This study determined the loading impacts of wood-based biochar on the eukaryotic community in three different soils (brown sandy loam—BSL, red loam—RL and a black clay loam—BCL) using a pot trial conducted over 10 months. Soil analysis and 18S rRNA gene sequencing performed using the Illumina MiSeq platform was carried out to evaluate the changes in eukaryotic community composition in relation to different added amounts of biochar. It was found that biochar addition had a negligible effect on diversity parameters in the brown sandy loam Kurosol (BSL) and red loam Dermosol (RL) soils. There were, however, significant changes in eukaryotic community composition of these biochar amended soils. These changes were most discernible in the lighter (low clay content) BSL soil for the fungal communities (F = 3.0106, p = 0.0003) present and also when total eukaryotes were considered (F = 2.3907, p = 0.0002). In this respect Glomeromycota seem to be slightly promoted in the lighter BSL soils, which might be due to increased soil porosity and soil chemical fertility. Clay rich BCL soil community structure correlated to a greater degree with soil chemistry influenced by biochar addition. The results showed that soil microeukaryotes were affected by short term carbon amendment, though to a limited extent. The limited effect of biochar loading rates on the soil microbiology could be due to the short incubation period, the lack of added fertiliser nutrients, and also the inherent stability of the soil eukaryotic community. The data suggested the impacts that were observed however included important plant symbiotic organisms. The results also imply biochar applications at different loading levels have differential effects on soil microeurokaryotes in relation to soil properties in particular clay content.