AUTHOR=Bongiorno Dafne , Bivona Dalida A. , Cicino Claudia , Trecarichi Enrico M. , Russo Alessandro , Marascio Nadia , Mezzatesta Maria Lina , Musso Nicolò , Privitera Grete F. , Quirino Angela , Scarlata Giuseppe G. M. , Matera Giovanni , Torti Carlo , Stefani Stefania 

TITLE=Omic insights into various ceftazidime-avibactam-resistant Klebsiella pneumoniae isolates from two southern Italian regions

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=12

YEAR=2023

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2022.1010979

DOI=10.3389/fcimb.2022.1010979

ISSN=2235-2988

ABSTRACT=<p>Ceftazidime-avibactam (CZA) is one of the best therapeutic options available for infections caused by <italic>Klebsiella pneumoniae</italic> carbapenemase (KPC)-producing bacteria. However, sporadic reports of CZA-resistant strains have been rapidly increasing in patients. Herein, we provide detailed case reports of the emergence of ceftazidime-avibactam resistance to identify their resistome and virulome using genomic molecular approaches. Sixteen isolates were collected from 13 patients at three hospitals in Catania and Catanzaro (Italy) between 2020-2021. Antimicrobial susceptibility was determined by broth microdiluition. The samples included in study were analyzed for resistome, virulome and Sequence Type (ST) using Whole Genome Sequencing (WGS). All strains were resistant to ceftazidime/avibactam, ciprofloxacin, extended-spectrum cephalosporins and aztreonam, 13/16 to meropenem, 8/16 to colistin and 7/16 to fosfomycin; 15/16 were susceptible to meropenem/vaborbactam; all strains were susceptible to cefiderocol. Molecular analysis showed circulation of three major clones: ST101, ST307 and ST512. In 10/16 strains, we found a <italic>bla</italic><sub>KPC-3</sub> gene; in 6/16 strains, four different <italic>bla</italic><sub>KPC</sub> variants (<italic>bla</italic><sub>KPC28-31-34-50</sub>) were detected. A plethora of other beta-lactam genes (<italic>bla</italic><sub>SHV28-45-55-100-106-187-205-212</sub>, <italic>bla</italic><sub>OXA1-9-48</sub>, <italic>bla</italic><sub>TEM-181</sub> and <italic>bla</italic><sub>CTX-M-15</sub>) was observed; <italic>bla</italic><sub>OXA-9</sub> was found in ST307 and ST512, instead <italic>bla</italic><sub>OXA48</sub> in one out four ST101 strains. With regard to membrane permeability, <italic>omp</italic>K35 and <italic>omp</italic>K36 harbored frameshift mutations in 15/16 strains; analysis of <italic>omp</italic>K37 gene revealed that all strains harbored a non-functional protein and carry wild-type PBP3. There is an urgent need to characterize the mechanisms underlying carbapenem resistance and the intrinsic bacterial factors that facilitate the rapid emergence of resistance. Furthermore, it is becoming increasingly important to explore feasible methods for accurate detection of different KPC enzymes.</p>