• Title/Summary/Keyword: 16S-rRNA

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Isolation and identification of 18 unrecorded prokaryotic species from the intestinal tracts of aquatic animals in Korea

  • Lee, Jae-Yun;Jeong, Yun-Seok;Kim, Pil Soo;Hyun, Dong-Wook;Bae, Jin-Woo
    • Journal of Species Research
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    • v.10 no.1
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    • pp.1-11
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    • 2021
  • Evidence highlighting the importance of gut microbiota in biodiversity conservation is growing; however, gut bacteria in South Korean wildlife have not been well identified. Using a culture-dependent isolation method, we identified the gut bacteria from Korean aquatic wildlife: the gazami crab (Portunus trituberculatus), Korean striped bitterling (Acheilognathus yamatsutae), oily bitterling (Acheilognathus koreensis), leopard mandarin fish (Siniperca scherzeri), Korean dark chub (Zacco koreanus), diving beetle (Cybister lewisianus), spotted steed (Abbottina springeri), and Korean spotted sleeper (Odontobutis obscura interrupta). We identified 18 strains previously unrecorded in South Korea by comparing 16S rRNA gene sequences of isolates against the EzBioCloud and National Institute of Biological Resources(NIBR) databases. The isolated strains belong to the phyla Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria. We also assessed for phylogenetic relatedness, Gram-stain reaction, colony and cell morphology, and biochemical characteristics. Basic information and 16S rRNA gene sequences of the isolates were registered in NIBR, and NIBR accession numbers are provided.

A new species and new record of Pacificincolidae (Bryozoa: Cheilostomata) from Korea

  • Min, Bum Sik;Chae, Hyun Sook;Yang, Ho Jin;Noh, Geon Woo;Lee, Dong Hee;Seo, Ji Eun
    • Journal of Species Research
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    • v.10 no.3
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    • pp.276-286
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    • 2021
  • We performed a taxonomic study on Korean Pacificincolidae (Bryozoa: Cheilostomata) with the materials collected from 39 localities(ports, intertidal and subtidal zones) in the coastal seas of South Korea from 1981 to 2020. Korean Mucronella perforata reported by Rho and Seo (1985) turned out to be not Pacificincola perforata, but Primavelans glabricollaris n. sp. based on morphological data. Pacificincola perforata, which was transferred from Mucronella perforata (Okada and Mawatari, 1937), is also added to the Korean fauna. This study is the first to use mitochondrial 16S rRNA gene sequences to study the family Paficincolidae. The NJ analysis also reconfirmed the validity of the genus Primavelans, established on the basis of the early astogeny by De Blauwe in 2006, with the mitochondrial 16S rRNA sequences. The inter-genetic distance among two species belong to the Pacificincolidae was in the range of 13.39±1.83 mm. Primavelans glabricollaris n. sp. is thus distinguished from Pacificincola perforata with both morphological and molecular data. A new species, Primavelans glabricollaris, and a new record, Pacificincola perforata from Korea are herein described in detail and illustrated by scanning electron microscopy.

Six newly recorded species of epilithic cyanobacteria isolated in Korea

  • Hye-Ryeung Wang;Ji-Ho Song;Nam-Ju Lee;Do-Hyun Kim;So-Won Kim;Ok-Min Lee
    • Journal of Species Research
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    • v.13 no.1
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    • pp.10-31
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    • 2024
  • In this study, 11 strains of epilithic cyanobacteria belonging to six unrecorded species in Korea were isolated from gravel submerged in freshwater of seven collection sites in Korea. The morphological characteristics of the six species isolated in this study were consistent with the type strain of each species, and the similarity of the 16S rRNA gene sequences with the type strain of each species were 98.8-100%. In the phylogenetic tree using the 16S rRNA gene sequences, the 11 strains of these six species formed the same cluster as the strains of each species. The habitat of each previously reported species is mainly the soil surface, but all Korean strains appeared from the gravel submerged in freshwater. As a result of the morphological, ecological, and molecular analyses, these six species of cyanobacteria were identified as Geminocystis papuanica, Allocoleopsis franciscana, Ancylothrix terrestris, Klisinema persicum, Scytolyngbya timoleontis, and Shackletoniella antarctica, which were added as newly recorded species in Korea.

Associations of physical activity with gut microbiota in pre-adolescent children

  • Santarossa, Sara;Sitarik, Alexandra R.;Johnson, Christine Cole;Li, Jia;Lynch, Susan V.;Ownby, Dennis R.;Ramirez, Alex;Yong, Germaine LM.;Cassidy-Bushrow, Andrea E.
    • Korean Journal of Exercise Nutrition
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    • v.25 no.4
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    • pp.24-37
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    • 2021
  • [Purpose] To determine whether physical activity (PA), primarily the recommended 60 minutes of moderate-to-vigorous PA, is associated with gut bacterial microbiota in 10-year-old children. [Methods] The Block Physical Activity Screener, which provides minutes/day PA variables, was used to determine whether the child met the PA recommendations. 16S rRNA sequencing was performed on stool samples from the children to profile the composition of their gut bacterial microbiota. Differences in alpha diversity metrics (richness, Pielou's evenness, and Faith's phylogenetic diversity) by PA were determined using linear regression, whereas beta diversity (unweighted and weighted UniFrac) relationships were assessed using PERMANOVA. Taxon relative abundance differentials were determined using DESeq2. [Results] The analytic sample included 321 children with both PA and 16S rRNA sequencing data (mean age [SD] =10.2 [0.8] years; 54.2% male; 62.9% African American), where 189 (58.9%) met the PA recommendations. After adjusting for covariates, meeting the PA recommendations as well as minutes/day PA variables were not significantly associated with gut richness, evenness, or diversity (p ≥ 0.19). However, meeting the PA recommendations (weighted UniFrac R2 = 0.014, p = 0.001) was significantly associated with distinct gut bacterial composition. These compositional differences were partly characterized by increased abundance of Megamonas and Anaerovorax as well as specific Christensenellaceae_R-7_group taxa in children with higher PA. [Conclusion] Children who met the recommendations of PA had altered gut microbiota compositions. Whether this translates to a reduced risk of obesity or associated metabolic diseases is still unclear.

Detection of Gram-negative Bacteria in Broad-range PCR Amplifying 16S rRNA Gene with Semi-nested Primers and Its Application in Market Milk (16S rRNA 유전자의 Semi-nested Primer를 이용한 Broad-range PCR에 의한 그람음성세균의 검출과 시유에서의 응용)

  • Choi, Suk-Ho;Choi, J.J.;Lee, S.B.
    • Journal of Animal Science and Technology
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    • v.47 no.3
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    • pp.465-474
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    • 2005
  • A two-step broad-range PCR method detecting gram-negative bacteria at the level as low as 2 CFU was developed by using primers of GNFI and GNRI and then semi-nested primer of GNF2 and GNRI. The nucleotide sequences of the primers were determined based on l6S rRNA gene. The DNA fragments of 1173 bp and 169 bp were amplified in one-step PCRs with primer sets of GNFI-GNRI and GNF2-GNRl, respectively, using template DNA from seven strains of gram-negative bacteria including Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas spp., and Acinetobacter baumaii but not from Achromobacter lyticus, Alca/igens faecalis, and five strains of gram-positive bacteria. DNA fragments of 180 bp were amplified from LTLT-pasteurized milk and UHf-pasteurized milk in the two-step PCR. The DNA fragments were amplified from LTLT-pasteurized milk which was added with Pseudomonas j/uorescens and subsequently heated at 65 $^{\circ}C$, 80 $^{\circ}C$, and 100 $^{\circ}C$ for 30 min but they were not amplified from the milk autoclaved at 121$^{\circ}C$ for 15 min. It was suggested in PCR that Pseudomonas fluorescens heated at 65 $^{\circ}C$ for 30 min in milk was more sensitive to DNase treatment than viable bacteria.

Biodiversity and Isolation of Gut Microbes from Digestive Organs of Harmonia axyridis (무당벌레 소화기관으로부터 장내세균의 분리 및 계통학적 다양성)

  • Kim, Ki-Kwang;Han, Song-Ih;Moon, Chung-Won;Yu, Yong-Man;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.47 no.1
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    • pp.66-73
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    • 2011
  • Bacterial density distributions of gut microbes in the digestive organs of Harmonia axyridis collected from three different sources (JK, CK, and CJ) were $6.0{\times}10^4$ CFU/gut under aerobic culture condition and $8.0{\times}10^6$ CFU/gut under anaerobic culture condition. Seven colony types were observed under aerobic condition and three types of similarity were detected under anaerobic condition. In total, 116 strains, including 34 strains under aerobic condition, were isolated from the digestive organs of H. axyridis. Based on the analysis of the 16S rRNA gene sequence, aerobic gut microbes were assigned to the Proteobacteria, Actinobacteria, Firmicutes, and Deinococcus-Thermus. A large number of isolates belonged to the genus Bacillus and Staphylococcus of the Firmicutes commonly found in H. axyridis from different sites. Anaerobic gut microbes were found to be similar according to colony morphological, phylogenetic analysis using ARDRA. Eighty-two anaerobic gut microbes were clustered into 17 different ARDRA types according to HaeIII. Representative anaerobic gut microbes in each ARDRA group were divided into five species of ${\gamma}$-Proteobacteria based on 16S rRNA gene sequence analysis; Hafnia alvei, Enterobacter ludwigii, Enterobacter kobei, Pseudomonas oryzihabitans and Pseudomonas koreensis. Phylogenetic analysis indicated that about 70% of the isolates belonged to ${\gamma}$-Proteobacteria, suggesting predominance of gut microbes.

Genome-based identification of strain KCOM 1265 isolated from subgingival plaque at the species level

  • Park, Soon-Nang;Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.45 no.2
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    • pp.70-75
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    • 2020
  • The aim of this study was to identify strain KCOM 1265 isolated from subgingival plaque at the species level by comparing 16S ribosomal RNA gene (16S rDNA) and genome sequences. The whole genome of strain KCOM 1265 was extracted using the phenol-chloroform extraction method. 16S rDNA was amplified using polymerase chain reaction and sequenced using the dideoxy chain termination method. Pairwise genome comparison was performed using average nucleotide identity (ANI) and genome-to-genome distance (GGD) analyses. The data showed that the percent similarity of 16S rDNA sequence of strain KCOM 1265 was 99.6% as compared with those of Fusobacterium polymorphum ATCC 10953T and Fusobacterium hwasookii KCOM 1249T. The ANI values of strain KCOM 1265 with F. polymorphum ATCC 10953T and F. hwasookii KCOM 1249T were 95.8% and 93.0%, respectively. The GGD values of strain KCOM 1265 with F. polymorphum ATCC 10953T and F. hwasookii KCOM 1249T were 63.9% and 49.6%, respectively. These results indicate that strain KCOM 1265 belongs to F. polymorphum.

Studies on the mycoplasmal pneumonia in slaughter pigs. 2. Isolation of mycoplasmas from lung tissues and identification of isolates by nested-PCR technique (도축돈의 마이코플라즈마성 폐렴에 관한 연구 2. 폐조직에서의 균분리와 nested-PCR방법에 의한 동정)

  • Lim, Young-Taek;Seok, Ho-Bong
    • Korean Journal of Veterinary Research
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    • v.42 no.2
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    • pp.225-229
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    • 2002
  • We report that mycoplasma organisms from lung tissues of slaughter pigs were identified to genes fragments with references use of nested-PCR technique(nPCR). Seven strains of mycoplasma species were isolated from 70 lung tissues. The organisms were detected by in vitro amplification of 16S rRNA and 23S rRNA genes. Nucleotide sequences of the spacer between 16S and 23S in the ribosomal RNA operons of mycoplasma were identified by the analysis of products from the nested PCR. Four common PCR primers, MhF1, MhF2 MhR1 and MhR2, were designed by analysis between these sequences by first amplified with F1, R1 and second with F2, R2, respectively. Specific amplification of the spacer region for reference strains of M. hyopneumoniae, M. hyorhinis, M. flocculare were confirmed by first round of PCR in which the traduced fragments of 690bp, 460bp, 630bp. But amplications of second round was changed to 240bp, 210bp, 230bp, respectively. Three different strains (M. hyopneumoniae:4, M. hyorhinis:2, M. flocculare:1) were detected by the nested-PCR technique. The results suggest that the detection of swine mycoplasma by n-PCR can be analyzed the nucleotide sequences between rRNA operons and homology study.

Microbial Communities of Activated Sludge in an Anaerobic/Aerobic Sequencing Batch Reactor using Slot Hybridization (Slot Hybridization을 이용한 연속 회분식 반응기내 미생물 분포 조사)

  • Jeon, Che Ok;Shin, Kum-Joo;Lee, Dae Sung;Suh, Pann-Ghill;Park, Jong Moon
    • Journal of Korean Society of Environmental Engineers
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    • v.22 no.5
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    • pp.939-947
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    • 2000
  • Enhanced biological phosphorus removal (EBPR) was performed in an anaerobic/aerobic sequencing batch reactor (SBR). Influent was a synthetic wastewater based on acetate as a carbon source. The sludge age and hydraulic retention time were kept at 10 days and 16 hrs, respectively, Phosphate release during the anaerobic period and phosphate uptake in aerobic period were increased gradually with time. and after about 200 days, steady-state operation could be achieved with complete removal of influent phosphate. Number distribution of microbial community in the sludge performing EBPR was investigated during the steady state operation. 17 rRNA targeted oligonucleotide probes were designed and slot hybridization technique was used to determine the number distribution of each microorganism. In the acetate fed SBR, rRNA belonging to the beta subclass of proteobacteria was the most dominant in total rRNA and rRNA matching to CTE probe was the second, rRNAs of Acinetobacter, Aeromonas and Pseudomonas, which are usually thought as phosphorus accumulating organisms in EBPR processes, constituted less than 10% of total rRNA. From this community analysis, it was inferred that microorganisms belong to the beta subclass of proteobacteia (BET) and CTE such as Rhodocyclus group were important in biological phosphorus removal. Therefore, the role of Acinetobacter, Aeromonas and Pseudomonas in the EBPR might have been overestimated.

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Antibiotic and Insecticidal Activities of Metabolites Derived From an Entomopathogenic Bacterium, Xenorhabdus nematophila, Against The Bean Bug, Riptortus clavatus (곤충병원세균(Xenorhabdus nematophila) 유래물질의 톱다리개미허리노린재(Riptortus clavatus) 장내세균 증식억제 및 살충효과)

  • Seo, Sam-Yeol;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.49 no.3
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    • pp.251-259
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    • 2010
  • A bacterial colony was isolated from the gut of the bean bug, Riptortus clavatus. From morphological and biochemical tests, the bacterial isolate showed the highest similarity to Staphylococcus succinus. DNA sequence of 16S rRNA gene of the bacterium supported the identification. Oral administration of penicillin G to adults of R. clavatus gave a dose-dependent mortality of adults of R. clavatus to adults along with significant decrease of the bacterial population in the gut. Similarly, three metabolites (benzylideneacetone, proline-tyrosine, and acetylated phenylalanine-glycine-valine) derived from an entomopathogenic bacterium, Xenorhabdus nematophila, also inhibited growth of the gut bacterial population and gave significant mortalities to R. clavatus. These results suggest that a gut bacterial population classified as Staphylococcus sp. is required for survival of R. clavatus and that the three bacterial metabolites had toxic effects on the bugs due to their antibacterial properties.