In this study, we treated harmful Microcystis aeruginosa cyanobacteria using chitosan-modified nanobubbles. The chitosan-modified nanobubbles (255 ± 19 nm) presented a positive zeta potential (15.36 ± 1.17 mV) and generated significantly (p < 0.05) more hydroxyl radicals than the negatively charged nanobubbles (−20.68 ± 1.11 mV). Therefore, the interaction between the positively charged chitosan-modified nanobubbles and negatively charged M. aeruginosa (−34.81 ± 1.79 mV) was favored. The chitosan-modified nanobubble treatment (2.20 × 10⁸ particles mL⁻¹) inactivated 73.16% ± 2.23% of M. aeruginosa (2.00 × 10⁶ cells mL⁻¹) for 24 h without causing significant cell lysis (≤0.25%) and completely inhibited the acute toxicity of M. aeruginosa toward Daphnia magna. The inactivation was correlated (r² = 0.97) with the formation of reactive oxygen species (ROS) in M. aeruginosa. These findings indicated that the hydroxyl radicals generated by the chitosan-modified nanobubbles disrupted cell membrane integrity and enhanced oxidative stress (ROS formation), thereby inactivating M. aeruginosa. Moreover, the penetration of the chitosan-modified nanobubbles and cell alterations in M. aeruginosa were visually confirmed. Our results suggested that the chitosan-modified nanobubble treatment is an eco-friendly method for controlling harmful algae. However, further studies are required for expanding its practical applications.

Discharged carbon nanotubes (CNTs) likely interact with co-existing organic contaminants (OCs) and pose joint toxicity to environmental microbes. Herein, hydrophobic pentachlorophenol (PCP) and hydrophilic ciprofloxacin (CIP) were used as representative OCs and their joint toxicities with CNTs to Bacillus subtilis were systematically investigated at cellular, biochemical, and omics levels. The 3-h bacterial growth half inhibitory concentrations of CNTs, PCP, and CIP were 12.5 ± 2.6, 3.5 ± 0.5, and 0.46 ± 0.03 mg/L, respectively, and they all could damage cell membrane, increase intracellular oxidative stress, and alter bacterial metabolomics and transcriptomics; while CNTs-PCP and CNTs-CIP binary exposures exhibited distinct additive and synergistic toxicities, respectively. CNTs increased bacterial bioaccumulation of PCP and CIP via destabilizing and damaging cell membrane. PCP reduced the bioaccumulation of CNTs, while CIP had no significant effect; this difference could be owing to the different effects of the two OCs on cell-surface hydrophobicity and CNTs electronegativity. The additive toxicity outcome upon CNTs-PCP co-exposure could be a result of the balance between the increased toxicity from increased PCP bioaccumulation and the decreased toxicity from decreased CNTs bioaccumulation. The increased bioaccumulation of CIP contributed to the synergistic toxicity upon CNTs-CIP co-exposure, as confirmed by the increased inhibition of topoisomerase Ⅳ activity and interference in gene expressions regulating ABC transporters and lysine biosynthesis. The findings provide novel insights into environmental risks of CNTs.

Plastic materials are increasingly produced worldwide with a total estimated production of >8300 million tonnes to date, of which 60% was discarded. In the environment, plastics fragment into smaller particles, e.g. microplastics (size < 5 mm), and further weathering leads to the formation of functionally different contaminants – nanoplastics (size <1 μm). Nanoplastics are believed to have entirely different physical (e.g. transport), chemical (e.g. functional groups at the surface) and biological (passing the cell membrane, toxicity) properties compared to the micro- and macroplastics, yet, their measurement in the environmental samples is seldom available. Here, we present measurements of nanoplastics mass concentration and calculated the deposition at the pristine high-altitude Alpine Sonnblick observatory (3106 MASL), during the 1.5 month campaigh in late winter 2017. The average nanoplastics concentration was 46.5 ng/mL of melted surface snow. The main polymer types of nanoplastics observed for this site were polypropylene (PP) and polyethylene terephthalate (PET). We measured significantly higher concentrations in the dry sampling periods for PET (p < 0.002) but not for PP, which indicates that dry deposition may be the preferential pathway for PET leading to a gradual accumulation on the snow surfaces during dry periods. Air transport modelling indicates regional and long-range transport of nanoplastics, originating preferentially from European urban areas. The mean deposition rate was 42 (+32/-25) kg km⁻² year⁻¹. Thus more than 2 × 10¹¹ nanoplastics particles are deposited per square meter of surface snow each week of the observed period, even at this remote location, which raises significant toxicological concerns.

Hydrophobic microplastics with a relatively large surface area can act as carriers for pollutants and exert a series of indirect effects on crop plants. This study investigated the toxic effects of small polystyrene (SPS, 100–1000 nm) and large polystyrene (LPS, >10,000 nm) microplastics, on lettuce under di-butyl phthalate (DBP) stress. The results indicated that single SPS, LPS, and DBP treatments significantly decreased lettuce biomass, and induced oxidative stress and damaged lettuce leaves and roots. According to Gaussian analysis, SPS or LPS could interact with DBP through van der Waals force, thereby reducing lettuce biomass and DBP enrichment in roots and leaves under combined treatments, increasing antioxidant enzyme activities and exacerbating oxidative stress and subcellular damage, compared to single DBP treatments. Observation using scanning electron microscopy demonstrated that polystyrene (PS) adhered to the root surfaces, which, in turn, caused physical blockage of the root pores. Cell membrane and wall damage was observed during PS and/or DBP exposures, as identified by transmission electron microscopy. Molecular docking illustrated that DBP and monobutyl phthalate could interact with superoxide dismutase residues through hydrogen bonding, π-π stacking, alkyl conjugation, and van der Waals forces. Interestingly, there were no statistical differences between the phytotoxicity of nano- and microplastics to lettuce. These findings showed that PS aggravated DBP-induced phytotoxicity.

The surface modifications of nanoparticles (NPs), are well-recognized parameters that affect the toxicity, while there has no study on toxicity of Al₂O₃ NPs with different surface modification. Therefore, for the first time, this study pays attention to evaluating the toxicity and potential mechanism of pristine Al₂O₃ NPs (p-Al₂O₃), hydrophilic (w-Al₂O₃) and lipophilic (o-Al₂O₃) modifications of Al₂O₃ NPs both in vitro and in vivo. Applied concentrations of 10, 20, 40, 80,100 and 200 μg/mL for 24 h exposure on Caenorhabditis elegans (C. elegans), while 100 μg/mL of Al₂O₃ NPs significantly decreased the survival rate. Using multiple toxicological endpoints, we found that o-Al₂O₃ NPs (100 μg/mL) could induce more severe toxicity than p-Al₂O₃ and w-Al₂O₃ NPs. After uptake by C. elegans, o-Al₂O₃ NPs increased the intestinal permeability, easily swallow and further destroy the intestinal membrane cells. Besides, cytotoxicity evaluation revealed that o-Al₂O₃ NPs (100 μg/mL) are more toxic than p-Al₂O₃ and w-Al₂O₃. Once inside the cell, o-Al₂O₃ NPs could attack mitochondria and induce the over-production of reactive oxygen species (ROS), which destroy the intracellular redox balance and lead to apoptosis. Furthermore, the transcriptome sequencing and RT-qPCR data also demonstrated that the toxicity of o-Al₂O₃ NPs is highly related to the damage of cell membrane and the imbalance of intracellular redox. Generally, our study has offered a comprehensive sight to the adverse effects of different surface modifications of Al₂O₃ NPs on environmental organisms and the possible underlying mechanisms.

Wildfires are a complex environmental problem worldwide. The ashes produced during the fire bear metals and PAHs with high toxicity and environmental persistence. These are mobilized into downhill waterbodies, where they can impair water quality and human health. In this context, the present study aimed at assessing the toxicity of mimicked wildfire runoff to human skin cells, providing a first view on the human health hazardous potential of such matrices. Human keratinocytes (HaCaT) were exposed to aqueous extracts of ashes (AEA) prepared from ash deposited in the soil after wildfires burned a pine or a eucalypt forest stand. Cytotoxicity (MTT assay) and changes in cell cycle dynamics (flow cytometry) were assessed. Cell viability decreased with increasing concentrations of AEA, regardless of the ash source, the extracts preparation method (filtered or unfiltered to address the dissolved or the total fractions of contaminants, respectively) or the exposure period (24 and 48 h). The cells growth was also negatively affected by the tested AEA matrices, as evidenced by a deceleration of the progress through the cell cycle, namely from phase G0/G1 to G2. The cytotoxicity of AEA could be related to particulate and dissolved metal content, but the particles themselves may directly affect the cell membrane. Eucalypt ash was apparently more cytotoxic than pine ash due to differential ash metal burden and mobility to the water phase. The deceleration of the cell cycle can be explained by the attempt of cells to repair metal-induced DNA damage, while if this checkpoint and repair pathways are not well coordinated by metal interference, genomic instability may occur. Globally, our results trigger public health concerns since the burnt areas frequently stand in slopes of watershed that serve as recreation sites and sources of drinking water, thus promoting human exposure to wildfire-driven contamination.

The synergistic cooperation of microbial cells and their extracellular polymeric substances (EPS) in biofilms is critical for the biofilm’s resistance to heavy metals and the migration and transformation of heavy metals. However, the effects of different components of biofilms have not been fully understood. In this study, the spatial distribution and speciation of copper in the colloidal EPS, capsular EPS, cell walls and membranes, and intracellular fraction of unsaturated Pseudomonas putida (P. putida) CZ1 biofilms were fully determined at the subcellular level. It was found that 60–67% of copper was located in the extracellular fraction of biofilms, with 44.7–42.3% in the capsular EPS. In addition, there was 15.5–20.1% and 17.2–21.2% of copper found in the cell walls and membranes or the intracellular fraction, respectively. Moreover, an X-ray absorption fine structure spectra analysis revealed that copper was primarily bound by carboxyl-, phosphate-, and hydrosulfide-like ligands within the extracellular polymeric matrix, cell walls and membranes, and intracellular fraction, respectively. In addition, macromolecule quantification, fourier-transform infrared spectroscopy spectra and sulfur K-edge x-ray absorption near edge structure analysis further showed the carboxyl-rich acidic polysaccharides in EPS, phospholipids in cell walls and cell membranes, and thiol-rich intracellular proteins were involved in binding of copper in the different components of biofilm. The full understanding of the distribution and chemical species of heavy metals in biofilms not only promotes a deep understanding of the interaction mechanisms between biofilms and heavy metals, but also contributes to the development of effective biofilm-based heavy metal pollution remediation technologies.

Fungi have an exceptional capability to flourish in presence of heavy metals and pesticide. However, the mechanism of bioremediation of pesticide (lindane) and multimetal [mixture of cadmium (Cd), chromium (Cr), copper (Cu), nickel (Ni), lead (Pb), zinc (Zn)] by a fungus is little understood. In the present study, Aspergillus fumigatus, a filamentous fungus was found to accumulate heavy metals in the order [Zn(98%)>Pb(95%)>Cd(63%)>Cr(62%)>Ni(46%)>Cu(37%)] from a cocktail of 30 mg L⁻¹ multimetal and lindane (30 mg L⁻¹) in a composite media amended with 1% glucose. Particularly, Pb and Zn uptake was enhanced in presence of lindane. Remarkably, lindane was degraded to 1.92 ± 0.01 mg L⁻¹ in 72 h which is below the permissible limit value (2.0 mg L⁻¹) for the discharge of lindane into the aquatic bodies as prescribed by European Community legislation. The utilization of lindane as a cometabolite from the complex environment was evident by the phenomenal growth of the fungal pellet biomass (5.89 ± 0.03 g L⁻¹) at 72 h with cube root growth constant of fungus (0.0211 g¹/³ L⁻¹/³ h⁻¹) compared to the biomasses obtained in case of the biotic control as well as in presence of multimetal complex without lindane. The different analytical techniques revealed the various stress coping strategies adopted by A. fumigatus for multimetal uptake in the simultaneous presence of multimetal and pesticide. From the Transmission electron microscope coupled energy dispersive X-ray analysis (TEM-EDAX) results, uptake of the metals Cd, Cu and Pb in the cytoplasmic membrane and the accumulation of the metals Cr, Ni and Zn in the cytoplasm of the fungus were deduced. Fourier-transform infrared spectroscopy (FTIR) revealed involvement of carboxyl/amide group of fungal cell wall in metal chelation. Thus A. fumigatus exhibited biosorption and bioaccumulation as the mechanisms involved in detoxification of multimetals.

The increase of CO₂ concentration in the atmosphere, water and soil environment can lead to the changes in microbial activities. However, the transformation of antibiotic resistance genes has not been investigated in the presence of higher levels of CO₂. This study demonstrated that CO₂ facilitated the transformation of pUC19 plasmid, carrying ampicillin resistance genes, into Escherichia coli. Mechanism studies revealed that the type Ⅱ secretion system, type Ⅳ pilus and some other secretion systems were enhanced by CO₂, leading to DNA capture by pilus, larger cell pore sizes and more cell membrane channels. CO₂ also increased reactive oxygen species production, leading to SOS response and cell membrane damage. Besides, changes in intracellular Fe²⁺ and Mg²⁺ concentrations induced by CO₂ caused greater damage to the cell membrane and enhanced secretion systems, respectively. Overall, increased CO₂ provided more cell membrane channels for plasmid uptake and led to higher transformation efficiencies. The potential risk of a natural factor on the transformation of ARGs was first studied in this study, which helps us understand the fate of ARGs in ecosystems. As the carbon emission will continue to grow and enhance the enrichment of CO₂ in water and soil, the findings revealed a more severe public health issue under the background of carbon emission and CO₂ leakage.

Phycoremediation technologies significantly contribute to solving serious problems induced by heavy metals accumulation in the aquatic systems. Here we studied the mechanisms underlying Al stress tolerance in two diazotrophic cyanobacterial species, to identify suitable species for Al phycoremediation. Al uptake as well as the physiological and biochemical responses of Anabaena laxa and Nostoc muscorum to 7 days Al exposure at two different concentrations i.e., mild (100 μM) and high dose (200 μM), were investigated. Our results revealed that A. laxa accumulated more Al, and it could acclimatize to long-term exposure of Al stress. Al induced a dose-dependent decrease in photosynthesis and its related parameters e.g., chlorophyll content (Chl a), phosphoenolpyruvate carboxylase (PEPC) and Ribulose‒1,5‒bisphosphate carboxylase/oxygenase (RuBisCo) activities. The affect was less pronounced in A. laxa than N. muscorum. Moreover, Al stress significantly increased cellular membrane damage as indicated by induced H₂O₂, lipid peroxidation, protein oxidation, and NADPH oxidase activity. However, these increases were lower in A. laxa compared to N. muscorum. To mitigate the impact of Al stress, A. laxa induced its antioxidant defense system by increasing polyphenols, flavonoids, tocopherols and glutathione levels as well as peroxidase (POX), catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GPX) enzymes activities. On the other hand, the antioxidant increases in N. muscorum were only limited to ascorbate (ASC) cycle. Overall, high biosorption/uptake capacity and efficient antioxidant defense system of A. laxa recommend its feasibility in the treatment of Al contaminated waters/soils.