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The hexachlorocyclohexane isomers (HCH) are long-banned pesticides. Even though their use has been prohibited for decades, their presence in the environment is still reported worldwide. Wetland + is a registered trademark of the remedial treatment technology consisting of an aerobic sedimentary tank, a permeable reactive barrier, a biosorption system, and an aerobic wetland. This proven method combines a reductive treatment known from PRBs with the natural wetland self-cleaning processes. The average efficiency of the system is 96.8% for chlorobenzenes (ClB) and 81.7% for HCH, during the first 12 months of the system operation. The presence of the genes encoding enzymes involved in the degradation of the HCH compounds indicates that the removal of HCH and ClB occurs not only by chemical removal but also through aerobic and anaerobic combining biodegradation. Changes in abundance and the composition of the diatom community were found to be suitable indicators of the water quality and of the impact of the Wetland + operation on the water ecosystem. The system's annual operation exhibited a markedly higher number of diatom species in the closing profiles of the Ostrovský Creek, the Wetland + effluent recipient.
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Ecossistema , Hexaclorocicloexano , Hexaclorocicloexano/química , Hexaclorocicloexano/metabolismo , Áreas Alagadas , Biodegradação Ambiental , BiodiversidadeRESUMO
Halogenated organic compounds are naturally occurring in subsurface environments; however, accumulation of the degradative intermediate cis-1,2-dichloroethene (cDCE) at soil and groundwater sites contaminated with xenobiotic chlorinated ethenes is a global environmental and public health issue. Identifying microorganisms capable of cDCE degradation in these environments is of interest because of their potential application to bioremediation techniques. In this study, we sequenced, assembled, and analyzed the complete genome of Acinetobacter pittii CEP14, a strain isolated from chloroethene-contaminated groundwater, that has demonstrated the ability for aerobic cometabolic degradation of cDCE in the presence of n-hexane, phenol, and toluene. The A. pittii CEP14 genome consists of a 3.93 Mbp-long chromosome (GenBank accession no. CP084921) with a GC content of 38.9% and three plasmids (GenBank accession no. CP084922, CP084923, and CP084924). Gene function was assigned to 83.4% of the 3,930 coding DNA sequences. Functional annotation of the genome revealed that the CEP14 strain possessed all genetic elements to mediate the degradation of a range of aliphatic and aromatic compounds, including n-hexane and phenol. In addition, it harbors gene clusters involved in cytosol detoxification and oxidative stress resistance, which could play a role in the mitigation of toxic chemical intermediates that can arise during the degradation of cDCE. Gene clusters for heavy metal and antibiotic resistance were also identified in the genome of CEP14. These results suggest that CEP14 may be a versatile degrader of xenobiotic compounds and well-adapted to polluted environments, where a combination of heavy metal and organic compound pollution is often found.
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Fenóis , Xenobióticos , Acinetobacter , Biodegradação Ambiental , Dicloroetilenos , GenômicaRESUMO
The application of zero-valent iron particles (ZVI) for the treatment of heavily polluted environment and its biological effects have been studied for at least two decades. Still, information on the impact on bacterial metabolic pathways is lacking. This study describes the effect of microscale and nanoscale ZVI (mZVI and nZVI) on the abundance of different metabolic pathways in freshwater bacterial communities. The metabolic pathways were inferred from metabolism modelling based on 16S rRNA gene sequence data using paprica pipeline. The nZVI changed the abundance of numerous metabolic pathways compared to a less influencing mZVI. We identified the 50 most affected pathways, where 31 were related to degradation, 17 to biosynthesis, and 2 to detoxification. The linkage between pathways was two times higher in nZVI samples compared to mZVI, and was specifically higher considering the arsenate detoxification II pathway. Limnohabitans and Roseiflexus were linked to the same pathways in both nZVI and mZVI. The prediction of metabolic pathways increases our knowledge of the impacts of nZVI and mZVI on freshwater bacterioplankton.
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Chloroflexi , Poluentes Químicos da Água , Água Doce , Genes de RNAr , Ferro , RNA Ribossômico 16S/genéticaRESUMO
The second generation energy crop Miscanthus x giganteus (Mxg) was cultivated in pots with mixtures of clean and petroleum industry contaminated soil affected by petroleum, Pb, Zn contamination and high salinity. The survival rate reached 100%, nevertheless the biomass parameters were negatively affected even in the lowest proportion of contaminated soil. In the lowest contamination, where the plant grew still quite successfully, C10-C40 degradation was significantly enhanced compared to the unplanted control with degradation of 58 ± 14%. The plant contribution to aliphatics degradation was significantly correlated with biomass, thus it was negligible in higher contamination. A similar pattern was documented in development of the soil bacterial community. The shift in community structure after Mxg cultivation was observed mainly in the soil with the lowest contaminant proportion, though an increase of bacterial diversity in the miscanthus rhizosphere was observed in all cases. Relative abundance of Actinobacteria was reduced on behalf of several less abundant phyla (Verrucomicrobia, Bacterioides, Acidobacteria). The majority of genera identified as potential petroleum degraders (Pseudomonas, Shinella, Altererythrobacter, Azospirillum, Mesorhizobium, Dyella) were more abundant in contaminated soil with miscanthus, suggesting that Mxg could be a promising crop for phytomanagement of petroleum contaminated soils but salt phytotoxicity needs to be mitigated first.
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As nanoremediation strategies for in-situ groundwater treatment extend beyond nanoiron-based applications to adsorption and oxidation, ecotoxicological evaluations of newly developed materials are required. The biological effects of four new materials with different iron (Fe) speciations ([i] FerMEG12 - pristine flake-like milled Fe(0) nanoparticles (nZVI), [ii] Carbo-Iron® - Fe(0)-nanoclusters containing activated carbon (AC) composite, [iii] Trap-Ox® Fe-BEA35 (Fe-zeolite) - Fe-doped zeolite, and [iv] Nano-Goethite - 'pure' FeOOH) were studied using the unicellular green alga Chlamydomonas sp. as a model test system. Algal growth rate, chlorophyll fluorescence, efficiency of photosystem II, membrane integrity and reactive oxygen species (ROS) generation were assessed following exposure to 10, 50 and 500â¯mgâ¯L-1 of the particles for 2â¯h and 24â¯h. The particles had a concentration-, material- and time-dependent effect on Chlamydomonas sp., with increased algal growth rate after 24â¯h. Conversely, significant intracellular ROS levels were detected after 2â¯h, with much lower levels after 24â¯h. All Fe-nanomaterials displayed similar Z-average sizes and zeta-potentials at 2â¯h and 24â¯h. Effects on Chlamydomonas sp. decreased in the order FerMEG12 > Carbo-Iron® > Fe-zeolite > Nano-Goethite. Ecotoxicological studies were challenged due to some particle properties, i.e. dark colour, effect of constituents and a tendency to agglomerate, especially at high concentrations. All particles exhibited potential to induce significant toxicity at high concentrations (500â¯mgâ¯L-1), though such concentrations would rapidly decrease to mg or µgâ¯L-1 in aquatic environments, levels harmless to Chlamydomonas sp. The presented findings contribute to the practical usage of particle-based nanoremediation in environmental restoration.
Assuntos
Chlamydomonas/efeitos dos fármacos , Recuperação e Remediação Ambiental/métodos , Ferro/farmacologia , Nanoestruturas/química , Adsorção , Membrana Celular/efeitos dos fármacos , Carvão Vegetal/química , Chlamydomonas/crescimento & desenvolvimento , Chlamydomonas/metabolismo , Água Subterrânea , Ferro/química , Compostos de Ferro/química , Minerais/química , Oxirredução , Espécies Reativas de Oxigênio/metabolismo , Zeolitas/químicaRESUMO
Although the pesticide hexachlorocyclohexane (HCH) and its isomers have long been banned, their presence in the environment is still reported worldwide. In this study, we investigated the bioaccumulation potential of α, ß, and δ hexachlorocyclohexane (HCH) isomers in black alder saplings (Alnus glutinosa) to assess their environmental impact. Each isomer, at a concentration of 50 mg/kg, was individually mixed with soil, and triplicate setups, including a control without HCH, were monitored for three months with access to water. Gas chromatography-mass spectrometry revealed the highest concentrations of HCH isomers in roots, decreasing towards branches and leaves, with δ-HCH exhibiting the highest uptake (roots-14.7 µg/g, trunk-7.2 µg/g, branches-1.53 µg/g, leaves-1.88 µg/g). Interestingly, α-HCH was detected in high concentrations in ß-HCH polluted soil. Phytohormone analysis indicated altered cytokinin, jasmonate, abscisate, and gibberellin levels in A. glutinosa in response to HCH contamination. In addition, amplicon 16S rRNA sequencing was used to study the rhizosphere and soil microbial community. While rhizosphere microbial populations were generally similar in all HCH isomer samples, Pseudomonas spp. decreased across all HCH-amended samples, and Tomentella dominated in ß-HCH and control rhizosphere samples but was lowest in δ-HCH samples.
Assuntos
Alnus , Poluentes do Solo , Hexaclorocicloexano/análise , Biodegradação Ambiental , RNA Ribossômico 16S/genética , Poluentes do Solo/análise , SoloRESUMO
Nanoplastics are anticipated to be ubiquitous in various environmental compartments. However, challenges in analytical methods hinder our understanding of risks related to specific nanplastics characteristics such as size and chemical compositions, and interactions between nanoplastics and microorganisms. In this study, we applied fit-for-purpose analytical methods and techniques to understand how nanoplastic chemical composition influences their interaction with bacteria collected from activated sludge. When exposed to polystyrene (PS) and polyvinyl chloride (PVC) nanoplastics for 5 days, the nanoplastics attached to the bacteria. Specifically, on day 1, there was a significant predominance of PS nanoplastics over PVC ones of similar size and shape, possibly due to differences in their chemical composition. After 5 days, there is a substantial decrease in nanoplastics attached to bacteria, suggesting bacterial defence mechanisms may reduce particles attachment over time. The overall bacterial community structure demonstrated a high degree of resilience. This resilience highlights the ability of microbial communities to maintain their structure despite nanoplastic stressors, as evidenced by consistent alpha diversity, PCoA, and PERMANOVA results. Understanding these mechanisms is crucial for assessing nanoplastic fate and thus environmental impacts.
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Bio-based plastics (BP) produced from renewable biomass resources, such as high-density polyethylene (HDPE), polylactic acid (PLA) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), is currently increasing in terms of both products and applications. However, their biodegradability and environmental fate are not yet fully understood, especially in freshwaters. Here, we present the results of an in-situ study in a freshwater reservoir, where we submerged HDPE, PLA and PHBV microscale BP (mBP) in dialysis bags to enable exchange of small organic and inorganic molecules, including nutrients, with the surrounding water. After one and two months, the bacterial biofilm that formed on each mBP was characterised by 16S rRNA amplicon sequencing. After two-months, Oxalobacteraceae, Pedosphaeraceae, Flavobacteriaceae (Flavobacterium) and Chitinophagaceae (Ferruginibacter) had increased by up to four times. Both these and other common members (≥1 % relative total biomass) of the microbial community were similarly abundant on all mBP. Low-abundance (0.3-1 %) bacterial taxa, however, were significantly more diverse and differed on each mBP. Notably, some low-abundance families and genera increased on specific materials, e.g. Sphingomonadaceae on HDPE, Sphingobacteriaceae on PHBV, Gemmatimonas and Crenothrix on PLA. Overall, abundant bacteria were regarded as a pioneering community, while low-abundance bacteria were more diverse and preferred mBP types in the early stages of biofilm formation on mBP. It could be influenced by the environmental conditions, where nutrient levels and low temperatures might shape the low-abundance of attached bacterial communities than the plastic material itself.
Assuntos
Microplásticos , Plásticos , Humanos , RNA Ribossômico 16S/genética , Polietileno , Diálise Renal , Água Doce , Biofilmes , Bactérias , PoliésteresRESUMO
Chlorinated ethenes (CEs) are common and persistent contaminants of soil and groundwater. Their degradation is mostly driven by a process of bacterial reductive dechlorination (also called organohalide respiration) in anaerobic conditions. This study summarizes the outcomes of the long-term in-situ application of glycerol for the enhanced reductive dechlorination of CEs on a highly contaminated site. Glycerol injection resulted in an almost immediate increase in the abundance of fermentative Firmicutes, which produce essential sources of carbon (acetate) and electrons (H2) for organohalide-respiring bacteria (OHRB) and change groundwater conditions to be suitable for OHRB growth. The decreased redox potential of groundwater promoted also the proliferation of sulfate-reducing bacteria, which compete for electron donors with OHRB but at the same time support their growth by producing essential corrinoids and acetate. A considerable increase in the abundance of OHRB Dehalococcoides, concurrently with vinyl chloride (VC) reductase gene levels, was revealed by real time polymerase chain reaction (qPCR) method. Consistent with the shifts in bacterial populations, the concentrations of pollutants tetrachloroethylene and trichloroethylene decreased during the monitoring period, with rising levels of cis-1,2-dichloroethylene, VC, and most importantly, the final CE degradation products: ethene and ethane. Our study implies the importance of syntrophic bacterial interactions for successful and complete CE degradation and evaluates glycerol as convenient substrate to enhance reductive dechlorination and as an effective source of electrons for OHRB.
Assuntos
Chloroflexi , Corrinoides , Microbiota , Tetracloroetileno , Tricloroetileno , Cloreto de Vinil , Poluentes Químicos da Água , Tetracloroetileno/metabolismo , Tricloroetileno/metabolismo , Glicerol/metabolismo , Biodegradação Ambiental , Poluentes Químicos da Água/metabolismo , Bactérias/genética , Bactérias/metabolismo , Solo , Carbono/metabolismo , Oxirredutases/metabolismo , Sulfatos/metabolismo , Chloroflexi/metabolismoRESUMO
Concrete as an important component of an engineered barrier system in deep geological repositories (DGR) for radioactive waste may come into contact with bentonite, or other clays, rich in indigenous microorganisms, with potentially harmful impacts on barrier integrity. Our study aimed to assess the effect of a concrete environment on indigenous bentonite and groundwater microbial communities as these particular conditions will select for the potentially harmful microorganisms to the concrete in the future DGR. The two-month experiment under anoxic conditions consisted of crushed, aged, low-pH concrete, Czech Ca-Mg bentonite, and anoxic groundwater, with control samples without concrete or with sterile groundwater. The microbial diversity and proliferation were estimated by qPCR and 16S rRNA gene amplicon sequencing. The presence of concrete had a strong effect on microbial diversity and reduced the increase in total microbial biomass, though low-pH concrete harbored indigenous bacteria. The growth of sulfate reducers was also limited in concrete samples. Several genera, such as Massilia, Citrifermentans, and Lacunisphaera, dominant in bentonite controls, were suppressed in concrete-containing samples. In contrast, genera such as Bacillus, Dethiobacter and Anaerosolibacter specifically proliferated in the presence of concrete. Genera such as Thermincola or Pseudomonas exhibited high versatility and proliferated well under both conditions. Because several of the detected bacterial genera are known to affect concrete integrity, further long-term studies are needed to estimate the effect of bentonite and groundwater microorganisms on concrete stability in future DGR.
Assuntos
Microbiota , Resíduos Radioativos , Bentonita , Concentração de Íons de Hidrogênio , RNA Ribossômico 16S/genética , Resíduos Radioativos/análise , SuspensõesRESUMO
Complex assessment of gradual changes in scaffold morphology and stiffness is an essential step in bone filler development. Current approach, however, does not reflect long term cell proliferation effect as the mechanical tests are usually conducted on pristine materials without cells or cell influence on material stiffness is evaluated after one time period only. Here, biocompatible silk fibroin (SF) porous scaffolds envisioned for bone defect filling were prepared by dissolving of fibroin fibers, followed by dialysis, freeze-drying and final stabilization. Particular attention was devoted to the influence of bone cell proliferation up to 2 months on the stiffness of the material. The morphology of the material was studied in terms of its inner structure and the overall changes in the surface characteristics due to proliferation of MG 63 bone cell line. The SF scaffold stiffness significantly increased during first month followed by its decline during second month due to bone cell seeding. After 2 months, the SF scaffold was completely colonized, which resulted in a gradual decay of its structure. The length of degradation due to bone cell proliferation and mechanical behavior corresponded to the requirements set for reasonable filler material indicating that porous SF scaffolds comprise a promising biomaterial for bone regeneration.
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Nanoscale zero-valent iron (nZVI) has increasingly been applied to remediate aquifers polluted by organochlorines or heavy metals. As a result, bacteria in the vicinity of remediate action can be stressed by surplus iron released from nZVI. However, the understanding of the iron stress defense pathways during this process is currently incomplete. Therefore, we aimed to elucidate the physiological and transcriptomic response of the bacterium, Pseudomonas putida NCTC 10936, to 100 mg/L of nZVI and 44.5 µg/L of dissolved iron obtained from nZVI suspension. Cell viability was neither affected by nZVI nor dissolved iron, although the dissolved iron caused stress that altered the cell physiology and caused the generation of smaller cells, whereas cells were elongated in the presence of nZVI. Transcriptomic analysis confirmed the observed stronger physiological effect caused by dissolved iron (in total 3839 differentially expressed genes [DEGs]) than by nZVI (945 DEGs). Dissolved iron (but not nZVI) activated genes involved in oxidative stress-related pathways, antioxidant activity, carbohydrate and energy metabolism, but downregulated genes associated with flagellar assembly proteins and two-component systems involved in sensing external stimuli. As a result, bacteria very effectively faced oxidative insults and cell viability was not affected.
Assuntos
Metais Pesados , Pseudomonas putida , Ferro , Oxirredução , Estresse Oxidativo , Pseudomonas putida/genéticaRESUMO
Rising concern about emerging and already persisting pollutants in water has urged the scientific community to develop novel remedial techniques. A new group of remediation methods is based on the modification of nanoscale zero-valent iron particles (nZVI), which are well known for treating volatile organic compounds and heavy metals. The properties of nZVI may be further enhanced by modifying their structure or surface using "green" polymers. Herein, nZVI was modified by a ß-cyclodextrin polymer (ß-CDP), which is considered an environmentally safe and inexpensive adsorbent of contaminants. This composite was used for the first time for the degradation of sulfamethoxazole (SMX). Coating by ß-CDP not only enhanced the degradation of SMX (>95%, under 10 min) by the nanoparticles in a wide pH range (3-9) and enabled their efficient reusability (for three cycles) but also made the coated nZVI less toxic to the model bioindicator microalga Raphidocelis subcapitata. Moreover, degradation products of SMX were found to be less toxic to Escherichia coli bacteria and R. subcapitata microalga, contrary to the SMX antibiotic itself, indicating a simple and eco-friendly cleaning process. This research aims to further stimulate and develop novel remedial techniques based on nZVI, and provides a potential application in the degradation of antibiotics in a wide pH range. Moreover, the wealth of available cyclodextrin materials used for surface modification may open a way to discover more efficient and attractive composites for environmental applications.
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Ciclodextrinas , Metais Pesados , Poluentes Químicos da Água , Celulose , Ferro/química , Sulfametoxazol/toxicidade , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidadeRESUMO
Decades of antibiotic use and misuse have generated selective pressure toward the rise of antibiotic-resistant bacteria, which now contaminate our environment and pose a major threat to humanity. According to the evolutionary "Red queen theory", developing new antimicrobial technologies is both urgent and mandatory. While new antibiotics and antibacterial technologies have been developed, most fail to penetrate the biofilm that protects bacteria against external antimicrobial attacks. Hence, new antimicrobial formulations should combine toxicity for bacteria, biofilm permeation ability, biofilm deterioration capability, and tolerability by the organism without renouncing compatibility with a sustainable, low-cost, and scalable production route as well as an acceptable ecological impact after the ineluctable release of the antibacterial compound in the environment. Here, we report on the use of silver nanoparticles (NPs) doped with magnetic elements (Co and Fe) that allow standard silver antibacterial agents to perforate bacterial biofilms through magnetophoretic migration upon the application of an external magnetic field. The method has been proved to be effective in opening micrometric channels and reducing the thicknesses of models of biofilms containing bacteria such as Enterococcus faecalis, Enterobacter cloacae, and Bacillus subtilis. Besides, the NPs increase the membrane lipid peroxidation biomarkers through the formation of reactive oxygen species in E. faecalis, E. cloacae, B. subtilis, and Pseudomonas putida colonies. The NPs are produced using a one-step, scalable, and environmentally low-cost procedure based on laser ablation in a liquid, allowing easy transfer to real-world applications. The antibacterial effectiveness of these magnetic silver NPs may be further optimized by engineering the external magnetic fields and surface conjugation with specific functionalities for biofilm disruption or bactericidal effectiveness.
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Anti-Infecciosos , Nanopartículas Metálicas , Prata/farmacologia , Biofilmes , Antibacterianos/farmacologia , Enterococcus faecalis , Fenômenos Magnéticos , Testes de Sensibilidade MicrobianaRESUMO
The organochlorine pesticides (OCPs) have raised concerns about being persistent and toxic to the environment. Phytoremediation techniques show promise for the revitalization of polluted soils. The current study focused on optimizing the phytoremediation potential of Miscanthus sinensis And. (M. sinensis), second-generation energy crop, by exploring two soil amendments: Tween 20 and activated carbon (AC). The results showed that when M. sinensis grew in OCP-polluted soil without amendments to it, the wide range of compounds, i.e., α-HCH, ß-HCH, γ-HCH, 2.4-DDD, 4.4-DDE, 4.4-DDD, 4.4-DDT, aldrin, dieldrin, and endrin, was accumulated by the plant. The introduction of soil amendments improved the growth parameters of M. sinensis. The adding of Tween 20 enhanced the absorption and transmigration to aboveground biomass for some OCPs; i.e., for γ-HCH, the increase was by 1.2, for 4.4-DDE by 8.7 times; this effect was due to the reduction of the hydrophobicity which made pesticides more bioavailable for the plant. The adding of AC reduced OCPs absorption by plants, consequently, for γ-HCH by 2.1 times, 4.4-DDD by 20.5 times, 4.4-DDE by 1.4 times, 4.4-DDT by 8 times, α-HCH was not adsorbed at all, and decreased the translocation to the aboveground biomass: for 4.4-DDD by 31 times, 4.4-DDE by 2.8 times, and γ-HCH by 2 times; this effect was due to the decrease in the bioavailability of pesticides. Overall, the amendment of OCP-polluted soil by Tween 20 speeds the remediation process, and incorporation of AC permitted to produce the relatively clean biomass for energy.
Assuntos
Hidrocarbonetos Clorados , Praguicidas , Poluentes do Solo , Biodegradação Ambiental , Carvão Vegetal , Monitoramento Ambiental , Hidrocarbonetos Clorados/análise , Praguicidas/análise , Polissorbatos , Solo , Poluentes do Solo/análiseRESUMO
Biologically active micropollutants that contain diverse phenolic/aromatic structures are regularly present in wastewater effluents and are even found in drinking water. Advanced green technologies utilizing immobilized laccase and/or peroxidase, which target these micropollutants directly, may provide a reasonable alternative to standard treatments. Nevertheless, the use of these enzymes is associated with several issues that may prevent their application, such as the low activity of laccase at neutral and basic pH or the necessity of hydrogen peroxide addition as a co-substrate for peroxidases. In this study, the activity of laccase from Trametes versicolor and horseradish peroxidase was evaluated across a range of commonly used substrates (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), syringaldazine, and guaiacol). Moreover, conditions for their optimal performance were explored along with an assessment of whether these conditions accurately reflect the effectivity of both enzymes in the degradation of a mixture of bisphenol A, 17α-ethinylestradiol, triclosan, and diclofenac in tap drinking water and secondary wastewater effluent. Laccase and horseradish peroxidase showed optimal activity at strongly acidic pH if ABTS was used as a substrate. Correspondingly, the activities of both enzymes detected using ABTS in real waters were significantly enhanced by adding approximately 2.5% (v/v) of McIlvaine's buffer. Degradation of a mixture of micropollutants in wastewater with 2.5% McIlvaine's buffer (pH 7) resulted in a substantial decrease in estrogenic activity. Low degradation efficiency of micropollutants by laccase was observed in pure McIlvaine's buffer of pH 3 and 7, compared with efficient degradation in tap water of pH 7.5 without buffer. This study clearly shows that enzyme activity needs to be evaluated on micropollutants in real waters as the assessment of optimal conditions based on commonly used substrates in pure buffer or deionized water can be misleading.
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Água Potável , Lacase , Peroxidase do Rábano Silvestre , Polyporaceae , Trametes , Águas ResiduáriasRESUMO
We examined microbial corrosion of carbon steel in synthetic bentonite pore water inoculated with natural underground water containing microorganisms over a period of 780-days under sterile and anaerobic conditions. Corrosion behaviour was determined using the mass loss method, SEM-EDS analysis and Raman spectroscopy, while qualitative and quantitative changes in the microbial community were analysed using molecular-biological tools (16S rDNA amplicon sequencing and qPCR analysis, respectively). Corrosion rates were significantly higher in the biotic environment (compared with an abiotic environment), with significant localisation of corrosion attacks of up to 1 mm arising within 12-months. Nitrate reducing bacteria, such as Pseudomonas, Brevundimonas and Methyloversatilis, dominated the microbial consortium, the high abundance of Methyloversatilis correlating with periods of highest localised corrosion penetrations, suggesting that this bacterium plays an important role in microbially influenced corrosion. Our results indicate that nitrate-reducing bacteria could represent a potential threat to waste canisters under nuclear repository conditions.
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Resíduos Radioativos , Aço , Anaerobiose , Carbono , CorrosãoRESUMO
Abiotic and biotic remediation of chlorinated ethenes (CEs) in groundwater from a real contaminated site was studied using biochar-based composites containing nanoscale zero-valent iron (nZVI/BC) and natural resident microbes/specific CE degraders supported by a whey addition. The material represented by the biochar matrix decorated by isolated iron nanoparticles or their aggregates, along with the added whey, was capable of a stepwise dechlorination of CEs. The tested materials (nZVI/BC and BC) were able to decrease the original TCE concentration by 99% in 30 days. Nevertheless, regarding the transformation products, it was clear that biotic as well as abiotic transformation mechanisms were involved in the transformation process when nonchlorinated volatiles (i.e., methane, ethane, ethene, and acetylene) were detected after the application of nZVI/BC and nZVI/BC with whey. The whey addition caused a massive increase in bacterial biomass in the groundwater samples (monitored by 16S rRNA sequencing and qPCR) that corresponded with the transformation of trichloro- and dichloro-CEs, and this process was accompanied by the formation of less chlorinated products. Moreover, the biostimulation step also eliminated the adverse effect caused by nZVI/BC (decrease in microbial biomass after nZVI/BC addition). The nZVI/BC material or its aging products, and probably together with vinyl chloride-respiring bacteria, were able to continue the further reductive dechlorination of dichlorinated CEs into nonhalogenated volatiles. Overall, the results of the present study demonstrate the potential, feasibility, and environmental safety of this nanobioremediation approach.
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Água Subterrânea , Poluentes Químicos da Água , Carvão Vegetal , RNA Ribossômico 16S/genética , SolventesRESUMO
Compacted bentonites are one of the best sealing and backfilling clays considered for use in Deep Geological Repositories of radioactive wastes. However, an in-depth understanding of their behavior after placement in the repository is required, including if the activity of indigenous microorganisms affects safety conditions. Here we provide an optimized phenol:chloroform based protocol that facilitates higher DNA-yields when other methods failed. To demonstrate the efficiency of this method, DNA was extracted from acetate-treated bentonites compacted at 1.5 and 1.7 g/cm3 densities after 24 months anoxic incubation. Among the 16S rRNA gene sequences identified, those most similar to taxa mediating biogeochemical sulfur cycling included sulfur oxidizing (e.g., Thiobacillus, and Sulfurimonas) and sulfate reducing (e.g., Desulfuromonas and Desulfosporosinus) bacteria. In addition, iron-cycling populations included iron oxidizing (e.g., Thiobacillus and Rhodobacter) plus reducing taxa (e.g., Geobacillus). Genera described for their capacity to utilize acetate as a carbon source were also detected such as Delftia and Stenotrophomonas. Lastly, microscopic analyses revealed pores and cracks that could host nanobacteria or spores. This study highlights the potential role of microbial driven biogeochemical processes in compacted bentonites and the effect of high compaction on microbial diversity in Deep Geological Repositories.
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Resíduos Radioativos , Bactérias/genética , Bentonita , Argila , RNA Ribossômico 16S/genética , Resíduos Radioativos/análiseRESUMO
Nanoscale zero-valent iron (nZVI) is recognized as a powerful tool for the remediation of groundwater contaminated by chlorinated ethenes (CEs). This long-term field study explored nZVI-driven degradation of CEs supported by electrokinetic (EK) treatment, which positively affects nZVI longevity and migration, and its impact on indigenous bacteria. In particular, the impact of combined nZVI-EK treatment on organohalide-respiring bacteria, ethenotrophs and methanotrophs (all capable of CE degradation) was assessed using molecular genetic markers detecting Dehalococcoides spp., Desulfitobacterium spp., the reductive dehalogenase genes vcrA and bvcA and ethenotroph and methanotroph functional genes. The remediation treatment resulted in a rapid decrease of the major pollutant cis-1,2-dichloroethene (cDCE) by 75% in the affected area, followed by an increase in CE degradation products methane, ethane and ethene. The newly established geochemical conditions in the treated aquifer not only promoted growth of organohalide-respiring bacteria but also allowed for the concurrent presence of vinyl chloride- and cDCE-oxidizing methanotrophs and (especially) ethenotrophs, which proliferated preferentially in the vicinity of an anode where low levels of oxygen were produced. The nZVI treatment resulted in a temporary negative impact on indigenous bacteria in the application well close to the cathode; but even there, the microbiome was restored within 15 days. The nZVI-EK treatment proved highly effective in reducing CE contamination and creating a suitable environment for subsequent biodegradation by changing groundwater conditions, promoting transport of nutrients and improving CE availability to soil and groundwater bacteria.