Samples from patients with suspected HAI were collected and transported to the laboratory following the general specimen collection and transport techniques [4]. A total of 32 P. aeruginosa isolates were analyzed using all of the following methods: conventional phenotypic methods, molecular tests, and whole-genome sequencing. Identification and susceptibility to antimicrobials were performed by automated Vitek-2 Systems (BioMérieux, Marcy-l’Étoile, France) using CLSI breakpoints for Carbapenems (MIC ≥ 8 µg/mL) [5]. Phenotypic studies included modified carbapenem inhibition test (mCIM) according to CLSI [5] and metallo-ß-lactamase (MBL) E-test (BioMérieux, Marcy-l’Étoile, France). Molecular analyses for detection of genes conferring resistance included Xpert Carba Test (Cepheid, Sunnyvale CA, USA) for detection of genes encoding carbapenemases KPC, VIM, IMP, NDM, OXA-48; AMR Flow-Chip hybridization (Master Diagnóstica, Granada, Spain) for detection of genes encoding extended-spectrum beta-lactamases SHV, CTX-M; class A carbapenemases GES, SME, KPC, IMI; class B carbapenemases SIM, GIM, SPM, NDM, VIM, IMP and class D carbapenemases OXA23-like, 24-like, 48-like, 51-like and 58-like; and detection of genes encoding carbapenemases IMP and VIM by Polimerase Chain Reaction (PCR) Laboratory-Developed Tests (LDT) using primers described elsewhere [6]. When required, DNA was extracted using MagNA Pure (Roche Diagnostics, Basel, Switzerland), quantified by Quantus (Promega, Madison WI, USA), and verified by Qiaxcell (QIAgen, Germantown MD, USA). The whole genome was prepared using the Nextera Flex library preparation standard protocol and paired-end fragment (151 bp) sequencing (MiSeq, Illumina Inc., San Diego CA, USA). A general pipeline for read quality (FastQC) [7], trimming (Trimmomatic) [8], genome assembly (Shovill) [9], genome annotation (Prokka) [10] and resistance-gene finder (ABRicate) [11] was performed using Galaxy Community Hub [12].