The pollution of aquatic ecosystems is a big problem that has its impact on river sediments. In recent decades, an effective solution to this problem has been the application of bioremediation technologies. Nanoremediation is an innovative part of these technologies. We still know little about the efficiency of nanoparticles, especially nanodiamonds, in modelled conditions. The aim of the present study is to investigate the effect of nanodiamonds on the key parameters of modelled bioremediation of river sediments that are polluted with phenol, as well their effect on the structures and functions of microbial communities. An important indicative mechanism that was used is the application of fluorescent in situ hybridization for sediment microbial communities. The results of this study revealed the positive role of nanodiamonds that is associated with their intoxication with high concentrations of phenol. Readaptation was also found, in which the xenobiotic biodegradation potential evolved by increasing the relative proportions of non-culturable bacteria, namely Acinetobacter (at the 144th hour) and Pseudomonas (at the 214th hour). The results can help to find an effective solution to the question of how information from such precise molecular methods and the application of nanodiamonds can be translated into the accessible language of management and bioremediation technologies.