The requirement for a low pH-dependent step during feline calicivirus (FCV) entry into Crandell-Reese feline kidney cells was investigated. Chloroquine, a lysosomotropic agent that prevents acidification of intracellular vesicles, inhibited the production of infectious virus when present during adsorption and the initial stages of FCV replication, but had little effect when added after 2 h post infection. The effect of chloroquine was reversible, allowing the virus growth curve to proceed when removed from the culture. In the presence of chloroquine small amounts of viral RNA were detected at 4, 6, and 8 h post infection, compared to untreated infected cells. These results suggest that entry of feline calicivirus into cells requires a low pH-dependent step.