Complex Environmental Samples Research Articles

Aptamer based hybrid monolithic pipette tips supported by melamine sponge for enrichment of proteins

BackgroundThe convenient preparation and application of functionalized organic-inorganic hybrid monolithic materials have obtained substantial interest in the pretreatment of complex samples by solid-phase extraction (SPE). Compared to the in-tube solid-phase microextraction in fused-silica capillaries, micro SPE in plastic pipette tips have fascinating merits for the easily operated enrichment of trace target analytes from biological samples. However, the poor compatibility of organic-inorganic hybrid monoliths with plastics leads to the rare appearance of commercial hybrid monolithic pipette tips (HMPTs). Therefore, how to synthesize the organic-inorganic hybrid monolithic materials with better extraction performance in plastic pipette tips becomes a challenge. ResultsWe develop a facile and cheap strategy to immobilize organic-inorganic hybrid monoliths in pipette tips. Melamine sponge was employed as the supporting skeleton to in situ assemble amine- and thiol-bifunctionalized hybrid monolithic material via “one pot” in a pipette tip, and gold nanoparticles (GNPs) and thiol-modified aptamer against human α-thrombin were sequentially attached to the hybrid monolith within the HMPTs. The average coverage density of the aptamer with GNPs as an intermediary reached as high as 818.5 pmol μL−1. The enriched thrombin concentration was determined by a sensitive enzymatic chromogenic assay with the limit of detection of 2 nM. The extraction recovery of thrombin at 10 nM in human serum was 86.1 % with a relative standard deviation of 6.1 %. This proposed protocol has been applied to the enrichment and determination of thrombin in real serum sample with strong anti-interference ability, low limit of detection and high recovery. SignificanceThe amine- and thiol-bifunctionalized HMPTs prepared with sponge as the skeleton frame provided a novel substrate material to decorate aptamers for efficient enrichment of proteins. This enlightens us that we can take advantage of the tunability of sponge assisted HMPTs to produce and tailor a variety of micro SPE pipette tips for broader applications on the analysis of trace targets in complex biological, clinic and environmental samples.

Novel biocompatible N-rich AIE fluorescent probe for live cell imaging and visual onsite detection of uranium.

A sensitive and biocompatible N-rich probe for rapid visualuranium detectionwas constructed by grafting two trianiline groups to 2,6-bis(aminomethyl)pyridine. Possessing excellent aggregation-induced emission (AIE) property and the advantages to form multidentate chelate with U selectively, the probe has been applied successfully to visualize uranium in complex environmental water samples and living cells, demonstrating outstanding anti-interference ability against large equivalent of different ions over a wide effective pH range. A large linear range (1.0 × 10-7-9.0 × 10-7mol/L) and low detection limit (72.6nmol/L, 17.28ppb) were achieved for the visual determination of uranium. The recognition mechanism, photophysical properties, analytical performance and cytotoxicity were systematically investigated, demonstrating highpotential for fast risk assessment of uranium pollution in field and in vivo.

Microplastic analysis in sediments of the Elbe River by electrostatic separation and differential scanning calorimetry

This study presents the most extensive investigation of microplastic (MP) contents in sediment from the Elbe River. We employed electrostatic separation (ES) and differential scanning calorimetry (DSC) to overcome limitations of sample throughput and time-consuming analysis. In total 43 sediment samples were collected using a Van-Veen grab. Subsequently, coarse materials (d10 > 100 μm) and fine materials (d10 ≤ 100 μm) were enriched using ES and density separation. DSC was utilized for MP identification and quantification, based on the phase-transition signals of eight different polymers. MP presence was detected in 25 samples, with successful quantification in 12 samples. The MP content in coarse material samples from shoreline areas ranged from 0.52 to 1.30 mg/kg, while in fine material samples from harbor basins, it ranged from 5.0 to 44.6 mg/kg. The most prevalent polymers identified were LD-PE, HD-PE, PP, and PCL. These findings confirmed the suitability of DSC for analyzing MP in complex environmental samples. MP hotspots were identified in harbor basins, where natural sedimentation processes and increased anthropogenic activities contribute to MP accumulation. Additionally, industrial sewage potentially contributed to MP content in sediment samples. The highest pollution levels were observed in the middle Elbe, between the confluences of Mulde and Havel. Lowest MP contents were found in the lower Elbe, potentially influenced by tides. Future studies should focus on holistic investigations of selected river sections, encompassing sediment, water, and biota samples, rather than the entire catchment area. This approach would facilitate the generation of spatiotemporal data on MP distribution in freshwater streams. In addition, more research is needed to explore potential interactions between different MP and sediment types during DSC measurements.

Silver Nanoparticles In Situ Enhanced Electrochemiluminescence of the Porphyrin Organic Matrix for Highly Sensitive and Rapid Monitoring of Tetracycline Residues.

Accurate monitoring of tetracycline (TC) residues in the environment is crucial for avoiding contaminant risk. Herein, a novel TC biosensor was facilely designed by integrating silver nanoparticles (Ag NPs) into the porphyrin metal-organic matrix (Ag@AgPOM) as a bifunctional electrochemiluminescence (ECL) probe. Different from the step-by-step synthesis of the co-reaction accelerator and ECL emitter, the co-reaction accelerators Ag NPs were in situ-grown on the surface of 5,10,15,20-tetrakis (4-carboxyphenyl) porphyrin (TCPP) via a simple one-pot approach. Symbiotic Ag NPs on Ag@AgPOM formed an intimate interface and increased the collision efficiency of the ECL reaction, achieving the ECL enhancement of TCPP. Under the optimized conditions, the ternary ECL biosensor showed a wide linear detection range toward TC with a low detection limit of 0.14 fmol L-1. Compared with the traditional HPLC and ELISA methods, satisfied analytical adaptability made this sensing strategy feasible to monitor TC in complex environmental samples.

Aptasensors for the Detection of Environmental Contaminants of High Concern in Water Bodies: A Systematic Review

With the advancement of technology and increasing industrial activity, anthropogenic contaminants are currently detected where there is no record of their presence or insufficient information about their toxicological impact. Consequently, there are not sufficiently robust local or global regulations, the ecotoxicological and human health risks are critical, and they may not be routinely monitored despite being ubiquitous. The interest in studying environmental contaminants, including micropollutants and emerging contaminants, in complex environmental water samples has grown in the last decade. Due to the concentrations in which they are typically found in the environment and the rapid global dispersion, the detection procedures for these substances must be capable of measuring very low concentrations. Many efforts have been made to improve remediation procedures or develop novel analytical methods for their determination. Although there are several robust and reliable standard analytical techniques for their monitoring, pollutant contamination requires simple and inexpensive methods for massive, in situ monitoring campaigns. In this regard, biosensors have emerged as devices with high selectivity, sensitivity, easy operation, and short analysis times. Aptasensors are biosensors based on a nucleic acid recognition element (aptamer). Due to their synthetic nature, stability, and easy production, aptamers are frequently employed to develop bioassays. This work presents a systematic review of the trends in using aptasensors for detecting environmental contaminants present in environmental water samples, as well as the estimation of the potential technological contribution these devices might give to environmental monitoring.

Approaches for attaining clean bacterial fractions from complex environmental samples

Marine bacteria have been targeted by industry and pharmaceutics as genetic resources for highly active enzymes or novel lead compounds. Although numerous techniques have been introduced to isolate useful bacteria from the environment, we are still highly dependent on the conventional direct cultivation method to attain pure cultures. However, efficient bacterial isolation is hindered by several factors, including the presence of impurities. In this work, to demonstrate the significance of removing impurities and their impact on bacterial isolation, we employed two approaches: dielectrophoresis (DEP) and fluorescent D-amino acids (FDAA). We successfully attained clean bacterial fractions applicable for downstream processing using these approaches, uniquely designed to identify bacteria based on their characteristics and features. The diversity of bacteria attained by both approaches was investigated using 16S rRNA sequencing and compared to that attained by the standard differential centrifugation method. In addition, the viability of the isolates was also determined via direct cultivation. As a result, the separation of bacteria from impurities allowed for the identification of novel and useful bacteria unique to each approach. Successful cultivation also suggested that both approaches were applicable for attaining viable bacteria. In conclusion, removing impurities to attain clean bacterial fractions promotes the isolation of novel bacteria and thus could aid in the successful isolation of useful bacteria within complex environmental samples.

Sacrificial template synthesis of hollow sulfonate group functionalized microporous organic network for efficient solid phase extraction of sulfonamide antibiotics from milk and honey samples

The highly conjugated and hydrophobic characteristics of microporous organic networks (MONs) have largely impeded their broad applications in sample pretreatment especially for the polar or ionic analytes. In this work, a novel uniform hollow shaped sulfonate group functionalized MON (H-MON-SO3H-2) was synthesized via the sacrificial template method for the efficient solid phase extraction (SPE) of sulfonamides (SAs) from environmental water, milk, and honey samples prior to HPLC analysis. H-MON-SO3H-2 exhibited large specific surface area, penetrable space, good stability, and numerous hydrogen bonding, electrostatic, hydrophobic and π-π interaction sites, allowing sensitive SPE of SAs with wide linear range (0.150-1000 μg L−1), low limit of detection (0.045-0.188 μg L−1), good precisions (intra-day and inter-day RSD < 7.3%, n = 5), large enrichment factors (95.7-98.5), high adsorption capacities (250.4-545.0 mg g−1), and satisfactory reusability (more than 80 times). Moreover, the established method was successfully applied to extract SAs from spiked samples with the recoveries of 86.1-104.3%. This work demonstrated the great potential of H-MON-SO3H-2 in the efficient SPE of trace SAs in complex environmental water and food samples and revealed the prospect of hollow MONs in sample pretreatment.

Research progress of novel functional materials in extraction of algal toxins

Algal toxins are secondary metabolites produced by harmful algae; these metabolites are characterized with strong toxicity, diverse structure and bioaccumulation. Aquatic organisms that feed on harmful algae can accumulate algal toxins in their bodies, and the consumption of these organisms by humans can cause symptoms of paralysis, diarrhea, and even death. The onset of poisoning can occur within as little as 30 min; in many cases, no suitable antidote for algal toxins is available. Thus, algal toxins present significant threats to human health, the aquaculture industry, and aquatic ecosystems. Because the potential risks of algal toxins are a critical issue, these toxins have become a research hotspot. The water environment and various types of aquatic products should be monitored and analyzed to ensure their safety. However, because of possible matrix effects and the low content of algal toxins in actual samples, an efficient pretreatment method is necessary prior to instrumental analyses. Efficient sample pretreatment techniques can not only reduce or eliminate interferences from the sample matrix during analysis but also enrich the target analytes to meet the detection limit of the analytical instrument, thereby ensuring the sensitivity and accuracy of the detection method. In recent years, sample pretreatment techniques such as solid-phase extraction (SPE), solid-phase microextraction (SPME), magnetic SPE (MSPE), dispersive SPE (DSPE), and pipette tip-based SPE (PT-SPE) have gained wide attention in the field of algal-toxin separation and analysis. The performance of these pretreatment techniques largely depends on the characteristics of the extraction materials. Given the diverse physicochemical properties of algal toxins, including their different molecular sizes, hydrophobicity/hydrophilicity, and charges, the design and preparation of materials suitable for algal-toxin extraction is an essential undertaking. The optimal extraction material should be capable of reversible algal-toxin adsorption and preferably possess a porous structure with a large surface area to allow for high recovery rates and good interfacial contact with the toxins. Additionally, the extraction material should exhibit good chemical stability in the sample solution and elution solvent within the working pH range; otherwise, it may dissolve or lose its functional groups. Many research efforts have sought to develop novel adsorbent materials with these properties in the separation and analysis of algal toxins, focusing on carbon-based materials, metal organic frameworks (MOFs), covalent organic frameworks (COFs), molecularly imprinted polymers (MIPs), and their functionalized counterparts. Carbon-based materials, MOFs, and COFs have advantages such as large surface areas and abundant adsorption sites. These extraction materials are widely used in the separation and analysis of target substances in complex environmental, biological, and food samples owing to their excellent performance and unique microstructure. They are also the main adsorbents used for the extraction of algal toxins. These extraction materials play an essential role in the extraction of algal toxins, but they also present a number of limitations: (1) Carbon-based materials, MOFs, and COFs have relatively poor selective-adsorption ability towards target substances; (2) Most MOFs are unstable in aqueous solutions and challenging to apply during extraction from water-based sample solutions; (3) COFs mainly consist of lightweight elements, rendering them difficult to completely separate from sample solutions using centrifugal force, which limits their application range; (4) Although MIPs have good selectivity, issues such as template-molecule loss, slow mass-transfer rates, and low adsorption capacity must be addressed. Therefore, the design and preparation of novel functionalized extraction materials specifically tailored for algal toxins and studies on new composite extraction materials are highly desirable. This article collects representative literature from domestic and international research on algal-toxin analysis over the past decade, summarizes the relevant findings, categorizes the applications of novel functional materials in algal-toxin-extraction processes, and provides an outlook on their future development prospects.

Closing the genome of unculturable cable bacteria using a combined metagenomic assembly of long and short sequencing reads.

Many environmentally relevant micro-organisms cannot be cultured, and even with the latest metagenomic approaches, achieving complete genomes for specific target organisms of interest remains a challenge. Cable bacteria provide a prominent example of a microbial ecosystem engineer that is currently unculturable. They occur in low abundance in natural sediments, but due to their capability for long-distance electron transport, they exert a disproportionately large impact on the biogeochemistry of their environment. Current available genomes of marine cable bacteria are highly fragmented and incomplete, hampering the elucidation of their unique electrogenic physiology. Here, we present a metagenomic pipeline that combines Nanopore long-read and Illumina short-read shotgun sequencing. Starting from a clonal enrichment of a cable bacterium, we recovered a circular metagenome-assembled genome (5.09 Mbp in size), which represents a novel cable bacterium species with the proposed name Candidatus Electrothrix scaldis. The closed genome contains 1109 novel identified genes, including key metabolic enzymes not previously described in incomplete genomes of cable bacteria. We examined in detail the factors leading to genome closure. Foremost, native, non-amplified long reads are crucial to resolve the many repetitive regions within the genome of cable bacteria, and by analysing the whole metagenomic assembly, we found that low strain diversity is key for achieving genome closure. The insights and approaches presented here could help achieve genome closure for other keystone micro-organisms present in complex environmental samples at low abundance.

Facile and fast fabrication of magnetic molecularly imprinted polymer for extraction of carbamazepine from environmental samples and fish tissues followed by UHPLC-MS/MS detection

A rapid synthesis of carbamazepine (CBZ) magnetic molecularly imprinted polymer (MMIP) was proposed based on reflux precipitation polymerization method. The synthesis time for the formation of MIP coating on MMIP was shortened to less than 4 h. The characterization of MMIP demonstrates the successful synthesis of the imprinted layer. The adsorption selectivity of MMIP was systematically evaluated and showed fast selective recognition for CBZ. The MMIP based magnetic solid-phase extraction (MSPE) combined with UHPLC-MS/MS was developed, and its feasibility was validated in four complex matrices of environmental water, sediment, fish muscle and liver. It was successfully applied for the analysis of environmental water, sediment, and different fish tissues. Four environmental water samples from different sampling sites were found to contain CBZ at concentrations < LOD or LOQ, while the sediment samples collected at the same location did not contain CBZ. Two out of three wild snakeheads were found to contain CBZ, and the liver showed the highest residue. The study of drug exposed snakeheads indicate CBZ tends to accumulate in liver, followed by brain, heart and kidneys, and finally muscle and gills. The proposed method displays high potential for analyzing CBZ in complex environmental and biological samples.

Unambiguous Analysis and Systematic Mapping of Oxygenated Aromatic Compounds in Atmospheric Aerosols Using Ultrahigh-Resolution Mass Spectrometry.

Compositional analysis of organic aerosols (OAs) at the molecular level has been a long-standing challenge in field and laboratory studies. In this work, we applied different extraction protocols to aerosol samples collected from the ambient atmosphere and biomass burning sources, followed by Orbitrap mass spectrometric analysis with a soft electrospray ionization source operating in both positive and negative ionization modes. To systematically map the distribution of mono- and dioxygenated aromatic compounds (referred to as aromatic CHO1 and CHO2 formulas) in OA, we developed a unique two-dimensional Kendrick mass defect (2D KMD) framework. Our analysis unveiled a total of (76, 64, 70) aromatic CHO1 formulas and (103, 110, 106) CHO2 formulas, corresponding to samples obtained from ambient air, rice straw burning, and sugarcane leaf burning, respectively. These results reveal a significant number of additional distinct formulas exclusively present in ambient samples, suggesting a significant chemical transformation of OAs in the atmosphere. The analytical approach can be further extended to incorporate multiple layers of 2D KMD, enabling systematic mapping of the unexplored chemical space for complex environmental samples.

PEI grafted defective MOF-808 for enhanced boron removal

Herein, we presented a facile method for enhanced boron removal based on PEI-grafted defective MOF-808 (D-MOF-808/PEI). The rich ZrOH bonds, amino-functionalized surface, and exposed metal defects of D-MOF-808/PEI endow it with the high adsorption capacity of boron. Due to the ligand exchange at metal defects and boron-imine/amine groups complexation, D-MOF-808/PEI exhibits exceptional selectivity for boron with the saturation adsorption of 95.30 mmol·g−1 at 45 °C. Additionally, negligible interference from other coexistent ions and excellent reusability promote the application of D-MOF-808/PEI practically for the rapid and high selectivity removal of boron from complex environmental samples. This work furnishes a promising strategy for boron contamination and expands the application potential of defective MOF-based adsorbents for environmental remediation.

Visual detection of microplastics using Raman spectroscopic imaging.

As a new type of pollutant in the marine environment and terrestrial ecosystems, microplastics have attracted widespread attention. Assessing the ecological risk of microplastics relies on accurately detecting small-sized particles in the environment. Microplastics exhibit unique "fingerprint" characteristics in Raman spectroscopy, making them suitable for rapid identification. In this study, we achieved visualization of microplastics through pseudo-color images generated by Raman spectroscopy imaging. Pseudo-color imaging maps were generated by selecting characteristic peaks and the classical least-squares fitting method was used to visually represent the distribution of different microplastics. The study explored the potential of Raman spectroscopy and its mapping mode in distinguishing various types of mixed microplastics and demonstrated that this approach can identify microplastics in complex environmental samples. Specifically, a cloud-point extraction followed by membrane filtration method was successfully applied to identifying mixed-component microplastics. In summary, the category, quantity, location, and differentiation of microplastics can be accurately analyzed by Raman spectroscopy, which provides a basis for assessing their ecological risk.

Ultrasensitive detection of aromatic water pollutants through protein immobilization driven organic electrochemical transistors.

Xenobiotic aromatic water pollutants pose an extreme threat to environmental sustainability. Due to the lack of detectable functional groups in these compounds and scarcity of selective bio-recognition scaffolds, easy-to-use sensing strategies capable of on-site detection remain unavailable. Herein, to address this lacune, we entail a strategy that combines biosensor scaffolds with organic electronics to create a compact device for environmental aromatic pollution monitoring. As proof of principle, a sensor module capable of rapid, economic, reliable, and ultrasensitive detection of phenol down to 2 ppb (0.02 μM) was designed wherein biosensing protein MopR was coupled with an organic electrochemical transistor (OECT). For effective interfacing of the sensing scaffold MopR, graphene oxide (GO) nanosheets were optimized as a host immobilization matrix. The MopR-GO immobilized sensor module was subsequently substituted as the gate electrode with PEDOT:PSS serving as an organic semiconductor material. The resulting OECT sensor provided a favourable microenvironment for protein activity, maintaining high specificity. Exclusive phenol detection with minimal loss of sensitivity (<5% error) could be achieved in both complex pollutant mixtures and real environmental samples. This fabrication strategy that amalgamates biological biosensors with organic electronics harnesses the potential to achieve detection of a host of emerging pollutants.

A targeted and non-targeted discovery screening approach for poly-and per-fluoroalkyl substances in model environmental biota samples

A comprehensive analytical approach for targeted and non-targeted discovery screening of per- and polyfluoroalkyl substances (PFAS) was developed and applied to model complex environmental biotic samples. Samples were extracted by formic acid-acetonitrile solution and cleaned up and fractionated by SPE (WAX). Target PFAS quantification was performed by ultra-high performance liquid chromatography interfaced with a triple quadrupole mass spectrometer (UPLC-QqQ-MS/MS). Non-targeted analysis (NTA) PFAS screening was performed with UPLC coupled with a quadrupole-Exactive orbitrap high resolution mass spectrometer (UPLC-Q-Exactive-HRMS). An iterative exclusion (IE) approach was applied to data acquisition for NTA suspect screening to increase the potential for unknown PFAS discovery with MS/MS. A complex workflow in Compound Discoverer was set up to automate data processing of the PFAS suspects search. New mass lists and MS/MS databases, which included a large number of PFAS, were set up and introduced into the search for high-throughput structure identification using HRMS techniques. The integrated targeted-NTA method successfully analyzed for legacy and alternative PFAS in model environmental biota samples, namely polar bear liver and bird egg samples. Targeted analysis provided unequivocal identification of well known/established PFAS (mainly perfluoroalkyl acids) with quantification at very low levels. The NTA suspect screening was able to determine a broader range of PFAS. The data analysis method offered high-confidence annotations for PFAS despite lacking available authentic standards. Overall, the analytical coverage of PFAS was greater and elucidated other PFAS present in these model apex predators.

Cyanobacterial harmful bloom lipopolysaccharides: pro-inflammatory effects on epithelial and immune cells in vitro

Cyanobacterial harmful blooms (CyanoHABs) pose a global ecological problem, and their lipopolysaccharides (LPS) are among the bioactive compounds they release. Previous studies on CyanoHAB-LPS from single cyanobacterial species have shown varying bioactivities in different in vitro cell models. In this study, we isolated LPS from 19 CyanoHAB samples collected at 18 water bodies in the Czech Republic over two consecutive seasons. The proportions of cyanobacteria, Gram-negative bacteria (G-), and other bacteria in the biomass were determined by qPCR, while the cyanobacterial genera were identified using light microscopy. In vitro models of keratinocytes (HaCaT), the intestinal epithelium (co-culture of differentiated Caco-2 cells and peripheral blood mononuclear cells — PBMC), and PBMC alone were treated with isolated LPS at concentrations of 50, 100, and 1 µg/ml, respectively. The endotoxin activities of these concentrations were within the range measured in the aquatic environment. Approximately 85–90% of the samples displayed biological activity. However, the potency of individual LPS effects and response patterns varied across the different in vitro models. Furthermore, the observed activities did not exhibit a clear correlation with the taxonomic composition of the phytoplankton community, the relative share of microbial groups in the biomass, endotoxin activity of the LPS, or LPS migration and staining pattern in SDS-PAGE. These findings suggest that the effects of CyanoHAB-LPS depend on the specific composition and abundance of various LPS structures within the complex environmental sample and their interactions with cellular receptors.

ScMAR-Seq: a novel workflow for targeted single-cell genomics of microorganisms using radioactive labeling.

A central question in microbial ecology is which member of a community performs a particular metabolism. Several sophisticated isotope labeling techniques are available for analyzing the metabolic function of populations and individual cells in a community. However, these methods are generally either insufficiently sensitive or throughput-limited and thus have limited applicability for the study of complex environmental samples. Here, we present a novel approach that combines highly sensitive radioisotope tracking, microfluidics, high-throughput sorting, and single-cell genomics to simultaneously detect and identify individual microbial cells based solely on their in situ metabolic activity, without prior information on community structure.

Quantitative analysis of nanoplastics in environmental and potable waters by pyrolysis-gas chromatography–mass spectrometry

Nanoplastics are emerging environmental contaminants, but their presence in environmental and potable water remains largely understudied due to the absence of quantitative analytical methods. In this study, we developed and validated a pretreatment method that combines hydrogen peroxide digestion and Amicon® Stirred Cell ultrafiltration (at 100 kDa, approximately 10 nm) with subsequent detection by pyrolysis gas chromatography–mass spectrometry (Pyr-GC/MS). This method allows for the simultaneous identification and quantification of nine selected nanoplastic types, including poly(ethylene terephthalate) (PET), polyethylene (PE), polycarbonate (PC), polypropylene (PP), poly(methyl methacrylate) (PMMA), polystyrene (PS), polyvinylchloride (PVC), nylon 6, and nylon 66, in environmental and potable water samples based on polymer-specific mass concentration. Limits of quantification ranged from 0.01 to 0.44 µg/L, demonstrating the method’s ability to quantitatively detect nanoplastics in environmental and potable water samples. Most of the selected nanoplastics were detected at concentrations of between 0.04 and 1.17 µg/L, except for PC, which was consistently below the limit of detection (<0.44 µg/L). The prevalent polymer components in the samples were PE (0.10 – 1.17 µg/L), PET (0.06 – 0.91 µg/L), PP (0.04 – 0.79 µg/L), and PS (0.06 – 0.53 µg/L) nanoplastics. The presented analytical method offers an accurate means to identify, quantify, and monitor nanoplastics in complex environmental and potable water samples. It fills gaps in our understanding of nanoplastic pollution levels, providing a valuable methodology and crucial reference data for future studies.

Development and application of a droplet digital PCR assay for the detection of Fusarium oxysporum f. sp. cubense tropical race 4 from different biological samples

AbstractFusarium wilt of banana, caused by the pathogen Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4), is a threat to food security as it affects the global banana industry. Control of this pathogen is complex due to the lack of resistant banana or plantain genotypes, lack of effective commercial fungicides, the production of chlamydospores, late appearance of symptoms in infected plants, high adaptability of the pathogen to diverse soil conditions and the polycyclic nature of the disease. Currently, the management strategy for Foc TR4 is exclusion and eradication, which is costly for the producer and the environment because the plants are treated with glyphosate and the affected area is quarantined for a long time. Thus, the development of early detection strategies for Foc TR4 from environmental samples and asymptomatic tissues with low inoculum concentration is essential; this would enable restriction of pathogen spread and minimize the environmental impact by eradication only in areas with accurate data of the pathogen's presence. In this study, three different PCR technologies (conventional PCR, quantitative PCR and droplet digital [dd] PCR) were evaluated for Foc TR4 detection in complex environmental samples. We report the development of a sensitive and specific ddPCR primer/probe set and protocols that can be used as a tool for Foc TR4 detection from symptomatic and asymptomatic plant tissue without prior DNA extraction. The limits of detection in drainage waters, footbaths and soil samples were evaluated using artificial inoculation assays. Finally, environmental samples from Foc TR4‐affected fields were analysed.

Suspect screening and effect evaluation for small-molecule agonists of the antioxidant response element pathway in fine particulate matter

Oxidative stress is an important mechanism by which fine particulate matter (PM2.5) generates toxicity. However, few inducers have been identified, and the combined effects of the contributing chemicals have rarely been examined. In this study, the occurrence of small-molecule agonists of the antioxidant response element (ARE) pathway was explored in 59 PM2.5 samples from urban Beijing over a one-year period via target and suspect screening analysis. In total, 31 chemicals with diverse structures and use categories were identified and quantified, among which polycyclic aromatic hydrocarbons (PAHs), pesticides, and phytochemicals were the most abundant chemical groups in terms of cumulative concentrations. PAHs and organonitrogen pesticides were also prioritized as the predominant contributors to the cumulative effect of ARE pathway activation, accounting for 55 % and 37 %, respectively. A combination of the prioritized chemicals (i.e., benzo[b]fluoranthene, benzo[k]fluoranthene, pyridaben, and acetochlor) was mixed at a fixed dosing ratio according to the measured average concentrations in samples, and synergism was revealed as the mode of mixture interaction. These findings highlight the importance of high-throughput chemical screening for identifying hazardous components in complex environmental samples, and also expand the knowledge regarding the components contributing to PM2.5-induced oxidative stress.