2.5. One step growth assays
Mid-exponential host bacterial cultures of V. parahaemolyticus were
adjusted to a 1 O.D. at 600 nm (corresponding to a cell density of
109 CFU mL−1). Ten microliters of the phage suspension was added to
10 mL of the bacterial culture in order to have a multiplicity of infection
(MOI) of 0.001. The phage was allowed to adsorb for 5 min at 25 °C,
without shaking. The mixture was centrifuged at 13,000 rpm for
5 min, the pellet was re-suspended in 10 mL of TSB at 37 °C and was
then serially diluted to 10–4. Samples (1 mL) were taken at 10–
20 min intervals and subjected to phage titration by the double-layer
agar method. Three independent assays were done.