Rickettsia are obligate intracellular bacteria often associated with ticks and best known for causing human diseases (rickettsiosis), including typhus fever and sporadic cases of serious infection. in this study, we conducted a large survey of ticks in french Guiana to understand the overall diversity of Rickettsia in this remote area largely covered by dense rainforests. Out of 819 individuals (22 tick species in six genera), 252 (30.8%) samples were positive for Rickettsia infection. Multilocus typing and phylogenetic analysis identified 19 Rickettsia genotypes, but none was 100% identical to already known Rickettsia species or strains. Among these 19 genotypes, we identified two validated Rickettsia species, Rickettsia amblyommatis (spotted fever group) and Rickettsia bellii (bellii group), and characterized a novel and divergent Rickettsia phylogenetic group, the guiana group. While some tick hosts of these Rickettsia genotypes are among the most common ticks to bite humans in french Guiana, their potential pathogenicity remains entirely unknown. However, we found a strong association between Rickettsia genotypes and their host tick species, suggesting that most of these Rickettsia genotypes may be nonpathogenic forms maintained through transovarial transmission. Members of the Rickettsia genus are obligate intracellular bacteria of eukaryotes 1-3. The best known Rickettsia species are major human pathogens that include the etiological agents of the epidemic typhus, R. prowazekii, the Rocky Mountain spotted fever, R. rickettsii, and the flea-borne spotted fever, R. felis 1,4,5. Most of these pathogenic Rickettsia species have a zoonotic life cycle and are transmitted by blood-feeding arthropods such as ticks, mites, lice and fleas, which commonly serve as ecological bridges for transmission from wildlife to humans and domestic animals 1,4,5. However, members of the Rickettsia genus are actually more widespread than previously recognized: ecological surveys are uncovering substantial Rickettsia diversity associated with blood-feeding arthropods but also with non-blood-feeding arthropods, protozoa, algae and plants 1-3. There are currently more than 30 recognized Rickettsia species but the advent of multilocus sequence typing (MLST) and molecular phylogenetics has recently led to the description of several new putative species 1,2,4,6,7. Historically, Rickettsia were classified into a few major groups based on serological characteristics, but subsequent DNA sequencing led to classification of at least 10 distinct phylogenetic groups 1,2,6. Three Rickettsia groups-the spotted fever group, the typhus group and the transitional group-are the subject of intensive study since they all include major pathogenic species and are commonly found in blood-feeding arthropods 1. In addition, another Rickettsia group widely found in ticks but also in many other arthropods, the bellii group, is also commonly studied. This group is thought to be basal to the three other major Rickettsia groups and is largely composed of nonpath...