• Title/Summary/Keyword: 16S rRNA Gene

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Effect of feeding raw potato starch on the composition dynamics of the piglet intestinal microbiome

  • Yi, Seung-Won;Lee, Han Gyu;So, Kyoung-Min;Kim, Eunju;Jung, Young-Hun;Kim, Minji;Jeong, Jin Young;Kim, Ki Hyun;Oem, Jae-Ku;Hur, Tai-Young;Oh, Sang-Ik
    • Animal Bioscience
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    • v.35 no.11
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    • pp.1698-1710
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    • 2022
  • Objective: Raw potato starch (RPS) is resistant to digestion, escapes absorption, and is metabolized by intestinal microflora in the large intestine and acts as their energy source. In this study, we compared the effect of different concentrations of RPS on the intestinal bacterial community of weaned piglets. Methods: Male weaned piglets (25-days-old, 7.03±0.49 kg) were either fed a corn/soybean-based control diet (CON, n = 6) or two treatment diets supplemented with 5% RPS (RPS5, n = 4) or 10% RPS (RPS10, n = 4) for 20 days and their fecal samples were collected. The day 0 and 20 samples were analyzed using a 16S rRNA gene sequencing technology, followed by total genomic DNA extraction, library construction, and high-throughput sequencing. After statistical analysis, five phyla and 45 genera accounting for over 0.5% of the reads in any of the three groups were further analyzed. Furthermore, short-chain fatty acids (SCFAs) in the day 20 fecal samples were analyzed using gas chromatography. Results: Significant changes were not observed in the bacterial composition at the phylum level even after 20 d post feeding (dpf); however, the abundance of Intestinimonas and Barnesiella decreased in both RPS treatment groups compared to the CON group. Consumption of 5% RPS increased the abundance of Roseburia (p<0.05) and decreased the abundance of Clostridium (p<0.01) and Mediterraneibacter (p< 0.05). In contrast, consumption of 10% RPS increased the abundance of Olsenella (p<0.05) and decreased the abundance of Campylobacter (p<0.05), Kineothrix (p<0.05), Paraprevotella (p<0.05), and Vallitalea (p<0.05). Additionally, acetate (p<0.01), butyrate (p<0.05), valerate (p = 0.01), and total SCFAs (p = 0.01) were upregulated in the RPS5 treatment group Conclusion: Feeding 5% RPS altered bacterial community composition and promoted gut health in weaned piglets. Thus, resistant starch as a feed additive may prevent diarrhea in piglets during weaning.

The Gut Microbiota of Pregnant Rats Alleviates Fetal Growth Restriction by Inhibiting the TLR9/MyD88 Pathway

  • Hui Tang;Hanmei Li;Dan Li;Jing Peng;Xian Zhang;Weitao Yang
    • Journal of Microbiology and Biotechnology
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    • v.33 no.9
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    • pp.1213-1227
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    • 2023
  • Fetal growth restriction (FGR) is a prevalent obstetric condition. This study aimed to investigate the role of Toll-like receptor 9 (TLR9) in regulating the inflammatory response and gut microbiota structure in FGR. An FGR animal model was established in rats, and ODN1668 and hydroxychloroquine (HCQ) were administered. Changes in gut microbiota structure were assessed using 16S rRNA sequencing, and fecal microbiota transplantation (FMT) was conducted. HTR-8/Svneo cells were treated with ODN1668 and HCQ to evaluate cell growth. Histopathological analysis was performed, and relative factor levels were measured. The results showed that FGR rats exhibited elevated levels of TLR9 and myeloid differentiating primary response gene 88 (MyD88). In vitro experiments demonstrated that TLR9 inhibited trophoblast cell proliferation and invasion. TLR9 upregulated lipopolysaccharide (LPS), LPS-binding protein (LBP), interleukin (IL)-1β and tumor necrosis factor (TNF)-α while downregulating IL-10. TLR9 activated the TARF3-TBK1-IRF3 signaling pathway. In vivo experiments showed HCQ reduced inflammation in FGR rats, and the relative cytokine expression followed a similar trend to that observed in vitro. TLR9 stimulated neutrophil activation. HCQ in FGR rats resulted in changes in the abundance of Eubacterium_coprostanoligenes_group at the family level and the abundance of Eubacterium_coprostanoligenes_group and Bacteroides at the genus level. TLR9 and associated inflammatory factors were correlated with Bacteroides, Prevotella, Streptococcus, and Prevotellaceae_Ga6A1_group. FMT from FGR rats interfered with the therapeutic effects of HCQ. In conclusion, our findings suggest that TLR9 regulates the inflammatory response and gut microbiota structure in FGR, providing new insights into the pathogenesis of FGR and suggesting potential therapeutic interventions.

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).

Positive effects of grazing on blood components and intestinal microbiota in growing horses

  • Ji Hyun Yoo;Jong An Lee;Jae Young Choi;Sang Min Shin;Moon Cheol Shin;Hyeon Ah Kim;Yong Jun Kang;Hee Chung Ji;In Cheol Cho;Byoung Chul Yang
    • Korean Journal of Agricultural Science
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    • v.49 no.4
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    • pp.963-971
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    • 2022
  • Production of high-quality horses is important to make the horse industry grow. Grazing during the growing period can be an important factor affecting the production of high-quality horses. The objective of this study was to determine the effects of grazing on growing horses by analyzing their blood components and intestinal microbiota. Twelve growing horses for evaluating blood components and ten growing horses for evaluating intestinal microbiota were raised for about seven months and separated by two treatments: grazing vs. stable. Complete blood count, blood chemistry, and creatine kinase levels were analyzed as blood components and a 16s rRNA gene sequence analysis was performed to analyze intestinal microbiota. Calcium ions tended to be lower in the group with grazing treatment. Alkaline phosphatase and creatine kinase tended to be higher in the group with grazing treatment. These results indicate that grazing can provide horses with more exercise than staying in stables. At the phylum level, Firmicutes/Bacteroidetes ratios in grazing and stable groups were 4.2 and 6.5, respectively. Because various studies have reported that a. high Firmicutes/Bacteroidetes ratio indicates obesity, the method of raising horses might affect their physical ability. At the species level, rates of Clostridium butyricum in grazing and stable groups were 3.2% and 13.1%, respectively. Some strains of C. butyricum can cause several diseases such as botulism. These results indicate that grazing can positively affect growing horses in terms of blood components and intestinal microbiota. Moreover, grazing can be helpful to make growing horses healthy through proper exercise.

Soil Chemical Property and Microbial Community under Organic and Conventional Radish Farming Systems (무 유기재배와 관행재배 토양의 화학성과 미생물 군집 비교)

  • Kang, Ho-Jun;Yang, Sung-Nyun;Song, Kwan-Cheol;Cho, Young-Yuen;Kim, Yu-Kyoung
    • Korean Journal of Organic Agriculture
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    • v.27 no.4
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    • pp.479-499
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    • 2019
  • This study was conducted to investigate the responses of soil properties and microbial communities to different agricultural management and soil types, including organic management in Andisols (Org-A), organic management in Non-andisols (Org-NA), conventional management in Andisols (Con-A) and conventional management in Non-andisols (Con-NA) by using a pyrosequencing approach of 16S rRNA gene amplicon in Radish farms of volcanic ash soil in Jeju island. The results showed that agricultural management systems had a little influence on the soil chemical properties but had significant influence on microbial communities. In addition, soil types had significant influences on both the soil chemical properties and microbial communities. Organic farming increased the microbial density of bacteria and biomass C compared to conventional farming, regardless of soil types. Additionally, Org-NA had the highest dehydrogenase activity among treatments, whereas no difference was found between Org-A, Con-A and Con-NA and had the highest species richness (Chao 1) and diversity (Phyrogenetic diversity). Particularly, Chao 1 and Phyrogenetic diversity were increased in organic plots by 12% and 20%, compared with conventional plots, respectively. Also, regardless of agricultural management and soil types, Proteobacteria was the most abundant bacterial phylum, accounting for 21.9-25.9% of the bacterial 16S rRNAs. The relative abundance of putative copiotroph such as Firmicutes was highest in Org-NA plot by 21.0%, as follows Con-NA (13.1%), Con-A (6.7%) and Org-A (5.1%.), respectively and those of putative oligotrophs such as Acidobacteria and Planctomycetes were higher in Con-A than those in the other plots. Furthermore, LEfSe indicated that organic system enhanced the abundance of Fumicutes, while conventional system increased the abundance of Acidobacteria, especially in Non-andisols. Correlation analysis showed that total organic carbon (TOC) and nutrient levels (e.g. available P and exchangeable K) were significantly correlated to the structure of the microbial community and microbial activity. Overall, our results showed that the continuous organic farming systems without chemical materials, as well as the soil types made by long-term environmental factors might influence on soil properties and increase microbial abundances and diversity.

Fermentative characteristics of yogurt using lactic acid bacteria isolated from Korean traditional fermented food (전통 발효 식품에서 분리한 유산균을 이용한 yogurt 발효특성)

  • Park, Na-Young;Lee, Shin-Ho
    • Food Science and Preservation
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    • v.24 no.5
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    • pp.707-713
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    • 2017
  • The objective of this study was to select yogurt starter from Korean traditional fermented foods. The 2 strains (KM24, KM32) among 50 strains of isolated lactic acid bacteria selected as starter based on milk clotting ability, antimicrobial activity against various pathogens, tolerance in artificial gastric and bile juice and growth in 10 % skimmed milk. The strains were identified as Lacobacillus plantarum (KM32) and Pediococcus pentosacesus (KM24) by 16S rRNA gene sequencing. Viable cell number of yogurt fermented with mixed strains (KM24 and KM32) was 9.66 log CFU/mL after fermentation for 48 h and maintained $10^9CFU/mL$ during fermentation for 72 h at $37^{\circ}C$. The pH and titratable acidity of mixed cultured yogurt were 4.25% and 0.83% after fermentation for 48 h at $37^{\circ}C$, respectively. The physico-chemical characteristics of mixed cultured yogurt after fermentation for 48 h were $38.45{\mu}g/mL$ (polyphenol content), 48.57% (DPPH radical scavenging activity) and 465.40 cp (viscosity), respectively. The mixed cultured yogurt maintained $10^9CFU/mL$ of lactic acid bacteria during storage 10 days at $4^{\circ}C$. The viable cell number of yogurt prepared with mixed culture(KM32+KM24) maintained higher and than that of control (L. casei) during storage. These results indicated the potential use of selected strains (KM32+KM24) isolated from kimchi as a yogurt starter with strong acid tolerance and probiotics properties.

Biogeochemical Remediation of Cr(VI)-Contaminated Groundwater using MMPH-0 (Enterobacter aerogenes) (MMPH-0 (Enterobacter aerogenes)에 의한 6가 크롬 오염 지하수의 생지화학적 정화)

  • Seo, Hyun-Hee;Rhee, Sung-Keun;Kim, Kang-Joo;Park, Eun-Gyu;Kim, Yeong-Kyoo;Chon, Chul-Min;Moon, Ji-Won;Roh, Yul
    • Economic and Environmental Geology
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    • v.45 no.2
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    • pp.105-119
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    • 2012
  • Indigenous bacteria isolated from contaminated sites play important roles to remediate contaminated groundwater. Chromium has the most stable oxidation states. Cr(VI) is toxic, carcinogenic, and mobile, but Cr(III) is less toxic and immobile. In this study, indigenous microorganism (MMPH-0) was enriched from Cr(VI) contaminated groundwater, and identified by 16S rRNA gene analysis. Using MMPH-0, the effect of stimulating with e-donors (glucose, lactate, acetate, and no e-donor control), respiration conditions, biomass, tolerance, and geochemical changes on Cr(VI) reduction were investigated in batch experiments for 4 weeks. The changes of Cr(VI) concentration and geochemical conditions were monitored using UV-vis-spectrophotometer and Eh-pH meter. And the morphological and chemical characteristics of MMPH-0 and precipitates in the effluents were characterized by TEM-EDS and SEM-EDS analyses. MMPH-0 (Enterobacter aerogenes) was able to tolerate up to 2000 mg/L Cr(VI) and reduce Cr(VI) under aerobic and anaerobic conditions. MMPH-0 performed faster and higher efficiency of Cr(VI) reduction with electron donors (over 70% after 1 week with e-donor, 10-20% after 4 weeks without e-donor). The changes of Eh-pH in effluents showing the tendency from oxidizing to reducing condition and a bit of acidic change in pH due to microbial oxidation of organic matters donating electrons and protons suggested the roles of MMPH-0 on Cr(VI) in the contaminated water catalyzing to transit geochemical stable zone for more stable $Cr(OH)_3$ or Cr(III) precipitates. TEM/SEM-EDS analyses of MMPH-0 and precipitates indicate direct and indirect Cr(VI) reduction: extracellular polymers capturing Cr component outside cells. These results suggested diverse indigenous bacteria and their biogeochemical reactions might enhance more effective and feasible remediation technology of redox sensitive heavy metals in metal-contaminated in groundwater.

Optimization of the Indole-3-Acetic Acid Production Medium of Pantoea agglomerans SRCM 119864 using Response Surface Methodology (반응표면분석법을 활용한 Pantoea agglomerans SRCM 119864의 Indole-3-acetic acid 생산 배지 최적화)

  • Ho Jin, Jeong;Gwangsu, Ha;Su Ji, Jeong;Myeong Seon, Ryu;JinWon, Kim;Do-Youn, Jeong;Hee-Jong, Yang
    • Journal of Life Science
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    • v.32 no.11
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    • pp.872-881
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    • 2022
  • In this study, we optimized the composition of the indole-3-acetic acid (IAA) production medium using response surface methodology on Pantoea agglomerans SRCM 119864 isolated from soil. IAA-producing P. aglomerans SRCM 119864 was identified by 16S rRNA gene sequencing. There are 11 intermediate components known to affect IAA production, hence the effect of each component on IAA production was investigated using a Plackett-Burman design (PBD). Based on the PBD, sucrose, tryptone, and sodium chloride were selected as the main factors that enhanced the IAA production at optimal L-tryptophan concentration. The predicted maximum IAA production (64.34 mg/l) was obtained for a concentration of sucrose of 13.38 g/l, of tryptone of 18.34 g/l, of sodium chloride of 9.71 g/l, and of L-tryptophan of 6.25 g/l using a the hybrid design experimental model. In the experiment, the nutrient broth medium supplemented with 0.1% L-tryptophan as the basal medium produced 45.24 mg/l of IAA, whereas the optimized medium produced 65.40 mg/l of IAA, resulting in a 44.56% increase in efficiency. It was confirmed that the IAA production of the designed optimal composition medium was very similar to the predicted IAA production. The statistical significance and suitability of the experimental model were verified through analysis of variance (ANOVA). Therefore, in this study, we determined the optimal growth medium concentration for the maximum production of IAA, which can contribute to sustainable agriculture and increase crop yield.