Detection of IncN‐pST15 one‐health plasmid harbouring bla KPC‐2 in a hypermucoviscous Klebsiella pneumoniae CG258 isolated from an infected dog, Brazil

Abstract The emergence and rapid spread of carbapenemase‐producing Enterobacterales represents a serious public health concern. Critically, these global priority bacteria have begun to be reported in companion animals, implying a potential risk of cross‐transmission between humans and pets. Using long‐read (MinION) and short‐read (Illumina) sequencing technologies, we have identified and characterized a hypermucoviscous KPC‐2‐producing Klebsiella pneumoniae strain belonging to the high‐risk international clone ST11/CG258, in a dog with urinary tract infection. Strikingly, the bla KPC‐2 gene was carried by a 54‐kb IncN plasmid assignated to ST15, which shared 99.8 and 96.8% pairwise identity with IncN‐pST15 plasmids from human and environmental K. pneumoniae strains, respectively; all come from an area with high endemicity of KPC‐2. Our findings suggest that IncN‐pST15 plasmids conferring carbapenem resistance can play as important a role as clonal transmission of K. pneumoniae, representing another major challenge for One Health.


| INTRODUC TI ON
Epidemiological studies have revealed that carbapenemaseproducing Enterobacterales have emerged in healthy and sick animals, and community settings (Kelly et al., 2017;Wang et al., 2020;Zhang et al., 2019), implying a potential risk of transmission of these pathogens between humans and companion animals (Grönthal et al., 2018;Sellera & Lincopan, 2019). Additionally, the transfer of carbapenems resistance genes can be facilitated by mobile genetic elements (e.g. plasmids and transposons), which is a concerning possibility (Baquero et al., 2019;Brandt et al., 2019).
KPC family has been the most widespread of all carbapenemases associated with Enterobacterales (van Duin & Doi, 2017). The occurrence of KPC-producing bacteria in human hospital settings has rendered nosocomial infections particularly difficult to treat or even untreatable (Wang et al., 2016). To date, the identification of KPC producers in companion animals has been sporadically reported from dogs in Brazil (KPC-2-producing Escherichia coli) and United States (KPC-4-producing Enterobacter xiangfangensis) Sellera et al., 2018).
In this study, under a 'One Health' view, we report the identification of a KPC-2-positive Klebsiella pneumoniae belonging to the international high-risk clone sequence type 11/clonal group 258 (ST11/ CG258) in a dog suffering from urinary tract infection, highlighting that IncN-pST15 plasmids carrying bla KPC-2 genes are spreading among human, animal and environmental clonally unrelated K. pneumoniae strains.

| MATERIAL S AND ME THODS
In 2019, during a Brazilian surveillance study (OneBR project), conducted to characterize the burden of antimicrobial resistance associated with critical WHO priority pathogens, a carbapenemresistant K. pneumoniae strain (PVT01) identified by BD Phoenix (BD Diagnostics, Sparks, MD, USA) was isolated from a urine culture of a 9-year-old female Spitz dog suffering from urinary tract infection.
In summary, to the best of our knowledge, this is the first report of KPC-positive K. pneumoniae ST11/CG258 isolated from a pet. The emergence of KPC-2-producing bacteria in companion animals is an important public health issue that denotes that pets are a neglected reservoir for critical priority pathogens in the community, and susceptible hosts for acquisition of untreatable or difficult-to-treat infections (Abraham et al., 2014;Köck et al., 2018;Pomba et al., 2017;Sellera & Lincopan, 2019). In this regard, IncN-pST15 plasmids conferring carbapenem resistance can play as important a role as clonal transmission of K. pneumoniae, representing another major challenge for One Health. Therefore, surveillance studies should investigate similarities of plasmids circulating at the human-environmentanimal interface in addition to clonal transmission. Brazil) and CEFAP-GENIAL facility for kindly supplying antibiotic discs for susceptibility testing and Illumina sequencing, respectively.

CO N FLI C T O F I NTE R E S T
The authors have no conflict of interest to declare.

E TH I C A L A PPROVA L
The authors confirm that the ethical policies of the journal, as noted on the journal's author guidelines page, have been adhered to. No ethical approval was required for this specific study.

DATA AVA I L A B I L I T Y S TAT E M E N T
All data generated or used during the study appear in the submitted article. The data that support the findings of this study are avail- Nilton Lincopan https://orcid.org/0000-0003-0161-5800