• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.21 no.7
• /
• pp.312-320
• /
• 2020

The objective of the study was to undertake a phylogenetic diversity census of ruminal archaea based on a meta-analysis of 16S rRNA gene sequences that were publicly available in the Ribosomal Database Project. A total of 8,416 sequences were retrieved from the Ribosomal Database Project (release 11, update 5) and included in the construction of a taxonomy tree. Species-level operational taxonomic units (OTUs) were analyzed at a 97% sequence similarity by using the QIIME program. Of the 8,416 sequences, 8,412 were classified into one of three phyla; however, the remaining four sequences could not be classified into a known phylum. The Euryarchaeota phylum was predominant and accounted for 99.8% of the archaeal sequences examined. Among the Euryarchaeota, 65.4% were assigned to Methanobrevibacter, followed by Methanosphaera (10.4%), Methanomassillicoccus (10.4%), Methanomicrobium (7.9%), Methanobacterium (1.9%), Methanimicrococcus (0.5%), Methanosarcina (0.1%), and Methanoculleus (0.1%). The 7,544 sequences that had been trimmed to the V2 and V3 regions clustered into 493 OTUs. Only 17 of those 493 OTUs were dominant groups and accounted for more than 1% of the 7,544 sequences. These results can help guide future research into the dominant ruminal methanogens that significantly contribute to methane emissions from ruminants, research that may lead to the development of anti-methanogenic compounds that inhibit these methanogens regardless of diet or animal species.

• Environmental Engineering Research
• /
• v.14 no.1
• /
• pp.3-9
• /
• 2009

Microorganisms play an important role in the geochemical cycles, industry, environmental cleanup, and biotechnology among other fields. Given the high microbial diversity, identification of the microorganism is essential in understanding and managing the processes. One of the most popular and powerful method for microbial identification is comparative 16S rRNA gene analysis. Due to the highly conserved nature of this essential gene, sequencing and later comparison of it against known rRNA databases can provide assignment of the bacteria into the taxonomy, and the identity of its closest relatives. Isolation and sequencing of 16S rRNA genes directly from natural environments (either from DNA or RNA) can also be used to study the structure of the whole microbial community. Nowadays, novel sequencing technologies with massive outputs are giving researchers worldwide the chance to study the microbial world with a depth that was previously too expensive to achieve. In this article we describe commonly used research approaches for the study of individual microorganisms and microbial communities using the tools provided by Ribosomal Database Project website.

• Journal of Environmental Health Sciences
• /
• v.35 no.5
• /
• pp.393-401
• /
• 2009

We isolated and identified a microorganism which was excellent for ammonia oxidation in the biological control of ammonia gas in odor producing materials from organic composting. The isolated strain was tested for growth characteristics and ammonia elimination efficiency under various conditions of temperature, pH, carbon concentration and ammonia concentration. The strain was isolated from a culture broth used in a $NO_2$ producing test with Griess-Ilosvay reagent. The results of 16S rRNA sequence from the isolated strain by using BLANST (Basic Local Alignment Search Tool) and confirming RDP (Ribosomal Database Project II) and ERRD (The European Ribosomal RNA Database) indicate that the strain is related to Delftia sp. UV-Spectrophotometer (Shimadzu, UVmini-1240) was used as a microbial growth test by measuring turbidity on OD660nm and ammonia concentration was measured by Spectrophotometer (HACH, DR-4000). The optimum growth culture conditions of the ammonia oxidizer Delftia sp. were $30^{\circ}C$, pH 7, glucose concentration 1.00% and $(NH_4)_2SO_4$ 0.5 g/l. Ammonia elimination efficiency was over 94% under the same conditions.

• Journal of Animal Reproduction and Biotechnology
• /
• v.34 no.3
• /
• pp.157-165
• /
• 2019

This study was conducted to analyze the diversity census of fecal microbiome in horses using meta-analysis of equine 16S rRNA gene sequences that are available in the Ribosomal Database Project (RDP; Release 11, Update 5). The search terms used were "horse feces (or faeces)" and "equine feces (or faeces)". A total of 842 sequences of equine feces origin were retrieved from the RDP database, where 744 sequences were assigned to 10 phyla placed within Domain Bacteria. Firmicutes (n = 391) and Bacteroidetes (n = 203) were the first and the second dominant phyla, respectively, followed by Verrucomicrobia (n = 58), Proteobacteria (n = 30) and Fibrobacteres (n = 24). Clostridia (n = 319) was the first dominant class placed within Bacteroidetes while Bacteroidia (n = 174) was the second dominant class placed within Bacteroidetes. The remaining 98 sequences were assigned to phylum Euryarchaeota placed within Domain Archaea, where 74 sequences were assigned to class Methanomicrobia. The current results will improve understanding of the diversity of fecal microbiome in horses and may be used to further analyze equine fecal microbiome in future studies.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.18 no.12
• /
• pp.466-472
• /
• 2017

This study was designed to examine the diversity census of fungi in rumen microbiome via meta-analysis of fungal 28S rDNA sequences. Both terms, "rumen" and "ruminal," were searched to retrieve the sequences of rumen fungi. As of September 2016, these sequences (n=165) of ruminal origin were retrieved from the Ribosomal Database Project (RDP; http://rdp.cme.msu.edu), an archive of all 28S rDNA sequences and were assigned to the phyla Ascomycota, Neocallimastigomycota, and Basidiomycota, which accounted for 109, 48, and 8 of the 165 sequences, respectively. Ascomycota sequences were assigned to the genera Pseudonectria, Magnaporthe, Alternaria, Cochliobolus, Cladosporium, and Davidiella, including fungal plant pathogens or mycotoxigenic species. Moreover, Basidiomycota sequences were assigned to the genera Thanatephorus and Cryptococcus, including fungal plant pathogens. Furthermore, Neocallimastigomycota sequences were assigned to the genera Cyllamyces, Neocallimastix, Anaeromyces, Caecomyces, Orpinomyces, and Piromyces, which may degrade the major structural carbohydrates of the ingested plant material. This study provided a collective view of the rumen fungal diversity using a meta-analysis of 28S rDNA sequences. The present results will provide a direction for further studies on ruminal fungi and be applicable to the development of new analytic tools.

• Restorative Dentistry and Endodontics
• /
• v.36 no.6
• /
• pp.498-505
• /
• 2011

Objectives: The purpose of this in vivo study was to investigate the microbial diversity in symptomatic and asymptomatic canals with primary endodontic infections by using GS FLX Titanium pyrosequencing. Materials and Methods: Sequencing was performed on 6 teeth (symptomatic, n = 3; asymptomatic, n = 3) with primary endodontic infections. Amplicons from hypervariable region of the small-subunit ribosomal RNA gene were generated by polymerized chain reaction (PCR), and sequenced by means of the GS FLX Titanium pyrosequencing. Results: On average, 10,639 and 45,455 16S rRNA sequences for asymptomatic and symptomatic teeth were obtained, respectively. Based on Ribosomal Database Project Classifier analysis, pyrosequencing identified the 141 bacterial genera in 13 phyla. The vast majority of sequences belonged to one of the seven phyla: Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, Proteobacteria, Spirochetes, and Synergistetes. In genus level, Pyramidobacter, Streptococcus, and Leptotrichia constituted about 50% of microbial profile in asymptomatic teeth, whereas Neisseria, Propionibacterium, and Tessaracoccus were frequently found in symptomatic teeth (69%). Grouping the sequences in operational taxonomic units (3%) yielded 450 and 1,997 species level phylotypes in asymptomatic and symptomatic teeth, respectively. The total bacteria counts were significantly higher in symptomatic teeth than that of asymptomatic teeth (p < 0.05). Conclusions: GS FLX Titanium pyrosequencing could reveal a previously unidentified high bacterial diversity in primary endodontic infections.

• Microbiology and Biotechnology Letters
• /
• v.32 no.3
• /
• pp.218-223
• /
• 2004

Myxobacteria are Gram-negative soil bacteria known to be a rich source of potentially useful secondary metabolites. We have isolated 204 strains of bacteriolytic myxobacteria from soil samples collected in Korea and determined their 16S rRNA sequences. Sequence analysis of the partially determined 16S rRNA sequences has suggested that 132 isolates (65% of total isolates) belong to the genus Myxococcus and 59 isolates (29% of total isolates) belong to the genus Corallococcus. Meanwhile, 4 isolates appear to be Archangium spp. and the other 4 isolates appear to be Stigmatella spp. Genera of the remained 5 isolates have not been identified because their 16S rRNA sequences are distantly related to those of known myxobacteria.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.2
• /
• pp.248-255
• /
• 2012

In order to develop a protocol to quantify cyanobacteria and Microcystis simultaneously, the primers and probe were designed from the conserved regions of 16S rRNA gene sequences of cyanobacteria and Microcystis, respectively. Probe match analysis of the Ribosomal Database Project showed that the primers matched with over 97% of cyanobacterial 16S rRNA genes, indicating these can be used to amplify cyanobacteria specifically. The TaqMan probe, which is located between two primers, matched with 98.2% of sequences in genus GpXI, in which most Microcystis strains are included. The numbers of cyanobacterial genes were estimated with the emission of SYBR Green from the amplicons with two primers, whereas those of Microcystis spp. were measured from the fluorescence of CAL Fluor Gold 540 emitted by exonuclease activity of Taq DNA polymerase in amplification. It is expected that this method enhances the accuracy and reduces the time to count cyanobacteria and potential toxigenic Microcystis spp. in aquatic environmental samples.

• The Journal of Pediatrics of Korean Medicine
• /
• v.34 no.4
• /
• pp.43-58
• /
• 2020

Objectives The purpose of this study is to analyze the effect of various herbal medicin on gut microbiota. Methods Electronic searches were performed using NDSL, OASIS, KISS, KMBASE, K-portal, Pub med, Cochrane, CNKI. Results we analyzed 25 experimental studies on the effect of herbal medicine on microbiota. Diabetes, obesity, inflammatory bowel disease have been frequently studied in micobiota-related disease. The most common experimental animal model used in the studies C57BL/7 mouse. Among the studies wherein single herbal medication were used, Gynostemma pentaphyllum was most commonly studies, and different herbal medications were used in the studies wherein complex herbal medications were studied. Next generation sequencing was performed using Illumina MiSeq system, and gut microbiota analysis was performed using QIIME and Ribosomal Database Project (RDP). In most studies, the herbal medicines exerted regulatory effects on gut microbiota and improved the symptoms of the experimental groups. Conclusions This review provides basic data on the correlation between korean medicine and gut microbiota, as well as information for the development of korean medicine.

• Korean Journal of Microbiology
• /
• v.37 no.2
• /
• pp.137-144
• /
• 2001

In order to investigate the diversity of bacterial community in the mud flat of Sunchon Bay, Chunnam province, diversity of amplified 16S rDNA was examined. Total DNA was extracted from sediment soils and 16S rDNAs were amplified using PCR primers based on the universally conserved sequences in bacteria. Clonal libraries were constructed and 111 clones were examined by amplified rDNA restriction analysis (ARDRA) using HaeIII. Clones were clustered based on restriction patterns using computer program, GelCompar II. One hundred different RFLP types were detected from 111 clones. The 20 clones were selected and sequenced according to dendrograms derived from ARDRA, to cover most of the bacterial diversity in the clone libraries. None of the clones were identical to any representatives in the Ribosomal Database Project small subunit RNA databases and GenBank. All sequences showed between 77 and 96.8% similarity to the known 16s rRNA sequence from cultured organisms. The 20 clones sequenced fell into seven major lineages of the domain Bacteria: alpha-, delta-, gamma-Proteobacteria, low G+C Gram positive bacteria, high G+C Gram positive bacteria, Sphingobacteria (Cytophaga) and Cyanobacteria (chloroplast). Among the clones, the Proteobacteria were dominant.