• Restorative Dentistry and Endodontics
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• v.20 no.2
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• pp.577-609
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• 1995

Bacteria have been regarded as one of the most important factors in pulpal and periapical diseases. Streptococci are frequently isolated facultative anaerobes in infected root canals. Recently molecular biological techniques have been rapidly progressed. This study was designed to apply the molecular biological tools to the identification and classification of streptococci in the endodontic microbiology. Streptococci isolated from infected root canals were identified with both Vitek Systems and API 20 STREP. Identification results were somewhat different in several strains of streptococci. Eighteen streptococci and enterococcal was difficult so to digest plasmid DNA using Hind III and EcoRI to differentiate strains by restriction enzyme analysis of plasmid DNA. 16S rDNA of chromosome was amplified by polymerase chain reaction(PCR) and then restricition fragment length polymorphism(RFLP) using several restriction enzymes was observed. The molecular mass of 16S rDNA of chromosomal DNA was approximately 1.4kb. There were three to five RFLP patterns using eight restriction enzymes. RFLP patterns digested with CfoI which recognizes four base sequences were identical in all stains. Hind III which recognizes six base sequences could not digest the 16S rDNA. Restriction enzymes which recognize five base sequences were suitable for RFLP pattern analysis. At least three different restriction enzymes were needed to compare each strains. 16S rDNA PCR-RFLP was simple and rapid to differentiate and classify strains and could be used in the epidemiological study of root canal infections.

• Journal of the Korean Wood Science and Technology
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• v.25 no.3
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• pp.50-57
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• 1997

The presence of slime in paper mills is practically universal. Many researches have been performed for many years to resolve the problem caused by the slime in pulp and paper mill. Many papers have been published to show the bacteria is a major cause of paper mill slime. Now that the recycling of the water has been increased and the regulations of a toxic chemical dosage have become more strengthen, the importance of the control of slime in pulp and paper mill recently has been more recognized. Therefore, to produce quality products at the lowest economic and environmental costs, a through study of the microbial ecology and the indentification of troublesome slime-forming bacteria is a quite necessary. The purpose of this paper is to indentify slime~forming bacteria isolated from the papermaking process. The samples were taken from four parts of making fine paper : machine chest, head box, wire part, white water tank. Machine chest showed the most numbers of bacteria, numbering $2.55{\times}10^7$. The different colony types were taken from the 105 dilution plate. Nine bacteria were identified u sing the Biolog system and the vitek system: 6 gram-negative bacteria, 3 gram-positive bacteria. They are Pseudomonas paucimobilis B., Staphylococcus sp., Acinetobacter calcoaceticus., Pseudomonas cepacia, Actinobaci1lus capsulatus, Acidovorax sp., Flavobacterium sp., and Staphylococcus auricularis in addition to one unidentified sp., Among them. Pseudomonas paucimobillis was found in all places where the samples were taken. And, each parts had the different predominant bacteria in it : Pseudomonas paucimobilis B. in machine chest, Acinetobactor calcoaceticus. in Wire Part and Staphylococcus sp. in head box.

• Korean Journal of Veterinary Research
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• v.56 no.1
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• pp.45-46
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• 2016

For several days, there was a series of mortalities of chub mackerel (Scomber japonicus) that were reared for public exhibition in a private aquarium in Seoul, Korea. As part of the diagnosis of the dead fish, a bacterial isolate from the kidney was cultured, identified, and confirmed to be Vibrio (V.) harveyi using Vitek System 2 and 16S rRNA gene sequencing. Phylogenetic analysis was also performed by the neighbor-joining method. As a result, the V. harveyi isolated from chub mackerels of a private aquarium in Korea, called as SNUVh-LW1, was clustered in the same group with V. harveyi ATCC33843.

• Biomedical Science Letters
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• v.25 no.1
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• pp.40-53
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• 2019

Of the Acinetobacter spp., A. baumannii (genospecies 2) is the most clinically significant in terms of hospital-acquired infections worldwide. It is difficult to perform Acinetobacter-related taxonomy using phenotypic characteristics and routine laboratory methods owing to clusters of closely related species. The ability to accurately identify Acinetobacter spp. is clinically important because antimicrobial susceptibility and clinical relevance differs significantly among the different genospecies. Based on the medical importance of pathogenic Acinetobacter spp., the distribution and characterization of Acinetobacter spp. isolates from 123 clinical samples was determined in the current study using four typically applied bacterial identification methods; partial rpoB gene sequencing, amplified rRNA gene restriction analysis (ARDRA) of the intergenic transcribed spacer (ITS) region of the 16~23S rRNA, the $VITEK^{(R)}$ 2 system (an automated microbial identification system) and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). A. baumannii isolates (74.8%, 92/123) were the most common species, A. nosocomialis (10.6%, 13/123) and A. pittii isolates (7.5%, 9/123) were second and third most common strains of the A. calcoaceticus-A. baumannii (ACB) complex, respectively. A. soli (5.0%, 6/123) was the most common species of the non-ACB complex. RpoB gene sequencing and ARDRA of the ITS region were demonstrated to lead to more accurate species identification than the other methods of analysis used in this study. These results suggest that the use of rpoB genotyping and ARDRA of the ITS region is useful for the species-level identification of Acinetobacter isolates.

• Biomedical Science Letters
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• v.25 no.1
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• pp.75-82
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• 2019

We developed the multiplex LAMP assay using 16S rRNA, femA and mecA genes for direct detection of the methicillin resistance in Staphylococci from positive blood culture. To simultaneously recognize Staphylococci genus, S. aureus and methicillin resistance, three sets of six primers for 16S rRNA, femA and mecA were designed, respectively. The performance of LAMP assay was affirmed using VITEK system for the phenotypic methods of identification and for oxacillin and cefoxitin antimicrobial susceptibility. The optimal condition for LAMP assay was obtained under $64^{\circ}C$ for 50 min. The detection limit was determined to be of 20 copies and CFU/reaction ($10^4CFU/mL$). For clinical application of comparison with phenotypic methods, the sensitivity and specificity of the LAMP with femA gene for detecting S. aureus was 95.31% and 100%, respectively. The sensitivity and specificity of the LAMP with mecA gene for detecting methicillin resistance was 98.46% and 100%, respectively. The multiplex LAMP assay with femA and mecA gene successfully detected all of MRSA (38 isolates) isolates from 103 Staphylococci in blood cultures. The LAMP assay developed in this study is sensitive, specific, and of excellent agreement with the phenotypic methods.

• The Plant Pathology Journal
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• v.37 no.2
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• pp.137-151
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• 2021

The present study describes the bacterial blight of walnut, caused by Xanthomonas arboricola pv. juglandis (Xaj) in the northern Gyeongbuk province, Korea. Disease symptoms that appear very similar to anthracnose symptoms were observed in walnut trees in June 2016. Pathogens were isolated from disease infected leaves, fruits, shoots, bud, flower bud of walnut, and cultured onto yeast dextrose carbonate agar plates. Isolated bacteria with bacterial blight symptoms were characterized for their nutrient utilization profiles using Biolog GN2 and Vitek 2. In addition, isolates were subjected to physiological, biochemical, and morphological characterizations. Furthermore, isolates were identified using 16S rDNA sequence analysis, and multi-locus sequence analysis using atpD, dnaK, efp, and rpoD. To confirm pathogenicity, leaves, fruits, and stems of 3-year-old walnut plants were inoculated with bacterial pathogen suspensions as a foliar spray. One week after inoculation, the gray spots on leaves and yellow halos around the spots were developed. Fruits and stems showed browning symptoms. The pathogen Xaj was re-isolated from all symptomatic tissues to fulfill Koch's postulates, while symptoms were not appeared on control plants. On the other hand, the symptoms were very similar to the symptoms of anthracnose caused by Colletotrichum gloeosporioides. When walnut plants were inoculated with combined pathogens of Xaj and C. gloeosporioides, disease symptoms were greater in comparison with when inoculated alone. Xaj population size was more in the month of April than March due to their dormancy in March, and sensitive to antibiotics such as oxytetracycline and streptomycin, while resistant to copper sulfate.

• Journal of Veterinary Science
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• v.21 no.4
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• pp.54.1-54.11
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• 2020

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of severe infections in humans and animals worldwide. Studies elucidating the population structure, staphylococcal cassette chromosome mec types, resistance phenotypes, and virulence gene profiles of animal-associated MRSA are needed to understand spread and transmission. Objectives: The objective of this study was to determine 1) clonal complexes and spa types, 2) resistance phenotypes, and 3) virulence/resistance gene profiles of MRSA isolated from animals in Switzerland. Methods: We analyzed 31 presumptive MRSA isolates collected from clinical infections in horses, dogs, cattle, sheep, and pigs, which had tested positive in the Staphaurex Latex Agglutination Test. The isolates were characterized by spa typing and DNA microarray profiling. In addition, we performed antimicrobial susceptibility testing using the VITEK 2 Compact system. Results: Characterization of the 31 presumptive MRSA isolates revealed 3 methicillinresistant Staphylococcus pseudintermedius isolates, which were able to grow on MRSA2 Brilliance agar. Of the 28 MRSA isolates, the majority was assigned to CC398 (86%), but CC8 (11%) and CC1 (4%) were also detected. The predominant spa type was t011 (n = 23), followed by t009 (n = 2), t034 (n = 1), t008 (n = 1), and t127 (n = 1). Conclusions: The results of this study extend the current body of knowledge on the population structure, resistance phenotypes, and virulence and resistance gene profiles of MRSA from livestock and companion animals.

• Journal of Environmental Health Sciences
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• v.50 no.4
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• pp.274-281
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• 2024

Background: One of the major causes of pathogenic E. coli is the feces of infected livestock, and the management of the livestock environment is necessary to prevent pathogenic E. coli. Objectives: The prevalence of pathogenic E. coli was identified from livestock environments, and the molecular characteristics and antibiotic resistance profiles of the isolated pathogenic E. coli strains were analyzed. Methods: In 2022 and 2023, nine points of livestock houses at sites in Chungcheongnam-do Province were selected, and 100 cow feces or soil samples around the livestock houses were collected once per month. Pathogenic E. coli was isolated by selective culture and identified using multiplex PCR. Antibiotic resistance was tested on the isolated strains by using VITEK-2, and candidate strains were selected to perform 16s rRNA sequencing and phylogenetic analysis. Results: A total of 100 samples were tested, and 60 pathogenic E. coli strains were isolated. Of these, 45 and 15 isolates were determined to be single and hybrid pathogenic E. coli , respectively. Among the 15 hybrid pathogenic E. coli strains, eight, five, and two strains were respectively identified as EHEC/ETEC, EHEC/EPEC, and EHEC/ETEC/EPEC hybrids. All 45 isolates showed resistance to at least one antibiotic, and they were susceptible to cefotaxime, amikacin, nalidixic acid, and ciprofloxacin. The highest resistance was against cefalotin, tetracyclin, and ampicillin (20.0%~58.3%). The 16s rRNA sequences of candidate isolates revealed nucleotide sequence identities of 99.1% to 100%. Conclusions: In order to manage pathogenic E. coli from the One Health animal environment perspective, the characteristics of the occurrence of pathogenic E. coli from the livestock environment and molecular biology and antibiotic resistance to isolated strains were analyzed. In order to prevent and manage the occurrence of pathogenic E. coli, these monitoring studies must be continuously conducted.

• Childhood Kidney Diseases
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• v.8 no.2
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• pp.214-222
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• 2004

Purpose: Appropriate antibiotic therapy is important in childhood urinary tract infection and the selection of anibiotics is based on antimicrobial sensitivity of Escherichia coli. Extended-Spectrum ${\beta}-Lactamase(ESBL)$ is an enzyme produced by gram-negative bacilli that has the ability to hydrolyse penicillins, broad-spectrum cephalosporin and monobactam. There have been many reports of outbreaks of hospital infection by ESBL-producing organism. However, community-acquired infection with ESBL-producing organism are rare. This study was performed to retrospectively identify the incidence, characteristics and risk factors of ESBL (+) E. coli in community-acquired childhood UTI. Methods: In 288 children admitted in Ewha Womans University Hospital with E. coli UTI from Mar 2001 to February 2003, ESBL was isolated. ESBL was confirmed by the utilization of an automatized machine(Vitek GNS 433 card) using liquid medium dilution method according to National Committee for Clinical Laboratory Standard. The clinical characteristics, risk factors, antimicrobial resistance and treatment effectiveness were compared with ESBL(-) E. coli UTI. Results: Of 288 E. coli isolates, 31(10.8%) produced ESBL and 93.5%(29/31) occurred in infants younger than 6 month of age(P<0.01). No significant differences were noted in prior antibiotic use, prior admission history and underlying urogenital anomaly. Antimicrobial resistance was significantly higher in ESBL(+) E. coli compared with control patients (P<0.05). Although ceftriaxone showed 100% resistance in ESBL(+) E. coli, bacteriologic sterilization rate after ceftriaxone therapy was higher(96.8%). However, the recurrence rate of febrile UTI within 6 months was higher(25.8%) than control patients(6.6%). Conclusion: Epidemiologic study is required to find out any new risk factors of community-acquired ESBL(+) E. coli UTI and changes in selection of empirical antibiotics should be considered.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.342-351
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• 2006

The aim of this study was to survey susceptibilities of Escherichia coli and Klebsiella pneumoniae isolates against cefotaxime and to determine the prevalences of CTX-M type extended-spectrum $\beta$-lactamases (ESBLs) producing E. coli and K. pneumoniae in Korea. During the period of February to July, 2005, 153 E. coli and 52 K. neumoniae isolates were collected from 2 hospitals in Busan. Antimicrobial susceptibilities to cefotaxime were tested by the disk diffusion method. ESBL production of E. coli and K. pneumoniae was determined by the double disk synergy test. MICs of $\beta$-lactam antibiotics were determined by the agar dilution method. Blac$_{CTX-M}$ genes of the organism were detected by PCR. Among 153 isolates of E. coli and 52 isolates of K. neumoniae, 27 (17.6%) and 25 (48.0%) were intermediate or resistant to cefotaxime, respectively. Twenty-three (15.0%) isolates out of 153 E. coli and 13 (25.0%) out of 52 K. neumoniae isolates showed positive results for ESBL by the double disk synergy test. Twenty isolates out of 23 ESBL producing E. coli and 12 out of 13 ESBL producing K. neumoniae isolates harbored biacTx-M gene,11 of ESBL producing E. coli and 12 of ESBL producing K. neuinoniae isolates harbored bla$_{CTX-M}$ gene, 11 of the ESBL producing E. coli and 2 of ESBL producing K. neumoniae isolates harbored bla$_{TEM}$ gene, and 1 of the ESBL producing E. coli and 12 of ESBL producing K. neumoniae isolates harbored bla$_{SHV}$ gene. E. coli and K. neumoniae isolates producing CTX-M-type ESBLs were not uncommon in Korea. It is thought that continuous survey are necessary for inspecting the spread and novel variants of CTX-M-type ESBL genes. Further me]'e investigation and research on ESBL producing strains are needed in order to prevent the spread of resistant bacteria.