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.

The high density and viscosity of fuel oil leads to its prolonged persistence in the environment and causes widespread contamination. Dispersants with a low environmental impact are necessary for fuel oil spill remediation. This study aimed to formulate bio-based dispersants by mixing anionic biosurfactant (lipopeptides from Bacillus subtilis GY19) with nonionic oleochemical surfactant (Dehydol LS7TH). The synergistic effect of the anionic-nonionic surfactant mixture produced a Winsor Type III microemulsion, which promoted petroleum mobilization. The hydrophilic-lipophilic deviation (HLD) equations for ionic and nonionic surfactant mixtures were compared, and it was found that the ionic equation was applicable for the calculation of lipopeptides and Dehydol LS7TH concentrations. The best formula contained 6.6% w/v lipopeptides and 11.9% w/v Dehydol LS7TH in seawater, and its dispersion effectiveness for bunker fuels A and C was 92% and 78%, respectively. The application of bio-based dispersants in water sources was optimized by Box-Behnken design. The efficiency of the bio-based dispersant was affected by the dispersant-to-oil ratios (DORs) but not by the water salinity. A suitable range of DORs for different oil contamination levels could be identified from the response surface plot. The dispersed fuel oil was further degraded by adding an oil-degrading bacterial consortium to the chemically enhanced water accommodated fractions (CEWAFs). After 7 days of incubation, the concentration of fuel oil was reduced from 3692 mg/L to 356 mg/L (88% removal efficiency). On the other hand, the abiotic control removed less than 40% fuel oil from the CEWAFs. This bio-based dispersant had an efficiency comparable to that of a commercial dispersant. The process of dispersant formulation and optimization could be applied to other surfactant mixtures.

Bacteria–phyllosilicate complexes are commonly found in natural environments and are capable of immobilizing trace metals. However, the molecular binding mechanisms of heavy metals to these complex aggregates still remain poorly understood. This study investigated Cd adsorption on Gram-positive Bacillus subtilis, Gram-negative Pseudomonas putida and their binary mixtures with montmorillonite using surface complexation model, Cd K-edge extended X-ray absorption fine structure (EXAFS) spectroscopy and isothermal titration calorimetry (ITC). We have shown that larger amounts of Cd are adsorbed by B. subtilis than by P. putida at pH<∼6, and Cd sorption that binding to phosphate groups plays a more important role in P. putida than in B. subtilis. This remind us that we should consider the microbe species when predict the biochemical behavior of trace metals in microbe-bearing environments. The observed Cd adsorption on the binary bacteria–clay composites was more than that predicted based on the component additivity approach. When taking bacteria–clay (1:1 mass ratio) as a representative example, an approximately 68%:32% metal distribution between the bacterial and mineral fraction was found. Both the EXAFS and ITC fits showed that the binding stoichiometry for Cd-carboxyl/phosphate was smaller in the binary mixtures than that in pure bacteria. We proposed that the significant deviations were possibly due to the physical-chemical interaction between the composite fractions that might reduce the agglomeration of the clay grains, increase the negative surface charges, and provide additional bridging of metals ions between bacterial cells and clays.

Colloids (non-biological and biological) with different sizes are ubiquitous in natural environment. The investigations regarding the influence of different-sized colloids on the transport and deposition behaviors of engineered-nanoparticles in porous media yet are still largely lacking. This study investigated the effects of different-sized non-biological and biological colloids on the transport of titanium dioxide nanoparticles (nTiO2) in quartz sand under both electrostatically favorable and unfavorable conditions. Fluorescent carboxylate-modified polystyrene latex microspheres (CML) with sizes of 0.2–2 μm were utilized as model non-biological colloids, while Gram-negative Escherichia coli (∼1 μm) and Gram-positive Bacillus subtilis (∼2 μm) were employed as model biological colloids. Under the examined solution conditions, both breakthrough curves and retained profiles of nTiO2 with different-sized CML particles/bacteria were similar as those without colloids under favorable conditions, indicating that the copresence of model colloids in suspensions had negligible effects on the transport and deposition of nTiO2 under favorable conditions. In contrast, higher breakthrough curves and lower retained profiles of nTiO2 with CML particles/bacteria relative to those without copresent colloids were observed under unfavorable conditions. Clearly, the copresence of model colloids increased the transport and decreased the deposition of nTiO2 in quartz sand under unfavorable conditions (solution conditions examined in present study). Both competition of deposition sites on quartz sand surfaces and the enhanced stability/dispersion of nTiO2 induced by copresent colloids were found to be responsible for the increased nTiO2 transport with colloids under unfavorable conditions. Moreover, the smallest colloids had the highest coverage on sand surface and most significant dispersion effect on nTiO2, resulting in the greatest nTiO2 transport.

A pyrene-degrading consortium OPK containing Mycolicibacterium strains PO1 and PO2, Novosphingobium pentaromativorans PY1 and Bacillus subtilis FW1 effectively biodegraded medium- and long-chain alkanes as well as mixed hydrocarbons in crude oil. The detection of alkB and CYP153 genes in the genome of OPK members supports its phenotypic ability to effectively degrade a broad range of saturated hydrocarbons in crude oil. Zeolite-immobilized OPK was developed as a ready-to-use bioproduct and it exhibited 74% removal of 1000 mg L⁻¹ crude oil within 96 h in sterilized seawater without nutrient supplementation and maintained high crude oil-removal activity under a broad range of pH values (5.0–9.0), temperatures (30–40 °C) and salinities (20–60‰). In addition, the immobilized OPK retained a high crude oil removal efficacy in semicontinuous experiments and showed reusability for at least 5 cycles. Remarkably, bioaugmentation with zeolite-immobilized OPK in sandy soil microcosms significantly increased crude oil (10,000 mg kg⁻¹ soil) removal from 45% to 80.67% within 21 days compared to biostimulation and natural attenuation. Moreover, bioaugmentation with exogenous immobilized OPK stimulated an increase in the relative abundances of Alcanivorax genus, indigenous hydrocarbon-degrading bacteria, which in turn enhanced removal efficiency of crude oil contamination from sandy soil microcosms. The results indicate positive interactions between the bioaugmented immobilized consortium, harboring Mycolicibacterium as a key player, and indigenous Alcanivorax, which exhibited crucial functions for improving crude oil removal efficacy. The knowledge obtained forms an important basis for further synthesis and handling of a promising bio-based product for enhancing the in situ bioremediation of crude oil-polluted marine environments.

The present study showed that syntrophic associations in a defined bacterial consortium, named OPK, containing Mycolicibacterium strains PO1 and PO2, Novosphingobium pentaromativorans PY1 and Bacillus subtilis FW1, led to effective pyrene degradation over a wide range of pH values, temperatures and salinities, as well as in the presence of a second polycyclic aromatic hydrocarbon (PAH). Anthracene, phenanthrene or fluorene facilitated complete pyrene degradation within 9 days, while fluoranthene delayed pyrene degradation. Interestingly, fluoranthene degradation was enhanced in the presence of pyrene. Transcriptome analysis confirmed that Mycolicibacterium strains were the key PAH-degraders during the cometabolism of pyrene and fluoranthene. Notably, the transcription of genes encoding pyrene-degrading enzymes were shown to be important for enhanced fluoranthene degradation. NidAB was the major initial oxygenase involved in the degradation of pyrene and fluoranthene mixture. Other functional genes encoding ribosomal proteins, an iron transporter, ABC transporters and stress response proteins were induced in strains PO1 and PO2. Furthermore, an intermediate pyrene-degrading Novosphingobium strain contributed to protocatechuate degradation. The results demonstrated that synergistic interactions among the bacterial members (PO1, PO2 and PY1) of the consortium OPK promoted the simultaneous degradation of two high molecular weight (HMW) PAHs.

This work aimed to study the performance of paraquat removal by cell-immobilized ceramics. Two strains of paraquat degrading bacteria, Pseudomonas putida and Bacillus subtilis, were separately immobilized on the ceramic with and without wastewater sludge addition. Results showed that the ceramic surface with sludge has more functional groups and a more highly negative charge on the surface than the original ceramic. The ceramic with sludge had 2-3-fold of the immobilized cells higher than that of the control (without sludge) and less leaching of the immobilized cells. The sludge addition at 20% (w/w) to the ceramic provided the highest cell adhesion for both P. putida and B. subtilis. The paraquat removal efficiencies were higher than 98%, while the control ceramic could remove only 77 ± 1.2%. The immobilized cells on ceramic with sludge provided a significant degree of dissolved organic nitrogen reduction (82%) during the paraquat removal. Most organic nitrogen in paraquat was biologically mineralized (ammonified). Findings from this work suggest the superiority of ceramic with sludge in mineralizing organic nitrogen associated with paraquat.

Phthalate ester pollution in the environment and food chain is frequently reported. Microbial treatment is a green and efficient method for solving this problem. The isolation and systematic investigation of microorganisms generally recognized as safe (GRAS) will provide useful resources. A GRAS Bacillus subtilis strain, BJQ0005, was isolated from Baijiu fermentation starter and efficiently degraded phthalate esters (PAEs). The half-lives for di-isobutyl phthalate, di-butyl phthalate and di-(2-ethylhexyl) phthalate were 3.93, 4.28, and 25.49 h, respectively, from the initial amount of 10 mg per 10 mL reaction mixture, which are records using wild-type strains. Genome sequencing and metabolic intermediate analysis generated the whole metabolic pathway. Eighteen enzymes from the α/β hydrolase family were expressed. Enzymes GTW28_09400 and GTW28_13725 were capable of single ester bond hydrolysis of PAEs, while GTW28_17760 hydrolyzed di-ester bonds of PAEs. Using molecular docking, a possible mechanism affecting enzymatic ester bond hydrolysis of mono-butyl phthalate was proposed of GTW28_17760. The carboxyl group generated by the first hydrolysis step interacted with histidine in the catalytic active center, which negatively affected enzymatic hydrolysis. Isolation and systematic investigation of the PAE degradation characteristics of B. subtilis will promote the green and safe treatment of PAEs in the environment and food industry.

Bacillus subtilis YB1 is a strain that can efficiently transform nicosulfuron. In order to study its remediation ability and effects on other microorganisms in the soil, indoor biological remediation experiments and rhizosphere microbial diversity analysis were performed. B. subtilis YB1 granules were prepared and applied to the nicosulfuron contaminated soil. The concentration of nicosulfuron was detected by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and changes in the physiological indicators of wheat were measured. At the same time, the changes in the rhizosphere soil microbial diversity were determined by 16S RNA sequencing. Results showed that the YB1 granules made a contribution to the transformation of nicosulfuron (0.05 mg kg⁻¹) in the soil within 55 days. The physiological indicators of wheat also showed consistent result about nicosulfuron transformation. Rhizosphere soil microbial diversity results indicated the relative abundance of Firmicutes decreased (3.0%–0.35%) and Acidobacteria first decreased (25.82%–22.38%) and then increased (22.3%–26.1%) with nicosulfuron added (N group). The relative abundance of Acidobacteria first decreased (25.8%–15.3%) and then increased (15.3%–21.7%) while Proteobacteria increased (26.5%–38.08%). At the same time, Firmicutes first increased (2.6%–12.3%) and then decreased to original level (12.3%–0.7%) in the N group with YB1 granules (NYB1 group). Members of the genus Bacillus initially increased and then decreased to the original level as the Control group, therefore, they did not become dominant in the rhizosphere soil. Alpha diversity analyses showed no obvious differences in species diversity among the N, NYB1 and Control groups. So YB1 did not have obvious influence on the rhizosphere microbial community structure during nicosulfuron transformation, which only had some effect on species abundance. This study revealed the successful indoor bioremediation of nicosulfuron in the soil, providing a potential strategy for solving the problem of nicosulfuron contamination.

In this work, soil contaminated by petroleum resins was remediated by electrokinetic-bioremediation (EK-BIO) technology for 60 days. A microbial consortium, comprising Rhizobium sp., Arthrobacter globiformis, Clavibacter xyli, Curtobacterium flaccumfaciens, Bacillus subtilis, Pseudomonas aeruginosa and Bacillus sp., was used to enhance the treatment performance. The results indicate that resin removal and phytotoxicity reduction were highest in the inoculated EK process, wherein 23.6% resins was removed from the soil and wheat seed germination ratio was increased from 47% to around 90% after treatment. The microbial counts, soil basal respiration and dehydrogenase activity were positively related to resins degradation, and they could be enhanced by direct current electric field. After remediation, the C/H ratio of resins decreased from 8.03 to 6.47. Furthermore, the structure of resins was analyzed by Fourier-transform infrared spectroscopy, elemental analysis, and ¹H nuclear magnetic resonance (¹H NMR) before and after treatment. It was found that the changes of the structure of resins took place during EK-BIO treatment and finally led to the reduction of aromaticity, aromaticity condensation and phytotoxicity.