Imported One-Day-Old Chicks as Trojan Horses for Multidrug- Resistant Priority Pathogens Harboring mcr-9, rmtG, and Extended-Spectrum b-Lactamase Genes

Abstract

Antimicrobial resistance is a critical issue that is no longer restricted to hospital settings but also represents a growing problem involving intensive animal production systems. In this study, we performed a microbiological and molecular investigation of priority pathogens carrying transferable resistance genes to critical antimicrobials in 1-day-old chickens imported from Brazil to Uruguay. Bacterial identification was performed by matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry, and antibiotic susceptibility was determined by Sensititre. Antimicrobial resistance genes were sought by PCR, and clonality was assessed by pulsed-field gel electrophoresis (PFGE). Four multidrug-resistant (MDR) representative strains were sequenced by an Illumina and/or Oxford Nanopore Technologies device. Twenty-eight MDR isolates were identified as Escherichia coli (n = 14), Enterobacter cloacae (n = 11), or Klebsiella pneumoniae (n = 3). While resistance to oxyiminocephalosporins was due to blaCTX-M-2, blaCTX-M-8, blaCTX-M-15, blaCTX-M-55, and blaCMY-2, plasmid-mediated quinolone resistance was associated with the qnrB19, qnrE1, and qnrB2 genes. Finally, resistance to aminoglycosides and fosfomycin was due to the presence of 16S rRNA methyltransferase rmtG and fosA-type genes, respectively. Short- and long-read genome sequencing of E. cloacae strain ODC_Eclo3 revealed the presence of IncQ/rmtG (pUR-EC3.1; 7,400 bp), IncHI2A/mcr- 9.1/blaCTX-M-2 (pUR-EC3.2, ST16 [pMLST; 408,436 bp), and IncN2/qnrB19/aacC3/aph(30)- Ib (pUR-EC3.3) resistance plasmids. Strikingly, the blaCTX-M-2 gene was carried by a novel Tn1696-like composite transposon designated Tn7337. In summary, we report that imported 1-day-old chicks can act as Trojan horses for the hidden spread of WHO critical-priority MDR pathogens harboring mcr-9, rmtG, and extended-spectrum b-lactamase genes in poultry farms, which is a critical issue from a One Health perspective.