Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Gene transcription profiles showing early systemic response to E. coli (k2bh2) infection in bovine liver

ABSTRACT: We studied the response to infection and associated perturbations to the bovine liver’s normal function by examining gene transcription data from liver biopsies collected following an E. coli infection in the udder of primiparous dairy cows. This is the first study to examine gene transcription responses to systemic infection by the E. coli bacterium in dairy cows. First, we verified that the inoculation protocol resulted in systemic infection in the cows. This was done based on records on three clinical symptoms: body temperature and amount of E. coli bacteria and leukocytes in milk samples. Second, we examined gene transcription patterns underlying the clinical traits. Gene transcription levels at times of peak values for the clinical traits were estimated in the liver to study indications of an acute phase response to systemic E. coli infection in the cows. Finally, we compared gene transcription responses to E. coli infection and lipopolysaccaride (LPS) inoculation. Sixteen healthy, primiparous Danish Holstein-Friesian cows were challenged intramammarily with E. coli 4 to 6 weeks following parturition. Gene transcription levels for fifteen of the cows were used to study infection responses. We compared gene transcription levels before and after E. coli infection for all fourteen cows.

ORGANISM(S): Bos taurus

SUBMITTER: A.J. (Bart) Buitenhuis

PROVIDER: E-GEOD-25319 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Although the majority of previous work on campylobacteriosis has centered on the species Campylobacter jejuni, Campylobacter coli, the sister group to C. jejuni, is also a significant problem, but remains a much less studied organism. The purpose of this study was to develop and apply an expanded 16 locus MLST genotyping scheme to a large collection of C. coli isolates sampled from a wide range of host species, and to complete microarray comparative genomic hybridizations for these same strains, in order to: (1) determine whether host specific clones, genotypes, or clonal complexes are evident and (2) evaluate whether there are particular genes comprising the dispensable portion of the C. coli genome that are more commonly associated with certain host species. Genotyping and ClonalFrame analyses of the expanded MLST data suggest that (1) host preferred groups have tended to evolve in the diversification of C. coli, (2) this has happened repeatedly, at different times, throughout the evolutionary history of the species, and (3) recombination has played varying roles in the diversification of the different groups. Concomitant with the information on evolutionary history derived from the MLST data, the microarray data suggests that a combination of common ancestry in some cases and lateral gene transfer in others are behind a tendency for sets of genes to be common to isolates derived from particular hosts. Keywords: comparative genomic hybridization Combimatrix CustomArray™ 4X2K was used in this study. This array is divided into 4 sectors, each of which contains 2,240 in situ synthesized oligonucleotide probes (spots) with the same probe design and layout. Based on the sequence of Campylobacter coli strain RM2228, oligonucleotide probes were designed to have a similar annealing temperature of 56ºC and a length 35-40 bp. Two separate designs were used in this study; both included 100 control probes (20 negative controls with sequences from plant and phage, each with 5 replicate spots) as well as loci from the RM2228 genome. Because of the strict criteria for probe design, not all ORFs could be covered in this analysis. The first design included 1942 of the 1967 protein coding genes described in the unfinished sequence of C. coli strain RM2228. The second-generation design was based on genes that were not clearly present (loci with low intensity or no hybridization for at least one strain) in the hybridization results involving the first design. The second design included additional two or five probes, separated from one another in order to span the entire gene, for these 615 ambiguous loci, synthesized in situ to occupy the 2,240 independent microarray spots. Replicate microarrays were hybridized for every 65 strains tested in this study.

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Gene transcription profiles showing early systemic response to E. coli (k2bh2) infection in bovine liver

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