The emergence of high-risk clones of multidrug-resistant (MDR) bacteria in aquatic environments has generated an important public health problem, creating an urgent need to strengthen surveillance. This study reports the occurrence of clinically significant MDR Enterobacteriaceae and non-fermentative bacteria carrying carbapenemases (KPC-2), extended-spectrum β-lactamases (CTX-M) and plasmid-mediated quinolone resistance (PMQR) genes in urban lakes and reservoirs, in Southeastern Brazil. In this regard, the detection of hospital-associated lineages of KPC-2-producing Klebsiella pneumoniae belonging to the international clonal complex CC258 (ST11) and CTX-M-15-producing Escherichia coli belonging to the international CC10 (ST617), in an urban lake, is reported for the first time. Whole genome sequencing (WGS) analysis of KPC-2-producing K. pneumoniae ST11 revealed that blaKPC-2 gene was carried by an IncN plasmid on a Tn4401b element. This study support that aquatic environments with public access can act as reservoirs of clinically important MDR bacteria, constituting a potential risk to human and animal health. On the other hand, the detection of high-risk clones highlights the extra-hospital spread of clinically significant bacteria into urban aquatic environments. Display omitted •Presence of MDR bacteria in public lakes and reservoirs has been investigated.•Identification of high-risk clones of K. pneumoniae and E. coli is highlighted.•The environmental dissemination of hospital-associated lineages has been discussed.