• Title/Summary/Keyword: 16S rRNA analysis

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Dark Fermentative Hydrogen Production using the Wastewater Generated from Food Waste Recycling Facilities (혐기 발효 공정을 통한 음식물류 폐기물 탈리액으로부터 수소 생산)

  • Kim, Dong-Hoon;Lee, Mo-Kwon;Lim, So-Young;Kim, Mi-Sun
    • Journal of Hydrogen and New Energy
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    • v.22 no.3
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    • pp.326-332
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    • 2011
  • The authors examined the effects of operating parameters on the $H_2$ production by dark fermentation of the wastewater generated from food waste recycling facilities, in short "food waste wastewater (FWW)". Central composite design based response surface methodology was applied to analyze the effect of initial pH (5.5-8.5) and substrate concentration (2-20 g Carbo. COD/L) on $H_2$ production. The experiment was conducted under mesophilic ($35^{\circ}C$) condition and a heat-treated ($90^{\circ}C$ for 20min)anaerobic digester sludge was used as a seeding source. Although there was a little difference in carbohydrate removal, $H_2$ yield was largely affected by the experimental conditions, from 0.38 to 1.77 mol $H_2$/mol $hexose_{added}$. By applying regression analysis, $H_2$ yield was well fitted based on the coded value to a second order polynomial equation (p = 0.0243): Y = $1.78-0.17X_1+0.30X_2+0.37X_1X_2-0.29X_1{^2}-0.35X_2{^2}$, where $X_1$, $X_2$, and Y are pH, substrate concentration (g Carbo. COD/L), and hydrogen yield (mol $H_2$/mol $hexose_{added}$), respectively. The 2-D response surface clearly showed a high inter-dependency between initial pH and substrate concentration, and the role of these two factors was to control the pH during fermentation. According to the statistical optimization, the optimum condition of initial pH and substrate concentration were 7.0 and 13.4 g Carbo. COD/L, respectively, under which predicted $H_2$ yield was 1.84 mol $H_2$/mol $hexose_{added}$. Microbial analysis using 16S rRNA PCR-DGGE showed that $Clostridium$ sp. such as $Clostridium$ $perfringens$, $Clostridium$ $sticklandii$, and $Clostridium$ $bifermentans$ were main $H_2$-producers.

Isolation and Characteristics of Exopolysaccharide Producing Bacteria in a Ginseng Root System (인삼 근계로부터 다당 생성세균의 분리 및 특성)

  • Cho, Geon-Yeong;Jeon, In-Hwa;Han, Song-Ih;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.297-300
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    • 2013
  • EPS producing bacteria were enumerated in ginseng root system (rhizosphere soil, rhizoplane, inside of root). EPS producing bacterial density of rhizosphere soil, rhizoplane and inside of root were distributed $9.0{\times}10^6$ CFU/g, $7.0{\times}10^6$ CFU/g, and $1.4{\times}10^3$ CFU/g, respectively. Phylogenetic analysis of the 24 EPS producing isolates based on the 16S rRNA gene sequences, EPS producing isolates from rhizosphere soil (RS) belong to genus Arthrobacter (6 strains) and Rhizobium (1 strain). EPS producing bacteria from rhizoplane (RP) were Arthrobacter (6 strains), Rhodococcus (1 strain) and Pseudomonas (1 strain). EPS producing bacteria from inside of root (IR) were categorized into Rhzobium (6 strains), Bacillus (1 strain), Rhodococcus (1 strain), and Pseudomonas (1 strain). Phylogenetic analysis indicated that Arthrobacter may be a member of representative EPS producing bacteria from ginseng rhizosphere soil and rhizoplane, and Rhizobium is typical EPS producing isolates from inside of ginseng root. The yield of EPS was 10.0 and 4.9 g/L by Rhizobium sp. 1NP2 (KACC 17637) and Arthrobacter sp. 5MP1 (KACC 17636). The purified EPS were analyzed by Bio-LC and glucose, galactose, mannose and glucosamine were detected. The major EPS sugar of these strains was glucose (72.7-84.9%).

Serological and Molecular Detection of Toxoplasma gondii and Babesia microti in the Blood of Rescued Wild Animals in Gangwon-do (Province), Korea

  • Hong, Sung-Hee;Kim, Hee-Jong;Jeong, Young-Il;Cho, Shin-Hyeong;Lee, Won-Ja;Kim, Jong-Tak;Lee, Sang-Eun
    • Parasites, Hosts and Diseases
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    • v.55 no.2
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    • pp.207-212
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    • 2017
  • Infections of Toxoplasma gondii and Babesia microti are reported in many wild animals worldwide, but information on their incidence and molecular detection in Korean wild fields is limited. In this study, the prevalence of T. gondii and B. microti infection in blood samples of 5 animal species (37 Chinese water deer, 23 raccoon dogs, 6 roe deer, 1 wild boar, and 3 Eurasian badgers) was examined during 2008-2009 in Gangwon-do (Province), the Republic of Korea (=Korea) by using serological and molecular tests. The overall seropositivity of T. gondii was 8.6% (6/70); 10.8% in Chinese water deer, 4.3% in raccoon dogs, and 16.7% in roe deer. PCR revealed only 1 case of T. gondii infection in Chinese water deer, and phylogenic analysis showed that the positive isolate was practically identical to the highly pathogenetic strain type I. In B. microti PCR, the positive rate was 5.7% (4/70), including 2 Chinese water deer and 2 Eurasian badgers. Phylogenetic analysis results of 18S rRNA and the ${\beta}$-tubulin gene showed that all positive isolates were US-type B. microti. To our knowledge, this is the first report of B. microti detected in Chinese water deer and Eurasian badger from Korea. These results indicate a potentially high prevalence of T. gondii and B. microti in wild animals of Gangwon-do, Korea. Furthermore, Chinese water deer might act as a reservoir for parasite infections of domestic animals.

Differential Impacts on Bacterial Composition and Abundance in Rhizosphere Compartments between Al-Tolerant and Al-Sensitive Soybean Genotypes in Acidic Soil

  • Wen, Zhong-Ling;Yang, Min-Kai;Fazal, Aliya;Liao, Yong-Hui;Cheng, Lin-Run;Hua, Xiao-Mei;Hu, Dong-Qing;Shi, Ji-Sen;Yang, Rong-Wu;Lu, Gui-Hua;Qi, Jin-Liang;Hong, Zhi;Qian, Qiu-Ping;Yang, Yong-Hua
    • Journal of Microbiology and Biotechnology
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    • v.30 no.8
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    • pp.1169-1179
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    • 2020
  • In this study, two soybean genotypes, i.e., aluminum-tolerant Baxi 10 (BX10) and aluminumsensitive Bendi 2 (BD2), were used as plant materials and acidic red soil was used as growth medium. The soil layers from the inside to the outside of the root are: rhizospheric soil after washing (WRH), rhizospheric soil after brushing (BRH) and rhizospheric soil at two sides (SRH), respectively. The rhizosphere bacterial communities were analyzed by high-throughput sequencing of V4 hypervariable regions of 16S rRNA gene amplicons via Illumina MiSeq. The results of alpha diversity analysis showed that the BRH and SRH of BX10 were significantly lower in community richness than that of BD2, while the WRH exhibited no significant difference between BX10 and BD2. Among the three sampling compartments of the same soybean genotype, WRH had the lowest community richness and diversity while showing the highest coverage. Beta diversity analysis results displayed no significant difference for any compartment between the two genotypes, or among the three different sampling compartments for any same soybean genotype. However, the relative abundance of major bacterial taxa, specifically nitrogen-fixing and/or aluminum-tolerant bacteria, was significantly different in the compartments of the BRH and/or SRH at phylum and genus levels, indicating genotype-dependent variations in rhizosphere bacterial communities. Strikingly, as compared with BRH and SRH, the WRH within the same genotype (BX10 or BD2) always had an enrichment effect on rhizosphere bacteria associated with nitrogen fixation.

Antibacterial Activity and Probiotic Potential of Lactobacillus plantarum HKN01: A New Insight into the Morphological Changes of Antibacterial Compound-Treated Escherichia coli by Electron Microscopy

  • Sharafi, Hakimeh;Maleki, Hadi;Ahmadian, Gholamreza;Zahiri, Hossein Shahbani;Sajedinejad, Neda;Houshmand, Behzad;Vali, Hojatollah;Noghabi, Kambiz Akbari
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.225-236
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    • 2013
  • Among several bacteria examined, an antibacterial-producing Lactobacillus strain with probiotic characteristics was selected and identified based on 16S rRNA gene sequencing. Subsequent purification and mode of action of the antibacterial compounds on target cells including E. coli were investigated. Maximum production of the antibacterial compound was recorded at 18 h incubation at $30^{\circ}C$. Interestingly, antibacterial activity remained unchanged after heating at $121^{\circ}C$ for 45 min, 24 h storage in temperature range of $70^{\circ}C$ to room temperature, and 15 min exposure to UV light, and it was stable in the pH of range 2-10. The active compounds were inactivated by proteolytic enzymes, indicating their proteinaceous nature, and, therefore, referred to as bacteriocin-like inhibitory substances. Isolation and partial purification of the effective agent was done by performing ammonium sulfate precipitation and gel filtration chromatography. The molecular mass of the GFC-purified active compound (~3 kDa) was determined by Tris-Tricine SDS-PAGE. To predict the mechanisms of action, transmission electron microscopy (TEM) analysis of ultrathin sections of E. coli before and after antibacterial treatment was carried out. TEM analysis of antibacterial compounds-treated E. coli demonstrated that the completely altered bacteria appear much darker compared with the less altered bacteria, suggesting a change in the cytoplasmic composition. There were also some membrane-bound convoluted structures visible within the completely altered bacteria, which could be attributed to the response of the E. coli to the treatment with the antibacterial compound. According to the in vivo experiments oral administration of L. plantarum HKN01 resulted in recovery of infected BALB/c mice with Salmonella enterica ser. Typhimurium.

Sphingomonas abietis sp. nov., an Endophytic Bacterium Isolated from Korean Fir

  • Lingmin Jiang;Hanna Choe;Yuxin Peng;Doeun Jeon;Donghyun Cho;Yue Jiang;Ju Huck Lee;Cha Young Kim;Jiyoung Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1292-1298
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    • 2023
  • PAMB 00755T, a bacterial strain, was isolated from Korean fir leaves. The strain exhibits yellow colonies and consists of Gram-negative, non-motile, short rods or ovoid-shaped cells. It displays optimal growth conditions at 20℃, 0% NaCl, and pH 6.0. Results of 16S rRNA gene-based phylogenetic analyses showed that strain PAMB 00755T was most closely related to Sphingomonas chungangi MAH-6T (97.7%) and Sphingomonas polyaromaticivorans B2-7T (97.4%), and ≤96.5% sequence similarity to other members of the genus Sphingomonas. The values of average nucleotide identity (79.9-81.3%), average amino acid identity (73.3-75.9%), and digital DNA-DNA hybridization (73.3-75.9%) were significantly lower than the threshold values for species boundaries; these overall genome-related indexes (OGRI) analyses indicated that the strain represents a novel species. Genomic analysis revealed that the strain has a 4.4-Mbp genome encoding 4,083 functional genes, while the DNA G+C content of the whole genome is 66.1%. The genome of strain PAMB 00755T showed a putative carotenoid biosynthetic cluster responsible for its antioxidant activity. The respiratory quinone was identified as ubiquinone 10 (Q-10), while the major fatty acids in the profile were identified as C18:1ω7c and/or C18:1ω6c (summed feature 8). The major polar lipids of strain PAMB 00755T were diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, and phosphatidylcholine. Based on a comprehensive analysis of genomic, phenotypic, and chemotaxonomic characteristics, we proposed the name Sphingomonas abietis sp. nov. for this novel species, with PAMB 00755T as the type strain (= KCTC 92781T = GDMCC 1.3779T).

Amelioration of colitis progression by ginseng-derived exosome-like nanoparticles through suppression of inflammatory cytokines

  • Jisu Kim;Shuya Zhang ;Ying Zhu;Ruirui Wang;Jianxin Wang
    • Journal of Ginseng Research
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    • v.47 no.5
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    • pp.627-637
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    • 2023
  • Background: Damage to the healthy intestinal epithelial layer and regulation of the intestinal immune system, closely interrelated, are considered pivotal parts of the curative treatment for inflammatory bowel disease (IBD). Plant-based diets and phytochemicals can support the immune microenvironment in the intestinal epithelial barrier for a balanced immune system by improving the intestinal microecological balance and may have therapeutic potential in colitis. However, there have been only a few reports on the therapeutic potential of plant-derived exosome-like nanoparticles (PENs) and the underlying mechanism in colitis. This study aimed to assess the therapeutic effect of PENs from Panax ginseng, ginseng-derived exosome-like nanoparticles (GENs), in a mouse model of IBD, with a focus on the intestinal immune microenvironment. Method: To evaluate the anti-inflammatory effect of GENs on acute colitis, we treated GENs in Caco2 and lipopolysaccharide (LPS) -induced RAW 264.7 macrophages and analyzed the gene expression of proinflammatory cytokines and anti-inflammatory cytokines such as TNF-α, IL-6, and IL-10 by real-time PCR (RT-PCR). Furthermore, we further examined bacterial DNA from feces and determined the alteration of gut microbiota composition in DSS-induced colitis mice after administration of GENs through 16S rRNA gene sequencing analysis. Result: GENs with low toxicity showed a long-lasting intestinal retention effect for 48 h, which could lead to effective suppression of pro-inflammatory cytokines such as TNF-α and IL-6 production through inhibition of NF-κB in DSS-induced colitis. As a result, it showed longer colon length and suppressed thickening of the colon wall in the mice treated with GENs. Due to the amelioration of the progression of DSS-induced colitis with GENs treatment, the prolonged survival rate was observed for 17 days compared to 9 days in the PBS-treated group. In the gut microbiota analysis, the ratio of Firmicutes/Bacteroidota was decreased, which means GENs have therapeutic effectiveness against IBD. Ingesting GENs would be expected to slow colitis progression, strengthen the gut microbiota, and maintain gut homeostasis by preventing bacterial dysbiosis. Conclusion: GENs have a therapeutic effect on colitis through modulation of the intestinal microbiota and immune microenvironment. GENs not only ameliorate the inflammation in the damaged intestine by downregulating pro-inflammatory cytokines but also help balance the microbiota on the intestinal barrier and thereby improve the digestive system.

Production of a Fermented Korean Pear Puree using a New Strain Leuconostoc mesenteroides KACC 91495P Isolated from Kimchi (김치에서 신규 Leuconostoc mesenteroides KACC 91495P 균주의 분리 및 이를 이용한 배 발효물의 제조)

  • In, Man-Jin;Kim, Hye-Min;Jin, Hea-Jin;Kim, Dong-Chung;Oh, Nam-Soon;Chae, Hee-Jeong
    • Journal of Applied Biological Chemistry
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    • v.53 no.1
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    • pp.51-55
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    • 2010
  • A lactic acid bacterial strain showing fast growth and high acid production in Korean pear puree was isolated from Kimchi. This strain was analyzed by API 50 CHL kit and 16S rRNA sequencing analysis and identified as Leuconostoc (Ln.) mesenteroides KACC 91495P. Korean pear puree was fermented using Ln. mesenteroides KACC 91495P strain at $30^{\circ}C$ for 18 h. The changes of pH, titratable acidity and viable cell number during fermentation were investigated. The pH and titratable acidity were reached to pH 3.86 and 1.09% after 18 h fermentation, respectively. The viable cell population of Ln. mesenteroides KACC 91495P was rapidly increased to $2.0{\times}10^9\;CFU/g$ during the 9 h of cultivation. The contents of lactic acid, acetic acid and malic acid were determined to be 0.213, 0.259, and 0.217% after 18 h fermentation, respectively. The content of polyphenolic compounds, known as antioxidants, in pear puree were enhanced by Ln. mesenteroides KACC 91495P cultivation. The level of total polyphenolic compounds was increased to around 140% of initial concentration. When the fermented pear puree was kept at $4^{\circ}C$, pH, titratable acidity and number of viable cells population were nearly maintained for 13 days.

Biocontrol Potential of Streptomyces griseus H7602 Against Root Rot Disease (Phytophthora capsici) in Pepper

  • Nguyen, Xuan-Hoa;Naing, Kyaw-Wai;Lee, Young-Seong;Tindwa, Hamisi;Lee, Geon-Hyoung;Jeong, Byoung-Kon;Ro, Hee-Myeong;Kim, Sang-Jun;Jung, Woo-Jin;Kim, Kil-Yong
    • The Plant Pathology Journal
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    • v.28 no.3
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    • pp.282-289
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    • 2012
  • The root rot of pepper (Capsicum annuum L.) caused by Phytophthora capsici is one of the most important diseases affecting this crop worldwide. This work presents the evaluation of the capacity of Streptomyces griseus H7602 to protect pepper plants against Phytophthora capsici and establishes its role as a biocontrol agent. In this study, we isolated an actinomycete strain H7602 from rhizosphere soil, identified it as Streptomyces griseus by 16S rRNA analysis and demonstrated its antifungal activity against various plant pathogens including P. capsici. H7602 produced lytic emzymes such as chitinase, ${\beta}$-1,3-glucanase, lipase and protease. In addition, crude extract from H7602 also exhibited destructive activity toward P. capsici hyphae. In the pot trial, results showed the protective effect of H7602 against pepper from P. capsici. Application of H7602 culture suspension reduced 47.35% of root mortality and enhanced growth of pepper plants for 56.37% in fresh root and 17.56% g in fresh shoot as compared to control, resulting in greater protection to pepper plants against P. capsici infestation. Additionally, the enzymatic activities, chitinase and ${\beta}$-1,3-glucanase, were higher in rhizosphere soil and roots of pepper plants treated with H7602 than other treated plants. Therefore, our results indicated a clear potential of S. griseus H7602 to be used for biocontrol of root rot disease caused by P. capsici in pepper.

Changes of Biological and Chemical Properties during Composting of Livestock Manure with Isolated Native Microbe (토착미생물별 가축분 퇴비화 과정중 생물화학적 특성 변화)

  • Han, Hyo-Shim;Lee, Kyung-Dong
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.6
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    • pp.1126-1135
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    • 2012
  • In order to produce high-quality fermenting composts, bacteria strains with high activities of extracellular enzymes (cellulase, chitinase, amylase, protease and lipase) were isolated from the soils in 6 provinces of Korea, and characterized by 16S rRNA gene sequence analysis and properties. The selected 7 stains inoculated to livestock manure for 2' fermenting time, and experimental treatment divided into 3 groups, B1, B2 and B3, according to microbial activity and enzyme type. Our results showed that microbe applications (B1, B2 and B3) can increase (p<0.05) both rhizomes (17-38%) and enzyme activities (50-81%) in compost after fermenting time, respectively, compared to non-microbe treatment (control). The microbe application also decreased significantly (p<0.05) the $NH_3$ and $H_2S$ gas contents 13.4 and 27.3% compared with control, and the Propionic acid and Butyric acid gas contents 14.5 and 19.6%, respectively, as compared to the control. The microbial degradation rate (%) of pesticides and heavy metals increased significantly (p<0.05) after fermenting time, respectively, as compared to the control. Especially, microbe applications were more effective in total rhizomes yields and bioactivities than non-microbe treatment. Thus the results of this study could help in development of potential bioinoculants and composting techniques that maybe suitable for crop production, and protectable for earth environment under various conditions.