We investigated a large outbreak of SARS-CoV-2 infections among passengers and crew members (60 cases in 132 persons) on a cruise ship sailing for 7 days on rivers in the Netherlands. Whole-genome analyses suggested a single or limited number of viral introductions consistent with the epidemiologic course of infections. Although some precautionary measures were taken, no social distancing was exercised, and air circulation and ventilation were suboptimal. The most plausible explanation for introduction of the virus is by persons (crew members and 2 passengers) infected during a previous cruise, in which a case of COVID-19 had occurred. The crew was insufficiently prepared on how to handle the situation, and efforts to contact public health authorities was inadequate. We recommend installing clear handling protocols, direct contacts with public health organizations, training of crew members to recognize outbreaks, and awareness of air quality on river-cruise ships, as is customary for most seafaring cruises.