Characterization of Acinetobacter baumannii from water and sludge line of secondary wastewater treatment plant.

The elimination of potentially pathogenic bacteria in wastewater treatment plants (WWTPs) attracts much attention in public health. Reports on the occurrence of the emerging hospital pathogen Acinetobacter baumannii in wastewaters do not include a continuous monitoring at all WWTP stages. The objective of this study was to characterize A. baumannii recovered from the water and sludge line of the secondary WWTP in Zagreb, Croatia over the period of one year. Recovery of A. baumannii was performed using CHROMagar Acinetobacter plates. Antimicrobial susceptibility testing was performed with broth microdilution and results interpreted using EUCAST breakpoints for clinical isolates of A. baumannii. Molecular characterization was performed by WGS and cgMLST. The secondary WWTP treating the urban wastewater is constantly receiving viable A. baumannii along with genes encoding carbapenem resistance, and emitting them via effluent into the environment. Furthermore, A. baumannii from influent are incorporated into activated sludge flocs in aeration basin. A. baumannii can survive the technological process of anaerobic mesophilic sludge digestion, and is finally destroyed in alkaline lime-treated stabilized sludge. The majority (102/119) of A. baumannii isolates were carbapenem-resistant, while antibiotic-susceptible isolates (17/119) were rarely recovered from all WWTP stages. Carbapenem-resistant isolates belonged to international clonal lineage IC2 carrying OXA-23 and IC1 carrying OXA-72, while the susceptible isolates belonged to IC5 or were unclustered. Increased resistance to antibiotics, together with the appearance of carbapenem- and even pandrug-resistant isolates in effluent as compared to influent wastewater, suggests the need of additional disinfection of effluent prior to its discharge into the natural recipient.