• Korean Journal of Veterinary Research
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• v.32 no.2
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• pp.181-194
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• 1992

Two hundred and eighty nine strains of Yersinia enterocolitica isolated from healthy pigs were tested for the presence of 40~50 Megadalton virulence-associated plasmids and plasmidmediated in vitro virulence-associated properties, i.e., congo red uptake, calcium dependency, autoagglutination, CRMOX reaction, crystal violet binding and pyrazinamidase reaction. The correlationships between in vitro virulence-associated properties and the presence of 220 Kdalton outer membrane protein(OMP) were examined in strains with or without virulence-associated plasmids. The correlationships between the presence of plasmids on the production of the OMP and the expression of in vitro virulence-associated properties were studied with $CRMOX^+$ strains and acridine orangecured $CRMOX^-$ mutants. The results were as follows : 1. Of the in vitro virulence-associated tests with 289 strains of Y enterocolitica, 275 strains (95.2%) were positive for pyrazinamidase test, and followed by in order of crystal violet binding test, 226 (79.2% ) ; CRMOX test, 190 (65.7%) ; autoagglutination test, 1.85(64.0%) : calcium dependency test, 86 (29.8%) and congo red uptake test, 47(16.3%). 2. The correlationship between autoagglutination and CRMOX test(r=0.90) was highly significant (p<0.01). 3. In 190 strains(65.7%) bearing the virulence-associated plasmids(MW 40~50 Mdalton), the correlation between the presence of plasmids and their in vitro virulence-associated properties were highest with CRMOX test(r=0.93) and followed by in orders of AAG test(0.81), CV test(0.46), PYZ test(0.37) and CD test(0.18), but no correlationship between the presence of plasmids and CR test(-0.11). 4. The $CRMOX^+$ strains produced the 220 Kdalton OMP when they were cultured at $37^{\circ}C$, but not at $26^{\circ}C$. The presence of 220 Kdalton OMP was correlated significantly with in vitro virulence properties and the presence of virulence-associated plasmid, respectively. 5. In the isogenic $CRMOX^-$ mutant strains, of which plasmid were cured by treatment with acridine orange not only in vitro virulence-associated properties(CR 100%, CD 100%, AAG 82.6%, CV 58.3%) disappeared but also 220 Kdalton OMP(100%) was not produced. These results indicate that the positive CRMOX reaction is plasmid-mediated and the CRMOX test is potential as an in vitro virulence tests with Y enterocolitica.

• Korean Journal of Microbiology
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• v.26 no.3
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• pp.262-269
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• 1988

In order to investigate the virulence of Y. enterocolitica isolated in Korea, all necessary experiments were done including several virulence determinating tests-autoagglutination test, calcium-dependency test, HeLa cell invasion test, Sereny test, crystal violet binding test, and electrophoresis for plasmid pattern. The obtained results are as follows: The virulent strains of Y. enterocolitica revealed positive reactions on autoagglutination test, cacium-dependency test, and drystal violet binding test, while the avirulent strains did not. A positive reaction was observed only at $37^{\circ}C$ implying that the expression of virulence is temperature-dependent. In Sereny test, the standard reference virulent strain (serotype 0:8) showed positive reactions while the virylent experimental strains (serotype 1:3, 0:9) revealed negative results, which indicates that the virulence of Y. enterocolitica in experimental animals varied according to their serotypes. Most of the virulent strains contained 36-38Mdal plasmids, but the avirulent strains did not. In addition, it was noted that autoagglutination, calcium-dependency, and crystal violet binding were related to the presence of plasmids.

• Korean Journal of Veterinary Research
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• v.32 no.3
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• pp.369-376
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• 1992

This paper dealt with plasmid DNA profile in 98 Salmonella(S) isolated from pigs and cattle sources in Taegu, Gyeongbook and Gyeongnam during the period from 1984 to 1987. Also we were studied for restriction enzyme analysis of the plasmid DNA, and mouse infection, Sereny test and normal setum resistance test in guinea pig for S typhimurium and S enteritidis harbored or cured 60 megadalton(Md) plasmid and 36 Md plasmid, respectively. Of the 13 Salmonella isolated from cattle, 7 Salmonella harbored one or more plasmids and molecular sizes of the large plasmids were 60 Md for S typhimurium and 36 Md for S enteritidis. Of the 85 Salmonella isolated from pigs, 47 Salmonella were confirmed as being one or more plasmids, and all the S typimurium stains harbored 60 Md plasmid. In enzyme digestion with 8 types of restriction endonuclease for 60 Md plasmid DNA of S typhimurium, cleavage patterns were varied to enzymes, and the DNA was segmented into 4 to 15 fragments. In restriction enzyme analysis of 36 Md plasmid DNA obtained from four strains of S. enteritidis, the DNA showed the same cleavage patterns obtained with Eco RI, Hind III and Bam H I, and was segmented into 3 to 5 fragments. In virulence for mice by measuring the 50% lethal dose ($LD_{50}$), the $LD_{50}$ values obtained for 60 Md virulence-associated plasmid harbored strains of S typhimurium and 36 Md virulence-associated plasmid of S enteritidis were up to $10^4$-fold lower than the values obtained for the plasmid-cured strains of the same serotype. Only the plasmid harbored strains were resistant to the bactericidal activity of 90% guinea pig serum, and only they gave positive responses in sereny test. We suggested that their plasmid DNA might be associated with virulence for mice.

• Korean Journal of Veterinary Research
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• v.51 no.1
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• pp.15-20
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• 2011

In this study, we isolated 12 of Brucella (B.) spp. from cattle, which have been positive in Rose Bangal test and tube agglutination test in Gyeongbuk province in 2009. According to AMOS PCR analysis, isolated 12 strains were identified as B. abortus. Murine derived macrophage, RAW 264.7 cells, were infected with isolated 12 strains or reference strain (B. abortus 544), and bacterial internalization were characterized. According to these results, we divided the isolated strains into the following three groups: class I, lower internalization than that of B. abortus 544; class II, similar internalization to that of that of B. abortus 544; class III, higher internalization than that of B. abortus 544 within RAW 264.7 cells. Furthermore, intracellular growth, bacterial adherent assay, LAMP-1 colocalization, virulence in mice and surface protein pattern were characterized. From these results, representative strains of class III showed lower LAMP-1 colocalization, higher adherent efficiency, higher virulence in mice than those of B. abortus 544, and showed different pattern of surface proteins. These results suggest that B. abortus field strains, isolated from cattle in Korea, possess various virulence properties and higher internalization ability of field strain may have an important role for its virulence expression.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.515-519
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• 2004

Listeria spp. were isolated from a total of 402 naturally contaminated domestic ready-to-eat (RTE) vegetable samples by the conventional Food and Drug Administration protocol and confinned by API-Listeria kit. Also, the susceptibility to 12 antibiotics, polymerase chain reaction (PCR) assay for virulence gene of pathogenic Listeria monocytogenes isolates, and in vitro virulence assay using myeloma and hybridoma cells from murine and human sources were tested. Among the samples, 17 samples (4.2％) were found to be contaminated with Listeria species. Among the 17 strains of Listeria spp. isolates, only 2 strains (11.8％) of L. monocytogenes and 15 strains (88.2％) of L. innocua were identified. Antibiotic susceptibility test showed that the Listeria spp. isolates were very susceptible to the antibiotics tested, except for nalidixic acid. Among 17 strains of Listeria spp., PCR analysis showed that 2 strains of L. monocytogenes isolates proved to have a virulence hly gene, but none of L. innocua had the hly gene. Also, hybridoma Ped-2E9 cells assay showed that only L. monocytogenes isolates killed approximately 95-99％ hybridoma cells after 6 h, but L. innocua isolates had about 0-5％ lethal effect. These results indicate that PCR assay with hly primer or hybridoma Ped-2E9 cells assay could be used as a good monitoring tool or in vitro virulence test for L. monocytogenes.

• Journal of environmental and Sanitary engineering
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• v.15 no.4
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• pp.105-113
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• 2000

We have isolated 65 strains(2.0%) of Y. enterocolitica among 3,219 water samples from 380 spring water sites in Seoul from 1994 to 1999. The biochemical characteristics of isolated strains revealed that TSI was A/A, urea, M.R.($37^{\circ}C$), nitrate, motility($37^{\circ}C$), sorbitol, maltose, manitol, arabinose, mannose, trehalose, xylose were positive(100%) and H$_2$S, arginine, lysine, oxidase, citrate, V.P.($37^{\circ}C$), DNase, motility($37^{\circ}C$), dulcitol, adonitol, lactose and raffinose were negative(100%). In in vitro virulence test, positive rate of AAG and CRMOX were 9.2% and 4.6%, respectively. However in the virulence gene detectable gene detectable test by multiplex-PCR using ail, yst, virF genes, 65 strains were all negative, meaning that Y.enterocolitica strains from domestic spring water were not detected for the virulence. Otherwise, mutiplex-PCR which using ail, yst and subgenus-specific primer pair was the best for identifying the virulence of Y. enterocolitica.

• Korean Journal of Veterinary Research
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• v.29 no.2
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• pp.69-73
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• 1989

Nine strains of Yersinia species isolated from pigs and dogs in Korea, comprising 5 strains of Yersinia enterocolitica, 2 strains of Y kristensenii and each strain of Y pseudotuberculosis and Y intermedia, were examined for the presence of virulence-associated plasmids, calcium dependency and provocation of guinea pig conjuntivitis($Ser{\acute{e}}ny$ test). Three strains of Y enterocolitica isolated from pigs were positive in calcium dependency and harbored one plasmid of about 45 megadalton, but negative in $Ser{\acute{e}}ny$ test.

• Research in Plant Disease
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• v.21 no.2
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• pp.103-106
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• 2015

Isolates of the rice blast fungus show a range of tissue-specificities infecting leaves, nodes, neck and panicles. Although neck and panicle blast cause significantly greater yield losses than the leaf blast, virulence tests of the blast isolates have been performed only rice leaves instead of neck and panicles. In this study, we have developed a virulence test method for neck and panicle blast. We selected three representative isolates from each of leaf, neck, and panicle blast. We observed that severe disease lesions developed on the neck and the panicles when the infected rice plants were incubated in a dew chamber for 48 h instead of 24 h when tested on leaves. Unlike the leaf blast, a typical lesion on the neck and panicles appeared after 14 days post-infection as opposed to 7 days with leaf blast. This method will be applied to examine tissue-specificity of the rice blast fungus isolates.

• The Journal of the Korean Society for Microbiology
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• v.35 no.4
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• pp.317-324
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• 2000

Candida albicans is one of the most frequently isolated fungal pathogens in human. Recently, the prevalence of candida infection has markedly increased, partially due to the increase of immunocompromised hosts. Proposed virulence factors of the pathogenic Candida are the ability to form hyphae to adhere to epithelial cell surfaces, and to secrete acid proteinases and phospholipases. We measured the relative cell surface hydrophobicity (CSH) and the ability of proteinase production (PROT), phospholipase production (PLase), adherence to host epithelium (ADH), and hyphal transition (Germ). The relative risk of virulence factors was analyzed by lethality test in murine model of hematogeneously disseminated candidal infection. According to Cox's proportional hazard analysis, the statistically significant virulence factors were PROT, ADH, and CSH. PROT was the highest risk factor of them. To evaluate the applicability for the diagnosis and treatment of Candidiasis, we examined the protective effect of the active and passive immunizations with the materials purified from virulence factors and antibodies to them in Candia-infected mice model. The mean survival times of active and passive immunized groups were slightly longer than those of non-immunized groups.

• Korean journal of applied entomology
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• v.13 no.1 s.18
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• pp.32-32
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• 1974