• KSBB Journal
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• v.28 no.5
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• pp.295-302
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• 2013

Indigo is utilized in various industries including textile dyeing, cosmetics, printing and medicinal products and its reduced form, leuco-indigo, is mainly used in these process. Chemical reducing agent (sodium dithionite, sodium sulfide, etc.) is preferred to use for the formation of leucoindigo in industry. In traditional indigo fermentation process, microorganisms can participate in the reduction of indigo and thus it has been known to reduce environmental pollution and noxious byproducts. However, in fermentation method using microorganisms it is difficult to standardize large scale production process due to low yield and reproducibility. In this study, we attempted to develop the indigo reduction process using microbial flora which was isolated from naturally fermented indigo vat or deduced by metagenomic approach. From the results of library analyses of PCR-amplified 16S rRNA genes from the traditional indigo fermentation vat sample (metagenome), it was confirmed that Alkalibacteriums (71%) was distinctly dominant in population. Some strains were identified after confirming that they become pure culture in nutrient media modified slightly. Four strains were separated in this process and each strain showed obvious reducing ability toward indigo in dyeing test. It is expected that the analyzed results will provide important data for standardizing the natural fermentation of indigo and investigating the mechanism of indigo reduction.

• Korean Journal of Veterinary Service
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• v.35 no.2
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• pp.99-104
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• 2012

Otitis externa (OE) is a frequent disease in the ear canals of dogs. To identify the pathogens causing OE in dogs and to determine their antimicrobial resistances, specimens were collected from animal hospitals in Daejeon. The isolates were examined by morphological and biochemical tests, 16S rRNA analysis and antimicrobial susceptibility tests. We analyzed correlation between the isolated pathogens and external factors of dogs such as breed, age, gender, ear mite, hair in ears and experience with antibiotic therapy. Thirty three strains of bacteria were isolated from 26 of the 68 heads of dogs with OE. The most isolated bacteria were Enterococcus faecalis (E. faecalis) followed by Staphylococcus aureus (Sta. aureus), Sta. pseudointermedius, E. faecium, E. avium and Streptococcus canis (Strep. canis) in order of frequency of occurrence. Isolation frequency of Enterococcus spp. and Staphylococcus spp. were 51.5% and 45.5%, respectively. E. faecalis and E. faecium isolates showed VanB phenotype, which is resistant to vancomycin but sensitive to teicoplanin were 58% and 25%, respectively. Nine isolates among total twelve isolates of E. faecalis were isolated from the dogs treated with antibiotics. There was no methicillin-resistant Sta. aureus (MRSA), but were MR-Sta. pseudointermedius (MRSP) (57.1%) and vancomycin-resistant (VR)-Sta. pseudointermedius (14.3%) (VRSP) showing VanB phenotype. However, vanA, vanB and vanC genes were not detected in VR isolates from the dogs. Taken together, VR-Enterococcus spp. (VRE) is one of the major pathogens in domestic animals, as well as community-and hospital-acquired infection.

• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.784-791
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• 2007

The present work aims to use a biofilter technology(aerated submerged filters) for the aerobic transformation at laboratory-scale of olive washing water(OWW) generated in the first steps of olive oil processing, as well as the genetic profiling and identification to the species level of the bacteria involved in the formation of the biofilm, by means of TGGE. Chemical parameters, such as biological oxygen demand at five days($BOD_5$) and chemical oxygen demand(COD), decreased markedly(up to 90 and 85%, respectively) by the biological treatment, and the efficiency of the process was significantly affected by aeration and inlet flow rates. The total polyphenol content of inlet OWW was only moderately reduced(around 50% decrease of the inlet content) after the biofilter treatment, under the conditions tested. Partial 16S rRNA genes were amplified using total DNA extracted from the biofilm and separated by TGGE. Sequences of isolated bands were mostly affiliated to the $\alpha-subclass$ of Proteobacteria, and often branched in the periphery of bacteria] genera commonly present in soil(Rhizobium, Reichenowia, Agrobacterium, and Sphingomonas). The data obtained by the experimentation at laboratory scale provided results that support the suitability of the submerged filter technology for the treatment of olive washing waters with the purpose of its reutilization.

• Korean Journal of Veterinary Research
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• v.51 no.2
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• pp.107-115
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• 2011

Theileria (T.) buffeli (formerly T. sergenti/T. orientalis) is the major hemo-protozoan distributed in the Far East Asian countries such as Korea, China and Japan. It is responsible for the clinical symptoms of anorexia, ateliosis, anemia, fever and icterus. It also causes abortion and sudden death under severe cases, resulting in economic losses for many livestock farms. The objective of this study was to analyze the genetic diversity of the major surface protein (Msp) gene in T. buffeli in Holstein in Korea, and we characterized the association of the diversification of the Msp gene and its relationship with the pathogenicity of Theileria. For this, complete blood counts and Theileria PCR sequence analysis were performed from 57 Holstein in Jeju Island. A total of 26 PCR positive Holstein (16 anemic and 10 non-anemic) were then randomly selected based on 18s rRNA sequence typing of the Theileria Msp gene. The DNA sequence of the T. buffeli Msp gene in Holstein showed 99.0%, 99.2%, 99.9%, 99.5%, 98.7%, 98.4% and 98.4% homology with T. sergenti, Theileria spp., T. sergenti, Theileria spp., Theileria spp., Theileria spp. and Theileria spp., respectively. The result showed a genetic variation of 57.7% (type I), 3.8% (type II), 15.4% (type III), 7.7% (type IV), 13.5% (type V) and 1.9% (type VI). Type I is the most frequent type in both anemic and non-anemic Holstein while type II was found in only non-anemic Holstein. This results of our study help confirm the diversity of Msp gene types and demonstrate that the gene type distribution of Msp genes varies among Theileria-infected Holstein in Jeju Island.

• Journal of Animal Science and Technology
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• v.45 no.5
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• pp.695-702
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• 2003

Nucleolus Organizer Regions (NORs) are the specific chromosome sites where ribosomal genes are located and highly expressed. We have applied the AgNOR staining to identify the distribution of NORs in the chromosomes of Korean Cattle. We have also studied the NORs pattern on the cells originated from different breeds, tissues and sex. Peripheral blood from forty-four Korean Cattle and Holstein was cultured for chromosme preparation. The fibroblast culture from biopsied ear skins was also conducted for chromosome analysis. The distribution of NORs was analyzed by sequential Ag staining and G-banding on metaphases of the cells. In Korean Cattle, the NORs are localized on the telomeres of the five chromosome pairs number 2, 3, 4, 11 and 28. The number of NORs per metaphase ranged from 2 to 10 giving a mean value of 5.6. The number of NORs per cell varied among individuals and cells within same individual. The size of NORs also differed in NO-chromosomes. The number of NORs was significantly different between Korean Cattle and Holstein, fibroblasts and lymphocytes, and male and female. However, the distribution and frequency of NORs were similar among the cells regardless of breeds, tissues, and sex.

• Development and Reproduction
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• v.8 no.1
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• pp.49-55
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• 2004

Aquaporins(AQPs) are a family of transmembrane water channel proteins that are widely distributed in various tissues throughout the body and play a major role in Oanscellular and Oansepithelial water movement. Uterine endometrium undergoes recurrent uterine stromal edema in response to hormonal stimuli, however, the mechanism regulating the fluid transport during the estrous cycle has not been fully understood. To investigate the possible role of AQPs in water movement in uterus during the estrous cycle, expression patterns of AQP -1, -3, -4, -5, -8, and -9 UMh in mouse uterus were analyzed by using semiquantitative reverse transcription- polymerase chain reaction(RT-nR). We employed a combination of laser capture microdissection(LCM) and RT-PCR to examine the expression patterns in specific uterine cell types luminal epithelial cells(LE) and stromal cells(S). Our results showed that the level of AQP-4 mRNA was significantly increased while the level of AQP-3 mRNA was significantly decreased during the proestous through the estrus stage. In addition LCM revealed that AQP-4 and -8 mRNAs were highly expressed in LE compared with S. Taken together, these results suggest that AQPs may have an important function in physiological changes of mouse uterus during the estrous cycle.

• Research in Plant Disease
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• v.11 no.2
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• pp.140-145
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• 2005

A collection of 12 Erwinia carotovora strains from blackleg diseased potato in Jeju island was characterized genetic diversity by 5. cayotovora subsp. atposeptica (Eca)-specific PCR, PCR-RFLP of the two genes (16S rRNA and pel) and repetitive sequence PCR (ERIC-PCR). The results were compared with those of the other E. carotovora representative strains. None of the blackleg strains produced PCR amplicons with Eca-specific primers in contrast to the single 690 bp amplicon obtained with Eca strains. In addition, on the basis of pel gene RFLP with Sau3AI, the blackleg strains belonged to the pattern 2 whereas Eca strains belonged to the other one (pattern 3). By analysis of 16S rDNA RELP generated with HinfI, the most strains including the E. carotovera subsp. carotovora (Ecc) representative strains used in this study belonged to the pattern 1 whereas the blackleg strains belonged to the pattern 2 except for one strain. Moreover, ERIC-PCR analysis showed that the blackleg strains were closely related to each other and had an unique DNA band. Based on these molecular approaches, we have confirmed that the blackleg disease of potato is caused by a different E. carotovora from Eca and Ecc in Jeju island.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.6
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• pp.671-680
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• 2017

This study was conducted to examine the effects and potential mechanisms of action of black soybean extracts and fermented black soybean extracts by Lactobacillus rhamnosus GG (LGG) and Bifidobacterium animals subsp. lactis BB-12 (BB-12) on proliferation of human follicle dermal papilla cells (HFDPC). We examined changes in pH, total polyphenol, sugar, and reducing sugar contents according to fermentation period of black soybean extracts. Assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was performed to determine cell toxicity levels of the four black soybean extracts [black soybean water extract (BWE), black soybean ethanol extract (BEE), fermented BWE (F-BEW), and fermented BEE (F-BEE)]. Changes in mRNA expression levels of hair growth promoting factors and hair growth inhibiting factors by the four black soybean extracts were measured by real-time PCR. In addition, phosphorylation levels of mitogen-activated protein kinase family proteins were measured by western blot analysis. As a result, fermentation of black soybeans significantly reduced pH, total polyphenols, and sugar/reducing sugar contents. All four black soybean extracts showed no cellular toxicity in HFDPC. In fact, BEE significantly enhanced cell viability of HFDPC at $100{\mu}g/mL$ compared to control. BWE, BEE, and BWE-F significantly increased mRNA expression of vascular endothelial growth factor, and all four extracts increased mRNA expression of fibroblast growth factor. However, mRNA expression levels of apoptosis-related genes were not affected by black soybean extracts in HFDPC. Furthermore, BWE, BEE, and BWE-F significantly increased phosphorylation levels of extracellular signal-regulated kinase compared to control. Taken together, we demonstrated that black soybean extracts enhanced proliferation of human follicle dermal papilla cells partially via activation of hair growth promoting factors, although no particular significant effects on proliferation were observed by fermentation of black soybeans.

• Journal of Microbiology and Biotechnology
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• v.24 no.2
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• pp.270-279
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• 2014

Microbial community shifts, associated with performance data, were investigated in an anaerobic batch digester treating high-solid food waste under mesophilic conditions using, a combination of molecular techniques and chemical analysis methods. The batch process was successfully operated with an organic removal efficiency of 44.5% associated with a biogas yield of 0.82 L/g $VS_{removal}$. Microbial community structures were examined by denaturing gel gradient electrophoresis. Clostridium and Symbiobacterium organisms were suggested to be mainly responsible for the organic matter catabolism in hydrolysis and acidogenesis reactions. The dynamics of archaeal and methanogenic populations were monitored using real-time PCR targeting 16S rRNA genes. Methanosarcina was the predominant methanogen, suggesting that the methanogenesis took place mainly via an aceticlastic pathway. Hydrogenotrophic methanogens were also supported in high-solid anaerobic digestion of food waste through syntrophism with syntrophic bacterium. Microbial community shifts showed good agreement with the performance parameters in anaerobic digestion, implying the possibility of diagnosing a high-solid anaerobic digestion process by monitoring microbial community shifts. On the other hand, the batch results could be relevant to the start-up period of a continuous system and could also provide useful information to set up a continuous operation.

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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• pp.354-359
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• 2003

Multiplex-PCR protocols were designed in order to make a rapid identification of MRSA. MecA, femB, and 165 rRNA genes were amplified for making a detection of MRSA. The incidence of MRSA in the clinical isolates of Staphylococcus aureus was examined by using a multiplex-PCR assay. The mecA gene was detected in 266 strains out of 336 clinical isolates of S. aureus, thus the incidence of MRSA was approximately 76.5%. The MRSAs of 247 strains (96.1%) showed resistance to more than eight species of the antimicrobial agents tested. The isolates of MRSA showed 27 different antimicrobial-resistant patterns. The results indicate that many different MRSA strains having high multidrug resistance are actually prevalent in Korea. Also, VISA was screened from the MRSA. Two strains were grown on the BHI agar plate supplemented with $8\;\mu\textrm{g}/ml$ of vancomycin at a frequency of $1/10^8$ colony forming units or higher.