BioProject

The host response to S.typhimurium infections in ileum was studied after infection of piglets. More...

The host response to S.typhimurium infections in ileum was studied after infection of piglets. mRNA and miRNA expression studies were performed to identify interactions of miRNAs and regulated mRNAs in context of mammalian intestinal Salmonella infection using a piglet model. Overall design: Intestinal samples (~ 2 cm circle segments) were taken at time points 3h, 3d and 28d post infection from ileum of ten S.typhimurium infected piglets (S), ten S.typhimurium infected piglets and additional treated with probiotic strain E.faecium NCIMB 10415 (SP) and five healthy control piglets(C) (EUROC x Pietrain). The piglets were weaned at the age of 28 days and infected with S. typhimurium. Samples were quick-frozen in liquid nitrogen and stored at -80 °C. In order to obtain representative measurements in each intestinal locus, three cross sections of approximately 2 mm out of the 2 cm segment of frozen intestine were examined. These 3 sections were pooled and total RNA was isolated from samples using an automated homogenizer (FastPrep Instrument, MP Biomedicals, Heidelberg, Germany) and the mirVana miRNA Isolation Kit (Applied Biosystems, Darmstadt, Germany), according to the manufacturer’s protocol. The RNA quality and quantity of all samples were proven using the Agilent 2100 Bioanalyzer and the RNA 6000 Nano Kits (Agilent, Waldbronn, Germany) and the Nanodrop 1000 Spectrophotometer (Thermo Scientific, MA, USA). For microarray analysis pooled samples of each experimental group were prepared ana partes aequalis from individually isolated total RNA samples, which were collected at the same time point in each group. Common reference was designed from equal amounts of total RNA of each pool and additional total RNA from ascending colon. Less...