Evaluation of the potential of non-starch soluble polysaccharides from 3 algae to modulate the immune response and metabolic state in the intestine
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ABSTRACT: Non-starch soluble polysaccharides (NSPs) produced by yeasts are used in animal nutrition to improve health and performance. However, the magnitude of the effect may be dependent upon the quantity and the composition of the polysaccharides. As seaweeds are attractive sources of NSPs, this study was set up to evaluate their potential to improve intestinal health. The effect of NSP extracts prepared from Saccharomyces cerevisiae containing β-glucan and mannan (PSY1, positive control) or a mixture of mannanoligosaccharides (PSY2, positive control), micro algae containing β-glucan (PSA1), brown macro algae containing fucoidan and laminarin (PSA2), and green macro algae containing ulvan (PSA3) on intestinal porcine epithelial cells J2 (IPEC-J2) was studied in the presence and absence of the enterotoxigenic bacterium Escherichia coli k99 strain (ETEC) as an in vitro challenge. The E.coli-k99 strain with adhesion factor F41 (41/32) was isolated from a mastitis-infected udder. In addition, a mixed extract prepared from vegatal orgin supplemented with phenolic compounds from vegetal origin, zinc and selenium (9631), and ZnO were tested to compare responses to NSP extracts. Gene expression was measured in IPEC-J2 cells after 2 and 6 hours of incubation using “whole genome” porcine microarrays (submission as a conference paper at the SEAGRICULTURE 2017 6th International Seaweed Conference).
Project description:Deoxynivalenol (DON) frequently detected in a wide range of foods and feeds, inducing cytotoxicity to animals and humans. N6-methyladenosine (m6A) is an important epitranscriptomic marker with high abundance in eukaryotic mammals mRNAs. However, the role of the m6A methylomes in DON damage is still poorly understood. Here, we investigated the m6A transcriptome-wide profile in intestinal porcine epithelial cells (IPEC-J2) with and without 1000 ng/mL DON treatment via m6A sequencing and RNA sequencing. In total, 5406 new m6A peaks appeared with the disappearance of 2615 peaks in DON-induced IPEC-J2. The unique m6A-modified genes in DON-induced IPEC-J2 were associated with TNF signaling pathway. We identified 733 differentially expressed mRNA transcripts with hyper-methylated or hypo-methylated m6A peaks between DON-induced IPEC-J2 and normal IPEC-J2. Protein interaction network analysis and qPCR validation suggested that CSF2 probably acts as a promising new target for combating DON damage in IPEC-J2. Our first report of m6A transcriptome-wide map of IPEC-J2 cells presented here provides a starting roadmap for uncovering m6A functions that may affect DON infection.
Project description:The intestinal epithelial cell lines 1 and J2 (IPEC-1, IPEC-J2) - spontaneously immortalised cell lines from the porcine intestine - are important tools for studying intestinal function. Microarrays (GeneChip Porcine Genome Array) were used to compare the expression pattern at basal in vitro conditions. Expression analyses complemented by morphological, functional and biochemical analyses revealed that IPEC-J2 is a morphologically and functionally more differentiated cell line in comparison to IPEC-1. In addition, IPEC-2 cells are a preferential tool for in vitro studies with the focus on metabolism.
Project description:To gain a more complete understanding of how porcine cathelicidin PR-39 influence the porcine intestinal epithelial cells, we profiled gene expression patterns in IPEC-J2 cell line in the presence or the absence of PR-39.
Project description:To understand the immunomodulatory effects of deoxyshikonin (a natural derivative of well-known Chinese medicine shikonin) on porcine intestinal epithelium cells, the transcriptome analysis was performed to explore the transcriptional profile of porcine IPEC-J2 cells after deoxyshikonin treatment.
Project description:The objective of this study is to investigate to what extent gene expression data of dietary interventions generated in IPEC-J2 in vitro model overlap with in vivo data. Gene expression was recorded in IPEC-J2 cells upon exposure to three different dietary interventions commonly used in livestock. In a further step, we compared the results with mucosal gene expression responses, as measured in animals exposed to the same compounds via the diet. As compounds we used zinc oxide, rye and the antibiotic amoxicillin. The GEO accession numbers of the in vivo datasets are provided in the paper "Enrichment of in vivo determined transcription data from dietary intervention studies with in vitro data provides improved insight into gene regulation mechanisms" (submitted to "Genes and Nutrition").
Project description:Marine algae convert a substantial fraction of the carbon dioxide they fix into various polysaccharides. Bacteria specialized on the remineralization of these polysaccharides often feature genomic clusters, termed polysaccharide utilization loci (PULs). Such PULs are often prevalent in, but not limited to, marine Flavobacteriia. Since knowledge on extant PUL diversity is sparse, we sequenced the genomes of 53 North Sea Flavobacteriia. We obtained 400 PULs, suggesting usage of a large array of polysaccharides, including laminarin, α- and β-mannans, fucose-, xylose-, galactose-, rhamnose- and arabinose-containing substrates, pectins, and chitins. Many of the PULs were novel, some indicating substrates that have rarely been described in marine environments. PUL repertoires of isolates often differed significantly within genera, corroborating ecological niche-associated glycan partitioning. Polysaccharide uptake in Flavobacteriia is mediated by SusCD. Respective protein trees revealed clustering according to polysaccharide specificities. Analysis of SusCD expression in multiyear phytoplankton bloom-associated metaproteomes indicated changes in microbial utilization of glucan, ß-mannan and sulfated xylan, suggesting that distinct substrates are temporarily abundant.
2018-08-16 | PXD008238 | Pride
Project description:Transcriptome sequencing between E. coli F18-induced IPEC-J2 and normal IPEC-J2 cells