• Korean Journal of Microbiology
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• v.35 no.3
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• pp.248-252
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• 1999

The PI-oduction of virulence factors such as cholera toxin, heinolysin and hemagglutinin in V cliolerae non-01 and non-0139 were examined. Among 65 strains isolated from environmental and clinical blood sources, 29 (14.6%) strains produced hemolysin only, 35(53.9%) sh.ains produced both hemolysin and hemagglutinin. From one 037 slrain isolated from environmenl, cholera toxin, ctx gene, hemolysin, and hemagglutinin were detected. All of the strains isolated from clinical and environmental sources showed hemolytic activity against human 0 group e~ythrocytes. In inhibition patterns of heinagglotination, 5 of 18 clinical strains (27.8%) were inhibited by less than 1% mannose and galactose, while, among the 47 environmental isolates. hose paltems by less than 1% mannose and galactose 55.4% wel-e inhibited. Thel-ehre, exohamagglutinin positive rate was high in clinical blood isolates but in environnlental sources, the rate was almost similar lo ihe rate or endohemagglutinin positive. These results indicaled that V cholerae non-01 and non-0139 produced various virnlence factors such as cholera toxin, hemolysin, and hemagglutinin but not a single factor. Further studies are need for epidemiological or bacteriological shtdies of V cholerae 037 isolated from environment.

• KSBB Journal
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• v.23 no.2
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• pp.125-130
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• 2008

We investigated the biological activities of ethanol extract from the seawater algae, Chlorella elliposidea C020 such as antibacterial activity, anti-oxidant activity, tyrosinase inhibitory activity and hemolytic activity against human erythrocytes. Extract was obtained from various solvent, methanol, ethanol, acetone and ethanol + acetone (1:1, v/v%), 95% ethanol proved to be best extraction solvents. The contents of ethanol extract were higher in freeze-dried sample than that in frozen-thawing. Antibacterial activities of ethanol extract showed strong inhibitory effect against Bacillus subtilis PM125, Bacillus licheniformis and fish pathogenic bacteria, Vibrio parahaemolyticus KCTC2471 and Edward tarda NUF251. However, this extract didn't worked against antifungal activity against Candida albicans KCTC1940. And, ethanol extract was without hemolytic activity against human erythorocytes. The ethanol extract showed 75% of free radical scavanging effect on 2.0 mg/mL using DPPH method. In tyrosinase inhibition assay of ethanol extract, $IC_{50}$ (Inhibition Concentration) was measured as 10.87 mg/mL. Conclusionally, ethanol extract of Chlorella elliposidea C020 has good candidate for bioactive materials.

• The Journal of the Korean Society for Microbiology
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• v.35 no.3
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• pp.251-261
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• 2000

V. vulnificus is an estuarine bacterium which causes septicemia and shock in susceptible patients. The organism produces a hemolytic cytolysin (VvH), which has a membrane damaging effect on erythrocytes. To clarify the mechanisms by which VvH might contribute to virulence, we examined its effect on macrophages. When mouse peritoneal macrophages were harvested and co-cultured with hemolysin-positive V. vulnificus strains (100 bacteria/cell), about 60% of the macrophages were killed; macrophages were not killed when co-cultured V. vulnificus strain CVD 707, a VvH-negative deletion mutant. Exposure of macrophages to filtered culture supernatants (2.5 HU/ml) and purified VvH (3 HU/ml) resulted in an increase in dead cells (80 and 90%, respectively), as determined by the trypan blue dye exclusion method and LDH release from macrophages was also increased (70 and 65.5%, respectively). The cytotoxic effect of VvH on macrophages was both the dose- and time-dependent. The VvH caused damage to the macrophage membrane and was blocked significantly by preincubation with cholesterol (p<0.01). Fetal bovine serum showed remarkable inhibition of VvH synthesis by V. vulnificus and inhibited VvH activity in culture supernatant. Cell viability was increased by 35% (p<0.01) and LDH release decreased by 28% (p<0.01) when macrophages were incubated with V. vulnificus (100 bacterial cell) in DMEM-10% FBS for 2 hr. Bacterial clearance activity of mice against V. vulnificus CVD 707 was decreased by pretreatment with 10 HU of VvH. This result suggests that the VvH can impair the membrane of macrophages and may playa role in the pathogenesis of V. vulnificus septicemia.

• The Journal of the Korean Society for Microbiology
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• v.18 no.1
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• pp.1-9
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• 1983

The author studied on the distribution of V. parahaemolyticus among sea water, sea mud, and various marine products in Jeju, Keoje, Namhae, Yockji, Busan, and Masan on the southern seas of Korea from winter in 1981 to summer in 1982. The author studies for the isolated strains to bacteriological identification Kanagawa phenomenon(hemolytic activity) on Modified Wagatuma blood agar plates and serotypes with anti V. parahaemolyticus. The results obtained were as follows:. 1. V. parahaemolyticus was isolated in 713 strains(28.3%) among 2519 total specimens of sea water, sea mud, and various marine products. 2. The isolation rates of V. parahaemolyticus in summer season were higher than in spring and winter season. Above results were 304 strains(32.6%) among 932 specimens in summer, 160 strains (28.1%) among 570 specimens in spring, 149 strains(14.6%) among 1017 specimens in winter, respectively. 3. The hemolysis on Modified Wagatuma agar added human erythrocytes was 66.0% of positive Kanagawa phenomenon, and was 34.0% of negative Kanagawa phenomenon, respectively. 4. The distributions of serotypes of V. parahaemolyticus isolated were from KI to K VIII of K pooling antisera. The results were 6 strains(6.6%) on K 1 of K I, 14 strains(l5.5%) on K 17 of K II, 26 strains(28.8%) on K 28 of K IV. 10 strains(11.1%) on K 32 of KV, 4 strains(4.4%) on K 39 of KV, 8 strains(8.8%) on K 42 of K VI, 2 strains(2.2%) on K 48 of K VII, 1 strain(1.1%) on K 50 of K II and 7 strains(7.7%) on K 55 of K VII, respectively. 5. V. parahaemolyticus was more frequently isolated from sea mud than sea water and various marine prdoucts in winter season. 6. There was no great difference as far as the distribution of V. parahaemolyticus concerned in Jeju, Keoje, Namhae, Yockji, Busan and Masan of the southern seas of Korea.

• Journal of Life Science
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• v.22 no.5
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• pp.642-649
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• 2012

The southern coast of Korea is important for the ark shell ($Scapharca$ $broughtonii$) aquaculture, but the productivity was rapidly reduced during the previous decade by mass mortality. To overcome this economic loss, investigations only focused on environmental factors, and microbiological researches were performed insufficiently. In this study, two sites (Gangjin and Jinhae bay) were selected for their high and low rate of mortality, respectively, and the existence of microflora from underwater sediments in the bodies of $S.$ $broughtonii$ was analyzed. We screened the whole body of each sample and chose unique colonies, which exhibit alpha- and beta-hemolytic activity, for identification. The microflora in $S.$ $broughtonii$ was less variable than sediments, and restricted species were isolated. We identified 17 genera of 88 species and 16 genera of 64 species from the two bays, respectively. A major proportion was comprised of $Bacillus$ species, with the $Bacillus$ $cereus$ group being the most common species among the $Bacillus$ strains, while $Paenibacillus$, $Lynsilbacillus$, and $Vibrio$ species were the second most abundant species. At the genus level, there were no significant microbial differences between the two coastal regions. 64 species were isolated from rare site (Jinhae bay), but more species (88) with greater variety were isolated from the frequent site (Gangjin bay). Therefore, it was assumed that the cause of mass mortality lay in the difference in specie-level diversity, and conducting investigations on the diagnosis of pathogenic species by challenging tests using isolated unique species.

• Journal of Food Hygiene and Safety
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• v.21 no.4
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• pp.213-217
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• 2006

The halophilic bacterium Vibrio vulnificus is known to be a foodborne pathogen that causes septicemia in human. V. vulnificus infection is characterized by the high fatality rates and the primary attack against a person who have underlying diseases such as liver cirrhosis. However, there is no effective treatment for V. vulnificus septicemia except for classical treatments such as antibiotics. Recently, it has been known that lipoprotein (LDL) plays a major role in the protection against infection and inflammation. Consequently in this paper we analyzed the effects of LDL on V. vulnificus septicemia. We purified V. vulnificus cytolysin, a major virulent factor of V. vulnificus infection and measured inhibitory effects of mouse serum, cholesterol, and LDL on its hemolytic activity. Next experiments were performed to investigate whether LDL has a protective role against septicemia induced by V. vulnificus in mice. Intraperitoneal injection of LDL (1mg as protein) into mice 3hr before V. vulnificus $(1\times10^6\;CFU)$ injection, and V. vulnificus -induced lethality was determined. For the determination the relationship between LDL or cholesterol and prognosis, we determined serum levels of cholesterol and lipoprotein from V. vulnificus septicemia patients (n=15) who had visited the Chonbuk National University Hospital in Chonju. V. vulnificus cytolysin -induced hemolysis of mice erythrocytes was completely inhibited by serum, cholesterol, and low-density lipoprotein. V. vulnificus- induced lethality of mice injected with LDL showed only 40% compared to 100% of control. In survival groups (n=4) of V. vulnificus septicemia patients (n=15), their serum LDL and cholesterol revealed normal levels ($153.3{\pm}40.7,\;LDL;\;190.8{\pm}16.3$, Total cholesterol). However, in death groups (n=11) showed very low levels ($35.6{\pm}13.9,\;LDL;\;59.2{\pm}15.1$, Total cholesterol). Our study indicates that cholesterol and LDL are a prognosis indicator of V. vulnificus septicemia as well as an inhibitor of virulent action of V. vulnificus cytolysin. We suggested that the serum levels of cholesterol or LDL would be major index in the treatment and prevention of V. vulnificus septicemia.

• Journal of Life Science
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• v.28 no.11
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• pp.1301-1315
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• 2018

We purified an antimicrobial peptide from the acidified hemocyte extract of Mytilus coruscus by $C_{18}$ reversed-phase high-performance liquid chromatography (RP-HPLC). The peptide was 4041.866 Da based on matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS) and the 25 amino acids of the N-terminus sequence were identified. Comparison of this sequence of the purified peptide with the N-terminus sequences of other antimicrobial peptides revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We also identified a 312 bp open-reading frame (ORF) encoding 103 amino acids based on the obtained amino acid residues. The nucleotide sequence of this ORF and the amino acid sequence also revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We synthesized two antimicrobial peptides with an alanine residue in the C-terminus, and designated them mytilin B1 and B2. These two antimicrobial peptides showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus and Streptococcus parauberis (minimal effective concentration, MECs $41.6-89.7{\mu}g/ml$), gram-negative bacteria, including Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Providencia stuartii, Pseudomonas aeruginosa, and Vibrio ichthyoenteri (MECs $7.4-39.5{\mu}g/ml$), and the fungus Candida albicans (MECs $26.0-31.8{\mu}g/ml$). This antimicrobial activity was stable under heat and salt conditions. Furthermore, the peptides did not exhibit significant hemolytic activity or cytotoxic effects. These results suggest that mytilin B could be applied as alternative antibiotic agent, and they add to the understanding of the innate immunity of hard-shelled mussels.