Characterization of Klebsiella pneumoniae Isolates Resistant to Cefiderocol from Hospitals and Outpatient Settings in Croatia

Background/Objectives: We conducted this study to evaluate the genotypic and phenotypic profiles of carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates, exhibiting resistance to cefiderocol (FDC), focusing on antibiotic susceptibility, beta-lactamase production, the genetic environment of blaCARB and blaESBL genes and molecular epidemiology. FDC is now a last-line antibiotic for severe infections due to CRKP. Methods: Susceptibility to a wide range of antibiotics was determined by the disk diffusion and broth microdilution method. Carbapenemases were screened by a modified Hodge test while carbapenem hydrolysis was investigated using mCIM and eCIM tests. The screening for beta-lactamase and fluoroquinolone cluster resistance genes was carried out by PCR. Plasmids were characterized by PCR-based replicon typing (PBRT). An inter-array genotyping CarbaResist test and whole genome sequencing (WGS) were applied on selected isolates. Results: All of the 31 isolates studied exhibited high-level resistance to amoxicillin-clavulanate, piperacillin-tazobactam, cefuroxime, expanded-spectrum cephalosporins (ESC), cefepime, ceftolozan-tazobactam and ciprofloxacin and the majority to gentamicin, and amikacin. Colistin and ceftazidime-avibactam preserved activity against 71% and 87% of the isolates, respectively. The combined disk method with clavulanic acid was positive in all but one isolate, indicating the production of an ESBL. Twenty-eight isolates carried one single carbapenemase-encoding gene, whereas three harbored double blaCARB genes. Among the studied isolates, 61% carried blaOXA-48, 29% blaKPC and 12.9% blaNDM genes. The inter-array genotyping CarbaResist test and WGS identified additional aminoglycoside-, sulphonamide- and trimethoprim-resistance genes. Conclusion: To our knowledge, this is the first study on FDC resistance in Croatia. The diffusion of FDC-resistant isolates was detected in both hospital and outpatient settings, emphasizing the need for a "One Health" approach.