Antibiotic resistance genes, antibiotic residues, and microplastics in influent and effluent wastewater from treatment plants in Norway, Iceland, and Finland

Monitoring antimicrobial resistance genes (ARGs) in wastewater influents (pre-treatment) and effluents (post-treatment) provides insights into community-level circulation, potential amplification during treatment, and risks associated with gene release into surface waters. Pollutants such as antibiotic residues and microplastics (MPs) may influence ARG dynamics, highlighting the need to assess their dynamics across wastewater environments. In this study, we analyzed ARGs and bacterial communities using Oxford Nanopore (ONP) metagenomics and qPCR in wastewater samples from Mekjarvik (Norway), Reykjavik (Iceland), and Mariehamn (Åland, Finland). Antibiotic residues were quantified via High-Performance Liquid Chromatography (HPLC), and MPs were characterized using Micro-Fourier Transform Infrared Spectroscopy (μ-FTIR) in Mekjarvik and Reykjavik. Metagenomic analysis identified 193 unique ARGs, with the highest average (±SD) in Reykjavik (66.3 ± 4.1), followed by Mekjarvik (61.3 ± 14.1) and Mariehamn (18.0 ± 2.2). ONP sequencing revealed that many ARGs were plasmid-associated, co-occurring with metal stress genes. Common plasmids were Col440I , IncQ2 , and ColRNAI . Mercury-related genes dominated metal stress genes (64.9%), followed by multimetal (23.7%) and copper (6.4%). Of 45 antibiotics screened, only sulfamethoxazole and sulfapyridine were consistently detected. Polyethylene (∼60%) was the dominant MP type; Reykjavik influent had the highest MP load (8200 MPs/m 3 ). While treatment reduced ARGs, antibiotic residues, and larger MPs, it was less effective against fine particles and key ARGs, including carbapenemase- and ESBL-associated genes. Clinically relevant ARGs and potential pathogens (e.g., Acinetobacter baumannii , Pseudomonas aeruginosa ) persisted in effluents, highlighting risks to downstream ecosystems. These findings underscore the need for regular monitoring of both influents and effluents to assess treatment performance and safeguard environmental health.