In South America, the Amazon basin is considered an endemic area of hepatitis B virus (HBV) infection. However, epidemiological studies with vulnerable groups are scarce. Female sex workers (FSWs) are highly vulnerable to sexually transmitted infections due to a combination of their sexual behavior and socio-economic conditions. Thus, this study investigated the prevalence of HBV infections and HBV–hepatitis C virus, HBV–hepatitis D virus, HBV–HIV, and HBV–human T-lymphotropic virus co-infections among FSWs in the Marajó Archipelago, northern Brazil, as well as identifying the HBV genotypes circulating in this population. A total of 153 FSWs in 5 towns and 18 riverside communities were included in the study. The HBV infection and co-infections were diagnosed by enzyme-linked immunosorbent assay and real-time polymerase chain reaction. The HBV genotypes were detected by sequencing and were then analyzed phylogenetically. Most of the FSWs surveyed were single, young, heterosexual, and born locally, with low levels of education. Overall, 21 (13.7%) had been exposed to HBV, and HBV-DNA was detected in 13 (8.5%). Genotypes A (69.2%), D (23.1%), and F (7.7%) were detected. Seven cases of co-infections with other viruses were detected. These findings indicate a clear need for urgent measures to control the spread of HBV and other pathogens, and to promote the health of the local FSWs.