Urban ecology has been an understudied topic in myxomycete research.For that reason, the present investigation aimed to generate data related to myxomycetes in urban areas of the Neotropical region.With an approach of two discrete experiments, one centered on ground-based information and one on air-based data, results showed that myxomycetes can be valuable organisms for microbial ecology assessments in urban centers.Data from these experiments showed that, using a simple and classical laboratory-based method of detection, the number of records seemed to be affected by the degree of urbanization, which also had an effect on pH values, but the number of species seemed to be more associated with site-specific characteristics.Airborne propagules of myxomycete dispersion, captured using substrates exposed to outdoor conditions, indicated that air currents may play a role on the distribution of myxomycetes in urban conditions, potentially affecting the process of ecological data generation.The results obtained herein are useful to demonstrate that myxomycetes can be studied in urban centers and that more systematic approaches could generate relevant data in the context of climate change, green cities, and urban biodiversity monitoring.