• Title/Summary/Keyword: 16S-rRNA

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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).

Isolation and Characterization of 𝛽-Glucosidase-Producing Yeast, Rhodotorula sp. GYP-1 (𝛽-Glucosidase 생성 효모 Rhodotorula sp. GYP-1의 분리 및 특성)

  • Hyun-Soo Roh;Min-Young Kwon;Sol-Bi Kim;Jae-Eun Cho;Song-Ih Han
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.5
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    • pp.1126-1135
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    • 2023
  • Nine microbial strains were isolated from the byproduct of ginseng processing and field of ginseng cultivation. Two strains among them were confirmed. Phylogenetic analysis of these 𝛽-Glucosidase strains confirmed that strain GYP-1 belongs to the Rhodotorula and strain GYP-3-3 belong to genus Brachybacterium. Rhodotorula sp. GYP-1 was finally selected due to its high biomass production. The 𝛽-Glucosidase activity of Rhodotorula sp. GPY-1 was assessed at 30 ℃, and Higher than 70% of the enzyme activity was maintained at the temperature range of 20-40℃. Although the optimum pH for the highest enzyme activity was pH 5.0, the enzyme was stable throughout the pH range of 5.0-8.0. In addition, Rhodotorula sp. demonstrated antifungal activity against the ginseng root rot disease caused by Botrytis.

Characterization of Photobacterium sp. YW2207 isolated from rainbow trout (Oncorhynchus mykiss) raised in a fresh water farm in South Korea (국내 양식 무지개송어(Oncorhynchus mykiss)에서 분리된 Photobacterium sp. YW2207의 특성)

  • Hyunwoo Kim;Eunsup Lee;Sung Jun Lee;Haneul Kim;So-Ra Han;Tae-Jin Oh;Myoung Sug Kim;Soo-Jin Kim;Se Ryun Kwon
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.251-261
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    • 2023
  • Photobacterium sp. YW2207 was isolated from rainbow trout raised in a fish farm located in Yeongwol-gun, Gangwon Province, South Korea. Based on 16S rRNA sequence analysis and phylogenetic analysis, it was confirmed that Photobacterium sp. YW2207 showed 100% similarity with Photobacterium piscicola and Photobacterium phosphoreum, and 94.6% similarity with P. damselae subsp. damselae. Biochemical analysis revealed that Photobacterium sp. YW2207 is a Gram-negative, motile bacterium with a cell size of 1.5~3×3~5 ㎛. The bacteria were cultured on nutrient agar, brain heart infusion agar, Muller-Hinton agar, tryptic soy agar, and thiosulfate citrate bile sucrose agar with NaCl concentrations ranging from 0 to 2.5%. The API50CHE and API20E tests indicated lower utilization capabilities compared to the P. damselae strains provided in the API database. Furthermore, unlike most Photobacterium species, Photobacterium sp. YW2207 presented negative for catalase test. Results from the flow cytometric measurement indicated that Photobacterium sp. YW2207 exhibited a more diverse distribution of cell sizes and had larger cell sizes compared with P. damselae subsp. damselae. Minimum inhibitory concentration tests showed that Photobacterium sp. YW2207 had low susceptibility to β-Lactam and aminoglycoside antibiotics, while having high susceptibility to tetracycline, doxycycline, and quinolone antibiotics. Pathogenicity on rainbow trout revealed that an immersion of 1×105 CFU/ml did not cause mortality or clinical symptoms.

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.