PMID- 38007076 OWN - NLM STAT- MEDLINE DCOM- 20240122 LR - 20240202 IS - 1096-0953 (Electronic) IS - 0013-9351 (Linking) VI - 242 DP - 2024 Feb 1 TI - Multidrug-resistant plasmid RP4 inhibits the nitrogen removal capacity of ammonia-oxidizing archaea, ammonia-oxidizing bacteria, and comammox in activated sludge. PG - 117739 LID - S0013-9351(23)02543-4 [pii] LID - 10.1016/j.envres.2023.117739 [doi] AB - In wastewater treatment plants (WWTPs), ammonia oxidation is primarily carried out by three types of ammonia oxidation microorganisms (AOMs): ammonia-oxidizing archaea (AOA), ammonia-oxidizing bacteria (AOB), and comammox (CMX). Antibiotic resistance genes (ARGs), which pose an important public health concern, have been identified at every stage of wastewater treatment. However, few studies have focused on the impact of ARGs on ammonia removal performance. Therefore, our study sought to investigate the effect of the representative multidrug-resistant plasmid RP4 on the functional microorganisms involved in ammonia oxidation. Using an inhibitor-based method, we first evaluated the contributions of AOA, AOB, and CMX to ammonia oxidation in activated sludge, which were determined to be 13.7%, 41.1%, and 39.1%, respectively. The inhibitory effects of C(2)H(2), C(8)H(14), and 3,4-dimethylpyrazole phosphate (DMPP) were then validated by qPCR. After adding donor strains to the sludge, fluorescence in situ hybridization (FISH) imaging analysis demonstrated the co-localization of RP4 plasmids and all three AOMs, thus confirming the horizontal gene transfer (HGT) of the RP4 plasmid among these microorganisms. Significant inhibitory effects of the RP4 plasmid on the ammonia nitrogen consumption of AOA, AOB, and CMX were also observed, with inhibition rates of 39.7%, 36.2%, and 49.7%, respectively. Moreover, amoA expression in AOB and CMX was variably inhibited by the RP4 plasmid, whereas AOA amoA expression was not inhibited. These results demonstrate the adverse environmental effects of the RP4 plasmid and provide indirect evidence supporting plasmid-mediated conjugation transfer from bacteria to archaea. CI - Copyright (c) 2023 Elsevier Inc. All rights reserved. FAU - Zhang, Zhaohui AU - Zhang Z AD - School of Environmental Science and Engineering, Tiangong University, State Key Laboratory of Separation Membranes and Membrane Processes, Binshui West Road 399, Xiqing District, Tianjin, 300387, China. Electronic address: zzh7448@126.com. FAU - Bo, Lin AU - Bo L AD - School of Environmental Science and Engineering, Tiangong University, State Key Laboratory of Separation Membranes and Membrane Processes, Binshui West Road 399, Xiqing District, Tianjin, 300387, China; Department of Hygienic Toxicology and Environmental Hygiene, Tianjin Institute of Environmental and Operational Medicine, Tianjin, 300050, China. FAU - Wang, Shang AU - Wang S AD - Department of Hygienic Toxicology and Environmental Hygiene, Tianjin Institute of Environmental and Operational Medicine, Tianjin, 300050, China. FAU - Li, Chenyu AU - Li C AD - Department of Hygienic Toxicology and Environmental Hygiene, Tianjin Institute of Environmental and Operational Medicine, Tianjin, 300050, China. FAU - Zhang, Xi AU - Zhang X AD - Department of Hygienic Toxicology and Environmental Hygiene, Tianjin Institute of Environmental and Operational Medicine, Tianjin, 300050, China. FAU - Xue, Bin AU - Xue B AD - Department of Hygienic Toxicology and Environmental Hygiene, Tianjin Institute of Environmental and Operational Medicine, Tianjin, 300050, China. FAU - Yang, Xiaobo AU - Yang X AD - Key Laboratory of Risk Assessment and Control for Environment & Food Safety, Tianjin, 300050, China. FAU - He, Xinxin AU - He X AD - Department of Hygienic Toxicology and Environmental Hygiene, Tianjin Institute of Environmental and Operational Medicine, Tianjin, 300050, China. FAU - Shen, Zhiqiang AU - Shen Z AD - Department of Hygienic Toxicology and Environmental Hygiene, Tianjin Institute of Environmental and Operational Medicine, Tianjin, 300050, China. FAU - Qiu, Zhigang AU - Qiu Z AD - Key Laboratory of Risk Assessment and Control for Environment & Food Safety, Tianjin, 300050, China. FAU - Zhao, Chen AU - Zhao C AD - Department of Hygienic Toxicology and Environmental Hygiene, Tianjin Institute of Environmental and Operational Medicine, Tianjin, 300050, China. Electronic address: zhaochen212@126.com. FAU - Wang, Jingfeng AU - Wang J AD - Key Laboratory of Risk Assessment and Control for Environment & Food Safety, Tianjin, 300050, China. Electronic address: jingfengwang@hotmail.com. LA - eng PT - Journal Article DEP - 20231123 PL - Netherlands TA - Environ Res JT - Environmental research JID - 0147621 RN - 0 (Sewage) RN - 7664-41-7 (Ammonia) RN - N762921K75 (Nitrogen) RN - 0 (Anti-Bacterial Agents) SB - IM MH - *Archaea/genetics/metabolism MH - Sewage/microbiology MH - Ammonia MH - Nitrogen/metabolism MH - Denitrification MH - In Situ Hybridization, Fluorescence MH - Oxidation-Reduction MH - Bacteria/genetics/metabolism MH - Plasmids/genetics MH - *Betaproteobacteria/genetics/metabolism MH - Anti-Bacterial Agents MH - Phylogeny MH - Soil Microbiology OTO - NOTNLM OT - AOA OT - AOB OT - Ammonia nitrogen consumption OT - Ammonia oxidation contribution OT - CMX OT - RP4 plasmid COIS- Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. EDAT- 2023/11/26 07:44 MHDA- 2024/01/22 06:43 CRDT- 2023/11/25 19:29 PHST- 2023/08/21 00:00 [received] PHST- 2023/11/09 00:00 [revised] PHST- 2023/11/17 00:00 [accepted] PHST- 2024/01/22 06:43 [medline] PHST- 2023/11/26 07:44 [pubmed] PHST- 2023/11/25 19:29 [entrez] AID - S0013-9351(23)02543-4 [pii] AID - 10.1016/j.envres.2023.117739 [doi] PST - ppublish SO - Environ Res. 2024 Feb 1;242:117739. doi: 10.1016/j.envres.2023.117739. Epub 2023 Nov 23.