To understand transmission of avian influenza A (H5N1) virus, we conducted a retrospective survey of poultry deaths and a seroepidemiologic investigation in a Cambodian village where a 28-year-old man was infected with H5N1 virus in March 2005. Poultry surveys were conducted within a 1-km radius of the patient's household. Forty-two household flocks were considered likely to have been infected from January through March 2005 because >60% of the flock died, case-fatality ratio was 100%, and both young and mature birds died within 1 to 2 days. Two sick chickens from a property adjacent to the patient's house tested positive for H5N1 on reverse transcription-PCR. Villagers were asked about poultry exposures in the past year and tested for H5N1 antibodies. Despite frequent, direct contact with poultry suspected of having H5N1 virus infection, none of 351 participants from 93 households had neutralizing antibodies to H5N1. H5N1 virus transmission from poultry to humans remains low in this setting.