• Title/Summary/Keyword: 16S-rRNA

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Investigation of Microbial Contamination of Public Bath in Jongno-gu, Seoul (서울특별시 종로구 대중목욕탕의 수질 중 미생물 오염도 조사 연구)

  • Kim, Mi-Soon;Lee, Young-Min;Kim, Seong-Keun;Seo, Ji-Hyun;Ji, Kyung-Hee;Oh, Ji-Yoon;Ko, Ki-Dong;Ko, Gwang-Pyo
    • Journal of Environmental Health Sciences
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    • v.35 no.3
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    • pp.162-168
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    • 2009
  • The aim of this study is to investigate microbial sanitary condition of public baths in Seoul, Korea. A total of 28 water samples were collected from 14 different public baths and sudatoriums. The prevalence of fecal indicator microorganisms such as total coliform, fecal coliform, and Escherichia coli was characterized. In addition, bacteria in water was membrane filtered by 0.45um nitrocellulose membrane, and the filter was analyzed by both cultivation and PCR amplification of partial 16S rRNA gene. The levels of chlorine were measured for each of water samples. More than 40% of 14 collected water samples, the concentrations of total coliform bacteria exceeded the water quality for bath water guideline. There was no significant correlation between chlorine residue and the presence of total coliform. Various microorganisms including pathogenic microorganisms were identified from cultivation and subsequent analysis of 16s rRNA gene sequences. Our results suggest that appropriate hygiene practice and continuous monitoring is needed for reducing health risk associated with public bathhouses.

Metagenomic SMRT Sequencing-Based Exploration of Novel Lignocellulose-Degrading Capability in Wood Detritus from Torreya nucifera in Bija Forest on Jeju Island

  • Oh, Han Na;Lee, Tae Kwon;Park, Jae Wan;No, Jee Hyun;Kim, Dockyu;Sul, Woo Jun
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1670-1680
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    • 2017
  • Lignocellulose, composed mostly of cellulose, hemicellulose, and lignin generated through secondary growth of woody plant, is considered as promising resources for biofuel. In order to use lignocellulose as a biofuel, biodegradation besides high-cost chemical treatments were applied, but knowledge on the decomposition of lignocellulose occurring in a natural environment is insufficient. We analyzed the 16S rRNA gene and metagenome to understand how the lignocellulose is decomposed naturally in decayed Torreya nucifera (L) of Bija forest (Bijarim) in Gotjawal, an ecologically distinct environment. A total of 464,360 reads were obtained from 16S rRNA gene sequencing, representing diverse phyla; Proteobacteria (51%), Bacteroidetes (11%) and Actinobacteria (10%). The metagenome analysis using single molecules real-time sequencing revealed that the assembled contigs determined originated from Proteobacteria (58%) and Actinobacteria (10.3%). Carbohydrate Active enZYmes (CAZy)- and Protein families (Pfam)-based analysis showed that Proteobacteria was involved in degrading whole lignocellulose, and Actinobacteria played a role only in a part of hemicellulose degradation. Combining these results, it suggested that Proteobacteria and Actinobacteria had selective biodegradation potential for different lignocellulose substrates. Thus, it is considered that understanding of the systemic microbial degradation pathways may be a useful strategy for recycle of lignocellulosic biomass, and the microbial enzymes in Bija forest can be useful natural resources in industrial processes.

Isolation and characterization of a new Methanoculleus bourgensis strain KOR-2 from the rumen of Holstein steers

  • Battumur, Urantulkhuur;Lee, Manhee;Bae, Gui Sek;Kim, Chang-Hyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.2
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    • pp.241-248
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    • 2019
  • Objective: To isolate and identify new methanogens from the rumen of Holstein steers in Korea. Methods: Representative rumen contents were obtained from three ruminally cannulated Holstein steers ($793{\pm}8kg$). Pre-reduced media were used for the growth and isolation of methanogens. Optimum growth temperature, pH, and sodium chloride (NaCl) concentration as well as substrate utilization and antibiotic tolerance were investigated to determine the physiological characteristics of the isolated strain. Furthermore, the isolate was microscopically studied for its morphology. Polymerase chain reaction of 16S rRNA and mcrA gene-based amplicons was used for identification. Results: One strain designated as KOR-2 was isolated and found to be a non-motile irregular coccus with a diameter of 0.2 to $0.5{\mu}m$. KOR-2 utilized $H_2+CO_2$ and formate but was unable to metabolize acetate, methanol, trimethylamine, 2-propanol, and isobutanol for growth and methane production. The optimum temperature and pH for the growth of KOR-2 were $38^{\circ}C$ and 6.8 to 7.0, respectively, while the optimum NaCl concentration essential for KOR-2 growth was 1.0% (w/v). KOR-2 tolerated ampicillin, penicillin G, kanamycin, spectromycin, and tetracycline. In contrast, the cell growth was inhibited by chloramphenicol. Phylogenetic analysis of 16S rRNA and mcrA genes revealed the relatedness between KOR-2 and Methanoculleus bourgensis. Conclusion: Based on the physiological and phylogenetic characteristics, KOR-2 was thought to be a new strain within the genus Methanoculleus and named Methanoculleus bourgensis KOR-2.

Amazonocrinis thailandica sp. nov. (Nostocales, Cyanobacteria), a novel species of the previously monotypic Amazonocrinis genus from Thailand

  • Tawong, Wittaya;Pongcharoen, Pongsanat;Pongpadung, Piyawat;Ponza, Supat;Saijuntha, Weerachai
    • ALGAE
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    • v.37 no.1
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    • pp.1-14
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    • 2022
  • Cyanobacteria are distributed worldwide, and many new cyanobacterial species are discovered in tropical region. The Nostoc-like genus Amazonocrinis has been separated from the genus Nostoc based on polyphasic methods. However, species diversity within this genus remains poorly understood systematically because only one species (Amazonocrinis nigriterrae) has been described. In this study, two novel strains (NUACC02 and NUACC03) were isolated from moist rice field soil in Thailand. These two strains were characterized using a polyphasic approach, based on morphology, 16S rRNA phylogenetic analysis, internal transcribed spacer secondary structure and ecology. Phylogenetic analyses based on 16S rRNA gene sequences confirmed that the two novel strains formed a monophyletic clade related to the genus Amazonocrinis and were distant from the type species A. nigriterrae. The 16S rRNA gene sequence similarity (<98.1%) between novel strains and all other closely related taxa including the Amazonocrinis members exceeded the cutoff for species delimitation in bacteriology, reinforcing the presence of a new Amazonocrinis species. Furthermore, the novel strains possessed unique phenotypic characteristics such as the presence of the sheath, necridia-like cells, larger cell dimension and akinete cell arrangement in long-chains and the singularity of D1-D1', Box-B, V2, and V3 secondary structures that distinguished them from other Amazonocrinis members. Considering all the results, we described our two strains as Amazonocrinis thailandica sp. nov. in accordance with the International Code of Nomenclature for Algae, Fungi and Plants.

Microbial Contamination according to the Numbers of Mask Worn in the Community

  • Eun Ju Lee;Heechul Park;Min-A Je;Songhee Jung;Gahee Myoung;Su Bin Jo;Hyun Min Hwang;Ryeong Si;Hyunwoo Jin;Kyung-Eun Lee;Jungho Kim
    • Biomedical Science Letters
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    • v.28 no.4
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    • pp.317-321
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    • 2022
  • Due to COVID-19 pandemic, wearing face masks is obligatory to prevent respiratory virus transmissions in the community. However, there are few studies of the desirable number of wearing a face mask, and how to store them for reuse. Therefore, in this study, a survey was conducted among 208 healthy adults, and 27 kf-94 masks worn for 1, 2, and 3 days were collected. To estimate the risk of bacterial contamination, we analyzed the extent of bacterial contamination of the BHI medium and 16S rRNA gene sequencing. With an increase in the number of days of using the mask, the degree of bacterial contamination of the used mask gradually increased. As a result of 16S rRNA PCR performed for strain identification, Staphylococcus, known as a pathogenic bacterium, was identified the most. In conclusion, we found that wearing a cotton KF mask provides an optimal environment for microbes, which are related to the skin and respiratory system, to thrive. Therefore, it is also important to reduce the risk of bacterial infection of the face mask with appropriate sterilization methods.

A report of 20 unrecorded bacterial species isolated from the coastal area of Korean islands in 2022

  • Hyerim Cho;Yeonjung Lim;Sumin Kim;Hyunyoung Jo;Mirae Kim;Jang-Cheon Cho
    • Journal of Species Research
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    • v.12 no.2
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    • pp.165-173
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    • 2023
  • Bacterial communities inhabiting islands play a vital role in the functioning and formation of a unique, isolated ecosystem. Nevertheless, there has been a lack of systematic research on the indigenous microbiological resources of the islands in Korea. To excavate microbial resources for further studies on the metabolism and biotechnological potential, a standard dilution plating was applied to coastal seawater samples collected from islands along the west coast of the Korean Peninsula, including Deokjeokdo, Baengnyeongdo, and Daebudo in 2022. A total of 2,007 bacterial strains were isolated from the samples as single colonies and identified using 16S rRNA gene sequence analyses. A total of 20 strains, with ≥98.7% 16S rRNA gene sequence similarity to bacterial species having validly published names but not reported in Korea, were designated as unrecorded bacterial species in Korea. The unrecorded bacterial strains were phylogenetically diverse and belonged to four phyla, five classes, 12 orders, 17 families, and 18 genera. The unreported species were assigned to Algimonas, Amylibacter, Notoacmeibacter, Roseibium, and Terasakiella of the class Alphaproteobacteria; Alteromonas, Congregibacter, Marinagarivorans, Marinicella, Oceanospirillum, Psychromonas, Thalassotalea, Umboniibacter, and Vibrio of the class Gammaproteobacteria; Lutibacter and Owenweeksia of the class Flavobacteriia; Paenibacillus of the class Bacilli; and Pelagicoccus of the class Opitutae. The taxonomic characteristics of the unreported species, including morphology, biochemistry, and phylogenetic position are provided in detail.

Exploring the Feasibility of 16S rRNA Short Amplicon Sequencing-Based Microbiota Analysis for Microbiological Safety Assessment of Raw Oyster

  • Jaeeun Kim;Byoung Sik Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.9
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    • pp.1162-1169
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    • 2023
  • 16S rRNA short amplicon sequencing-based microbiota profiling has been thought of and suggested as a feasible method to assess food safety. However, even if a comprehensive microbial information can be obtained by microbiota profiling, it would not be necessarily sufficient for all circumstances. To prove this, the feasibility of the most widely used V3-V4 amplicon sequencing method for food safety assessment was examined here. We designed a pathogen (Vibrio parahaemolyticus) contamination and/or V. parahaemolyticus-specific phage treatment model of raw oysters under improper storage temperature and monitored their microbial structure changes. The samples stored at refrigerator temperature (negative control, NC) and those that were stored at room temperature without any treatment (no treatment, NT) were included as control groups. The profiling results revealed that no statistical difference exists between the NT group and the pathogen spiked- and/or phage treated-groups even when the bacterial composition was compared at the possible lowest-rank taxa, family/genus level. In the beta-diversity analysis, all the samples except the NC group formed one distinct cluster. Notably, the samples with pathogen and/or phage addition did not form each cluster even though the enumerated number of V. parahaemolyticus in those samples were extremely different. These discrepant results indicate that the feasibility of 16S rRNA short amplicon sequencing should not be overgeneralized in microbiological safety assessment of food samples, such as raw oyster.

Discovery of 18 previously unrecorded bacterial species in the coastal areas surrounding Korean islands in 2023

  • Yeonjung Lim;Hyeonuk Sa;Minjeong Kim;Minseok Kim;Jisoo Han;Hyerim Cho;Jang-Cheon Cho
    • Journal of Species Research
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    • v.13 no.3
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    • pp.318-325
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    • 2024
  • Bacterial communities residing on islands have a significant impact on the functioning and establishment of a unique isolated ecosystem. Notwithstanding, systematic research on the indigenous microbial resources of domestic islands has been lacking. In order to understand the biodiversity and potential bioresources, we conducted sampling in 2023 from coastal waters from various islands off the west coast of the Korean Peninsula, including Baengnyeongdo, Daebudo, Deokjeokdo, Jangbongdo, Yeonpyeongdo Islands, along with Somaemuldo Island along the south coast. The coastal seawater samples were used to unearth microbial resources through the standard dilution plating. In total, approximately 1,600 bacterial strains were isolated from the samples as single colonies and identified using 16S rRNA gene sequence analyses. Eighteen strains, exhibiting ≥98.7% 16S rRNA gene sequence similarity to bacterial species with validly published names but not previously reported in Korea, were categorized as unrecorded bacterial species in Korea. These unrecorded bacterial strains displayed phylogenetic diversity, representing three phyla, four classes, 9 orders, 13 families, and 18 genera. The unrecorded species were assigned to the classes Alphaproteobacteria (Aliiroseovarius, Kiloniella, Maritalea, Palleronia, and Roseobacter), Gammaproteobacteria (Aliamphritea, Aliivibrio, Enterovibrio, Francisella, Leucothrix, Pseudoalteromonas, Psychrobium, Shewanella, and Vibrio), Flavobacteriia (Aquimarina, Pseudofulvibacter, and Tenacibaculum), and Verrucomicrobiae (Roseibacillus). This study presents comprehensive descriptions of the taxonomic attributes of these unrecorded species, covering morphology, biochemistry, and phylogenetic position.

'Bring to Lab' of 19 Novel Species Among 60 Isolates Retrieved from a Freshwater Pond

  • Song, Jae-Ho;Yang, Seung-Jo;Cho, Jang-Cheon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.168-175
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    • 2007
  • We report here on the cultivation of numerous novel bacterial species from a eutrophic freshwater pond. A total of 60 strains, 15 strains per each culture medium, were obtained from the surface of a eutrophic freshwater pond by employing a conventional dilution-plating method with four different kinds of culture media, including R2A, 1/10R2A, PCA, and 1/10PCA. Among the 60 strains isolated, 27 strains showed less than 97% 16S rRNA gene sequence similarities to validly published species, and thus they are considered to comprise 19 novel species. Of the 27 strains assigned to the novel species, the majority of the strains (20 strains) were affiliated with the Alphaproteobacteria and Betaproteobacteria. The remaining 7 strains were affiliated with the Gammaproteobacteria, Firmicutes, Actinobacteria, and Deinococci. Because we have isolated 19 novel species from a usual freshwater pond using a conventional culturing technique, our results suggest that an unexplored ecosystem, even if it looks like a common ecosystem found elsewhere, harbors diverse unidentified microbes, which will be definitely further characterized.

Phylogenetic Diversity of Dominant Bacterial and Archaeal Communities in Plant-Microbial Fuel Cells Using Rice Plants

  • Ahn, Jae-Hyung;Jeong, Woo-Suk;Choi, Min-Young;Kim, Byung-Yong;Song, Jaekyeong;Weon, Hang-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1707-1718
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    • 2014
  • In this study, the phylogenetic diversities of bacterial and archaeal communities in a plant-microbial fuel cell (P-MFC) were investigated together with the environmental parameters, affecting its performance by using rice as a model plant. The beneficial effect of the plant appeared only during a certain period of the rice-growing season, at which point the maximum power density was approximately 3-fold higher with rice plants. The temperature, electrical conductivity (EC), and pH in the cathodic and anodic compartments changed considerably during the rice-growing season, and a higher temperature, reduced difference in pH between the cathodic and anodic compartments, and higher EC were advantageous to the performance of the P-MFC. A 16S rRNA pyrosequencing analysis showed that the 16S rRNAs of Deltaproteobacteria and those of Gammaproteobacteria were enriched on the anodes and the cathodes, respectively, when the electrical circuit was connected. At the species level, the operational taxonomic units (OTUs) related to Rhizobiales, Geobacter, Myxococcus, Deferrisoma, and Desulfobulbus were enriched on the anodes, while an OTU related to Acidiferrobacter thiooxydans occupied the highest proportion on the cathodes and occurred only when the circuit was connected. Furthermore, the connection of the electrical circuit decreased the abundance of 16S rRNAs of acetotrophic methanogens and increased that of hydrogenotrophic methanogens. The control of these physicochemical and microbiological factors is expected to be able to improve the performance of P-MFCs.