• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.207-215
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• 2010

The purpose of this research is to draw the bacterial community difference between Korean and Chinese kimchi for future use in the confirmation of kimchi origin. Initial fermentation stage kimchi samples (above pH 5) were used for the analysis of bacterial diversity. From 26 Korean kimchi samples, 1,017 strains in the 45 genera and from 22 Chinese kimchi samples, 842 strains in the 54 genera were isolated with use of marine medium, nutrient medium, succinate minimal medium (SMM), leuconostocs selective medium (LUSM) agars. In the order of isolated numbers, Bacillus, Weissella, Leuconostoc, Pseudomonas, and Lactobacillus genera and Bacillus, Weissella, Lactobacillus, Pseudomonas, Serratia, and Enterobacter genera were predominated in Korean and Chines kimchi, respectively. Among the isolated lactic acid bacteria, Weissella spp. were isolated most dominantly owing to the biased growth of Weissella spp. on LUSM agar. Species in the genera Leuconostoc and Lactobacillus were the next frequently isolated LAB from Korean and Chinese kimchi, respectively. Weissella confusa was isolated only from Korean kimchi and W. soli and Serratia proteamculans were isolated only from Chinese kimchi. They have a possibility to be used as target bacteria to differentiate Korean kimchi from Chinese kimchi.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1621-1628
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• 2012

The agar-degrading bacterium GNUM-1 was isolated from the brown algal species Sargassum serratifolium, which was obtained from the West Sea of Korea, by using the selective artificial seawater agar plate. The cells were Gram-negative, $0.5-0.6{\mu}m$ wide and $2.0-2.5{\mu}m$ long curved rods with a single polar flagellum, forming nonpigmented, circular, smooth colonies. Cells grew at $20^{\circ}C-37^{\circ}C$, between pH 5.0 and 9.0, and at 1-10% (w/v) NaCl. The DNA G+C content of the GNUM-1 strain was 45.5 mol%. The 16S rRNA sequence of the GNUM-1 was very similar to those of Alteromonas stellipolaris LMG 21861 (99.86% sequence homology) and Alteromonas addita $R10SW13^T$(99.64% sequence homology), which led us to assign it to the genus Alteromonas. It showed positive activities for agarase, amylase, gelatinase, alkaline phosphatase, esterase (C8), lipase (C14), leucine arylamidase, valine arylamidase, ${\alpha}$-chymotrypsin, acid phosphatase, naphthol-AS-BI-phosphohydrolase, ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\beta}$-glucosidase, catalase, and urease. It can utilize citrate, malic acid, and trisodium citrate. The major fatty acids were summed feature 3 (21.5%, comprising $C_{16:1}{\omega}7c/iso-C_{15:0}$ 2-OH) and C16:0 (15.04%). On the basis of the variations in many biochemical characteristics, GNUM-1 was considered as unique and thus was named Alteromonas sp. GNUM-1. It produced the highest agarase activity in modified ASW medium containing 0.4% sucrose, but lower activity in rich media despite superior growth, implying that agarase production is tightly regulated and repressed in a rich nutrient condition. The 30 kDa protein with agarase activity was identified by zymography, and this report serves as the very first account of such a protein in the genus Alteromonas.

• Journal of the korean academy of Pediatric Dentistry
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• v.37 no.2
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• pp.186-192
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• 2010

Streptococcus mutans, one of the major causal agents of dental caries, is component of the dental plaque. It produces various organic acids such as lactic acid which is the end-product of glycolysis, and this leads to dental caries. A new system using species-specific monoclonal antibodies was developed to detect Streptococcus mutans in saliva. The system quickly detects salivary Streptococcus mutans in 30min and classifies the result into two levels. The purpose of this study was to investigate correlation between monoclonal antibody-based detecting system and selective medium-based detecting methods. Children's deft indices were also compared with Streptococcus mutans counts in MSB agar plate. Subjects consisted of 15 children in the age of 3 to 6 years. They were assigned to three groups : Group I(deft index = 3), Group II(deft index $\leqq$3), Group III(deft index $\geqq$4). The results are as follows : 1. The rate of children with positive response was 13.3% and with negative response was 86.7% in the result of Saliva-checkTM Mutans test kit. 2. There was a positive correlation between monoclonal antibody-based detecting system and selective medium-based detecting methods(p<0.05). 3. Streptococcus mutans counts in MSB agar plate were irrelevant to deft of children(p=0.34).

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.79-85
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• 2002

Ecological characteristics of microorganisms in tideland soils were studied by investigation of microbial diversity and population. Twenty soil samples were taken at surface, 10, 20 and 30 cm depth each. Bacteria, actinomycetes and fungi were isolated on each selective isolation medium containing different concentration of NaCl. Actinomycetes were the most isolated from soil samples taken at 10 cm depth and isolated by humic acid-vitamin (HV) medium without sea water or salt. Twenty nine strains of actinomycetes were isolated at surface soil and 74, 39, 37 strains were at 10, 20, and 30 cm depth, respectively. All these isolates were analysed and grouped by random amplified polymorphic DNA (RAPD)-PCR analysis. Many of the isolates were clustered into Microtetraspora and Pseudonocardia. Fungal isolates were highly distributed at the surface soil and isolated well on potato dextrose agar (PDA) medium with sea water. Bacterial isolates were higly distributed at surface soil and isolated well by nutrient medium without sea water or salt. Soil samples taken at 10 cm depth showed the highest microbial diversity and population.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.3
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• pp.367-376
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• 1997

Vibrio vulnificus is a gram-negative, halophilic, oxidase-positive, lactose-positive, motile, rod shaped bacterium that has been associated with primary septicemia and wound infection. Elucidating the growth and survival of V. vulnificus in ecological conditions is of great importance to develop sanitary measure against this microorganism. Thus we simulated the ecological conditions and evaluated the effect. About $10^5\;CFU/ml$ of V. vulnificus was inoculated to fresh water, brackish water $(1\%\;NaCl)$, sea water $(3\%\;NaCl)$, and bottom deposit solution. The same concentration of V. vulnificus was also inoculated to distilled water, $1\%\;NaCl$ solution and $3\%\;NaCl$ solution as controls. These were stored at 4, 15 and $25^{\circ}C$, respectively and were used to assess the effects of temperature and salinity on the survival of V. vulnificus. In fresh water V. vulnificus could not survive regardless of storage temperature. In case of brackish water and sea water survival time of V. vulnificus was the longest at $25^{\circ}C$, and the number of V. vulnificus was decreased most rapidly at $4^{\circ}C$. V. vulnificus survived longer in brackish water than in any other conditions. In bottom deposit solution containing brackish water, the survival time of V. vulnificus was longer and the rate of decline was slower than that in brackish water. These results indicate that both biological and physicochemical factors such as temperature and salinity could affect survival of V. vulnificus. V. vulnificus, damaged in normal fresh water, did not grow on TCBS agar of selective plating medium but grew on BHI agar plate; However, V. vulnificus was recovered by addition of salt and nutrient materials.

• KSBB Journal
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• v.13 no.2
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• pp.214-219
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• 1998

Polymerase chain reaction(PCR) was conducted to obtain the gene encoding glucose oxidase(GOD) from Aspergillus niger(ATCC 2110) and the DNA sequence determined was coincided with published GOD sequence from A. niger. Recombinant transforming vector containing GOD and hygromycin B(hyg.B) resistant gene(hph) was constructed and used for further transformation of A. niger ATCC 2110. Selectivity of hyg.B against A. niger differed depending on which media were used i.e., nutrient-rich media such as potato dextrose agar(PDA) and complete medium(CM) showed only 50% growth inhibition at 400 $\mu$m ml$^-1$ of hyg.B while the minimal media inhibited mycelial growth completely at 200 $\mu$m ml$^-1$ of hyg.B. Twenty to sixty putative transformants were isolated from the hyg.B-containing minimal top agar, transferred successively onto alternating selective and nonselective media for a mitotic stability of hyg.B resistance and, then, single-spored. Among the stable transformants, the transformant(GOD1-6) grown by flask culture showed the considerable increase of extracellular GOD activity, which was estimated to the degree of 50% - 100% comparing to that of wild type. Transformation of tGOD1-6 was resulted from integration of the vectors into heterologous as well as homologous regions of the A. niger genome. Southern blot analysis revealed that there were two independent integrations of vector into fungal genome and one into the GOD gene due to homologous recombination. In addition, GOD activity and sodium gluconate production when tGOD1-6 was fed-batch fermented were enhanced 11 fold and 2.25 fold, respectively, compared to that of the wild type.

• Journal of Periodontal and Implant Science
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• v.33 no.2
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• pp.149-158
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• 2003

The purpose of this study was to isolate and characterize the Fusohacrerium nucleatum (F. nucleatum) from subgingival plaque in Korean periodontitis patients. The subgingival plaque samples of periodontitis patient were collected with sterilized paper point. The paper point was put into reduced transfer medium and then immediately transferred to laboratory. The subgingival samples were diluted by 10,000 folds and plated on F. nucleatum-selective media agar plate. The plates were incubated at 37$^{\circ}C$ in an anaerobic chamber for 3 days. The violet-colored colonies were selected and subjected to further verification whether those are F. nucleatum or not. For further confirmation, 16S rRNA genes (rDNA) were cloned from each of bacterial clones and determined sequence of 16S rDNA. In this study, we found 17 distinct clinical isolates of F. nucleatum from subgingival plaque. The clinical isolates will be a useful in various studies in periodontology.

• The Plant Pathology Journal
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• v.25 no.2
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• pp.121-127
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• 2009

Since July 2005, survey of chestnut ink disease was carried out in chestnut stands located at southern parts of Korea. Dead chestnut trees showing inky ooze on necrotic trunks were found in two different locations. In order to isolate and identify the causal fungus, infected tissues and soil samples around dead or dying trees were collected and placed on Phytophthora-selective medium. Rhododendron and chestnut tree leaves were used as a bait to isolate the fungus from soil samples by attracting zoospores in soil suspensions. On V-8 culture medium, the isolates produced homothallic oogonia with protuberances ($34.0-46.2{\times}21.9-26.7{\mu}m$) abundantly, but did not produced sporangia. Mass production of sporangia was possible by immersing agar plugs with actively growing mycelium in the creek water at $18^{\circ}C$ for 3 days. Sporangia were papillate, and ovoid to obpyriform ($17.0-38.9{\times}14.6-29.2{\mu}m$) in shape. Comparison of the ITS sequences revealed that the isolates had 100% identity to the P. katsurae isolates from Japan and New Zealand and 99.6% identity to other P. katsurae isolates. All of the examined isolates from Korea were completely identical to each other in ITS sequence. Numerous sporangia were formed in filtered as well as unfiltered creek water, but no sporangia formed in sterilized distilled water. Light induced sporangia formation, but has no influence on oospore formation. Amendments of ${\beta}$-sitosterol in culture media have no significant effect on mycelial growth but significantly stimulate oospore and sporangia formation.

• Archives of Pharmacal Research
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• v.28 no.6
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• pp.660-666
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• 2005

The intestinal microbiota are important to the host with regard to resistance they impart against bacterial infections and their involvement in mediating metabolic functions. Lactic acid producing bacteria such as Lactobacillus play an important physiological role in these matters. The aim of the present study was to isolate Lactobacillus sp. that inhibits enteric pathogens. Initially, 17 isolates from healthy Koreans were collected on Lactobacillus selective medium. Resistance of the isolates to antibiotics including rifampicin, streptomycin, clindamycin and vancomycin was measured. One of the isolate was identified as Lactobacillus ruminus on the basis of bacterial cell morphology, cultural characteristic and biochemical characteristics, 16S rRNA sequence analysis and PCR-RAPD. Antimicrobial activity of the bacterium against Vancomycin Intermediate Resistant Staphylococcus aureus (VISA) and Vancomycin-Resistant Enterococci (VRE) was measured. About $10^4$ cells of VISA or VRE were mixed with 1, 5, and 9 mL of L. ruminus SPM 0211 and the final volume was adjusted to 10 mL with brain heart infusion (BHI) broth. The cell suspension was incubated for 3, 6, 9, and 24 h, serially diluted and then plated on BHI agar plates. As numbers of L. ruminus SPM 0211 were increased, viable cell count of VISA and VRE decreased. The strongest antimicrobial activity of SPM 0211 was observed after 9 h incubation in any mixture, almost completely inhibiting the growth of these two bacteria. The results suggest that the freshly isolated L. ruminus SPM 0211 may be used as a pro-biotic microbe that prevents the colonization of enteric pathogens and can thereby promote good gastrointestinal health.

• Journal of Periodontal and Implant Science
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• v.25 no.3
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• pp.487-496
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• 1995

Previous studies have demonstrated that not all A. actinomycetemcomitans produced significant level of leukotoxic factor and its leukotoxicity have associated with serotype and genetic variation. Our aim was to investigate on the interrelationship between serotype and leukotoxicity of an A. actinomycetemcomitans consisting of 13 clinically well characterized. Korean isolates and to evaluate if particular virulent clonal types of A. actinomycetemcomitans are associated with periodontal disease. For this study, 13 strains of A. actinomycetemcomitans from 6 patients with periodontal disease were isolated and identified by using a selective medium(tryptic soy agar supplemented with 10% serum, $75{\mu}g$ of bacitracin and $5{\mu}g$ of vancomycin per ml) in 10% C02 incubator for 3days with routine Gram staining, colony morphology and biochemical test..For serotyping, antisera were prepared from reference strains of 5 serotypes. (ATCC 29523,Y4, SUNY aB 67, IDH 781, IDH 1705) and then ammonium sulfate precipitation, immunoabsorption and indirect immunofluoroscent procedures were done. For analysis of leukotoxicity, sonic extract of A. actinomycetemcomitans exposed to PMN, and trypan blue was stained for counting the cell viability. Finally Southern blot analyses of genomic DNA digested with the restriction enzyme Tag I was done and the Southern blots were hybridized with the 530bp fragment, termed delta 530, originating from the ltx promoter of strain 652 and deleted from strain JP2. Also ltxA-3.1 and SC2 probe from strain JP2 were hybridized with genomic DNA fragments. Results reveal that strains isolated showed approximately equal proportions of 3 serotypes(b, d, e) and serotype b was not detected. 2 patients harbored 2 different serotypes in the same disease site. The prevalence of leukotoxic strain was 23% and there was no relationship between serotype, leukotoxicity and clinical observations. Especially virulent clonal types of Actinobacillus actinomycetemcomitan (JP2 strain) could not found. Further studies are necessary on the genetic polymorphism of leukotoxin and its relations to clinical status.