{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T20:10:20Z","timestamp":1792613420906,"version":"4.5.0"},"reference-count":57,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T00:00:00Z","timestamp":1783382400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T00:00:00Z","timestamp":1783382400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2026,7,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Antimicrobial resistance monitoring in wastewater represents a key challenge, especially in light of the multiple methods available. Our study assessed the impact of selected sample processing techniques on microbial diversity and antimicrobial resistance gene recovery from wastewater. We analyzed 38 subsamples derived from 12 wastewater samples collected across three wastewater treatment plants over 4 months. We compared the effects of different centrifugation volumes (50, 250 mL), speeds (3,400 \u00d7\n                      <jats:italic toggle=\"yes\">g<\/jats:italic>\n                      vs 15,000 \u00d7\n                      <jats:italic toggle=\"yes\">g<\/jats:italic>\n                      ), and pellet versus filter concentration methods, as well as the performance of two DNA extraction kits\u2014DNeasy PowerSoil Pro Kit and DNeasy PowerLyzer Kit (Qiagen, DE)\u2014on wastewater samples. Additionally, we compared the performance of two DNA quantification instruments, NanoDrop One and Invitrogen Qubit 4 (Thermo Scientific, US). The analysis involved 16S rRNA gene sequencing on an Illumina MiSeq platform (PE250, 10M reads) and quantitative PCR (qPCR) of 11 selected antimicrobial resistance genes (ARGs). Higher centrifugation speeds and larger sample volumes lead to improved DNA yield. Pellets generally exhibited higher (1.32 times) ARG recoveries compared to filters. DNA quantification methods showed strong positive correlation (Pearson R = 0.78, Spearman \u03c1 = 0.81,\n                      <jats:italic toggle=\"yes\">P<\/jats:italic>\n                      &lt; 0.01) with each other and with qPCR data. The approach implemented in this study (centrifuging 200 mL samples at 15,000 \u00d7\n                      <jats:italic toggle=\"yes\">g<\/jats:italic>\n                      , followed by extraction with the DNeasy PowerSoil Pro Kit) shows promise for antimicrobial resistance (AMR) monitoring.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>To monitor the prevalence of antimicrobial resistance genes (ARGs) is crucial for anticipating and mitigating the spread of antimicrobial resistance (AMR). Urban wastewater provides community-level data to inform timely interventions. By establishing a method that balances efficiency and feasibility, this protocol represents a step toward a standardized wastewater surveillance (WS) framework for AMR, enhancing our ability to safeguard community health.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.02089-25","type":"journal-article","created":{"date-parts":[[2026,6,15]],"date-time":"2026-06-15T12:01:02Z","timestamp":1781524862000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Comparative analysis of wastewater sample processing methods for antimicrobial resistance surveillance"],"prefix":"10.1128","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2784-8007","authenticated-orcid":true,"given":"Helena Ferreira","family":"Leal","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0161xgx34","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology, Infectious Diseases and Immunology, Faculty of medicine, Universit\u00e9 de Montr\u00e9al (UdeM)","place":["Montr\u00e9al, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8144-9648","authenticated-orcid":false,"given":"Arthur","family":"Ouradou","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05f8d4e86","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Civil, Geological and Mining Engineering, Polytechnique Montr\u00e9al","place":["Montr\u00e9al, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nahla","family":"Koukene","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05f8d4e86","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Civil, Geological and Mining Engineering, Polytechnique Montr\u00e9al","place":["Montr\u00e9al, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amina","family":"Barka","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0161xgx34","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology, Infectious Diseases and Immunology, Faculty of medicine, Universit\u00e9 de Montr\u00e9al (UdeM)","place":["Montr\u00e9al, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1170-9475","authenticated-orcid":true,"given":"Caroline","family":"Quach","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0161xgx34","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology, Infectious Diseases and Immunology, Faculty of medicine, Universit\u00e9 de Montr\u00e9al (UdeM)","place":["Montr\u00e9al, Canada"]},{"id":[{"id":"https:\/\/ror.org\/03ej8cj80","id-type":"ROR","asserted-by":"publisher"}],"name":"Centre Hospitalier Universitaire Sainte-Justine (CHU Sainte-Justine)","place":["Montr\u00e9al, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1447-929X","authenticated-orcid":false,"given":"\u00c9milie","family":"B\u00e9dard","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05f8d4e86","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Civil, Geological and Mining Engineering, Polytechnique Montr\u00e9al","place":["Montr\u00e9al, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_5_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-34312-7"},{"key":"e_1_3_5_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-08853-3"},{"key":"e_1_3_5_4_2","doi-asserted-by":"publisher","DOI":"10.1097\/QCO.0000000000000929"},{"key":"e_1_3_5_5_2","doi-asserted-by":"publisher","unstructured":"N\u2019Guessan A Tsitouras A Sanchez-Quete F Goitom E Reiling SJ Galvez JH Nguyen TL Loan Nguyen HT Visentin F Hachad M et al.. 2022. Detection of prevalent SARS-CoV-2 variant lineages in wastewater and clinical sequences from cities in Qu\u00e9bec Canada. medRxiv. doi:10.1101\/2022.02.01.22270170","DOI":"10.1101\/2022.02.01.22270170"},{"key":"e_1_3_5_6_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0284370"},{"key":"e_1_3_5_7_2","doi-asserted-by":"publisher","DOI":"10.1101\/2023.10.30.23297765"},{"key":"e_1_3_5_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-41369-5"},{"key":"e_1_3_5_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.01853-22"},{"key":"e_1_3_5_10_2","doi-asserted-by":"publisher","unstructured":"Chau K Barker L Sims N Kasprzyk-Hordern B Budgell E Harriss E Crook D Read D Walker S Stoesser N. 2020. Wastewater surveillance of antimicrobial resistance in human populations: a systematic review. Preprints.org. doi:10.20944\/preprints202010.0267.v1","DOI":"10.20944\/preprints202010.0267.v1"},{"key":"e_1_3_5_11_2","doi-asserted-by":"publisher","DOI":"10.2166\/wh.2020.033"},{"key":"e_1_3_5_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2021.09.013"},{"key":"e_1_3_5_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.scitotenv.2023.162953"},{"key":"e_1_3_5_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.01428-23"},{"key":"e_1_3_5_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7012(03)00023-X"},{"key":"e_1_3_5_16_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0160523"},{"key":"e_1_3_5_17_2","doi-asserted-by":"publisher","DOI":"10.2166\/wh.2022.021"},{"key":"e_1_3_5_18_2","doi-asserted-by":"publisher","DOI":"10.3390\/ijerph20054555"},{"key":"e_1_3_5_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.watres.2019.05.102"},{"key":"e_1_3_5_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.scitotenv.2022.155576"},{"key":"e_1_3_5_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.06780-11"},{"key":"e_1_3_5_22_2","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.22-0427"},{"key":"e_1_3_5_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jenvman.2017.01.022"},{"key":"e_1_3_5_24_2","volume-title":"Cold Spring Harbor (NY)","author":"Russel D","year":"2001","unstructured":"Russel D, Sambrook J. 2001. Molecular cloning: a laboratory manual. In Cold Spring Harbor (NY), 3rd ed. Cold Spring Harbor Laboratory Press.","edition":"3"},{"key":"e_1_3_5_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jobb.2021.11.004"},{"key":"e_1_3_5_26_2","doi-asserted-by":"publisher","unstructured":"Faleye TOC Skidmore P Elyaderani A Adhikari S Kaiser N Smith A Yanez A Perleberg T Driver EM Halden RU Varsani A Scotch M. 2022. Impact of sample clarification by size exclusion on virus detection and diversity in wastewater-based epidemiology. medRxiv. doi:10.1101\/2022.09.25.22280344","DOI":"10.1101\/2022.09.25.22280344"},{"key":"e_1_3_5_27_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-08134-3"},{"key":"e_1_3_5_28_2","doi-asserted-by":"publisher","DOI":"10.1002\/agg2.20344"},{"key":"e_1_3_5_29_2","doi-asserted-by":"publisher","unstructured":"Haendiges J Timme R Ramachandran P Balkey M. 2020. DNA Quantification using the Qubit Fluorometer v1. Protocols.io. doi:10.17504\/protocols.io.bi8dkhs6","DOI":"10.17504\/protocols.io.bi8dkhs6"},{"key":"e_1_3_5_30_2","doi-asserted-by":"publisher","DOI":"10.3390\/analytica3030025"},{"key":"e_1_3_5_31_2","doi-asserted-by":"publisher","DOI":"10.3390\/medicina57121375"},{"key":"e_1_3_5_32_2","unstructured":"Scientific TF. 2024. Interpretation of nucleic acid 260\/280 ratios. Available from: https:\/\/assets.thermofisher.com\/TFS-Assets\/CAD\/Product-Bulletins\/T123-NanoDrop-Lite-Interpretation-of-Nucleic-Acid-260-280-Ratios.pdf. Retrieved 30 Oct 2024."},{"key":"e_1_3_5_33_2","volume-title":"Standard methods for the examination of water and wastewater","author":"Eaton AD","year":"2005","unstructured":"Eaton AD. 2005. Standard methods for the examination of water and wastewater. American Public Health Association."},{"key":"e_1_3_5_34_2","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.13023"},{"key":"e_1_3_5_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00592-14"},{"key":"e_1_3_5_36_2","unstructured":"Public Health Agency of Canada. 2024. 2024. Canadian Antimicrobial Resistance Surveillance System (CARSS): Interactive Database. Available from: https:\/\/health-infobase.canada.ca\/carss"},{"key":"e_1_3_5_37_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-25096-3"},{"key":"e_1_3_5_38_2","doi-asserted-by":"publisher","DOI":"10.1093\/femsec\/fiy130"},{"key":"e_1_3_5_39_2","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2020.00208"},{"key":"e_1_3_5_40_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2016.02137"},{"key":"e_1_3_5_41_2","doi-asserted-by":"publisher","DOI":"10.1093\/femsec\/fiw052"},{"key":"e_1_3_5_42_2","doi-asserted-by":"publisher","DOI":"10.1021\/es502615e"},{"key":"e_1_3_5_43_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2008.73"},{"key":"e_1_3_5_44_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks1219"},{"key":"e_1_3_5_45_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0061217"},{"key":"e_1_3_5_46_2","unstructured":"QIAGEN. 2020. DNeasy PowerLyzer PowerSoil Kit Handbook. Hilden Germany: QIAGEN. Available from: https:\/\/www.qiagen.com\/en-us\/resources\/download\/KitHandbook\/hb-2253-002-hb-dny-powerlyzer-powersoil-0120-ww"},{"key":"e_1_3_5_47_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-010-2770-5"},{"key":"e_1_3_5_48_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2013.05.008"},{"key":"e_1_3_5_49_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2017.07.007"},{"key":"e_1_3_5_50_2","doi-asserted-by":"publisher","DOI":"10.1186\/1472-6750-11-55"},{"key":"e_1_3_5_51_2","doi-asserted-by":"publisher","DOI":"10.1021\/es303685a"},{"key":"e_1_3_5_52_2","doi-asserted-by":"publisher","DOI":"10.1021\/es800720n"},{"key":"e_1_3_5_53_2","doi-asserted-by":"publisher","DOI":"10.2965\/jwet.18-086"},{"key":"e_1_3_5_54_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.scitotenv.2023.166442"},{"key":"e_1_3_5_55_2","doi-asserted-by":"publisher","unstructured":"Wu H-M. 2022. QIAGEN DNeasy PowerSoil Pro. doi:10.17504\/protocols.io.bp2l69411lqe\/v1","DOI":"10.17504\/protocols.io.bp2l69411lqe\/v1"},{"key":"e_1_3_5_56_2","doi-asserted-by":"publisher","DOI":"10.1111\/lam.12038"},{"key":"e_1_3_5_57_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02354-17"},{"key":"e_1_3_5_58_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0305650"}],"container-title":["Microbiology Spectrum"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.02089-25","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.02089-25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T01:02:52Z","timestamp":1788224572000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/spectrum.02089-25"}},"subtitle":[],"editor":[{"given":"Cezar M.","family":"Khursigara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,7,7]]},"references-count":57,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2026,7,7]]}},"alternative-id":["10.1128\/spectrum.02089-25"],"URL":"https:\/\/doi.org\/10.1128\/spectrum.02089-25","relation":{},"ISSN":["2165-0497"],"issn-type":[{"value":"2165-0497","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,7]]},"assertion":[{"value":"2025-11-27","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-04-14","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-06-15","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e02089-25"}}