• Journal of fish pathology
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• v.26 no.3
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• pp.255-263
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• 2013

The innate immune response is fundamental defense response of vertebrates and invertebrates. Especially, the innate immune response important for larvae that lack of resistance to infectious diseases in the early stages. Galectin is one of the kinds of lectin and presents in the fish mucous that involves innate immune response. Galectin have been studied from various fishing species, but expression analysis of galectin is still unclear during early developmental stage in olive flounder. In this study, we investigated gene expression of galectin-1 from various developmental stage and tissues. We excised several tissues including the muscle, fin, eye, gill, brain, stomach, intestine, kidney, spleen and liver from adult olive flounder and confirmed gene expression of galectin-1 using RT-PCR and quantitative real-time PCR. Expression of galectin-1 was significantly higher in muscle, stomach and intestinal tissue than other tissue in adult fish (5 and 29 months). Also, galectin-1 gene was detected from 0 DAH and gradually increased to 35 DAH and since then decreased after stomach development period. Induction of galectin-1 during the early developmental stage suggest that muscle, fin and eye tissue is formed and begins the secretion of galectin this period. In addition, increased expression levels at 35 DAH suggest that due to complete formation of stomach and intestine, increase of secretion and activation of enzyme. This study shows that expression of galectin-1 during early developmental stages and adult period in olive flounder and can be expect that galectin-1 play essental role in the innate immune system throughout the whole life time. Galectin-1 is primary barrier such as skin and digestive tissue against pathogen infection, also digestive tract developmental period is important for pathogen invasion can be expected that it will serve. Mass mortality due to the disease in seed production is continuing damage, therefore these result will be meaningful about infectious disease during early developmental stages as a basic data for the study.

• Annals of Clinical Microbiology
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• v.18 no.2
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• pp.37-43
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• 2015

Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.526-529
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• 2012

The present study was aimed to investigate the effect of a probiotic, Enterococcus faecium, on the immune responses against infection with the marine fish pathogen Lactococcus garvieae in olive flounder (Paralichthys olivaceus). The immune responses were assessed by lysozyme activity, complement activity, protease activity, and expression of proinflammatory cytokines by RT-PCR. The lysozyme and complement activities were increased between 9 to 15 and 9 to 13 days, respectively, and antiprotease activity was slightly elevated after 5 days of probiotic treatment. The TNF-${\alpha}$ and IL-$1{\beta}$ expressions were observed from kidney and spleen. The results of this study reveal that E. faecium induces immune-responsible materials and protects olive flounder from lactococcosis.

• Biomolecules & Therapeutics
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• v.6 no.2
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• pp.191-198
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• 1998

Vsrio vulnificus is an estuarine gram-negative human pathogen that affects people with chronic hepatitis, alcoholic cirrhosis, diabetes mellitus or other underlying diseases. V. vulnificus infection is mediated primarily by consumption of raw fish or by exposure of pre-existing wounds to seawater, causing permanent tissue damages or fatal septic shock. We have been developing a vaccine against V. vulnificus composed of whole cell Iysate of a V. vulnificus O-antigen serotype 4 strain. Oral administration of the V. vulnificus;oral vaccine;immunogenicity;protective efficacy vaccine elicited a high serum antibody response in rabbits. The induced antibodies were reactive not only to the homologous strain but also to heterologous O-antigen serotype strains, indicating cross-reactivities among serotypes. Western blot analysis revealed that the antibodies are mainly specific for outer membrane proteins (OMPs) and reacted equally well with OMPs purified from 9 O-antigen serotypes. The rabbit antisera showed opsonophagocytic killing activity against heterologous strains as well as the homologous strain. Passively transferred rabbit antisera into mice were protective against a lethal V. vulnificus infection. These data demonstrate that oral administration of the V. vulnificus vaccine induced a systemic antibody response which had a protective efficacy against V. vulnificus infections, suggesting that this vaccine preparation could be used to develop an oral vaccine against V. vulnificus.

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.535-539
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• 2016

The prevalence of pathogenic bacteria such as Streptococcus parauberis (Sp), Streptococcus iniae (Si), and Edwardsiella tarda (Et) in flounder fish farms in Jeju Island and their management by gallium treatment was studied. Sp, Si, and Et were found to exhibit a low rate of cell growth and high biofilm formation. Hence, in the present study, cell growth and biofilm formation were measured spectrophotometrically 72 h after the addition of different concentrations of gallium (2, 4, or 8 mg/ml). In addition, cell death was measured by resazurin and propidium iodide staining assays. The results showed that bacterial cell death increased and biofilm formation decreased with an increasing concentration of gallium. Hence, the present study signifies that the use of gallium against bacterial pathogens could be useful for disease management in flounder farms.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.4
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• pp.365-369
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• 2001

Streptococcus spp. of bacterial pathogen of fish were isolated from the cultured flounder (Paralichthys olivaceus) in fish farm of Jeju Island. Clinical signs of the infected flounder which are the most commons symptoms are as follows: erratic swimming, darkening of the body colour, unilateral or bilateral exophthalmia, corneal opacity, hemorrhages in the opercular and the bases of the fins, and the ulceration of the body surface. Biochemical characteristics of pathogenic fish Streptococcus spp, were gram positive, spherical form, catalase negative, oxidase negative and $\beta$-haemolytic, respectively, The viable cells counted from the tissue of the diseased flounder were the largest in the order of the ulcer, the kidney, the blood and the brain, The drugs used were ampicillin, ciprofloxacin, doxycycline, gentamycin, tetracycline, erythromycin, streptomycin and oxytetracycline, Streptococcus spp. were found to be sensitive to ampicillin, ciprofloxacin, doxycycline and gentamycin, but were resistant to tetracycline, erythromycin, streptomycin and oxytetracycline. The pathogenicity of Streptococcus spp. on the cultured flounder with an abdominal cavity injection was high. The haemolytic activity of the toxin against the sheep red blood cells reached the maximum after 30 min incubation at $37^{\circ}C$ or $50^{\circ}C$. The toxin showed highest activity at pH $5.5\sim6.5$.

• Korean Journal of Food Science and Technology
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• v.31 no.4
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• pp.1035-1043
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• 1999

Bacteriocins are natural antimicrobial compounds produced by many microorganisms associated with foods, so that there is currently much interest in their use as food biopreservatives. Goal of this study was to partially evaluate lacticin NK24 as a food biopreservative by showing antimicrobial activity of L. lactis NK24 and lacticin NK24 against food-borne spoilage and pathogenic bacteria, respectively. Lactic acid bacteria NK24 isolated from jeot-gal, Korean fermented fish foods, was tentatively identified as Lactococcus lactis and showed broad spectrum of activity against all of spoilage and pathogenic bacteria tested by deferred method. Bacteriocin production in jar fermenter was detected at the mid-log growth phase, and reached the maximum at the early stationary phase, but decreased after the stationary phase. Lacticin NK24 was partially purified by 75% ammonium sulfate precipitation followed by subsequent dialysis. This partially purified lacticin NK24 showed antimicrobial activity against Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Clostridium perfringens, some bacilli, Listeria monocytogenes, Listeria ivanovii, Sphin-gomonas pausimobilis, Escherichia coli and Pseudomonas aeruginosa. Thus, lacticin NK24 examined in this study show promise as a biopreservative be-cause of their broad spectrum of activity.

• Korean Journal of Ichthyology
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• v.29 no.1
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• pp.62-68
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• 2017

The Alaska pollock (Gadus chalcogrammus) belongs to the family Gadidae; it is a cold water fish, and has been developed as a novel aquaculture species in Korea. In this study, we describe ongoing surveillance for aquatic animal pathogens based on growth stages. We investigated bacterial flora in rearing water, and monitored pathogens; we also analyzed histopathological traits of abnormal fish. In rearing water, the total bacterial counts were $2.1{\times}10^3cfu/mL$ and Vibrio spp. (52%) were predominant in the larvae stage. In the juvenile and adult stages, the total bacterial counts were $3.4{\times}10^3$ and $3.2{\times}10^2cfu/mL$, respectively (with Pseudomonas sp. as the predominant species; 90% and 52%). This result revealed that the bacterial flora in rearing water changed depending on the feeding types. No virulent-bacteria or problematic viruses (VHSV, viral hemorrhagic septicemia virus; NNV, nervous necrosis virus; MBV, marine birnavirus) were detected from outwardly healthy fish using either culture or PCR assay. Some juveniles (less than 5%) had gas bubbles on the gill lamellae, degeneration of the corneal epithelium, and choroid gland degeneration, suggesting that these symptoms were caused by external injury and secondary infection by opportunistic bacteria. Disease management is important to cope with disease emergence in the novel aquaculture species Alaska pollock.

• Journal of Fisheries and Marine Sciences Education
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• v.28 no.6
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• pp.1715-1723
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• 2016

The heat extracts of six kinds of medicinal herbs (Scutellaria baicalensis, Sophora flavescens, Citrus unshiu pericarpium, Lonicera japonica, Perilla frutescens, Benincasa hispida) were tested for non-specific immune response and disease resistance effects related with fish diseases on olive flounder, Paralichthys olivaceus. The preventive effects of 6 herbal mixtures against bacterial disease on cultured flounder were examined follow as feeding EP absorbed with the heat extract of six kinds of medicinal herbs. For feeding trial for 12th week, weight gain and serum analysis of fish fed various groups were not significant differences. Lysozyme activity of the 0.01 % treated group on 4th week showed significant increase. Histopathology of the administrated group for feeding period showed no particular signs of tissue degeneration. At 0.01% oral experimental group, relative percent survival (RPS) were only 50% to 75% for four weeks and eight weeks group by intraperitoneal injection with E. tarda. The results suggest that heat extracts of six kinds of medicinal herbs (0.01%) would be effective to enhance the nonspecific immunity and protective ability of olive flounder against fish pathogen.

• Journal of Life Science
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• v.12 no.6
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• pp.669-675
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• 2002

Introduction of sliced raw fish meat(SRFM) to fast food business has been considered seriously. However bacteria causing food poisoning should be controlled. Organic acids such as vinegar and lactic acid used in the sauce for SRFM were evaluated for their antibacterial activities. At low concentration levels of vinegar and lactic acid exerted strong antibacterial activities toward Vibriu sp.. In contrast, in case of Salmonella typhimurium and Escherichia coli O157:H7 low anitbacterial activities were observed even at relatively high concentrations. Minimum inhibitory concentrations(MIC) of vinegar for V. vulnificus, V. cholerae non-O1, V. parahaemolyticus, V. mimicus, S. typhimurium and E. coli O157:H7 were 16, 18, 16, 12, 26, and $20{\mu}\ell /m\ell, respertively. MIC of lactic acid for V. vilnificus, V. cholerae non-O1, V. parahaemolyticus, V. mimicus, S. typhimurium and E. coli O157:H7 were 20, 25, 25, 25, 40, and $35{\mu}\ell /m\ell, respectively. In case of vinegar bactericidal concentration upon 10 second contact for V. vulnificus, V. cholerae non-O1, V. parahaenolyticus, V. mimicus and E. coli O157:H7 were 8, 14, 10, 4, and 48%, respectively; however, even at 50% colony of S. typhimurium was observed. In case of lactic acid any colony was observed for V. vulnificus, V. cholerae non-O1, V. parahaemolyticus, V. mimicus, S. typhimurium and E. coli O157:H7 at the concentration of 2, 3, 4, 3, 14, and 17%, respectively. Vinegar and lactic acid of low concentration inhibited the growth of Vibrio sp., food poisoning pathogen in SRFM; in contrast, at high concentration these organic acids inhibited Salmonella sp. and Escherichia sp., food poisoning pathogen in other than SRFM.