1a.Objectives (from AD-416):
To determine gene expression changes in monocytes/neutrophils isolated during the progression of a mammary gland infection in dairy cattle.
1b.Approach (from AD-416):
Lactating dairy cows will be infected with S. uberis. At various time points cells will be isolated from milk and sorted by cell type. Monocytes and neutrophils will be isolated and stored for subsequent RNA analysis. RNA from cells will be isolated and used to perform large-scale RNA sequencing. This data would be analyzed using current bioinformatics software designed by our collaborator. This data will become a part of a public database modeling bovine innate immunity during infections.
Cows were infected with S. uberis and at various time points milk was collected and CD14+ monocytes were isolated from milk and other cells. Monocytes were processed to purifiy DNA, RNA, and proteins and each was separated and stored for various assays. The DNA was sent to a researcher at the Beltsville Agricultural Research Center (BARC). The RNA, both mRNA and miRNA, was sent to the Iowa State University Sequencing Facility. Sequencing of mRNA was completed last year and data was shipped to researchers at Teagasc for analysis. The sequencing facility finished the analysis of the miRNA. All data has been transferred to our collaborators in Ireland for processing. We anticipate the processing of data will be complete in early FY 2014. Data interpretation is complete and the manuscript will be submitted to a peer-reviewed journal shortly.