Three different methods were used for estimating the basic reproductive number (R0) from data on 110 outbreaks of highly pathogenic avian influenza (HPAI) subtype H5N1 that occurred in village poultry in Romania, 12 May to 6 June 2006. We assumed a village-level infectious period of 7 days. The methods applied were GIS-based identification of nearest infectious neighbour (based on either Euclidean or road distance), the method of epidemic doubling time, and a susceptible–infectious (SI) modelling approach. In general, the estimated basic reproductive numbers were consistent: 2·14, 1·95, 2·68 and 2·21, respectively. Although the true basic reproductive number in this epidemic is unknown, results suggest that the use of a range of methods might be useful for characterizing epidemics of infectious diseases. Once the basic reproductive number has been estimated, better control strategies and targeted surveillance programmes can be designed.