against V. anguillarum. Inhibition activity was cleared by forming a clear zone by a paper-disk method. The maximal production of growth inhibition factor was induced by cultivation under 1% peptone, 1.5% yeast extract, 1% sucrose, 1% NaCl, 0.05% $MgSO_4{\cdot}7H_20$, pH 7.5 and at $35^{\circ}C$. The highest growth inhibition factor production was observed after 16-24 hr cultivation under aeration. The culture supernatant of the isolated strain showed inhibition activity whereas no inhibition activity was shown from the standard B. amyloliquefaciens KCTC 1724 strain. The growth inhibition affected only against V. anguillarum among other pathogenic Vibrios tested here.

• Korean Journal of Veterinary Research
• /
• v.47 no.3
• /
• pp.303-308
• /
• 2007

The high mortality (65.5%) of young olive flounder (Paralichthys olivaceus) showing skin ulcers occurred in fish obtained from a commercial cultured farm at Donghae on the eastern coast of Korea at March 2006. The dorsal skin showed ulcer lesions and large amount of mucus. We isolated Ichthyobodo sp. from the gill, and Vibrio anguillarum from the liver and kidney of dead and diseased fish. All of the fish infected with the V anguillarum and Ichthyobodo sp. were treated with 100 ppm formalin for 1 h, and treated with oxolinic acid for 4 h at $22^{\circ}C$, respectively. The formalin treated group was observed the low mortality (2.9%) when compared with high mortality of non-treated (82.9%), and of antibiotic treated group (85.9%) against mixed infections by Vibrio anguillarum and Ichthyobodo sp. organisms. We confirmed complete elimination against mixed infections of Vibrio anguillarum and Ichthyobodo sp. with 100 ppm formalin treatment.

• Journal of Life Science
• /
• v.8 no.5
• /
• pp.598-603
• /
• 1998

A marine microbe, Vibrio anguillarum was isolated from fish and studied for its concerning pathogenic substance of hemolysin. Purification of hemolysin was achieved by the procedure of ammonium sulfate precipitation from cul-ture filtrate, DEAE-cellulose chromatography, and G-200 gel filtration with 36 fold of purification and 2.3% yield. The molecular weight of the purified hemolysin was 38,000 dalton by SDS-PAGE. The purified hemolysin was stable at pH 6-9, below 45$^{\circ}C$, and up to 1% of NaCl, respectively. $Ca^{2+}, Cu^{2+}, Zn^{2+}, Fe^{2+}$ inhibited the hemolytic activity whereas EDTA and $Mg^{2+}$ did not.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.49 no.3
• /
• pp.359-366
• /
• 2016

The health of Alaska pollock Theragra chalcogramma seedlings was monitored during February and April 2015. After microscopic examination for parasites, 20 samples sets were made by pooling 50 individuals for each sample set. Then, they were homogenized and examined for viral and bacterial pathogens. No parasites or viruses were detected using either microscopy or PCR. Colonies suspected of belonging to the genus Vibrio were isolated from Tryptic Soya Agar and Thiosulfate Citrate Bile Salts Sucrose Agar plate incubations, and identified as Vibrio anguillarum based on biochemical and physiological examinations and PCR amplification of the 16S rDNA, recA, and pyrH genes. Although there was no mortality during the sampling period, 65.0% (13/20) of the pooled samples were PCR-positive for V. anguillarum. To prevent possible outbreaks, the pathogenic potential of V. anguillarum should be investigated in the future.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.8
• /
• pp.1457-1463
• /
• 2016

Vibrio anguillarum, a devastating pathogen causing vibriosis among marine fish, is prevailing in worldwide fishery industries and accounts for grievous economic losses. Therefore, a rapid on-site detection and diagnostic technique for this pathogen is in urgent need. In this study, two mouse monoclonal antibodies (MAbs) against V. anguillarum, 6B3-C5 and 8G3-B5, were generated by using hybridoma technology and their isotypes were characterized. MAb 6B3-C5 was chosen as the detector antibody and conjugated with quantum dots. Based on MAb 6B3-C5 labeled with quantum dots, a modified dot blot assay was developed for the on-site determination of V. anguillarum. It was found that the method had no cross-reactivity with other than V. anguillarum bacteria. The detection limit (LOD) for V. anguillarum was 1 × 10³ CFU/ml in cultured bacterial suspension samples, which was a 100-fold higher sensitivity than the reported colloidal gold immunochromatographic test strip. When V. anguillarum was mixed with turbot tissue homogenates, the LOD was 1 × 10³ CFU/ml, suggesting that tissue homogenates did not influence the detection capabilities. Preenrichment with the tissue homogenates for 12 h could raise the LOD up to 1 × 10² CFU/ml, confirming the reliability of the method.

• Journal of Life Science
• /
• v.18 no.6
• /
• pp.789-795
• /
• 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
• /
• v.18 no.2
• /
• pp.147-156
• /
• 2005

To obtain antimicrobial algae against fish pathogenic bacteria, we screened 80% methanolic extracts of 30 algae using fish pathogenic bacteria, Staphylococcus sp., Streptococcus sp., Edwardsiella tarda and Vibrio anguillarum. Among them, Corallina officinalis, Dumontia simplex, Gloipeltis furcata, Grateloupia lanceolata and Grateloupia turuturu were effective for growth inhibition of a Gram-positive bacterium, Staphylococcus sp.. Sargassum thunbergii and Polysiphonia morrowii exhibited significant inhibitory effects against the growth of Gram-negative bacteria, both E. tarda and V. anguillarum. Moreover, antimicrobial activity-guided fractionation for P. morrowii extract yielded significantly active 90% methanolic fraction. This fraction significantly inhibited the growth of E. tarda exhibiting a MIC of 1 mg/ml. In addition, its antimicrobial activity was stable under various pH conditions.

• Fisheries and Aquatic Sciences
• /
• v.16 no.4
• /
• pp.267-272
• /
• 2013

Chicken-type lysozyme (c-lysozyme) is present in shrimp and is active against some bacteria. To further understand the regulation of c-lysozyme in the fleshy shrimp Fenneropenaeus chinensis, we determined the tissue-specific gene expression of c-lysozyme and the time-course of mRNA expression in response to Vibrio anguillarum and lipopolysaccharide (LPS) injection by quantitative reverse real-time polymerase chain reaction. The results showed that c-lysozyme was expressed in all tissues tested, including gill, eyestalk, eye, hemocytes, hepatopancreas, intestine, heart, and pleopod. It was most highly expressed in the intestine followed by the eyestalk, gill, hemocytes and hepatopancreas. The mRNA expression level began to decline in a short time after V. anguillarum challenge and was then upregulated by two fold or more at 24 h post injection (hpi) compared to that at 0 h. Expression was suppressed shortly after LPS injection and began to increase with higher levels of 5.8-, 5.2- and 8.4-fold at 24, 48, and 72 hpi, respectively. Higher expression was sustained and showed a gradual increasing trend until the end of the experiment (72 hpi). These results increase our understanding of the regulation of defense mechanisms and facilitate an evaluation of the effects of probiotics or immunostimulants in shrimp culture.

• Korean Journal of Microbiology
• /
• v.39 no.1
• /
• pp.56-61
• /
• 2003

The purpose of the present study was to screen the effective of lactic acid bacteria (LAB), as probiotics which are able to protect bacterial fish diseases and investigate their characteristics. 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 which from fermented pollack viscera indicated the largest inhibition activity. This particular probiotic bacteria was identified and named as Lactobacillus sakei BK19. In the scanning electron microscope observation, L. sakei BK19 supernatant treated V.anguillarum cell wall had been destroyed incubate after 3 hr.