• Title/Summary/Keyword: 28S rRNA gene

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Isolation and Characterization of Marine Bacterial Strain SH-1 Producing Agar-Degrading Enzymes (한천 분해효소를 생산하는 해양 미생물 SH-1의 분리 및 특성 분석)

  • Lee, Jae-Hag;Lee, Soon-Youl
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.324-330
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    • 2014
  • A marine bacterial strain producing agar-degrading enzymes was isolated from a mud flat in Jeboo-do (Korea) using a selective artificial sea water (ASW) agar plate containing agar as the sole carbon source. The isolate, designated as SH-1, was gram-negative, aerobic, and motile with single polar flagellum. 16S rRNA gene sequence similarity analysis showed the isolate SH-1 had the highest homology (96.5%) to marine bacterium Neiella marina J221. Cells could grow at $28-37^{\circ}C$ but not at $42^{\circ}C$, and the agarase activity of the cell culture supernatant was higher when grown at $28^{\circ}C$ than when grown at $37^{\circ}C$. Cells could grow when concentrations of 1-5% (w/v) NaCl were added to the growth media with the best growth observed at 3% NaCl, and the agardegrading enzyme activity of the cell culture supernatant was best when grown at 3% NaCl-containing growth media under the conditions we examined. The crude enzyme prepared from 48-h culture broth of strain SH-1 exhibited an optimum pH and temperature for agar-degrading activity at 7.0 and $40^{\circ}C$, respectively. Zymogram analysis of the crude supernatant and cell extract showed that strain SH-1 produced at least 3 agar-degrading enzymes with molecular weights of 15, 35, and 52 KD. Thinlayer chromatography (TLC) analysis also suggested that HS-1 produces ${\beta}$-agarase to degrade agarose to neoagarooligosaccharides.

Effects of Immune Stress on Performance Parameters, Intestinal Enzyme Activity and mRNA Expression of Intestinal Transporters in Broiler Chickens

  • Feng, Y.;Yang, X.J.;Wang, Y.B.;Li, W.L.;Liu, Y.;Yin, R.Q.;Yao, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.701-707
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    • 2012
  • Immune stress is the loss of immune homeostasis caused by external forces. The purpose of this experiment was to investigate the effects of immune stress on the growth performance, small intestinal enzymes and peristalsis rate, and mRNA expression of nutrient transporters in broiler chickens. Four hundred and thirty-two 1-d-old broilers (Cobb500) were randomly assigned to four groups for treatment; each group included nine cages with 12 birds per cage. Group 1 = no vaccine (NV); Group 2 = conventional vaccine (CV); group 3 = lipopolysaccharide (LPS)+conventional vaccine (LPS); group 4 = cyclophosphamide (CYP)+conventional vaccine (CYP). The results demonstrated that immune stress by LPS and CYP reduced body weight gain (BWG), feed intake (FI), small intestine peristalsis rate and sIgA content in small intestinal digesta (p<0.05). However, the feed conversion ratio (FCR) remained unchanged during the feeding period. LPS and CYP increased intestinal enzyme activity, relative expression of SGLT-1, CaBP-D28k and L-FABP mRNAs (p<0.05). LPS and CYP injection had a negative effect on the growth performance of healthy broiler chickens. The present study demonstrated that NV and CV could improve growth performance while enzyme activity in small intestine and relative expression of nutrient transporter mRNA of NV and CV were decreased in the conditions of a controlled rational feeding environment. It is generally recommended that broilers only need to be vaccinated for the diseases to which they might be exposed.

Identification of Red Tide-causing Organism and Characteristics of Red Tide Occurrence in the Oncheon Down Stream, Busan (온천천 하류 적조 원인생물의 동정 및 발생 특성)

  • Kim, Mi-hee;Ji, Hwa-seong;Cho, Jeong-goo;Cho, Sunja
    • Journal of Korean Society on Water Environment
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    • v.34 no.3
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    • pp.285-292
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    • 2018
  • This study was performed in order to identify the red tide-causing organism and to understand the characteristics of the water quality during the winter of 2015 and 2016 in the Oncheon stream, a tidal river in Busan, where red tide often occurs in the wintertime. Two sites were selected on the stream and the surface water was sampled a total of 28 times during the experimental period. Twelve water quality characteristics, including water temperature, pH, DO, COD, total-N (T-N), total-P (T-P), and salinity were analyzed in order to test water quality. The cell numbers of cryptomonads were counted directly by microscopic observation. The nucleotide sequences of the partial 28S rRNA gene and psbA gene from metagenomic DNA, derived from each sampling site, were analyzed. According to the results, the alga most responsible for the bloom was identified as Teleaulax OC1 sp., which belongs to the cryptomonads. Three items of chl-a, pH, and DO were positively correlated with the cell numbers of the cryptomonads counted at the upper stream of the tidal area (St 1) while eight items of chl-a, TOC, BOD, total-N, COD, SS, pH, and DO were positively correlated with the cells located at the junction between the stream and Su-young river (St 2) in the order.

Diversity and physiological properties of soil actinobacteria in Ulleung Island (울릉도 유래 토양 방선균의 다양성과 생리활성)

  • Yun, Bo-Ram;Roh, Su Gwon;Kim, Seung Bum
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.242-250
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    • 2017
  • Actinobacteria tolerating extreme conditions can be a rich source of bioactive compounds and enzymes. In this study filamentous actinobacteria were isolated from soils of Ulleung Island, and their physiological properties were examined. Soil samples were collected, serially diluted and spread on various agar media. The average viable counts of total bacteria were $1.28{\times}10^7CFU/g$ for soil sample 1 (ULS1) and $2.05{\times}10^7CFU/g$ for soil sample 2 (ULS2). As a result, 34 strains of actinobacteria were isolated and assigned to the genera Streptomyces (16 strains), Isoptericola (5 strains), Rhodococcus (4 strains), Agromyces (3 strains), Micrococcus (2 strains), Arthrobacter (1 strain), Williamsia (1 strain), Microbacterium (1 strain), and Oerskovia (1 strain) based on 16S rRNA gene sequence analysis. Enzyme activity and plant growth promoting potential were tested for representative isolates. Multiple strains of Streptomyces degraded starch, casein and Tween 80. As for plant growth promoting potential, strains of Oerskovia, Williamsia, Isoptericola, and Streptomyces solubilized phosphate, and those of Agromyces, Oerskovia, Micrococcus, Rhodococcus, Streptomyces, and Isoptericola produced 3-indole-acetic acid (IAA), respectively. Selected strains of Streptomyces exhibited strong antagonistic activity against Staphylococcus aureus and Bacillus subtilis as well as Candida albicans. This study confirms that actinobacteria from Ulleung Island can be a good source of novel bioactive compounds.

Superoxide Dismutase Gene Expression in the Endotoxin-Treated Rat Lung (내독소에 의한 백서 폐장의 Superoxide Dismutase 유전자 발현에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.3
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    • pp.215-221
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    • 1994
  • Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

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Association of Salivary Microbiota with Dental Caries Incidence with Dentine Involvement after 4 Years

  • Kim, Bong-Soo;Han, Dong-Hun;Lee, Ho;Oh, Bumjo
    • Journal of Microbiology and Biotechnology
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    • v.28 no.3
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    • pp.454-464
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    • 2018
  • Salivary microbiota alterations can correlate with dental caries development in children, and mechanisms mediating this association need to be studied in further detail. Our study explored salivary microbiota shifts in children and their association with the incidence of dental caries with dentine involvement. Salivary samples were collected from children with caries and their subsequently matched caries-free controls before and after caries development. The microbiota was analyzed by 16S rRNA gene-based high-throughput sequencing. The salivary microbiota was more diverse in caries-free subjects than in those with dental caries with dentine involvement (DC). Although both groups exhibited similar shifts in microbiota composition, an association with caries was found by function prediction. Analysis of potential microbiome functions revealed that Granulicatella, Streptococcus, Bulleidia, and Staphylococcus in the DC group could be associated with the bacterial invasion of epithelial cells, phosphotransferase system, and ${\text\tiny{D}}-alanine$ metabolism, whereas Neisseria, Lautropia, and Leptotrichia in caries-free subjects could be associated with bacterial motility protein genes, linoleic acid metabolism, and flavonoid biosynthesis, suggesting that functional differences in the salivary microbiota may be associated with caries formation. These results expand the current understanding of the functional significance of the salivary microbiome in caries development, and may facilitate the identification of novel biomarkers and treatment targets.

Flavobacterium amnigenum sp. nov. Isolated from a River

  • Patil, Kishor Sureshbhai;Padakandla, Shalem Raj;Chae, Jong-Chan
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1536-1541
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    • 2018
  • A yellowish, flexirubin-pigment-producing strain $I3-3^T$ was isolated from river water in Iksan, the Republic of Korea. The strain was gram-negative, aerobic, non-motile, showed catalase and oxidase activities, and could grow at a temperature range of $10-35^{\circ}C$, pH 5.0-10 and 0-2.0% (w/v) of NaCl. The major fatty acids were iso-$C_{15:0}$, iso-$C_{17:0}$ 3-OH and summed feature 3 (comprising $C_{16:1}{\omega}7c$ and/or $C_{16:1}{\omega}6c$). The isolate contained phosphatidylethanolamine, one aminolipid, and two unidentified lipids as the major polar lipids. Menaquinone-6 (MK6) was the major respiratory quinone. The G+C content of the genomic DNA of strain $I3-3^T$ was 35.6%. Comparison of the 16S rRNA gene sequence with the sequences of the closely related type strains showed highest sequence similarity of 96.95% and 96.93% to Flavobacterium nitrogenifigens $NXU-44^T$ and Flavobacterium compostarboris $15C3^T$, respectively. Based on phenotypic and phylogenetic distinctiveness, strain $I3-3^T$ is considered as a member of novel species within the genus Flavobacterium, for which Flavobacterium amnigenum sp. nov. is proposed. The type strain is $I3-3^T$ (=KCTC $52884^T$ =NBRC $112871^T$).

First Report of Plectosphaerella sinensis Isolated from Soil in Korea

  • Wajihi, Ally Hassan;Choi, Hyo-Won;Lee, Seung-Yeol;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.46 no.3
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    • pp.345-351
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    • 2018
  • A fungal isolate denoted NC14-264 was isolated from the soil in Jeongeup, Jeollabuk-do, Korea. Species in genus Plectosphaerella are pathogenic to several plant species, leading to fruit, root, and collar rot and collapse. In this study, a strain NC14-264 belonging to the Plectosphaerella was isolated from the soil and identified. Colonies were moderately grown, reaching 54 mm in diameter on potato dextrose agar, 49 mm on malt extract agar, and 55 mm on oatmeal agar at $25^{\circ}C$ after 10 days of incubation. Most Plectosphaerella species are distinguishable morphologically by irregular chlamydospores and different proportions of phialides and conidia. Based on morphological features and phylogenetic analysis using the internal transcribed spacer region and partial 28S rRNA gene sequences, the isolated fungus was identified as Plectosphaerella sinensis belonging to the Plectosphaerellaceae. This is the first report of P. sinensis in Korea.

The First Korean Case of Nontuberculous Mycobacterial Lung Disease Caused by Mycobacterium abscessus Subspecies bolletii in a Patient with Bronchiectasis

  • Jeong, Byeong-Ho;Kim, Su-Young;Jeon, Kyeongman;Huh, Hee Jae;Ki, Chang-Seok;Lee, Nam Yong;Shin, Sung Jae;Koh, Won-Jung
    • Tuberculosis and Respiratory Diseases
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    • v.76 no.1
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    • pp.30-33
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    • 2014
  • We report the first Korean case of lung diseases caused by Mycobacterium abscessus subsp. bolletii in a previously healthy male, except for a previous history of pulmonary tuberculosis and bronchiectasis. All serial isolates are identified as M. abscessus subsp. bolletii by multi-locus sequence analysis based on the hsp65, rpoB, and 16S rRNA fragments. At the genetic level, the isolate has the erm(41) gene with a T28 sequevar, associated with clarithromycin resistance, and no rrl mutation. The isolate is resistant to clarithromycin. Although the symptoms and radiographic findings have improved after combination of antibiotics, the follow-up sputum cultures are persistently positive.

Characterization of the Thermophilic Bacterium Geobacillus sp. Strain GWE1 Isolated from a Sterilization Oven

  • Correa-Llanten, Daniela;Larrain-Linton, Juanita;Munoz, Patricio A.;Castro, Miguel;Boehmwald, Freddy;Blamey, Jenny M.
    • Microbiology and Biotechnology Letters
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    • v.41 no.3
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    • pp.278-283
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    • 2013
  • A gram-positive, rod-shaped, spore-forming, motile thermophilic bacterium was isolated from a sterilization oven. The microorganism GWE1, formally named Geobacillus wiegelii identified as a member of the genus Geobacillus. GWE1 grew under aerobic conditions of between $60-80^{\circ}C$ (optimum $670^{\circ}C$), in a pH range of 3.0-8.0 (optimum $pH^{70^{\circ}C}$ 5.8), and between 0 and 2 M NaCl (optimum 0.3 M). The membrane polar lipids were dominated by branched saturated fatty acids, which included as the major constituents; iso-15:0 (13.3%), 16:1(${\omega}7$) (12.8%), 16:0 (28.5%), iso-17:0 (13.5%) and anteiso-17:0 (12.3%). The DNA G+C content was 47.2 mol% (determined by HPLC). The 16S rRNA gene sequence of GWE1 showed a high similarity with Geobacillus caldoxylosilyticus (97%). However, the level of DNA-DNA relatedness was only 58%. These data suggest that GWE1 is probably a novel specie of the genus Geobacillus.