• Korean Journal of Microbiology
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• v.51 no.1
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• pp.21-30
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• 2015

The goal of this study was to identify relationships between the composition of sulfate reducing bacterial assemblages and terminal restriction fragment length polymorphism (T-RFLP) patterns in rice paddy and dry farming soils. Samples of organic farming soils, conventional farming soils, and dry field farming soils were collected in August and November. Analyses of the soil chemical composition revealed similar total nitrogen, total carbon and total inorganic phosphorus levels; however, the moisture content and total carbon were higher than in the other soils in both August and November, respectively. Sulfate reducing bacteria utilizing lactic acid were more widely distributed than those that used acetic acid, and the number of sulfate reducing bacteria in organic farming soil was most abundant. Phylogenetic analysis based on 181 clones revealed that most showed low similarity with cultured sulfate reducing bacteria, but more than 90% similarity with an uncultured sulfate reducing bacteria isolated from the environment. T-RFLP analysis revealed that fragments of 91, 357, 395, and 474 bp were most common, and the community structure of sulfate reducing bacteria changed seasonally.

• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.611-615
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• 2008

Novel groups of uncultivable anaerobic thermophiles were isolated from compost by enrichment cultivation in medium with a cell-free extract of Geobacillus toebii. The cell-free extract of G. toebii provided the medium with growth-supporting factors (GSF) needed to cultivate the previously uncultured microorganisms. Twenty-nine GSF-requiring candidates were successfully cultivated, and 16 isolated novel bacterial strains were classified into three different groups of uncultivable bacteria. The similarity among these 16 isolates and a phylogenetic analysis using 16S rRNA gene sequences revealed that these GSF-requiring strains represented novel groups within the family Clostridiaceae.

• Korean Journal of Environmental Biology
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• v.29 no.4
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• pp.265-273
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• 2011

Bacterial diversity based on the denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA gene sequences was determined for soil samples from two abandoned mine sites and the corresponding enrichment cultures using soil sample as key inoculum. Sequencing analysis of DGGE bands obtained from both the soil samples matched mostly with sequences of uncultured and newly described organisms, or organisms recently associated with the acid mine drainage environment. However, the enrichment of soil samples in ferrous sulfate and elemental sulfur media yielded sequences that were consistent with well-known iron- and sulfur-oxidizing acidophilic bacteria. Analysis of enrichment cultures of soil samples from Dalsung mine revealed abundant ${\gamma}$-$Proteobacteria$, whereas that of Gubong mine sample displayed acidophilic groups of ${\gamma}$-$Proteobacteria$, ${\alpha}$-$Proteobacteria$, $Actinobacteria$ and $Firmicutes$. Chemical elemental analysis of the mine samples indicated that the Dalsung site contained more iron and sulfate along with other toxic components as compared with those of the Gubong site. Biogeochemistry was believed to be the primary control on the acidophilic bacterial group in the enrichment samples.

• Journal of Microbiology and Biotechnology
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• v.26 no.5
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• pp.883-890
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• 2016

Marine sediments are a microbial biosphere with an unknown physiology, and the sediments harbor numerous distinct phylogenetic lineages of Bacteria and Archaea that are at present uncultured. In this study, the structure of the archaeal and bacterial communities was investigated in the surface and subsurface sediments of Jeju Island using a next-generation sequencing method. The microbial communities in the surface sediments were distinct from those in the subsurface sediments; the relative abundance of sequences for Thaumarchaeota, Actinobacteria, Bacteroides, Alphaproteobacteria, and Gammaproteobacteria were higher in the surface than subsurface sediments, whereas the sequences for Euryarchaeota, Acidobacteria, Firmicutes, and Deltaproteobacteria were relatively more abundant in the subsurface than surface sediments. This study presents detailed characterization of the spatial distribution of benthic microbial communities of Jeju Island and provides fundamental information on the potential interactions mediated by microorganisms with the different biogeochemical cycles in coastal sediments.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.4
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• pp.447-453
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• 2015

We determined the optimal culture conditions for obtaining the maximum number of intestinal bacteria from the olive flounder Paralichthys olivaceus, and studied bacterial diversity using both culture-dependent and culture-independent methods. Using six culture conditions, mean bacterial numbers were greater than $10^6$ per gram of gut mucus, regardless of the medium. However, the bacterial diversity, based on colony morphology, appeared much higher on Marine agar (MA) and Zobell 2216 agar than on other media. We found eight and 17 cultured bacterial phylotypes with 99% minimum similarity in gut mucus grown on MA and tryptic soy agar, respectively. Furthermore, we used genomic DNA extracted from gut mucus to generate 78 random clones, which were grouped into 25 phylotypes. Of these, six were affiliated with Firmicutes, Actinobacteria, and Verrucomicrobia, and were not found using our culture-dependent methods. Consequently, we believe that Marine agar and Zobell 2216 agar are optimal media for culturing diverse intestinal microbes; we also discovered several novel sequences not previously recognized as part of the gut microbiota of olive flounder.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.312-317
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• 2015

Pyrosequencing of 16S rDNA tags was used to obtain the bacterial diversity and community structure in the uncultured seawaters as well as in the cultured seawaters, which were collected from the 7 ports (Cheonbu, Hyunpo, Taeha, Namyang, Sadong, Dodong, and Jeodong) and 1 seashore (Guam) around the Ulleng island, Korea. Alphaproteobacteria were the most abundant group in the clean seawaters such as seawaters of Taeha and Sadong ports. Gammaproteobacteria proportion increased depending upon the wastewater amounts mixed with the seawaters such as seawaters of Namyang, Dodong, and Jeodong ports. The genuses of Alteromonas (from samples of Cheonbu, Taeha, Guam, Namyang, Sadong), Shewanella (from sample of Jeodong), and Vibrio (from samples of Hyunpo and Dodong) were dominant group in each of the cultured seawaters incubated in marine broth (Difoco). The results suggest that the incoming wastewaters to the port seawaters contribute to the dynamic change of the marine bacterial community around the Ulleng island.

• Proceedings of the Microbiological Society of Korea Conference
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• 2005.05a
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• pp.28-32
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• 2005

Epifluorescence microscopy and direct viable counting methods have shown that only 0.01-0.1% of all the microbial cells from marine environments form colonies on standard agar plates. To culture novel marine microorganisms, high throughput culturing (HTC) techniques were developed to isolate cells in very low nutrient media. This approaches was designed to address microbial metabolic precesses that occur at natural substrate concentrations and cell densities, which are typically about three orders of magnitude less than in common laboratory media. Approximately 5000 cultures of pelagic marine bacteria were examined over the course of 3 years. Up to 14% of cells from coastal seawater were cultured using this method, a number that is 1400 to 140-fold higher than obtained by traditional microbiological culturing techniques. Among the cultured organisms are many unique phylogenetic lineages that have been named as new phyla (7), orders (2, 5, 12), families (3), and genera (1, 4, 6). Over 90% of the cells recovered by this method do not replicate in standard agar plating, the most common method of microbial cell cultivation.

• Journal of Microbiology and Biotechnology
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• v.22 no.6
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• pp.771-779
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• 2012

The bacterial diversity of the continental shelf sediment in the Yellow Sea was investigated by the cloning and sequencing of PCR-amplified 16S rRNA genes. The majority of the cloned sequences were distinct phylotypes that were novel at the species level. The richness estimator indicated that the sediment sample might harbor up to 32 phylum-level taxa. A large number of low-abundance, phylum-level taxa accounted for most of the observed phylogenetic diversity at our study site, suggesting that these low-abundance taxa might play crucial roles in the shelf sediment ecosystem.

• Journal of Life Science
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• v.15 no.5 s.72
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• pp.743-748
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• 2005

Histamine can be produced at early spoilage stage through decarboxylation of histidine in red-flesh fish by Proteus morganii, Hafnia alvei or Klebsiella pneumoniae. Allergic food poisoning is resulted from the histamine produced when the freshness of Mackerel degrades. Conversely it has been reported that there are bacteria which decompose histamine at the later stage. We isolated histamine decomposers from salted mackerel and studied the characteristics to help establish hygienic measure to prevent outbreak of salted mackerel food poisoning. All the samples were purchased through local supermarket. Histamine decomposers were isolated using restriction medium using histamine 10 species were selected. Identification of these isolates were carried out by the comparison of 16S rDNA partial sequence; as a result, we identified Pseudomonas putida strain RA2 and Halomonas marina, Uncultured Arctic sea ice bacterium clone ARKXV1/2-136, Halomonas venusta, Psychrobacter sp. HS5323, Pseudomonas putida KT2440, Rhodococcus erythropolis, Klebsiella terrigena (Raoultella terrigena), Alteromonadaceae bacterium T1, Shewanella massilia with homology of $100\%,{\;}100\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}100\%,{\;}95\%,{\;}99\%,{\;}and{\;}100\%$respectively. Turbidometry determination method and enzymic method were employed to determine the ability of histamine decomposition. Among those species Shewanella massilia showed the highest in ability of histamine decomposition. From these results we confirmed various histamine decomposer were present in salted mackerel product in the market.

• Korean Journal of Microbiology
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• v.48 no.4
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• pp.275-283
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• 2012

Culture-dependent RFLP and culture-independent DGGE were employed to investigate the bacterial community associated with the marine sponge Asteropus simplex collected from Jeju Island. A total of 120 bacterial strains associated with the sponge were cultivated using modified Zobell and MA media. PCR amplicons of the 16S rDNA from the bacterial strains were digested with the restriction enzymes HaeIII and MspI, and then assigned into different groups according to their restriction patterns. The 16S rDNA sequences derived from RFLP patterns showed more than 94% similarities compared with known bacterial species, and the isolates belonged to five phyla, Alphaproteobacteria, Gammaproteobacteria Actinobacteria, Bacteroidetes, and Firmicutes, of which Gammaproteobacteria was dominant. DGGE fingerprinting of 16S rDNAs amplified from the sponge-derived total gDNA showed 12 DGGE bands, and their sequences showed more than 90% similarities compared with available sequences. The sequences derived from DGGE bands revealed high similarity with the uncultured bacterial clones. DGGE revealed that bacterial community consisted of seven phyla, including Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Actinobacteira, Chloroflexi, and Nitrospira. Alphaproteobacteria, Gammaproteobacteria, and Actinobacteria were commonly found in bacteria associated with A. simplex by both RFLP and DGGE methods, however, overall bacterial community in the sponge differed depending on the analysis methods. Sponge showed more various bacterial community structures in culture-independent method than in culture-dependent method.