Seroepidemiological surveys are epidemiological studies carried out by the use of serum tests to detect infection; they can be applied to infections in general and to vaccine-preventable diseases in particular. Among other applications, seroepidemiological studies are useful for determining groups at risk for a specific disease; evaluating transmission mechanisms; and determining population groups who are critical in maintaining the transmission of infectious agents. We analysed the results of seroprevalence studies in Spain on vertically-transmitted non vaccine-preventable diseases (Toxoplasma gondii, Treponema pallidum, cytomegalovirus, herpes simplex virus type 2, and parvovirus human B19), blood-borne diseases (hepatitis C virus, human immunodeficiency virus and Trypanosoma cruzi) and emerging diseases (West Nile virus, Toscana virus, and lymphocytic choriomeningitis virus). We found a reduced seroprevalence against Toxoplasma gondii, Treponema pallidum, cytomegalovirus and human immunodeficiency virus in the Spanish indigenous population. The results found in immigrants reflect the situation of these diseases in their countries of origin and suggest they could have a substantial public health impact in Spain (Trypanosoma cruzi, in association with blood donation). We highlight the circulation in Spain of the West Nile virus and the importance of Toscana virus infection.