• Applied Biological Chemistry
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• v.50 no.4
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• pp.268-275
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• 2007

Diversity of the mud flat microbial population in Suncheon Bay was investigated by studying extracellular enzyme activities and 16S rDNA sequences. Four culturable bacterial strains with CMCase, xylanase and protease activities were isolated from the wetland and the mud flat. All the strains produced more xylanase activity than CMCase or protease activity, and the properties of the isolate enzymes from the wetland were similar to those from the mud flat. About 2,000 clones were obtained with the 16S rDNA amplified from the metagenomic DNA isolated from the mud samples. Based on the restriction pattern(s), seventeen clones were selected for base sequence analysis. Of the 17 clones, only 35% (6 clones) were found to be cultured strains and 65% (11 clones) to be uncultured strains. The similarities in the base sequences of the clones ranged from 91.0% to 99.9% with an average similarity of 97.3%. The clones could be divided into 7 groups, Proteobacteria (9 clones, 52.9%), Firmicutes (3 clones, 17.6%), Bacteroidetes (1 clone), Flavobacteria (1 clone), Verrucomicrobia (1 clone), Acidobacteria (1 clone), and Chloroflexi (1 clone). Most of the Proteobacteria clones were gamma Proteobacteria associated with oxidation-reduction of sulfur.

• The Journal of the Korean Society for Microbiology
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• v.15 no.1
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• pp.19-32
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• 1980

Besides the benefits of antimicrobial agents in the control of various infectious diseases, widespread and prolonged use of particular antimicrobial agents has brought about the increase of drug-resistant strains in a community and the profound changes in the pattern of infectious diseases. In Korea, there are some remote villages where no clinics and drug stores are available and the residents in those areas are assumed to have fewer chances to contact with antimicrobial agents. In the present study, the differences in susceptibilities to 14 antimicrobial agents between the isolates from rural areas(R) and Seoul National University Hospital(SNUH, H) were studied. The isolates and their numbers were Staphylococcus aureus, R;55, H;68), Enterococci(R;28, H;30), Escherichia coli(R;40, H;40), Enterobacter aerogenes(R;25, H;21) and Klebsiella pneunoniae(R;58, H;67). Minimal inhibitory concentrations(MIC's) of penicillin, ampicillin, carbenicillin, cephalexin, tetracycline, oxytetracyline, doxycycline, minocycline, gentamicin, kanamycin, streptomycin, erythromycin, troleandomycin and co-trimoxazole were determined by agar dilution method. I. Comparison of MIC's and resistant strain proportions between isolates from SNUH and rural areas. MIC's and/or resistant strain proportions of the isolates from SNUH were significantly higher than those of the isolates from rural areas in the cases of 1. S. aureus to doxycycline, streptomycin and kanamycin. 2. E. coli to penicillin, ampicillin, carbenicillin, tetracycline, oxytetracycline, doxycycline, minocycline, streptomycin, kanamycin, erythromycin and co-trimoxazole. 3. E. aerogences to carbenicillin, tetracycline, oxytetracycline, doxycycline, minocycline, streptomycin, kanamycin, genaamicin and co-trimoxazole. 4. K pneunoniae to penicillin, ampicillin, tetracycline, oxytetracycline, doxycycline, monocycline, streptomycin, kanamycine, gentamicin and co-trimoxazole. However, the mean MIC and resistant strain proportion of S. aureus to tetracycline were higher in isolates from rural areas than in those from SNUH and Enterococci showed no differences in susceptibilities to the antimicrobial agents between isolates from rural areas and from SNUH. Therefore, in general, differenes in susceptibility to these antimicrobial agents between the isolates from rural areas and SNUH were remarkably greater and broader in gram negative enteric bacteria. II. Multiple drug resistance pattern. Patterns and incidences of multiple drug resistance were studied with penicillin, ampicillin, tetracycline, cephalexin, gentamicin, streptomcin, kanamycin and co-trimoxazole in Enterococci, E. coli, E. aeroges and K. pneumoniae. There appeared significant differences in the incidence of multiply drug-resistant strains and multiple drug resistance patterns between the isolates from SNUH and rural areas in Enterococci, E. coli, E. aerogenes and K. pneumoniae. However, there was no difference in the incidence of multiply drug-resistant strains between isolates of S. aureus from SNUH and rural areas but the pattern of multiple resistance of the SNUH strains of S. aureus was diverse, while that of the rural strains was predominantly confined to penicillin-tetracycline combination. The incidence of multigly drug-resistant strains and diversity of their patterns were the highest in E. coli strains isolated from SNUH and there were no multiply drug resistant strrains in Enterococci and K. pneumoniae strains isolated from rural areas. The number of drug-resistance determinants was also different between the isolates from rural areas and SNUH. Most of the multiply drug-resistant strains of E. coli, E. aerogenes and K. pneumoniae isolated from SNUH were resistant to more than 3 kinds of antimicrobial agents, most frequently to ampicillin, tetracycline and streptomycin, while multiply drug-resistant strains from rural areas were resistant to 2 kinds of antimicrobial agents among ampicillin, tetracycline and streptomycin. With drug-resistant E. coli strains, resistance to tetracycline which was used most widely since 1951 was most frequently involved as a part of mutliple drug-resistance, followed by resistance to ampicillin and streptomycin. This strongly suggests that emergence of drug-restant strains in a community is directly dependent on the selective pressure exerted by the antimicrobial agent used. III. Cross resistance. Cross resistance of bacteria was studied among tetracycline penicillin, aminoglycoside and macrolide derivatives by analyzing correlation coefficients of sucseptibilities using the least square method. In this study, there were high correlations among the susceptibilities to related derivatives. It appears that the relatively low correlations in susceptibilities present in some cases are due to intrinsic resistance of E. aerogenes to penicillin, Enterococci to aminoglycoside and E. coli E. aerogenes and K. pneumoniae to macrolide derivatives.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.8
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• pp.1131-1140
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• 2014

This research focused on the effects of different doses of Bacillus subtilis KN-42 on the growth performance, diarrhea incidence, faecal bacterial flora, and the relative number of Lactobacillus and Escherichia coli in faeces of weaned piglets to determine whether the strain can serve as a candidate antimicrobial growth promoter. A total of 360 piglets (initial body weight $7.14{\pm}0.63$ kg) weaned at $26{\pm}2$ days of age were randomly allotted to 5 treatment groups (4 pens per treatment with 18 pigs per pen) for a 28-day trial. Dietary treatments were basal diet without any antimicrobial (negative control; NC), basal diet supplemented with 120 mg/kg feed of neomycin sulfate (positive control; PC) and basal diet supplemented with $2{\times}10^9$ (L), $4{\times}10^9$ (M) and $20{\times}10^9$ (H) CFU/kg feed of B. subtilis KN-42. During the overall period, average daily gain and feed efficiency of piglets were higher in groups PC, M, and H than those in group NC (p<0.05), and all probiotics and antibiotics groups had a lower diarrhea index than group NC (p<0.05). The 16S rDNA gene-based methods were used to analyze faecal bacterial flora on day 28 of experiment. The result of denaturing gradient gel electrophoresis analysis showed that supplementation of B. subtilis KN-42 to the diet changed the bacterial communities, with a higher bacterial diversity and band number in group M than in the other four groups. Real-time polymerase chain reaction analysis showed that the relative number of Lactobacillus were higher in groups PC and H than in group NC (p<0.05), and the supplemented B. subtilis KN-42 to the diet also reduced the relative number of E. coli (p<0.05). These results suggest that dietary addition of B. subtilis KN-42 can improve the growth performance and gastrointestinal health of piglets.

• Korean Journal of Environmental Biology
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• v.29 no.3
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• pp.144-153
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• 2011

In this study, samples were collected from the Sumummulbangdui wetland at the Halla Mountain in Jeju Island in order to isolate novel bacterial strain. Bacterial strains belonging to the class Alphaproteobacteria, and Gammaproteobacteria were isolated after spreading samples onto solid agar media. The 16S rRNA gene sequences of the strains assigned to the two classes were compared to those of type strains of the species. The strains that showed less than 98.7% 16S rRNA gene sequence similarity to the validly published species were considered to be novel species candidates. A total of 19 strains were regarded as novel strains which can be regarded as novel species candidates. In the Alphaproteobacteria, 6 novel strains were affiliated with the genera Novosphingobium, and Rhizobium. A total of 13 novel strains belong to Gammaproteobacteria that assigned to the family Moraxellaceae, Pseudomonadaceae, and Enterobacteriaceae were identified. Cultural, physiological, chemotaxonomic characteristics and fatty acids compositions have been determined for the novel species candidates, and the characteristics are described in this study.

• Korean Journal of Microbiology
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• v.53 no.3
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• pp.156-162
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• 2017

Legionnaires' disease (LD) is a severe and potentially fatal pneumonia caused by colonization of human-made water system and subsequent aerosolization and inhalation of Legionella bacteria. A total of 147 Legionella strains was isolated from environmental water sources from public facilities in Gyeonggi-do, South Korea. The distribution of Legionella isolates was investigated according to facility type, and sample type. L. pneumophila was distributed broadly throughout Gyeonggi-do, accounting for 85.7% of the isolates, and L. pneumophila serogroup (sg) 1 predominated in all of the public facilities. L. wadsworthii predominated among non-L. pneumophila species. We performed comparative analyses of L. pneumophila sg 1 isolated from environment water of public facilities in Gyeonggi-do by pulsed field gel electrophoresis (PFGE) and sequence-based typing (SBT). Thirty-two isolates were classified into 22 types by PFGE and 9 sequence types (STs) by SBT and categorized into 3 groups. ST1 was the most prevalent sequence type and two STs obtained in this study had unique allelic profiles. The use of SBT data from different countries for epidemiology study of LD constitutes a technically uncomplicated and relatively easy method for strain subtyping, especially compared to other contemporary techniques.

• Journal of the Korean Institute of Landscape Architecture
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• v.27 no.3
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• pp.1-17
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• 1999

The purpose of this study is to evaluate the creation techniques of artificial wetland, one of biotopes developed to promote biodiversity in urban areas, and to look for improvement steps. Specifically, artificial wetland creation techniques were categorized into living environment and living creature classification. Being living conditions for creations, habitat environment was reviewed with a focus on water and soil environments. Living creatures were classified into plants, insects, fish, and birds. The evaluation of creation techniques was done in post-construction evaluation while considering the creation of habitats for living creatures. Intervention by users, changes in living environment and living species, and relevance of creation techniques were reviewed. Key results of this study are as follows. (1) Water environment for the living environment of creatures provides a suitable environment conditions for the living of creatures through a process easing the use of piped water. Various water depths and embankment appear to have a positive impact on the living of aquatic life. In particular, embankment covered in soil naturally played an important role as a place for the activities of aquatic insects and young fish as well as the growth of aquatic plants. (2) Various aquatic and ground plants to promote insect-diversity, shallow water, and old-tree logs had contributed greatly in increasing the types and number of insects. Aquatic insects. Aquatic insects were seen much particularly in areas where aquatic plants are rich but water is shallow than any other areas. (3) A space piled with stone to provide habitats for fish was not much used. However, it was observed that fish used embankment built with natural stones and embankment using logs in areas where water is deep. In addition, it was confirmed that 1,500 fish that had been released propagated using various depths and places for birth. (4) It was analyzed that techniques (creation of island, log setting, and creation of man-made bird nests) to provide habitats and to attract birds are not serving their roles. In such a case, it is believed that species had not increased due to the smallness as well as isolated features of the area. Based on theoretical review, they are judged to be areas that are likely to be used when a greater variety of birds is introduced. It is judged that attracting and keeping more birds at the site, such spaces need to be linked systematically in the future in terms of building eco-network while ensuring an adequate living areas. (5) In the study areas, users intervened greatly. As a result, a blockage was created preventing the normal growth of plants and non-indigenous plants were introduced. In order to limit the intervention by users, setting enough buffer zones, and environment education programs were urgently required. D/H=1>Hyangkyo> houses on the river>temples>lecture halls. D/H ratio of the backside areas is as follows. D/H=1>Hyangkyo>houses on the river>lecture halls. 4. Inner garden were planted deciduous than evergreen trees with Lagerstroemia indica. Enclosed dominant trees were planted by Pinus densiflora, Querces seuata. construct GEM strain, and examined for the expression and functional stability in microcosms.

• Development and Reproduction
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• v.5 no.2
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• pp.151-158
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• 2001

Genomic DNA from the blood of crucian carp(Carassius carassiu) from lake and aquaculture facility in Kunsan, Korea was extracted in order to identify genetic differences by polymerase chain reaction-randomly amplified polymorphic DNAs(PCR-RAPD). Out of 12 primers, 6 generated 266 highly reproducible RAPD markers, producing approximately 2.1 polymorphic bands per primer in crucian carp from lake. The degree of similarity varied from 0.18 to 0.76 as calculated by bandsharing analysis in crucian carp from lake. The RAPD outlines obtained with DNA of two different crucian carp populations from Kunsan were different(0.47 from lake and 0.70 from aquaculture facility, respectively). The electrophoretic analysis of polymerase chain reaction-randomly amplified polymorphic DNAs(PCR-RAPD) products showed high levels of similarity between different individuals in crucian carp from aquaculture facility. This result implies the genetic similarity due to raising in the same environmental condition or inbreeding within the crucian carp from aquaculture facility in Kunsan. In other words, crucian carp may have high levels of genome DNA diversity due to the introduction of the wild population from the other sites of Knsan even if it may be the geographical diverse distribution of this species. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of important aquacultural fish species, crucian carp. However, in future, additional populations and sampling sites will be necessary to complement weak points.

• Research in Plant Disease
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• v.26 no.2
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• pp.79-87
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• 2020

Fusarium wilt caused by Fusarium oxysporum f. sp. fragariae (Fof) is the most important diseases of a strawberry field in Korea. We surveyed phylogenetic analysis, pathogenicity test, and fungicide response about Fof isolates isolated from Korea. Twenty-seven isolates of F. oxysporum isolated from strawberry plants were conducted in this study. Specific amplification by Fof specific primer was confirmed in all 26 isolates except Fo080701 isolate. The nuclear ribosomal intergenic spacer region and the translation elongation factor EF-lα gene sequences of isolates revealed three main lineages. Most of all isolates were contained DNA lineage group 1, but 2 and 3 group was shown only one and three isolates, respectively. All isolates were shown in pathogenicity with cv. Seolhyang. The EC₅₀ mean values of prochloraz ranged 0.02-0.1 ㎍/ml except for Fo080701 and effectively inhibited mycelial growth at low concentrations. The EC₅₀ value of metconazole was also 0.04-0.22 ㎍/ml, showing a similar inhibitory effect to that of prochloraz. The EC₅₀ value of pyraclostrobin was 0.23-168.01 ㎍/ml, which was different according to the strain. In the field trial, boscalid+fludioxonil, fluxapyroxad+pyraclostrobin, and prochloraz manganese were selected as the effective fungicides for controlling Fusarium wilt.

• Restorative Dentistry and Endodontics
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• v.25 no.4
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• pp.587-598
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• 2000

Urea in the oral cavity is hydrolyzed mainly by bacterial ureases to ammonia, which in turn, raises pH of the oral environment, maintaining oral pH homeostasis, thereby inhibiting dental caries. Streptococcus salivarius has been shown to be a major contribution to oral ureolysis. Synthesis of urease by S. salivarius appears to be constitutive, but can be greatly enhanced in the acidic environment. It has been presumed that ureolytic activity of S. salivarius strains isolated from caries-active site is greater than that of strains from caries-free site. However, no in vivo study has supported the presumption. The present study was performed to observe the ureolytic activity of S. salivarius strains isolated from different environments in the same individual, finding out whether the ureolytic activity is related to dental caries. For the purpose, S. salivarius strains were isolated from caries-active site (>C2), a caries-free site of the tooth, and the dorsum of the tongue of each of 50 patients having decayed teeth. The strains isolated from the patients who harbored S. salivarius in more than two sites were selected and then their ureolytic activities were measured. In order to examine clonal diversity of the strains, their ureC genes were amplified by polymerase chain reaction (PCR) and then restricted with EcoRV, and the protein profiles of the strains were compared by SDS-PAGE. The results were as follows: 1. Of 50 patients, 13 patients harbored S. salivarius in more than two sites; a total of 61 S. salivarius strain were isolated from the patients and selected for the study. 2. Of 17 isolates from the caries-active site of 9 patients harboring S. salivarius in more than two sites including carious lesion, 10 (58.8%) showed a high ureolytic activity (> 200 ${\mu}mol/min/mg$). While, 19 out of 44 isolates (43.2%) from the caries-free site of the teeth and the dorsum of the tongues of 13 patients were the strains with a high ureolytic activity. 3. Of 9 patients harboring S. salivarius in more than two sites including caries-active site. 6 patients were found to have the strains in the caries-active site showing a lower ureolytic activity than the strains in the other sites. 4. Of 34 isolates with ureolytic activity higher than 40 ${\mu}mol/min/mg$, 32 isolates produced 0.54-Kbp PCR products regardless of the sites of bacterial collection. In contrast, of 27 isolates with ureolytic activity lower than 40${\mu}mol/min/mg$, 26 isolates yielded 1.3-Kbp PCR products or none regardless of the sites. 5. Different clonal types of S. salivarius with relatively higher and lower ureolytic activities were found in the same individuals and even in the same sites. 6. None of strains showing different ureolytic activity appeared to be the same clonal type. The overall results suggest that ureolytic activity of the isolates does not appear to be related to differences of the environments but related to their own genetic traits.

• Korean Journal of Ecology and Environment
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• v.38 no.2 s.112
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• pp.137-145
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• 2005

Traditional screening techniques have missed up to 99% of microbial resources existing in the nature. Strategies of direct cloning of environmental DNAs comprising tine genetic blueprints of entire microbial metagenomes provide vastly more genetic information than is contained in the culturable. Therefore, one way to screening the useful gene in a variety of environments is the construction of metagenomic DNA library. In this study, the water samples were collected from Daecheong Reservoir in the mid Korea, and analyzed by T-RFLP to examine the diversity of the microbial communities. The crude DNAs were extracted by SDS-based freezing-thawing method and then further purified using an $UltraClean^{TM}kit$ (MoBio, USA). The metagenomic libraries were constructed with the DNAs partially digested with EcoRI, BamHI, and SacII in Escherichia coli DH10B using the pBACe3.6 vector. About 14.0 Mb of metagenomic libraries were obtained with average inserts 13 ${\sim}$ 15 kb in size. The genes responsible for degradation of 1, 2-dichloroethane (1, 2-DCE) via hydrolytic dehalogenation were identified from the metagenomic libraries by colony hybridization. The 1, 2-dichloroethane dehalogenase gene (dhlA) was cloned and its nucleotide sequence was analyzed. The activity of the 1, 2-DCE dehalogenase was highly expressed to the substrate. These results indicated that the dhlA gene identified from the metagenomes derived from Deacheong Reservoir might be useful to develop a potent strain for degradation of 1, 2-DCE.