Antimicrobial resistance is a public health concern affecting human, animal, and environmental health. Antimicrobial-resistant pathogens, such as Enterococcus spp., Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp., and Escherichia coli (ESKAPEE), the most important microorganisms involved in healthcare-related infections, can be excreted by patients into hospital effluent which then becomes a reservoir for these pathogens. In this context, we characterized and compared resistant microorganisms of the ESKAPEE group, isolated from hospital effluent and clinical samples from patients of the Pequeno Príncipe Hospital, located at Curitiba, Paraná. Out of 345 microorganisms isolated, 208 from the ESKAPEE group were obtained from hospital effluent and 18 were from clinical samples. Among them, K. pneumoniae was the most frequently identified microorganism in both clinical and environmental settings. The genomic context of the resistance of 52 single-clone isolates with satisfactory genome assembly parameters was analyzed. The identified carbapenem resistance genes were bla