Location: Geospatial and Environmental Epidemiology Research Unit
Project Number: 6064-32000-001-007-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Aug 15, 2024
End Date: Aug 14, 2028
Objective:
In collaboration with GEERU and ADRU generate complete Anaplasma (A.). marginale genomes from geographically disparate regions of North America. Additionally develop and validate the methods to directly sequence A. marginale from low level A. marginale infections.
Approach:
Initially Anaplasma (A.) marginale strains will be expanded in cattle using existing stabilates. Blood will be collected, washed, and genomic DNA isolated for whole genome sequencing. Concurrently, the cooperator will develop and validate methods to isolate A. marginale infected erythrocytes, with cell sorting being the primary approach, however other strategies may be tested (gradient centrifugation, enrichment kits). Genomic DNA from A. marginale will be extracted and evaluated for quality and contamination with bovine DNA to determine suitability for whole genome sequencing. DNA extraction protocols will be optimized as needed. During development of these methods, the cooperator will reach out to collaborators in the field to establish sample processing protocols and coordinate the receipt of either blood for DNA samples for sequencing at GEERU.
