All bacteria were grown anaerobically (85% N2, 10% CO2, 5% H2) at 37 °C in a modified Brain Heart Infusion (BHIS-YE) medium. This medium contains 37 g/L Brain Heart Infusion (BD Biosciences) supplemented with 5 mg/L of hemin, 2 mg/L of vitamin K1, and 5 g/L of yeast extract. Solid agar plates were prepared by combining Brain Heart Infusion, yeast extract, and 15 g/L agar and then autoclaving for 30 min at 121 °C. Once cooled, the agar was supplemented with hemin (5 μg/mL) (MilliporeSigma) and vitamin K1 (1 μg/mL) (Alfa Aesar) and plates were poured. For liquid media only (BHISG-YE), dextrose was added to a final concentration of 100 mM to promote biofilm formation and MgSO4 and CaCl2 was added to a final concentration of 1 mM to promote phage adhesion [52 (link)]. All components were dissolved in MilliQ water and filter-sterilized. Overnight cultures used in all experiments were grown statically in 6-well plates with 3-5 mL of BHISG-YE media.
Columbia blood agar plates were used to plate in vivo samples. This medium consists of 35 g/L of Columbia Broth Powder (BD Difco™) dissolved in water and autoclaved 30 min at 121 °C. Once cool, and before pouring plates, defibrinated sheep blood (Hemostat Laboratories) was added to the media at a final volume of 5% (v/v), and supplemented with hemin (5 μg/mL) (MilliporeSigma) and vitamin K1 (1 μg/mL) (Alfa Aesar).
Columbia blood agar plates were used to plate in vivo samples. This medium consists of 35 g/L of Columbia Broth Powder (BD Difco™) dissolved in water and autoclaved 30 min at 121 °C. Once cool, and before pouring plates, defibrinated sheep blood (Hemostat Laboratories) was added to the media at a final volume of 5% (v/v), and supplemented with hemin (5 μg/mL) (MilliporeSigma) and vitamin K1 (1 μg/mL) (Alfa Aesar).