• Title/Summary/Keyword: Gut bacteria

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Novel Alkali-Tolerant GH10 Endo-${\beta}$-1,4-Xylanase with Broad Substrate Specificity from Microbacterium trichothecenolyticum HY-17, a Gut Bacterium of the Mole Cricket Gryllotalpa orientalis

  • Kim, Do Young;Shin, Dong-Ha;Jung, Sora;Kim, Hyangmi;Lee, Jong Suk;Cho, Han-Young;Bae, Kyung Sook;Sung, Chang-Keun;Rhee, Young Ha;Son, Kwang-Hee;Park, Ho-Yong
    • Journal of Microbiology and Biotechnology
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    • v.24 no.7
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    • pp.943-953
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    • 2014
  • The XylH gene (1,167-bp) encoding a novel hemicellulase (41,584 Da) was identified from the genome of Microbacterium trichothecenolyticum HY-17, a gastrointestinal bacterium of Gryllotalpa orientalis. The enzyme consisted of a single catalytic domain, which is 74% identical to that of an endo-${\beta}$-1,4-xylanase (GH10) from Isoptericola variabilis 225. Unlike other endo-${\beta}$-1,4-xylanases from invertebrate-symbiotic bacteria, rXylH was an alkali-tolerant multifunctional enzyme possessing endo-${\beta}$-1,4-xylanase activity together with ${\beta}$-1,3/${\beta}$-1,4-glucanase activity, which exhibited its highest xylanolytic activity at pH 9.0 and 60oC, and was relatively stable within a broad pH range of 5.0-10.0. The susceptibilities of different xylosebased polysaccharides to the XylH were assessed to be as follows: oat spelts xylan > beechwood xylan > birchwood xylan > wheat arabinoxylan. rXylH was also able to readily cleave p-nitrophenyl (pNP) cellobioside and pNP-xylopyranoside, but did not hydrolyze other pNP-sugar derivatives, xylobiose, or hexose-based materials. Enzymatic hydrolysis of birchwood xylan resulted in the product composition of xylobiose (71.2%) and xylotriose (28.8%) as end products.

Pomegranate (Punica granatum L.) Peel Extract Inhibits Quorum Sensing and Biofilm Formation Potential in Yersinia enterocolitica (석류 껍질추출물이 식중독균 여시니아 엔테로콜리티카의 쿼럼센싱과 바이오필름 형성능 억제)

  • Oh, Soo Kyung;Chang, Hyun Joo;Chun, Hyang Sook;Kim, Hyun Jin;Lee, Nari
    • Microbiology and Biotechnology Letters
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    • v.43 no.4
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    • pp.357-366
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    • 2015
  • Quorum sensing (QS) is involved in the process of cell-to-cell communication and as a gene regulatory mechanism, which has been implicated in bacterial pathogenicity. Bacteria use this QS system to control a variety of physiological processes. In this study, pomegranate (Punica granatum L.) peel extract (PPE) was first screened for its ability to inhibit QS in bio-reporter strains (Chromobacterium violaceum and C. violaceum CV026). Next, the ability of PPE to inhibit swimming motility and biofilm formation was examined in Y. enterocolitica. Additionally, changes in the expression of specific genes involved in the synthesis of the N-acylhomoserine lactones (AHLs; yenI and yenR) and in the flagellar regulon (fliA, fleB and flhDC) were evaluated by reverse transcription (RT)-PCR. The results show that PPE specifically inhibited and reduced QS-controlled violacein production by 78.5% in C. violaceum CV026, and decreased QS-associated biofilm formation and swimming motility in Y. enterocolitica without significantly affecting bacterial growth. These inhibitory effects were also associated with the down-regulation of gene expression involved in the synthesis of AHLs and in motility. Our results suggest that PPE could be a potential therapeutic agent to prevent enteropathogens in humans, as well as highlight the need to further investigate the in vivo properties of PPE for clinical applications.

Expression of Toll-like Receptors, Pro-, and Anti-inflammatory Cytokines in Relation to Gut Microbiota in Irritable Bowel Syndrome: The Evidence for Its Micro-organic Basis

  • Shukla, Ratnakar;Ghoshal, Ujjala;Ranjan, Prabhat;Ghoshal, Uday C
    • Journal of Neurogastroenterology and Motility
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    • v.24 no.4
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    • pp.628-642
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    • 2018
  • Background/Aims A Subset of patients with irritable bowel syndrome (IBS) may have mild inflammation due to immune activation. Toll-like receptors (TLRs) and cytokines may cause intestinal inflammation. We studied their expression in relation to gut microbiota. Methods Expression of TLRs and cytokines was assessed in 47 IBS patients (Rome III) and 25 controls using quantitative real-time polymerase chain reaction. Immunohistochemistry was further performed to confirm the expression of TLR-4 and TLR-5. Results Of 47 patients with IBS, 20 had constipation (IBS-C), 20 diarrhea (IBS-D), and 7 unclassified (IBS-U). The mRNA levels of TLR-4 and TLR-5 were up-regulated in IBS patients than controls (P = 0.013 and P < 0.001, respectively). Expression of TLR-4 and TLR-5 at protein level was 4.2-folds and 6.6-folds higher in IBS-D than controls. The mRNA levels of IL-6 (P = 0.003), C-X-C motif chemokine ligand 11 (CXCL-11) (P < 0.001) and C-X-C motif chemokine receptor 3 (CXCR-3) (P < 0.001) were higher among IBS patients than controls. Expression of IL-6 (P = 0.002), CXCL-11 (P < 0.001), and CXCR-3 (P < 0.001) were up-regulated and IL-10 (P = 0.012) was down-regulated in IBS-D patients than controls. Positive correlation was seen between TLR-4 and IL-6 (P = 0.043), CXCR-3, and CXCL-11 (P = 0.047), and IL-6 and CXCR-3 (P = 0.003). Stool frequency per week showed positive correlation with mRNA levels of TLR-4 (P = 0.016) and CXCR-3 (P = 0.005), but inversely correlated with IL-10 (P = 0.002). Copy number of Lactobacillus (P = 0.045) and Bifidobacterium (P = 0.011) showed correlation with IL-10 in IBS-C, while Gram-positive (P = 0.031) and Gram-negative bacteria (P = 0.010) showed correlation with CXCL-11 in IBS-D patients. Conclusions Altered immune activation in response to dysbiotic microbiota may promote intestinal inflammation in a subset of patients with IBS.

Production of γ-amino Butyric Acid by Lactic Acid Bacteria in Skim Milk (탈지방우유에서 가바생성 유산균 배양을 통한 가바생성 연구)

  • Cha, Jin Young;Kim, Young Rok;Beck, Bo Ram;Park, Ji Hun;Hwang, Cher Won;Do, Hyung Ki
    • Journal of Life Science
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    • v.28 no.2
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    • pp.223-228
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    • 2018
  • Lactic acid bacteria were isolated from a variety of fermented seafoods and sea creatures from the East Sea Rim, Korea and were screened for ${\gamma}-amino$ butyric acid-producing (GABA) activity. Through a 16S rRNA sequence analysis, the bacteria of interest, which were GABA-positive on the thin-layer chromatography analysis, were recognized as three isolates of Lactobacillus (Lb.) brevis and one isolate of Lactococcus (Lc.) lactis. Lb. brevis FSFL0004 and FSFL0005 were isolated from fermented anglerfish and Lb. brevis FSFL0036 was derived from salted cutlass fish. The Lc. lactis strain FGL0007 was isolated from the gut of a brown sole flounder. According to HPLC analysis, the GABA contents produced by FSFL0004, FSFL0005, FSFL0036 and FGL0007 were equivalent to $10,754.37{\mu}g/ml$, $13,082.79{\mu}g/ml$, $12,290.19{\mu}g/ml$, and $45.07{\mu}g/ml$ respectively in 1% monosodium glutamate-supplemented methionyl-tRNA synthetase (MRS) broth. The four strains were inoculated in skim milk with 1% monosodium glutamate to commercialize the strains as starter cultures for GABA-enriched dairy products, and TLC results displayed the production of ${\gamma}-amino$ butyric acid by all four strains in the adaptation media. Lc. lactis FGL0007 demonstrated the greatest GABA production ($431.42{\mu}g/ml$) by HPLC analysis. The GABA production by lactic acid bacteria strains in the skim milk demonstrated in the present study may be helpful for the production of GABA-enriched dairy products.

Physiochemical Treatment of Feed and Utilization of Feed Additives to Control Salmonella in Poultry (가금의 살모넬라 제어를 위한 사료의 이화학적 처리와 사료첨가제의 활용)

  • Kim, Ji-Hyuk;Kim, Hack-Youn;Kim, Bong-Ki;Kim, Gye-Woong
    • Korean Journal of Poultry Science
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    • v.45 no.1
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    • pp.1-15
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    • 2018
  • Salmonella infections in livestock industry cause various problems such as worsening animal welfare and productivity, damaging consumer confidence in the food safety of animal products. Chicken meat and eggs are known as major source of pathogen causing human foodborne infections. Therefore food safety concerns have prompted the poultry producers and governments to introduce the strategy and regulation to control these pathogens. Salmonella can persist for long periods of time in a wide range of spaces including feed bin, feed processing facilities, poultry farm, slaughterhouse, processing plants, etc. For the effective and constant Salmonella control, combination of pre-harvest, harvest and post-harvest measures should be considered comprehensively. The control measures would be most effective at farm level where the contamination initiates. Transmission of pathogen from feed origin to the live poultry and finally to the products was proven already. To control bacteria in the feed ingredients and formula feed, thermal processing, irradiation or chemical treatment may be applied. Chemical treatments to inhibit Salmonella in the feed involve the use of products containing organic acids, formaldehyde, or a combination of such compounds. However, recontamination which might occur during storage and transport process and/or by other various factors should always be under control and eliminated. Feed additives used to control Salmonella in birds' gastrointestinal track can be of various types, including prebiotics, probiotics, organic acids and bacteriophages. Although their mode of action varies, they ultimately inhibit the colonization of Salmonella in the gut and improve the performance of birds. This review describes the strategies that could be adapted to the management of feedstuffs and the use of feed additives in pre-harvest stage to control Salmonella contamination in poultry farming.

Bacteroides fragilis Toxin Induces IL-8 Secretion in HT29/C1 Cells through Disruption of E-cadherin Junctions

  • Hwang, Soonjae;Gwon, Sun-Yeong;Kim, Myung Sook;Lee, Seunghyung;Rhee, Ki-Jong
    • IMMUNE NETWORK
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    • v.13 no.5
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    • pp.213-217
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    • 2013
  • Enterotoxigenic Bacteroides fragilis (ETBF) is a human gut commensal bacteria that causes inflammatory diarrhea and colitis. ETBF also promotes colorectal tumorigenesis in the Min mouse model. The key virulence factor is a secreted metalloprotease called B. fragilis toxin (BFT). BFT induces E-cadherin cleavage, cell rounding, activation of the ${\beta}$-catenin pathway and secretion of IL-8 in colonic epithelial cells. However, the precise mechanism by which these processes occur and how these processes are interrelated is still unclear. E-cadherin form homophilic interactions which tethers adjacent cells. Loss of E-cadherin results in detachment of adjacent cells. Prior studies have suggested that BFT induces IL-8 expression by inducing E-cadherin cleavage; cells that do not express E-cadherin do not secrete IL-8 in response to BFT. In the current study, we found that HT29/C1cells treated with dilute trypsin solution induced E-cadherin degradation and IL-8 secretion, consistent with the hypothesis that E-cadherin cleavage causes IL-8 secretion. However, physical damage to the cell monolayer did not induce IL-8 secretion. We also show that EDTA-mediated disruption of E-cadherin interactions without E-cadherin degradation was sufficient to induce IL-8 secretion. Finally, we determined that HT29/C1 cells treated with LiCl (${\beta}$-catenin activator) induced IL-8 secretion in a dose-dependent and time-dependent manner. Taken together, our results suggest that BFT induced IL-8 secretion may occur by the following process: E-cadherin cleavage, disruption of cellular interactions, activation of the ${\beta}$-catenin pathway and IL-8 expression. However, we further propose that E-cadherin cleavage per se may not be required for BFT induced IL-8 secretion.

Effects of probiotic supplement (Bacillus subtilis and Lactobacillus acidophilus) on feed efficiency, growth performance, and microbial population of weaning rabbits

  • Phuoc, Thanh Lam;Jamikorn, Uttra
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.198-205
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    • 2017
  • Objective: This study aimed to investigate the effects of single or/and double strains of probiotic supplement on feed efficiency, growth performance, and microbial population in distal gastrointestinal tract (GIT) of weaning rabbits. Methods: Sixty-four weaning (28 days old) New Zealand White rabbits were randomly distributed into four groups with treatments including: basal diet without probiotic supplement (control) or supplemented as follows: $1{\times}10^6cfu/g$ B. subtilis (BS group), $1{\times}10^7cfu/g$ L. acidophilus (LA group), or $0.5{\times}10^6cfu/g$ B. subtilis plus $0.5{\times}10^7cfu/g$ L. acidophilus (BL group). During the research, the male and female rabbits were fed separately. Body weight of the rabbits was recorded at 28, 42, and 70 d of age. Results: There was an increase (p<0.05) in body weight gain for the LA group at 42 d. Rabbits fed BL responsed with a greater growth (p<0.05) and better feed conversion ratio (p<0.05) than those fed with no probiotic. Digestibility coefficients of dry matter, organic matter, crude protein, neutral detergent fiber, and gross energy were higher (p<0.05) in LA and BL groups than those in the control group. Male rabbits had higher (p<0.05) Bacilli spp. and Coliformis spp. in the ileum than female rabbits. Rabbits supplemented with BS had greater (p<0.05) numbers of bacilli in all intestinal segments than those receiving no probiotic, whereas intestinal Lactobacilli populations were greater (p<0.001) in the LA and BL diets compared to control. Average intestinal coliform populations were lowest (p<0.05) in the rabbits supplemented with LA as compared to those fed the control and BS. Conclusion: Supplementation of L. acidophilus alone or in combination with B. subtilis at a half of dose could enhance number of gut beneficial bacteria populations, nutrient digestibility, cecal fermentation, feed efficiency, and growth performance, but rabbits receiving only B. subtilis alone were not different from the controls without probiotic.

Effects of Biologically Active Substances in Natural Products on the Hepatic Detoxication Mechanism (천연물중의 생리활성성분이 간해독기구에 미치는 영향)

  • 권정숙
    • Journal of Nutrition and Health
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    • v.27 no.4
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    • pp.347-355
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    • 1994
  • Indolo[3,2-b]carbazole(ICZ) is a potent Ah receptor agonist with biological activities similar in several respects to those of the potent environmental toxin, TCDD. ICZ is produced during the oilgomerization of indole-3-carbinol(I3C), a breakdown product of the glucobrassicin present in food plants of the Brassica genus. In the present study we examined ICZ levels in tissues and excreta of rats treated with I3C or dietary cabbage of established glucobrasicin content, and in feces of conventional and germfree rats fed on a basal diet, and of humans. We also examined the levels of cytochrome P4501A1 induction, as determined by the ethoxyresorufin ο-deethylase assay, in tissues of animals that received cabbage-supplemented diets, or which were treated with purified I3C or ICZ. Our findings indicated that incorporation of either homogenized or whole freeze-dried cabbage in the feed led to large increases(16-60 fold) in the levels of ICZ in the feces and lower gastrointestinal tract of rats. We observed that whereas ICZ is readily detectable at about the same levels(2.00$\pm$0.50 ppb) in the feces of conventional rats fed on a purified diet and in human feces, levels of ICZ in the feces of germfree animals fed on the basal diet were at the limits of detection(0.40$\pm$0.20 ppb), indication that gut bacteria are important for the production of ICZ from essential dietary constituents in the basal diet. We showed that in contrast to the near 7000-fold difference in CYP1A1 inducing potencies of ICZ and TCDD in cells in culture, their inducing potencies differ by only about an order of magnitude in rats. Nonetheless, the levels of ICZ remaining in livers twenty hours after I3C treatment appear too low to account for the induced activity. This result indicates that ICZ may be rapidly cleared from the liver or that substances other than, or in addition to, ICZ be responsible for the enzyme-inducing activity of orally administered I3C or its precursors.

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Assay of ${\beta}$-Glucosidase Activity of Bifidobacteria and the Hydrolysis of Isoflavone Glycosides by Bifidobacterium sp. Int-57 in Soymilk Fermentation

  • Jeon, Ki-Suk;Ji, Geun-Eog;Hwang, In-Kyeong
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.8-13
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    • 2002
  • The isoflavone glycosides are hydrolyzed by ${\beta}$-glucosidase from gut microbes to the bioactive aglycones. However, the specific bacteria from the human intestinal tract that are involved in the metabolism of these compounds are not known. This study was undertaken to develop a fermented soymilk which converts isoflavones to the more bioactive aglycones form using a Bifidobacterium strain. The ${\beta}$-glucosidase activity of 15 Bifidobacterium strains were measured during cell growth. Among them, Bifidobacterium sp. Int-57 was selected for this study, because it has the highest ${\beta}$-glucosidase activity. Growth, acid development, ${\beta}$-glucosidase activity, and the hydrolysis of daidzin and genistin were investigated in four soymilks inoculated with Bifidobacterium sp. Int-57. After 12 h of fermentation, the counts of viable Bifidobacterium sp. Int-57 in all the soymilks reached a level of more than $10^8$ cfu/ml, which was then maintained. The pH of soymilks started to decrease rapidly after 6 h of fermentation and leveled off after 18 h. The titratable acidity of BL# 1 soymilk, BL#2 soymilk, and JP#l soymilk increased from 0.18 to 1.21, 1.15, and $1.08\%$ over the fermentation period, respectively. After 24 h of fermentation, the $\beta$-glucosidase activity in BL#1 soymilk, BL#2 soymilk, JP#l soymilk, and JP#2 soymilk increased to 59.528, 40.643, 70.844, and 56.962 mU/ml, respectively. The isoflavone glycosides, daidzin and genistin, in soymilks were hydrolyzed completely in the relatively short fermentation time of 18 h. These results show that Bifidobacterium sp. Int-57 can be used as a potential starter culture for developing fermented soymilk which has completely hydrolyzed isoflavone glycosides.

Value of spray-dried egg in pig nursery diets

  • Song, Minho;Kim, Sheena;Kim, Younghwa;Park, Juncheol;Kim, Younghoon
    • Korean Journal of Agricultural Science
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    • v.42 no.3
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    • pp.207-213
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    • 2015
  • High-quality protein ingredients have been used in nursery diets, in spite of expensive ingredients, to minimize nutritional deficiency and disease problems. Recent dramatic increases in prices of protein products for nursery diets have exacerbated the challenge. Spray-dried egg may be a part of the solutions. Therefore, this review describes the value of spray-dried egg in nursery diets as a high-quality protein source. Spray-dried egg is egg by-product and is produced by only eggs without shell that are below the USDA Grade B standards. Spray-dried egg is an excellent nutrient source: 1) highly digestible, 2) excellent balance of amino acids, 3) rich content of fat, and 4) high metabolizable energy. These can be attributed to growth of nursery pigs. Beyond the provision of bioavailable nutrients, spray-dried egg also may provide specific physiological benefits. Spray-dried egg contains 1) immunoglobulin antibodies (IgY: IgG in egg yolk) that may attach to intestinal pathogens and excrete them and 2) lysozymes antimicrobial protein that can damage bacteria cell wall. Thereby feeding spray-dried egg may reduce concentration of intestinal pathogen and thus improve potential gut health or enteric disease resistance in nursery pigs. This is important for physiologically immature weaned pigs. Based on these benefits, spray-dried egg is believed to have the same benefits as spray-dried plasma protein and milk products in diets for nursery pigs. Therefore, it is suggested that spray-dried egg has a great potential as a valuable protein source in nursery diets.