Background – The first records of Protosteloid Amoebae in the Democratic Republic of the Congo are discussed in the present paper. This survey on Protosteloid Amoebae is the first from Central Africa; the previous records for the African continent were restricted to Egypt, Kenya, Malawi, Tanzania and Uganda. Methods – Aerial litter samples, collected in 2010 during the “Boyekoli Ebale Congo” expedition in the Congo River basin between the cities of Kisangani and Bumba, were put into culture on wMY medium, a weak malt yeast nutrient agar medium. Results – The aerial litter cultures revealed 23 species representing 70% of the total number of species described worldwide. Two of these taxa, Schizoplasmodiopsis reticulata and Schizoplasmodium seychellarum, are new records for the African continent. The isolate LHI05 was observed for the first time on a substrate collected outside Hawai’i. In addition, 5 unknown taxa were observed. A selection of micrographs is presented of the new recorded species, the unknown taxa and all their related species observed in this study. Conclusion – The high species diversity observed on a limited number of samples suggests that the investigated region is, together with Hawai’i, one of the world’s tropical hotspots for Protosteloid Amoebae.