• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.6
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• pp.918-926
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• 2021

We isolated 28 Vibrio vulnificus strains from seawater and fisheries and investigated the positive rate of eight virulence genes. Additionally, we evaluated the susceptibility of these strains to 25 antimicrobials. The positive rates of fur, vvhA, tcp, rtxA, vcgC, viuB, vvp, and acfA were 100, 92.9, 92.9, 67.9, 64.3, 25.0, 14.3, and 7.1%, respectively. A disk diffusion susceptibility test revealed that, all the investigated strains had the highest resistance to amoxicillin and oxacillin, followed by that to streptomycin (96.4%), cefoxitin (92.9%), clindamycin (82.1%), amikacin (67.9%), vancomycin (46.4%), nalidixic acid (7.1%), penicillin G (7.1%), and ampicillin (3.6%). Moreover, they were susceptible to 10 other antimicrobials, including cefotaxime, chloramphenicol, erythromycin, gentamicin, and rifampicin. Notably, amoxicillin, oxacillin, and streptomycin had average minimum inhibitory concentrations of 132.6, 603.4, and 23.1 ㎍/mL against V. vulnificus, respectively. These observations provide new insights regarding the necessity for sanitation of commercial fisheries and can potentially, help reduce the risk posed by fisheries contaminated with bacteria resistant to antimicrobials.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.3
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• pp.220-226
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• 2014

Sixty-seven Vibrio parahaemolyticus isolates from surface seawater from the Wando area, on the southern coast of Korea, were analyzed for their susceptibility to 15 different antimicrobials and the presence of virulence genes. According to the disk diffusion susceptibility test, all of the strains studied were resistant to ampicillin and oxacillin, while decreasing percentages were resistant to vancomycin (64.2%), streptomycin (56.7%), amikacin (31.3%), kanamycin (22.3%), cephalothin (20.9%), erythromycin (10.4%), ciprofloxacin (4.5%), and tetracycline (3.0%). All of the strains were susceptible to five antimicrobials: chloramphenicol, gentamycin, nalidixic acid, sulfamethoxazole/trimethoprim, and trimethoprim. Fifty-nine isolates (88.1%) were resistant to three or more classes of antimicrobial and defined as multidrug resistant, and two strains were resistant to seven antimicrobial agents. The minimum inhibitory concentration (MIC) of the 67 V. parahaemolyticus isolates to ampicillin and oxacillin ranged from 512-2,048 and $64-512{\mu}g/mL$, respectively. All 67 isolates were also examined for the presence of the tdh and trh virulence genes using the polymerase chain reaction (PCR). However, no isolates possessed either tdh or trh. The VPA0477 (${\beta}$-lactamase) gene, present in all of the tested strains, was validated as a new specific marker gene in PCR assays for the accurate detection and identification of V. parahaemolyticus.

• Journal of Food Hygiene and Safety
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• v.29 no.4
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• pp.316-321
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• 2014

The aim of study was to investigate the virulence profile of Escherichia coli O157:H7 bacteriophages isolated from sewage and livestock stools. Among 23 E. coli O157:H7 bacteriophages, 14 strains were isolated from sewage and 9 were from animal stools collected from 10 livestock farms in Korea. For each bacteriophage DNA sample, the presence of stx1, stx2, eae, aafII, ial, elt, estI, estII, astA, afa, and cnf was examined by polymerase chain reaction. The detection rate of eae, stx2, estI, astA, and ial was 100%, 69.6%, 13.0%, 13.0%, 8.7%, respectively. While all E. coli O157:H7 bacteriophages isolated from stools carried eae+stx2, stx2+eae, eae+astA, eae, stx2+eae+estI, eae+estI, stx2+eae+ial, and eae+ial were observed in bacteriophages isolated from sewage. As several plasmid-carrying virulence factors (estI, astA, and ial) were found in E. coli O157:H7 bacteriophages obtained from sewage and stools, the microbial safety of bacteriophages should be investigated in further study.

• Journal of Microbiology
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• v.38 no.1
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• pp.1-7
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• 2000

Eight strains of Aeromonas hydrophila isolated from diseased trout in Korea were characterized and compared with an American type strain by various methods including biochemical and physiological tests, PCR, randomly amplified polymorphic DNA (RAPD), plasmid profiling, and gel electrophoresis of total, membrane, and extracellular proteins. Virulence factors such as surface array proteins, cytotoxin, hemolysin, haemagglutinin, and protease were also investigated. The Korean strains showed heterogeneity in Iysine decarboxylase production, utilization of various carbon sources, and production of acetoin. Five strains had the same profiles of total and membrane proteins. Six strains haemagglutinated with trout red blood cells (RBCs) which was inhibited by fucose, galactose, and mannose, except for No. 1 where haemagglutination was inhibited by only galactose and mannose, but not by fucose. Four isolates haemagglutinated with human RBCs which was inhibited by fucose and mannose yet not by galactose. The type strain haemagglutinated only with trout RBCs which was inhibited by fucose, galactose, and mannose. Every isolate secreted protease, hemolysin, cytotoxin, and siderophore, but no enterotoxin. Results showed that the Korean isolates, except for No.7, had very different biochemical and molecular characteristics from those of the American type strain.

• The Plant Pathology Journal
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• v.38 no.4
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• pp.410-416
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• 2022

Erwinia amylovora, the causal agent of fire-blight disease in apple and pear trees, was first isolated in South Korea in 2015. Although numerous studies, including omics analyses, have been conducted on other strains of E. amylovora, studies on South Korean isolates remain limited. In this study, we conducted a comparative proteomic analysis of the strain TS3128, cultured in three media representing different growth conditions. Proteins related to virulence, type III secretion system, and amylovoran production, were more abundant under minimal conditions than in rich conditions. Additionally, various proteins associated with energy production, carbohydrate metabolism, cell wall/membrane/envelope biogenesis, and ion uptake were identified under minimal conditions. The strain TS3128 expresses these proteins to survive in harsh environments. These findings contribute to understanding the cellular mechanisms driving its adaptations to different environmental conditions and provide proteome profiles as reference for future studies on the virulence and adaptation mechanisms of South Korean strains.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.271-280
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• 2010

Vibrio alginolyticus, a Gram-negative marine bacterium, is one of the causative agents of fish vibriosis. Its virulence factors and pathogenesis mechanism are barely known, except for some extracellular products (ECPs) that are known to be regulated by quorum sensing system. Therefore, the present study used a microarray to analyze the transcription profiles of the wild-type V. alginolyticus and a deletion mutant of luxO, the pivotal regulator in Vibrio quorum sensing systems, which resulted in the identification of a putative virulence factor, MviN. Quantitative real-time reverse transcription PCR confirmed that the transcription of mviN was upregulated in the luxO mutant when compared with wild-type, and down regulated in a luxO-con complemented strain. Furthermore, Western blotting indicated that MviN was greatly induced during the late-exponential and stationary phases of growth, indicating that the expression of MviN was cell-density dependent and quorum sensing regulated in V. alginolyticus. Meanwhile, the mviN null mutant displayed a much slower growth rate than the wild type, signifying the essential role of MviN in V. alginolyticus. Western blotting also revealed that MviN was present as an extracellular protein in V. alginolyticus. When epithelioma papulosum cyprini (EPC) cells were treated with the ECPs of the mviN mutant, no cytotoxicity was observed, whereas EPC cells treated with the wild type exhibited pathological changes, which increased with the ECPs concentration and treatment time. Therefore, the results demonstrated that MviN is a LuxO-regulated ECPs component and involved in the pathogenicity of V. alginolyticus.

• Korean Journal of Veterinary Research
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• v.50 no.1
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• pp.29-35
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• 2010

Yersinia (Y.) ruckeri has been recognized as a serious bacterial pathogen to several kinds of fish, including rainbow trout. However, there are no reports about the characteristics and pathogenicity of Y. ruckeri isolated from farm-cultured eels. In this study, we isolated and characterized Y. ruckeri from the farm-cultured eel Anguilla japonica in Korea. We investigated the phenotypic and genotypic characteristics of Y. ruckeri and tested the virulence of Y. ruckeri isolates on experimentally infected eels. Examination of the flagellar morphology of Y. ruckeri by electron microscopy showed peritrichous flagella in its cell body. Biochemical reaction studies showed overall identical profiles between the isolates and the reference strain of Y. ruckeri in API 20E and API ZYM tests. We sequenced the 16S rRNA of the Y. ruckeri (1,505 bp) for the genotypic characterization (National Center for Biotechnology Information accession number EU401667). Comparison of the 16S rRNA sequences with previously reported Y. ruckeri strains revealed similar phylogenetic relationships. In the virulence assay of the Y. ruckeri on eels, the eels exhibited listlessness, but Y. ruckeri was reisolated from those of the gills and kidneys.

• The Plant Pathology Journal
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• v.37 no.5
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• pp.476-488
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• 2021

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot for cruciferous vegetables worldwide, especially for the cole crops such as cabbage and cauliflower. Due to the lack of resistant cabbage cultivars, black rot has brought about considerable yield losses in recent years in China. Understanding of the pathogen features is a key step for disease prevention, however, the pathogen diversity, population structure, and virulence are largely unknown. In this study, we studied 50 Xcc strains including 39 Xcc isolates collected from cabbage in 20 regions across China, using multilocus sequence genotyping (MLST), repetitive DNA sequence-based PCR (rep-PCR), and pathogenicity tests. For MLST analysis, a total of 12 allelic profiles (AP) were generated, among which the largest AP was AP1 containing 32 strains. Further cluster analysis of rep-PCR divided all strains into 14 DNA groups, with the largest group DNA I comprising of 34 strains, most of which also belonged to AP1. Inoculation tests showed that the representative Xcc strains collected from diverse regions performed differential virulence against three brassica hosts compared with races 1 and 4. Interestingly, these results indicated that AP1/DNA I was not only the main pathotype in China, but also a novel group that differed from the previously reported type races in both genotype and virulence. To our knowledge, this is the first extensive genetic diversity survey for Xcc strains in China, which provides evidence for cabbage resistance breeding and opens the gate for further cabbage-Xcc interaction studies.

• Animal Bioscience
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• v.34 no.4
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• pp.734-742
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• 2021

Objective: The present study aimed to investigate the occurrence and species of coagulase-positive staphylococci (CoPS) and coagulase-negative staphylococci (CoNS) in retail pork meat samples collected during nationwide monitoring. The staphylococcal isolates were characterized for antimicrobial and zinc chloride resistance and enterotoxigenic potential. Methods: A total of 260 pre-packaged pork meat samples were collected from 35 retail markets in 8 provinces in Korea for isolation of staphylococci. Antimicrobial and zinc chloride resistance phenotypes, and genes associated with the resistance phenotypes were determined on the isolates. Furthermore, the presence and distribution of 19 staphylococcal enterotoxin (SE) genes and enterotoxin-like genes among the pork-associated staphylococci were determined by multiplex polymerase chain reaction-based assays using the specific primer sets. Results: A total of 29 staphylococcal strains (29/260, 11.1%) were isolated from samples of retail pork meat, 24 (83%) of which were CoNS. The four CoNS species identified were S. saprophyticus (n = 16, 55%), S. sciuri (n = 3, 10%), S. warneri (n = 3, 10%), and S. epidermidis (n = 2, 7%). Among the 29 isolates, four methicillin-resistant CoNS (MR-CoNS; three S. sciuri and one S. epidermidis) and one methicillin-resistant CoPS (MR-CoPS; one S. aureus) were identified. In addition, a relatively high level of tetracycline (TET) resistance (52%) was confirmed in CoNS, along with a predominant distribution of tet(K). The most prevalent SEs were sep (45%), and sen (28%), which were carried by 81% of S. saprophyticus. Conclusion: These findings suggest that CoNS, especially S. saprophyticus strains, in raw pork meat could be a potential risk factor for staphylococcal food poisoning (SFP), and therefore, requires further investigation to elucidate the role of SEls in SFP and virulence of the pathogen. Our results also suggest that CoNS from raw pork meat may act as a source for transmission of antimicrobial resistance genes such as staphylococcal cassette chromosome mec and tet(K).

• Journal of Microbiology and Biotechnology
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• v.21 no.4
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• pp.366-373
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• 2011

We characterized the proteomes of murine N2a cells following infection with three rabies virus (RV) strains, characterized by distinct virulence phenotypes (i.e., virulent BD06, fixed CVS-11, and attenuated SRV9 strains), and identified 35 changes to protein expression using two-dimensional gel electrophoresis in whole-cell lysates. The annotated functions of these proteins are involved in various cytoskeletal, signal transduction, stress response, and metabolic processes. Specifically, a-enolase, prx-4, vimentin, cytokine-induced apoptosis inhibitor 1 (CIAPIN1) and prx-6 were significantly up-regulated, whereas Trx like-1 and galectin-1 were down-regulated following infection of N2a cells with all three rabies virus strains. However, comparing expressions of all 35 proteins affected between BD06-, CVS-11-, and SRV9-infected cells, specific changes in expression were also observed. The up-regulation of vimentin, CIAPIN1, prx-4, and 14-3-3 ${\theta}/{\delta}$, and down-regulation of NDPK-B and HSP-1 with CVS and SRV9 infection were ${\geq}2$ times greater than with BD06. Meanwhile, Zfp12 protein, splicing factor, and arginine/serine-rich 1 were unaltered in the cells infected with BD06 and CVS-11, but were up-regulated in the group infected with SRV9. The proteomic alterations described here may suggest that these changes to protein expression correlate with the rabies virus' adaptability and virulence in N2a cells, and hence provides new clues as to the response of N2a host cells to rabies virus infections, and may also aid in uncovering new pathways in these cells that are involved in rabies infections. Further characterization of the functions of the affected proteins may contribute to our understanding of the mechanisms of RV infection and pathogenesis.