Gene expression in the chicken caecum is dependent on microbiota composition

Gut microbiota is of considerable importance for each host. Despite this, germ-free animals can be obtained and raised to sexual maturity and consequences of the presence or absence of gut microbiota on gene expression of the host remain uncharacterised. In this study, we performed an unbiased study of protein expression in the caecum of germ-free and colonised chickens. The major difference between these two groups was in the expression of immunoglobulins which were essentially absent in the germ-free chickens. Microbiota also caused a minor decrease in the expression of focal adhesion and extracellular matrix proteins and an increase in the expression of argininosuccinate synthase ASS1, redox potential sensing, fermentative metabolic processes and detoxification systems represented by sulfotransferases SULT1C3 or SULT1E1. Since we also analysed expression in the caecum of E. coli Nissle and E. faecium DSM7134 mono-associated chickens, we concluded that at least immunoglobulin expression and expression of cystathionine synthase (CBS) was dependent on microbiota composition with E. coli Nissle stimulating more immunoglobulin and PIGR expression and E. faecium DSM7134 stimulating more CBS expression. Gut microbiota and its composition therefore affected protein expression in the chicken caecum though except for immunoglobulin production, the remaining differences were unexpectedly low

Entrepreneurial orientation and the business performance of SMEs: a quantitative study from the Netherlands

Additional file 5. Macrophage proteins responding to in vivo infection with S. Enteritidis

The response of porcine monocyte derived macrophages and dendritic cells to SalmonellaTyphimurium and lipopolysaccharide

BACKGROUND: Following infection and initial multiplication in the gut lumen, Salmonella Typhimurium crosses the intestinal epithelial barrier and comes into contact with cells of the host immune system. Mononuclear phagocytes which comprise macrophages and dendritic cells (DC) are of key importance for the outcome of Salmonella infection. Although macrophages and DC may differentiate from a common precursor, their capacities to process and present antigen differ significantly. In this study, we therefore compared the response of porcine macrophages and DC differentiated from peripheral blood monocytes to S. Typhimurium and one of the most potent bacterial pathogen associated molecular patterns, bacterial lipopolysaccharide. To avoid any bias, the expression was determined by protein LC-MS/MS and verified at the level of transcription by quantitative RT-PCR. RESULTS: Within 4 days of culture, peripheral blood monocytes differentiated into two populations with distinct morphology and expression of MHC II. Mass spectrometry identified 446 proteins in macrophages and 672 in DC. Out of these, 433 proteins were inducible in macrophages either after infection with S. Typhimurium or LPS exposure and 144 proteins were inducible in DC. The expression of the 46 most inducible proteins was verified at the level of transcription and the differential expression was confirmed in 22 of them. Out of these, 16 genes were induced in both cell types, 3 genes (VCAM1, HMOX1 and Serglycin) were significantly induced in macrophages only and OLDLR1 and CDC42 were induced exclusively in DC. Thirteen out of 22 up-regulated genes contained the NF-kappaB binding site in their promoters and could be considered as either part of the NF-kappaB feedback loop (IkappaBalpha and ISG15) or as NF-kappaB targets (IL1beta, IL1alpha, AMCF2, IL8, SOD2, CD14, CD48, OPN, OLDLR1, HMOX1 and VCAM1). CONCLUSIONS: The difference in the response of monocyte derived macrophages and DC was quantitative rather than qualitative. Despite the similarity of the responses, compared to DC, the macrophages responded in a more pro-inflammatory fashion. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12917-014-0244-1) contains supplementary material, which is available to authorized users

Transient and Prolonged Response of Chicken Cecum Mucosa to Colonization with Different Gut Microbiota

In this study we determined protein and gene expression in the caeca of newly hatched chickens inoculated with cecal contents sourced from hens of different ages. Over 250 proteins exhibited modified expression levels in response to microbiota inoculation. The most significant inductions were observed for ISG12-2, OASL, ES1, LYG2, DMBT1-L, CDD, ANGPTL6, B2M, CUZD1, IgM and Ig lambda chain. Of these, ISG12-2, ES1 and both immunoglobulins were expressed at lower levels in germ-free chickens compared to conventional chickens. In contrast, CELA2A, BRT-2, ALDH1A1, ADH1C, AKR1B1L, HEXB, ALDH2, ALDOB, CALB1 and TTR were expressed at lower levels following inoculation of microbiota. When chicks were given microbiota preparations from different age donors, the recipients mounted differential responses to the inoculation which also differed from the response profile in naturally colonised birds. For example, B2M, CUZD1 and CELA2A responded differently to the inoculation with microbiota of 4- or 40-week-old hens. The increased or decreased gene expression could be recorded 6 weeks after the inoculation of newly hatched chickens. To characterise the proteins that may directly interact with the microbiota we characterised chicken proteins that co-purified with the microbiota and identified a range of host proteins including CDD, ANGPTL6, DMBT1-L, MEP1A and Ig lambda. We propose that induction of ISG12-2 results in reduced apoptosis of host cells exposed to the colonizing commensal microbiota and that CDD, ANGPTL6, DMBT1-L, MEP1A and Ig lambda reduce contact of luminal microbiota with the gut epithelium thereby reducing the inflammatory response

The Early Innate Response of Chickens to <i>Salmonella enterica</i> Is Dependent on the Presence of O-Antigen but Not on Serovar Classification

<div><p><i>Salmonella</i> vaccines used in poultry in the EU are based on attenuated strains of either <i>Salmonella</i> serovar Enteritidis or Typhimurium which results in a decrease in <i>S</i>. Enteritidis and <i>S</i>. Typhimurium but may allow other <i>Salmonella</i> serovars to fill an empty ecological niche. In this study we were therefore interested in the early interactions of chicken immune system with <i>S</i>. Infantis compared to <i>S</i>. Enteritidis and <i>S</i>. Typhimurium, and a role of O-antigen in these interactions. To reach this aim, we orally infected newly hatched chickens with 7 wild type strains of <i>Salmonella</i> serovars Enteritidis, Typhimurium and Infantis as well as with their <i>rfaL</i> mutants and characterized the early <i>Salmonella</i>-chicken interactions. Inflammation was characterized in the cecum 4 days post-infection by measuring expression of 43 different genes. All wild type strains stimulated a greater inflammatory response than any of the <i>rfaL</i> mutants. However, there were large differences in chicken responses to different wild type strains not reflecting their serovar classification. The initial interaction between newly-hatched chickens and <i>Salmonella</i> was found to be dependent on the presence of O-antigen but not on its structure, i.e. not on serovar classification. In addition, we observed that the expression of calbindin or aquaporin 8 in the cecum did not change if inflammatory gene expression remained within a 10 fold fluctuation, indicating the buffering capacity of the cecum, preserving normal gut functions even in the presence of minor inflammatory stimuli.</p></div

Transient and prolonged response of chicken caecum mucosa to the colonization with different gut microbiota

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PCA plot of the chickens clustered according to their gene expression in the cecum and heat map correlation coefficients between factor 1 and individual gene expression.

<p>Open black circles, <i>S</i>. Enteritidis 147; Open blue circles, <i>S.</i> Enteritidis G1481; open black squares, <i>S</i>. Typhimurium LT2; open blue squares, <i>S</i>. Typhimurium 2002; open red squares, <i>S</i>. Typhimurium 2454; open black triangles, <i>S</i>. Infantis 1516; open blue triangles, <i>S</i>. Infantis 514; closed black circles, <i>S</i>. Enteritidis 147 <i>rfaL</i> mutant; closed black squares, <i>S</i>. Typhimurium LT2 <i>rfaL</i> mutant; closed black triangles, <i>S</i>. Infantis 1516 <i>rfaL</i> (I) mutant; closed blue triangles, <i>S</i>. Infantis 1516 <i>rfaL</i> (II) mutant; closed red triangles, <i>S</i>. Infantis 1516 <i>rfaL</i> (III) mutant. symbol “plus”, non-infected chickens. PCA also showed that a single factor explained nearly 80% of the variation in the chicken response. This factor was the scope of inflammation itself as high and significant correlations were observed between the expression of individual genes and the positioning of corresponding chickens along X axis. Genes are arranged from the most positively correlated to the most negatively correlated ones.</p

Cytokine gene expression in the cecum of orally infected chickens.

<p>Columns represent geometric means of the relative expressions of respective genes. Vertical bars represent 95% confidence intervals regarding the geometric means. Superscripts above columns denote statistically significant differences among groups (columns sharing the same superscript are not significantly different from each other, columns that have no superscript in common are significantly different from each other). NI, expression in the non-infected chickens. SE, expression in the chickens infected with <i>S</i>. Enteritidis 147. STM, expression in the chickens infected with <i>S</i>. Typhimurium LT2. SI, expression in the chickens infected with <i>S</i>. Infantis 1516. SE <i>rfaL</i>, expression in the chickens infected with <i>S</i>. Enteritidis <i>rfaL</i> mutant. STM <i>rfaL</i>, expression in the chickens infected with <i>S</i>. Typhimurium <i>rfaL</i> mutant. SI <i>rfaL</i>, expression in the chickens infected with <i>S</i>. Infantis <i>rfaL</i> (I) mutant. Mind logarithmic scaling of Y-axis.</p