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Nanoparticle (NP) assessment in sludge materials, although of growing importance in eco- and biotoxicity studies, is commonly overlooked and, at best, understudied. In the present study, sewage sludge samples from across the mega-city of Shanghai, China were investigated for the first time using a sequential extraction method coupled with single particle inductively coupled plasma mass spectrometry (SP-ICP-MS) to quantify the abundance of metal-containing NPs in the extraction fractions and transmission electron microscopy to specifically identify the nanophases present. In general, most sludges observed showed high concentrations of Cr, Cu, Cd, Ni, Zn, and Pb, exceeding the maximum permitted values in the national application standard of acid soil in China. NPs in these sludges contribute little to the volume and mass but account for about half of the total particle number. Based on electron microscopy techniques, various NPs were further identified, including Ti-, Fe-, Zn-, Sn-, and Pb-containing NPs. All NPs, ignored by traditional metal risk evaluation methods, were observed at a concentration of 10 -10 particles/g within the bioavailable fraction of metals. These results indicate the underestimate or misestimation in evaluating the environmental risks of metals based on traditional sequential extraction methods. A new approach for the environmental risk assessment of metals, including NPs, is urgently needed.

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SummaryMembers of the marine bacterial genus Pseudoalteromonas are efficient producers of antifouling agents that exert inhibitory effects on the settlement of invertebrate larvae. The production of pigmented secondary metabolites by Pseudoalteromonas has been suggested to play a role in surface colonization. However, the physiological characteristics of the pigments produced by Pseudoalteromonas remain largely unknown. In this study, we identified and characterized a genetic variant that hyperproduces a dark‐brown pigment and was generated during Pseudoalteromonas lipolytica biofilm formation. Through whole‐genome resequencing combined with targeted gene deletion and complementation, we found that a point mutation within the hmgA gene, which encodes homogentisate 1,2‐dioxygenase, is solely responsible for the overproduction of the dark‐brown pigment pyomelanin. In P. lipolytica, inactivation of the hmgA gene led to the formation of extracellular pyomelanin and greatly reduced larval settlement and metamorphosis of the mussel Mytilus coruscus. Additionally, the extracted pyomelanin from the hmgA deletion mutant and the in vitro‐synthesized pyomelanin also reduced larval settlement and metamorphosis of M. coruscus, suggesting that extracellular pyomelanin released from marine Pseudoalteromonas biofilm can inhibit the settlement of fouling organisms.

The abundances of ARGs increased after exposure to ZnO NPs, especially at low doses (0.2 and 1 mg L−1). Compared to MGEs and dissolved Zn, the microbial community contributed more to the variations in ARGs in estuarine waters when exposed to ZnO NPs.

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