Location: Animal Disease Research
Project Number: 2090-32000-043-003-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Aug 1, 2024
End Date: Jul 31, 2026
Objective:
The objective of this agreement is to develop a nutrient medium to support robust and sustained metabolic activity of A. marginale in the absence of host cells.
Approach:
Using an iterative process, we will identify the components of a basal medium (i.e., buffer with ion concentrations suitable for pathogen integrity and viability, and amino acids to support basal protein synthesis) that support metabolic activity of A. marginale in the absence of host cells. We will then build on the basal medium with identification of a carbon source and other nutrients such as iron, cholesterol and specific amino acids required by A. marginale to support sustained metabolic activity. Protein synthesis is used as a robust marker of metabolic activity and is measured by incorporation of S35-labeled cysteine and methionine into A. marginale proteins. The use of protein synthesis as a basis for the analysis allows for a measure of biomass generation via macromolecular de novo biosynthesis without requiring cell division.
