• Microbiology and Biotechnology Letters
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• v.51 no.4
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• pp.474-483
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• 2023

Members of the genus Streptomyces produce more than 70% of antibiotics. The rise in antibiotic resistance globally enhanced the search for novel species with the ability to produce new bioactive compounds. This study was initiated to investigate different regions in Jordan for previously uncultured and rare Streptomyces species capable of producing novel antimicrobial compounds especially active against bacteria resistant to antibiotics. A total of 191 Streptomyces strains were isolated from 26 soil samples collected from different geographic regions in Jordan. Isolates were characterized based on colony and cellular morphology as well as using 16S rRNA gene sequencing. These isolates were screened for their ability to produce antibiotics by the perpendicular-cross streak method, and then tested by well diffusion method against tested pathogens. Fifty-four isolates showed potential to produce antimicrobial products especially active against resistant bacteria, 20.1% of the isolates showed inhibitory effect against Staphylococcus aureus, 16.9% against clinical MSSA strains, and 18.0% against MRSA: whereas only 4.2% against Esherichia coli, 3.2% against Klebsiella pneumonia, 2.7% against Pseudomonas aeruginosa, and 10.0% against clinical Candida albicans. Three isolates were selected for further identification due to their antibacterial activity against S. aureus, MRSA, and MSSA. These isolates were identified as follows; Streptomyces aburaviensis DSa3, Streptomyces alboniger SAb7 and Streptomyces misionensis ZAb2, based on cultural, biochemical characteristics and molecular analysis of the 16S rRNA.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.104-110
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• 2004

To determine evolution and genotype of new chromosomal AmpC $\beta$-lactamases among clinical isolates of Enterobacter species, we performed antibiotic susceptibility testing, pI determination, sequencing, and phy-logenetic analysis using developed expression/secretion vector. Six isolates have shown to produce AmpC $\beta$-lactamases. Six genes of AmpC $\beta$-lactamases that are responsible for the resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin, and amoxicillin-clavulanic acid were cloned and characterized in pMSG12119. Insert fragment containing the ampC genes was sequenced and found to have an open reading frame coding for 381-amino-acid $\beta$-lactamase. The nucleotide sequence of four ampC genes ($bla_EcloK992004.l$, $bla_EcloK995120.1$, $bla_EcloK99230$, and $bla_EareK9911729$) shared considerable homology with that of chromosomal ampC gene ($bla_EcloMHN1$) of E. cloacae MHN1 (more than 99.6% identity). The sequences of two ampC genes ($bla_EcloK9973$ and $bla_EcloK9914325$) showed close similarity to the chromosomal ampC gene ($bla_EcloQ908R$) of E. clo-acae 908R (99.7% identity). The results from phylogenetic analysis suggested that six ampC genes could be originated from $bla_EcloMHN1$ / or $bla_EcloQ908R$ / MIC patterns and exact pI values of six transformants indicated that the developed expression/secretion vector (pMSG1219) was suitable for the characterization of foreign genes in E. coli strain.

• Korean Journal of Soil Science and Fertilizer
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• v.25 no.4
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• pp.387-393
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• 1992

The fate of inoculum strain of Bradyrhizobium japonicum was studied by using genetically marked strain. RJB6 $str^rnal^rneo^r$. A spontaneous mutant of B. japonicum isolated from nodules was made to have antibiotic resistance against streptomycin and nalidixic acid. In order to make genetically marked strain, neomycine resistant gene(Tn5) was introduced into this spontaneous mutant by conjugation with E. coli containing pSUP2021. The southern hybridization was carried out to confirm the plasmid insertion. Hybridization of chromosome DNA using pSUP2021(Tn5) as a probe showed that Tn5 was located on the 4.9kb fragment of chromosome. Soybean seeds were planted into a soil previously cultivated with soybean and inoculated with different cell densities of marked strain. Fourty days after planting, the inoculation effects on nodule number, nodule fresh weight, plant height and nitrogen content in the plot inoculated with heavy cell suspension was a little better than those in the plot with low inoculation. The recovery percentage of the marked strains was about 12% in the plot inoculated with heavy density cell suspension, while 5% in the plot inoculated with low cell suspension.

• KSBB Journal
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• v.14 no.4
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• pp.460-464
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• 1999

Methicillin-resistant Staphyloccus aureus (MRSA) has been known to be resistant to many kinds of antibiotics and causes a problem of nosnocomial infection since the third generation of cephalosporines has been introduced in the 1980s. As antibiotic sensitivity tests which have been routinely used to detect MRSA in the laboratory depend on the culture conditions such as, pH, temperature, and time, etc., it is difficult to decide in the case of borderline- or low-level of MRSA. Therefore it would be necessary to develope a new method based on the molecular biological technique to overcome these problems. In this study, we extracted DNA from S. aureus and performed polymerase chain reaction (PCR) to amplify mec A gene, encoding penicillin-binding protein 2' (PBP-2'), which is known to confer bacteria resistance to the bacteriostatic action of methicillin. The results were compares with those of minimal inhibitory concentration (MIC) test. When MIC test with oxacillin was performed on the 120 isolates of S. aureus from each patient's specimens, 64 of them were MRSA and 56 of them were methicillin-sensitive Staphylococcus aureus (MSSA). In pus specimen, more precisely, 61.9% (26/42) of MRSA was detected, and 44.2% (19/43), 60% (9/15) and 50% (10/20) of MRSA were detected in sputum, body fluid, and other specimen respectively. When 40 isolates of MRSA and MSSA were tested by PCR method and compares with the results of MIC method, different results were obtained from 1 isolate of MRSA (2.5%) and in 2 isolates of MSSA (5%) suggesting that PCR method should be performed at the same time for more accurate clinical test of MRSA.

• Pediatric Infection and Vaccine
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• v.26 no.3
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• pp.148-160
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• 2019

Purpose: This study aimed to investigate the molecular epidemiology of a methicillin-resistant Staphylococcus aureus (MRSA) outbreak at a newborn nursery and neonatal intensive care unit (NICU). Methods: During the outbreak, from August to September 2017, MRSA isolates collected from neonates and medical staff underwent genotyping and screened for virulence factors. Antibiotic susceptibilities were tested. Results: During the study period, 41 neonates were admitted at the nursery (n=27) and NICU (n=14). Of these, 7 had MRSA infections (skin infection [n=6] and sepsis [n=1]) and 4 were colonized with MRSA. Associated medical staff (n=32) were screened; three were nasal MRSA carriers. Staphylococcal chromosomal cassette mec (SCCmec) type II, sequence type (ST) 89, spa type t375 was found to be the skin infection outbreak causing strain, with multi-drug resistance including low-level mupirocin resistance. SCCmec type IVa, ST 72, and a novel spa type designated t17879, was the cause of MRSA sepsis. Many different types of MRSA were colonized on the neonates; however, SCCmec type IVa, ST 72, spa type t664 was colonized in both neonates and a NICU nurse. All MRSA isolates from colonized infants were positive for the Panton-Valentine leukocidin (PVL) toxin gene. Conclusions: The strain causing an outbreak of skin infections had multi-drug resistance. Also, MRSA colonized in the neonates were found to carry the PVL toxin gene. Because different strains are present during an outbreak, molecular epidemiologic studies are important to identify the outbreak strain and colonized strains which aid in effective control and prevention of future MRSA outbreaks.

• Korean Journal of Microbiology
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• v.55 no.3
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• pp.274-277
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• 2019

Ruminococcus sp. KGMB03662 was isolated from fecal samples obtained from a healthy Korean. The whole-genome sequence of Ruminococcus sp. KGMB03662 was analyzed using the PacBio Sequel platform. The genome comprises a 2,707,502 bp chromosome with a G + C content of 43.09%, 2,484 total genes, 2,367 protein-coding gene, 14 rRNA genes, and 53 tRNA genes. In the draft genome, genes involved in the hydrolysis enzyme, fatty acid biosynthesis, fatty acid metabolite, antibiotic biosynthesis, and antibiotic resistance have been identified. Those genes of KGMB03662 may be related to the regulation of human health and disease.

• Korean Journal of Food Science and Technology
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• v.37 no.1
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• pp.134-141
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• 2005

To characterize genotypic and phenotypic traits of Staphylococcus aureus isolates (n = 86) from lettuces and raw milk, major virulence-associated genes and antibiotic susceptibility were detected using PCR-based methods and disk diffusion method, respectively. All isolates possessed coagulase gene and showed five polymorphism types [500 bp (2.4%), 580 bp (17.4%), 660 bp (61.6%), 740 bp (17.4%), and 820 bp (1.2%)] due to variable numbers of tandem repeats present within the gene. Two or three different loci of hemolysin gene family were dominant in isolates, 47 of which (55%) possessed combination of hla/hld/hlg-2 genes as the most prevalent types. Among enterotoxin-encoding genes, sea was detected from 32 isolates (37%), sed from 1 isolate (1%), and sea and sed genes were co-detected from 4 isolates (5%), whereas seb, sec, and tsst-1 genes were not detected. All isolates were susceptible to ciprofloxacin, trimethoprim/sulfamethoxazole, oxacillin, and vancomycin, 85 isolates (99%) to penicillin G, 54 isolates (63%) to chloramphenicol, 51 isolates (59%) to erythromycin, and 7 isolates (8%) to clindamycin. Among resistant isolates, seven displayed multiantibiotic-resistance against two different antibiotics.

• Korean Journal of Microbiology
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• v.43 no.4
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• pp.256-263
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• 2007

We performed the biochemical characteristics, molecular epidemiologocal analysis, and drug susceptibility test on V. vulnificus isolated from environmental sources in Incheon. For this study, 233 strains were isolated from seawater, sediment, shellfish. V. vulnificus isolates were divided into 15 biochemical groups, which were positive for ONPG and Amygdalin test. Among the 209 strains, 206 (98.6%) strains and 110 (52.6%) strains revealed positive for vvhA and viuB gene, and the viuB gene detection rates of V. vulnificus from seawater, shellfish and sediment were 48%, 48.5% and 61.6%, respectively. From disc diffusion test on 175 isolates, most of strains were sensitive to Imipenem (100.0%), Sulfamethoxazole/trimethoprim (98.9%), Tetracycline, Ciprofloxacin (98.3%), Ampicillin/sulbactam (97.1%), Ohloramphenicol (96.6%), Cefepime (94.9%) and Ceftriaxone (94.8%), multi-drug resistance rates was 31.5% of seawater, 34.4% of sediment and 29.2% of shellfish. PFGE was performed on 233 V. vulnificus isolates with the objective of investigating the extent of genetic diversity of these isolates in our region. We could find that at least 126 different PFGE patterns were generated according by 90% of similarity and 13 clusters by 58% of similarity. The major cluster was type I (44.6%) during the most of the year, and type J was frequent pattern in June and October. There were 9 distinct PFGE types in July, 8 types in August, 7 types in June, 6 types in September, 5 types in October 3 types in May and 1 type in March.

• Korean Journal of Food Science and Technology
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• v.37 no.3
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• pp.456-464
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• 2005

Antimicrobial drug-resistance is natural response to antimicrobial stress based on selection, which weakens chemotherapy effect. Introduction of large numbers of chemotherapeutic agents to clinical practice has generated strains of microorganisms that survive and multiply in vivo with high-drug concentrations. Methicillin-resistant Staphylococcus aureus (MRSA), bacteria found in normal daily life, can be easily ingested through milk vegetables, and meats, etc. MRSA emerged in many port of the world, increasing complex clinical problems. Therefore, new agents are needed to treat MRSA. Glycyrrhiza uralensis was extracted using 80% MeOH to investigate its antimicrobial activity against MRSA stains KCCM 11812, 40510, and 40512 through bacterial measurement, disc diffusion, and O.D. methods, MIC values, MRSA gene expression investigation, and scanning electron microscope observation. Results revealed MecA, Mecl, MecRI, and FemA were the most highly manifested MRSA genes. Methanolic extract of G. uralensis significantly inhibited MRSA and thus could be used in development of antibacteria.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.305-310
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• 2008

Staphylococcus aureus is one of the most significant pathogens and a causative agents of nosocomial infections. The emergence of methicillin resistant S. aureus (MRSA), in particular, has become a major clinical and epidemiological problems worldwide. In this study, we analyzed the toxin genes and investigated molecular epidemiological characteristics of S. aureus isolated from stools of diarrheal patients at the hospitals in Incheon. Of the 609 strains from 2,281 specimens, 173 strains retained enterotoxin; 68 isolates (39.30%), 100 isolates (57.80%) were classified to A and C type, respectively. In the antibiotic susceptibility, all of enterotoxin positive isolates were resistant to oxacillin. Eighty eight strains (50.86%) of 173 MRSA isolate possessed tsst gene, but eta and eth genes were not detected at all. In the detection of MRSA associated genes by PCR method, mecA genes were detected in 167 strains (96.53%). From the result of PFGE analysis, we classified tsst-positive MRSA to 10 types and 24 subtypes. Type A, H and F were the major strains comprised of 57.95% (51 strains), 10.22% (9 strains) and 9.09% (8 strains) respectively.