Genomic landscape of blaGES-5- and blaGES-24-harboring Gram-negative bacteria from hospital wastewater: emergence of class 3 integron-associated blaGES-24 genes

Shino Takizawa; Eiji Soga; Wataru Hayashi; Kanae Sakaguchi; Shota Koide; Mizuki Tanabe; Tomohiro Denda; Yo Sugawara; Liansheng Yu; Shizuo Kayama; Motoyuki Sugai; Yukiko Nagano; Noriyuki Nagano

doi:10.1016/j.jgar.2022.09.005

Genomic landscape of bla_GES-5- and bla_GES-24-harboring Gram-negative bacteria from hospital wastewater: emergence of class 3 integron-associated bla_GES-24 genes

J Glob Antimicrob Resist. 2022 Dec:31:196-206. doi: 10.1016/j.jgar.2022.09.005. Epub 2022 Sep 28.

Authors

Affiliations

¹ Department of Health and Medical Sciences, Shinshu University Graduate School of Medicine, Matsumoto, Japan.
² Department of Health and Medical Sciences, Shinshu University Graduate School of Medicine, Matsumoto, Japan; Department of Laboratory Medicine, Nakatsugawa Municipal General Hospital, Nakatsugawa, Japan.
³ Department of Medical Sciences, Shinshu University Graduate School of Medicine, Science and Technology, Matsumoto, Japan.
⁴ Antimicrobial Resistance Research Center, National Institute of Infectious Diseases, Higashimurayama, Japan.
⁵ Department of Health and Medical Sciences, Shinshu University Graduate School of Medicine, Matsumoto, Japan; Department of Medical Sciences, Shinshu University Graduate School of Medicine, Science and Technology, Matsumoto, Japan. Electronic address: naganon@shinshu-u.ac.jp.

PMID: 36180037
DOI: 10.1016/j.jgar.2022.09.005

Abstract

Objectives: This study aimed to characterize Gram negative bacteria carrying bla_GES carbapenemase genes detected in wastewater from a hospital with no history of detection of clinical isolates producing GES carbapenemases.

Methods: Six hospital effluent samples were screened for carbapenemase-producing organisms (CPO) using CHROMagar mSuperCARBA and MacConkey agar with 1 µg/mL imipenem. Polymerase chain reaction (PCR) amplification and sequencing of carbapenemase genes, multilocus sequence typing, antimicrobial susceptibility testing, and whole-genome sequencing were performed.

Results: Among 21 CPO isolates, 11 Klebsiella spp. and 5 Enterobacter kobei isolates carried bla_GES-24, and 4 E. roggenkampii and 1 Pseudomonas aeruginosa isolates carried bla_GES-5. Genomic analysis of 8 representative isolates comprising 6 bla_GES-24-positive and 2 bla_GES-5-positive revealed that class 3 integrons with complete or defective Tn402-like transposition modules were predominantly associated with two tandem copies of bla_GES-24. Furthermore, a total of 5 new class 3 integrons, In3-18 to In3-22, were identified among 5 bla_GES-24 and 1 bla_GES-5 plasmids. One strain each of K. pneumoniae subsp. pneumoniae and K. quasipneumoniae subsp. similipneumoniae harboring bla_GES-24 plasmids also carried a rare bla_VEB-1-positive class 1 integron on a non-typeable plasmid, where these bla_VEB-1 plasmids had high sequence similarity. Virulence gene profiles differed between Klebsiella spp. and Enterobacter spp.; the former harbored type III fimbriae cluster, salmochelin, and T6SS type i2 gene clusters, while the latter had curli pili operon, aerobactin, T2SS gene clusters, and T6SS type i3 gene clusters.

Conclusion: Our findings confirmed the linkage of bla_GES-24 with rare Tn402-like class 3 integrons and the structural diversity of their gene cassette arrays.

Keywords: Enterobacter spp; Hospital wastewater; Klebsiella spp; bla(GES-24); bla(GES-5).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Genomics
Gram-Negative Bacteria
Hospitals
Integrons* / genetics
Wastewater*

Substances

Wastewater