• Title/Summary/Keyword: 16S ribosomal RNA

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Occurrence of Leptosphaerulina Leaf Blight on Kentucky Bluegrass Caused by Leptosphaerulina trifolii (Leptosphaerulina trifolii에 의한 Kentucky Bluegrass의 Leptosphaerulina 잎마름병 발생)

  • Kim, Jeong-Ho;Shim, Gyu-Yul;Kim, Young-Ho
    • 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.

Production of Violacein by a Novel Bacterium, Massilia sp. EP15224 Strain (Violacein을 생산하는 Massilia sp. EP15224 균주)

  • Yoon, Sang-Hong;Baek, Hee-Jin;Kwon, Soon-Wu;Lee, Chang-Muk;Sim, Joon-Soo;Hahn, Bum-Soo;Koo, Bon-Sung
    • 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 $25^{\circ}C$.

Antimicrobial Activity of Pseudomonas aeruginosa BCNU 1204 and Its Active Compound (Pseudomonas aeruginosa BCNU 1204의 항균활성과 활성 물질)

  • Shin, Hwa Jin;Joo, Woo Hong
    • 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.

Bacterial Flora of the Intestine in Normal Captive Oriental White Storks (정상적인 사육 황새의 장내 세균총)

  • Han, Jae-Ik;Jang, Hye-Jin;Lee, Sook-Jin;Kang, Hyo-Min;Kim, Su-Kyung;Park, Shi-Ryoung;Na, Ki-Jeong
    • 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.

PCR Detection Method for Rapid Diagnosis of Bacterial Canker Caused by Clavibacter michiganensis on Tomato (토마토 궤양병 신속 진단을 위한 Clavibacter michiganensis의 PCR 검출법)

  • Park, Mi-Jeong;Back, Chang-Gi;Park, Jong-Han
    • 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.

A ketogenic diet reduces body weight gain and alters insulin sensitivity and gut microbiota in a mouse model of diet-induced obesity

  • Sumin Heo;Soo Jin Yang
    • 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.

Prevalence of Methicillin-resistant Staphylococci Isolates from Horses and Horse-related Personnel in Korea (말 및 말관련 종사자의 methicillin 내성 포도상구균의 유병율 조사)

  • Lee, Sang-Kyu;Han, Jae-Ik;Kim, Ill-Hwa;Na, Ki-Jeong;Kang, Hyun-Gu
    • 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.

High Prevalence of Helicobacter pylori Resistance to Clarithromycin: a Hospital-Based Cross-Sectional Study in Nakhon Ratchasima Province, Northeast of Thailand

  • Tongtawee, Taweesak;Dechsukhum, Chavaboon;Matrakool, Likit;Panpimanmas, Sukij;Loyd, Ryan A;Kaewpitoon, Soraya J;Kaewpitoon, Natthawut
    • 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.

Characterization of an Antimicrobial Substance-producing Pseudomonas sp. BCNU 2001 (항생물질을 생산하는 Pseudomonas sp. BCNU 2001 균주의 특성)

  • Yang, Uk-Hee;Choi, Hye-Jung;Ahn, Cheol-Soo;Jeong, Yong-Kee;Kim, Dong-Wan;Joo, Woo-Hong
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.255-262
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    • 2010
  • Strain BCNU 2001 was isolated from soil samples collected from Tea-baek Mountain area. The biochemical characteristics and 16S ribosomal RNA gene sequences of the isolate revealed that the strain belonged to the Pseudomonas aeruginosa. The supernatants had an antimicrobial effect on various kind of bacteria and fungi. Especially BCNU 2001 was able to greatly inhibit the growth of Micrococcus luteus, Proteus mirabilis, Proteus vulgaris, and Aspergillus niger, and its inhibition zone was measured as 18.5 mm against Micrococcus luteus, 19.0mm against Proteus mirabilis, 17.0mm against Proteus vulgaris, and 13.5 mm against Aspergillus niger, respectively. Hexane and dichloromethane extracts of BCNU 2001 exhibited significant activity against bacteria, and dichloromethane and ethylacetate extracts showed significant activity against fungi. Pseudomonas strain BCNU 2001 was also determined to have antimicrobial peptide against various microorganisms including Gram positive bacteria, Gram negative bacteria and fungi. The obtained results may provide preliminary support for the usefulness of Pseudomonas strain BCNU 2001.

Gut-residing Microbes Alter the Host Susceptibility to Autoantibody-mediated Arthritis

  • Lee, Hyerim;Jin, Bo-Eun;Jang, Eunkyeong;Lee, A Reum;Han, Dong Soo;Kim, Ho-Youn;Youn, Jeehee
    • IMMUNE NETWORK
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    • v.14 no.1
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    • pp.38-44
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    • 2014
  • K/BxN serum can transfer arthritis to normal mice owing to the abundant autoantibodies it contains, which trigger innate inflammatory cascades in joints. Little is known about whether gut-residing microbes affect host susceptibility to autoantibody-mediated arthritis. To address this, we fed C57BL/6 mice with water containing a mixture of antibiotics (ampicillin, vancomycin, neomycin, and metronidazol) for 2 weeks and then injected them with K/BxN serum. Antibiotic treatment significantly reduced the amount of bacterial genomic DNA isolated from fecal samples, in particular a gene encoding 16S ribosomal RNA derived from segmented filamentous bacteria. Arthritic signs, as indicated by the arthritic index and ankle thickness, were significantly attenuated in antibiotic-treated mice compared with untreated controls. Peyer's patches and mesenteric lymph nodes from antibiotic-treated mice contained fewer IL-17-expressing cells than those from untreated mice. Antibiotic treatment reduced serum C3 deposition in vitro via the alternative complement pathway. IL-$17^{-/-}$ congenic C57BL/6 mice were less susceptible to K/BxN serum-transferred arthritis than their wild-type littermates, but were still responsive to treatment with antibiotics. These results suggest that gut-residing microbes, including segmented filamentous bacteria, induce IL-17 production in GALT and complement activation via the alternative complement pathway, which cause the host to be more susceptible to autoantibody-mediated arthritis.