Butachlor (BUT) is a chloroacetanilide herbicide widely applied to rice paddies to control annual grass and broad-leaf weeds. A BUT-degrading bacterial strain (PK) was isolated from paddy soils. Biochemical and 16S rRNA sequencing characteristics confirmed the strain as Pseudomonas aeruginosa (99% resemblance). The isolate dissipated BUT (100 μg/mL) in an M9 liquid medium with a rate of 0.5 ± 0.03 day-1 and DT50 and DT90 of 1.38 ± 0.10 days and 4.58 ± 0.32 days, respectively. Soil dissipation of BUT was investigated under flooded conditions. In sterile soils, the isolate increased the dissipation of BUT (200 μg/g) (DT50 = 12.38 ± 1.83 days, DT90 = 41.12 ± 6.09 days, k = 0.06 ± 0.01 day-1) compared to sterile non-inoculated samples (DT50 = 26.87 ± 2.82 days, DT90 = 89.25 ± 9.36 days, k = 0.03 ± 0.00 day-1). In non-inoculated non-sterile soil experiments, the dissipation of BUT was faster (DT50 = 15.17 ± 2.11 days, DT90 = 50.38 ± 7.02 days, k = 0.05 ± 0.00 day-1) compared to non-inoculated sterile ones, and inoculating the isolate accelerated the removal of BUT in non-sterile soils significantly (DT50 = 8.03 ± 1.20 days, DT90 = 26.68 ± 3.97 days, k = 0.09 ± 0.01 day-1). BUT inhibited soil respiration (SR) initially for 5 days, followed by an increase until day 20. The increase in SR was more pronounced in the co-presence of BUT and the isolate. The results of this research suggest P. aeruginosa PK as a suitable candidate for BUT bioremediation.

International audience | This study focused on the role of bioaccessibility in the phenanthrene (PHE) biodegradation in diffusely contaminated soil, by combining chemical and microbiological approaches.First, we determined PHE dissipation rates and PHE sorption/desorption isotherms for two soils (PPY and Pv) presenting similar chronic PAH contamination, but different physico-chemical properties.Our results revealed that the PHE dissipation rate was significantly higher in the Pv soil compared to the PPY soil, while PHE sorption/desorption isotherms were similar. Interestingly, increases of PHE desorption and potentially of PHE bioaccessibility were observed for both soils when adding rhamnolipids (biosurfactants produced by Pseudomonas aeruginosa). Second, using C-13-PHE incubated in the same soils, we analyzed the PHE degrading bacterial communities. The combination of stable isotope probing (DNA-SIP) and 16S rRNA gene pyrosequencing revealed that Betaproteobacteria were the main PHE degraders in the Pv soil, while a higher bacterial diversity (Alpha-, Beta-, Gammaproteobacteria and Actinobacteria) was involved in PHE degradation in the PPY soil. The amendment of biosurfactants commonly used in biostimulation methods (i.e. rhamnolipids) to the two soils clearly modified the PHE sorption/desorption isotherms, but had no significant impact on PHE degradation rates and PHE-degraders identity.These results demonstrated that increasing the bioaccessibility of PHE has a low impact on its degradation and on the functional populations involved in this degradation.

Noble metal–based nanomaterials (NMNs), such as platinum nanoparticles (Pt@NPs) and palladium nanoparticles (Pd@NPs), are increasingly being used as antibacterial agents. However, little information is available on bacterial resistance to NMNs. In this study, owing to their oxidase-like and peroxidase-like properties, both Pt@NPs and Pd@NPs induce reactive oxygen species (ROS) and manifest antibacterial activities: 6.25 μg/mL of either Pt@NPs or Pd@NPs killed >50% of Staphylococcus aureus strain ATCC29213. However, Pseudomonas aeruginosa strain PAO1 completely resisted 12.5 μg/mL of Pt@NPs and 6.25 μg/mL of Pd@NPs. Compared to the non-NMN groups, these NMNs promoted 2–3-fold upregulation of the quorum sensing (QS) gene lasR in strain PAO1. In fact, the lasR gene upregulation induced a 1.5-fold reduction in ROS production and increased biofilm formation by 11% (Pt@NPs) and 27% (Pd@NPs) in strain PAO1. The ΔlasR mutants (lasR gene knock out in strain PAO1), became sensitive to NMNs. The survival rates of ΔlasR mutants at 12.5 μg/mL Pt@NPs and Pd@NPs treatments were only 77% and 58%, respectively. This is the first report indicating that bacteria can resist NMNs through QS. Based on these results, evaluation of the ecological risks of using NMNs as antibacterial agents is necessary.

Most chemical plant wastewater contains both organic and inorganic pollutants, which are easy to diffuse along with surface runoff. The combined pollution of nonylphenol (NP) and cadmium (Cd) in soil is a serious problem that has not attracted enough attention. Based on the effects of selenium (Se) and Pseudomonas aeruginosa (P. aeruginosa) on plant and soil microbial communities, we speculated that the application of Se and P. aeruginosa in soil could improve the phytoremediation efficiency of ryegrass on contaminated soil. In this study, pot experiments with Cd and NP co-contaminated soil were conducted, and the results showed that application of P. aeruinosa alone could improve the removal rates of NP and Cd by ryegrass, and the supplementary of Se further enhanced the effect of micro-phyto remediation, with the highest removal rates of NP and Cd were 79.6% and 49.4%, respectively. The application of P. aeruginosa plus Se reduced the adsorption of Cd and NP through C–O and Si–O–Fe of the soil, changed the enzyme activity, and also affected the changing trend of the microbial community in soil. Pseudomonas, Sphingomonadales, Nitrospira, and other beneficial bacteria were enriched after a 60-day period with P. aeruginosa and Se treatment, thus promoting the removal of NP and Cd. In light of the above results, we suggest that P. aeruginosa application can efficiently facilitate the phytoremediation of ryegrass on Cd-NP co-contaminated soil, and Se supplementation in soil showed the synergistic effect on the remediation.

Biodiesel side stream waste glycerol was identified as a cheap carbon source for rhamnolipids (RLs) production which at the same time could improve the management of waste. The present study aimed to produce RLs by using Pseudomonas aeruginosa RS6 utilizing waste glycerol as a substrate and to evaluate their physico-chemicals properties. Fermentation conditions such as temperature, initial medium pH, waste glycerol concentration, nitrogen sources and concentrations resulted in different compositions of the mono- and di-RLs produced. The maximum RLs production of 2.73 g/L was obtained when P. aeruginosa RS6 was grown in a basal salt medium supplemented with 1% waste glycerol and 0.2 M sodium nitrate at 35 °C and pH 6.5. At optimal fermentation conditions, the emulsification index (E₂₄) values of cooking oil, diesel oil, benzene, olive oil, petroleum, and kerosene were all above E₂₄₌50%. The surface tension reduction obtained from 72.13 mN/m to 29.4–30.4 mN/m was better than the surface activity of some chemical-based surfactants. The RLs produced possessed antimicrobial activities against Gram-negative and Gram-positive bacteria with values ranging from 37% to 77% of growth inhibition when 1 mg/mL of RLs was used. Concentrations of RLs below 1500 μg/mL did not induce phytotoxicity effects on the tested seeds (Vigna radiata) compared to the chemical-based- surfactant, SDS. Furthermore, RLs tested on zebrafish (Danio rerio) embryos only exhibited low acute toxicity with an LC₅₀ value of 72.97 μg/mL at 48 h of exposure suggesting a green and eco-biochemical worthy of future applications to replace chemical-based surfactants.

Dibenzofuran (DBF) derivatives have caused serious environmental problems, especially those produced by paper pulp bleaching and incineration processes. Prominent for its resilient mutagenicity and toxicity, DBF poses a major challenge to human health. In the present study, a new strain of Pseudomonas aeruginosa, FA-HZ1, with high DBF-degrading activity was isolated and identified. The determined optimum conditions for cell growth of strain FA-HZ1 were a temperature of 30 °C, pH 5.0, rotation rate of 200 rpm and 0.1 mM DBF as a carbon source. The biochemical and physiological features as well as usage of different carbon sources by FA-HZ1 were studied. The new strain was positive for arginine double hydrolase, gelatinase and citric acid, while it was negative for urease and lysine decarboxylase. It could utilize citric acid as its sole carbon source, but was negative for indole and H2S production. Intermediates of DBF 1,2-dihydroxy-1,2-dihydrodibenzofuran, 1,2-dihydroxydibenzofuran, 2-hydroxy-4-(3′-oxo-3′H-benzofuran-2′-yliden)but-2-enoic acid, 2,3-dihydroxybenzofuran, 2-oxo-2-(2′-hydrophenyl)lactic acid, and 2-hydroxy-2-(2′-hydroxyphenyl)acetic acid were detected and identified through liquid chromatography-mass analyses. FA-HZ1 metabolizes DBF by both the angular and lateral dioxygenation pathways. The genomic study identified 158 genes that were involved in the catabolism of aromatic compounds. To identify the key genes responsible for DBF degradation, a proteomic study was performed. A total of 1459 proteins were identified in strain FA-HZ1, of which 100 were up-regulated and 104 were down-regulated. A novel enzyme “HZ6359 dioxygenase”, was amplified and expressed in pET-28a in E. coli BL21(DE3). The recombinant plasmid was successfully constructed, and was used for further experiments to verify its function. In addition, the strain FA-HZ1 can also degrade halogenated analogues such as 2, 8-dibromo dibenzofuran and 4-(4-bromophenyl) dibenzofuran. Undoubtedly, the isolation and characterization of new strain and the designed pathways is significant, as it could lead to the development of cost-effective and alternative remediation strategies. The degradation pathway of DBF by P. aeruginosa FA-HZ1 is a promising tool of biotechnological and environmental significance.

High abundances of antibiotic-resistant pathogenic bacteria (ARPB) and antibiotic resistance genes (ARGs) in agricultural soil-plant systems have become serious threats to human health and environmental safety. Therefore, it is crucial to develop targeted technology to control existing antibiotic resistance (AR) contamination and potential dissemination in soil-plant systems. In this work, polyvalent bacteriophage (phage) therapy and biochar amendment were applied separately and in combination to stimulate ARPB/ARG dissipation in a soil-lettuce system. With combined application of biochar and polyvalent phage, the abundance of Escherichia coli K-12 (tetR) and Pseudomonas aeruginosa PAO1 (ampR + fosR) and their corresponding ARGs (tetM, tetQ, tetW, ampC, and fosA) significantly decreased in the soil after 63 days' incubation (p < 0.05). Similar results for endophytic K-12 and PAO1, and ARGs, were also obtained in lettuce tissues following combined treatment. Additionally, high throughput sequencing revealed that biochar and polyvalent phage synergetically improved the structural diversity and functional stability of the indigenous bacterial communities in soil and the endophytic ones in lettuce. Hence, this work proposes a novel biotechnology that combines biochar amendment and polyvalent phage therapy to achieve targeted inactivation of ARPB, which stimulates ARG dissipation in soil-lettuce systems.

This study focused on the role of bioaccessibility in the phenanthrene (PHE) biodegradation in diffusely contaminated soil, by combining chemical and microbiological approaches. First, we determined PHE dissipation rates and PHE sorption/desorption isotherms for two soils (PPY and Pv) presenting similar chronic PAH contamination, but different physico-chemical properties. Our results revealed that the PHE dissipation rate was significantly higher in the Pv soil compared to the PPY soil, while PHE sorption/desorption isotherms were similar. Interestingly, increases of PHE desorption and potentially of PHE bioaccessibility were observed for both soils when adding rhamnolipids (biosurfactants produced by Pseudomonas aeruginosa). Second, using 13C-PHE incubated in the same soils, we analyzed the PHE degrading bacterial communities. The combination of stable isotope probing (DNA-SIP) and 16S rRNA gene pyrosequencing revealed that Betaproteobacteria were the main PHE degraders in the Pv soil, while a higher bacterial diversity (Alpha-, Beta-, Gammaproteobacteria and Actinobacteria) was involved in PHE degradation in the PPY soil. The amendment of biosurfactants commonly used in biostimulation methods (i.e. rhamnolipids) to the two soils clearly modified the PHE sorption/desorption isotherms, but had no significant impact on PHE degradation rates and PHE-degraders identity. These results demonstrated that increasing the bioaccessibility of PHE has a low impact on its degradation and on the functional populations involved in this degradation.

Crude oil contaminant is one of the major problem to environment and its removal process considered as most challenging tool currently across the world. In this degradation study, crude oil hydrocarbons are degraded on various pH optimization conditions (pH 2, 4,6,7,8 and 10) by using two biosurfactant producing bacterial strains Pseudomonas aeruginosa PP3 and Pseudomonas aeruginosa PP4. During crude oil biodegradation, degradative enzymes alkane hydroxylase and alcohol dehydrogenase were examined and found to be higher in PP4 than PP3. Biodegradation efficiency (BE) of crude oil by both PP3 and PP4 were analysed by gas chromatography mass spectroscopy (GCMS). Based on strain PP3, the highest BE was observed in pH 2 and pH 4 were found to be 62% and 69% than pH 6, 7, 8 and 10 (47%, 47%, 49% and 45%). It reveals that PP3 was survived effectively in acidic condition and utilized the crude oil hydrocarbons. In contrast, the highest BE of PP4 was observed in pH 7 (78%) than pH4 (68%) and pH's 2, 6, 8 and 10 (52%, 52%, 43% and 53%) respectively. FTIR spectra results revealed that the presence of different functional group of hydrocarbons (OH, –CH₃, CO, C–H) in crude oil. GCMS results confirmed that both strains PP3 and PP4 were survived in acidic condition and utilized the crude oil hydrocarbons as sole carbon sources. This is the first observation on biodegradation of crude oil by the novel strains of Pseudomonas aeruginosa in acidic condition with higher BE. Overall, the extracellular enzymes and surface active compounds (biosurfactant) produced by bacterial strains were played a key role in crude oil biodegradation process.

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.