The investigation of New York African Burial Ground (NYABG) samples has allowed researchers at Howard University to reconstruct the bacterial community presence of burial soil of Africans who lived during slavery. Our analyses detected two distinct profiles in our NYABG samples: 1) an environmental signature providing details about the wetland, marshy environment of 17th Century New York, and 2) a human microbiome signature similar to that of living or recently deceased humans reported in the current scientific literature. This research sets a precedent for the successful identification of human presence in 400-year old buried soil. It helps us to learn about the quality of life of NYABG inhabitants (detection of infectious diseases) and the change in microbiome diversity over time (due to diet and societal lifestyle changes). Finally, this study demonstrates the potential to explore burial soil for human DNA of historical populations without disturbing their skeletal remains. This method may be helpful in historical, forensic, anthropological, molecular, environmental, and archaeological studies.