• Title/Summary/Keyword: 16S-rRNA

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Correlation analysis of muscle amino acid deposition and gut microbiota profile of broilers reared at different ambient temperatures

  • Yang, Yuting;Gao, Huan;Li, Xing;Cao, Zhenhui;Li, Meiquan;Liu, Jianping;Qiao, Yingying;Ma, Li;Zhao, Zhiyong;Pan, Hongbin
    • Animal Bioscience
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    • v.34 no.1
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    • pp.93-101
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    • 2021
  • Objective: Temperature could influence protein and amino acid deposition as well as gut microbiota profile and composition. However, the specific effects of ambient temperature on amino acids deposition and gut microbiota composition remain insufficiently understood. Methods: A total of 300 one-day-old Avian broilers were randomly divided into three groups and reared at high, medium, and low temperature (HT, MT, and LT), respectively. Breast muscle and fecal samples were collected for amino acid composition analysis and 16S rRNA gene sequence analysis. Results: Our data showed that compared to the MT group, there was a decrease of muscle leucine and tyrosine (p<0.05), as well as an increase of methionine in the HT group (p<0.05) and a decrease of serine in the LT group. Examination of microbiota shift revealed that at genus level, the relative abundance of Turicibacter and Parabacteroides was increased in the HT group (p<0.05) and that the relative abundances of Pandoraea, Achromobacter, Prevotella, Brevundimonas, and Stenotrophomonas in the LT group were higher than those in the MT group (p<0.05). In addition, there were substantial correlations between microbes and amino acids. In the HT group. Turicibacter was negatively correlated with aspartic acid and tyrosine, whereas Parabacteroides was positively correlated with methionine (p<0.05). In the LT group, there were multiple positive correlations between Achromobacter and arginine, isoleucine or tyrosine; between Prevotella and cysteine or phenylalanine; between Brevundimonas and cysteine; and between Stenotrophomonas and cysteine as well as a negative correlation between Stenotrophomonas and serine. Conclusion: Our findings demonstrated that amino acid content of breast muscle and intestinal microbiota profile was affected by different ambient temperatures. Under heat exposure, augmented abundance of Parabacteroides was correlated with elevated methionine. Low temperature treatment may affect muscle tyrosine content through the regulation of Achromobacter.

Effect of Korea red ginseng on nonalcoholic fatty liver disease: an association of gut microbiota with liver function

  • Hong, Ji Taek;Lee, Min-Jung;Yoon, Sang Jun;Shin, Seok Pyo;Bang, Chang Seok;Baik, Gwang Ho;Kim, Dong Joon;Youn, Gi Soo;Shin, Min Jea;Ham, Young Lim;Suk, Ki Tae;Kim, Bong-Soo
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.316-324
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    • 2021
  • Background: Korea Red Ginseng (KRG) has been used as remedies with hepato-protective effects in liver-related condition. Microbiota related gut-liver axis plays key roles in the pathogenesis of chronic liver disease. We evaluated the effect of KRG on gut-liver axis in patients with nonalcoholic statohepatitis by the modulation of gut-microbiota. Methods: A total of 94 patients (KRG: 45 and placebo: 49) were prospectively randomized to receive KRG (2,000 mg/day, ginsenoside Rg1+Rb1+Rg3 4.5mg/g) or placebo during 30 days. Liver function test, cytokeraton 18, and fatigue score were measured. Gut microbiota was analyzed by MiSeq systems based on 16S rRNA genes. Results: In KRG group, the mean levels (before vs. after) of aspartate aminotransferase (53 ± 19 vs. 45 ± 23 IU/L), alanine aminotransferase (75 ± 40 vs. 64 ± 39 IU/L) and fatigue score (33 ± 13 vs. 26 ± 13) were improved (p < 0.05). In placebo group, only fatigue score (34 ± 13 vs. 31 ± 15) was ameliorated (p < 0.05). The changes of phyla were not statistically significant on both groups. In KRG group, increased abundance of Lactobacillus was related with improved alanine aminotransferase level and increased abundance of Clostridium and Intestinibacter was associated with no improvement after KRG supplementation. In placebo group, increased abundance of Lachnospiraceae could be related with aggravation of liver enzyme (p < 0.05). Conclusion: KRG effectively improved liver enzymes and fatigue score by modulating gut-microbiota in patients with fatty liver disease. Further studies are needed to understand the mechanism of improvement of nonalcoholic steatohepatitis. ClnicalTrials.gov: NCT03945123 (www.ClinicalTrials.gov).

Optimization of Medium to Improve Protease Production Using Response Surface Methodology by Bacillus amyloliquefaciens SRCM115785 (반응표면분석법을 이용한 Bacillus amyloliquefaciens SRCM115785의 protease 활성증가를 위한 배지 최적화)

  • Yang, Hee Gun;Ha, Gwangsu;Ryu, Myeong Seon;Park, Se Won;Jeong, Ho Jin;Yang, Hee-Jong;Jeong, Do-Youn
    • Journal of Life Science
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    • v.31 no.8
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    • pp.761-770
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    • 2021
  • In this study, the optimal medium composition for enhancing protease production was established by the Bacillus strain isolated from Makgeolli, a traditional fermented food, using the response surface methodology. B. amyloliquefaciens SRCM115785 was selected as the protease producer by productivity analysis and identified by 16S rRNA gene sequencing. Plackett-Burman design (PBD) was introduced to analyze the effect of each component on protease production among the 11 selected medium components. As a result, glucose, yeast extract, and beef extract were finally selected as factors for enhancing protease production. Central composite design (CCD) analysis was designed as a method to determine the optimal concentration of each component for protease production and the concentration of each medium composition for maximum protease production was predicted to glucose 6.75 g/l, yeast extract 12.42 g/l and beef extract 17.48 g/l. The suitability of the experimental model was proved using ANOVA analysis and as a result of quantitative analysis to prove this, the amount of increase was 230.47% compared to the LB medium used as a control. Through this study, the optimization of medium composition for enhancing protease production was established, and based on this, it is expected that it can be efficient use of protease as an industrial enzyme.

Biochemical properties and gluten degradation of Lactobacillus paracasei strain GLU70 isolated from salted seafood (젓갈에서 분리한 락토바실러스 파라카제이 GLU70 균주의 생화학적 특성 및 글루텐 분해능)

  • Park, Hyein;Yoon, Seul Gi;Jang, Junho;Byun, Ji Young;Yoon, Bok Kun
    • Korean Journal of Food Science and Technology
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    • v.54 no.2
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    • pp.203-208
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    • 2022
  • Gluten is an insoluble protein present in cereals such as wheat. Gluten consumed through food is not digested and accumulates in the body; this has been linked to digestive discomfort, irritation, and various digestive disorders, including intestinal inflammation. In this study, the Lactobacillus paracasei strain GLU70, which exhibits a glutendegrading ability, was isolated from salted seafood. At a pH of 3.0, GLU70 showed a survival rate of approximately 84%, and at 0.3% oxgall, it showed a survival rate of approximately 53%. When the culture supernatant collected after 12 h of incubation was added to flour dough, approximately 50% gluten degradation was observed. Moreover, among several probiotic isolates exhibiting proteolytic activity selected to assess the gluten-degrading ability, GLU70 showed superior results regardless of the dough fermentation temperature. Although further research is required, GLU70 is expected to be of value in manufacturing gluten-reduced products and the food industry as an ingredient or additive.

Simotang Alleviates the Gastrointestinal Side Effects of Chemotherapy by Altering Gut Microbiota

  • Deng, Lijing;Zhou, Xingyi;Lan, Zhifang;Tang, Kairui;Zhu, Xiaoxu;Mo, Xiaowei;Zhao, Zongyao;Zhao, Zhiqiang;Wu, Mansi
    • Journal of Microbiology and Biotechnology
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    • v.32 no.4
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    • pp.405-418
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    • 2022
  • Simotang oral liquid (SMT) is a traditional Chinese medicine (TCM) consisting of four natural plants and is used to alleviate gastrointestinal side effects after chemotherapy and functional dyspepsia (FD). However, the mechanism by which SMT helps cure these gastrointestinal diseases is still unknown. Here, we discovered that SMT could alleviate gastrointestinal side effects after chemotherapy by altering gut microbiota. C57BL/6J mice were treated with cisplatin (DDP) and SMT, and biological samples were collected. Pathological changes in the small intestine were observed, and the intestinal injury score was assessed. The expression levels of the inflammatory factors IL-1β and IL-6 and the adhesive factors Occludin and ZO-1 in mouse blood or small intestine tissue were also detected. Moreover, the gut microbiota was analyzed by high-throughput sequencing of 16S rRNA amplicons. SMT was found to effectively reduce gastrointestinal mucositis after DDP injection, which lowered inflammation and tightened the intestinal epithelial cells. Gut microbiota analysis showed that the abundance of the anti-inflammatory microbiota was downregulated and that the inflammatory microbiota was upregulated in DDP-treated mice. SMT upregulated anti-inflammatory and anticancer microbiota abundance, while the inflammatory microbiota was downregulated. An antibiotic cocktail (ABX) was also used to delete mice gut microbiota to test the importance of gut microbiota, and we found that SMT could not alleviate gastrointestinal mucositis after DDP injection, showing that gut microbiota might be an important mediator of SMT treatment. Our study provides evidence that SMT might moderate gastrointestinal mucositis after chemotherapy by altering gut microbiota.

Supplementation of Microalgae (Tetradesmus sp.) to Pre-Starter Diet for Broiler Chickens (초기 육계 사료 내 미세조류(Tetradesmus sp.) 첨가에 따른 장관환경 변화)

  • Rim, Chae Yun;Jung, Hui-su;An, Su Hyun;Joo, Sang Seok;Kim, Z-Hun;Kong, Changsu;Kim, Myunghoo
    • Korean Journal of Poultry Science
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    • v.49 no.2
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    • pp.125-137
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    • 2022
  • This study aimed to determine the effects of dietary microalgae (Tetradesmus sp. (TO)) on intestinal immunity and microbiota of pre-starter broilers. One hundred and twenty 1-day-old birds (Ross 308) were allocated to two dietary treatment groups with six blocks in a randomized complete block design. The two experimental diets consisted of a corn-soybean meal-based basal diet and a diet with 0.5% TO powder instead of cornstarch in the basal diet. After feeding the experimental diets for ten days, all birds' body weight and feed intake were measured, and representative eight birds were selected from each treatment group. Small intestinal lamina propria cells were isolated using flow cytometry to examine the frequency of immune cells. Cecal feces were harvested for 16s rRNA gut microbiota analysis and fecal IgA levels. Here, we found that 0.5% TO supplementation increased CD3+CD4+ T cells in the small intestine, but decreased CD3+CD8+ T cells in the small intestine. Gut microbial analysis showed that TO supplementation significantly increased the alpha diversity of the gut microbiome. Taxonomic analysis showed that TO treatment increased the abundance of Firmicutes and decreased that of Bacteroidetes at the phylum level. The distribution of Enterobacteriaceae containing many harmful bacteria at the family level, was lower in the TO group. In the LEfSe analysis, the TO group had a significantly enriched abundance of Agathobaculum at the genus level. Overall, results show that Tetradesmus sp. supplementation influences intestinal T-cell immunity and induces the expansion of beneficial gut microbes in pre-starter broiler chickens.

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.

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.

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).