Optimization of in vitro gowth conditions and DNA extraction from Treponema phagedenis isolated from bovine digital dermatitis lesions

Abstract

Bovine digital dermatitis is a leading cause of lameness in the US dairy industry, and it also represents a significant welfare issue. Our group and others have demonstrated that, along with a number of other organisms, Treponema spp can be cultured from a large percentage of digital dermatitis lesions. As part of our ongoing effort to better understand the role that Treponema spp plays in this disease process, we are using a combination of culture-dependent and independent methods to better understand the physiology and metabolism of these bacteria. Treponema spp are fastidious and their growth in broth media has traditionally been difficult; thus; hindering the ability to grow sufficient organisms for DNA purification to be used in downstream sequencing or genetic modification. This study details the systematic evaluation of different culture conditions on the growth and replication of T. phagedenis isolated from digital dermatitis lesions of cattle.