• Korean Journal of Microbiology
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• v.41 no.2
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• pp.117-124
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• 2005

Two bacterial isolates obtained from rotifer and diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization. In an attempt to develop rapid PCR method for the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. Analysis of the ISR sequences showed that V. ichthyoenteri contains one type of polymorphic ISRs. The size of ISRs was 348 bp length and did not contain tRNA genes. Mutiple alignment of representative sequences from different V. species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of V. ichthyoenteri. The specificity of the primer was examined using genomic DNA prepared from 19 different V. species, isolated 18group Vibrio species and most similar sequence of other known Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

• Journal of fish pathology
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• v.17 no.3
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• pp.159-169
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• 2004

Bacterial wihte enteritis ocurred by infection of V. ichthyoenteri is a devastating disease in olive flounder (Paralichthys olivaceus) hatcheries in Korea. Since white enteritis has been a problem in aquqtic industries, necessity of a rapid detection method is increased. In an attempt to develop rapid PCR method the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. The intergenic spacers were amplified by primers complementary to conserved region of 16S and 23S rRNA genes. The intergenic spacer region between the 16S and 23S rRNA genes of V. ichthoenteri were investigated by PCR fragment length typing and DNA sequencing. Analysis of the ISR sequences showed that V. ichthyoenteri contains one types of polymorphic ISRs. The size of ISRs ranged 348bp length and not contains tRNA genes. Mutiple alignment of representative sequences from different Vibrio species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of Vibrio ichthyoenteri. PCR. The specific of the primer was examined using genomic DNA prepared from 19 different Vibrio species, isolated 18group Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

• Fisheries and Aquatic Sciences
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• v.16 no.2
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• pp.125-129
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• 2013

Vibrio ichthyoenteri is a facultatively anaerobic gram-negative bacterium with straight or slightly curved rod morphology. The bacterium is an etiological agent of bacterial enteritis of Olive flounder Paralichthys olivaceus. Only a handful of studies, using a limited number of isolates, have investigated the phenotypic and genetic characteristics of V. ichthyoenteri. We isolated 40 V. ichthyoenteri strains, identified based on the 16S rRNA gene sequence, from the diseased flounder larvae and investigated the API 20E and ZYM profiles. The isolates exhibited highly divergent phenotypic characteristics regardless of sampling time point and location, and fish age. Essential enzymes produced by V. ichthyoenteri seemed to be alkaline phosphatase, leucine arylamidase, and N-acetyl-${\beta}$-glucosaminidase. This study reveals a much greater enzymatic and biochemical phenotype diversity than has been evident to date. These results suggest that a given population of V. ichthyoenteri could be heterogeneous in terms of its phenotypic characteristics.

• Journal of fish pathology
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• v.24 no.2
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• pp.95-102
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• 2011

Normalized resistance interpretation (NRI) analysis for tetracycline was applied to generate information on the epidemiological cut-off value for Vibrio ichthyoenteri isolated from diseased olive flounder (Paralichthys olivaceus) larvae. Thus, 42 strains of V. ichthyoenteri were used to determine minimum inhibitory concentration (MIC) values of tetracycline using Etest. Also, 11 tetracycline resistance related genes were investigated by PCR method. Most tetracycline-resistant strains harbored both tetB and tetM with a few exceptions. NRI-derived mean and 2 SD above the mean of theoretical normal distributions of susceptible isolates were 0.33 mg/L and 1.66 mg/L, respectively. The epidemiological cut-off value for V. ichthyoenteri from the calculations could be set to S ${\leq}$ 2 mg/L. Of the 42 strains, 15 were classified as non-wild type (NWT), and MIC values of the NWT strains vary regardless of tetB and tetM detection, suggesting that there may be other mechanisms involved in tetracycline resistance in this Vibrio species.

• Journal of fish pathology
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• v.19 no.2
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• pp.127-139
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• 2006

From 2002 to 2004, various vibrios were isolated from the olive flounder, Paralichthys olivaceus of culturing size with disease signs. During this survey, it was known that the high proportion of Vibrio ichthyoenteri was occupied among the isolated vibrios. Generally, V. ichthyoenteri is well known as the pathogen of bacterial enteritis of olive flounder larvae. The aim of the present study was the compare the characteristics of two groups of V. ichthyoenteri, culturing sized olive flounder, and larvae of olive flounder showing the intestinal necrosis. The research was focused on the physiology, biochemistry, genetics in the two bacterial groups. The physiological and biochemical characteristics of the tested strains were very similar. The intergenic spacer (IGS) region between the 16S and 23S rRNA genes of 21 isolated strains and 3 reference strains, V. ichthyoenteri, were investigated by PCR fragment length typing and DNA sequencing. After the isolated strains were identified as V. ichthyoenteri, not only phenotypic characteristics of the isolated and reference strains but also homology of 16S-23S IGS of all isolated strains and reference strains as 99.1~100%. The V. ichthyoenteri showed 4 specific 16S-23S patterns and contained no-tRNA, tRNAGlu(TTC) , tRNAIle(GAT) tRNAAla(TGC) type .

• Journal of fish pathology
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• v.23 no.1
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• pp.37-45
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• 2010

In this study, we carried out research on the level of single and multi-drug resistance of bacteria isolated from cultured flounder Paralichthys olivaceus. One hundred sixty one bacteria were isolated from cultured olive flounder Paralichthys olivaceus in Korea and the isolates consisted of Edwardsiella tarda (n=32), Vibrio ichthyoenteri (n=37), Vibrio spp. (n=54), Streptococcus parauberis (n=28) and Streptococcus spp. (n=10). These E. tarda isolates were highly resistant in the order of tetracycline (84.4%) and oxolinic acid (71.9%). V. ichthyoenteri and Vibrio spp. showed resistance ampicillin (94.6% and 81.5%) and tetracycline (56.8% and 42.6%). S. parauberis isolates were resistant ampicillin (57.1%), tetracycline (57.1%) and erythromycin (35.7%). Of the isolates, 84.4% of E. tarda, 73.0% of V. ichthyoenteri, 57.4% of Vibrio spp., 42.8% of S. parauberis and 70.0% of Streptococcus spp. isolates exhibited multi-drug resistance against more than two antibiotics.

• 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.

• Journal of Veterinary Clinics
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• v.34 no.4
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• pp.261-267
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• 2017

The aim of this study was to investigate the antibacterial properties of chitosan silver nanocomposites (CAgNCs) using pathogenic Vibrio ichthyoenteri as a bacterial model. Results of agar disc diffusion and turbidimetric assays showed that CAgNCs could inhibit the growth of V. ichthyoenteri in concentration dependent manner. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CAgNCs were 75 and $125{\mu}g/mL$, respectively. Furthermore, CAgNCs treatment induced the reactive oxygen species (ROS) level in V. ichthyoenteri cells in concentration and time dependent manner, suggesting that it generates oxidative stress, leading to bacterial cell death. The field emission scanning electron microscope (FE-SEM) images of CAgNCs treated V. ichthyoenteri exhibited strong cell membrane damage than un-treated control bacteria. MTT assay results showed the highest cell viability (22%) at $75{\mu}g/mL$ of CAgNCs treated bacteria samples. The results from this study suggest that CAgNCs is a potential antibacterial agent to control fish pathogenic bacteria.

• Journal of fish pathology
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• v.20 no.3
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• pp.281-289
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• 2007

We have investigated lysozyme activity and protectivity of starry flounder, Platichthys stellatus against olive flounder pathogenic bacteria, Edwardsiella tarda, Vibrio ichthyoenteri and Streptococcus iniae in varying salinities, water temperatures and stocking density. Starry flounders were susceptible to E. tarda but not V. ichthyoenteri and S. iniae. Under low salinity condition, the lysozyme activity was decreased a little compared to the control but not significant. The physiological and immune activities were normal up to 26 ℃ culture temperature and 100% stocking density; they were compromised from 29℃ and 200%, respectively.

• Journal of fish pathology
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• v.24 no.2
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• pp.113-120
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• 2011

In this study, we examined antimicrobial activity of the bee venom isolated from honeybee (Apis mellifera L.) against fish pathogenic bacteria, Edwardsiella tarda, Vibrio ichthyoenteri and Streptococcus iniae of cultured olive flounder, Paralichthys olvaceus. The bee venom exhibited significant antimicrobial activity against the both Gram-negative bacteria, E. tarda and V. ichthyoenteri and Gram-positive bacteria, S. iniae. Minimum inhibitory concentration (MIC) and minimun bactericidal concentration (MBC) of the bee venom were 17.6 ${\mu}g$/ml 34.6 ${\mu}g$/ml against E. tarda., and 1.76 ${\mu}g$/ml, 6.8 ${\mu}g$/ml against V. ichthyoenteri, respectively. MIC and MBC of the bee venom were 3.49 ${\mu}g$/ml, 11 ${\mu}g$/ml, respectively against S. iniae. The postantibiotic effect (PAE) of the bee venom was 5 hr, 6 hr, and 7 hr against E. tarda, V ichthyoenteri, and S. iniae, respectively. In addition, its antimicrobial activity was stable under various pH conditions. According to these results, the bee venom showed the excellent antimicrobial activity against the tested pathogenic bacteria.