• Asian-Australasian Journal of Animal Sciences
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• 제32권2호
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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.

• ALGAE
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• 제37권1호
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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.

• 대한의생명과학회지
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• 제28권4호
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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.

• Journal of Species Research
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• 제12권2호
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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.

• Journal of Microbiology and Biotechnology
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• 제33권9호
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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.

• Journal of Microbiology and Biotechnology
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• 제17권1호
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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.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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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.

• Journal of Microbiology and Biotechnology
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• 제8권1호
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• pp.67-73
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• 1998

A bacterial strain that excretes hemolysins and proteases into the growth medium was isolated from sea water and designated as KYJ 961. A nearly complete nucleotide sequence of a 16S ribosomal RNA gene from the isolate was determined following the isolation and cloning of amplified genes. On the basis of the 16S ribosomal DNA sequence data, and morphological, chemotaxonomic, and physiological characteristics, strain KYJ 961 was classified as a strain of Bacillus cereus.

• 미생물학회지
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• 제47권2호
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• pp.130-136
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• 2011

러시아의 바이칼에 존재하는 고유종인 Baikalospongia과 Lubomirskia의 sponge로부터 방선균을 분리하였다. 분리된 방선균은 16S rRNA gene 분석 결과, Streptomyces griseoplanus, S. halstedii, S. violascens, S. flavovirens, S. microflavus에 각각 속하였다. 이 방선균들은 온도, 탄소이용, enzyme 활성, fatty acid 조성 등의 실험 결과에서, 각각 서로 다른 특징을 나타냈었다. 분리된 방선균의 배양온도는 30-37, pH는 8-9, 염분농도는 0-1.5에서 가장 잘 자라는 것으로 확인되었다. 주요 cellular fatty acid는 anteiso-$C_{15:0}$, iso-$C_{15:0}$ and iso-$C_{16:0}$로 나타났다. 특히 ATS-BA-19는 DNase와 chitinase 활성을 나타내었고, ATS-BA-16는 cellulase와 protease 활성을 나타내는 것으로 확인되었다. 또 두개의 분리된 방선균에서 그람음성 균주인 Pseudomonas aeruginosa에서 생장을 저해함을 확인하였다.

• 한국동물생명공학회지
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• 제36권1호
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• pp.35-44
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• 2021

A pregnancy diagnosis is an important standard for control of livestock's reproduction in paricular dairy cattle. High reproductive performance in dairy animals is a essential condition to realize of high life-time production. Pregnancy diagnosis is crucial to shortening the calving interval by enabling the farmer to identify open animals so as to treat or re-breed them at the earliest opportunity. MicroRNAs are short RNA molecules which are critically involved in regulating gene expression during both health and disease. This study is sought to establish the feasible of circulating miRNAs as biomarkers of early pregnancy in cattle. We applied Illumina small-RNA sequencing to profile miRNAs in plasma samples collected from 12 non-pregnant cows ("open" cows: samples were collected before insemination (non-pregnant state) and after pregnancy check at the indicated time points) on weeks 0, 4, 8, 12 and 16. Using small RNA sequencing we identified a total of 115 miRNAs that were differentially expressed weeks 16 relative to non-pregnancy ("open" cows). Weeks 8, 12 and 16 of pregnancy commonly showed a distinct increase in circulating levels of miR-221 and miR-320a. Through genome-wide analyses we have successfully profiled plasma miRNA populations associated with pregnancy in cattle. Their application in the field of reproductive biology has opened up opportunities for research communities to look for pregnancy biomarker molecules in dairy cattle.