Characterization of carbapenem resistance in environmental samples and Acinetobacter spp. isolates from wastewater and river water in Poland

Jakub Hubeny; Ewa Korzeniewska; Martyna Buta-Hubeny; Wiktor Zieliński; Damian Rolbiecki; Monika Harnisz

doi:10.1016/j.scitotenv.2022.153437

Characterization of carbapenem resistance in environmental samples and Acinetobacter spp. isolates from wastewater and river water in Poland

Sci Total Environ. 2022 May 20:822:153437. doi: 10.1016/j.scitotenv.2022.153437. Epub 2022 Feb 2.

Authors

Jakub Hubeny¹, Ewa Korzeniewska¹, Martyna Buta-Hubeny¹, Wiktor Zieliński¹, Damian Rolbiecki¹, Monika Harnisz²

Affiliations

¹ Department of Water Protection Engineering and Environmental Microbiology, Faculty of Geoengineering, University of Warmia and Mazury in Olsztyn, Prawocheńskiego 1, 10-720 Olsztyn, Poland.
² Department of Water Protection Engineering and Environmental Microbiology, Faculty of Geoengineering, University of Warmia and Mazury in Olsztyn, Prawocheńskiego 1, 10-720 Olsztyn, Poland. Electronic address: monika.harnisz@uwm.edu.pl.

PMID: 35122847
DOI: 10.1016/j.scitotenv.2022.153437

Abstract

The aim of this study was to analyze the prevalence of carbapenem resistance genes in Acinetobacter spp. isolated from wastewater in a municipal WWTP and to determine their spread from treated wastewater to river water with the use of conventional and molecular microbiology methods (qualitative and quantitative PCR and metagenomic analysis). Samples of untreated and treated wastewater and samples of river water obtained upstream and downstream from the wastewater discharge point were collected in 3 seasons (February, June, and September) of 2019. Acinetobacter spp. isolates were obtained by the culture method on the CHROMagar™ Acinetobacter medium. Additionally, environmental DNA was extracted from the samples for metagenomic and qPCR analyses. The presence of beta-lactam resistance genes (Ambler class B and D), insertion sequence ISAba1, and class I, II, and III integron-integrase genes was determined, and the bacterial taxonomic structure and wastewater and river samples was analyzed. Out of the 301 isolates obtained on the CHROMagar™ Acinetobacter medium, 258 belonged to the genus Acinetobacter, including 21 isolates that were identified as Acinetobacter baumannii. The highest number of Acinetobacter spp. and A. baumannii isolates were obtained from wastewater and river water samples collected in June and September. The ISAba1/bla_OXA-51 complex was identified in 13 isolates, which confirms the occurrence of carbapenem-resistance isolates in the analyzed samples. The number of Acinetobacter isolates carrying antibiotic resistance genes (ARGs) increased in river water samples collected downstream from the wastewater discharge point (48 out of 258 isolates - 18.6%) compared to river water samples collected upstream from the wastewater discharge point (34 out of 258 isolates - 13.2%), which suggests that WWTP is a source of pollution in the natural environment. The conducted research provides evidence that bacteria of the genus Acinetobacter may spread alarming beta-lactam resistance in the environment and, therefore, pose a serious epidemiological threat.

Keywords: Acinetobacter; Antibiotic resistance; Carbapenem resistance genes; Wastewater.

MeSH terms

Acinetobacter baumannii*
Anti-Bacterial Agents / analysis
Anti-Bacterial Agents / pharmacology
Bacterial Proteins / genetics
Carbapenems / pharmacology
Microbial Sensitivity Tests
Poland
Wastewater* / analysis
Water / analysis
beta-Lactamases / genetics

Substances

Anti-Bacterial Agents
Bacterial Proteins
Carbapenems
Waste Water
Water
beta-Lactamases