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The Illumina Miseq System: A Critical Tool For Evaluating Host-Pathogen Interactions And Identifying Genomic Markers For Livestock Disease C


<p>Our long-term goals are to elucidate the mechanisms involved in host-pathogen interactions, identify mechanisms by which PDA prevent pathogenesis, develop early detection methods for mycobacterial diseases, and identify alternative methods to improve food safety. The Illumina® MiSeq® platform will allow us to test the overall hypotheses that (1) host-pathogen interactions induce changes in gene expression, which underlie immuno-pathogenesis in livestock, and that (2) PDA alter pathogenesis by modulating gene expression in the pathogens. </p><p>Objective 1: To determine the molecular pathways by which PDA reduce adhesion and/or invasion of S. uberis at the pathogen and host cell level.</p><p> Objective 2: To further investigate the bovine host transcriptional response to infection with Mycobacterium avium subspecies paratuberculosis (MAP) enabling improved diagnostic and prognostic biomarkers of Johne's disease (JD). </p><p>Objective 3: A) To determine the efficacy of orally supplemented trans-cinnamaldehyde (TC) and carvacrol (CR) for controlling Clostridium difficile-associated disease CDAD in a mouse model. B) To determine the effect of TC and CR supplementation on the intestinal microbiome of mice. </p><p>Objective 4: To characterize the molecular mechanisms involved in the colonization and survival of Escherichia coli O157:H7 on in-shell hazelnuts.</p>

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Several projects in the investigators' laboratories will benefit from use of the MiSeq.To determine the host-pathogen interaction in mastitis, primary mammary epithelial cells will be infected with S. aureus and RNA extracted. RNA will be sequenced using the MiSeq to determine differential gene expression. Additionally, the effects of plant-derived antimicrobials on S. aureus gene expression will be determined. To fill gaps in knowledge for the host response to Johne's disease, blood samples will be taken from infected and non-infected cattle and sequenced using the MiSeq. To determine the efficacy of plant-derived antimicrobials in preventing C. difficle associated disease, mice will be infected and treated with plant molecules. The pathogen will then be sequenced usign the miSeq. To characterize the molecular mechanisms involved in colonization and survival of E. Coli O157:h7 on in-shell hazelnuts, RNA will be collected from infected nuts and E. Coli and differential gene expression determined.

Govoni, Kristen E.
University of Connecticut
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