Effects of different organic substrate compositions on the decontamination of aged PAH-polluted soils through outdoor co-composting.

The effects of different organic substrate compositions on the efficiency of outdoor co-composting as a bioremediation technology for decontaminating soil polluted by polycyclic aromatic hydrocarbons (PAHs) were investigated. Four different substrate mixtures and two different aged PAH-contaminated soils were used in a semi-pilot-scale experiment that lasted nearly 700 days. The two soils (A and B) differed concerning both the initial concentrations of the Æ©16 US EPA PAHs (5926 vs. 369 mg kg-1, respectively) and the type of predominant PAH group by molecular weight. The experiments revealed that while the composition of the organic substrate had an impact on the rate of PAH degradation, it did not significantly influence the final extent of PAH degradation. Notably, the organic substrate consisting of green waste and wood chips (GW) was found to facilitate the most rapid rate of PAH degradation (first-order rate constant k = 0.033 ± 0.000 d-1 with soil A over the initial 42 days of the experiment and k = 0.036 ± 0.000 d-1 with soil B over the initial 56 days). Despite the differences in organic substrate compositions and types of soil being treated, PAH degradation levels exceeded at least 95% in all the treatments after more than 680 days of co-composting. Regardless of the composition, the removal of low- and medium- molecular-weight (2-4 rings) PAHs was nearly complete by the end of the experiment. Furthermore, high-molecular-weight PAHs (5 rings and more) were significantly degraded during co-composting, with reductions ranging from 54% to 79% in soil A and from 59% to 68% in soil B. All composts were dominated by Proteobacteria, Firmicutes, and Actinobacteria, with significant differences in abundance between soils. Genera with PAH degradation potentials were detected in all samples. The results of a battery of toxicity tests showed that there was almost no toxicity associated with the final composts.

New insights into vermiremediation of sewage sludge: The effect of earthworms on micropollutants and vice versa.

Vermicomposting represents an environmentally friendly method for the treatment of various types of biowastes, including sewage sludge (SS), as documented in numerous studies. However, there are few papers providing insights into the mechanisms and toxicity effects involved in SS vermicomposting to present a comprehensive overview of the process. In this work, the vermiremediation of SS containing various micropollutants, including pharmaceuticals, personal care products, endocrine disruptors, and per/polyfluoroalkyl substances, was studied. Two SSs originating from different wastewater treatment plants (WWTP1 and WWTP2) were mixed with a bulking agent, moistened straw, at ratios of 0, 25, 50, and 75% SS. Eisenia andrei earthworms were introduced into the mixtures, and after six weeks, the resulting materials were subjected to various types of chemical and toxicological analyses, including conventional assays (mortality, weight) as well as tissue- and cell-level assays, such as malondialdehyde production, cytotoxicity tests and gene expression assays. Through the vermiremediation process significant removal of diclofenac (90%), metoprolol (88%), telmisartan (62%), and triclosan (81%) was achieved. Although the concentrations of micropollutants were substantially different in the original SS samples, the micropollutants vermiaccumulated to a similar extent over the incubation period. The earthworms substantially eliminated the present bacterial populations, especially in the 75% SS treatments, in which the average declines were 90 and 79% for WWTP1 and WWTP2, respectively. To the best of our knowledge, this is the first study to investigate the vermiremediation of such a large group of micropollutants in real SS samples and provide a thorough evaluation of the effect of SS on earthworms at tissue and cellular level.

Solicola gregarius gen. nov., sp. nov., a soil actinobacterium isolated after enhanced cultivation with Micrococcus luteus culture supernatant.

An actinobacterial strain, designated A5X3R13T, was isolated from a compost soil suspension supplemented with extracellular material from a Micrococcus luteus-culture supernatant. The strain was cultured on tenfold-diluted reasoner's 2A agar. The cells were ovoid-to-rod shaped, non-motile, Gram-stain-positive, oxidase-negative, catalase-positive and had a width of 0.5 µm and a length of 0.8-1.2 µm. The results of both 16S rRNA-based phylogenetic and whole-genome analyses indicate that A5X3R13T forms a distinct lineage within the family Nocardioidaceae (order Propionibacteriales). On the basis of the 16S rRNA gene sequence, A5X3R13T was closely related to Aeromicrobium terrae CC-CFT486T (96.2 %), Nocardioides iriomotensis IR27-S3T (96.2 %), Nocardioides guangzhouensis 130T (95.6 %), Marmoricola caldifontis YIM 730233T (95.5 %), Aeromicrobium alkaliterrae KSL-107T (95.4 %), Aeromicrobium choanae 9H-4T (95.4 %), Aeromicrobium panaciterrae Gsoil 161T (95.3 %), and Nocardioides jensenii NBRC 14755T (95.2 %). The genome had a length of 4 915 757 bp, and its DNA G+C content was 68.5 mol %. The main fatty acids were 10-methyl C17 : 0, C16 : 0, C15 : 0, C18 : 0, C17 : 0 and iso-C16 : 0. The main polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol and two unidentified phospholipids. MK-9(H4) was the predominant respiratory quinone. The peptidoglycan type was A3γ (A41.1) and contained alanine, glycine, glutamic acid and ll-diaminopimelic acid in a molar ratio of 1.2 : 0.9 : 1.0 : 0.8. On the basis of the results of the phylogenetic and phenotypic analyses and comparisons with other members of the family Nocardioidaceae, strain A5X3R13T is proposed to represent a novel species within a novel genus, for which the name Solicola gregarius gen. nov., sp. nov. is proposed. The type strain is A5X3R13T (=DSM 112953T=NCCB 100840T).

Remedial trial of sequential anoxic/oxic chemico-biological treatment for decontamination of extreme hexachlorocyclohexane concentrations in polluted soil.

During production of γ-hexachlorocyclohexane (γ-HCH), thousands of tons of other isomers were synthesized as byproducts, and after dumping represent sources of contamination for the environment. Several microbes have the potential for aerobic and anaerobic degradation of HCHs, and zero-valent iron is an effective remediation agent for abiotic dechlorination of HCHs, whereas the combination of the processes has not yet been explored. In this study, a sequence of anoxic/oxic chemico-biological treatments for the degradation of HCHs in a real extremely contaminated soil (10-30 g/kg) was applied. Approximately 1500 kg of the soil was employed, and various combinations of reducing and oxygen-releasing chemicals were used for setting up the aerobic and anaerobic phases. The best results were obtained with mZVI/nZVI, grass cuttings, and oxygen-releasing compounds. In this case, 80 % removal of HCHs was achieved in 129 days, and 98 % degradation was achieved after 1106 days. The analysis of HCHs and their transformation products proved active degradation when slight accumulation of the transformation product during the anaerobic phase was followed by aerobic degradation. The results document that switching between aerobic and anaerobic phases, together with the addition of grass, also created suitable conditions for the biodegradation of HCHs and monochlorobenzene/benzene by microbes.

The driving factors of per- and polyfluorinated alkyl substance (PFAS) accumulation in selected fish species: The influence of position in river continuum, fish feed composition, and pollutant properties.

Per- and polyfluorinated alkyl substances (PFASs) represent a group of highly recalcitrant micropollutants, that continuously endanger the environment. The present work describes the geographical trends of fish contamination by individual PFASs (including new compounds, e.g., Gen-X) assessed by analyzing the muscle tissues of 5 separate freshwater fish species from 10 locations on the Czech section of the Elbe River and its largest tributary, the Vltava River. The data of this study also showed that the majority of the detected PFASs consisted of long-chain representatives (perfluorooctane sulfonate (PFOS), perfluorononanoic acid, perfluorodecanoic acid, and perfluoroundecanoic acid), whereas short-chain PFASs as well as other compounds such as Gen-X were detected in relatively small quantities. The maximum concentrations of the targeted 32 PFASs in fish were detected in the lower stretches of the Vltava and Elbe Rivers, reaching 289.9 ng/g dw, 140.5 ng/g dw, and 162.7 ng/g dw for chub, roach, and nase, respectively. Moreover, the relationships between the PFAS (PFOS) concentrations in fish muscle tissue and isotopic ratios (δ15N and δ13C) were studied to understand the effect of feed composition and position in the river continuum as a proxy for anthropogenic activity. Redundancy analysis and variation partitioning showed that the largest part of the data variability was explained by the interaction of position in the river continuum and δ15N (δ13C) of the fish. The PFAS concentrations increased downstream and were positively correlated with δ15N and negatively correlated with δ13C. A detailed study at one location also demonstrated the significant relationship between δ15N (estimated trophic position) and PFASs (PFOS) concentrations. From the tested physicochemical properties, the molecular mass and number of fluorine substituents seem to play crucial roles in PFAS bioaccumulation.

Discovering the potential of an nZVI-biochar composite as a material for the nanobioremediation of chlorinated solvents in groundwater: Degradation efficiency and effect on resident microorganisms.

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.

Polycyclic aromatic hydrocarbon accumulation in aged and unaged polyurethane microplastics in contaminated soil.

The interaction of microplastics (MPs) and common environmental organic pollutants has been a frequently discussed topic in recent years. Although the estimated contamination caused by MPs in terrestrial ecosystems is one order of magnitude higher than that in the oceans, experiments have been conducted solely in an aqueous matrix. Therefore, an experiment was carried out with two soils differing in their concentrations of polycyclic aromatic hydrocarbons (PAHs) and polyurethane foams used for scent fences along roads and crop fields. Two types of polyurethane foam (biodegradable and conventional in aged and unaged form) were exposed to soils containing PAHs that originated from historically contaminated localities. The exposure lasted 28 days, and a newly developed three-step procedure to separate MPs from soil was then applied. Biodegradable polyurethane MPs exhibited a strong tendency to accumulate PAHs after 7 days, and their concentrations significantly grew over time. In contrast, the sorption of PAHs on conventional polyurethane MPs was substantially lower (a maximum of 3.6 times higher concentration than that in the soil). Neither type of foam changed their sorption behaviors after the aging procedure. The results indicate that the flexibility of the polyurethane polymeric network could be the main driving factor for the sorption.

Screening for 32 per- and polyfluoroalkyl substances (PFAS) including GenX in sludges from 43 WWTPs located in the Czech Republic - Evaluation of potential accumulation in vegetables after application of biosolids.

Highly persistent, toxic and bioaccumulative per - and polyfluoroalkyl substances (PFAS) represents a serious problem for the environment and their concentrations and fate remain largely unknown. The present study consists of a PFAS screening in sludges originating from 43 wastewater treatment plants (WWTPs) in the Czech Republic. To analyze an extended group of PFAS consisting of 32 PFAS, including GenX and other new replacements of older and restricted PFAS in sludge, a new method was optimized and validated using pressurized solvent extraction, followed by the SPE clean-up step to eliminate the observed matrix effects and LC-MS/MS. The results revealed high PFAS contamination of sewage sludge, reaching values from 5.6 to 963.2 ng g-1. The results showed that in the majority of the samples (about 60%), PFOS was the most abundant among the targeted PFAS, reaching 932.9 ng g-1. Approximately 20% of the analyzed samples contained more short-chain PFAS, suggesting the replacement of long-chain PFAS (especially restricted PFOA and PFOS). GenX was detected in 9 samples, confirming the trend in the use of new PFAS. The results revealed that significantly higher contamination was detected in the samples from large WWTPs (population equivalent > 50,000; p-value <0.05). Concerning the application of sludge in agriculture, our prediction using the respective PFAS bioconcentration factors, the observed concentrations, and the legislatively permitted management of biosolids in Czech Republic agriculture revealed that PFAS can cause serious contamination of cereals and vegetables (oat, celery shoots and lettuce leaves), as well as general secondary contamination of the environment.

Biodegradation of PCBs in contaminated water using spent oyster mushroom substrate and a trickle-bed bioreactor.

Due to their persistence, polychlorinated biphenyls (PCBs) represent a group of important environmental pollutants, but conventional physicochemical decontamination techniques for their removal are usually expensive. The main aim of this work was to develop a cost-effective method for PCB bioremediation, focusing on contaminated water and utilizing the well-known degradation capability of Pleurotus ostreatus (the oyster mushroom). For this purpose, the conditions of several laboratory-scale reactors (working volume 1 L) were optimized. Spent oyster mushroom substrate obtained from a commercial farm was used as a fungal inoculum and growth substrate. The highest degradation efficiency (87%) was recorded with a continuous low-flow setup, which was subsequently scaled up (working volume 500 L) and used for the treatment of 4000 L of real contaminated groundwater containing 0.1-1 µg/L of PCBs. This trickle-bed pilot-scale bioreactor was able to remove 82, 80, 65, and 30-50% of di-, tri-, tetra- and pentachlorinated PCB congeners, respectively. No degradation was observed for hexa- or heptachlorinated congeners. Multiple mono- and dichlorobenzoic acids (CBAs) were identified as transformation products by mass spectrometry, confirming the role of biodegradation in PCB removal. A Vibrio fischeri bioluminescence inhibition test revealed slight ecotoxicity of the primary reactor effluent (sampling after 24 h), which was quickly suppressed once the effluent passed through the reactor for the second time. Moreover, no other effluent exhibited toxicity for the rest of the experiment (71 days in total). Microbial analyses (phospholipid fatty acid analysis and next-generation sequencing) showed that P. ostreatus was able to degrade PCBs in the presence of an abundance of other fungal species as well as aerobic and anaerobic bacteria. Overall, this study proved the suitability of the use of spent oyster mushroom substrate in a bioremediation practice, even for pollutants as recalcitrant as PCBs.

Oxidative stress in microbes after exposure to iron nanoparticles: analysis of aldehydes as oxidative damage products of lipids and proteins.

Due to their enhanced reactivity, metal and metal-oxide nanoscale zero-valent iron (nZVI) nanomaterials have been introduced into remediation practice. To ensure that environmental applications of nanomaterials are safe, their possible toxic effects should be described. However, there is still a lack of suitable toxicity tests that address the specific mode of action of nanoparticles, especially for nZVI. This contribution presents a novel approach for monitoring one of the most discussed adverse effects of nanoparticles, i.e., oxidative stress (OS). We optimized and developed an assay based on headspace-SPME-GC-MS analysis that enables the direct determination of volatile oxidative damage products (aldehydes) of lipids and proteins in microbial cultures after exposure to commercial types of nZVI. The method employs PDMS/DVB SPME fibers and pentafluorobenzyl derivatization, and the protocol was successfully tested using representatives of bacteria, fungi, and algae. Six aldehydes, namely, formaldehyde, acrolein, methional, benzaldehyde, glyoxal, and methylglyoxal, were detected in the cultures, and all of them exhibited dose-dependent sigmoidal responses. The presence of methional, which was detected in all cultures except those including an algal strain, documents that nZVI also caused oxidative damage to proteins in addition to lipids. The most sensitive toward nZVI exposure in terms of aldehyde production was the yeast strain Saccharomyces cerevisiae, which had an EC50 value of 0.08 g/L nZVI. To the best of our knowledge, this paper is the first to document the production of aldehydes resulting from lipids and proteins as a result of OS in microorganisms from different kingdoms after exposure to iron nanoparticles.

Implications of mycoremediated dry olive residue application and arbuscular mycorrhizal fungi inoculation on the microbial community composition and functionality in a metal-polluted soil.

Metal-polluted soils represent hostile environments affecting the composition and functions of soil microbial communities. This study evaluated the implication of combining the mycoremediated dry olive residue (MDOR) amendment application with the inoculation of the arbuscular mycorrhizal fungi (AMF) Funneliformis mosseae in restoring the quality, composition, and functionality of soil microbial communities. To achieve this aim, a mesocosms experiment was set up that included three variations: i) with and without application of Penicillium chrysogenum-10-transformed MDOR (MDOR_Pc), and Chondrosterum purpureum-transformed MDOR (MDOR_Cp) amendments; ii) with and without F. mosseae inoculation; and iii) 30-day and 60-day soil treatment time. As a result of this combined treatment, changes in the soil labile organic C and N fractions were observed throughout the experiment. Increases in the abundance of phospholipid fatty acids (PLFAs) for bacteria, actinobacteria, and Gram- and Gram+ bacteria were also recorded at the end of the experiment. The addition of MDOR amendments boosted fungal and AM fungi communities. AM fungi root and soil colonization was also enhanced as the result of improvement nutrient turnover and spatial conditions caused by adding MDOR in combination with an inoculation of F. mosseae. The composition and functionality of microbial communities seemed to be an important ecological attribute indicating an apparently fully functional restoration of this metal-polluted soil and therefore suggesting the suitability of the combined MDOR and AM fungus treatment as a reclamation practice.

Biodegradation of endocrine disruptors in urban wastewater using Pleurotus ostreatus bioreactor.

The white rot fungus Pleurotus ostreatus HK 35, which is also an edible industrial mushroom commonly cultivated in farms, was tested in the degradation of typical representatives of endocrine disrupters (EDCs; bisphenol A, estrone, 17ß-estradiol, estriol, 17α-ethinylestradiol, triclosan and 4-n-nonylphenol); its degradation efficiency under model laboratory conditions was greater than 90% within 12 days and better than that of another published strain P. ostreatus 3004. A spent mushroom substrate from a local farm was tested for its applicability in various batch and trickle-bed reactors in degrading EDCs in model fortified and real communal wastewater. The reactors were tested under various regimes including a pilot-scale trickle-bed reactor, which was finally tested at a wastewater treatment plant. The result revealed that the spent substrate is an efficient biodegradation agent, where the fungus was usually able to remove about 95% of EDCs together with suppression of the estrogenic activity of the sample. The results showed the fungus was able to operate in the presence of bacterial microflora in wastewater without any substantial negative effects on the degradation abilities. Finally, a pilot-scale trickle-bed reactor was installed in a wastewater treatment plant and successfully operated for 10days, where the bioreactor was able to remove more than 76% of EDCs present in the wastewater.

Assessment of biodegradation potential at a site contaminated by a mixture of BTEX, chlorinated pollutants and pharmaceuticals using passive sampling methods - Case study.

The present study describes a pilot remediation test of a co-mingled plume containing BTEX, chlorinated pollutants and pharmaceuticals. Remediation was attempted using a combination of various approaches, including a pump and treat system applying an advanced oxidation process and targeted direct push injections of calcium peroxide. The remediation process was monitored intensively and extensively throughout the pilot test using various conventional and passive sampling methods, including next-generation amplicon sequencing. The results showed that the injection of oxygen-saturated treated water with residual hydrogen peroxide and elevated temperature enhanced the in situ removal of monoaromatics and chlorinated pollutants. In particular, in combination with the injection of calcium peroxide, the conditions facilitated the in situ bacterial biodegradation of the pollutants. The mean groundwater concentration of benzene decreased from 1349µg·L-1 prior to the test to 3µg·L-1 within 3months after the calcium peroxide injections; additionally, monochlorobenzene decreased from 1545µg·L-1 to 36µg·L-1, and toluene decreased from 143µg·L-1 to 2µg·L-1. Furthermore, significant degradation of the contaminants bound to the soil matrix in less permeable zones was observed. Based on a developed 3D model, 90% of toluene and 88% of chlorobenzene bound to the soil were removed during the pilot test, and benzene was removed almost completely. On the other hand, the psychopharmaceuticals were effectively removed by the employed advanced oxidation process only from the treated water, and their concentration in groundwater remained stagnant due to inflow from the surroundings and their absence of in situ degradation. The employment of passive sampling techniques, including passive diffusion bags (PDB) for volatile organic pollutants and their respective transformation products, polar organic compound integrative samplers (POCIS) for the pharmaceuticals and in situ soil microcosms for microbial community analysis, was proven to be suitable for monitoring remediation in saturated zones.

Mesorhizobium bacterial strains isolated from the legume Lotus corniculatus are an alternative source for the production of polyhydroxyalkanoates (PHAs) to obtain bioplastics.

Polyhydroxyalkanoic acids (PHAs) are natural polyesters that can be used to produce bioplastics which are biodegradable. Numerous microorganisms accumulate PHAs as energy reserves. Combinations of different PHAs monomers lead to the production of bioplastics with very different properties. In the present work, we show the capability of strains belonging to various phylogenetic lineages within the genus Mesorhizobium, isolated from Lotus corniculatus nodules, to produce different PHA monomers. Among our strains, we found the production of 3-hydroxybutyrate, 3-hydroxyvalerate, 3-hydroxydodecanoate, and 3-hydroxyhexadecanoate. Most of the PHA-positive strains were phylogenetically related to the species M. jarvisii. However, our findings suggest that the ability to produce different monomers forming PHAs is strain-dependent.

Pharmaceuticals, benzene, toluene and chlorobenzene removal from contaminated groundwater by combined UV/H2O2 photo-oxidation and aeration.

This study was performed to test the feasibility of several decontamination methods for remediating heavily contaminated groundwater in a real contaminated locality in the Czech Republic, where a pharmaceuticals plant has been in operation for more than 80 years. The site is polluted mainly by recalcitrant psychopharmaceuticals and monoaromatic hydrocarbons, such as benzene, toluene and chlorobenzene. For this purpose, an advanced oxidation technique employing UV radiation with hydrogen peroxide dosing was employed, in combination with simple aeration pretreatment. The results showed that UV/H2O2 was an efficient and necessary step for degradation of the pharmaceuticals; however, the monoaromatics were already removed during the aeration step. Characterization of the removal mechanisms participating in the aeration revealed that volatilization, co-precipitation and biodegradation contributed to the process. These findings were supported by bacterial metabolite analyses, phospholipid fatty acid analysis, qPCR of representatives of the degradative genes and detailed characterization of the formed precipitate using Mössbauer spectroscopy and scanning electron microscopy. Further tests were carried out in a continuous arrangement directly connected to the wells already present in the locality. The results documented the feasibility of combination of the photo-reactor employing UV/H2O2 together with aeration pretreatment for 4 months, where the overall decontamination efficiency ranged from 72% to 99% of the pharmaceuticals. We recorded even better results for the monoaromatics decontamination except for one month, when we encountered some technical problems with the aeration pump. This demonstrated the necessity of using the aeration step.

Bioremediation of long-term PCB-contaminated soil by white-rot fungi.

The objective of this work was to test the PCB-degrading abilities of two white-rot fungi, namely Pleurotus ostreatus and Irpex lacteus, in real contaminated soils with different chemical properties and autochthonous microflora. In addition to the efficiency in PCB removal, attention was given to other important parameters, such as changes in the toxicity and formation of PCB transformation products. Moreover, structural shifts and dynamics of both bacterial and fungal communities were monitored using next-generation sequencing and phospholipid fatty acid analysis. The best results were obtained with P. ostreatus, which resulted in PCB removals of 18.5, 41.3 and 50.5% from the bulk, top (surface) and rhizosphere, respectively, of dumpsite soils after 12 weeks of treatment. Numerous transformation products were detected (hydoxylated and methoxylated PCBs, chlorobenzoates and chlorobenzyl alcohols), which indicates that both fungi were able to oxidize and decompose the aromatic moiety of PCBs in the soils. Microbial community analysis revealed that P. ostreatus efficiently colonized the soil samples and suppressed other fungal genera. However, the same fungus substantially stimulated bacterial taxa that encompass putative PCB degraders. The results of this study finally demonstrated the feasibility of using this fungus for possible scaled-up bioremediation applications.

Widely used pharmaceuticals present in the environment revealed as in vitro antagonists for human estrogen and androgen receptors.

A considerable amount of scientific evidence indicates that a number of pharmaceuticals that could be detected in the environment can contribute towards the development of problems associated with human reproductive systems, as well as those of wildlife. We investigated the estrogenic and androgenic effects of select pharmaceuticals with high production volume and environmental relevance. We examined the receptor-binding activities of these pharmaceuticals in the T47D human cell line using altered secretion of cytokine CXCL12. Functional yeast-luciferase reporter gene assays were also employed to confirm the mechanism of receptor binding by estrogen and androgen. Non-steroidal anti-inflammatory drugs, namely ibuprofen, diclofenac and antiarrhythmic agent amiodarone showed strong anti-estrogenic effects in the T47D cell line. In the yeast-luciferase assay, these anti-inflammatory drugs also demonstrated anti-estrogenic potency and inhibited the E2 response in a concentration-dependent manner. Amiodarone did not exhibit any response in the yeast-luciferase assay; therefore, the endocrine disruption presumably occurred at a different level without directly involving the receptor. All the anti-inflammatory drugs considered in this study, including ketoprofen, naproxen and clofibrate, exhibited a dose-dependent antagonism towards the androgen receptor in the yeast-luciferase assays. Several other drugs, including the stimulant caffeine, did not show any response in the tests that were employed. A risk assessment analysis using 'Hazard Quotient' suggested a potential risk, especially in the cases of ibuprofen, ketoprofen, diclofenac and clofibrate. The results reveal the intrinsic endocrine disrupting nature of several pharmaceuticals and thus could contribute towards explaining a number of adverse health effects on humans and wildlife.

Combined nano-biotechnology for in-situ remediation of mixed contamination of groundwater by hexavalent chromium and chlorinated solvents.

The present report describes a 13month pilot remediation study that consists of a combination of Cr(VI) (4.4 to 57mg/l) geofixation and dechlorination of chlorinated ethenes (400 to 6526µg/l), achieved by the sequential use of nanoscale zerovalent iron (nZVI) particles and in situ biotic reduction supported by whey injection. The remediation process was monitored using numerous techniques, including physical-chemical analyses and molecular biology approaches which enabled both the characterization of the mechanisms involved in pollutant transformation and the description of the overall background processes of the treatment. The results revealed that nZVI was efficient toward Cr(VI) by itself and completely removed it from the groundwater (LOQ 0.05mg/l) and the subsequent application of whey resulted in a high removal of chlorinated ethenes (97 to 99%). The persistence of the reducing conditions, even after the depletion of the organic substrates, indicated a complementarity between nZVI and the whey phases in the combined technology as the subsequent application of whey phase partially assisted the microbial regeneration of the spent nZVI by promoting its reduction into Fe(II), which further supported remediation conditions at the site. Illumina sequencing and the detection of functional vcrA and bvcA genes documented a development in the reducing microbes (iron-reducing, sulfate-reducing and chlororespiring bacteria) that benefited under the conditions of the site and that was probably responsible for the high dechlorination and/or Cr(VI) reduction. The results of this study demonstrate the feasibility and high efficiency of the combined nano-biotechnological approach of nZVI and whey application in-situ for the removal of Cr(VI) and chlorinated ethenes from the groundwater of the contaminated site.

Polycyclic aromatic hydrocarbons degradation and microbial community shifts during co-composting of creosote-treated wood.

The feasibility of decontaminating creosote-treated wood (CTW) by co-composting with agricultural wastes was investigated using two bulking agents, grass cuttings (GC) and broiler litter (BL), each employed at a 1:1 ratio with the matrix. The initial concentration of total polycyclic aromatic hydrocarbons (PAHs) in CTW (26,500 mg kg(-1)) was reduced to 3 and 19% after 240 d in GC and BL compost, respectively. PAH degradation exceeded the predicted bioaccesible threshold, estimated through sequential supercritical CO2 extraction, together with significant detoxification, assessed by contact tests using Vibrio fisheri and Hordeum vulgare. GC composting was characterized by high microbial biomass growth in the early phases, as suggested by phospholipid fatty acid analyses. Based on the 454-pyrosequencing results, fungi (mostly Saccharomycetales) constituted an important portion of the microbial community, and bacteria were characterized by rapid shifts (from Firmicutes (Bacilli) and Actinobacteria to Proteobacteria). However, during BL composting, larger amounts of prokaryotic and eukaryotic PLFA markers were observed during the cooling and maturation phases, which were dominated by Proteobacteria and fungi belonging to the Ascomycota and those putatively related to the Glomeromycota. This work reports the first in-depth analysis of the chemical and microbiological processes that occur during the co-composting of a PAH-contaminated matrix.

Combined abiotic and biotic in-situ reduction of hexavalent chromium in groundwater using nZVI and whey: A remedial pilot test.

The paper describes a pilot remediation test combining two Cr(VI) geofixation methods - chemical reduction by nanoscale zero-valent iron (nZVI) and subsequent biotic reduction supported by whey. Combination of the methods exploited the advantages of both - a rapid decrease in Cr(VI) concentrations by nZVI, which prevented further spreading of the contamination and facilitated subsequent use of the cheaper biological method. Successive application of whey as an organic substrate to promote biotic reduction of Cr(VI) after application of nZVI resulted in a further and long-term decrease in the Cr(VI) contents in the groundwater. The effect of biotic reduction was observed even in a monitoring well located at a distance of 22 m from the substrate injection wells after 10 months. The results indicated a reciprocal effect of both the phases - nZVI oxidized to Fe(III) during the abiotic phase was microbially reduced back to Fe(II) and acted as a reducing agent for Cr(VI) even when the microbial density was already low due to the consumed substrate. Community analysis with pyrosequencing of the 16S rRNA genes further confirmed partial recycling of nZVI in the form of Fe(II), where the results showed that the Cr(VI) reducing process was mediated mainly by iron-reducing and sulfate-reducing bacteria.