The application of recycled concrete aggregates (RCAs) has become increasingly popular for different types of structures, as presented in several studies. However, depending on the type of structure and the region, RCAs might have different properties. This study aims to investigate the application of RCAs of different origins for substructure layers of the cycle paths located in Central Europe, which was not analysed previously. Recycled aggregates from an airport, road overpass, and building demolition were tested according to European standards and used to produce concretes, in which compressive strength, density, water absorption, and frost resistance were tested. After 28 days, RCA concrete had compressive strengths from 5.9 to 17.3 MPa and frost resistance ratios close to 1.0. The concrete parameters indicate that RCAs might be used for the construction of cycle path substructural layers with the appropriate class of cement and W/C ratio. To meet the requirements of EN 12390-3 to achieve class C8/10, RCA concrete with CEM II B/V 32.5 should be used with a W/C ratio of 1. To meet the requirements of D-04.05.01v02, RCA concrete with CEM II B/V 32.5 and a W/C ratio smaller than 1.50 should be used. Applying recycled RCAs in various structures helps protect natural resources by reusing materials. However, the variability in RCA properties requires testing to guarantee quality.