• Title/Summary/Keyword: Microbial profiling

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Fecal Microbiota Profiling of Holstein and Jersey, in South Korea : A Comparative Study (국내에서 사육되는 Holstein 젖소과 Jersey 젖소의 대변 미생물 분석 : 비교연구)

  • Gwangsu Ha;Ji-Won Seo;Hee Gun Yang;Se Won Park;Soo-Young Lee;Young Kyoung Park;RanHee Lee;Do-Youn Jeong;Hee-Jong Yang
    • Journal of Life Science
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    • v.33 no.7
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    • pp.565-573
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    • 2023
  • In light of the complex interactions between the host animal and its resident gut microbiomes, studies of these microbial communities as a means to improve cattle production are important. This study was conducted to analyze the intestinal microorganisms of Holstein (HT) and Jersey (JS), raised in Korea and to clarify the differences in microbial structures according to cattle species through next-generation sequencing. The alpha-diversity analysis revealed that most species richness and diversity indices were significantly higher in JS than in HT whereas phylogenetic diversity, which is the sum of taxonomic distances, is not significant. Microbial composition analysis showed that the intestinal microbial community structure of the two groups differed. In the both groups, a significant correlation was observed among the distribution of several microbes at the family level. In particular, a highly significant correlation (p<0.0001) among a variety of microbial distributions was found in JS. Beta-diversity analyis was to performed to statistically verify whether a difference exists in the intestinal microbial community structure of the two groups. Principal coordinate analysis and unweighted pair group method with arithmetic mean (UPGMA) clustering analysis showed separation between the HT and JS clusters. Meanwhile, permutational multivariate analysis of variance (PERMANOVA) revealed that their microbial structures are significantly different (p<0.0001). LEfSe biomarker analysis was performed to discover the differenc microbial features between the two groups. We found that several microbes, such as Firmicutes, Bacilli, Moraxellaceae and Pseudomonadales account for most of the difference in intestinal microbial community structure between the two groups.

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.

Characterization of Water Quality and Microbial Communities in Rivers in Changwon city (창원시 하천의 수질 및 미생물상 분석)

  • Kim Sun-A;Kim Chung-Hye;Lim Byung-Ran;Cho Kwang-Hyun;Park Hee-Chang;Joo Woo Hong
    • Journal of Life Science
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    • v.16 no.1
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    • pp.148-155
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    • 2006
  • The diversity of bacterial populations in rivers flowing through Changwon City, was investigated using quinone profiling. The physicochemical properties such as temperature, pH, dissolved oxygen(DO), dissolved organic carbon (DOC) and biochemical oxygen demand (BOD) were also measured in this study. Ubiquinone (UQ)-8, UQ-9 and UQ-10 were observed in all samples for the sites investigated. UQ-8 was the -predominant quinone species in rivers except for Namch'on downstream, T'owolch'on, and Kaumchongch'on in autumn, while UQ-8 was also found as major quinones in the sample except for Hanamch'on, T'owolch'on, Kaumchongch'on, and Namsanch'on in winter. A higher concentration of DOC in rivers yield high concentration of plastoquinone (PQ-9) in autumn and those of total quinones in winter, respectively. Correlation analysis also indicate that BOD is considered to be a major factor controlling the concentration of PQ in rivers.

Comparison of the oral microbial composition between healthy individuals and periodontitis patients in different oral sampling sites using 16S metagenome profiling

  • Kim, Yeon-Tae;Jeong, Jinuk;Mun, Seyoung;Yun, Kyeongeui;Han, Kyudong;Jeong, Seong-Nyum
    • Journal of Periodontal and Implant Science
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    • v.52 no.5
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    • pp.394-410
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    • 2022
  • Purpose: The purpose of this study was to compare the microbial composition of 3 types of oral samples through 16S metagenomic sequencing to determine how to resolve some sampling issues that occur during the collection of sub-gingival plaque samples. Methods: In total, 20 subjects were recruited. In both the healthy and periodontitis groups, samples of saliva and supra-gingival plaque were collected. Additionally, in the periodontitis group, sub-gingival plaque samples were collected from the deepest periodontal pocket. After DNA extraction from each sample, polymerase chain reaction amplification was performed on the V3-V4 hypervariable region on the 16S rRNA gene, followed by metagenomic sequencing and a bioinformatics analysis. Results: When comparing the healthy and periodontitis groups in terms of alpha-diversity, the saliva samples demonstrated much more substantial differences in bacterial diversity than the supra-gingival plaque samples. Moreover, in a comparison between the samples in the case group, the diversity score of the saliva samples was higher than that of the supra-gingival plaque samples, and it was similar to that of the sub-gingival plaque samples. In the beta-diversity analysis, the sub-gingival plaque samples exhibited a clustering pattern similar to that of the periodontitis group. Bacterial relative abundance analysis at the species level indicated lower relative frequencies of bacteria in the healthy group than in the periodontitis group. A statistically significant difference in frequency was observed in the saliva samples for specific pathogenic species (Porphyromonas gingivalis, Treponema denticola, and Prevotella intermedia). The saliva samples exhibited a similar relative richness of bacterial communities to that of sub-gingival plaque samples. Conclusions: In this 16S oral microbiome study, we confirmed that saliva samples had a microbial composition that was more similar to that of sub-gingival plaque samples than to that of supra-gingival plaque samples within the periodontitis group.

Microbial profiling of peri-implantitis compared to the periodontal microbiota in health and disease using 16S rRNA sequencing

  • Hyun-Joo Kim;Dae-Hee Ahn;Yeuni Yu;Hyejung Han;Si Yeong Kim;Ji-Young Joo;Jin Chung;Hee Sam Na;Ju-Youn Lee
    • Journal of Periodontal and Implant Science
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    • v.53 no.1
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    • pp.69-84
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    • 2023
  • Purpose: The objective of this study was to analyze the microbial profile of individuals with peri-implantitis (PI) compared to those of periodontally healthy (PH) subjects and periodontitis (PT) subjects using Illumina sequencing. Methods: Buccal, supragingival, and subgingival plaque samples were collected from 109 subjects (PH: 30, PT: 49, and PI: 30). The V3-V4 region of 16S rRNA was sequenced and analyzed to profile the plaque microbiota. Results: Microbial community diversity in the PI group was higher than in the other groups, and the 3 groups showed significantly separated clusters in the buccal samples. The PI group showed different patterns of relative abundance from those in the PH and PT groups depending on the sampling site at both genus and phylum levels. In all samples, some bacterial species presented considerably higher relative abundances in the PI group than in the PH and PT groups, including Anaerotignum lactatifermentans, Bacteroides vulgatus, Faecalibacterium prausnitzii, Olsenella uli, Parasutterella excrementihominis, Prevotella buccae, Pseudoramibacter alactolyticus, Treponema parvum, and Slackia exigua. Network analysis identified that several well-known periodontal pathogens and newly recognized bacteria were closely correlated with each other. Conclusions: The composition of the microbiota was considerably different in PI subjects compared to PH and PT subjects, and these results could shed light on the mechanisms involved in the development of PI.

Effective microbial molecular diagnosis of periodontitis-related pathogen Porphyromonas gingivalis from salivary samples using rgpA gene

  • Jinuk Jeong;Yunseok Oh;Junhyeon Jeon;Dong-Heon Baek;Dong Hee Kim;Kornsorn Srikulnath;Kyudong Han
    • Genomics & Informatics
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    • v.21 no.1
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    • pp.13.1-13.8
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    • 2023
  • Importance of accurate molecular diagnosis and quantification of particular disease-related pathogenic microorganisms is highlighted as an introductory step to prevent and care for diseases. In this study, we designed a primer/probe set for quantitative real-time polymerase chain reaction (qRT-PCR) targeting rgpA gene, known as the specific virulence factor of periodontitis-related pathogenic bacteria 'Porphyromonas gingivalis', and evaluated its diagnostic efficiency by detecting and quantifying relative bacterial load of P. gingivalis within saliva samples collected from clinical subjects. As a result of qRT-PCR, we confirmed that relative bacterial load of P. gingivalis was detected and quantified within all samples of positive control and periodontitis groups. On the contrary, negative results were confirmed in both negative control and healthy groups. Additionally, as a result of comparison with next-generation sequencing (NGS)-based 16S metagenome profiling data, we confirmed relative bacterial load of P. gingivalis, which was not identified on bacterial classification table created through 16S microbiome analysis, in qRT-PCR results. It showed that an approach to quantifying specific microorganisms by applying qRT-PCR method could solve microbial misclassification issues at species level of an NGS-based 16S microbiome study. In this respect, we suggest that P. gingivalis-specific primer/probe set introduced in present study has efficient applicability in various oral healthcare industries, including periodontitis-related microbial molecular diagnosis field.

Search for Plant-originated Antibacterial Compounds Against Pathogen (Acidovorax avenae subsp. citrulli) of Watermelon Bacterial Fruit Blotch (수박 과실썩음병 병원균(Acidovorax avenae subsp. citrulli)에 대한 식물유래 항균 활성물질 탐색)

  • Noh, Jin-Taek;Choi, Yong-Hwa
    • Korean Journal of Organic Agriculture
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    • v.23 no.1
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    • pp.77-89
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    • 2015
  • 133 Species of medicinal plants were used for the development of natural agrichemicals with anti-microbial activity against Acidovorax avenae subsp. citrulli, a pathogen of bacterial fruit blotch in watermelon. The MeOH-extracts of these medicinal plants were examined for anti-microbial activity by bioassay. The MeOH-extract of Citrus unshiu Markovich had the strongest antibacterial activity against Acidovorax avenae subsp. citrulli. To identify anti-microbial compounds from Citrus unshiu Markovich, solvent-fractionation was used. The fraction of hexane, which showing the highest value of anti-microbial activity, was analyzed by GC-MS. Each mass spectra, corresponding to each peak of chromatogram, was compared to mass database of Wiley library. As a result, d-Limonene, ${\gamma}$-terpinene, ${\beta}$-linalool, terpineol, palmitic acid, 9,12-octadecadienoic acid, Linolenic acid, and stigmasterol were identified. Among them, d-Limonene, ${\gamma}$-terpinene, ${\beta}$-linalool, and terpineol confirmed to be shown the anti-microbial activity by bioassay. Especially, d-Limonene and ${\gamma}$-terpinene found to have strong activity. In conclusion, we thought d-limonene and ${\gamma}$-terpinene from Citrus unshiu Markovich. Latin, had anti-microbial activity against Acidovorax avenae subsp. citrulli and could be candidates for the control agents for the control of bacterial fruit blotch in watermelon.

The Effects of Genetically Modified Crops on Soil Microbial Community (유전자변형 작물이 토양 미생물상에 미치는 영향)

  • Lee, Ki-Jong;Oh, Sung-Dug;Sohn, Soo-In;Ryu, Tae-Hun;Park, Jong-Sug;Lee, Jang-Yong;Cho, Hyun-Suk;Ahn, Byung-Ohg
    • Korean Journal of Environmental Agriculture
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    • v.31 no.2
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    • pp.192-199
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    • 2012
  • BACKGROUND: Genetically modified (GM) crops must receive relevant regulator's authorization before they can be sold as seed or used food, feed and processing. Before approving any GM crop, the relevant government ministries are required to examine environmental risk assessment to make scientifically sound and socially acceptable decisions. But one of the least studied and understood areas in the environmental risk assessment of GM crops are their impact on soil microbial community. METHODS AND RESULTS: Recently, advanced methods have been developed to characterize the soil microbial community in various environments. In this study, the culture-dependent and culture-independent technical approaches for profiling soil microbial communities are summarized and their applicability to assess GM crops are discussed. CONCLUSION(S): We concluded that the effect of GM crops on soil microbial community need to be assessed on a case by case basis. The combination of culture-dependent and culture-independent method was necessary for reliable and detailed assessment of effect of GM crops on soil microbial community.

Application of Methodology for Microbial Community Analysis to Gas-Phase Biofilters (폐가스 처리용 바이오필터에 미생물 군집 분석 기법의 적용)

  • Lee, Eun-Hee;Park, Hyunjung;Jo, Yun-Seong;Ryu, Hee Wook;Cho, Kyung-Suk
    • Korean Chemical Engineering Research
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    • v.48 no.2
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    • pp.147-156
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    • 2010
  • There are four key factors for gas-phase biofilters; biocatalysts(microorganisms), packing materials, design/operating techniques, and diagnosis/management techniques. Biofilter performance is significantly affected by microbial community structures as well as loading conditions. The microbial studies on biofilters are mostly performed on basis of culture-dependent methods. Recently, advanced methods have been proposed to characterize the microbial community structure in environmental samples. In this study, the physiological, biochemical and molecular methods for profiling microbial communities are reviewed, and their applicability to biofilters is discussed. Community-level physiological profile is based on the utilization capability of carbon substrate by heterotrophic community in environmental samples. Phospholipid fatty acid analysis method is based on the variability of fatty acids present in cell membranes of different microorganisms. Molecular methods using DNA directly extracted from environmental samples can be divided into "partial community DNA analysis" and "whole community DNA analysis" approaches. The former approaches consist in the analysis of PCR-amplified sequence, the genes of ribosomal operon are the most commonly used sequences. These methods include PCR fragment cloning and genetic fingerprinting such as denaturing gradient gel electrophoresis, terminal-restriction fragment length polymorphism, ribosomal intergenic spacer analysis, and random amplified polymorphic DNA. The whole community DNA analysis methods are total genomic cross-DNA hybridization, thermal denaturation and reassociation of whole extracted DNA and extracted whole DNA fractionation using density gradient.

Diversity and Succession of the Bacterial Community during the Initial Fermentation Period in Modernized Soy Sauce (Ganjang) (개량식 간장의 발효 초기 단계에서의 미생물 다양성 및 천이에 관한 연구)

  • Ho Jin Jeong;Gwangsu Ha;Jungmi Lee;Yeji Song;Do-Youn Jeong;Hee-Jong Yang
    • Journal of Life Science
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    • v.33 no.6
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    • pp.481-489
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    • 2023
  • The taste and quality of soy sauce, a fermented liquid condiment, is greatly influenced by microbial metabolism during fermentation. To investigate the microbiological characteristics of ganjang during the initial fermentation process, we prepared meju (fermented soybean) blocks fermented with starter cultures and solar salts and analyzed the microbial community quantitively using 16S rRNA gene profiling from ganjang that had been fermented over a five-week period. The ganjang samples were collected and analyzed after soaking for week one (1W), three (3W), and five (5W) weeks. We found that Halomonadaceae was significantly higher in the 1W group (89.83%) than the 3W and 5W groups (14.46%, and 13.78%, respectively). At a species level, Chromohalobacter beijerinckii and Chromohalobacter canadensis were the dominant species in the 1W group but several taxa such as Bacillus subtilis, Pediococcus acidilactici, and Enterococcus faecalis were more abundant in the 3W and 5W groups. Pearson correlation analysis of the relative abundance of the bacteria showed a negative correlation between Chromohalobacter and two bacterial genera Bacillus and Enterococcus. Beta-diversity showed a statistical distinction between the 1W and the 3W and 5W groups, while no significance was evident between the 3W and 5W groups. Linear discriminant effect size analysis was used to identify biomarkers and significant differences in the relative abundance of several halophilic bacteria, Bacillus sp. and lactic acid bacteria at 1W, 3W, and 5W, recpectively, which indicates the important role of the bacterial community at these time points.