Bioremediation of diuron contaminated soils by a novel degrading microbial consortium.

Diuron is a biologically active pollutant present in soil, water and sediments. It is persistent in soil, water and groundwater and slightly toxic to mammals and birds as well as moderately toxic to aquatic invertebrates. Its principal product of biodegradation, 3,4-dichloroaniline, exhibits a higher toxicity than diuron and is also persistent in the environment. On this basis, the objective of the study was to determine the potential capacity of a proposed novel diuron-degrading microbial consortium (DMC) for achieving not only diuron degradation, but its mineralisation both in solution as well as in soils with different properties. The consortium was tested in a soil solution where diuron was the only carbon source, and more than 98.8% of the diuron initially added was mineralised after only a few days. The consortium was composed of three diuron-degrading strains, Arthrobacter sulfonivorans, Variovorax soli and Advenella sp. JRO, the latter had been isolated in our laboratory from a highly contaminated industrial site. This work shows for the first time the potential capacity of a member of the genus Advenella to remediate pesticide-contaminated soils. However, neither of the three strains separately achieved mineralisation (ring-14C) of diuron in a mineral medium (MSM) with a trace nutrient solution (NS); combined in pairs, they mineralised 40% of diuron in solution, but the most relevant result was obtained in the presence of the three-member consortium, where complete diuron mineralisation was achieved after only a few days. In the presence of the investigated soils in suspension, the capacity of the consortium to mineralise diuron was evaluated, achieving mineralisation of a wide range of herbicides from 22.9 to 69.0%.

Bioremediation of a trifluralin contaminated soil using bioaugmentation with novel isolated bacterial strains and cyclodextrin.

Trifluralin (TFL) is a highly persistent with a strong adsorption capacity on soil particles herbicide. This study was to isolate microbial consortia and bacterial strains from a soil with a historical application of pesticides to evaluate their potential to degrade TFL in soil. Different bioremediation techniques were considered for increasing the effectiveness of TFL degradation in soil. These techniques consisted of: i) biostimulation, using a nutrients solution (NS); ii) bioaugmentation, using a natural microbial consortium (NMC), seven individual bacterial strains isolated from NMC, and an artificial bacterial consortium formed by the seven TFL-degrading bacterial strains (ABC); iii) bioavailability enhancement, using a biodegradable compound, a randomly methylated cyclodextrin, RAMEB. Biostimulation using NS leads up to 34 % of soil TFL biodegraded after 100 d. When the contaminated soil was inoculated with NMC or ABC consortia, TFL loss increased up to 62 % and 74 %, respectively, with DT50 values (required time for the pollutant concentration to decline to half of its initial value) of 5.9 and 11 d. In the case of soil inoculation with the isolated individual bacterial strains, the extent of TFL biodegradation ranged widely from 2.3 % to 55 %. The most efficient bacterial strain was Arthrobacter aurescens CTFL7 which had not been previously described in the literature as a TFL-degrading bacterium. Bioaugmentation with CTFL7 bacterium was also tested in the presence of RAMEB, provoking a drastic increase in herbicide biodegradation up to 88 %, achieving a DT50 of only 19 d. Cyclodextrins had never been tested before for enhancement of TFL biodegradation. An ecotoxicity assay was performed to confirm that the proposed bioremediation techniques were also capable to reduce toxicity. A Microtox® test showed that after application A. aurescens CTF7 and A. aurescens CTF7 + RAMEB, the TFL-contaminated soil, which initially presented acute toxicity, became non-toxic at the end of the biodegradation experiments.

Dissipation of a mix of priority PAHs in soils by using availability enhancers. Effect of aging and pollutant interactions.

A remediation strategy using three non-toxic availability enhancers (two cyclodextrins and a rhamnolipid biosurfactant) was applied to various soils artificially contaminated with a mix of Polycyclic Aromatic Hydrocarbons (PAHs) considered priority pollutants at two levels of contamination: only with 7 low molecular weight PAHs (LMW PAHs, 5 with 3-ring and 2 with 4-ring - fluoranthene and pyrene) or with 14 PAHs (from 3 to 6 rings). Natural attenuation of PAHs in all soils showed degradation capacity for the LMW PAHs, with a final content of LMW PAHs <5% of their initial concentration. Conversely, the rest of PAHs (high molecular weight PAHs, HMW) remained in the soils (61% - 83.5%), indicating abiotic dissipation of HMW PAHs due to formation of non-extractable residues in soils. The influence of the presence of HMW PAHs on the degradation of the 7 LMW PAHs was also tested, showing a general decrease in the time to obtain 50% dissipation (DT50), statistically significant for acenaphthene, acenaphthylene and fluorene. Availability enhancers showed different effects on PAHs dissipation. 2-hydroxypropyl-ß-cyclodextrin (HP) decreased DT50 of some of the lighter PAHs, whereas the rhamnolipid (RL) caused a slight DT50 increase due to its initial toxicity on native soil microorganisms, but showing later high degradation rate for LMW PAHs. On the contrary, randomly methylated-ß-cyclodextrin (RAMEB) slowed down PAHs degradation due to its high adsorption onto soil surface, blocking the desorption of PAHs from the soils. The high number of experimental factors not studied simultaneously before (soil type, co-contamination, availability enhancers and incubation time) allowed to conduct a statistical analysis which supported the conclusions reached. Principal Component Analysis separated the studied PAHs in 3 groups, in relation with their molecular weight and Kow. The first principal component was related with LMW PAHs, and separate the inefficient RAMEB from the other availability enhancers.

Novel nonylphenol-degrading bacterial strains isolated from sewage sludge: Application in bioremediation of sludge.

Nonylphenol (NP) is an anthropogenic pollutant frequently found in sewage sludge due to the insufficient degrading effectiveness of conventional WWTPs and has attracted attention as an endocrine disruptor. The aim of this study was to isolate specific NP-degrading bacteria from sewage sludge to be used in the degradation of this contaminant through bioaugmentation processes in aqueous solution and sewage sludge. Up to eight different bacterial strains were isolated, six of them not previously described as NP degraders. Bacillus safensis CN12 presented the best NP degradation in solution, and glucose used as an external carbon source increased its effect, reaching DT50 degradation values (time to decline to half the initial concentration of the pollutant) of only 0.9 days and a complete degradation in <7 days. Four NP metabolites were identified throughout the biodegradation process, showing higher toxicity than the parent contaminant. In sewage sludge suspensions, the endogenous microbiota was capable of partially degrading NP, but a part remained adsorbed as bound residue. Bioaugmentation was used for the first time to remove NP from sewage sludge to obtain more environmentally friendly biosolids. However, B. safensis CN12 was not able to degrade NP due to its high adsorption on sludge, but the use of a cyclodextrin (HPBCD) as availability enhancer allowed us to extract NP and degrade it in solution. The addition of glucose as an external carbon source gave the best results since the metabolism of the sludge microbiota was activated, and HPBCD was able to remove NP from sewage sludge to the solution to be degraded by B. safensis CN12. These results indicate that B. safensis CN12 can be used to degrade NP in water and sewage sludge, but the method must be improved using consortia of B. safensis CN12 with other bacterial strains able to degrade the toxic metabolites produced.

Soil bioremediation by cyclodextrins. A review.

Remediation of soils contaminated by organic pollutants has become an urgent necessity worldwide. A wide variety of techniques have been developed but many of them are associated with drawbacks (complexity, high costs, environmental risks, etc.). Bioremediation, the use of living organisms to remediate polluted sites, is an alternative approach considered a cost-effective and more environmentally friendly technique, but the low bioavailability of the organic pollutants in soils is its main limitation. Cyclodextrins have been proposed as a ́greener alternative to organic solvents or synthetic surfactants for increasing organics bioavailability in soils. Cyclodextrins can form inclusion complexes with hydrophobic pollutants increasing their aqueous solubility and enhancing their bioremediation in soils. This review gives an overview on the use of cyclodextrins for this purpose, highlighting the advantages and disadvantages and perspectives of this technology for future research. The effect of those cyclodextrins more commonly used is analyzed, particularly hydroxypropyl-ß-cyclodextrin (HPBCD) and randomly methylated-ß-cyclodextrin (RAMEB), as well as some of the more common contaminants treated (almost 80% are industrial chemicals and the rest are pesticides) and the bioremediation strategies used (by microorganisms and/or phytoremediation). The review also provides a critical view on knowledge gaps and limitations of this technology which must be overcome to bring it for field-scale application.

QSAR/QSPR models based on quantum chemistry for risk assessment of pesticides according to current European legislation.

In Europe, agencies and official organizations involved in the pesticide control such as the EFSA, ECHA, JRC and ECETOC or even the OECD are pointing out that the software tools based on quantitative structure relationship models, i.e. QSAR and QSPR, have a huge potential to improve the pesticide risk assessment process. In this sense, these non-animal test methods can promote the competitiveness of agriculture in this region: the consumer safety is increased with them due to the possibility of perform an overall better risk assessment of the degradation products and metabolites from pesticides. However, the use of theses computational-based (in silico) tools must be much more systematised and harmonised, improving their validation and including case studies to test them. To open databases, incorporating critical data in an orderly manner for building the models, becomes also necessary. Moreover, quantum chemistry through the Density Functional Theory should be promoted as tool for calculation of quantum descriptors, especially for the study of similar compounds with the same carbon skeleton but differing substitution patterns, e.g. isomers.

Natural and assisted dissipation of polycyclic aromatic hydrocarbons in a long-term co-contaminated soil with creosote and potentially toxic elements.

An enhanced bioremediation strategy was applied to an industrial soil co-contaminated with Polycyclic Aromatic Hydrocarbons (PAHs) and Potentially Toxic Elements (PTEs). Hydroxypropyl-ß-cyclodextrin (HPBCD) and a natural mixture of two rhamnolipids (RL) were added to increase PAHs bioavailability, and combined with a microbial consortium (MC) to biodegrade soil PAHs. Bioavailability of only six PAHs (3-, 4-ring PAHs) increased when using HPBCD, with a maximum increase about 2.8-fold higher. The highest dose of HPBCD (5%) enhanced PAH degradation, with the best results for 4-ring PAHs with treatments of HPBCD + MC (up to 48% degradation for pyrene and 43% for fluoranthene), whereas dissipation for 5-ring PAHs was very low and for 6-ring was negligible. The use of RL increased the bioavailability of 13 of the 16 PAHs studied, reaching up to 60-fold higher values for phenanthrene or 18-fold higher for acenaphtene. RL addition did not show degradation improvement in any situation, and even inhibited the scarce degradation observed in the control treatment. The high increase in availability of both PAHs and mainly PTEs when using RL as amendment could make them toxic for microorganisms. In fact, Microtox Acute Toxicity test using Aliivibrio fischeri and the absence of colony forming units (CFUs) of indigenous bacteria demonstrated the extremely high levels of toxicity in RL treated soil.

Pulping cardoon (Cynara cardunculus) with peroxyformic acid (MILOX) in one single stage.

In this work, depithed cardoon stalk (Cynara cardunculus) has been used with the objective of obtaining bleachable pulps. The material, once properly prepared, was subjected to one-step peroxyformic acid delignification. In order to study the process, a face-centred second order factorial design was developed which allowed the determination of the influences of four variables: concentrations of formic acid and hydrogen peroxide in the cooking liquor and the time and temperature of the treatment. Empirical mathematical models have been obtained which predict the yield, kappa index, residual lignin content, and viscosity of the pulps. These models demonstrate that in general the delignification was extensive, producing pulps with kappa indexes less than 25 in the majority of cases, with good yields in the range of 45-60%. However, the pulps seem to have been degraded in the reaction media, as can be deduced from the low viscosity values found: 260-520 mL/g.

Combined use of microbial consortia isolated from different agricultural soils and cyclodextrin as a bioremediation technique for herbicide contaminated soils.

The phenylurea herbicide diuron is persistent in soil, water and groundwater and is considered to be a highly toxic molecule. The principal product of its biodegradation, 3,4-dichloroaniline, exhibits greater toxicity than diuron and is persistent in the environment. Five diuron degrading microbial consortia (C1C5), isolated from different agricultural soils, were investigated for diuron mineralization activity. The C2 consortium was able to mineralize 81.6% of the diuron in solution, while consortium C3 was only able to mineralize 22.9%. Isolated consortia were also tested in soil slurries and in all cases, except consortium C4, DT50 (the time required for the diuron concentration to decline to half of its initial value) was drastically reduced, from 700 days (non-inoculated control) to 546, 351, and 171 days for the consortia C5, C2, and C1, respectively. In order to test the effectiveness of the isolated consortium C1 in a more realistic scenario, soil diuron mineralization assays were performed under static conditions (40% of the soil water-holding capacity). A significant enhancement of diuron mineralization was observed after C1 inoculation, with 23.2% of the herbicide being mineralized in comparison to 13.1% for the control experiment. Hydroxypropyl-ß-cyclodextrin, a biodegradable organic enhancer of pollutant bioavailability, used in combination with C1 bioaugmentation in static conditions, resulted in a significant decrease in the DT50 (214 days; 881 days, control experiment). To the best of our knowledge, this is the first report of the use of soil-isolated microbial consortia in combination with cyclodextrins proposed as a bioremediation technique for pesticide contaminated soils.

Advanced technologies for the remediation of pesticide-contaminated soils.

The occurrence of pesticides in soil has become a highly significant environmental problem, which has been increased by the vast use of pesticides worldwide and the absence of remediation technologies that have been tested at full-scale. The aim of this review is to give an overview on technologies really studied and/or developed during the last years for remediation of soils contaminated by pesticides. Depending on the nature of the decontamination process, these techniques have been included into three categories: containment-immobilization, separation or destruction. The review includes some considerations about the status of emerging technologies as well as their advantages, limitations, and pesticides treated. In most cases, emerging technologies, such as those based on oxidation-reduction or bioremediation, may be incorporated into existing technologies to improve their performance or overcome limitations. Research and development actions are still needed for emerging technologies to bring them for full-scale implementation.