F. alocis ATCC 38596 was cultured in brain heart infusion (BHI) broth supplemented 5 mg/mL yeast extract, L-cysteine (0.05%), and arginine (0.05%) for 7 days anaerobically at 37°C as previously described (20 (link), 37 (link)). Serum opsonization was performed by incubating F. alocis at 37°C for 20 min in 10% normal human serum (Complement Technology, Inc., Tyler, TX, USA). Heat-killed F. alocis was generated by incubation at 90°C for 60 min. Non-viability was confirmed by incubation in culture media at the same conditions used for the live organism.
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