Print Email Facebook Twitter Metagenomic profiling and transfer dynamics of antibiotic resistance determinants in a full-scale granular sludge wastewater treatment plant Title Metagenomic profiling and transfer dynamics of antibiotic resistance determinants in a full-scale granular sludge wastewater treatment plant Author Calderon Franco, D. (TU Delft BT/Environmental Biotechnology) Sarelse, R.G. (Student TU Delft) Christou, S. (Student TU Delft) Pronk, M. (TU Delft BT/Environmental Biotechnology) van Loosdrecht, Mark C.M. (TU Delft BT/Environmental Biotechnology) Abeel, T.E.P.M.F. (TU Delft Pattern Recognition and Bioinformatics) Weissbrodt, D.G. (TU Delft BT/Environmental Biotechnology) Date 2022 Abstract In the One Health context, wastewater treatment plants (WWTPs) are central to safeguarding water resources. Nonetheless, many questions remain about their effectiveness in preventing antimicrobial resistance (AMR) dissemination. Most surveillance studies monitor the levels and removal of selected antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) in intracellular DNA (iDNA) extracted from WWTP influents and effluents. The role of extracellular free DNA (exDNA) in wastewater is mostly overlooked. This study analyzed the transfer of ARGs and MGEs in a full-scale Nereda® reactor removing nutrients with aerobic granular sludge. We tracked the composition and fate of the iDNA and exDNA pools of influent, sludge, and effluent samples. Metagenomics was used to profile the microbiome, resistome, and mobilome signatures of iDNA and exDNA extracts. Selected ARGs and MGEs were analyzed by qPCR. From 2,840 ARGs identified, the genes arr-3 (2%), tetC (1.6%), sul1 (1.5%), oqxB (1.2%), and aph(3")-Ib (1.2%) were the most abundant among all sampling points and bioaggregates. Pseudomonas, Acinetobacter, Aeromonas, Acidovorax, Rhodoferax, and Streptomyces populations were the main potential hosts of ARGs in the sludge. In the effluent, 478 resistance determinants were detected, of which 89% were from exDNA potentially released by cell lysis during aeration in the reactor. MGEs and multiple ARGs were co-localized on the same extracellular genetic contigs. Total intracellular ARGs decreased 3-42% due to wastewater treatment. However, the ermB and sul1 genes increased by 2 and 1 log gene copies mL−1, respectively, in exDNA from influent to effluent. The exDNA fractions need to be considered in AMR surveillance, risk assessment, and mitigation strategies. Subject Antibiotic resistanceMetagenomicswastewatermobile genetic elementsquantitative PCR To reference this document use: http://resolver.tudelft.nl/uuid:e859be6b-f1d5-40e5-a601-552f8968bce8 DOI https://doi.org/10.1016/j.watres.2022.118571 ISSN 0043-1354 Source Water Research, 219 Part of collection Institutional Repository Document type journal article Rights © 2022 D. Calderon Franco, R.G. Sarelse, S. Christou, M. Pronk, Mark C.M. van Loosdrecht, T.E.P.M.F. Abeel, D.G. Weissbrodt Files PDF 1_s2.0_S0043135422005243_main.pdf 2.52 MB Close viewer /islandora/object/uuid:e859be6b-f1d5-40e5-a601-552f8968bce8/datastream/OBJ/view