Assessment of the Presence of Free-Living Amoebae in Soil Samples from the Northwest Region of Spain Using Culture and Molecular Assays.

Free-living amoebae (FLA) such as spp., , , spp., spp., and are naturally widespread in the environment, causing rare but fatal and debilitating infections in humans. In the present study, a total of 87 soil samples were collected from four provinces in the autonomous community of Castilla y León, Spain. These samples were collected in three different seasons during 2022-2023 (t1-t3) and were analysed by culture and molecular techniques (conventional PCR/sanger sequencing and qPCR). The obtained data revealed that the genus and the species were the most prevalent FLA. Furthermore, other genera/species of FLA were identified in the tested soil sources, suggesting a rich microbial biodiversity in Castilla y León soils. In addition, this study provides an important basis for future research on the ecology of these organisms and their potential impact on public health and the environment.