Project description:The response of chicken to non-typhoidal Salmonella infection is becoming well characterised but the role of particular cell types in this response is still far from being understood. Therefore, in this study we characterised the response of chicken embryo fibroblasts (CEFs) to infection with two different S. Enteritidis strains by microarray analysis. The expression of chicken genes identified as significantly up- or down-regulated (≥3-fold) by microarray analysis was verified by real-time PCR followed by functional classification of the genes and prediction of interactions between the proteins using Gene Ontology and STRING Database. Finally the expression of the newly identified genes was tested in HD11 macrophages and in vivo in chickens. Altogether 19 genes were induced in CEFs after S. Enteritidis infection. Twelve of them were also induced in HD11 macrophages and thirteen in the caecum of orally infected chickens. The majority of these genes were assigned different functions in the immune response, however five of them (LOC101750351, K123, BU460569, MOBKL2C and G0S2) have not been associated with the response of chicken to Salmonella infection so far. K123 and G0S2 were the only 'non-immune' genes inducible by S. Enteritidis in fibroblasts, HD11 macrophages and in the caecum after oral infection. The function of K123 is unknown but G0S2 is involved in lipid metabolism and in β-oxidation of fatty acids in mitochondria. Increased levels of G0S2 might decrease the availability of fatty acids to mitochondria. In non-professional phagocytes such as CEFs, this may lead to the dysfunction of mitochondria, apoptosis of CEFs and release of intracellular Salmonella. In professional phagocytes, G0S2 might be involved in the control of mitochondrial respiration, resulting in a decrease of reactive oxygen species as respiration by-products and lower damage to tissue. In this study we were interested whether chicken embryo fibroblast (CEFs) respond to S. Enteritidis infection and to what extent their response differs from that of other cells and caecal tissue. To address this, we characterised the gene expression of CEFs after infection with two different wild-type S. Enteritidis strains of poultry origin - SE 147 and SE 11 - using Agilen custom 8×15K microarrays. In total, 13,681 probes were designed to characterise the expression of ~9,000 transcripts of Gallus gallus.
Project description:The response of chicken to non-typhoidal Salmonella infection is becoming well characterised but the role of particular cell types in this response is still far from being understood. Therefore, in this study we characterised the response of chicken embryo fibroblasts (CEFs) to infection with two different S. Enteritidis strains by microarray analysis. The expression of chicken genes identified as significantly up- or down-regulated (≥3-fold) by microarray analysis was verified by real-time PCR followed by functional classification of the genes and prediction of interactions between the proteins using Gene Ontology and STRING Database. Finally the expression of the newly identified genes was tested in HD11 macrophages and in vivo in chickens. Altogether 19 genes were induced in CEFs after S. Enteritidis infection. Twelve of them were also induced in HD11 macrophages and thirteen in the caecum of orally infected chickens. The majority of these genes were assigned different functions in the immune response, however five of them (LOC101750351, K123, BU460569, MOBKL2C and G0S2) have not been associated with the response of chicken to Salmonella infection so far. K123 and G0S2 were the only 'non-immune' genes inducible by S. Enteritidis in fibroblasts, HD11 macrophages and in the caecum after oral infection. The function of K123 is unknown but G0S2 is involved in lipid metabolism and in β-oxidation of fatty acids in mitochondria. Increased levels of G0S2 might decrease the availability of fatty acids to mitochondria. In non-professional phagocytes such as CEFs, this may lead to the dysfunction of mitochondria, apoptosis of CEFs and release of intracellular Salmonella. In professional phagocytes, G0S2 might be involved in the control of mitochondrial respiration, resulting in a decrease of reactive oxygen species as respiration by-products and lower damage to tissue.
Project description:1-day old chickens were orally inoculated with 1e5 c.f.u. Salmonella enteritidis. 24 hours after infection RNA from the jejunum was analysed. Two different microarray platforms were used, a homemade cDNA microarray and the Affymetrix GeneChip Chicken Genome Array Keywords: host response; genetic background
Project description:Poultry products are an important source of Salmonella enterica. An effective way to reduce food poisoning due to Salmonella would be to breed chickens more resistant to Salmonella. Unfortunately resistance to Salmonella is a complex trait with many factors involved. To learn more about Salmonella resistance mechanisms in young chickens, a cDNA microarray analysis was performed to compare gene expression profiles between a Salmonella susceptible and a more resistant chicken line. Newly hatched chickens were orally infected with S. enterica serovar Enteritidis. Since the intestine is the first barrier the bacteria encountersbacteria encounter after oral inoculation, gene expression was investigated in the intestine, from day 1 until day 21 post infection. Differences in gene expression between the susceptible and resistant chicken line were found in control as well as Salmonella infected conditions. In response to the Salmonella infection, the expression of different sets of genes seemed to be affected in the jejunum of the two chicken lines. In the susceptible line this included genes that affect T-cell activation, whereas in the more resistant line, at day 1, macrophage activation seemed to be more affected. At day 7 and 9 most gene expression differences between the two chicken lines were identified under control conditions, indicating a difference in the intestinal development between the two chicken lines which might be linked to the difference in Salmonella susceptibility. The findings in this study have lead to the identification of novel genes and possible cellular pathways of the host involved in Salmonella susceptibility. Keywords: timecourse, disease
Project description:To provide insight into the systemic metabolic effects of S. enteritidis infection, liver samples were harvested 10 days post infection from broiler hens. Hepatic global gene expression levels were assessed using a chicken 44K Agilent microarray. Forty-four genes were differentially expressed at a significance level of q-value < 0.05. One hundred eighty-three genes were differentially expressed at a suggestive significance level of q-value < 0.1. A predominance of down-regulation existed among significantly differentially expressed genes. A cell morphology, cell cycle, organismal injury and abnormalities network and a metabolic disease, cardiovascular system development and function, and urological disease network were created from the differentially expressed genes. Apoptosis, electron transport, peptidase activity, vein constriction, cell differentiation, IL-2 signaling, Jak-Stat signaling, B-cell receptor signaling, GDP/GTP exchange, and cytokine suppression were among the functions of the differentially expressed genes that were down-regulated in response to S. enteritidis. The effects of S. enteritidis infection on the liver transcriptome profiles of broilers reflect a predominance of down-regulation of genes involved with metabolic, cell morphology, cell cycle, urological disease, and organismal injury functions. These results provide insight into important systemic metabolic mechanisms that are active in the chicken liver in response to S. enteritidis infection.
Project description:Salmonella enteritidis is suggested to translocate in the small intestine. Previously we identified that prebiotics, fermented in the colon, increased Salmonella translocation in rats, suggesting involvement of the colon in translocation. Effects of Salmonella on colonic gene expression in vivo are largely unknown. The aim of this study was to characterize time dependent Salmonella induced changes of colonic mucosal gene expression in rats using whole genome microarrays. Rats were orally infected with Salmonella enteritidis to mimic a foodbore infection and colonic gene expression was determined at day 1, 3 and 6 post-infection (n=8 per timepoint). Agilent rat whole genome microarray (G4131A Agilent Technologies) were used. Results indicate that colon is clearly a target tissue for Salmonella considering the abundant changes in mucosal gene expression observed. Keywords: Time point infection study, colon mucosa, Rat
Project description:Global gene expression profiling in spleen in chicken inoculated with S. enteritidis and control was conducted to screen differentially expressed genes and identify the key gene, signalling pathways and important biological processes that related to the infection of S. enteritidis
Project description:To provide insight into the systemic metabolic effects of S. enteritidis infection, liver samples were harvested 10 days post infection from broiler hens. Hepatic global gene expression levels were assessed using a chicken 44K Agilent microarray. Forty-four genes were differentially expressed at a significance level of q-value < 0.05. One hundred eighty-three genes were differentially expressed at a suggestive significance level of q-value < 0.1. A predominance of down-regulation existed among significantly differentially expressed genes. A cell morphology, cell cycle, organismal injury and abnormalities network and a metabolic disease, cardiovascular system development and function, and urological disease network were created from the differentially expressed genes. Apoptosis, electron transport, peptidase activity, vein constriction, cell differentiation, IL-2 signaling, Jak-Stat signaling, B-cell receptor signaling, GDP/GTP exchange, and cytokine suppression were among the functions of the differentially expressed genes that were down-regulated in response to S. enteritidis. The effects of S. enteritidis infection on the liver transcriptome profiles of broilers reflect a predominance of down-regulation of genes involved with metabolic, cell morphology, cell cycle, urological disease, and organismal injury functions. These results provide insight into important systemic metabolic mechanisms that are active in the chicken liver in response to S. enteritidis infection. Sixteen broiler hens of approximately 5 months (8 challenged orally with 1x108 S. enteritidis on each of three sequential days and 8 mock-challenged with phosphate buffered saline) were used. Livers were harvested at 10 days post infection and stored in RNALater (Ambion).