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While previous studies primarily focused on PAH concentrations, the impacts of PAH self-clustering have been largely overlooked, which will lead to inaccurate assessments of their ecological risks. This study evaluates the toxic effects of four prevalent PAH clusters on microbes with an emphasis on comparing the cluster sizes. Results revealed that over 95% of PAHs can form clusters in the aquatic environment, with smaller clusters more likely to form at lower concentrations and with fewer benzene rings. To quantify the toxic effects and understand underlying mechanisms, single-cell Raman-D2O was employed to link bacterial phenotypes with transcriptomic profiles. Bacteria exposed to smaller PAH clusters showed a 1%\u201310% reduction in metabolic activity, which was associated with a 1.8\u20132.9-fold increase in intracellular reactive oxygen species (ROS). Furthermore, when exposed to smaller PAH clusters, the expression of genes related to the ROS response and efflux pumps was upregulated by up to 6.33-fold and 4.97-fold, respectively, suggesting that smaller PAH clusters pose greater toxicity to microbes. These findings underscore the potentially overlooked risks of PAH clusters in environmental systems and deepen our understanding of the environmental fate and ecological risks of these contaminants.<\/jats:p>","DOI":"10.1021\/acs.est.4c10078","type":"journal-article","created":{"date-parts":[[2024,12,26]],"date-time":"2024-12-26T06:13:36Z","timestamp":1735193616000},"page":"224-233","source":"Crossref","is-referenced-by-count":17,"title":["Elevated Toxicity\nand High-Risk Impacts of Small Polycyclic\nAromatic Hydrocarbon Clusters on Microbes Compared to Large Clusters"],"prefix":"10.1021","volume":"59","author":[{"given":"Shuyue","family":"Zheng","sequence":"first","affiliation":[{"name":"Kunming University of Science & Technology , , ,","place":["Kunming, Yunnan, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jihong","family":"Dong","sequence":"additional","affiliation":[{"name":"Kunming University of Science & Technology , , ,","place":["Kunming, Yunnan, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2237-4711","authenticated-orcid":true,"given":"Quan","family":"Chen","sequence":"additional","affiliation":[{"name":"Kunming University of Science & Technology , , ,","place":["Kunming, Yunnan, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Wu","sequence":"additional","affiliation":[{"name":"Kunming University of Science & Technology , , ,","place":["Kunming, Yunnan, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0826-6423","authenticated-orcid":true,"given":"Dong","family":"Zhu","sequence":"additional","affiliation":[{"name":"Institute of Urban Environment, Chinese Academy of Sciences , , ,","place":["Xiamen, Fujian, China, 361021"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0708-8899","authenticated-orcid":true,"given":"Li","family":"Cui","sequence":"additional","affiliation":[{"name":"Institute of Urban Environment, Chinese Academy of Sciences , , ,","place":["Xiamen, Fujian, China, 361021"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philippe Frangois-Xavier","family":"Corvini","sequence":"additional","affiliation":[{"name":"Institute for Ecopreneurship, School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland , Gru\u0308ndenstrasse 40 , , ,","place":["Muttenz, Switzerland, 4132"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong-Zhe","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Urban Environment, Chinese Academy of Sciences , , ,","place":["Xiamen, Fujian, China, 361021"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3680-1451","authenticated-orcid":true,"given":"Bo","family":"Pan","sequence":"additional","affiliation":[{"name":"Kunming University of Science & Technology , , ,","place":["Kunming, Yunnan, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2024,12,26]]},"reference":[{"key":"2026081210104525800_cit1","doi-asserted-by":"publisher","first-page":"134937","DOI":"10.1016\/j.jhazmat.2024.134937","article-title":"PAHs contamination in ports: Status, sources and risks","volume":"475","author":"Tulcan","year":"2024","journal-title":"J. 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Technol."},{"issue":"2","key":"2026081210104525800_cit26","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1420406112","article-title":"Tracking heavy water (D2O) incorporation for identifying\nand sorting active microbial cells","volume":"112","author":"Berry","year":"2015","journal-title":"Proc. Natl.\nAcad. Sci. U. S. A."},{"issue":"18","key":"2026081210104525800_cit27","doi-asserted-by":"publisher","first-page":"11322","DOI":"10.1021\/acs.est.0c04292","article-title":"Temporal Dynamics of Antibiotic Resistome\nin the Plastisphere during Microbial Colonization","volume":"54","author":"Yang","year":"2020","journal-title":"Environ. Sci. 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Microbiol."},{"key":"2026081210104525800_cit47","doi-asserted-by":"publisher","first-page":"118407","DOI":"10.1016\/j.watres.2022.118407","article-title":"Response mechanisms\nof different antibiotic-resistant bacteria with different resistance\naction targets to the stress from photocatalytic oxidation","volume":"218","author":"Liu","year":"2022","journal-title":"Water Res."},{"issue":"1","key":"2026081210104525800_cit48","doi-asserted-by":"publisher","first-page":"3654","DOI":"10.1038\/s41467-024-48006-9","article-title":"Evolution of triclosan resistance modulates bacterial permissiveness\nto multidrug resistance plasmids and phages","volume":"15","author":"Yang","year":"2024","journal-title":"Nat. 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