• Title/Summary/Keyword: Bacterial 16S rRNA sequencing

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Isolation and Characterization of Flavobacterium johnsoniae from Farmed Rainbow Trout Oncorhynchus mykiss

  • Suebsing, Rungkarn;Kim, Jeong-Ho
    • Fisheries and Aquatic Sciences
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    • v.15 no.1
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    • pp.83-89
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    • 2012
  • Flavobacterium johnsoniae was isolated from farmed rainbow trout Oncorhynchus mykiss in Korea, and its biochemical and molecular characterization was determined. Yellow-pigmented bacterial colonies were isolated from 18 of 64 fish samples (28.1%) on trypticase soy agar plates, and their biochemical profiles were characterized by API 20E and API 20NE test kits. F. johnsoniae was identified by biochemical phenotyping of factors including rapid gliding motility, Gram-negative condition, oxidase- and catalase-positive status, Congo red absorption, nitrate reduction, ${\beta}$-galactosidase production, acid production from glucose, and gelatin and casein hydrolysis. PCR and subsequent sequencing of 16S rRNA confirmed that the yellow-pigmented colonies were most similar to F. johnsoniae. The alignment analysis of 16S rRNA sequences also showed that all 18 rainbow trout isolates had highly similar homologies (97-99% identity). One isolate was selected and named FjRt09. This isolate showed 98% homology with previously reported F. johnsoniae isolates, and in phylogenetic analysis was more closely grouped with F. johnsoniae than with F. psychrophilum, F. columnare, or F. branchiophilum. This is the first report on the occurrence and biochemical characterization of F. johnsoniae isolated from rainbow trout in Korea.

Community Analysis of Nitrite-Oxidizing Bacteria in Lab-Scale Wastewater Treatment System (폐수처리장치에서의 아질산염 산화 세균 군집 분석)

  • Jeong, Soon-Jae;Lee, Sang-Ill;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.29-36
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    • 2008
  • Nitrogen is one of the major pollutants that should be removed by wastewater treatment systems. Biological nitrogen removal (BNR) is a key technology in advanced wastewater treatment systems operated by bacterial populations. Nitrification is the first step of microbiological processes in BNR system. Ammonia is oxidized to nitrite by ammonia-oxidizing bacteria (AOB) and then nitrite is subsequently oxidized to nitrate by nitrite-oxidizing bacteria (NOB). The diversity of NOB in nitrification reactors of 3 BNR systems, Edited biological aerated filter system, Nutrient removal laboratory system, and the Rumination type sequencing batch reactor system, was investigated by terminal restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA genes. Cluster analysis of T-RF profiles showed that communities of Nitrobacter group in each system were different depending upon the process of systems. However, the clusters of Nitrospira group were divided by the habitat of aqueous and solid samples.

Evaluation, Characterization and Molecular Analysis of Cellulolytic Bacteria from Soil in Peshawar, Pakistan

  • Ikram, Hira;Khan, Hamid Ali;Ali, Hina;Liu, Yanhui;Kiran, Jawairia;Ullah, Amin;Ahmad, Yaseen;Sardar, Sadia;Gul, Alia
    • Microbiology and Biotechnology Letters
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    • v.50 no.2
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    • pp.245-254
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    • 2022
  • Cellulases are a group of biocatalyst enzymes that are capable of degrading cellulosic biomass present in the natural environment and produced by a large number of microorganisms, including bacteria and fungi, etc. In the current study, we isolated, screened and characterized cellulase-producing bacteria from soil. Three cellulose-degrading species were isolated based on clear zone using Congo red stain on carboxymethyl cellulose (CMC) agar plates. These bacterial isolates, named as HB2, HS5 and HS9, were subsequently characterized by morphological and biochemical tests as well as 16S rRNA gene sequencing. Based on 16S rRNA analysis, the bacterial isolates were identified as Bacillus cerus, Bacillus subtilis and Bacillus stratosphericus. Moreover, for maximum cellulase production, different growth parameters were optimized. Maximum optical density for growth was also noted at pH 7.0 for 48 h for all three isolates. Optical density was high for all three isolates using meat extract as a nitrogen source for 48 h. The pH profile of all three strains was quite similar but the maximum enzyme activity was observed at pH 7.0. Maximum cellulase production by all three bacterial isolates was noted when using lactose as a carbon rather than nitrogen and peptone. Further studies are needed for identification of new isolates in this region having maximum cellulolytic activity. Our findings indicate that this enzyme has various potential industrial applications.

Bacterial Community Dynamics during Composting of Food Wastes (음식물 쓰레기 퇴비화 과정에 따른 세균군집 구조의 변화)

  • Shin, Ji-Hye;Lee, Jin-Woo;Nam, Ji-Hyun;Park, Se-Yong;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.148-154
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    • 2009
  • Composting is a biological process converting solid organic waste into valuable materials such as fertilizer. The change of bacterial populations in a composting reactor of food waste was investigated for 2 months. Based on shifts in temperature profile, the composting process could be divided into the first phase ($2^{\circ}C\sim55^{\circ}C$), the second phase ($55^{\circ}C\sim97^{\circ}C$), and the third phase ($50^{\circ}C\sim89^{\circ}C$). The number of total bacteria was $1.66\times10^{11}$ cell/g, $0.29\times10^{11}$ cell/g, and $0.28\times10^{11}$ cell/g in the first, second, and third stages, respectively. The proportions of thermophiles increased from 33% to 89% in the second stage. T-RFLP analysis and nucleotide sequencing of 16S rRNA gene demonstrated that the change of bacterial community structure was coupled with shifts in composting stages. The structure of bacterial community in the ultra-thermophilic second stage reflected that of seeding starter. The major decomposers driving the ultra-thermophilic composting were identified as phylotypes related to Bacillus and Pseudomonas.

High Level of Bacterial Diversity and Novel Taxa in Continental Shelf Sediment

  • Hong, Jin-Kyung;Cho, Jae-Chang
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.771-779
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    • 2012
  • The bacterial diversity of the continental shelf sediment in the Yellow Sea was investigated by the cloning and sequencing of PCR-amplified 16S rRNA genes. The majority of the cloned sequences were distinct phylotypes that were novel at the species level. The richness estimator indicated that the sediment sample might harbor up to 32 phylum-level taxa. A large number of low-abundance, phylum-level taxa accounted for most of the observed phylogenetic diversity at our study site, suggesting that these low-abundance taxa might play crucial roles in the shelf sediment ecosystem.

A report of 12 unrecorded bacterial species isolated from Suncheon Bay in Korea

  • Seok Won Jang;Jung Hye Eom;Sanghwa Park
    • Korean Journal of Environmental Biology
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    • v.40 no.4
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    • pp.405-412
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    • 2022
  • Suncheon Bay Ecological Park, possessing abundant fisheries and biological diversity, was registered as a Ramsar wetland in Korea. Approximately 300 bacterial strains were isolated from the Suncheon Bay in a comprehensive study of indigenous prokaryotic species conducted during 2019-2020 in South Korea. A total of 12 bacterial strains were identified using 16S rRNA gene sequencing, demonstrating >98.7% sequence similarity with validly published species. These species were determined to be unrecorded bacterial species in Korea. A total of six strains were isolated from brackish water and Phragmites communis Trin (reed) species. These unrecorded species were phylogenetically diverse and belonged to three classes, six orders, and ten genera. Regarding the genus and class levels, the previously unrecorded species belonged to Jiella, Martelella, Rhizobium, Paracoccus, Rhodovulum, and Altererythrobacter of the class Alphaproteobacteria; Mycolicibacterium, Demequina, and Microbacterium of the class Actinobacteria; Confluentibacter of the class Flavobacteria. The twelve species were further characterized by gram staining, colony and cell morphology, biochemical properties, and phylogenetic position.

Soil Bacterial Community in Red Pine Forest of Mt. Janggunbong, Bonghwa-Gun, Gyeongbuk, Korea, Using Next Generation Sequencing (차세대염기서열방법을 이용한 경북 봉화군 장군봉 소나무림의 토양 박테리아 군집 구성)

  • Lee, Byeong-Ju;Eo, Soo Hyung
    • Journal of Korean Society of Forest Science
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    • v.106 no.2
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    • pp.121-129
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    • 2017
  • The soil microbiome plays important roles in material cycling and plant growth in forest ecosystem. Although a lot of researches on forest soil fungi in Korea have been performed, the studies on forest soil bacterial communities have been limited. In this study, we conducted next generation sequencing (NGS) targeting 16S rRNA gene to investigate the soil bacterial communities from natural red pine (Pinus densiflora) forest in Mt. Janggunbong, Bonghwa-gun, Gyeongbuk, Korea. Our results showed that the entire bacterial communities in the study sites include the phyla Proteobacteria, Acidobacteria, Actinobacteria, Planctomycetes, which have been typically observed in forest soils. The composition ratio of Proteobacteria was the highest in the soil bacteria community. The results reflect that Proteobacteria is copiotroph, which generally favors relatively nutrient-rich conditions with abundant organic matter. Some rhizobia species such as Burkholderia, Bradyrhizobium, Rhizobium, which are known to contribute to soil nitrogen-fixation, exist in the study sites. As a result of correlation analysis between soil physicochemical characteristics and bacteria communities, the soil pH was significantly correlated with the soil bacteria compositions.

Analysis of Microbial Communities Using Culture-dependent and Culture-independent Approaches in an Anaerobic/Aerobic SBR Reactor

  • Lu Shipeng;Park Min-Jeong;Ro Hyeon-Su;Lee Dae-Sung;Park Woo-Jun;Jeon Che-Ok
    • Journal of Microbiology
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    • v.44 no.2
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    • pp.155-161
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    • 2006
  • Comparative analysis of microbial communities in a sequencing batch reactor which performed enhanced biological phosphorus removal (EBPR) was carried out using a cultivation-based technique and 16S rRNA gene clone libraries. A standard PCR protocol and a modified PCR protocol with low PCR cycle was applied to the two clone libraries of the 16S rRNA gene sequences obtained from EBPR sludge, respectively, and the resulting 424 clones were analyzed using restriction fragment length polymorphisms (RFLPs) on 16S rRNA gene inserts. Comparison of two clone libraries showed that the modified PCR protocol decreased the incidence of distinct fragment patterns from about 63 % (137 of 217) in the standard PCR method to about 34 % (70 of 207) under the modified protocol, suggesting that just a low level of PCR cycling (5 cycles after 15 cycles) can significantly reduce the formation of chimeric DNA in the final PCR products. Phylogenetic analysis of 81 groups with distinct RFLP patterns that were obtained using the modified PCR method revealed that the clones were affiliated with at least 11 phyla or classes of the domain Bacteria. However, the analyses of 327 colonies, which were grouped into just 41 distinct types by RFLP analysis, showed that they could be classified into five major bacterial lineages: ${\alpha},\;{\beta},\;{\gamma}-$ Proteobacteria, Actinobacteria, and the phylum Bacteroidetes, which indicated that the microbial community yielded from the cultivation-based method was still much simpler than that yielded from the PCR-based molecular method. In this study, the discrepancy observed between the communities obtained from PCR-based and cultivation-based methods seems to result from low culturabilities of bacteria or PCR bias even though modified culture and PCR methods were used. Therefore, continuous development of PCR protocol and cultivation techniques is needed to reduce this discrepancy.

Microbial Community Diversity in Anaerobic Reactors Digesting Turkey, Chicken, and Swine Wastes

  • Ziganshina, Elvira E.;Belostotskiy, Dmitry E.;Shushlyaev, Roman V.;Miluykov, Vasili A.;Vankov, Petr Y.;Ziganshin, Ayrat M.
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1464-1472
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    • 2014
  • The microbial community structures of two continuous stirred tank reactors digesting turkey manure with pine wood shavings as well as chicken and swine manure were investigated. The reactor fed with chicken/swine wastes displayed the highest organic acids concentration (up to 15.2 g/l) and ammonia concentration (up to 3.7 g/l ammonium nitrogen) and generated a higher biogas yield (up to $366ml/g_{VS}$) compared with the reactor supplied with turkey wastes (1.5-1.8 g/l of organic acids and 1.6-1.7 g/l of ammonium levels; biogas yield was up to $195ml/g_{VS}$). The microbial community diversity was assessed using both sequencing and profiling terminal restriction fragment length polymorphisms of 16S rRNA genes. Additionally, methanogens were analyzed using methyl coenzyme M reductase alpha subunit (mcrA) genes. The bacterial community was dominated by members of unclassified Clostridiales with the prevalence of specific clostridial phylotypes in each reactor, indicating the effect of the substrate type on the community structure. Of the methanogenic archaea, methanogens of the genus Methanosarcina were found in high proportions in both reactors with specific methanosarcinas in each reactor, whereas the strict hydrogenotrophic methanogens of Methanoculleus sp. were found at significant levels only in the reactor fed with chicken/swine manure (based on the analyses of 16S rRNA gene). This suggests that among methanogenic archaea, Methanosarcina species which have different metabolic capabilities, including aceticlastic and hydrogenotrophic methanogenesis, were mainly involved in anaerobic digestion of turkey wastes.

Characteristics of Enteric Pathogens Isolated from Acute Enteritis Patients with Overseas Travel (해외여행 후 발생한 급성장염의 원인 병원체 분석)

  • Choi, Jihye;Lee, Hyunah;Lee, Dayeon;Park, Junhyuk
    • Journal of Environmental Health Sciences
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    • v.48 no.3
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    • pp.176-182
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    • 2022
  • Background: The risk of imported infectious diseases has been increasing with the annual rise in the number of international travelers. Objectives: This study aims to analyze the distribution and characteristics of intestinal bacteria isolated in 2019 from residents of Chungcheongnam-do Province with experience of travelling overseas. Methods: Twenty-three former overseas travelers with diarrhea were analyzed to detect viruses and bacteria according to the Manual for Detection of Foodborne Pathogens at Outbreaks. Additionally, antibiotic susceptibility tests and 16s rRNA sequencing were performed. Results: Twenty-five strains of ten pathogens were isolated from 18 samples. Pathogenic E. coli was the most common at 57.7%, followed by Clostridium perfringens (15.4%), Campylobacter spp. (7.7%), and Salmonella spp. (7.7%). The serotype of Salmonella was confirmed as Salmonella Braenderup, II 9,46:g,[m],[s],t:[e,n,x]. Conclusions: It was confirmed that the major enteric bacterial pathogens isolated from overseas travelers in Chungcheongnam-do Province were pathogenic E. coli, as found in other studies. The study on Plesiomonas shigelloides is meaningful in that it is reported as a rare case of infection in Korea. Antibiotic resistance and 16s rRNA analysis were performed, which is expected to provide important basic data for the prevention of traveler's diarrhea.