Abstract

Broad-spectrum beta-lactamase (BSBL)-producing Enterobacteriaceae impose public health threats. With increased popularity of zoos, exotic animals are brought in close proximity of humans, making them important BSBL reservoirs. However, not much is known on the presence of BSBLs in zoos in Western Europe. Fecal carriage of BSBL-producing Enterobacteriaceae was investigated in 38 zoo mammals from two Belgian zoos. Presence of bla-genes was investigated using PCR, followed by whole-genome sequencing and Fourier-transform infrared spectroscopy to cluster acquired resistance encoding genes and clonality of BSBL-producing isolates. Thirty-five putatively ceftiofur-resistant isolates were obtained from 52.6% of the zoo mammals. Most isolates were identified as E. coli (25/35), of which 64.0% showed multidrug resistance (MDR). Most frequently detected bla-genes were CTX-M-1 (17/25) and TEM-1 (4/25). Phylogenetic trees confirmed clustering of almost all E. coli isolates obtained from the same animal species. Clustering of five isolates from an Amur tiger, an Amur leopard, and a spectacled bear was observed in Zoo 1, as well as for five isolates from a spotted hyena and an African lion in Zoo 2. This might indicate clonal expansion of an E. coli strain in both zoos. In conclusion, MDR BSBL-producing bacteria were shown to be present in the fecal microbiota of zoo mammals in two zoos in Belgium. Further research is necessary to investigate if these bacteria pose zoonotic and health risks.

Highlights

  • The extensive use of β-lactams in human and veterinary medicine has favored global spread of broad-spectrum beta-lactamase (BSBL)-producing bacteria, especially in commensal Enterobacteriaceae [1]

  • Thirty-five putatively ceftiofur-resistant isolates were obtained from fecal samples of 20 zoo mammals, while fecal samples of the other 18 zoo mammals were culture-negative

  • Using MALDI-TOF MS, most of the isolates were identified as Enterobacteriaceae (80.0%, 28/35), namely E. coli (89.2%, 25/28), Escherichia marmotae (3.6%, 1/28), Klebsiella pneumoniae (3.6%, 1/28) and Citrobacter freundii (3.6%, 1/28)

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Summary

Introduction

The extensive use of β-lactams in human and veterinary medicine has favored global spread of broad-spectrum beta-lactamase (BSBL)-producing bacteria, especially in commensal Enterobacteriaceae [1]. These enzymes hydrolyze the amide bond of the β-lactam ring, rendering the antimicrobial ineffective [2]. Class A (TEM, SHV and CTX-M enzymes), C (AmpC enzymes) and D (OXA enzymes) function by the serine ester hydrolysis mechanism, while class B (metallo-β-lactamase (MBL) enzymes) use a zinc ion to attack the β-lactam ring. All. CTX-M-enzymes, most TEM- and SHV-enzymes and some OXA enzymes (i.e., OXA-10 and OXA-13 to OXA-19) are extended-spectrum β-lactamases (ESBLs).

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