Source: UNIV OF CONNECTICUT submitted to
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
Funding Source
Reporting Frequency
Accession No.
Grant No.
Project No.
Proposal No.
Multistate No.
Program Code
Project Start Date
Feb 15, 2015
Project End Date
Feb 14, 2017
Grant Year
Project Director
Govoni, K.
Recipient Organization
Performing Department
Animal Science
Non Technical Summary
A thorough understanding of the mechanisms of pathogenesis and host-pathogen interactions is critical for the development of innovative approaches and tools in preventing disease in livestock. In addition, there is a need for alternative treatment therapies to reduce the use of antibiotics in livestock. Genome sequencing, relative mRNA expression analysis and identification of epigenetic modifications are cutting edge areas of research for identify mechanisms of pathogenesis and potential targets for novel treatments of disease. The purchase of the Illumina MiSeq, a sequencing machine, will allow the investigators to sequence pathogens involved in livestock diseases, such as mastitis and Johne's disease and pathogens involved in food borne illnesses. Sequencing data can be used to identify novel genes and pathways involved in infection. This work is important to identify novel methods to prevent and treat diseases in livestock, thereby improving quality of life for animals and efficiency of production. This will directly benefit consumers by reducing the cost of products such as meat and milk.
Animal Health Component
Research Effort Categories

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
Goals / Objectives
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. 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. 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). 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. Objective 4: To characterize the molecular mechanisms involved in the colonization and survival of Escherichia coli O157:H7 on in-shell hazelnuts.
Project Methods
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 reponse to Johne's disease, blood samples will be taken from infected and non-infected cattle and sequenced using the MiSeq. To determine the efficiacy 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 mechansims 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 geneexpression determined.

Progress 02/15/15 to 02/14/17

Target Audience:Use of the MiSeq and data generated have targeted scientists and educators in the field of microbiology and animal physiology. Additionally, graduate and undergraduate students were trained in techniques and knowledge of next generation sequencing. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Graduate students and undergraduate students were trained in cell culture, RNA extraction and library preparation and sequencing. Additionally, they were trained in data analysis and interpretation, manuscript preparation and presentation of data. How have the results been disseminated to communities of interest?Initiall findings were presented at the American Society of Animal Science meeting and are currently in preparation for manuscript submission. Findings were also presented to faculty, staff and students at the University of Connecticut. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

What was accomplished under these goals? We have demonstrated that the host-pathogen interaction between Staphylococcus Aureus and bovine mammary epithelial cells alters gene expression in both the host and the pathogen. Additionally, these changes are associated with changes in cell function, viability and adhesion. Plant-derived antimicrobials reduced adhesion and invasion of S. Aureus to mammary epithelial cells in culture and these changes are associate with changes in cell adhesion gene expression in the pathogen. These findings are currently in preparation for publication.


  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2015 Citation: Valley, EV, Jaganathan, D, Venkitanarayanan, K, Kazmer, GW, Kuo, L, Wang, YB, and Govoni, KE. 2015. Effects of plant derived compounds on Staphylococcus aureus infection of primary bovine mammary epithelial cells. J. Anim. Sci. Vol. 93, Suppl. s3.

Progress 02/15/15 to 02/14/16

Target Audience:The experiments that will utlize the Illumina MiSeq Equipment target a scietific audience including scientists, faculty, postdocs, graduate students and undergraduates. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This equipment and the proposed exerperiments have provided training for graduate and postdocs in whole transcriptome sequencing analysis. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?Additional sequencing analysis will be completed using the MiSeq to meet the remaining objective in the proposal.

What was accomplished under these goals? Use of the Illumina Mi-Seq analyzer, we have sequenced the whole transcriptome of staphylococcus aureus in the absence and presence of plant-derived compounds trans-cinnemaldehyde (TC) and eugenol (EG). Treatment of SA with TC resulted in 110 differentially expressed genes of which 3 were down-regulated greater than 2-fold and 10 were up-regulated greater than 2-fold. Treatment of SA with EG resulted in 222 differentially expressed genes of which 17 were down-regulated by greater than 2-fold and 13 were up-regulated greater than 2-fold. To gain a better understanding of the host-pathogen interaction during mastitis infection, we developed an in vitro model in which MEC were infected with SA and then RNA was isolated from both the MEC and SA for whole transcriptome analysis. RNA-Seq analysis using the MiSeq determined that 706 genes in SA are differentially expressed when exposed to MEC. Currently analysis is in progress to identify the functions and pathways in which these genes are involved.