The Atlantic Forest Biome (AFB) creates an ideal environment for the proliferation of vector mosquitoes, such as Haemagogus and Sabethes species, which transmit the Yellow Fever virus (YFV) to both human and non-human primates (NHP) (particularly Alouatta sp. and Callithrix sp.). From 2016 to 2020, 748 fatal cases of YF in humans and 1,763 in NHPs were reported in this biome, following several years free from the disease. This underscores the imminent risk posed by the YFV. In this study, we examined the spatiotemporal distribution patterns of YF cases in both NHPs and humans across the entire AFB during the outbreak period, using a generalized linear mixed regression model (GLMM) at the municipal level. Our analysis examined factors associated with the spread of YFV, including environmental characteristics, climate conditions, human vaccination coverage, and the presence of two additional YFV-affected NHP species. The occurrence of epizootics has been directly associated with natural forest formations and the presence of species within the Callithrix genus. Additionally, epizootics have been shown to be directly associated with human prevalence. Furthermore, human prevalence showed an inverse correlation with urban areas, temporary croplands, and savannah and grassland areas. Further analyses using Moran's Index to incorporate the neighborhoods of municipalities with cases in each studied host revealed additional variables, such as altitude, which showed a positive correlation. Additionally, the occurrence of the disease in both hosts exhibited a spatio-temporal distribution pattern. To effectively mitigate the spread of the virus, it is necessary to proactively expand vaccination coverage, refine NHP surveillance strategies, and enhance entomological surveillance in both natural and modified environments.