Leptospira and E. coli Cultivation Protocol

Pathogenic L. interrogans serovar Copenhageni strain FIOCRUZ L1-130 and a recently isolated pathogenic non-interrogans Leptospira strain LGVF02 from soil samples (Nathan Stone, Northern Arizona University, manuscript in preparation) were grown in liquid HAN media^{23 (link)} at 29 °C. Saprophytic L. biflexa serovar Patoc strain Patoc1 was cultured in EMJH medium (Difco, BD, Franklin Lakes, NJ). Solid media were prepared by supplementing with 1.2% noble agar (Difco). Where necessary, spectinomycin was added at 40 µg/mL. E. coli strains π1 and β2163^{24 (link)} were used for general cloning and as conjugation donor cells, respectively, and were grown in Luria–Bertani (LB, Difco) media. For E. coli strains π1, thymidine (dT, 0.3 mM, Sigma) was added to the media and for strain β2163, diaminopimelic acid (DAP, 0.3 mM, Sigma) was added.