Introduction: Rabies is a viral disease with a worldwide distribution. Most often transmitted via domestic animals, the virus is also maintained and transmitted through sylvatic reservoirs. In South America, bats are the main vectors of the disease. In French Guiana, information on the distribution and circulation of the virus remains fragmentary. In 2008, a man died of rabies and viral genotyping revealed that it was of bat origin. Objectives: Our first objective was to determine the prevalence of rabies in bats in French Guiana. Then, we focused on two populations of vampire bats (Desmodus rotundus) to monitor their serological status over time. Materials and method: Bats were collected with nests from 2006 to 2014 in several landscapes. Individuals from two vampire bat colonies were also marked for capture-mark-recapture studies. Blood samples were collected for serological surveys and molecular investigations. A total of 996 sera from 29 species were tested. Results The overall rate of seropositivity was 10.1%, with important interspecific variations. Only one vampire was found positive for the presence of the virus. The monitoring of 2 vampire bat colonies revealed a high variation of seroprevalence over time. We also got evidence of changes for serological status for some animals on a one-year period. These results suggest that rabies virus actively circulates in the bat populations in French Guiana.  Conclusions: Longitudinal studies, based on the monitoring of the serological status and presence of the virus, associated to ecological and population genetics studies, have to be pursued to better understand its circulation in the D. rotundus communities.