DC Electric Fields Promote Biodegradation of Waterborne Naphthalene in Biofilter Systems.

Biofiltration is a simple and low-cost method for the cleanup of contaminated water. However, the reduced availability of dissolved chemicals to surface-attached degrader bacteria may limit its efficient use at certain hydraulic loadings. When a direct current (DC) electric field is applied to an immersed packed bed, it invokes electrokinetic processes, such as electroosmotic water flow (EOF). EOF is a surface-charge-induced plug-flow-shaped movement of pore fluids. It acts at a nanometer distance above surfaces and allows the change of microscale pressure-driven flow profiles and, hence, the availability of dissolved contaminants to microbial degraders. In laboratory percolation columns, we assessed the effects of a weak DC electric field (E = 0.5 V·cm-1) on the biodegradation of waterborne naphthalene (NAH) by surface-attached Pseudomonas fluorescens LP6a. To vary NAH bioavailability, we used different NAH concentrations (C0 = 2.7, 5.1, or 7.8 × 10-5 mol·L-1) and Darcy velocities typical for biofiltration (U¯ = 0.2-1.2 × 10-4 m·s-1). In DC-free controls, we observed higher specific degradation rates (qc) at higher NAH concentrations. The qc depended on U¯, suggesting bioavailability restrictions depending on the hydraulic residence times. DC fields consistently increased qc and resulted in linearly increasing benefits up to 55% with rising hydraulic loadings relative to controls. We explain these biodegradation benefits by EOF-altered microscale flow profiles allowing for better NAH provision to bacteria attached to the collectors even though the EOF was calculated to be 100-800 times smaller than bulk water flow. Our data suggest that electrokinetic approaches may give rise to future technical applications that allow regulating biodegradation, for example, in response to fluctuating hydraulic loadings.

Nanocrystalline Cellulose to Reduce Superplasticizer Demand in 3D Printing of Cementitious Materials.

One challenge for 3D printing is that the mortar must flow easily through the printer nozzle, and after printing, it must develop compressive strength fast and high enough to support the layers on it. This requires an exact and difficult control of the superplasticizer (SP) dosing. Nanocrystalline cellulose (CNC) has gained significant interest as a rheological modifier of mortar by interacting with the various cement components. This research studied the potential of nanocrystalline cellulose (CNC) as a mortar aid for 3D printing and its interactions with SPs. Interactions of a CNC and SP with cement suspensions were investigated by means of monitoring the effect on cement dispersion (by monitoring the particle chord length distributions in real time) and their impact on mortar mechanical properties. Although cement dispersion was increased by both CNC and SP, only CNC prevented cement agglomeration when shearing was reduced. Furthermore, combining SP and CNC led to faster development of compressive strength and increased compressive strength up to 30% compared to mortar that had undergone a one-day curing process.

Comparing anticoagulant rodenticide exposure in barn owl (Tyto alba) and common kestrel (Falco tinnunculus): A biomonitoring study in an agricultural region of southeastern Spain.

Second-generation anticoagulant rodenticides (SGARs) are commonly used for rodent control, affecting various non-target wildlife species. Here, blood samples from common kestrels (Falco tinnunculus, n = 70 chicks) and barn owls (Tyto alba, n = 54 chicks and 12 adults) from Southeastern Spain were analysed using HPLC-TQ. SGAR prevalence was 68.6% in kestrel chicks, 50% in barn owl chicks and 100% in adult barn owls, with multiple SGARs in both species. Prothrombin time analysis in barn owls revealed a positive correlation with blood ΣSGARs, suggesting a potential adverse effect on coagulation. Analysis of variables potentially influencing SGAR prevalence indicated that, for kestrels, it was only related to the extent of artificial surface, showing no differences across study sites. In owlets, the highest prevalence occurred in the most urbanized study site, with human population density being a key factor. This study highlights species-specific differences in SGAR exposure, likely influenced by ecological traits. Barn owls probably encounter contaminated prey near anthropized areas, with widespread SGAR use and higher presence of target rodents. Conversely, kestrels, hunting a variety of prey often near human settlements, face consistently elevated exposure from multiple sources. Understanding these variations is crucial for effective conservation and minimizing SGAR impact on non-target wildlife.

Taxis-enhanced mineralization and co-metabolism of PAHs by bacteria in micrometer-scale environments.

Polycyclic aromatic hydrocarbons (PAHs) are associated with micropores in sediments and soils. This limits the bioaccessibility of these compounds via existing bioremediation technologies, as biodegradation is strongly influenced by the ability of bacteria to access different sizes of pores. In this work, we employed naphthalene and pyrene as model contaminants to evaluate the transformation capacity of the soil bacterium Pseudomonas putida G7 (2 × 1 µm) via mineralization and co-metabolic activity, respectively. Under non-growing conditions and in the absence of hydraulic flow, we examined how the tactic behavior of this motile bacterium influenced biodegradation of these two PAHs when passing through membranes with micrometer-sized pores (3 and 5 µm). The bacteria were spontaneously retained by the membranes, which blocked the contaminants away from a passive dosing source. However, the cells were mobilized through 5 µm pores after the application plant root exudate components (γ-aminobutyric acid, citrate and fructose) as strong chemoeffectors, which enhanced the mineralization of naphthalene and co-metabolism of pyrene. The tactic-mediated biodegradation enhancement did not occur through 3 µm pores, possibly due a physical constrain to the gradient sensing mechanism. Our results suggest that bacterial transport by chemotaxis may enhance the biotransformation of poorly bioaccessible contaminants present in micro-meter scale environments.

Single-Molecule Localization Microscopy and Tracking with a Fluorescent Mechanosensitive Probe.

A milestone in optical imaging of mechanical forces in cells has been the development of the family of flipper fluorescent probes able to report membrane tension noninvasively in living cells through their fluorescence lifetime. The specifically designed Flipper-CF3 probe with an engineered inherent blinking mechanism was recently introduced for super-resolution fluorescence microscopy of lipid ordered membranes but was too dim to be detected in lipid disordered membranes at the single-molecule level (García-Calvo, J. J. Am. Chem. Soc. 2020, 142(28), 12034-12038). We show here that the original and commercially available probe Flipper-TR is compatible with single-molecule based super-resolution imaging and resolves both liquid ordered and liquid disordered membranes of giant unilamellar vesicles below the diffraction limit. Single probe molecules were additionally tracked in lipid bilayers, enabling to distinguish membranes of varying composition from the diffusion coefficient of the probe. Differences in brightness between Flipper-CF3 and Flipper-TR originate in their steady-state absorption and fluorescence properties. The general compatibility of the Flipper-TR scaffold with single-molecule detection is further shown in super-resolution experiments with targetable Flipper-TR derivatives.

Innovative application of Nile Red (NR)-based dye for direct detection of micro and nanoplastics (MNPs) in diverse aquatic environments.

This paper presents the results of a research aimed at establishing a novel method for the detection of primary and secondary micro- and nanoplastics (MNPs), by using the fluorescence properties of the dye Nile Red-n-heptane (NR-H). The method has been applied to the detection of laboratory degraded polymers (Polystyrene, PS and Polyethylene Terephthalate, PET) as well as traceable latex microspheres in aqueous environments, showing a remarkable detection capacity and avoiding the prior extraction or processing of MNPs in natural samples, with significant time savings compared to conventional methods. The study has been carried out on various types of water, including samples from wastewater treatment plants, boreholes, seawater and synthesized seawater. The effectiveness of the staining process was evaluated by scanning electron microscopy (SEM), dynamic light scattering (DLS) and optical microscopy. As a result, a novel standardizable protocol for the rapid detection of MNPs has been established, with the potential to improve environmental protection through fast in-situ detection and identification of plastic contaminants. The limitations of the protocol in the quantification of MNPs have also been identified and further studies are proposed to overcome these limitations.

NFκB and NLRP3/NLRC4 inflammasomes regulate differentiation, activation and functional properties of monocytes in response to distinct SARS-CoV-2 proteins.

Increased recruitment of transitional and non-classical monocytes in the lung during SARS-CoV-2 infection is associated with COVID-19 severity. However, whether specific innate sensors mediate the activation or differentiation of monocytes in response to different SARS-CoV-2 proteins remain poorly characterized. Here, we show that SARS-CoV-2 Spike 1 but not nucleoprotein induce differentiation of monocytes into transitional or non-classical subsets from both peripheral blood and COVID-19 bronchoalveolar lavage samples in a NFκB-dependent manner, but this process does not require inflammasome activation. However, NLRP3 and NLRC4 differentially regulated CD86 expression in monocytes in response to Spike 1 and Nucleoprotein, respectively. Moreover, monocytes exposed to Spike 1 induce significantly higher proportions of Th1 and Th17 CD4 + T cells. In contrast, monocytes exposed to Nucleoprotein reduce the degranulation of CD8 + T cells from severe COVID-19 patients. Our study provides insights in the differential impact of innate sensors in regulating monocytes in response to different SARS-CoV-2 proteins, which might be useful to better understand COVID-19 immunopathology and identify therapeutic targets.

Percutaneous Left Atrial Appendage Closure in Patients with Non-Valvular Atrial Fibrillation and End-Stage Renal Disease on Hemodialysis: A Case Series.

Non-valvular atrial fibrillation (NVAF) is the most common cardiac arrhythmia in the general population, and its prevalence increases among patients with chronic kidney disease (CKD) undergoing hemodialysis. This population presents high risk of both hemorrhagic and thrombotic events, with little evidence regarding the use of oral anticoagulation treatment (OAT) and multiple complications arising from it; however, stroke prevention with percutaneous left atrial appendage closure (LAAC) is an alternative to be considered. We retrospectively describe the safety and efficacy of percutaneous LAAC in eight patients with NVAF and CKD on hemodialysis during a 12-month follow-up. The mean age was 78.8 years (range 64-86; SD ± 6.7), and seven patients were male. The mean CHA2DS2-VASC and HAS-BLED scores were high, 4.8 (SD ± 1.5) and 3.8 (SD ± 1.3), respectively. Seventy-five percent of the patients were referred for this intervention due to a history of major bleeding, with gastrointestinal bleeding being the most common type, while the remaining twenty-five percent of the patients were referred because of a high risk of bleeding. The percutaneous LAAC procedure was successfully completed in 100% of the patients, with complete exclusion of the appendage without complications or leaks exceeding 5 mm. There was one death not related to the procedure four days after the intervention. Among the other seven patients, no deaths, cardioembolic events or major bleeding were reported during the follow-up period. In our sample, percutaneous LAAC appears to be a safe and effective alternative to anticoagulation in patients with NVAF and CKD on hemodialysis.

Active monitoring of long-eared owl (Asio otus) nestlings reveals widespread exposure to anticoagulant rodenticides across different agricultural landscapes.

The widespread use of anticoagulant rodenticides (ARs) poses a worldwide threat to farmland wildlife. These compounds accumulate in tissues of both target and non-target species, potentially endangering both direct consumers and their predators. However, investigations on ARs in blood of free-ranging predatory birds are rare. Here, the long-eared owl (Asio otus) has been used as a model predator to assess AR exposure in different agricultural landscapes from a Mediterranean semiarid region. A total of 69 owlets from 38 nests were blood-sampled over 2021 and 2022, aiming to detect AR residues and explore factors that determine their exposure, such as land uses. In addition, prothrombin time (PT) test was conducted to assess potential effects of AR contamination. Overall, nearly all the samples (98.6 %) tested positive for at least one compound and multiple ARs were found in most of the individuals (82.6 %). Among the ARs detected, flocoumafen was the most common compound (88.4 % of the samples). AR total concentration (ΣARs) in blood ranged from 0.06 to 34.18 ng mL-1, detecting the highest levels in the most intensively cultivated area. The analysis of owl pellets from 19 breeding territories showed relevant among-site differences in the contribution of rodents and birds into the diet of long-eared owls, supporting its high dietary plasticity and indicating AR presence at multiple trophic levels. Moreover, a positive and significant correlation was found between ΣARs and PT (Rho = 0.547, p < 0.001), which demonstrates the direct effect of ARs on free-living nestlings. Our results provide a preliminary overview of AR exposure in a little-studied owl species inhabiting agricultural and rural landscapes. Despite the low detected levels, these findings indicate widespread exposure -often to multiple compounds- from early life stages, which raises concern and draws attention to an ongoing and unresolved contamination issue.

Subphthalocyanine-flipper dyads for selective membrane staining.

The design, synthesis and evaluation of a subphthalocyanine-flipper (SubPc-Flipper) amphiphilic dyad is reported. This dyad combines two fluorophores that function in the visible region (420-800 nm) for the simultaneous sensing of both ordered and disordered lipidic membranes. The flipper probes part of the dyad possesses mechanosensitivity, long fluorescence lifetimes (τ = 3.5-5 ns) and selective staining of ordered membranes. On the other hand, subphthalocyanines (SubPc) are short-lifetime (τ = 1-2.5 ns) fluorophores that are insensitive to membrane tension. As a result of a Förster Resonance Energy Transfer (FRET) process, the dyad not only retains the mechanosensitivity of flippers but also demonstrates high selectivity and emission in different kinds of lipidic membranes. The dyad exhibits high emission and sensitivity to membrane tension (Δτ = 3.5 ns) when tested in giant unilamellar vesicles (GUVs) with different membrane orders. Overall, the results of this study represent a significant advancement in the applications of flippers and dyads in mechanobiology.

Chronic liver disease-associated severe thrombocytopenia in Spain: Results from a retrospective study using machine learning and natural language processing.

BACKGROUND: Patients with chronic liver disease (CLD) often develop thrombocytopenia (TCP) as a complication. Severe TCP (platelet count<50×109/L) can increase morbidity and complicate CLD management, increasing bleeding risk during invasive procedures. OBJECTIVES: To describe the real-world scenario of CLD-associated severe TCP patients' clinical characteristics. To evaluate the association between invasive procedures, prophylactic treatments, and bleeding events in this group of patients. To describe their need of medical resource use in Spain. METHODS: This is a retrospective, multicenter study including patients who had confirmed diagnosis of CLD and severe TCP in four hospitals within the Spanish National Healthcare Network from January 2014 to December 2018. We analyzed the free-text information from Electronic Health Records (EHRs) of patients using Natural Language Processing (NLP), machine learning techniques, and SNOMED-CT terminology. Demographics, comorbidities, analytical parameters and characteristics of CLD were extracted at baseline and need for invasive procedures, prophylactic treatments, bleeding events and medical resources used in the follow up period. Frequency tables were generated for categorical variables, whereas continuous variables were described in summary tables as mean (SD) and median (Q1-Q3). RESULTS: Out of 1,765,675 patients, 1787 had CLD and severe TCP; 65.2% were male with a mean age of 54.7 years old. Cirrhosis was detected in 46% (n=820) of patients and 9.1% (n=163) had hepatocellular carcinoma. Invasive procedures were needed in 85.6% of patients during the follow up period. Patients undergoing procedures compared to those patients without invasive procedures presented higher rates of bleeding events (33% vs 8%, p<0.0001) and higher number of bleedings. While prophylactic platelet transfusions were given to 25.6% of patients undergoing procedures, TPO receptor agonist use was only detected in 3.1% of them. Most patients (60.9%) required at least one hospital admission during the follow up and 14.4% of admissions were due to bleeding events with a hospital length of stay of 6 (3, 9) days. CONCLUSIONS: NLP and machine learning are useful tools to describe real-world data in patients with CLD and severe TCP in Spain. Bleeding events are frequent in those patients who need invasive procedures, even receiving platelet transfusions as a prophylactic treatment, increasing the further use of medical resources. Because that, new prophylactic treatments that are not yet generalized, are needed.

Intended delivery mode and neonatal outcomes in pregnancies with fetal growth restriction.

Objective: To compare neonatal outcomes in pregnancies with fetal growth restriction (FGR) by intended delivery mode.Methods: This is a retrospective cohort study of singleton pregnancies with FGR that were delivered ≥34.0 weeks gestation. Neonatal outcomes were compared according to the intended delivery mode, which the attending obstetrician determined. Of note, none of the subjects had a contraindication to labor. Crude and adjusted odds ratios (ORs) and corresponding confidence intervals (CIs) were calculated via logistic regression models to assess the potential association between intended delivery mode and neonatal morbidity defined as a composite outcome (i.e. umbilical artery pH ≤7.1, 5-min Apgar score ≤7, admission to the neonatal intensive care unit, hypoglycemia, intrapartum fetal distress requiring expedited delivery, and perinatal death). A sensitivity analysis excluded intrapartum fetal distress requiring emergency cesarean delivery from the composite outcome since only patients with spontaneous labor or labor induction could meet this criterion. Potential confounders in the adjusted effects models included maternal age, body mass index, hypertensive disorders, diabetes, FGR type (i.e. early or late), and oligohydramnios.Results: Seventy-two (34%) patients had an elective cesarean delivery, 73 (34%) had spontaneous labor and were expected to deliver vaginally, and 67 (32%) underwent labor induction. The composite outcome was observed in 65.3%, 89%, and 88.1% of the groups mentioned above, respectively (p < 0.001). Among patients with spontaneous labor and those scheduled for labor induction, 63% and 47.8% required an emergency cesarean delivery for intrapartum fetal distress. Compared to elective cesarean delivery, spontaneous labor (OR 4.32 [95% CI 1.79, 10.42], p = 0.001; aOR 4.85 [95% CI 1.85, 12.66], p = 0.001), and labor induction (OR 3.92 [95% CI 1.62, 9.49] p = 0.002; aOR 5.29 [95% CI 2.01, 13.87], p = 0.001) had higher odds of adverse neonatal outcomes.Conclusion: In this cohort of FGR, delivering at ≥34 weeks of gestation, pregnancies with spontaneous labor, and those that underwent labor induction had higher odds of neonatal morbidity than elective cesarean delivery.

T regulatory lymphocytes specific for SARS-CoV-2 display increased functional plasticity.

The study of phenotypic and functional characteristics of immune cells involved in host response to SARS-CoV-2 is relevant for understanding COVID-19 pathogenesis and individual differences in disease progression. We have analyzed chemokine receptor expression in SARS-CoV-2-specific CD4+ T lymphocytes from vaccinated donors, and have found an increase of CCR9+ and CCR6+ cells. CCR9+ specific CD4+ cells are enriched in T regulatory (Treg) lymphocytes. These cells specifically show heterogeneous regulatory activity, associated with different profiles of CCR9/CCR6 expression, individual differences in IL-10 and IL-17 production, and variable FoxP3 and Notch4 expression. A higher heterogeneity in FoxP3 is selectively observed in convalescent individuals within vaccinated population. Accordingly, SARS-CoV-2-specific CD4+ lymphocytes from COVID-19 patients are also enriched in CCR9+ and CCR6+ cells. CCR6+ specific Treg lymphocytes are mainly increased in critically ill individuals, indicating a preferential role for these cells in lung injury pathogenesis. We provide experimental evidence for a SARS-CoV-2-specific Treg population with increased plasticity, which may contribute to the differential pathogenic response against SARS-CoV-2 among individuals, and underlie the development of autoimmune conditions following SARS-CoV-2 infection.

Development of Lightweight Mortars Using Sustainable Low-Density Glass Aggregates from Secondary Raw Materials.

In this study, different lightweight expanded glass aggregates (LEGAs) were produced from glass cullet and various carbonated wastes, through a thermal impact process. The effects of LEGA microstructure and morphology on both the adherence to the cement paste and the mechanical properties of mortars after 28 days of curing were studied. The properties of lightweight mortars made of either LEGAs or expanded clay aggregates were compared. The results demonstrated the feasibility of using LEGAs to produce glass lightweight aggregate mortar, with flexural and compressive strength values ranging from 5.5 to 8.2 MPa and from 28.1 to 47.6 MPa, respectively. The differences in mechanical properties were explained according to the microstructures of the fracture surfaces. Thus, arlite-type ceramic aggregates presented surface porosities that allowed mortar intrusion and the formation of an interconnected interface; although the surfaces of the vitreous aggregates were free from porosity due to their vitreous nature, the mortars obtained from different wastes presented compressive and flexural strengths in the range of lightweight mortars.

State of the Art Membrane Science and Technology in the Iberian Peninsula 2021-2022.

This Special Issue of the journal Membranes arises from the need to highlight the developments in the field of membrane research and membrane processes that have been emerging in recent years by researchers and research groups based in the Iberian Peninsula [...].

Automated Liquid-Liquid Displacement Porometry (LLDP) for the Non-Destructive Characterization of Ultrapure Water Purification Filtration Devices.

This scientific publication presents a novel modification of the liquid-liquid displacement porosimetry (LLDP) method, aiming for the non-destructive automated analysis of water purification membrane filtration devices in the microfiltration (MF) and ultrafiltration (UF) range. The technical adaptation of LLDP enables the direct in-line porosimetric analysis of commercial filtration devices, avoiding the filtration devices' destruction. Six commercially available filtration devices with polyethersulfone (PES) and polysulfone (PS) membranes were studied using an improved device developed by the IFTS, which was based on a commercial LLDP instrument. The filtration devices were evaluated in three different configurations: flat disks, hollow fibers, and pleated membranes. The results obtained using the proposed method were compared with other characterization techniques, including submicronic efficiency retention, image analysis of scanning electron microscopy (SEM), and gas-liquid displacement porosimetry (GLDP). The comparison of the results demonstrated that the proposed method accurately determined the porosimetric characteristics of the filters. It proved to be a precise technique for the non-destructive in-line evaluation of filter performance, as well as for periodic quality control and the fouling degree assessment of commercial filtration devices. This modified LLDP approach offers significant potential for the advanced characterization and quality assessment of water purification membrane filtration devices, contributing to improved understanding and optimization of their performance.

Effect of pH Hydrolysis on the Recovery of Antimony from Spent Electrolytes from Copper Production.

This study examined how pH hydrolysis affects the recovery process for antimony extracted from spent electrolytes. Various OH- reagents were used to adjust the pH levels. The findings reveal that pH plays a crucial role in determining the optimal conditions for extracting antimony. The results show that NH4OH and NaOH are more effective compared to water, with optimal conditions at pH 0.5 for water and pH 1 for NH4OH and NaOH, resulting in average antimony extraction yields of 90.4%, 96.1%, and 96.7%, respectively. Furthermore, this approach helps to improve both crystallography and purity related to recovered antimony samples obtained through recycling processes. The solid precipitates obtained lack a crystalline structure, making it difficult to identify the compounds formed, but element concentrations suggest the presence of oxychloride or oxide compounds. Arsenic is incorporated into all solids, affecting the purity of the product, and water showing higher antimony content (68.38%) and lower arsenic values (8%) compared to NaOH and NH4OH. Bismuth integration into solids is less than arsenic (less than 2%) and remains unaffected by pH levels except in tests with water, where a bismuth hydrolysis product is identified at pH 1, accounting for the observed reduction in antimony extraction yields.

Silica Nanoparticle/Fluorescent Dye Assembly Capable of Ultrasensitively Detecting Airborne Triacetone Triperoxide: Proof-of-Concept Detection of Improvised Explosive Devices in the Workroom.

We describe the proof of concept of a portable testing setup for the detection of triacetone triperoxide (TATP), a common component in improvised explosive devices. The system allows for field testing and generation of real-time results to test for TATP vapor traces in air by simply using circulation of the air samples through the sensing mechanism under the air conditioning system of an ordinary room. In this way, the controlled trapping of the analyte in the chemical sensor gives reliable results at extremely low concentrations of TATP in air under real-life conditions, suitable for daily use in luggage storage for airlines or a locker room for a major sporting event. The reported fluorescent methodology is very sensitive and selective, allowing for the trapping of triacetone triperoxide in the chemical sensor to give reliable results at very low concentrations in air under ambient conditions, by comparing the fluorescence of the material before and after exposition to TATP traces in air.

Risk reductions during pyrene biotransformation and mobilization in a model plant-bacteria-biochar system.

The productive application of motile microorganisms for degrading hydrophobic contaminants in soil is one of the most promising processes in modern remediation due to its sustainability and low cost. However, the incomplete biodegradation of the contaminants and the formation of the intermediary metabolites in the process may increase the toxicity in soil during bioremediation, and motile inoculants may mobilize the pollutants through biosorption. Therefore, controlling these factors should be a fundamental part of soil remediation approaches. The aim of this study was to evaluate the sources of risk associated with the cometabolism-based transformation of 14C-labeled pyrene by inoculated Pseudomonas putida G7 and identify ways to minimize risk. Our model scenario examined the increase in bioaccessibility to a distant source of contamination facilitated by sunflower (Helianthus annuus L.) roots. A biochar trap for mobilized pollutant metabolites and bacteria has also been employed. The experimental design consisted of pots filled with a layer of sand with 14C-labeled pyrene (88 mg kg-1) as a contamination focus located several centimeters from the inoculation point. Half of the pots included a biochar layer at the bottom. The pots were incubated in a greenhouse with sunflower plants and P. putida G7 bacteria. Pots with sunflower plants showed a higher biodegradation of pyrene, its mobilization as metabolites through the percolate and the roots, and bacterial mobilization toward the source of contamination, also resulting in increased pyrene transformation. In addition, the biochar layer efficiently reduced the concentrations of pyrene metabolites collected in the leachates. Therefore, the combination of plants, motile bacteria and biochar safely reduced the risk caused by the biological transformation of pyrene.