Human seroprevalence of Flavivirus was determined by hemagglutination inhibition tests on 479 sera from Misiones and 49 from Corrientes provinces. Paraná and Uruguay river bank communities from Argentina and neighbouring countries carry out frequent traffic across the rivers. With the aim of searching for a possible introduction of Dengue virus from Brasil or/and Paraguay, reactivity among people from Paraná and Uruguay river communities was compared with those from mountain communities. Two sera from Ituzaingó (Corrientes Province) were positive for Dengue 2. In Misiones, 3 sera from Oberá and 2 from Montecarlo were reactive for Dengue 2 and 1 serum from Puerto Iguazú was reactive for Dengue 1. Seroprevalence among the river population was significatively higher than among the mountain population. Likewise, populations on Paraná river showed more positive sera than those on Uruguay river; 54% of the samples possessed titers for SLE virus higher than for Dengue or Yellow fever. Anti-alphavirus (EEE and WEE) antibodies tested in sera from Misiones people showed a complementary distribution pattern to flavivirus. Seroprevalence of anti-alphavirus antibodies was higher in the mountain than in the river populations.