To examine the seroprevalence of zoonotic visceral leishmaniasis (ZVL) among rodents in Azarshahr County, and to assess the possible association of infection among rodents in respect with transmission/prevalence of the disease among children, a survey was conducted during 2003-2004. Azarshahr County is an endemic region for leishmaniasis and this research is the first study in determining the host reservoirs in this county. In this survey, 265 rodents belonging to 7 genera/species were trapped alive. Anti-Leishmanial antibodies were detected through direct agglutination test (DAT), indirect fluorescent antibody tests (IFAT) and microscopic examination. Fourteen (5.3%) animals were shown to be seropositive, 27 (10.2%) provided relatively lower titers, and 224 (84.5%) turned out to be seronegative. Amastigotes of Leishmania were observed in 4 seropositive rodents including 1 Meriones persicus, 2 Cricetalus migratorius and 1 Mesocicetus auratus after dissection and parasitological examinations. Multiple analyses of PCR were used to reassure the identity of purified isolates and infected clinical samples. According to the results of this study, the isolates were identified as Leishmania infantum and those infected rodents are assumed to be potential host reserviors for visceral leishmaniasis in the region. This work is the first report of detecting L. infantum infection in Cricetalus migratorius, Meriones persicus and Mesocricetus auratus from Azarshahr of Iran.