• Title/Summary/Keyword: 16S rRNA analysis

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Isolation, Characterization and Whole-Genome Analysis of Paenibacillus andongensis sp.nov. from Korean Soil

  • Yong Guan;Zhun Li;Yoon-Ho Kang;Mi-Kyung Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.753-759
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    • 2023
  • The genus Paenibacillus contains a variety of biologically active compounds that have potential applications in a range of fields, including medicine, agriculture, and livestock, playing an important role in the health and economy of society. Our study focused on the bacterium SS4T (KCTC 43402T = GDMCC 1.3498T), which was characterized using a polyphasic taxonomic approach. This strain was analyzed using antiSMASH, BAGEL4, and PRISM to predict the secondary metabolites. Lassopeptide clusters were found using all three analysis methods, with the possibility of secretion. Additionally, PRISM found three biosynthetic gene clusters (BGC) and predicted the structure of the product. Genome analysis indicated that glucoamylase is present in SS4T. 16S rRNA sequence analysis showed that strain SS4T most closely resembled Paenibacillus marchantiophytorum DSM 29850T (98.22%), Paenibacillus nebraskensis JJ-59T (98.19%), and Paenibacillus aceris KCTC 13870T (98.08%). Analysis of the 16S rRNA gene sequences and Type Strain Genome Server (TYGS) analysis revealed that SS4T belongs to the genus Paenibacillus based on the results of the phylogenetic analysis. As a result of the matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/MS) results, SS4T was determined to belong to the genus Paenibacillus. Comparing P. marchantiophytorum DSM 29850T with average nucleotide identity (ANI 78.97%) and digital DNA-DNA hybridization (dDDH 23%) revealed values that were all less than the threshold for bacterial species differentiation. The results of this study suggest that strain SS4T can be classified as a Paenibacillus andongensis species and is a novel member of the genus Paenibacillus.

Reduction of Hydrogen Sulphide in Chicken Manure by Immobilized Sulphur Oxidising Bacteria Isolated from Hot Spring

  • Hidayat, M.Y.;Saud, H.M.;Samsudin, A.A.
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.116-124
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    • 2019
  • The rapid development of the poultry industry has led to the production of large amounts of manure, which produce substances like hydrogen sulfide ($H_2S$) that contribute to odor pollution. $H_2S$ is a highly undesirable gas component and its removal from the environment is therefore necessary. Sulfur-oxidizing bacteria (SOB) are widely known to remove contaminating $H_2S$ due to their ability to oxidize reduced sulfur compounds. In this study, three potential SOB (designated AH18, AH25, and AH28) that were previously isolated from a hot spring in Malaysia were identified by 16S rRNA gene analysis. Laboratory-scale biological deodorization experiments were conducted to test the performance of the three isolates-in the form of pure or mixed cultures, with the cells immobilized onto alginate as a carrier-in reducing the $H_2S$ from chicken manure. On the basis of 16S rRNA phylogenetic analysis, isolate AH18 was identified as Pseudomonas sp., whereas isolates AH25 and AH28 were identified as Achromobacter sp. The most active deodorizing isolate was AH18, with an $H_2S$ reduction rate of 74.7% (p < 0.05). Meanwhile, the reduction rates for isolates AH25 and AH28 were 54.2% and 60.8% (p > 0.05), respectively. However, the $H_2S$ removal performance was enhanced in the mixed culture, with a reduction rate of 81.9% (p < 0.05). In conclusion, the three potential SOB isolates were capable of reducing the $H_2S$ from chicken manure in the form of a pure culture immobilized on alginate, and the reduction performance was enhanced in the mixed culture.

Molecular Systematics of Tephritidae (Insecta : Diptera): Testing Phylogenetic Position of Korean Acidiella spp. (Trypetini) Using Mitochondrial 16S rDNA Sequences

  • Han, Ho-Yeon;Ro, Kyung-Eui
    • Animal cells and systems
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    • v.6 no.1
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    • pp.13-18
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    • 2002
  • Phylogenetic relationships of Korean Acidiella species were tested using mitochondrial 16S ribosomal RNA gene sequences. We used 16 published sequences as outgroup, and 10 new sequences for nine Korean Acidiella species as ingroup. The number of aligned sites was 1,281 bp, but 1,135 bp were used for the analysis after excluding sites with missing data or gaps. Among these 1,135 sites, 464 sites were variable and 340 were informative for parsimony analysis. Phylogenetic information was extracted from this data set using neighbor-joining, maximum likelihood and maximum parsimony methods and compared to a morphology-based phylogenetic hypothesis. Our molecular data suggest that: (1) the tribe Trypetini appears to be monophyletic even when the nine additional Acidiella species are added to our previous phylogenetic analysis; (2) all the Korean Acidiella species belong to the Trypeta group, but the genus Acidiella is not supported as monophyletic; (3) the close relationship of A. circumvaga, A. issikii, and A. sapporensis is supported; (4) the close relationship of A. pachypogon and two additional new Acidiella species is strongly supported; and (5) the possible presence of two or more cryptic species among the specimens previously identified as A. obscuripennis is suggested. Sequence data from the mitochondrial 16S rDNA allowed us to better understand the systematic status of Korean Acidiella species. They indicated that the current concept about the genus Acidiella is insufficient and needs to be refined further. This study also showed a few interesting relationships, that had not been recognized by morphological study alone. Based on this study, we were able to plan further experiments to analyze relationships within the Trypeta Group.

Association of miR-193b Down-regulation and miR-196a up-Regulation with Clinicopathological Features and Prognosis in Gastric Cancer

  • Mu, Yong-Ping;Tang, Song;Sun, Wen-Jie;Gao, Wei-Min;Wang, Mao;Su, Xiu-Lan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8893-8900
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    • 2014
  • Dysregulated expression of microRNAs (miRNAs) has been shown to be closely associated with tumor development, progression, and carcinogenesis. However, their clinical implications for gastric cancer remain elusive. To investigate the hypothesis that genome-wide alternations of miRNAs differentiate gastric cancer tissues from those matched adjacent non-tumor tissues (ANTTs), miRNA arrays were employed to examine miRNA expression profiles for the 5-pair discovery stage, and the quantitative real-time polymerase chain reaction (qRTPCR) was applied to validate candidate miRNAs for 48-pair validation stage. Furthermore, the relationship between altered miRNA and clinicopathological features and prognosis of gastric cancer was explored. Among a total of 1,146 miRNAs analyzed, 16 miRNAs were found to be significantly different expressed in tissues from gastric cancer compared to ANTTs (p<0.05). qRT-PCR further confirmed the variation in expression of miR-193b and miR-196a in the validation stage. Down-expression of miR-193b was significantly correlated with Lauren type, differentiation, UICC stage, invasion, and metastasis of gastric cancer (p<0.05), while over-expression of miR-196a was significantly associated with poor differentiation (p=0.022). Moreover, binary logistic regression analysis demonstrated that the UICC stage was a significant risk factor for down-expression of miR-193b (adjusted OR=8.69; 95%CI=1.06-56.91; p=0.043). Additionally, Kaplan-Meier survival curves indicated that patients with a high fold-change of down-regulated miR-193b had a significantly shorter survival time (n=19; median survival=29 months) compared to patients with a low fold-change of down-regulated miR-193b (n=29; median survival=54 months) (p=0.001). Overall survival time of patients with a low fold-change of up-regulated miR-196a (n=27; median survival=52 months) was significantly longer than that of patients with a high fold-change of up-regulated miR-196a (n=21; median survival=46 months) (p=0.003). Hence, miR-193b and miR-196a may be applied as novel and promising prognostic markers in gastric cancer.

Natural occurrence of Mycobacterium as an endosymbiont of Acanthamoeba isolated from a contact lens storage case

  • Yu, Hak-Sun;Jeong, Hae-Jin;Hong, Yeon-Chul;Seol, Seong-Yong;Chung, Dong-Il;Kong, Hyun-Hee
    • Parasites, Hosts and Diseases
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    • v.45 no.1 s.141
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    • pp.11-18
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    • 2007
  • Recent in vitro studies have revealed that a certain Mycobacterium can survive and multiply within free-living amoebae. It is believed that protozoans function as host cells for the intracellular replication and evasion of Mycobacterium spp. under harmful conditions. In this study, we describe the isolation and characterization of a bacterium naturally observed within an amoeba isolate acquired from a contact lens storage case. The bacterium multi-plied within Acanthamoeba, but exerted no cytopathic effects on the amoeba during a 6-year amoebic culture. Trasnmission electron microscopy showed that the bacteria were randomly distributed within the cytoplasm of trophozoites and cysts of Acanthamoeba. On the basis of the results of 18S rRNA gene analysis, the amoeba was identified as A. lugdunensis. A 16S rRNA gene analysis placed this bacterium within the genus Mycobacterium. The bacterium evidenced positive reactivity for acid-fast and fluorescent acid-fast stains. The bacterium was capable of growth on the Middlebrook 7H11-Mycobacterium-specific agar. The identification and characterization of bacterial endosymbionts of free-living protozoa bears significant implications for our understanding of the ecology and the identification of other atypical mycobacterial pathogens.

Effects of Field-Grown Genetically Modified Zoysia Grass on Bacterial Community Structure

  • Lee, Yong-Eok;Yang, Sang-Hwan;Bae, Tae-Woong;Kang, Hong-Gyu;Lim, Pyung-Ok;Lee, Hyo-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.333-340
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    • 2011
  • Herbicide-tolerant Zoysia grass has been previously developed through Agrobacterium-mediated transformation. We investigated the effects of genetically modified (GM) Zoysia grass and the associated herbicide application on bacterial community structure by using culture-independent approaches. To assess the possible horizontal gene transfer (HGT) of transgenic DNA to soil microorganisms, total soil DNAs were amplified by PCR with two primer sets for the bar and hpt genes, which were introduced into the GM Zoysia grass by a callus-type transformation. The transgenic genes were not detected from the total genomic DNAs extracted from 1.5 g of each rhizosphere soils of GM and non-GM Zoysia grasses. The structures and diversities of the bacterial communities in rhizosphere soils of GM and non-GM Zoysia grasses were investigated by constructing 16S rDNA clone libraries. Classifier, provided in the RDP II, assigned 100 clones in the 16S rRNA gene sequences library into 11 bacterial phyla. The most abundant phyla in both clone libraries were Acidobacteria and Proteobacteria. The bacterial diversity of the GM clone library was lower than that of the non- GM library. The former contained four phyla, whereas the latter had seven phyla. Phylogenetic trees were constructed to confirm these results. Phylogenetic analyses of the two clone libraries revealed considerable difference from each other. The significance of difference between clone libraries was examined with LIBSHUFF statistics. LIBSHUFF analysis revealed that the two clone libraries differed significantly (P<0.025), suggesting alterations in the composition of the microbial community associated with GM Zoysia grass.

Investigation of Bacterial Diversity in Membrane Bioreactor and Conventional Activated Sludge Processes from Petroleum Refineries Using Phylogenetic and Statistical Approaches

  • Silva, Cynthia;Jesus, Ederson C.;Torres, Ana P. R.;Sousa, Maira P.;Santiago, Vania M. J.;Oliveira, Valeria M.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.3
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    • pp.447-459
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    • 2010
  • Bacterial diversity of two distinct wastewater treatment systems, conventional activated sludge (CAS) and membrane bioreactor (MBR), of petroleum refineries were investigated through 16S rRNA gene libraries. Sequencing and phylogenetic analysis showed that the bacterial community composition of sludge samples was distinct between the two wastewater treatment systems. MBR clones belonged predominantly to Class Betaproteobacteria, represented mainly by genera Thiobacillus and Thauera, whereas CAS clones were mostly related to Class Alphaproteobacteria, represented by uncultured bacteria related to Order Parvularculales. Richness estimators ACE and Chao revealed that the diversity observed in both libraries at the species level is an underestimate of the total bacterial diversity present in the environment and further sampling would yield an increased observed diversity. Shannon and Simpson diversity indices were different between the libraries and revealed greater bacterial diversity for the MBR library, considering an evolutionary distance of 0.03. LIBSHUFF analyses revealed that MBR and CAS communities were significantly different at the 95% confidence level ($P{\leq}0.05$) for distances $0{\leq}D{\leq}0.20$. This work described, qualitatively and quantitatively, the structure of bacterial communities in industrial-scale MBR and CAS processes of the wastewater treatment system from petroleum refineries and demonstrated clearly differentiated communities responsible for the stable performance of wastewater treatment plants.

Freeze-drying feces reduces illumina-derived artefacts on 16S rRNA-based microbial community analysis (Illumina를 이용한16S rRNA 기반 미생물생태분석에서 분변의 동결건조에 의한 인공적인 시퀀스 생성 감소효과)

  • Kim, Jungman;Unno, Tatsuya
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.299-304
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    • 2016
  • When used for amplicon sequencing, Illumina platforms produce more than hundreds of sequence artefacts, which affects operational taxonomic units based analyses such as differential abundance and network analyses. Nevertheless it has become a major tool for fecal microbial community analysis. In addition, results from sequence-based fecal microbial community analysis vary depending on conditions of samples (i.e., freshness, time of storage and quantity). We investigated if freeze-drying samples could improve quality of sequence data. Our results showed reduced number of possible artefacts while maintaining overall microbial community structure. Therefore, freeze-drying feces prior to DNA extraction is recommended for Illumina-based microbial community analysis.

Novel Sporichthyaceae Bacterium Strain K-07 Skin Barrier, Moisturizing and Anti-inflammatory Activity (신규 Sporichthyaceae Bacterium Strain K-07 배양액의 피부장벽, 보습 및 항염증 활성)

  • Lee, Dong-Geol;Kim, Minji;Kang, Seunghyun;Kim, Youn-Joon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.2
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    • pp.137-147
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    • 2017
  • The human skin is an ecosystem that provides habitat to various microorganisms. These comprise the skin microbiome and provide numerous benefits in addition to maintaining a symbiotic relation with the host. Various metabolites generated by the skin microbiome exert beneficial effects such as strengthening the skin barrier, and anti-aging and anti-inflammatory functions. In this study, we isolated a novel bacterium, designated Sporichthyacae strain K-07, from the human skin. Analysis of 16S rRNA gene sequences showed that the newly found bacterium shares 93.4% homology with the genus Sporichthya, thus corroborating the discovery of a novel genus. We further analyzed the effect of the novel strain in vitro, by treating HaCaT cells with bacterial metabolite products. Treatment resulted in changes in the mRNA expression levels of filaggrin, claudin1, claudin4, SMase, CERS3, HAS3, aquaporin3, IL-6, TNF-${\alpha}$, TSLP, and TARC. Specifically, the levels of filaggrin, claudin1, claudin4, SMase, CERS3, HAS3, and aquaporin3 were higher in strain K-07 metabolite product-treated cells than in control cells. These results showed that metabolite products of the novel strain K-07 enhanced the skin barrier and exert anti-inflammatory effects. Therefore, these metabolite products could be potentially used for treatment of skin conditions.

Bacterial Diversity of Culturable Isolates from Seawater and a Marine Coral, Plexauridae sp., near Mun-Sum, Cheju-Island

  • Lee, Jung-Hyun;Shin, Hyun-Hee;Lee, Deuk-Soo;Kwon, Kae-Kyung;Kim, Sang-Jin;Lee, Hong-Kum
    • Journal of Microbiology
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    • v.37 no.4
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    • pp.193-199
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    • 1999
  • Fifty-eight strains showing different colony morphological characteristics on various media were isolated from marine coral (Plexauridae sp.) and ambient seawater near Mun-Sum, Cheju-Island in 1998. Bacterial diversity was studies by phylogenetic analysis of the partial 16S rRNA gene sequences. All isolates representing the bacterial domain included affiliates of the high G+C (59%) and los G+C (3%) subdivision of Gram positive bacteria, and the alpha (33%) and gamma (5%) subdivision of the Proteobacteria. The 16S rDNA sequence similarity of the isolates was in the 88.3 to 100% range (average, 95.6%) to reported sequence data. In the comparison of the isolates from marine coarl and ambient seawater, more diverse groups belonging to ${\alpha}$-Proteobacteria were preferentially obtained from seawater.

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