The ancient lakes Mikri Prespa and Megali Prespa are located in SE Europe at the transnational triangle and are globally recognized for their ecological significance. They host hundreds of flora and fauna species, and numerous types of habitat of conservational interest. They also provide a variety of ecosystem services. Over the last few decades, the two lakes have been interconnected through a surface water channel. In an attempt to explore whether such a management practice might alter the ecological properties of the two lakes, we investigated a series of community metrics for phytoplankton by emphasizing cyanobacteria. Our results demonstrate that the cyanobacterial metacommunity structure was affected by directional hydrological connectivity and high dispersal rates, and to a lesser extent, by cyanobacterial species sorting. Cyanobacterial alpha diversity was twofold in the shallow upstream Lake Mikri Prespa (Simpson index average value: 0.70) in comparison to downstream Lake Megali Prespa (Simpson index average value: 0.37). The cyanobacterial assemblage of the latter was only a strict subset of that in Mikri Prespa. Similarly, beta diversity components clearly showed a homogenization of cyanobacteria, supporting the hypothesis that water flow enhances fluvial translocation of potentially toxic and bloom-forming cyanobacteria. Degrading of the water quality in the Lake Megali Prespa in anticipation of improving that of the Lake Mikri Prespa is an issue of great concern for the Prespa lakes’ protection and conservation.