• Korean Journal of Microbiology
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• v.37 no.1
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• pp.72-79
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• 2001

The effect of phenol on the change of bacterial community in the effluent water from a wastewater treatment plant was analyzed by PCR and terminal restriction fragment length polymorphism (T-RFLP). The fragments of 16S rDNA were amplified by PCR with bacterial primers, where one of the primers was biotinylated at the 5'-end. After digestion with restriction enzymes, HaeIII and AluI, the biotinylated terminal restriction tragments (T-RFs) of the digested products were selectively isolated by using streptavidin paramagnetic particles. The single-stranded DNA of T-RFs was separated by electrophoresis on a polyacrylamide gel and detected by silver staining technique. When 10 standard strains were analyzed by our method, each strain had a unique T-RF which corresponded to the calculated size from the known sequences of RDP database. The T-RFLP fingerprint generated from the effluent water was very complex, and the predominant T-RFs corresponded to members of the genus Acinetobacter, Bacillus and Pseudomonas. In addition, the perturbation of bacterial community was observed when phenol was added to the sample at the final concentration of 250 $l^{-1}$. The number of T-RFs increased and the major bacterial population could be assigned to the genus Acinetobacter, Comamonas, Cytophaga and Pseudomonas. A intense band assigned to the putative genera of Acinetobacter and Cytophaga was eluted, amplified, and sequenced. The nucleotide sequence of the T-RF showed close relationship with the sequence of Acinetobacter junii.

• Korean Journal of Microbiology
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• v.50 no.4
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• pp.351-359
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• 2014

A beverage was produced by the fermentation of mixed extracts from the various fruits, vegetables, algae, and medical herbs. The physicochemical properties of the fermented beverage of plant extracts (FBPE) and microbial diversity were analyzed in cultures enriched from FBPE using 16S and 26S rRNA gene sequence analyses. The pH, acidity, $^{\circ}brix$, reducing sugar, and alcohol contents of the FBPE were determined to be the 3.48, 1.68%, 70.0, 1,026 g/L, and 3.5%, respectively. The most abundant free sugar and organic acid in the FBPE were glucose (567.83 g/L) and tartaric acid (93.68 mg/L), respectively. Lactobacillus homohiochii was the predominant species in all enriched culture samples: 100% of the species in 0B (0% sugar) and 40B (40% sugar) libraries and 95.6% of 20B library (20% sugar). Lactobacillus fructivorans was detected in the 20B library. The predominant species in the samples of enrichment cultures collected from FBPE with three different sugar concentrations were: Candida zeylanoides (45.2%) in the 0Y library (0% sugar), Candida lactis-condensi (35.7%) and C. zeylanoides (35.7%) in the 20Y library (20% sugar), and C. lactis-condensi (38.1%) in the 40Y library (40% sugar). This result may provide a useful frame of reference for further analyses of microbial population dynamics in FBPE.

• Research in Plant Disease
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• v.16 no.1
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• pp.94-96
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• 2010

In May of 2004 through 2007, Leptosphaerulina leaf blight caused by Leptosphaerulina trifolii occurred on Kentucky bluegrass (Poa pratensis) at golf courses in Gangwon Province, Korea. Symptoms on the turfgrass caused by L. trifolii were leaf blights, dying from the leaf tip downwards to the crown, which appeared patches in the field because of local pockets of severely infected (blighted) grass. Perithecia were produced on old or weak leaves, including club-shaped asci, each of which contained 8 pale brown muriform ascospores with cross and longitudinal septa. Ascospores of the fungus isolated from the diseased leaf tissue and cultured on potato-dextrose agar (PDA) were muriform multicellular (composed of 3-6 cells) and $23.4-40.5{\times}7.8-15.6{\mu}m$ in size with 3-4 transverse and 0-3 longitudinal septa, which were morphologically identical to L. trifolii reported previously. DNA sequences of ribosomal RNA gene (internal transcribed spacer) of the fungus were homologous with similarity of 99% to those of L. trifolii isolates in GenBank database, confirming the identity of the causal agent of the disease. Pathogenicity of the fungus was also confirmed on the creeping bentgrass by Koch's postulates. This is first report of Leptosphaerulina leaf blight on turfgrass caused by L. trifolii in Korea.

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.317-323
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• 2014

Violacein has received much attention due to its various important biological activities, including broad-spectrum antibacterial and antifungal activity, anti-malarial, anti-tumoral, anti-oxidant, and anti-diarrheal activities. EP15224 strain isolated from forest soils in Korea was found to be a new species belonged to the genus Massilia based on its 16S ribosomal DNA sequences. The 16S ribosomal DNA of strain EP15224 displayed 97% homology with Massilia sp. BS-1, the nearest violacein-producing bacterium. Strain EP15224 produced bluish-purple pigment well in a synthetic MM2 medium containing glucose, $(NH_4)_2SO_4$, $Na_2HPO_4{\cdot}7H_2O$, $KH_2PO_4$, $MgSO_4{\cdot}7H_2O$, and 1 mM $\small{L}$-tryptophan. The chemical analysis of the pigment by LC/MS/MS showed that it is violacein with molecular weight of 343.34. This is the second report on the production of violacein by a Massilia species. In this study, the optimal culture conditions for violacein production were established under which 280 mg/l crude violacein was produced : glucose 2 g/l, $(NH_4)_2SO_4$ 1 g/l, $Na_2HPO_4{\cdot}7H_2O$ 2 g/l, $KH_2PO_4$ 1 g/l, $MgSO_4{\cdot}7H_2O$ 0.1 g/l, L-tryptophan 0.24 g/l, 25 ml medium in a 250 ml flask, with an inoculumn size of 10% (v/v), 72 h of cultivation with 250 rpm at $25^{\circ}C$.

• Journal of Life Science
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• v.29 no.1
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• pp.84-89
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• 2019

Previous screening of novel antibacterial agents revealed that some bacterial isolates exhibited antibiotic activity against both gram-positive and gram-negative bacteria and that they showed antibacterial activity, even against methicillin-resistant Staphylococcus aureus (MRSA). Among these isolates, one bacterial strain, BCNU 1204, was identified as Pseudomonas aeruginosa using phenetic and phylogenetic analysis, based on 16S ribosomal RNA gene sequences. The maximum productivities of antimicrobial substances of BCNU 1204 were obtained after being cultured at $35^{\circ}C$ and pH 7.0 for 4 d in King's medium B (KMB). Dichloromethane (DCM) and ethylacetate (EA) extracts of P. aeruginosa BCNU 1204 exhibited strong antimicrobial activity, particularly against gram-positive bacteria. The EA extracts exhibited broad-spectrum activity against antibiotic resistant strains. Fraction 5-2, was obtained by recycling preparative liquid chromatography (LC) and preparative thin-layer chromatography (TLC) and was identified as phenazine-1-carboxylic acid belonging to phenazines using gas chromatography and mass spectrometry (GC/MS). Its minimum inhibitory concentration (MIC) values were $25{\mu}g/ml$, $50{\mu}g/ml$, ${\geq}25{\mu}g/ml$, and ${\geq}50{\mu}g/ml$ for MRSA CCARM 3089, 3090, 3091, and 3095 strains, respectively. P. aeruginosa BCNU 1204 may be a potential resource for the development of anti-MRSA antibiotics. Additional research is required to identify the active substance from P. aeruginosa BCNU 1204.

• Journal of Veterinary Clinics
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• v.28 no.5
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• pp.516-518
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• 2011

A survey was conducted to examine the normal intestinal bacterial flora of captive Oriental white storks (Ciconia boyciana) maintained at the Korea Institute of Oriental White Stork Rehabilitation Research, Cheongwon, South Korea. From the cloaca of 44 healthy storks, 44 fecal samples were collected and cultured under aerobic and anaerobic conditions. The 16S ribosomal RNA gene sequences and the heat shock protein 60 gene were cloned and sequenced for bacterial identification. Under aerobic conditions, Enterococcus faecalis, Escherichia coli, Bacillus spp., Enterococcus avium, Enterococcus gallinarum, Pseudomonas spp., Alcaligenes spp., Enterobacter spp., Corynebacterium spp., and Proteus mirabilis were identified. Under anaerobic conditions, E. coli, Clostridium tertium, En. faecalis, and P. mirabilis were identified. E. coli, En. faecalis, or both were isolated from all samples. These results will add to the information available on this stork species and help for the interpretation of fecal culture results.

• Research in Plant Disease
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• v.24 no.4
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• pp.342-347
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• 2018

Bacterial canker caused by Clavibacter michiganensis is considered to be one of the most serious diseases, leading to economic damage to tomato worldwide. Diagnosis of the bacterial canker on tomato is known to be difficult because the causal pathogen is slow-growing on artificial media as well as causes latent infection in tomato. In this study, as a less time-consuming method, a specific primer set was newly designed for rapid detection of C. michiganensis. The method presented here is so simple, easy, and fast that it can be useful and practical in direct detection of the bacterial canker pathogen from tomato plants.

• Journal of Nutrition and Health
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• v.56 no.4
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• pp.349-360
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• 2023

Purpose: Ketogenic diets (KDs) have anti-obesity effects that may be related to glucose control and the gut microbiota. This paper hypothesizes that KD reduces body weight and changes the insulin sensitivity and gut microbiota composition in a mouse model of diet-induced obesity. Methods: In this study, C57BL/6 male mice were assigned randomly to 3 groups. The assigned diets were provided to the control and high-fat (HF) diet groups for 14 weeks. The KD group was given a HF diet for 8 weeks to induce obesity, followed by feeding the KD for the next 6 weeks. Results: After the treatment period, the KD group exhibited a 35.82% decrease in body weight gain compared to the HF group. In addition, the KD group demonstrated enhanced glucose control, as shown by the lower levels of serum fasting glucose, serum fasting insulin, and the homeostatic model assessment of insulin resistance, compared to the HF group. An analysis of the gut microbiota using 16S ribosomal RNA sequencing revealed a significant decrease in the proportion of Firmicutes when the KD was administered. In addition, feeding the KD reduced the overall alpha-diversity measures and caused a notable separation of microbial composition compared to the HF diet group. The KD also led to a decrease in the relative abundance of specific species, such as Acetatifactor_muris, Ligilactobacillus_apodemi, and Muribaculum_intestinale, compared with the HF group. These species were positively correlated with the body weight, whereas the abundant species in the KD group (Kineothrix_alysoides and Saccharofermentans_acetigenes) showed a negative correlation with body weight. Conclusion: The current study presents supporting evidence that KD reduced the body weight and altered the insulin sensitivity and gut microbiota composition in a mouse model of diet-induced obesity.

• Journal of Veterinary Clinics
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• v.31 no.3
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• pp.194-198
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• 2014

Methicillin-resistant staphylococci (MRS) are emerging as important pathogens in humans and animals worldwide. The aim of this study was to investigate the prevalence of MRS in the racehorse population and in horse-related personnel in Korea. A total of 195 horses and 18 humans (eight veterinarians, three veterinary hospital staff, and seven horse-handlers) from racehorse farms in Korea were included in the study. The samples were collected from nasal cavities using bacterial transport medium and were cultivated on tryptic soy agar with 5% sheep blood for 3 days at $37^{\circ}C$ to confirm the presence of Staphylococcus spp. Presumptive Staphylococcus spp. isolates were identified by 16S ribosomal RNA gene analysis. The coagulase test and oxacillin susceptibility tests were performed using the tube dilution and disk diffusion methods, respectively. The presence of the mecA gene was determined using a polymerase chain reaction assay. Of the 195 horses, 29 (15.6%) yielded 29 MRS isolates. Twelve (66.7%) of the 18 horse-related personnel yielded 12 MRS isolates. All of the MRS isolates from horses or horse-related personnel were identified as methicillin-resistant coagulase-negative staphylococci (MRCNS). The result of this study suggest that the prevalence of MRS increased with the duration of antibiotic use (p = 0.002). This study also provides evidence for the zoonotic transmission of MRCNS between horses and humans, although further investigations are needed.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.18
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• pp.8281-8285
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• 2016

Background: Helicobacter pylori is a cause of chronic gastritis, peptic ulcer disease, and gastric malignancy, infection being a serious health problem in Thailand. Recently, clarithromycin resistant H. pylori strains represent the main cause of treatment failure. Therefore this study aimed to determine the prevalence and pattern of H. pylori resistance to clarithromycin in Suranaree University of Technology Hospital, Suranree University of Technology, Nakhon Ratchasima, Northeastern Thailand, Nakhon Ratchasima province, northeast of Thailand. Materials and Methods: This hospital-based cross-sectional study was carried out between June 2014 and February 2015 with 300 infected patients interviewed and from whom gastric mucosa specimens were collected and proven positive by histology. The gastric mucosa specimens were tested for H. pylori and clarithromycin resistance by 23S ribosomal RNA point mutations analysis using real-time polymerase chain reactions. Correlation of eradication rates with patterns of mutation were analyzed by chi-square test. Results: Of 300 infected patients, the majority were aged between 47-61 years (31.6%), female (52.3%), with monthly income between 10,000-15,000 Baht (57%), and had a history of alcohol drinking (59.3%). Patient symptoms were abdominal pain (48.6%), followed by iron deficiency anemia (35.3%). Papaya salad consumption (40.3%) was a possible risk factor for H. pylori infection. The prevalence of H. pylori strains resistant to clarithromycin was 76.2%. Among clarithromycin-resistant strains tested, all were due to the A2144G point mutation in the 23S rRNA gene. Among mutations group, wild type genotype, mutant strain mixed wild type and mutant genotype were 23.8%, 35.7% and 40.5% respectively. With the clarithromycin-based triple therapy regimen, the efficacy decreased by 70% for H. pylori eradication (P<0.01). Conclusions: Recent results indicate a high rate of H. pylori resistance to clarithromycin. Mixed of wild type and mutant genotype is the most common mutant genotype in Nakhon Ratchasima province, therefore the use of clarithromycin-based triple therapy an not advisable as an empiric first-line regimen for H. pylori eradication in northeast region of Thailand.