• Title/Summary/Keyword: Taxonomic characterization

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Novel Qnr Families as Conserved and Intrinsic Quinolone Resistance Determinants in Aeromonas spp.

  • Sang-Gyu Kim;Bo-Eun Kim;Jung Hun Lee;Dae-Wi Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.6
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    • pp.1276-1286
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    • 2024
  • The environment has been identified as an origin, reservoir, and transmission route of antibiotic resistance genes (ARGs). Among diverse environments, freshwater environments have been recognized as pivotal in the transmission of ARGs between opportunistic pathogens and autochthonous bacteria such as Aeromonas spp. In this study, five environmental strains of Aeromonas spp. exhibiting multidrug resistance (MDR) were selected for whole-genome sequencing to ascertain their taxonomic assignment at the species-level and to delineate their ARG repertoires. Analyses of their genomes revealed the presence of one protein almost identical to AhQnr (A. hydrophila Qnr protein) and four novel proteins similar to AhQnr. To scrutinize the classification and taxonomic distribution of these proteins, all Aeromonas genomes deposited in the NCBI RefSeq genome database (1,222 genomes) were investigated. This revealed that these Aeromonas Qnr (AQnr) proteins are conserved intrinsic resistance determinants of the genus, exhibiting species-specific diversity. Additionally, structure prediction and analysis of contribution to quinolone resistance by AQnr proteins of the isolates, confirmed their functionality as quinolone resistance determinants. Given the origin of mobile qnr genes from aquatic bacteria and the crucial role of Aeromonas spp. in ARG dissemination in aquatic environments, a thorough understanding and strict surveillance of AQnr families prior to the clinical emergence are imperative. In this study, using comparative genome analyses and functional characterization of AQnr proteins in the genus Aeromonas, novel Aeromonas ARGs requiring surveillance has suggested.

Characterization of microbial communities and soil organic carbon degradation associated with the depth and thawing effects on tundra soil in Alaska (Alaska 툰드라 토양의 깊이 및 해동 영향에 따른 미생물 군집과 토양 유기 탄소 분해 특성)

  • Park, Ha Ju;Kim, Dockyu;Park, Hyun;Lee, Bang Yong;Lee, Yoo Kyung
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.365-374
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    • 2016
  • In high-latitude regions, temperature has risen ($0.6^{\circ}C$ per decade) and this leads to the increase in microbial degradability against soil organic carbon (SOC). Furthermore, the decomposed SOC is converted into green-house gases ($CO_2$ and $CH_4$) and their release could further increase the rate of climate change. Thus, understanding the microbial diversity and their functions linked with SOC degradation in soil-thawing model is necessary. In this study, we divided tundra soil from Council, Alaska into two depth regions (30-40 cm and 50-60 cm of depth, designated as SPF and PF, respectively) and incubated that for 108 days at $0^{\circ}C$. A total of 111,804 reads were obtained through a pyrosequencing-based metagenomic study during the microcosm experiments, and 574-1,128 of bacterial operational taxonomic units (OTUs) and 30-57 of archaeal OTUs were observed. Taxonomic analysis showed that the distribution of bacterial taxa was significantly different between two samples. In detail, the relative abundance of phyla Actinobacteria and Firmicutes largely increased in SPF and PF soil, respectively, while phyla Crenarchaeota was increased in both soil samples. Weight measurement and gel permeation chromatography of the SOC extracts demonstrated that polymerization of humic acids, main component of SOC, occurred during the microcosm experiments. Taken together our results indicate that these bacterial and archaeal phyla could play a key function in SOC degradation and utilization in cold tundra soil.

Studies on Constituents of Higher Fungi of Korea (LXXI) -Application of Enzymes to Taxonomy of Ganoderma Species-

  • Kim, Byong-Kak;Kim, Jin-Sook;Choi, Kyun-Gae;Kim, Ha-Won;Choi, Eung-Chil
    • Korean Journal of Pharmacognosy
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    • v.24 no.2
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    • pp.116-123
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    • 1993
  • The genus Ganoderma is typical wood-rotting fungi and its fruiting body has been used as an important herb in oriental medicine. Recent research discovered antitumor components from Ganoderma lncidum. Various Ganoderma species are being cultivated in Korea. However, taxonomic system of the genus Ganoderma has been based mainly on the macromorphology of fruiting bodies and the ultrastructural characteristics of basidiospores. Since there are similar characteristics in Ganoderma mycelia grown on the same artificial media, it is suggested that the compatibility of the fungi by di-mon mating be used as an aid to determine the identity of species in addition to the conventional characterization. In this study, we examined physiological and genetical properties such as growth temperature, pH, compatibility and enzyme or protein patterns of laccase, esterase and cellular proteins of G. lucidum RZ, G. tsugae and Ganoderma species cultivated in Korea by electrophoresis for characterization of the isolates. We found that compatibility test and isozyme patterns of laccase and esterase of the mycelia could be used for the differentiation of the isolates. These results showed that Ganoderma species cultivated in Korea is genetically similar to G. lucidum but physiologically closer to G. tsugae than to G. lucidum.

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Production and Characterization of Cholesterol Oxidase from Streptomyces sp. No.4 (방선균으로부터 Cholesterol Oxidase의 생산 및 특성)

  • 김현수;고희선
    • KSBB Journal
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    • v.14 no.2
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    • pp.174-180
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    • 1999
  • An actinomycetes strain No.4 which produce the cholesterol oxidase(EC 1.1.3.6), was isolated from soil and identified as Streptomyces sp. based on taxonomic studies. The conditions of cholesterol oxidase production and enzymatic properties were investigated. The optimum composition of medium for production of the enzyme was 1% soluble starch, 2% corn steep liquor, 0.1% $KH_2PO_4$, 0.1% $NaNO_3$ and 0.05% $MgSO_4$ (pH 7.0). The optimum pH and temperature of the cholesterol oxidase were pH 6.0~7.5 and $37^{\circ}C$, respectively. The enzyme was stable in the range of pH 6.0~9.0. The isoelectric point determined by multichambered electrofocusing unit was in the range of pH 6.0~6.5.

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Morphological and Molecular Characterization of Thamnocalamus falconeri Hook f. ex. Munro

  • Tiwari, Chandrakant;Bakshi, Meena;Nautiyal, Subhash
    • Journal of Forest and Environmental Science
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    • v.31 no.3
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    • pp.214-224
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    • 2015
  • The economy of India and so also of many Asian countries depends on bamboos and their uses are not only in domestic items but also in rural housing and raw materials to several industries and germplasm characterization is an important link between the conservation and utilization of plant genetic resources. Classical taxonomic studies of the bamboos are based on floral morphology and growth habit, which can cause problems in identification due to erratic flowering coupled with different biotic agencies and environmental factors. Identification and genetic relationships among accessions of Thamnocalamus falconeri were investigated using morphology and random amplified polymorphic DNAs (RAPD) technique. Analysis started by using 51 vegetative characters and forty two 10-mer primers that allowed us to distinguish different genotypes hailing from different eco- zones of Garhwal Himalayas (India). The selected primers (12) were used for identification and for establishing a profiling system to estimate genetic diversity. A total of 79.33% polymorphism was estimated by using 12 selected primers. The genetic similar analysis was conducted based on binary digits i.e. presence (1) or absence (0) of bands, which revealed a wide range of variability among the species whereas genetic relatedness was quite high based on vegetative characters. Cluster analysis clearly showed two major clusters for both of the markers viz. morphology and RAPD belonging to 10 accessions of T. falconeri. Two major clusters were further divided into minor clusters. Cluster based on RAPD marker showed grouping of accessions of closed locality whereas analogy was reported for vegetative traits. The RAPD technique has the potential for use in species identification and genetic relationships studies of bamboo for breeding program.

Isolation, Production, and Characterization of Protease from Bacillus subtilis IB No. 11

  • Lee, Min-Hyang;Lee, Kang-Moon;Choi, Yong-Jin;Baek, Yeon-Soo
    • Journal of Animal Science and Technology
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    • v.51 no.6
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    • pp.527-536
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    • 2009
  • A potent protein degrading bacterium was isolated from soil samples of different environments. Polyphasic taxonomic studies and phylogenetic 16S rRNA sequence analyses led to identify the isolate IB No. 11 as a strain of Bacillus subtilis. The isolated strain was recognized to produce protease constitutively, and the maximum production (1.64 units/ml) was attained in a shake flask culture when the isolate was grown at $40^{\circ}C$, for 32 h in basal medium supplemented with starch (0.25%) and gelatin (1.25%) as sole carbon and nitrogen source, respectively. The optimum pH and temperature for the protease activity were determined to be pH 7.0 and $50^{\circ}C$, respectively. $Ca^{2+}$ and $Mn^{2+}$ enhanced remarkably the protease activity but neither showed positive effect on the protease's thermal stability. In addition, it was observed that the protease was fairly stable in the pH range of 6.5-8.0 and at temperatures below $50^{\circ}C$, and it could be a good candidate for an animal feed additive. The inhibition profile of the protease by various inhibitors indicated that the enzyme is a member of serine-proteases. A combination of UV irradiation and NTG mutagenesis allowed to develop a protease hyper-producing mutant strain coded as IB No. 11-4. This mutant strain produced approximately 3.23-fold higher protease activity (6.74 units/mg) than the parent strain IB No. 11 when grown at $40^{\circ}C$ for 32h in the production medium. The protease production profile of the selected mutants was also confirmed by the zymography analysis.

Phylogenetic Diversity of Acidophilic Sporoactinobacteria Isolated from Various Soils

  • Cho, Sung-Heun;Han, Ji-Hye;Seong, Chi-Nam;Kim, Seung-Bum
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.600-606
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    • 2006
  • Spore forming actinobacteria (sporoactinobacteria) isolated from soils with an acidic pH in Pinus thunbergii forests and coal mine waste were subjected to taxonomic characterization. For the isolation of acidophilic actinobacteria, acidified starch casein agar (pH adjusted to 4-5) was used. The numbers of actinobacteria growing in acidic media were between $3.2{\times}10^4$ and $8.0{\times}10^6$ CFU/g soil. Forty three acidophilic actinobacterial strains were isolated and their 16S rDNA sequences were determined. The isolates were divided into eight distinctive phylogenetic clusters within the variation encompassed by the family Streptomycetaceae. Four clusters among them were assigned to the genus Streptacidiphilus, whereas the remaining four were assigned to Streptomyces. The clusters belonging to either Streptomyces or Streptacidiphilus did not form a monophyletic clade. The growth pH profiles indicated that the representative isolates grew best between pH 5 and 6. It is evident from this study that acidity has played a critical role in the differentiation of the family Streptomycetaceae, and also that different mechanisms might have resulted in the evolution of two groups, Streptacidiphilus (strict acidophiles) and neutrotolerant acidophilic Streptomyces. The effect of geographic separation was clearly seen among the Streptacidiphilus isolates, which may be a key factor in speciation of the genus.

Application of Molecular Biology to Rumen Microbes -Review-

  • Kobayashi, Y.;Onodera, R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.1
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    • pp.77-83
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    • 1999
  • Molecular biological techniques that recently developed, have made it possible to realize some of new attempts in the research field of rumen microbiology. Those are 1) cloning of genes from rumen microorganisms mainly in E. coli, 2) transformation of rumen bacteria and 3) ecological analysis with nonculturing methods. Most of the cloned genes are for polysaccharidase enzymes such as endoglucanase, xylanase, amylase, chitinase and others, and the cloning rendered gene structural analyses by sequencing and also characterization of the translated products through easier purification. Electrotransformation of Butyrivibrio fibrisolvens and Prevotella ruminicola have been made toward the direction for obtaining more fibrolytic, acid-tolerant, depoisoning or essential amino acids-producing rumen bacterium. These primarily required stable and efficient gene transfer systems. Some vectors, constructed from native plasmids of rumen bacteria, are now available for successful gene introduction and expression in those rumen bacterial species. Probing and PCR-based methodologies have also been developed for detecting specific bacterial species and even strains. These are much due to accumulation of rRNA gene sequences of rumen microbes in databases. Although optimized analytical conditions are essential to reliable and reproducible estimation of the targeted microbes, the methods permit long term storage of frozen samples, providing us ease in analytical work as compared with a traditional method based on culturing. Moreover, the methods seem to be promissing for obtaining taxonomic and evolutionary information on all the rumen microbes, whether they are culturable or not.

Characterization of Algal Community of Yongdam Reservoir and Identification of Ecological Factors Inducing the Changes in Community Composition (용담호 조류군집의 시공간적 분포와 조류발생 요인분석)

  • Kim, Hyun-su;Jeong, Il-hwan
    • Journal of Soil and Groundwater Environment
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    • v.20 no.7
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    • pp.121-134
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    • 2015
  • Spatial and temporal changes in algal population in Yongdam reservoir and ecological factors that induced the changes in the size and composition of algal population were investigated by monthly sampling at ten locations in the reservoir. Nutritional state of the reservoir was identified to be phosphorus-limited with nitrogen to phosphorus (N : P) ratio much greater than 17 in most samples. Algal population was dominated by three taxonomic groups, diatoms, chlorophytes and cyanobacteria. Although explosive algal growth was not observed in the summer, algal population showed transition with time of the dominant algal type from diatoms in the winter to cyanobacteria in the summer. Chlorophyta was not the dominant group in the reservoir although they maintained relatively stable number of cells in the reservoir and showed increase in population from March to May. The application of statistical methods revealed that the factors inducing changes in cell number of each group were water temperature for diatoms and cyanobacteria and phosphorus concentration for chlorophyte. Fluctuation of cyanobacterial population was mainly observed near the inlet of tributaries while diatoms showed higher variation inside the reservoir.

Molecular Characterization of Nippostrongylus brasiliensis (Nematoda: Heligmosomatidae) from Mus musculus in India

  • Chaudhary, Anshu;Goswami, Urvashi;Singh, Hridaya Shanker
    • Parasites, Hosts and Diseases
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    • v.54 no.6
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    • pp.743-750
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    • 2016
  • Mus musculus (Rodentia: Muridae) has generally been infected with a rodent hookworm Nippostrongylus brasiliensis. In this report, we present morphological and molecular identification of N. brasiliensis by light and scanning electron microscopy and PCR amplification of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and the protein sequences encoded by cox1 gene, respectively. Despite the use of N. brasiliensis in many biochemistry studies from India, their taxonomic identification was not fully understood, especially at the species level, and no molecular data is available in GenBank from India. Sequence analysis of cox1 gene in this study revealed that the present specimen showed close identity with the same species available in GenBank, confirming that the species is N. brasiliensis. This study represents the first record of molecular identification of N. brasiliensis from India and the protein structure to better understand the comparative phylogenetic characteristics.