In 2001, West Nile virus (WNV) activity was reported from 359 counties in 27 states and the District of Columbia (DC) to ArboNET, a web-based, surveillance data network maintained by 54 state and local public health agencies and CDC. This activity represented a marked increase from 2000, when WNV activity was reported from 138 counties in 12 states and DC. This report summarizes surveillance data for 2001, which indicate that 66 human illnesses were reported from 10 states and that widespread WNV activity in birds, horses, and mosquitoes extended into the midwestern United States and several southern states unaffected previously. The findings in this report underscore the need for public education, increased WNV surveillance aimed at early viral detection, and sustained, integrated mosquito-control activities.