Abstract
Carbapenem-resistant Klebsiella pneumoniae are a major global threat in healthcare facilities. The propagation of carbapenem resistance determinants can occur through vertical transmission, with genetic elements being transmitted by the host bacterium, or by horizontal transmission, with the same genetic elements being transferred among distinct bacterial hosts. This work aimed to track carbapenem resistance transmission by K. pneumoniae in a healthcare facility. The study involved a polyphasic approach based on conjugation assays, resistance phenotype and genotype analyses, whole genome sequencing, and plasmid characterization by pulsed field gel electrophoresis and optical DNA mapping. Out of 40 K. pneumoniae clinical isolates recovered over two years, five were carbapenem- and multidrug-resistant and belonged to multilocus sequence type ST147. These isolates harboured the carbapenemase encoding blaKPC-3 gene, integrated in conjugative plasmids of 140 kbp or 55 kbp, belonging to replicon types incFIA/incFIIK or incN/incFIIK, respectively. The two distinct plasmids encoding the blaKPC-3 gene were associated with distinct genetic lineages, as confirmed by optical DNA mapping and whole genome sequence analyses. These results suggested vertical (bacterial strain-based) transmission of the carbapenem-resistance genetic elements. Determination of the mode of transmission of antibiotic resistance in healthcare facilities, only possible based on polyphasic approaches as described here, is essential to control resistance propagation.
Highlights
The discovery of antibiotics during the 20th century led to a decrease in deaths due to infectious diseases
From a group of 40 K. pneumoniae isolates recovered from patients over a two-year period in the studied hospital, five were carbapenem-resistant (KP1-388, KP2-448, KP2-465, KP1-080 and KP1-349), and were isolated from distinct biological samples of two patients, at different time points over a period of 18 months
The five isolates belonged to the same MLST group, ST147 (Table 1), based on whole genome sequence analysis, they could be divided into two groups, one comprising the patient 1 isolates (KP1-388, KP1-080 and KP1-349) and the other the patient 2 isolates (KP2-448 and KP2-465)
Summary
The discovery of antibiotics during the 20th century led to a decrease in deaths due to infectious diseases. The highly dynamic genome of K. pneumoniae explains the frequent acquisition of resistance to multiple antibiotics, often through conjugative horizontal gene transfer involving plasmids with mobilization gene cassettes containing antibiotic resistance genes [9, 10]. Due to this ability, K. pneumoniae is frequently pioneering acquisition of antibiotic resistance, putting this species in the front line of resistance against what at a given moment is considered a last resort drug, as in the cases of last generation cephalosporins, carbapenems or colistin [11, 12]. The possibility of tracking plasmids occurring in different bacterial hosts, detected in different patients or environmental niches, may have an important added value to unveil the major paths of antibiotic-resistance propagation
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