Discovering Novel Organophosphorus Compounds in Wastewater Treatment Plant Effluents through Suspect Screening and Nontarget Analysis.

Limited knowledge on the structure of emerging organophosphorus compounds (OPCs) hampers our comprehensive understanding of their environmental occurrence and potential risks. Through suspect and nontarget screening, combining data-dependent acquisition, data-independent acquisition, and parallel reaction monitoring modes, we identified 60 OPCs (17 traditional and 43 emerging compounds) in effluents of 14 wastewater treatment plants (WWTPs) in Beijing and Qinghai, China. These OPCs comprise 26 organophosphate triesters, 17 organophosphate diesters, 6 organophosphonates, 7 organothiophosphate esters, and 4 other OPCs. Notably, 14 suspect OPCs were newly identified in WWTP effluents, and 16 nontarget OPCs were newly discovered in environmental matrices. Specifically, the cyclic phosphonate, (5-ethyl-2-methyl-1,3,2-dioxaphosphorinan-5-yl)methyl dimethyl phosphonate P-oxide (PMMMPn), consistently appeared in all WWTP effluents, with semiquantitative concentrations ranging from 44.4 to 282 ng/L. Its analogue, di-PMMMPn, presented in 93% of wastewater samples. Compositional differences between the WWTP effluents of two cities were mainly attributed to emerging OPCs. Hazard and ecological risk assessment underscored the substantial contribution of chlorinated organophosphate esters and organothiophosphate esters to overall risks of OPCs in WWTP effluents. This study provides the most comprehensive OPC profiles in WWTP effluents to date, highlighting the need for further research on their occurrence, fate, and risks, particularly for chlorinated OPCs.

Suspect and Nontarget Screening Reveal the Underestimated Risks of Antibiotic Transformation Products in Wastewater Treatment Plant Effluents.

Antibiotics are anthropogenic contaminants with a global presence and of deep concern in aquatic environments, while less is known about the occurrence and risks of their transformation products (TPs). Herein, we developed a comprehensive suspect and nontarget screening workflow based on high-resolution mass spectrometry to identify unknown antibiotic TPs in wastewater treatment plant effluents. We identified 211 compounds (35 parent antibiotics and 176 TPs) at confidence levels of ≥3 and 107 TPs originated from macrolides. TPs were quantified by 17 TPs standards and semiquantified by the predicted response factors and accounted for 55.6-95.1% (76.7% on average) of the total concentrations of parents and TPs. 22.2%, 63.1%, and 18.8% of the identified TPs were estimated to be more persistent, mobile, and toxic than their parent antibiotics, respectively. Further ecological risk assessment based on concentrations and toxicity to aquatic organisms revealed that the cumulative risks of TPs were generally higher than those of parents. Despite the newly formed N-oxide TPs, the tertiary treatment process (mainly ozonation) could decrease the averaged 20.3% of concentrations and 36.2% of the risks of antibiotic-related compounds. This study highlights the necessity to include antibiotic TPs in environmental scrutiny and risk assessment of antibiotics in different aquatic environments.

Impacts of engineered nanoparticles and antibiotics on denitrification: Element cycling functional genes and antibiotic resistance genes.

The wide presence of antibiotics and minerals warrants their combined effects on the denitrification in natural aquatic environment. Herein, we investigated the effects of two antibiotics, sulfamethazine (SMZ) and chlortetracycline (CTC), on the reduction of NO3--N and accumulation of NO2--N in the absence and presence of engineered nanoparticles (NPs) (Al2O3, SiO2, and geothite) using 16 S rRNA sequencing and high-throughput quantitative PCR. The results showed that the addition of antibiotics inhibited the reduction of NO3--N by changing the bacterial community structure and reducing the abundance of denitrification genes, while engineered NPs promoted the denitrification by increasing the abundance of denitrification genes. In the binary systems, engineered NPs alleviated the inhibitory effect of antibiotics through enriching the denitrification genes and adsorbing antibiotics. Antibiotics and its combination with engineered NPs changed the composition of functional genes related to C, N, P, S metabolisms (p < 0.01). The addition of antibiotics and/or engineered NPs altered the bacterial community structure, which is dominated by the genera of Enterobacter (40.7-90.5%), Bacillus (4.9-58.5%), and Pseudomonas (0.21-12.7%). The significant relationship between denitrification, carbon metabolism genes, and antibiotic resistance genes revealed that the heterotrophic denitrifying bacteria may host the antibiotic resistance genes and denitrification genes simultaneously. The findings underscore the significance of engineered NPs in the toxicity assessment of pollutants, and provide a more realistic insight into the toxicity of antibiotics in the natural aquatic environment.

Black carbon and humic acid alleviate the toxicity of antibiotics to a cyanobacterium Synechocystis sp.

Natural organic matters (NOMs), omnipresent in natural water, challenge the toxicity assessment of pollutants to aquatic organisms due to their complex interactions with chemicals and organisms. Here, we investigated the combined toxicity of one solid NOM (black carbon, BC) or one soluble NOM (humic acid, HA) with antibiotics, roxithromycin (RTM) or gatifloxacin (GAT), to the cyanobacterium Synechocystis sp.. The NOMs alleviated the toxicity of RTM and GAT to Synechocystis sp., and BC had greater alleviation effects than HA due to its stronger adsorption to antibiotics. Antibiotics disturbed the photosynthesis of Synechocystis sp. significantly, which were also mitigated by BC and HA. Proteomic analysis showed that BC up-regulated the pathway of ribosome and photosynthetic antenna protein. GAT down-regulated the pathways of ABC transporter and oxidative phosphorylation. RTM interfered the pathway of porphyrin and chlorophyll metabolism. Furthermore, the addition of BC reduced the number of differentially expressed proteins caused by antibiotics, corroborating its mitigation effects on the toxicity of antibiotics. The disturbance of HA on the pathway of ABC transporters inhibited the internalization of RTM, thus decreasing its toxicity. This study underscores the significance of NOMs in mediating the toxicity of organic pollutants to aquatic organisms in natural waters.

Single and combined genotoxicity of metals and fluoroquinolones to zebrafish embryos at environmentally relevant concentrations.

Fluoroquinolones (FQs) are known to have genotoxicity to aquatic organisms. However, their genotoxicity mechanisms, individually and in combination with heavy metals, are poorly understood. Here, we investigated the single and joint genotoxicity of FQs, ciprofloxacin (CIP) and enrofloxacin (ENR), and metals (Cd and Cu) at environmentally relevant concentrations (0.2 µM) to zebrafish embryos. We found that FQs or/and metals induced genotoxicity (i.e., DNA damage and cell apoptosis) to zebrafish embryos. Compared with their single exposure, the combined exposure of FQs and metals elicited less ROS overproduction but higher genotoxicity, suggesting other toxicity mechanisms may also act in addition to oxidation stress. The upregulation of nucleic acid metabolites and the dysregulation of proteins confirmed the occurrence of DNA damage and apoptosis, and further revealed the inhibition of DNA repair by Cd and binding of DNA or DNA topoisomerase by FQs. This study deepens the knowledge on the responses of zebrafish embryos to exposure of multiple pollutants, and highlights the genotoxicity of FQs and heavy metals to aquatic organisms.

Antibiotic profiles and their relationships with multitrophic aquatic communities in an urban river.

Antibiotics have garnered worldwide attention due to their omnipresence and detrimental effects on aquatic organisms, yet their potential relationships with multitrophic aquatic communities in natural rivers remain largely unknown. Here, we examined 107 antibiotics in water and sediment from an urban river in Chengdu, Sichuan province (China). The bacterial, algal, macroinvertebrates, and fish communities were synchronously measured based on the environmental DNA (eDNA) metabarcoding approach, and their relationships with antibiotics were further investigated. The results showed that the total antibiotic concentrations ranged from 1.12 to 377 ng/L and from 7.95 to 145 ng/g in water and sediment, respectively. Significant seasonal variations in the concentrations and compositions of antibiotics in water were observed. eDNA metabarcoding revealed great compositional variations of bacterial, algal, macroinvertebrates, and fish communities along the river, and antibiotics had significant negative relationships with the community diversities of aquatic organisms (p < 0.05) except for fish. Meanwhile, significant negative correlations were observed between antibiotic concentrations and the relative abundances of essential metabolism pathways of bacteria, e.g., energy metabolism (p < 0.05), carbohydrate metabolism (p < 0.05), and lipid metabolism (p < 0.01). Moreover, antibiotics demonstrated greater effects on the function of bacterial community compared with environmental variables. The findings highlight the significance of eDNA metabarcoding approach in revealing the relationships between aquatic communities and antibiotics, and call for further studies on the effects of antibiotics on multitrophic aquatic communities in natural waters.

Integration of target, suspect, and nontarget screening with risk modeling for per- and polyfluoroalkyl substances prioritization in surface waters.

Though thousands of per- and polyfluoroalkyl substances (PFAS) have been on the global market, most research focused on only a small fraction, potentially resulting in underestimated environmental risks. Here, we used complementary target, suspect, and nontarget screening for quantifying and identifying the target and nontarget PFAS, respectively, and developed a risk model considering their specific properties to prioritize the PFAS in surface waters. Thirty-three PFAS were identified in surface water in the Chaobai river, Beijing. The suspect and nontarget screening by Orbitrap displayed a sensitivity of > 77%, indicating its good performance in identifying the PFAS in samples. We used triple quadrupole (QqQ) under multiple-reaction monitoring for quantifying PFAS with authentic standards due to its potentially high sensitivity. To quantify the nontarget PFAS without authentic standards, we trained a random forest regression model which presented the differences up to only 2.7 times between measured and predicted response factors (RFs). The maximum/minimum RF in each PFAS class was as high as 1.2-10.0 in Orbitrap and 1.7-22.3 in QqQ. A risk-based prioritization approach was developed to rank the identified PFAS, and four PFAS (i.e., perfluorooctanoic acid, hydrogenated perfluorohexanoic acid, bistriflimide, 6:2 fluorotelomer carboxylic acid) were flagged with high priority (risk index > 0.1) for remediation and management. Our study highlighted the importance of a quantification strategy during environmental scrutiny of PFAS, especially for nontarget PFAS without standards.

Decipher the molecular descriptors and mechanisms controlling sulfonamide adsorption onto mesoporous carbon: Density functional theory calculation and partial least-squares path modeling.

Mesoporous carbons (MCs) exhibit excellent removal efficiencies to various organic chemicals. However, how the properties of chemicals influence the adsorption mechanisms and further determine their adsorption onto MCs are poorly understood. We investigated the adsorption of 22 sulfonamides (SAs) onto four MCs, and further uncovered the major molecular descriptors and adsorption mechanisms influencing the adsorption by density functional theory (DFT) and partial least-squares path modeling (PLS-PM). The results revealed that the excess molar refraction (E), McGowan's molar volume (V), energy of the highest occupied molecular orbital (EHOMO), hardness (H), and most positive net charge on carbon atom (Qc+) were identified as the indirect factors affecting the distribution coefficient (logKD), by influencing the BE(π-π), BE(H), and logKow. BE(π-π) and logKow displayed significant direct impacts on logKD (p < 0.05), while BE(H) showed insignificant direct influences on logKD (p > 0.05). The PLS-PM results indicate the main driving forces for SAs adsorption including π-π interactions, hydrophobic effects, and hydrogen bonding. This study provides a new perspective on revealing the adsorption mechanisms, and the identified factors can be used to develop the quantitative model to further predict the adsorption of SAs onto MCs.

Antibiotic resistome and its driving factors in an urban river in northern China.

Urban rivers dynamically interfered by anthropogenic activities are considered as a vital reservoir of antibiotic resistance genes (ARGs). Here, a total of 198 ARGs and 12 mobile genetic elements (MGEs) were profiled in water and sediment from the Chaobai river, Beijing. The total abundances of ARGs (1.01 × 106-4.58 × 108 copies/L in water and 2.92 × 106-3.34 × 109 copies/g in sediment), which were dominated by beta-lactamase genes, exhibited significant seasonal variations (p < 0.05). Significant linear correlations between the total abundances of ARGs and MGEs were observed in both water and sediment (p < 0.01). Variance partitioning analysis disclosed that environmental variables (i.e., water temperature (WT), dissolved oxygen (DO), nutrients, metals, etc.) and antibiotics were the main contributors to the variations of ARGs and MGEs, and explained 55-80 % and 27-67 % of the total variations in ARGs and MGEs, respectively. The partial least-squares path model revealed the ARG abundances in water and sediment were affected by environmental variables and antibiotics both directly and indirectly but by MGEs directly. Moreover, random forest algorithm explored that WT, Ni, DO, Co, and polyether and macrolide antibiotics were the main drivers (>10 %) of ARGs dissemination in water, whereas the transposase genes of Tp614, tnpA, and IS613 were the main drivers of ARGs dissemination in both water and sediment. This study provides a comprehensive understanding of the driving factors for the ARGs dissemination in an urban river, which is of great significance for risk management of antibiotic resistome.

Occurrence, source apportionment, and pollution assessment of per- and polyfluoroalkyl substances in a river across rural and urban areas.

Forty-three novel and legacy per- and polyfluoroalkyl substances (PFASs) in water and sediments from the Chaobai River (Beijing) were quantified. The total PFASs concentrations varied from 0.04 to 31.3 ng/L in water with significant spatial but insignificant seasonal variations, and changed from 0.03 to 4.29 ng/g in sediment with insignificant spatial but significant seasonal variations. The PFASs concentrations in water from the upstream across the rural area reflected the background level due to the extremely low concentration and very few detected PFASs. The consumer products and metal plating/textile were the predominant pollution sources of PFASs in winter and summer, respectively, for both water and sediment samples. Integrating the determined baseline value, the distribution of PFASs concentrations, and the ecological risks of PFASs, three criteria were proposed, which divide the PFASs concentrations in water into four pollution levels, i.e., insignificant, low, medium, and high. According to the suggested criteria, 96.4% of the PFASs levels in upstream was insignificant pollution, which decreased to 50.4% in downstream and 50.8% in reservoirs. The PFASs in China's and world's surface waters demonstrated similar pollution patterns, with PFOA, PFOS, and PFHxA being the top 3 polluted PFASs. This study makes a small step forward the development of water quality standard for PFASs, which is of great importance for pollution control and risk management of PFASs.

Sediments alleviate the inhibition effects of antibiotics on denitrification: Functional gene, microbial community, and antibiotic resistance gene analysis.

Both antibiotics and sediments can affect the denitrification in aquatic systems. However, little is known how antibiotics influence the denitrification in the presence of sediments. Here, the effects of antibiotics (sulfamethoxazole, tetracycline and ofloxacin) on denitrification in the absence and presence of sediments were investigated. The influencing mechanisms were revealed by quantifying the denitrification functional genes (DNGs), 16S-seq of bacteria, and antibiotic resistance genes (ARGs). The results showed that the presence of antibiotics inhibited NO3-N reduction by decreasing the abundances of narG, nirK, nosZ, total DNGs, and denitrifying bacteria. However, the inhibition effect was alleviated by sediments, which promoted the growth of bacteria and decreased the selective pressure of antibiotics as the vector of bacteria and antibiotics, thus increasing the abundances of denitrifying bacteria and all the DNGs. Partial least-squares path model disclosed that antibiotics had negative effects on bacteria, ARGs and DNGs, while sediments had negative effects on ARGs but positive effects on bacteria and DNGs. The network analysis further revealed the close relation of the genera Bacillus, Acinetobacter, and Enterobacter with the ARGs and DNGs. The findings are helpful to understand the denitrification in antibiotic-polluted natural waters.