# First Detection of International High-Risk blaKPC-2-Harbouring Escherichia coli Pandemic Lineage ST648 in Pet Food Packages

## TL;DR

This study reports the first detection of a dangerous antibiotic-resistant E. coli strain in pet food, raising concerns about its spread to pets and humans.

## Contribution

The first identification of blaKPC-2-harbouring E. coli ST648 in pet food, highlighting a new One Health risk.

## Key findings

- 15 multidrug-resistant CRE isolates were found in 86 pet food samples.
- Two E. coli isolates carried the blaKPC-2 gene and belonged to the high-risk ST648 lineage.
- The blaKPC-2 gene was located on an IncN plasmid within a Tn4401b transposon.

## Abstract

The continued worldwide increase in pet ownership has significantly boosted the growth of the pet food industry accompanied by new food safety risks and challenges. This study was designed to determine the occurrence and molecularly characterize multidrug-resistant (MDR) Enterobacterales in pet food. Eighty-six (86) packages of dry and wet pet food purchased in different retail stores were screened for carbapenem-resistant Enterobacterales (CRE). Antimicrobial susceptibility testing was performed by agar dilution technique using EUCAST/BrCAST recommendations. Blue-Carba test was further used to screen for carbapenemase-producing isolates. Isolated CRE strains were identified at the species level using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). Detection of carbapenemase-encoding genes was carried out by PCR, Sanger sequencing, and whole genome sequencing (WGS). A total of 15 (17.4%) MDR-CRE (Escherichia coli (n = 2), Enterobacter cloacae (n = 10), Leclercia adecarboxylata (n = 2), and Cronobacter spp. (n = 1)) were isolated from 86 pet food samples. In addition to being resistant to beta-lactams, the Gram-negative bacterial isolates were also resistant to aminoglycosides, fluoroquinolones, and tigecycline. Interestingly, two carbapenem-resistant E. coli isolates harboured blaKPC-2 gene. WGS analysis of the two blaKPC-2-producing E. coli isolates revealed that they both belong to ST648 and serotype O153:H2 group. The genetic context of the blaKPC-2 showed that they were carried by an IncN plasmid on a Tn4401b transposon element. To the best of our knowledge, this is the first description of blaKPC-2-harbouring E. coli ST648 pathogens in pet food. The detection of blaKPC-2-harbouring E. coli ST648 pandemic high-risk lineage in pet food is worrisome and a serious “One Health” issue. Therefore, pet food should be considered as a potential vehicle for the transmission of MDR pathogens to companion animals, and a risk factor for the dissemination of these bacterial pathogens to pet animals and their human guardians.