Enterococcus faecium PNC01 isolated from the intestinal mucosa of chicken as an alternative for antibiotics to reduce feed conversion rate in broiler chickens

Abstract Background The development and utilization of probiotics had many environmental benefits for replacing antibiotics in animal production. Bacteria in the intestinal mucosa have better adhesion to the host intestinal epithelial cells compared to bacteria in the intestinal contents. In this study, lactic acid bacteria were isolated from the intestinal mucosa of broiler chickens and investigated as the substitution to antibiotic in broiler production. Results In addition to acid resistance, high temperature resistance, antimicrobial sensitivity tests, and intestinal epithelial cell adhesion, Enterococcus faecium PNC01 (E. faecium PNC01) was showed to be non-cytotoxic to epithelial cells. Draft genome sequence of E. faecium PNC01 predicted that it synthesized bacteriocin to perform probiotic functions and bacteriocin activity assay showed it inhibited Salmonella typhimurium from invading intestinal epithelial cells. Diet supplemented with E. faecium PNC01 increased the ileal villus height and crypt depth in broiler chickens, reduced the relative length of the cecum at day 21, and reduced the relative length of jejunum and ileum at day 42. Diet supplemented with E. faecium PNC01 increased the relative abundance of Firmicutes and Lactobacillus, decreased the relative abundance of Bacteroides in the cecal microbiota. Conclusion E. faecium PNC01 replaced antibiotics to reduce the feed conversion rate. Furthermore, E. faecium PNC01 improved intestinal morphology and altered the composition of microbiota in the cecum to reduce feed conversion rate. Thus, it can be used as an alternative for antibiotics in broiler production to avoid the adverse impact of antibiotics by altering the gut microbiota. Graphic Abstract

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Micro-encapsulated sodium butyrate attenuates oxidative stress induced by corticosterone exposure and modulates apoptosis in intestinal mucosa of broiler chickens

Animal Production Science ◽

10.1071/an13348 ◽

2015 ◽

Vol 55 (5) ◽

pp. 587 ◽

Cited By ~ 5

Author(s):

Y. Jiang ◽

W. H. Zhang ◽

F. Gao ◽

G. H. Zhou

Keyword(s):

Oxidative Stress ◽

Epithelial Cells ◽

Intestinal Mucosa ◽

Sodium Butyrate ◽

Broiler Chickens ◽

Intestinal Epithelial Cells ◽

Control Diet ◽

Jejunal Mucosa ◽

Intestinal Epithelial ◽

Apoptosis Index

The aim of the present study was to investigate the effects of micro-encapsulated sodium butyrate (MSB) on oxidative stress and apoptosis induced by dietary corticosterone (CORT) in the intestinal mucosa of broiler chickens. In total, 120 1-day-old male broilers (Arbor Acres) were randomly allocated to two treatment groups and were fed on a control diet (without MSB) or 0.4 g MSB/kg diet. Each treatment had six replicates with five chickens each. From 7 days of age onward, 50% of the chickens in each dietary treatment were subjected to CORT treatment (30 mg/kg of diet). The experimental period was 21 days. The results showed that CORT administration decreased (P < 0.001) feed intake and bodyweight gain and increased (P < 0.001) feed to gain ratio (F : G) of broiler chickens. The dietary MSB supplementation decreased (P < 0.01) F : G and there was an interaction between MSB and CORT on F : G (P < 0.05). Moreover, the activities of superoxide dismutase, glutathione peroxidase and catalase in intestinal mucosa were decreased (P < 0.01 or P < 0.001), and the concentrations of malondialdehyde in the intestinal mucosa were elevated (P < 0.01) by CORT administration. In contrast, treatment of MSB increased (P < 0.01) the catalase activities in duodenal and jejunal mucosa and decreased (P < 0.01) the malondialdehyde concentrations in duodenal mucosa. Higher apoptosis index and lower mRNA expressions of bcl-2 in intestinal epithelial cells were induced (P < 0.05) by CORT treatment. However, MSB decreased (P < 0.05) the apoptosis index and increased the bcl-2 expression. These results suggest that dietary MSB can partially attenuate oxidative stress induced by CORT treatment and inhibit apoptosis of intestinal epithelial cells in broiler chickens.

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Immunocytochemical localization of ornithine transcarbamylase in rat intestinal mucosa. Light and electron microscopic study.

Journal of Histochemistry & Cytochemistry ◽

10.1177/36.1.3275711 ◽

1988 ◽

Vol 36 (1) ◽

pp. 29-35 ◽

Cited By ~ 11

Author(s):

Y Hamano ◽

H Kodama ◽

M Yanagisawa ◽

Y Haraguchi ◽

M Mori ◽

...

Keyword(s):

Electron Microscopy ◽

Small Intestine ◽

Epithelial Cells ◽

Light Microscopy ◽

Intestinal Mucosa ◽

Intestinal Epithelial Cells ◽

Protein A ◽

Intestinal Epithelial ◽

Electron Microscopic ◽

Small Intestinal

We investigated light and electron microscopic localization of ornithine transcarbamylase (OTC) in rat intestinal mucosa. In the immunoblotting assay of OTC-related protein, a single protein band with a molecular weight of about 36,500 is observed in extracts of liver and small intestinal mucosa but is not observed in those of stomach and large intestine. For light microscopy, tissue slices of the digestive system were embedded in Epon and stained by using anti-bovine OTC rabbit IgG and the immunoenzyme technique. For electron microscopy, slices of these and the liver tissues were embedded in Lowicryl K4M and stained by the protein A-gold technique. By light microscopy, the absorptive epithelial cells of duodenum, jejunum, and ileum stained positively for OTC, but stomach, large intestine, rectum, and propria mucosa of small intestine were not stained. Electron microscopy showed that gold particles representing the antigenic sites for OTC were confined to the mitochondrial matrix of hepatocytes and small intestinal epithelial cells. However, the enzyme was detected in mitochondria of neither liver endothelial cells, submucosal cells of small intestine, nor large intestinal epithelial cells. Labeling density of mitochondria in the absorptive epithelial cells of duodenum, jejunum, and ileum was about half of that in liver cells.

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Interaction of Bifidobacterium bifidum LMG13195 with HT29 Cells Influences Regulatory-T-Cell-Associated Chemokine Receptor Expression

Applied and Environmental Microbiology ◽

10.1128/aem.07581-11 ◽

2012 ◽

Vol 78 (8) ◽

pp. 2850-2857 ◽

Cited By ~ 41

Author(s):

Patricia López ◽

Irene González-Rodríguez ◽

Borja Sánchez ◽

Patricia Ruas-Madiedo ◽

Ana Suárez ◽

...

Keyword(s):

T Cell ◽

Epithelial Cells ◽

Intestinal Mucosa ◽

Chemokine Receptor ◽

Intestinal Epithelial Cells ◽

Bifidobacterium Bifidum ◽

Receptor Expression ◽

Intestinal Epithelial ◽

Content Type ◽

Ht29 Cells

ABSTRACTProbiotics play an important role in the maintenance of the gastrointestinal barrier. In addition to direct effects on mucosal integrity, the interaction with the intestinal mucosa may have an active immunoregulatory effect. In the present work, we exposed HT29 intestinal epithelial cells to twoBifidobacteriumspecies to determine their effect on gene expression profile, enterocyte monolayer integrity, and T-cell response.Bifidobacterium breveIPLA 20004 triggered a more pronounced increase in the transepithelial resistance of the enterocyte monolayer thanBifidobacterium bifidumLMG13195. The transcriptome profile of HT29 cells cultured in the presence ofB. bifidumLMG13195 showed an increased expression of immune mediators and, interestingly, chemotactic molecules (CXCL10, CCL20, CXCL11 and CCL22) able to recruit lymphocytes. Since regulatory T cells (Treg cells) may express receptors for specific chemokines, we cultured peripheral blood mononuclear cells with supernatants of HT29 cells previously treated withBifidobacteriumstrains and analyzed FOXP3 and CD25 Treg markers and CCR6, CXCR3, CCR4, and CCR3 expression on CD4+lymphocytes. The proportion of CD25highFOXP3+cells was significantly increased after culture withB. bifidumLMG13195-conditioned HT29 supernatant. Moreover, this treatment led to the largest amount of CCR6+CXCR3−CCR4+CCR3+CD4+cells expressing high levels of CD25, corresponding to the Treg population. These results suggest that soluble factors secreted afterB. bifidumLMG13195 contact with intestinal epithelial cells favored the generation of CD4+CD25highlymphocytes expressing chemokine receptor Treg markers, thus making possible their recruitment to the intestinal mucosa.

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Entry of spores into intestinal epithelial cells contributes to recurrence of Clostridioides difficile infection

Nature Communications ◽

10.1038/s41467-021-21355-5 ◽

2021 ◽

Vol 12 (1) ◽

Cited By ~ 1

Author(s):

Pablo Castro-Córdova ◽

Paola Mora-Uribe ◽

Rodrigo Reyes-Ramírez ◽

Glenda Cofré-Araneda ◽

Josué Orozco-Aguilar ◽

...

Keyword(s):

Epithelial Cells ◽

Mouse Model ◽

Intestinal Mucosa ◽

Intestinal Epithelial Cells ◽

Intestinal Barrier ◽

Intestinal Epithelial ◽

Spore Surface ◽

Clostridioides Difficile ◽

Clostridioides Difficile Infection ◽

Infection Recurrence

AbstractClostridioides difficile spores produced during infection are important for the recurrence of the disease. Here, we show that C. difficile spores gain entry into the intestinal mucosa via pathways dependent on host fibronectin-α5β1 and vitronectin-αvβ1. The exosporium protein BclA3, on the spore surface, is required for both entry pathways. Deletion of the bclA3 gene in C. difficile, or pharmacological inhibition of endocytosis using nystatin, leads to reduced entry into the intestinal mucosa and reduced recurrence of the disease in a mouse model. Our findings indicate that C. difficile spore entry into the intestinal barrier can contribute to spore persistence and infection recurrence, and suggest potential avenues for new therapies.

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Analysis of Intestinal Mucosa Integrity and GLP-2 Gene Functions upon Porcine Epidemic Diarrhea Virus Infection in Pigs

Animals ◽

10.3390/ani11030644 ◽

2021 ◽

Vol 11 (3) ◽

pp. 644

Author(s):

Yajing Zhou ◽

Zhanshi Ren ◽

Shuai Zhang ◽

Haifei Wang ◽

Shenglong Wu ◽

...

Keyword(s):

Epithelial Cells ◽

Intestinal Mucosa ◽

Porcine Epidemic Diarrhea Virus ◽

Intestinal Epithelial Cells ◽

Mrna Level ◽

Mucosal Barrier ◽

Intestinal Epithelial ◽

Intestinal Mucosal Barrier ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea

Porcine epidemic diarrhea virus (PEDV) infects intestinal epithelial cells, destroys the intestinal mucosal barrier and then causes diarrhea in piglets. Glucagon-like peptide-2 (GLP-2) is a specific intestinal growth hormone that promotes the repair of damaged intestinal mucosa and improves the intestinal barrier. In this study, we investigated the functions of porcine GLP-2 gene in regulating PEDV infection. The intestinal tissues with damaged intestinal structures caused by PEDV infection were first confirmed and collected. Expression analysis indicated that the GLP-2 gene was expressed in the duodenum, jejunum and ileum tissues, and the mRNA level was significantly down-regulated in jejunum and ileum of piglets with damaged intestinal mucosa. Infection of PEDV to porcine small intestinal epithelial cells in vitro showed that GLP-2 gene was significantly decreased, which was consistent with the expression pattern in intestinal tissues. In addition, we silenced the GLP-2 gene by shRNA interfering and found that the copy numbers of PEDV were remarkably increased in the GLP-2 gene silencing cells. Our findings suggest that the GLP-2 gene was potentially involved in regulating PEDV infection and in maintaining the integrity of the intestinal mucosal barrier structure, which could contribute to our understanding of the mechanisms of PEDV pathogenesis and provide a theoretical basis for the identification and application of resistant genes in pig selective breeding for porcine epidemic diarrhea.

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P010 GSDMB-mediated pyroptosis exacerbates intestinal inflammation by destroying intestinal epithelium in Crohn’s disease

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jjab076.139 ◽

2021 ◽

Vol 15 (Supplement_1) ◽

pp. S129-S131

Author(s):

W Gong ◽

P Liu ◽

J Ren

Keyword(s):

T Cells ◽

Crohn’S Disease ◽

Epithelial Cells ◽

Intestinal Mucosa ◽

Cd8 T Cells ◽

Intestinal Inflammation ◽

Disease Risk ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Epithelial Injury

Abstract Background Intestinal epithelial injury acts an essential role in the pathogenesis and development of Crohn’s disease (CD). Recent studies indicated that gasdermin D (GSDMD) mediated pyroptosis in intestinal epithelial cell (IEC) contributes to the epithelial injury during intestinal inflammation. However, how gasdermin B (GSDMB) mediated pyroptosis regulates intestinal epithelial injury in CD remains unknown. Methods We studied the characteristics of GSDMB mediated pyroptosis in CD patients and intestinal epithelial cells (Caco-2, HT-29 and primary IECs). The CD8+ T cells were extracted from intestinal mucosa of CD patients and health controls, and then co cultured with normal primary IECs to observe the pyroptosis of IECs. We further analyzed the CD8+ T cell subsets that promote pyroptosis in intestinal biopsies of CD patients. In addition, we screened out four single nucleotide polymorphism (SNP) of GSDMB that related to disease risk of CD in the public IBD exomes database (https://ibd.broadinstitute.org/), and investigated their effects on pyroptosis. Results GSDMB mediated pyroptosis was notably increased in intestinal mucosa of active human CD, and only existed in intestinal epithelial cells (Figure 1). Granzyme A (GrzA) initiated the killing results by the cleavage of GSDMB in intestinal epithelial cells, and interferon gamma (IFN-γ) up-regulated GSDMB expression in intestinal epithelial cells and promoted pyroptosis (Figure 2). GSDMB in primary IECs was significantly cleaved when co cultured with CD8+ T cells from active CD patients, while the phenomenon weakened a lot when GrzA in CD8+ T cells or GSDMB in epithelial cells was knocked down (Figure 3). We also found that IL26 possive CD8+ T cells were the main CD8+ T cells subsets secreting GrzA (Figure 3). We screened out two SNP of GSDMB related to increased disease risk of CD (rs2305480 and rs11078928) and two related to decreased disease risk (rs35104165 and rs143933205), and constructed these four mutant plasmids. Compared with wild type of GSDMB, rs2305480 promoted the cleavage while rs35104165 blocked the cleavage (Figure 4). Conclusion GSDMB-mediated pyroptosis results in intestinal epithelial injury, which will exacerbate intestinal inflammation. Modulation of the GSDMB-mediated pyroptosis emerges as a potential therapeutic strategy to target epithelium damage and treat CD.

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M1780 Human Intestinal Epithelial Cells: Actors of the Proteolytic Balance of Intestinal Mucosa

Gastroenterology ◽

10.1016/s0016-5085(10)61923-3 ◽

2010 ◽

Vol 138 (5) ◽

pp. S-417-S-418

Author(s):

Jean-Paul Motta ◽

Laurent Magne ◽

Jean-Michel Salllenave ◽

Michel Chignard ◽

Nathalie Vergnolle

Keyword(s):

Epithelial Cells ◽

Intestinal Mucosa ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Human Intestinal Epithelial Cells

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Glycerolipid Biosynthesis in Isolated Rat Intestinal Epithelial Cells

Canadian Journal of Biochemistry ◽

10.1139/o75-138 ◽

1975 ◽

Vol 53 (9) ◽

pp. 1010-1019 ◽

Cited By ~ 16

Author(s):

P. J. A. O'Doherty ◽

A. Kuksis

Keyword(s):

Fatty Acids ◽

Epithelial Cells ◽

Free Fatty Acids ◽

Intestinal Mucosa ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Trypan Blue Exclusion ◽

Low Degree ◽

Total Capacity ◽

Everted Sacs

Intestinal epithelial cells were prepared from fasted rats by dispersion with collagenase (EC 3.4.24.3). The structural and metabolic integrity of the cells was verified by electron microscopy, a high percentage of Trypan Blue exclusion, a low degree of release of lactate dehydrogenase (EC 1.1.1.27) in the medium, and by the retention of sensitivity to agents known to modify metabolic and transport activity in everted sacs of intestinal mucosa. The isolated intestinal epithelial cells were used to study glycerolipid biosynthesis from glucose, glycerol, 2-monoacylglycerol, and free fatty acids. The cells actively incorporated the labeled precursors into glycerolipids without specific cofactor requirements. Addition of fatty acids stimulated the incorporation of both glucose and glycerol into triacylglycerols and glycerophospholipids, the greatest effect being observed with palmitate. The stimulation of monoacylglycerol acylation appeared to depend on both the nature of the monoacylglycerol and fatty acid supplied. Stereospecific analyses of the diacylglycerols formed from 2-monoacylglycerols and free fatty acids showed that 1,2-diacyl-sn-glycerols (62–70%) were the major and that 2,3-diacyl-sn-glycerols (30–38%) the minor intermediates in triacylglycerol biosynthesis. The data indicate that isolated intestinal epithelial cells exhibit a total capacity of glycerolipid synthesis and a stereochemical course of reaction which is comparable to that observed for triacylglycerol formation in everted sacs of intestinal mucosa, but much less specific than that seen in microsomal preparations of intestinal mucosa.

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Primary culture of salamander intestinal epithelial cells from nests: whole cell Na+ currents induced by valine

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1994.267.1.g59 ◽

1994 ◽

Vol 267 (1) ◽

pp. G59-G66

Author(s):

J. F. White

Keyword(s):

Epithelial Cells ◽

Intestinal Mucosa ◽

Intestinal Epithelial Cells ◽

Intestinal Cells ◽

Small Intestinal Mucosa ◽

Intestinal Epithelial ◽

Patch Clamp Technique ◽

Whole Cell ◽

Inward Current ◽

Small Intestinal

Methods are described for isolating the cell nests, subepithelial clusters of germinative cells, from salamander intestinal mucosa and for growing the nests in culture into polarized monolayers of intestinal epithelial cells. Cells were viable in culture for up to 3 wk. The capacity of the monolayer cells to engage in membrane transport was evaluated using the patch-clamp technique in the whole cell mode. L-Valine (25 mM) induced an inward current in small intestinal cells of 25.8 +/- 5.7 pA and depolarized the cell membrane 14.5 +/- 1.6 mV. L-Alanine and L-phenylalanine were similarly effective, whereas D-valine was ineffective. The Km of the transporter for valine was 90 mM. Replacement of bath Na with tris(hydroxymethyl)aminomethane eliminated the inward current induced by valine. The basal (solute-independent) inward current was also reduced by Na+ replacement. Glucose did not induce a Na+ current. In contrast to the effect of valine on small intestinal cells, large intestinal cells were unresponsive to valine. It is concluded that the cultured small intestinal cells possess Na-amino acid but not Na-sugar cotransport. This profile of behavior is characteristic of undifferentiated small intestinal cells. Primary cultures of salamander small intestinal cells should be useful for studying enterocyte function and the developmental biology of the small intestinal mucosa.

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