The first outbreak of hantavirus pulmonary syndrome (HPS) in Central America was documented on the Azuero peninsula of Panama in late 1999 and 2000. Reverse transcriptase-polymerase chain reaction evidence implicated only Choclo virus in symptomatic HPS with a mortality rate of 20%, although two rodent-borne hantaviruses (Choclo virus and Calabazo virus) were identified in the peridomestic habitat. Neighborhood serosurveys around case households found seroprevalence rates as high as 30%, the highest in the Americas except for western Paraguay. We report here population-based serosurveys for 1,346 adults and children in four communities, three on the Azuero peninsula and one in adjacent central Panama. Overall seroprevalence ranged from 33.2% in a population engaged in farming and fishing on Isla de Cañas, to 16.3% and 21.2% in two mainland agricultural communities, to 3.1% in central Panama, with a modest male predominance of 1.2:1. Nine percent of children 4-10 years old were seropositive, and seroprevalence increased with age in all communities, with highest levels of 52% in those 41-50 years old cohort on Isla de Cañas. Univariate analysis identified correlations between seroprevalence and multiple agricultural and animal husbandry activities. However, stepwise logistic regression models identified only raising animals (cows, pigs, goats, poultry) and fishing as significant independent variables. Human infection with hantavirus on the Azuero peninsula, either with Choclo virus or combined with Calabazo virus, is frequent but rarely results in hospitalization due to respiratory illnesses resembling HPS.