• Title/Summary/Keyword: 16S rRNA analysis

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Analysis of Total Bacteria, Enteric Members of γ-proteobacteria and Microbial Communities in Seawater as Indirect Indicators for Quantifying Biofouling

  • Lee, Jin-Wook;Kim, Sung-Min;Jung, Ji-Yeon;Oh, Byung-Soo;Kim, In S.;Hong, Soon-Kang
    • Environmental Engineering Research
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    • v.14 no.1
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    • pp.19-25
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    • 2009
  • In this study, total bacteria, enteric members of the $\gamma$-proteobacteria, and microbial communities in seawater were analyzed as indirect indicators for quantifying biofouling. Biomass in seawater can significantly affect feed water pretreatment and membrane biofouling of reverse osmosis desalination processes. The purpose of this paper is to investigate microbiological quantity and quality of seawater at the potential intake of a desalination plant. For this analysis, the total direct cell count (TDC) using 4'-6-diamidino-2-phenylindole (DAPI)-staining and DNA-based real-time PCR were used to quantify the total bacteria and relative content of enteric members of $\gamma$-proteobacteria in seawater, respectively. In addition, microbial communities were examined using 16S rRNA gene cloning and bacterial isolation to identify the most abundant bacteria for a further biofouling study. The experimental results of this study identified about $10^6$ cells/mL of (total) bacteria, $10^5$ 16S rRNA gene copies/mL of enteric $\gamma$-proteobacteria, and the presence of more than 20 groups of bacteria.

Biokinetics of Protein Degrading Clostridium cadaveris and Clostridium sporogenes in Batch and Continuous Mode of Operations

  • Koo, Taewoan;Jannat, Md Abu Hanifa;Hwang, Seokhwan
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.533-539
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    • 2020
  • A quantitative real-time polymerase chain reaction (QPCR) was applied to estimate biokinetic coefficients of Clostridium cadaveris and Clostridium sporogenes, which utilize protein as carbon source. Experimental data on changes in peptone concentration and 16S rRNA gene copy numbers of C. cadaveris and C. sporogenes were fitted to model. The fourth-order Runge-Kutta approximation with non-linear least squares analysis was employed to solve the ordinary differential equations to estimate biokinetic coefficients. The maximum specific growth rate (μmax), half-saturation concentration (Ks), growth yield (Y), and decay coefficient (Kd) of C. cadaveris and C.sporogenes were 0.73 ± 0.05 and 1.35 ± 0.32 h-1, 6.07 ± 1.52 and 5.67 ± 1.53 g/l, 2.25 ± 0.75 × 1010 and 7.92 ± 3.71 × 109 copies/g, 0.002 ± 0.003 and 0.002 ± 0.001 h-1, respectively. The theoretical specific growth rate of C. sporogenes always exceeded that of C. cadaveris at peptone concentration higher than 3.62 g/l. When the influent peptone concentration was 5.0 g/l, the concentration of C.cadaveris gradually decreased to the steady value of 2.9 × 1010 copies/ml at 4 h Hydraulic retention time (HRT), which indicates a 67.1% reduction of the initial population, but the wash out occurred at HRTs of 1.9 and 3.2 h. The 16S rRNA gene copy numbers of C. sporogenes gradually decreased to steady values ranging from 1.1 × 1010 to 2.9 × 1010 copies/ml. C. sporogenes species was predicted to wash out at an HRT of 1.6 h.

Analysis of the Bacterial Community in Ojingeo-jeotgal and Selection of Bacillus Species Inhibiting the Growth of Food Pathogens (오징어젓갈 Bacteria 군집분석 및 식중독균 생육저해 Bacillus 균주 선발)

  • Kim, Hye-Rim;Han, Seulhwa;Lee, Bitnara;Jeong, Do-Won;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.41 no.4
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    • pp.462-468
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    • 2013
  • Jeotgal is a generic term given to the high-salt-fermented seafood of Korea. This study aimed at developing an overview of the bacterial community present in Ojingeo-jeotgal, a highly consumed type of jeotgal, which is made with squid. Bacteria were isolated and purified from two samples on six different kinds of media and identified by 16S rRNA gene sequence analysis. Among the 121 total isolates, the most dominant genus was Bacillus, followed by coagulase-negative staphylococci (CNS) and lactic acid bacteria (LAB). CNS were detected in both samples, but LAB were observed in only a single sample. Six strains of Bacillus species inhibiting the growth of food pathogens, Staphylococcus aureus and Vibrio parahaemolyticus, were selected from the 121 isolates. These were found to inhibit the growth of both pathogens in addition to displaying proteolytic activities on media containing 6% NaCl and 2% skim milk.

Antibacterial Activity of Streptomyces Strains Isolated from Different Regions of Jordan

  • Hala Khyami-Horani;Amal Al-Aboudi;Musa Abu Zarga;Monther Sadder;Halima Othman
    • Microbiology and Biotechnology Letters
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    • v.51 no.4
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    • pp.474-483
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    • 2023
  • Members of the genus Streptomyces produce more than 70% of antibiotics. The rise in antibiotic resistance globally enhanced the search for novel species with the ability to produce new bioactive compounds. This study was initiated to investigate different regions in Jordan for previously uncultured and rare Streptomyces species capable of producing novel antimicrobial compounds especially active against bacteria resistant to antibiotics. A total of 191 Streptomyces strains were isolated from 26 soil samples collected from different geographic regions in Jordan. Isolates were characterized based on colony and cellular morphology as well as using 16S rRNA gene sequencing. These isolates were screened for their ability to produce antibiotics by the perpendicular-cross streak method, and then tested by well diffusion method against tested pathogens. Fifty-four isolates showed potential to produce antimicrobial products especially active against resistant bacteria, 20.1% of the isolates showed inhibitory effect against Staphylococcus aureus, 16.9% against clinical MSSA strains, and 18.0% against MRSA: whereas only 4.2% against Esherichia coli, 3.2% against Klebsiella pneumonia, 2.7% against Pseudomonas aeruginosa, and 10.0% against clinical Candida albicans. Three isolates were selected for further identification due to their antibacterial activity against S. aureus, MRSA, and MSSA. These isolates were identified as follows; Streptomyces aburaviensis DSa3, Streptomyces alboniger SAb7 and Streptomyces misionensis ZAb2, based on cultural, biochemical characteristics and molecular analysis of the 16S rRNA.

Gluconacetobacter persimmonis sp. nov., Isolated from Korean Traditional Persimmon Vinegar

  • Yeo, Soo-Hwan;Lee, Oh-Seuk;Lee, In-Seon;Kim, Hyun-Soo;Yu, Tae-Shick;Jeong, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.276-283
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    • 2004
  • Screening was performed to isolate cellulose-producing microorganisms from the Korean traditional fermented persimmon vinegar. The resulting strain, KJ $145^{T}$, was then taxonomically investigated by phenotypic characterization, particularly chemotaxonomic, and by phylogenetic inference based on a 16S rDNA sequence analysis including other related taxa. Strain KJ $145^{T}$ was found to grow rapidly and form pale white colonies with smooth to rough surfaces on a GYC agar. Strain KJ $145^T$ also produced acetate from ethanol, and was tolerable to 10% ethanol in SM medium. In a static culture, a thick cellulose pellicle was produced, and in GYC broth, the strain grew at temperatures ranging from 28 to $40^\circ{C}$ with an optimum pH of 4.0. The genomic DNA G+C content of strain KJ $145^T$ was 61.9 mol%, and the predominant ubiquinone was Q 10 as the major quinone and Q9 as the minor quinone. The major cellular fatty acids were $C_{16:0}$ and the sum in feature 7 ($C_{18:1}$ w9c, w12t and/or w7c). A 16S rRNA-targeted oligonucleotide probe specific for strain KJ $145^T$was constructed, and the phylogenetic position of the new species was derived from a 16S rDNA-based tree. When comparing the 16S rDNA nucleotide sequences, strain KJ $145^T$ was found to be most closely related to G. hansenii LMG $1527^T$ (99.2%), although KJ $145^T$ was still distinct from G. hansenii LMG $l527^T$ and G. xylinus LMG $1515^T$ in certain phenotypic characteristics. Therefore, on the basis of 16S rDNA sequences and taxonomic characteristics, it is proposed that strain KJ $145^T$ should be placed in the genus Gluconacetobacter as a new species, Gluconacetobacter persimmonis sp. nov., under the type-strain KJ $145^T$ (=KCTC =$10175BP^T$=KCCM=$10354^T$).

Protease Properties of Protease-Producing Bacteria Isolated from the Digestive Tract of Octopus vulgaris (Octopus vulgaris의 장관으로부터 분리한 단백질 분해효소 생성 균주와 생성된 효소의 특성)

  • Liu, Qing;Ren, Pei;Piao, Meizi;Yang, Ji-Young
    • Journal of Life Science
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    • v.23 no.12
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    • pp.1486-1494
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    • 2013
  • A high protease-producing strain was isolated and identified from the digestive tract of octopus vulgaris by detecting a hydrolysis circle of protease and its activity. The strain was identified by morphology observation, biochemical experiments, and 16S rRNA sequence analysis. The protease obtained from the strain was purified by a three-step process involving ammonium sulfate precipitation, carboxy methyl-cellulose (CM-52) cation-exchange chromatography, and DEAE-Sephadex A50 anion-exchange chromatography. The properties of protease were characterized as well. The strain Bacillus sp. QDV-3, which produced the highest activity of protease, was isolated. On the basis of the phenotypic and biochemical characterization and 16S rRNA gene-sequencing studies, the isolate was identified as follows: domain: Bacteria; phylum: Firmicutes; class: Bacilli; order: Bacillales; family: Bacillaceae; and genus: Bacillus. The isolate was shown to have a 99.2% similarity with Bacillus flexus. A high active protease designated as QDV-E, with a molecular weight of 61.6 kDa, was obtained. The enzyme was found to be active in the pH range of 9.0-9.5 and its optimum temperature was $40^{\circ}C$. The protease activity retained more than 96% at the temperature of $50^{\circ}C$ for 60 min. Phenylmethylsulfonyl fluoride (PMSF) inhibited the enzyme activity, thus confirming that this protease isolated from Bacillus sp. QDV-3 is an alkaline serine protease. Metal ions, $Mn^{2+}$ and $Mg^{2+}$, were determined to enhance the protease activity, whereas $Ba^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ were found to inactivate the enzyme.

Isolation and characterization of Bacillus amyloliquefaciens TK3 inhibiting causative bacteria of atrophic rhinitis and fowl typhoid (돼지위축성비염과 가금티푸스 병원균을 저해하는 Bacillus amyloliquefaciens TK3의 분리 및 특성 조사)

  • Jung, Taeck-Kyung;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.177-180
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    • 2015
  • For prevention of atrophic rhinitis of swine by Bordetella bronchiseptica and fowl typhoid by Salmonella gallinarum, bacterial strains showing antimicrobial activity against those pathogenic bacteria were isolated from various samples collected at animal farms. Among 372 bacterial isolates strain TK3 showed the highest antibacterial activity against both pathogens, and was identified as Bacillus amyloliquefaciens by 16S rRNA gene sequence analysis. B. amyloliquefaciens TK3 could inhibit growth of both pathogens by secretion of antibacterial compounds such as siderophore, rhamnolipid and antimicrobial peptide. Production radius of siderophore on Chrome azurol S agar plate by strain TK3 was 0.53 cm after 14 days of incubation, and concentration of siderophore in King's B medium was 1.06 mmol/ml. It also secreted 82.4 mg/L of rhamnolipid, and antimicrobial peptide that completely inhibited growth of both pathogens at concentration of $30{\mu}l/ml$ in LB medium.

A report of six unrecorded radiation-resistant bacterial species isolated from soil in Korea in 2018

  • Maeng, Soohyun;Sathiyaraj, Srinivasan;Subramani, Gayathri;Kim, Ju-Young;Jang, Jun Hwee;Kang, Myung-Suk;Lee, Ki-Eun;Lee, Eun-young;Kim, Myung Kyum
    • Journal of Species Research
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    • v.7 no.3
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    • pp.222-230
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    • 2018
  • Six bacterial strains 18JY42-3, 18SH, 18JY76-11, 17J11-11, 18JY14-14, and 18JY15-11 assigned to the phylum Proteobacteria, Firmicutes, and Actinobacteria were isolated from soil samples in Korea. The Cohnella species, strain 18JY42-3 was Gram-stain-positive, short rod-shaped and beige-colored. The Methylobacterium species, strains 18SH and 18JY76-11 were Gram-stain-negative, short rod-shaped and pink-colored. The Microterricola species, strain 17J11-11 was Gram-stain-positive, short rod-shaped and yellow-colored. The Paenarthrobacter species, strains 18JY14-14 and 18JY15-11 were Gram-stain-positive, short rod-shaped and white-colored. Phylogenetic analysis based on 16S rRNA gene sequence showed that strains 18JY42-3, 18SH, 18JY76-11, 17J11-11, 18JY14-14, and 18JY15-11 were most closely related Cohnella rhizosphaerae (MH497628; 98.8%), Methylobacterium goesingense (MH497632; 99.1%), Methylobacterium populi (MH497635; 99.9%), Microterricolagilva (MH504108; 98.4%), Paenarthrobacter nicotinovorans (MH497641; 100%), and Paenarthrobacter nitroguajacolicus (MH497646; 99.2%), respectively. All the six unrecorded strains showed resistance to UV radiation. This is the first report of these six species in Korea.

Analysis of Potential Toxigenicity and Phylogeny using Target Genes in Aphanizomenon flos-aquae (Cyanophyceae) strains isolated from the Nakdong River (낙동강에서 분리된 Aphanizomenon flos-aquae (Cyanophyceae) 균주의 목표 유전자를 이용한 잠재적 독소 생성능 및 계통학적 분석)

  • Ryu, Hui-Seong;An, Sung-Min;Lim, Chang-Kun;Shin, Ra-Young;Park, Jong-Guen;Lee, Jung-Ho
    • Korean Journal of Ecology and Environment
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    • v.50 no.1
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    • pp.137-147
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    • 2017
  • The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.

Species Diversity of Betaproteobacteria in the Sumunmulbengdui Wetland Area of Jeju Island and Distribution of Novel Taxa (제주도 숨은물벵뒤 습지 서식 Betaproteobacteria의 종다양성 및 신분류군 분포)

  • Shin, Young-Min;Kim, Tae-Ui;Choi, Ah-Young;Chun, Jee-Sun;Lee, Sang-Hoon;Kim, Ha-Neul;Yi, Ha-Na;Jo, Jae-Hyung;Cho, Jang-Cheon;Jahng, Kwang-Yeop;Kim, Kyu-Joong;Joh, Ki-Seong;Chun, Jong-Sik;Lee, Hyune-Hwan;Kim, Seung-Bum
    • Korean Journal of Environmental Biology
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    • v.29 no.3
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    • pp.154-161
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    • 2011
  • The species diversity of Betaproteobacteria in the Sumunmulbengdui Wetland Area of Jeju Island was studied using culture based techniques, and candidates for novel taxa were screened. Twenty two novel bacterial strains belonging to Betaproteobacteria were isolated, which could be assigned to 16 genera of 4 families, namely Burkholderiaceae (3 strains), Comamonadaceae (8 strains), Oxalobacteraceae (5 strains), Neisseriaceae (5 strains), and an unassigned group belonging to Burkholderiales (1 strain) based 16S rRNA gene sequences. The genus Chromobacterium contained three candidates of novel species, and each of the genera Burkholderia, Comamonas, Pelomonas and Herbaspirillum contained two candidates respectively. Through the analysis of membrane fatty acid profiles and physiological properties using API 20NE as well as morphological and cultural properties, each of the isolates was found to form potentially novel species. Brief description of 22 potential candidates for new species or subspecies is given accordingly.