• Journal of Life Science
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• v.28 no.11
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• pp.1301-1315
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• 2018

We purified an antimicrobial peptide from the acidified hemocyte extract of Mytilus coruscus by $C_{18}$ reversed-phase high-performance liquid chromatography (RP-HPLC). The peptide was 4041.866 Da based on matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS) and the 25 amino acids of the N-terminus sequence were identified. Comparison of this sequence of the purified peptide with the N-terminus sequences of other antimicrobial peptides revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We also identified a 312 bp open-reading frame (ORF) encoding 103 amino acids based on the obtained amino acid residues. The nucleotide sequence of this ORF and the amino acid sequence also revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We synthesized two antimicrobial peptides with an alanine residue in the C-terminus, and designated them mytilin B1 and B2. These two antimicrobial peptides showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus and Streptococcus parauberis (minimal effective concentration, MECs $41.6-89.7{\mu}g/ml$), gram-negative bacteria, including Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Providencia stuartii, Pseudomonas aeruginosa, and Vibrio ichthyoenteri (MECs $7.4-39.5{\mu}g/ml$), and the fungus Candida albicans (MECs $26.0-31.8{\mu}g/ml$). This antimicrobial activity was stable under heat and salt conditions. Furthermore, the peptides did not exhibit significant hemolytic activity or cytotoxic effects. These results suggest that mytilin B could be applied as alternative antibiotic agent, and they add to the understanding of the innate immunity of hard-shelled mussels.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.410-419
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• 2018

It is urgently required to construct safety data on agricultural by-products imported for use as medium materials for domestic mushroom production. However, research on microorganisms is insufficient. This study was conducted to investigate the presence of bacteria that have the possibility of harmful effects on human, plants and mushroom in wheat straw, peatmoss, cottonseed hull, cottonseed meal, and beet pulp imported from Australia, Canada, China, Egypt, Germany. Bacteria were found in the range of $1.35{\times}10^2$ to $8.34{\times}10^6CFU/g$. As a result of 16S rDNA sequence analysis, total of 19 genera and 45 species of bacteria were identified. Bacillus genus was dominant, followed by Paenibacillus genus. At the species level, diverse species was in the order of Firmicute, Proteobacteria and Actinobacteria. Regarding the agricultural by-products, straw and peat moss had more diverse bacteria than other agricultural by-products. Among the indentified bacteria, 6 species of 5 genera (Enterobacter asburiae, Enterobacter ludwigii, Stenotrophomonas maltophilia, Pseudomonas monteilii, Bacillus anthracis, and Cellulosimicrobium funkei) were present as potent harmful bacteria to human. Surprisingly, both the human and plant pathogenic Klebsiella pneumoniae subsp. pneumonia was present. Bacillus altitudinis was present as a plant pathogen. Lysinibacillus sphaericus, an insect pathogen, and Ochrobactrum pseudogrignonense, a mushroom pathogen, were also present. The results of this study confirmed that several kinds of pathogenic bacteria were present in the agricultural by-products for the mushroom cultivation medium imported into Korea. Our work suggests that hygiene inspection and management is urgently needed for imported agricultural by-products to be safely used for mushroom production.

• Annals of Clinical Microbiology
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• v.22 no.1
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• pp.9-13
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• 2019

Background: The isolation of carbapenemase-producing Enterobacteriaceae (CPE) has become increasingly common. Continuous surveillance for these organisms is essential because their infections are closely related to outbreaks of illness and are associated with high mortality rates. The aim of this study was to develop and evaluate multiplex PCR as a means of detecting several important CPE genes simultaneously. Methods: We aimed to develop a multiplex PCR that could detect seven CPE genes simultaneously. The multiplex PCR was composed of seven primer sets for the detection of KPC, IMP, VIM, NDM-1, GES, OXA-23, and OXA-48. We designed different PCR product sizes of at least 100 bp. We evaluated the performance of this new test using 69 CPE-positive clinical isolates. Also, we confirmed the specificity to rule out false-positive reactions by using 71 carbapenem-susceptible clinical strains. Results: A total of 69 CPE clinical isolates showed positive results and were correctly identified as KPC (N=14), IMP (N=13), OXA-23 (N=12), OXA-48 (N=11), VIM (N=9), GES (N=5), and NDM (N=5) by the multiplex PCR. All 71 carbapenem-susceptible clinical isolates, including Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa, showed negative results. Conclusion: This multiplex PCR can detect seven CPE genes at a time and will be useful in clinical laboratories.

• CELLMED
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• v.4 no.1
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• pp.5.1-5.8
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• 2014

Healthcare-associated infection represents a frequent cause of mortality that increases hospital costs. Due to increasing microbial resistance to antibiotics, it is necessary to search for alternative therapies. Consequently, novel alternatives for the control of resistant microorganisms have been studied. Among them, plant antimicrobial protein presents enormous potential, with flowers being a new source of antimicrobial molecules. In this work, the antimicrobial activity of protein-rich fractions from flower tissues from 18 different species was evaluated against several human pathogenic bacteria. The results showed that protein-rich fractions of 12 species were able to control bacterial development. Due its broad inhibition spectrum and high antibacterial activity, the protein-rich fraction of Hibiscus rosa-sinensis was subjected to DEAE-Sepharose chromatography, yielding a retained fraction and a non-retained fraction. The retained fraction inhibits 29.5% of Klebsiella pneumoniae growth, and the non-retained fraction showed 31.5% of growth inhibition against the same bacteria. The protein profile of the chromatography fractions was analyzed by using SDS-PAGE, revealing the presence of two major protein bands in the retained fraction, of 20 and 15 kDa. The results indicate that medicinal plants have the biotechnological potential to increase knowledge about antimicrobial protein structure and action mechanisms, assisting in the rational design of antimicrobial compounds for the development of new antibiotic drugs.

• Korean Journal of Veterinary Service
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• v.28 no.1
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• pp.1-21
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• 2005

Pullorum disease and Fowl typhoid are kind of poultry specific disease for poultry. The peculiar character of these poultry specific diseases is that it can be infected by transmitting vertically and horizontally, also it is hard to be discovered by clinical sign, and pathology or immunology. So, to develop the PCR method which distinguishes these two genetically similar diseases of separated the specific DNA fragment from each strain and use it for differential diagnosis by subtraction PCR method. Standard strain of S gallinarum and S pullorum, and field isolation strain were verified by biochemistry, It confirmed existence of plasmid by using the PFGE. Then, Isolated DNA from it and used it as materials for the experiment. After cutting genomic DNA of two strains by using Sau 3Al, It ligated primer to tester DNA for PCR amplification and separated specific DNA fragment bacteria with method of subtraction PCR. And, It confirmed that it is a piece of unique DNA in every bacteria using base sequence of separated DNA fragment. 1. The six specific DNA fragment were separated from the DNA of S gallinarum and S pullorum by the subtraction PCR method. 2. In the result of comparison after setting base sequence of each fragment, each separated base sequence of DNA fragment they did not correspond to each other 3. As the result of each DNA fragment is derived from the each strain of DNA, and there was no homology of genomic DNA level in mutual. 4. The fragment originated in plasmid and includes S pullorum did not separate. 5. In the result of searching base sequence in Genebank, it partially shows homology in Salmonella enterica, S typhimurium, S dublin, Escherichia coli, Shigella flexneri, Yersinia pestis, Klebsiella pneumoniae. 6. Primer design by S gallinarum DNA 2, 3 fragment used PCR, They are positive reaction in only S gallinarum at 276, 367 bp position.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.5
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• pp.436-446
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• 2006

The purpose of this study was to isolate and identify the bacteria in chronic maxillary sinusitis (CMS) lesions from 3 patients and to determine the antimicrobial susceptibility of them against 10 antibiotics. One of them was odontogenic origin and the others were non-odontogenic origin. Pus samples were collected by needle aspiration from the lesions and examined by culture method. Bacterial culture was performed in three culture systems (anaerobic, CO2, and aerobic incubator). Identification of the bacteria was performed by 16S rRNA gene (16S rDNA) nucleotide sequencing method. To test the sensitivity of the bacteria isolated from the maxillary sinusitis lesions against seven antibiotics, penicillin G, amoxicillin, tetracycline, ciprofloxacin, cefuroxime, erythromycin, clindamycin, and vancomycin, minimum inhibitory concentration (MIC) was performed using broth dilution assay. Our data showed that enterobacteria such as Enterobacter aerogenes (30%), Klebsiella pneumoniae (25%), and Serratia marcescens (15%) were predominately isolated from the lesion of non-odontogenic CMS of senile patient (70 year old). Streptococcus spp. (40.3%), Actinomyces spp. (27.4%), P. nigrescens, M. micros, and P. anaerobius strains were isolated in the lesion of odontogenic CMS. In the lesion of non-odontogenic CMS, Streptococcus spp. (68.4%), Rothia spp. (13.2%), and Actinomyces sp. (10.5%) were isolated. The susceptibility pattern of 10 antibiotics was determined according to the host of the bacteria strains ratter than the kinds of bacterial species. Even though the number of CMS was limited as three, these results indicate that antibiotic susceptibility test must be accompanied with treatment of CMS. The combined treatment of two or more antibiotics is better than single antibiotic treatment in the presence of multidrug-resistant bacteria in the CMS lesions.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1085-1091
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• 2009

A phytase with high activity at low temperatures has great potential for feed applications, especially in aquaculture. Therefore, this study used a degenerate PCR and TAIL PCR to clone a phytase gene from Erwinia carotovora var. carotovota, the cause of soft rot of vegetables in the ground or during cold storage. The full-length 2.5-kb fragment included an open reading frame of 1,302 bp and encoded a putative phytase of 45.3 kDa with a 50% amino acid identity to the Klebsiella pneumoniae phytase. The phytase contained the active site RHGXRXP and HD sequence motifs that are typical of histidine acid phosphatases. The enzyme was expressed in Escherichia coli, purified, and displayed the following characteristics: a high catalytic activity at low temperatures (retaining over 24% activity at $5^{\circ}C$) and remarkably thermal lability (losing >96% activity after incubation at $60^{\circ}C$ for 2 min). The optimal phytase activity occurred at pH 5.5 and ${\sim}49^{\circ}C$, and the enzyme activity rapidly decreased above $40^{\circ}C$. When compared with mesophilic counterparts, the phytase not only exhibited a high activity at a low temperature, but also had a low $K_m$ and high $k_{cat}$. These temperature characteristics and kinetic parameters are consistent with low-temperature-active enzymes. To our knowledge, this would appear to be the first report of a low-temperature-active phytase and its heterogeneous expression.

• Journal of Microbiology and Biotechnology
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• v.26 no.10
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• pp.1723-1735
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• 2016

Pyrosequencing analysis of intestinal microflora from healthy Thai vegetarians and non-vegetarians exhibited 893 OTUs covering 189 species. The strong species indicators of vegetarians and non-vegetarians were Prevotella copri and Bacteroides vulgatus as well as bacteria close to Escherichia hermanii with % relative abundance of 16.9 and 4.5-4.7, respectively. Core gut microbiota of the vegetarian and non-vegetarian groups consisted of 11 and 20 different bacterial species, respectively, belonging to Actinobacteria, Firmicutes, and Proteobacteria commonly found in both groups. Two species, Faecalibacterium prausnitzii and Gemmiger formicilis, had a prevalence of 100% in both groups. Three species, Clostridium nexile, Eubacterium eligens, and P. copri, showed up in most vegetarians, whereas more diversity of Collinsella aerofaciens, Ruminococcus torques, various species of Bacteroides, Parabacteroides, Escherichia, and different species of Clostridium and Eubacterium were found in most non-vegetarians. Considering the correlation of personal characters, consumption behavior, and microbial groups, the age of non-vegetarians showed a strong positive correlation coefficient of 0.54 (p = 0.001) to Bacteroides uniformis but exhibited a moderate one to Alistipes finegoldii and B. vulgatus. Only a positive moderate correlation of body mass index and Parabacteroides distasonis appeared. Based on the significant abundance of potential pathogens, the microbiota of the non-vegetarian group showed an abundance of potential pathogen varieties of Bilophila wadsworthia, Escherichia coli, and E. hermannii, whereas that of the vegetarian group served for only Klebsiella pneumoniae. These results implied that the microbiota of vegetarians with high abundance of P. copri and low potential pathogen variety would be a way to maintain good health in Thais.

• Journal of Yeungnam Medical Science
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• v.28 no.2
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• pp.133-144
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• 2011

Background: Tigecycline (TIG), a new broad-spectrum glycylcycline with anti-multidrug-resistant-(MDR)-pathogen activity, was launched in March 2009 in South Korea, but there are insufficient clinical studies on its use in the country. As such, this study was performed to analyze cases of severe MDR-pathogen-caused infections treated with TIG. Methods: Patients treated with TIG within the period from May 2009 to June 2010 were enrolled in this study. Their clinical and microbiologic data were reviewed retrospectively. Results: Twenty-one patients were treated with TIG for complicated skin and soft-tissue infections (cSSTIs) (42.9%), complicated intra-abdominal infections (cIAIs) (38.1%), or pneumonia (19.1%) caused by MDR pathogens like carbapenem-resistant $Acinetobacter$ $baumannii$ (76.2%), methicillin-resistant $Staphylococcus$ $aureus$ (61.9%), extended-spectrum beta-lactamase-producing $Escherichia$ $coli$ and $Klebsiella$ $pneumoniae$ (38.1%), and penicillin-resistant $Enterococcus$ species (33.3%). Thirteen patients (61.9%) had successful clinical outcomes while five (23.8%) died within 30 days. The rate of clinical success was highest in cSSTI (77.8%), followed by cIAI (50%) and pneumonia (50%), and the mortality rate was highest in pneumonia (50%), followed by cIAI (25%) and cSSTI (11.1%), Conclusion: Tigecycline therapy can be an option for the treatment of severe MDR-pathogen-caused infections in South Korea, Due to its high risk of failure and mortality, however, prudence is required in its clinical use for the treatment of severe infections like nosocomial pneumonia.

• The Korean Journal of Community Living Science
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• v.17 no.4
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• pp.39-48
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• 2006

This study was carried out to make a search for the usefulness of marigold plants (Tagetes erecta) as a natural dye. And we have also examined into the dyeability and antifungal activity of silk and cotton fabrics dyed with 2% powdered colorant and 1% liquid colorant. Almost surface colors of silk and cotton fabrics dyed with 2% powdered colorant and that dyed with 1% liquid colorant except for that treated Cu and Fe with mordants were shown by Y-level. $L^*$ values of silk fabrics dyed with 2% powdered colorant were 66.2 to 86.39, those $a^*$ values were -1.97 to 6.09, and those $b^*$ values were 13.69 to 35.97. And $L^*$ values of cotton fabrics dyed with 2% powdered colorant were 78.97 to 89.62, those a values were -3.39 to 0.38, and those $b^*$ values were 5.63 to 15.61 $L^*$ values of silk fabrics dyed with 1% liquid colorant were 34.37 to 85.57, those a values were -7.79 to 12.33, and those $b^*$ values were 15.13 to 82.91, And $L^*$ values of cotton fabrics dyed with 1% liquid colorant were 44.12 to 87.90, those $a^*$ values were -8.37 to 6.18, and those $b^*$ values were 11.65 to 78.87. Colorfastness of silk fabrics against light treated nothing with mordants and that dyed with 1% liquid colorant decreased for a little by second grade, however, that against washing, rubbing, perspiration and dry cleaning were increased over fourth grade. Colorfastness of cotton fabrics in terms of rubbing, light, and dry cleaning, except for washing and perspiration, were shown by over third grade, when that was treated nothing with mordants. Antifungal activities of silk fabrics treated nothing with mordants and dyed with 1% liquid colorant of marigold extracts were shown by 28.9% against Staphylococcus aureus. However, those of Al, Ca, Cu and tartaric acid mordanting were more than 25.5% against Staphylococcus aureus and Klebsiella pneumoniae.