• Journal of Microbiology and Biotechnology
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• v.20 no.8
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• pp.1163-1178
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• 2010

Plasmids play important roles in horizontal gene transfer among Vibrionaceae, but surprisingly little is known about their replication and incompatibility systems. In this study, we successfully developed a bioinformatics-assisted strategy of experimental identification of seven Vibrio plasmid replicons. Comparative sequences analysis of the seven Vibrio plasmid replicons obtained in this study together with eight published Vibrionaceae plasmid sequences revealed replication-participating elements involved in the ColE1 mode of replication initiation and regulation. Like plasmid ColE1, these Vibrionaceae plasmids encode two RNA species (the primer RNA and the antisense RNA) for replication initiation and regulation, and as a result, the 15 Vibrionaceae plasmids were designated as ColE1-like Vibrionaceae (CLV) plasmids. Two subgroups were obtained for the 15 CLV plasmids, based on comparison of replicon organization and phylogenetic analysis of replication regions. Coexistence of CLV plasmids were demonstrated by direct sequencing analysis and Southern hybridization, strongly suggesting that the incompatibility of CLV plasmids is determined mainly by the RNA I species like the ColE1-like plasmids. Sequences resembling the conserved Xer recombination sites were also identified on the CLV plasmids, indicating that the CLV plasmids probably use the host site-specific recombination system for multimer resolution like that used by ColE1-like plasmids. All the results indicated that the 15 plasmids form a new ColE1-like group, providing a basis for the rapid characterization and classification of Vibrionaceae plasmids.

• Journal of Microbiology and Biotechnology
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• v.30 no.6
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• pp.830-838
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• 2020

The histone-like nucleoid structuring protein (H-NS) is an abundant global regulator of environmentally controlled gene expression. Herein, we demonstrate that H-NS represses the expression of LeuO, the master regulator of the cyclic(Phe-Pro)-dependent signaling pathway, by directly binding to the upstream region of the gene. H-NS binds to a long stretched region (more than 160-bp long), which overlaps with binding sites for ToxR and LeuO. A high quantity of H-NS outcompetes ToxR for binding to the cis-acting element of leuO. However, our footprinting analyses suggests that the binding of H-NS is relatively weaker than LeuO or ToxR at the same molarity. Considering that the DNA nucleotide sequences of the upstream regions of leuO genes are highly conserved among various Vibrio, such patterns as those found in V. vulnificus would be a common feature in the regulation of leuO gene expression in Vibrionaceae. Taken together, these results suggest that, in species belonging to Vibrionaceae, H-NS regulates the expression of leuO as a basal stopper when cFP-ToxR mediated signaling is absent.

• Korean Journal of Food Science and Technology
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• v.20 no.3
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• pp.357-362
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• 1988

Twenty-one aerobic bacteria were isolated from Kimchi and the influences of aqueous extract of garlic on the isolates were studied. The bacteria were 11 Bacillus spp., 2 Staphylococcus spp., a Micrococcus sp., a Flavobacterium sp., an Enterobacteriaceae, and 4 Vibrionaceae. In nutrient broth growth of the bacteria was inhibited by the extract. The antibacterial effects of the garlic were different from each other depending upon the bacteria. The results revealed that the inhibitory effects were due to the bacteriocidal action of the garlic extracts.

• Journal of fish pathology
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• v.32 no.2
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• pp.69-80
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• 2019

We performed MLSA (multilocus sequence analysis) and phenotypic characterization of Aliivibrio species isolated from walleye pollock (Gadus chalcogrammus) maintained in 3 different facilities of Gangwon Province, the east coast of Korea. Of 38 Aliivibrio species identified by 16S rDNA sequences, 12 strains were randomly selected and MLSA was conducted with 5 house-keeping genes (gapA, gyrB, pyrH, recA and rpoA) and 16S rDNA gene. Phylogenetic analysis and homology of the concatenated sequences (4,580 bp) with other Vibrionaceae genera revealed that 4 strains (GNGc16.1, YYGc16.1, YYGc16.2, GSGc18.1) were identified as Aliivibrio logei and one strain (GSGc16.1) as A. wodanis. One strain (GSGc17.1) was tentatively identified as A. logei, but needs further analysis because it did not belong to the same clade with A. logei type strain. 6 strains (GSGc17.2, GNGc16.2, GSGc16.2, GSGc17.3, GSGc18.2, GSGc17.4) need further investigation as potential novel species. Either phenotypic characterization or 16S rDNA sequence alone did not provide enough information for identification of Aliivibrio strains at the species level. A. logei and A. wodanis are generally known as non-pathogenic bacteria, but also known as opportunistic or secondary pathogens of cold water fishes. Cares should be taken to prevent potential outbreaks due to these bacteria, although there was no outbreaks during the sampling period.

• Journal of Life Science
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• v.28 no.3
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• pp.361-368
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• 2018

This study investigated the diversity of communities of intestinal microorganisms, separated from the intestinal organs of Red tilefish (Branchiostegus japonicas), collected on the Jeju Coast. First, in the isolation of 1.5% BHIA, MA, TSA and R2A Agar on the medium, there were most colonies in 1.5% BHIA. The results of aerobic culture and anaerobic culture were $1.7{\times}10^6CFU/g^{-1}$ and $1.1{\times}10^5cfu/g^{-1}$, respectively, on average, and 147 pure colonies were separated in total. In 16S rDNA sequencing, there were 58 genera and 74 species, showing 95-100% similarity with the basic strain. They were divided broadly into 5 phyla, and as the main phyletic group, Proteobacteria phylum comprised 50% with 9 families, 35 genera and 35 species of Moraxellaceae, Rhodobacteraceae, Shewanellae, Halomondaceae, Enterobacteriaceae, Vibrionaceae, Hahellaceae, Pseudomonadaceae, and Erythrobacteraceae, with the highest index of dominance. Actinobacteria phylum comprised 24% with 8 families, 11 genera and 17 species of Microbacteriaceae, Intrasporangiaceae, Dietziaceae, Dermabacteraceae, Dermacoccaceae, Nocardiodaceae, Brevibacteriaceae and Propionobacteriacea; Firmicutes phylum, 16% with 6 families, 8 genera and 17 species of Bacillaceae, Staphylcoccaceae, Planococcaceae, Streptococcaceae, Paenibacillaceae and Clostridiaceae; Bacteroidetes phylum, 6% with 2 families, 3 genera and 4 species of Cyclobacteriaceae and Flavobacteriaceae; and Deinococcus-Thermus phylum, 4% with 1 family, 1 genus and 1 species of Deinococcaceae.

• Journal of Life Science
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• v.28 no.12
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• pp.1477-1488
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• 2018

This study investigated the muscles, intestines, and gonads of Sulculus diversicolor supertexta to examine the diversity of microbial communities within examples collected from the Jeju Coast. Using different media, initial pure isolation in MA, 1% BHIA, and 1% TSA indicated that the muscles, intestines, and gonads supported more communities, respectively. In analysis of relative similarity with 16s rRNA sequencing, 190 pure colonies were isolated, and further analysis with NBLAST identified 71 species, 39 genera, 25 families, and five phyla. Homogeny with the reference strain was 91-100%. Microbial communities in S. supertexta consisted of gamma and alpha Proteobacteria (48%), Actinobacteria (32.5%), Firmicutes (16.9%), Deinococcus-Thermus (1.3%), and Bacteroides (1.3%). In all tissue, Psychrobacter cibarius in Moraxellaceae was dominant. Alteromonadaceae, Enterobacteriaceae, Pasturellaceae, Moraxellaceae, Rhodobacteraceae, Geminicoccaceae, Dietziaceae, Intrasporangiaceae, Microbacteriaceae, Micrococcaceae, Micromonosporaceae, Streptomycetaceae, Aerococcaceae, Bacillaceae, Paenibacillaceae, Planococcaceae, and Staphylcoccaceae were commonly isolated across all tissues, and Flavobacteriaceae, Corynebacteriaceae, Yesiniaceae, Vibrionaceae, Hahellaceae, Pseudomonadaceae were also identified from the intestines. In microbial monitoring of four harmful bacteria, Streptomyces albus (96%) showed antibacterial activity against all four strains, and Agrococcus baldri (99%) and Psychrobacter nivimaris (99%) presented against E. Coli and E. aerogens. In addition, some strains with low homogeny were isolated and further experiments are therefore required, for example to refine the antimicrobial substances including new strain investigations. These additional experiments would aim to establish generic resources for the microbial communities in S. Supertexta and provide basic data for applied microbiological research.

• Journal of Life Science
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• v.29 no.2
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• pp.248-255
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• 2019

This research confirmed the diversity and characterization of gut microorganisms isolated from the intestinal organs of Muraenesox cinereus, collected on the Samcheonpo Coast and Seocheon Coast in South Korea. To isolate strains, Marine agar medium was basically used and cultivated at $37^{\circ}C$ and pH7 for several days aerobically. After single colony isolation, totally 49 pure single-colonies were isolated and phylogenetic analysis was carried out based on the result of 16S rRNA gene DNA sequencing, indicating that isolated strains were divided into 3 phyla, 13 families, 15 genera, 34 species and 49 strains. Proteobacteria phylum, the main phyletic group, comprised 83.7% with 8 families, 8 genera and 26 species of Aeromonadaceae, Pseudoalteromonadaceae, Shewanellaceae, Enterobacteriaceae, Morganellaceae, Moraxellaceae, Pseudomonadaceae, and Vibrionaceae. To confirm whether isolated strain can produce industrially useful enzyme or not, amylase, lipase, and protease enzyme assays were performed individually, showing that 39 strains possessed at least one enzyme activity. Especially the Aeromonas sp. strains showed all enzyme activity tested. This result indicated that isolated strains have shown the possibility of the industrial application. Therefore, this study has contributed for securing domestic genetic resources and the expansion of scientific knowledge of the gut microbial community in Muraenesox cinereus of South Korea.

• Journal of Life Science
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• v.32 no.9
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• pp.721-728
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• 2022

This study investigated the diversity of intestinal microbial communities isolated from the intestine of topshell (Turbo cornutus) from the coast of Jeju Island (Beobhwan, Seogwipo city). Pure cultivation using the standard marine agar (MA) medium showed the most significant number of clusters. Aerobic and anaerobic culture allowed isolation of strains of 1.8×10⁵ CFU·g^-1 and 0.4×10 CFU·g^-1 on average, respectively. The microbial population in the topshell intestine was classified into 4 phyla, 12 families, 26 genera, and 67 species. The microbes in the topshell intestine were detected by homology with 93~100% with standard strains. The microbes in the topshell intestine consisted of Proteobacteria 39%, Firmicutes 34%, Actinobacteria 21%, and Bacteroidets 6%. The identified families were Alteromonadaceae (1), Shewanellaceae (4), Vibrionaceae (12), Phyllobacteriaeceae (1), Rhodobacteraceae (8), Bacillaceae (21), Paenibacillaceae (2), Cellulomonadaceae (1), Mycobacteriaceae (6), Nocardiaceae (4), Streptomycetaceae (3) and Flavobacteriaceae (4). Bacillus sp. and Vibrio sp. accounted for the greatest portion of the separated strains. Among the isolated microorganisms, some strains had probiotic functions.

• Journal of Microbiology
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• v.43 no.1
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• pp.1-10
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• 2005

We isolated and cultured bacteria inhabiting solar saltern ponds in Taean-Gun, Chungnam Province, Korea. All of the isolated 64 strains were found to be moderately halophilic bacteria, growing in a salt range of 2-20 %, with an optimal concentration of 5% salt. Bacterial diversity among the isolated halophiles was evaluated via RFLP analyses of PCR-amplified 16S rDNAs, followed by phylogenetic analysis of the partial 16S rDNA sequences. The combination of restriction enzyme digestions with HaeIII, CfoI, MspI and RsaI generated 54 distinct patterns. A neighbor-joining tree of the partial 16S rDNA sequences resulted in the division of the 64 strains into 2 major groups, 45 strains of ${\gamma}-Proteobacteria$ (70.3%) and 19 strains of Firmicutes (29.7%). The ${\alpha}-Proteobacteria$ and Cytophaga-Flavobacterium-Bacterioides groups, which were repeatedly found to exist in thalassohaline environments, were not represented in our isolates. The ${\gamma}-Proteobacteria$ group consisted of several subgroups of the Vibrionaceae (37.5%), Pseudoalteromonadaceae (10.9%), Halomonadaceae (7.8%), Alteromonadaceae (7.8%), and Idiomarinaceae (6.3%). Members of Salinivibrio costicola (29.7%) were the most predominant species among all of the isolates, followed by Halobacillus treperi (12.5%). Additionally, three new species candidates were found, based on similarities of the 16S rDNA sequences to those of previously published species.

• IMMUNE NETWORK
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• v.8 no.3
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• pp.67-74
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• 2008

Background: The effects of the dietary administration of two heat-inactivated whole bacteria from the Vibrionaceae family, singly or combined, on innate immune response of the rainbow trout were studied. The two bacteria (Pdp11 and 51M6), which were obtained from the skin of rainbow trout, showed in vitro characteristics that suggested they could be considered as potential fish probiotics. Methods: The fish were fed four different diets: control (non-supplemented), or diets supplemented with heat-inactivated bacteria at $10^8$ cfu/g Pdp11, $10^8$ cfu/g 51M6 or with $0.5{\times}10^8$ cfu/g Pdp11 plus $0.5{\times}10^8$ cfu/g 51M6 for 4 weeks. Six fish were sampled at weeks 1, 2, 3 and 4, and then the main humoral (natural haemolytic complement activity and serum peroxidase content) and cellular innate immune responses (leucocyte peroxidase content, phagocytosis, respiratory burst and cytotoxicity) were evaluated. Results: The serum peroxidase content and the natural haemolytic complement activity increased with time, reaching the highest values in the third and fourth weeks of feeding, respectively. The phagocytic ability of specimens fed the mixture of the two inactivated bacteria was significantly higher than in the controls after 2 and 3 weeks of treatment. The same activity increased significantly in rainbow trout fed the Pdp11 diet for 2 weeks or the 51M6 diet for 3 weeks. Respiratory burst activity was unaffected by all the experimental diets at all times assayed. Cytotoxic activity had significantly increased after 3 weeks in fish fed the 51M6 diet. Conclusion: Our results demonstrated the usefulness of incorporating inactivated probiotic bacteria into fish diets.