• Title/Summary/Keyword: Sequence Analyses

Search Result 936, Processing Time 0.03 seconds

Genetic Identification and Biochemical Characteristics of Edwardsiella Strains Isolated from Freshwater Fishes Cultured in Korea (내수면 양식 어류에서 분리된 Edwardsiella 속 균주들의 유전학적 동정 및 생화학적 특성)

  • Jang, Mun Hee;Kim, Keun-Yong;Lee, Yu Hee;Oh, Yun Kyung;Lee, Jeong-Ho;Song, Jun-Young
    • Journal of fish pathology
    • /
    • v.33 no.2
    • /
    • pp.111-118
    • /
    • 2020
  • The genus Edwardsiella belonging to the family Enterobacteriaceae is a member of Gram-negative rod-shaped bacteria that cause disease in diverse aquatic organisms such as fish, amphibians and reptiles as well as avians and mammals including human throughout the world. This genus had been composed of three species, E. hoshinae, E. ictaluri and E. tarda, but recent researches erected two novel species, E. anguillarum and E. piscicida that were conventionally identified as E. tarda. In this study, we isolated seven strains belonging to the genus Edwardsiella from freshwater fishes that had been reared at inland fish farms in South Korea and investigated their biochemical characteristics and molecular phylogenetic relationships. The seven strains showed typical characteristics of four Edwardsiella species, E. anguillarum, E. ictaluri, E. piscicida and E. tarda, by biochemical analyses of Gram staining, indole and hydrogen sulfide (H2S) production, and API (Analytic Profile Index) 20E test. Molecular phylogenetic analyses inferred from DNA sequence data of both 16S ribosomal RNA (rRNA) and DNA gyrase subunit B (gyrB) genes were congruent with the biochemical characteristics. As a result, both biochemical and molecular phylogenetic analyses identified four strains isolated from three Anguilla species as E. anguillarum, E. piscicida and E. tarda, two strains from Pelteobagrus fulvidraco and Silurus asotus as E. ictaluri, and one strain from Moroco oxycephalus as E. piscicida. In this study, we isolated and successfully identified recently newly erected species, E. anguillarum and E. piscicida in addition to historically notorious pathogenic species, E. ictaluri and E. tarda. In the future study, systematic and comprehensive monitoring of the four Edwardsiella species are required for studying differences in pathogenicity among freshwater fishes.

Genetic Diversity of Xanthomonas arboricola pv. pruni Isolated in Korea (우리나라에서 분리한 Xanthomonas arboricola pv. pruni의 유전적 다양성)

  • Park, So-Yeon;Lee, Young-Sun;Shin, Jong-Sub;Koh, Young-Jin;Jung, Jae-Sung
    • Journal of Life Science
    • /
    • v.19 no.5
    • /
    • pp.684-687
    • /
    • 2009
  • Xanthomonas arboricola pv. pruni, the causal agent of bacterial shot holes in stone fruits, was known to have a low population diversity. To investigate the genetic characteristics of X. arboricola pv. pruni isolated in Korea, three strains which have identical 16S rDNA sequences - including type strain (LMG852), Japanese isolate (MAFF301420) and Korean isolate (XWD1) - were analysed based on the nucleotide sequences of three DNA regions and RAPD pattern. No sequence diversity among the three strains was found within the ITS, glnA and atpD gene sequences. However, five of 756 nucleotides of the atpD gene determined (accession number FJ429319) were different from those of the French strain available from the Genbank database. RAPD analyses performed with 40 different arbitrary primers revealed that two strains isolated from Korea and Japan showed similarity in their band patterns distinguished by type strain. These results suggest that Korean and Japanese strains are very close and belong to a population with a low genetic diversity, and might have a different origin from strains found in West Europe.

Detection of a Large White-Specific Duplication in D-loop Region of the Porcine MtDNA (돼지 mtDNA D-loop 지역의 Large White 특이 중복현상 탐지)

  • Kim, Jae-Hwan;Han, Sang-Hyun;Lee, Sung-Soo;Ko, Moon-Suk;Lee, Jung-Gyu;Jeon, Jin-Tae;Cho, In-Cheol
    • Journal of Life Science
    • /
    • v.19 no.4
    • /
    • pp.467-471
    • /
    • 2009
  • The entire D-loop region of the porcine mitochondrial DNA (mtDNA) was amplified from six pig breeds (Landrace, Duroc, Large White, Korean native pig, Berkshire, and Hampshire) using a primer set designed on the basis of reported porcine mtDNA sequences. From analyses through cloning, DNA sequencing and multiple sequence alignment, an 11-bp (TAAAACACTTA) duplication was observed after known tandem repetition in the D-loop region, which promoted hetroplasmy in mtDNA. Although the existence of the 11-bp duplication has been previously reported in Duroc and Japanese native pigs, there have not been any attempts to know the characteristics of this duplication in other breeds so far. A 150 bp fragment containing the 11-duplication was amplified and typed by polyacrylamide gel electrophoresis (PAGE). All Large Whites had two duplication units and Duroc showed heteromorphic patterns, 11.2% (9/80) of the animals had the 11-bp duplication in total. On the other hand, Landrace, Berkshire, Hampshire and Korean native pigs were non-duplicated. This result showed that the 11-bp duplication could be used as a breed-specific DNA marker for distinguishing pure Landrace and Large White breeds.

Morphology, Phylogeny and Ecology of Hyphomycetes Hyperparasitic to Rusts

  • Park, Mi-Jeong;Park, Jong-Han;Hong, Seung-Beom;Shin, Hyeon-Dong
    • 한국균학회소식:학술대회논문집
    • /
    • 2015.05a
    • /
    • pp.55-55
    • /
    • 2015
  • Rust is one of the most destructive diseases on economically important plants such as agricultural and horticultural crops, as well as forest trees [1]. Chemical treatment is the most effective means to control rust, but use of the chemical fungicides involves inevitable risks to human health and environment [2]. Unfortunately, biocontrol is currently impracticable for rust disease management [3]. It is necessary to exploit biocontrol agents to help prevent rust diseases. As a fundamental research for future development of biocontrol agents for rusts, biodiversity of hyperparasites occurring on rust fungi was investigated. During 2006-2010, 197 fungal isolates of the rust hyperparasites were collected and isolated from various combinations of mycohosts and plant hosts in many regions of Korea. Based on morphological and molecular data, they were identified as 8 genera and 12 species. Besides, phylogenetic relationships between the hyperparasites and related taxa were inferred. A total of 114 isolates of Pseudovirgaria were obtained from rust pustules of Phragmidium spp. and Pucciniastrum agrimoniae infecting rosaceous plants. Phylogenetic analysis using multigene sequences revealed a high level of genetic variability among many isolates of Pseudovirgaria and close correlation between the isolates and mycohosts. Only two species of Pseudovirgaria, P. hyperparasitica and P. grisea are often difficult to distinguish by their morphological similarity, but on the molecular basis they were clearly differentiated from each other. There had been no previous record of P. grisea outside Europe, but the present study has proved its presence in Korea. Among six distinct groups (five of P. hyperparasitica and one of P. grisea) within the Pseudovirgaria isolates, each lineage of P. hyperparasitica was closely associated with specific mycohosts and thus might have cospeciated with their mycohosts, which probably led to coevolution. Although P. grisea possesses a host preference for Phragmidium species occurring on Rubus, it was not specific for a mycohost. P. grisea seems to evolve in the direction of having a broad mycohost range. Seventeen isolates of Verticillium-like fungi were isolated from rust sori. Based on morphological data and DNA sequence analysis, the isolates were identified as three Lecanicillium species, viz. L. attenuatum, Lecanicillium sp. 1, Lecanicillium sp. 2, and V. epiphytum. The unidenified two species of Lecanicillium appear to be previously unknown taxa. Sixty-six isolates of miscellaneous hyphomycetes belonging to 6 species of 5 genera were obtained from pustules of rust fungi. On the basis of morphological and molecular analyses, the miscellaneous hyphomycetes growing on rusts were identified as Acrodontium crateriforme, Cladophialophora pucciniophila, Cladosporium cladosporioides, Phacellium vossianum, Ramularia coleosporii, and R. uredinicola.

  • PDF

Effects of Salt Concentration on Motility and Expression of Flagellin Genes in the Fish Pathogen Edwardsiella tarda (염 농도가 어류 병원체 Edwardsiella tarda의 운동성과 편모발현에 미치는 영향)

  • Yu, Jong-Earn;Park, Jun-Mo;Kang, Ho-Young
    • Journal of Life Science
    • /
    • v.21 no.10
    • /
    • pp.1487-1493
    • /
    • 2011
  • E. tarda, a fish pathogen, can survive in seawater under relatively high salt conditions as well as in fish under physiological salt conditions. Bacterial growth under different salt concentrations may influence the expression of genes involved in bacterial structure and physiology. The growth rate of E. tarda culture in high salt (3.5% NaCl) was similar to that in low salt (1.0% NaCl, physiological salt concentration). Interestingly, the strain moved much faster in low salt conditions than in high salt conditions. Electron microscopic observation demonstrated that the bacterial cells grown in high salt had less or no flagellation. Obvious flagellation was observed in the parental strain E. tarda CK41 grown in low-salt condition. Two putative genes coding flagellin were identified in the E. tarda genome sequences. The amino acid sequence comparison of each gene revealed 93% identities. A flagellin gene was PCR amplified and cloned into a cloning vector. Using an E. coli protein expression system, a part of flagellin protein was overexpressed. Using the purified protein, an anti-flagellin antibody was raised in the rabbit. Immunoblot analyses with flagellin specific antibody demonstrated that E. tarda CK41 expressed falgellin in low salt conditions, which is consistent with the results seen in motility assay and microscopic observation. This is the first report of salt regulated flagella expression in E. tarda.

Microbiological, Physicochemical, and Antioxidant Properties of Plain Yogurt and Soy Yogurt (플레인 요구르트와 소이 요구르트의 미생물학적, 물리화학적 및 항산화 활성)

  • Lim, Sung-Mee
    • Korean Journal of Microbiology
    • /
    • v.49 no.4
    • /
    • pp.403-414
    • /
    • 2013
  • This study evaluated the physicochemical and microbiological characteristics and antioxidant properties of yogurt samples fermented with lactic acid bacteria (LAB) obtained from pickled cabbage. API 50 CHL systems and 16S rRNA nucleotide sequence analyses revealed that the isolates were Lactobacillus casei PC05 and L. acidophilus PC16. Cell counts, titratable acidity, and viscosity of the yogurt samples fermented with L. acidophilus PC16 were significantly higher than those of the samples fermented with L. casei PC05 (P<0.05). The detected cell counts and physicochemical properties were significantly lower in plain yogurt than in soy yogurt (P<0.05). Yogurt samples fermented with L. acidophilus PC16 exhibited higher antioxidant activity, measured as ability to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and chelate ferrous ions, than those fermented with L. casei PC05. However, the ability to scavenge superoxide anions and superoxide dismutase (SOD) activity were significantly (P<0.05) higher in yogurt samples fermented with L. casei PC05 compared to those in samples fermented with L. acidophilus PC16. The antioxidant activity of soy yogurt was significantly (P<0.05) higher than that of plain yogurt. The antioxidant activity of the tested strains resulted in lipid peroxidation inhibition (in vitro), which may be related to the elimination of free radicals, chelating ability, and reducing power. There were no significant differences in the physicochemical properties and antioxidant activities of the yogurt samples during cold storage.

Characterization of a Cold Tolerance-related Gene, BrCSR, Derived from Brassica rapa (배추 유래 저온 저항성 관련 유전자, BrCSR의 특성 분석)

  • Yu, Jae-Gyeong;Park, Young-Doo
    • Horticultural Science & Technology
    • /
    • v.32 no.1
    • /
    • pp.91-99
    • /
    • 2014
  • The objective of this study is to identify cold-tolerance genes in Brassica rapa. In order to acheive this goal, we analyzed a KBGP-24K oligo chip data [BrEMD (B. rapa EST and Microarray Database)] using B. rapa ssp. pekinensis inbred line 'Chiifu' under cold stress condition ($4^{\circ}C$). Among 23,929 unigenes of B. rapa, 417 genes (1.7%) were primarily identified as cold responsive genes that were expressed over 5-fold higher than those of wild type control, and then a gene which has unknown function and has full length sequence was selected. It was named BrCSR (B. rapa Cold Stress Resistance). BrCSR was transformed using expression vector pSL101 to confirm whether BrCSR can enhance cold tolerance in tobacco plants. $T_1$ transgenic tobacco plants expressing BrCSR were selected by PCR and Southern hybridization analyses, and the function of BrCSR was characterized by expression level analysis and phenotype observation under cold stress condition. The expression level of BrCSR in transgenic tobacco plants increased up to about two folds in quantitative real-time RT-PCR assay and this was very similar to Northern blot hybridization analysis. Analysis of phenotypic characteristics clearly elucidated that transgenic tobaccos expressing BrCSR were more cold tolerant than wild type control under $4^{\circ}C$ treatment. Based on these results, we conclude that the over-expression of BrCSR might be closely related to the enhancement of cold tolerance.

CLINICAL STUDY OF CHILD AND ADOLESCENT PSYCHIATRIC OUTPATIENTS (소아 청소년 정신과 외래환자의 임상적 고찰)

  • Lee, He-Len;Hwang, Soon-Taeg
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
    • /
    • v.7 no.1
    • /
    • pp.14-22
    • /
    • 1996
  • This study aims to look at main problems of visiting the clinic, diagnoses and other related factors of outpatients in a private psychiatric clinic f3r children and adolescents located in Seoul. The analyses were based on the reports of 2,785 patients who were 18 years old and less, and visited the clinic during last 4 years. The results showed that the ratio of boys to girls was 2.7 to 1, and about 64% of the whole sample were 6 years old and less. Especially the percentage of patients aged 3 and less was the highest and that of schoolage and more was gradually reduced. The average number of siblings was 195 and the percentage of the first child in a family was the highest. Particularly, there were more boys in rase of one child families and more girls in case of families with 3 children and more. The chief problems were mainly language-deficit, hyperactivity, autistic behaviour, tic, aggressive behavior and academic problem. The higher frequency of diagnoses was in the order of parent-child problem, mental retardation, developmental language disorder, reactive attachment disorder, other emotional disorder, and pervasive developmental disorder. The more frequently used method fir treatments was in the sequence of psychotherapy, play therapy, parental counseling, occupational therapy and speech therapy. The results from this study were compared with those from other studies and discussed.

  • PDF

Diagnostic Performance of Diffusion-Weighted Steady-State Free Precession in Differential Diagnosis of Neoplastic and Benign Osteoporotic Vertebral Compression Fractures: Comparison to Diffusion-Weighted Echo-Planar Imaging

  • Shin, Jae Ho;Jeong, Soh Yong;Lim, Jung Hyun;Park, Jeongmi
    • Investigative Magnetic Resonance Imaging
    • /
    • v.21 no.3
    • /
    • pp.154-161
    • /
    • 2017
  • Purpose: To evaluate the diagnostic performance of diffusion-weighted steady-state free precession (DW-SSFP) in comparison to diffusion-weighted echo-planar imaging (DW-EPI) for differentiating the neoplastic and benign osteoporotic vertebral compression fractures. Materials and Methods: The subjects were 40 patients with recent vertebral compression fractures but no history of vertebroplasty, spine operation, or chemotherapy. They had received 3-Tesla (T) spine magnetic resonance imaging (MRI), including both DW-SSFP and DW-EPI sequences. The 40 patients included 20 with neoplastic vertebral fracture and 20 with benign osteoporotic vertebral fracture. In each fracture lesion, we obtained the signal intensity normalized by the signal intensity of normal bone marrow (SI norm) on DW-SSFP and the apparent diffusion coefficient (ADC) on DW-EPI. The correlation between the SI norm and the ADC in each lesion was analyzed using linear regression. The optimal cut-off values for the diagnosis of neoplastic fracture were determined in each sequence using Youden's J statistics and receiver operating characteristic curve analyses. Results: In the neoplastic fracture, the median SI norm on DW-SSFP was higher and the median ADC on DW-EPI was lower than the benign osteoporotic fracture (5.24 vs. 1.30, P = 0.032, and 0.86 vs. 1.48, P = 0.041, respectively). Inverse linear correlations were evident between SI norm and ADC in both neoplastic and benign osteoporotic fractures (r = -0.45 and -0.61, respectively). The optimal cut-off values for diagnosis of neoplastic fracture were SI norm of 3.0 in DW-SSFP with the sensitivity and specificity of 90.4% (95% confidence interval [CI]: 81.0-99.0) and 95.3% (95% CI: 90.0-100.0), respectively, and ADC of 1.3 in DW-EPI with the sensitivity and specificity of 90.5% (95% CI: 80.0-100.0) and 70.4% (95% CI: 60.0-80.0), respectively. Conclusion: In 3-T MRI, DW-SSFP has comparable sensitivity and specificity to DW-EPI in differentiating the neoplastic vertebral fracture from the benign osteoporotic vertebral fracture.

Availability of MADLDI-TOF MS for Identification of Gram Positive Bacilli Isolated from Blood Culture

  • Choi, Jin-Un;Kim, Sang-Ha;Hwang, Su-Jeong;Yu, Young-Bin;Kim, Sunghyun;Kim, Young-Kwon
    • Biomedical Science Letters
    • /
    • v.24 no.2
    • /
    • pp.108-115
    • /
    • 2018
  • In the present study, results of the identification of Gram-positive bacilli (GPB) were analyzed by using the MALDI-TOF MS technique to score each 2-year blood culture at a university hospital. In addition, 16S rRNA sequence analyses and MALDI-TOF MS results are compared to targeting strains that had been isolated two or more times within the same patient, to evaluate the usefulness of MALDI-TOF MS in GPB identification. According to the cut-off (${\geq}1.7$) criteria, there were 410 (57.5%) reliable strains and 303 (42.5%) non-identified strains among the GPB identification results of 713 strains, using a microflex MALDI Biotyper (Bruker Daltonik GmbH, Bremen, Germany). The isolation appeared most often in the following order: Corynebacterium striatum, Bacillus cereus, Bacillus subtilis, Paenibacillus urinalis, and Listeria monocytogenes. Nearly three-fourths, 66 out of 89 (74.2%) of the strains for Corynebacterium striatum; 44 out of 60 (73.3%) strains for Bacillus cereus; and all (25 out of 25, 100%) Listeria monocytogenes strains were identified by their high scores of 2.0 or higher. Most (293 strains out of 303) non-identified strains were strains isolated only once and not significant as infectious bacilli. A total of 43 out of 50 (86.0%) strains matched and were able to be identified based on the 16 rRNA sequencing comparison results of strains that were isolated twice or more within the same patient and significant as infection bacilli. Non-matching among 5 out of 7 strains was not identified, even with MALDI-TOF MS. In conclusion, GPB can be identified in blood cultures using MALDI-TOF MS. This can be done accurately with ease, rapidly, and at a low cost. It is also thought to be helpful in GPB diagnosis and treatment.