Soil amendment and rhizobacterial inoculation improved Cu phytostabilization, plant growth and microbial activity in a bench-scale experiment.

Mine driven trace elements' pollution entails environmental risks and causes soil infertility. In the last decades, in situ techniques such as phytostabilization have become increasingly important as ways to tackle these negative impacts. The aim of this study was to test the individual and combined effects of different aided phytostabilization techniques using substrate from barren tailings of a Cu mine, characterized by extreme infertility (high acidity and deficiency of organic matter and nutrients). The experiment analyzed the growth of Populus nigra L. planted alone (P) or in co-cropping with Trifolium repens L. (PT), in pots containing mine soil amended with compost (1, 10, compost, soil, w/w) non inoculated (NI) or inoculated with plant growth promoting rhizobacteria (PGP), mycorrhizae (MYC) or a combination of bacterial and fungal inocula (PGPMYC). Non-amended, non-planted and non-inoculated reference ports were also prepared. Plants were harvested after 110 days of plant development and several biometric and phytopathological parameters (stem height, aerial biomass, root biomass, wilting, chlorosis, pest and death) and macro and micronutrient composition were determined. The growth substrate was analyzed for several physicochemical (pH, CECe, and exchangeable cations, total C and N, P Olsen and availability of trace elements) and microbiological (community level physiological profiles: activity, richness and diversity) parameters. The use of the amendment, P. nigra plantation, and inoculation with rhizobacteria were the best techniques to reduce toxicity and improve soil fertility, as well as to increase the plant survival and growth. Soil bacterial functional diversity was markedly influenced by the presence of plants and the inoculation with bacteria, which suggests that the presence of plant regulated the configuration of a microbial community in which the inoculated bacteria thrive comparatively better. The results of this study support the use of organic amendments, tolerant plants, and plant growth promoting rhizobacteria to reduce environmental risk and improve fertility of soils impacted by mining.

Effect of plant root exudates on the desorption of hexachlorocyclohexane isomers from contaminated soils.

Plants and their associated microbiota can have a significant impact on the behaviour of soil contaminants. Particularly, root exudation is one of the most important plant-associated processes in this respect, as it may have a substantial effect on the bioavailability of soil contaminants, specially of hydrophobic contaminants strongly sorbed by soil. The aim of the present study was to evaluate the effect of root exudates (natural and artificial) on the desorption of α-, ß-, δ- and γ-isomers of hexachlorocyclohexane (HCH) from contaminated soil, using batch experiments. Natural root exudates were obtained from Holcus lanatus plants growing in the same (contaminated) area. Fifteen compounds (mainly organic acids and phenolic compounds) usually found in root exudates were also tested, individually or as mixtures (1 and 10 mM). Both natural and artificial exudates favoured the mobilization of sorbed HCH in soil. The effect was highly significant for α-, ß- and γ-HCH isomers, for which the desorption rates increased by 23.0, 26.8 and 15.5% in the presence of natural root exudates and by 40.1, 25.9 and 25.6% in the presence of the artificial mixture (at 10 mM). The δ-HCH desorption rates increased by less than 10%. The effect of individual exudate components was very variable, but increased with the carbon content, reflecting the significance of hydrophobic interactions between the exudates and HCH molecules in the desorption of these last from soil. These findings indicate that plants may significantly influence the bioavailability of persistent contaminants, with major implications for improving phyto- and bioremediation procedures.

Modelling the ex situ bioremediation of diesel-contaminated soil in a slurry bioreactor using a hydrocarbon-degrading inoculant.

Bioremediation is a soil clean-up technique which exploits the metabolic capacity of microorganisms to degrade soil contaminants. A model was developed to simulate the ex situ bioremediation of a diesel-contaminated soil in a bio-slurry reactor inoculated with a diesel-degrading bacterial strain. Mass transfer processes involving desorption of diesel from soil to water and volatilization of diesel from water, and biodegradation by the bacterial inoculant were included in the model by using Weibull sigmoid kinetics and logistic/Monod kinetics respectively. Model parameters were estimated in batch-based abiotic and biodegradation experiments. Sensitivity analysis revealed the importance of maintaining a high bacterial density in the system for maximum bioremediation efficiency. The model was validated using a pilot bioreactor monitored for 528 h, which removed almost 90% of the diesel present in the system. The results revealed the capacity of the model to predict the bioremediation efficiency under different scenarios by adapting the input parameters to each system.

Characterization and degradation potential of diesel-degrading bacterial strains for application in bioremediation.

Bioremediation of polluted soils is a promising technique with low environmental impact, which uses soil organisms to degrade soil contaminants. In this study, 19 bacterial strains isolated from a diesel-contaminated soil were screened for their diesel-degrading potential, biosurfactant (BS) production, and biofilm formation abilities, all desirable characteristics when selecting strains for re-inoculation into hydrocarbon-contaminated soils. Diesel-degradation rates were determined in vitro in minimal medium with diesel as the sole carbon source. The capacity to degrade diesel range organics (DROs) of strains SPG23 (Arthobacter sp.) and PF1 (Acinetobacter oleivorans) reached 17-26% of total DROs after 10 days, and 90% for strain GK2 (Acinetobacter calcoaceticus). The amount and rate of alkane degradation decreased significantly with increasing carbon number for strains SPG23 and PF1. Strain GK2, which produced BSs and biofilms, exhibited a greater extent, and faster rate of alkane degradation compared to SPG23 and PF1. Based on the outcomes of degradation experiments, in addition to BS production, biofilm formation capacities, and previous genome characterizations, strain GK2 is a promising candidate for microbial-assisted phytoremediation of diesel-contaminated soils. These results are of particular interest to select suitable strains for bioremediation, not only presenting high diesel-degradation rates, but also other characteristics which could improve rhizosphere colonization.

Use of plant growth promoting bacterial strains to improve Cytisus striatus and Lupinus luteus development for potential application in phytoremediation.

Plant growth promoting (PGP) bacterial strains possess different mechanisms to improve plant development under common environmental stresses, and are therefore often used as inoculants in soil phytoremediation processes. The aims of the present work were to study the effects of a collection of plant growth promoting bacterial strains on plant development, antioxidant enzyme activities and nutritional status of Cytisus striatus and/or Lupinus luteus plants a) growing in perlite under non-stress conditions and b) growing in diesel-contaminated soil. For this, two greenhouse experiments were designed. Firstly, C. striatus and L. luteus plants were grown from seeds in perlite, and periodically inoculated with 6 PGP strains, either individually or in pairs. Secondly, L. luteus seedlings were grown in soil samples of the A and B horizons of a Cambisol contaminated with 1.25% (w/w) of diesel and inoculated with best PGP inoculant selected from the first experiment. The results indicated that the PGP strains tested in perlite significantly improved plant growth. Combination treatments provoked better growth of L. luteus than the respective individual strains, while individual inoculation treatments were more effective for C. striatus. L. luteus growth in diesel-contaminated soil was significantly improved in the presence of PGP strains, presenting a 2-fold or higher increase in plant biomass. Inoculants did not provoke significant changes in plant nutritional status, with the exception of a subset of siderophore-producing and P-solubilising bacterial strains that resulted in significantly modification of Fe or P concentrations in leaf tissues. Inoculants did not cause significant changes in enzyme activities in perlite experiments, however they significantly reduced oxidative stress in contaminated soils suggesting an improvement in plant tolerance to diesel. Some strains were applied to non-host plants, indicating a non-specific performance of their plant growth promotion. The use of PGP strains in phytoremediation may help plants to overcome contaminant and other soil stresses, increasing phytoremediation efficiency.

Enhanced Degradation of Diesel in the Rhizosphere of after Inoculation with Diesel-Degrading and Plant Growth-Promoting Bacterial Strains.

The association of plants and rhizospheric bacteria provides a successful strategy to clean up contaminated soils. The purpose of this work was to enhance diesel degradation in rhizosphere by inoculation with selected bacterial strains: a diesel degrader (D), plant growth-promoting (PGP) strains, or a combination (D+PGP). Plants were set up in pots with the A or B horizon of an umbric Cambisol (A and B) spiked with diesel (1.25%, w/w). After 1 mo, the dissipation of diesel range organics (DRO) with respect to = 0 (i.e., 1 wk after preparing the pots with the seedlings) concentration was significantly higher in inoculated than in noninoculated (NI) pots: The highest DRO losses were found in A D+PGP pots (close to 15-20% higher than NI) and in B D pots (close to 10% higher). The water-extractable DRO fraction was significantly higher at = 30 d (15-25%) compared with = 0 (<5%), probably due to the effects of plant root exudates and biosurfactants produced by the degrader strain. The results of this experiment reflect the importance of the partnerships between plants and bacterial inoculants and demonstrate the relevance of the effect of bacterial biosurfactants and plant root exudates on contaminant bioavailability, a key factor for enhancing diesel rhizodegradation. The association of lupine with D and PGP strains resulted in a promising combination for application in the rhizoremediation of soils with moderate diesel contamination.

Characterization and fingerprinting of soil and groundwater contamination sources around a fuel distribution station in Galicia (NW Spain).

Soil and groundwater contamination around a fuel distribution station in Tomiño (NW Spain) was evaluated. For this purpose, top and subsoil (up to 6.4 m) and groundwater were sampled around the station, approximately in a 60-m radius. Samples were analysed by HS-SPME-GC-MS to identify and quantify volatile fuel organic compounds (VFOC) (MTBE, ETBE and BTEX) and diesel range organics (DRO). Analysis and fingerprinting data suggested that the contamination of soil and groundwater was provoked by a fuel leak from underground storage tanks. This was reflected by hydrocarbon indices and principal component analysis, which discriminated a direct source of contamination of the subsoil samples around the station. The contaminants probably migrated from tank nearby soils to surrounding soils and leached to groundwater, following a SW direction. Irrigation with contaminated groundwater provoked a severe contamination of topsoils, which were enriched with the lightest components of gasoline and diesel. Fingerprinting also revealed the continuity of the leak, reflected by the presence of volatiles in some samples, which principally appeared in fresh leaks. MTBE was detected in a very high concentration in groundwater samples (up to 690 µg L(-1)), but it was not detected in fresh gasoline. This also evidenced an old source of contamination, probably starting in the mid-1990s, when the use of MTBE in gasoline was regulated.

Leachability of volatile fuel compounds from contaminated soils and the effect of plant exudates: A comparison of column and batch leaching tests.

Volatile fuel compounds such as fuel oxygenates (FO) (MTBE and ETBE) and BTEX (benzene, toluene, ethylbenzene and xylene) are some of the most soluble components of fuel. Characterizing the leaching potential of these compounds is essential for predicting their mobility through the soil profile and assessing the risk of groundwater contamination. Plant root exudates can play an important role in the modification of contaminant mobility in soil-plant systems, and such effects should also be considered in leaching studies. Artificially spiked samples of A and B horizons from an alumi-umbric Cambisol were leached in packed-columns and batch experiments using Milli-Q water and plant root exudates as leaching agents. The leaching potential and rate were strongly influenced by soil-contaminant interactions and by the presence of root exudates. Organic matter in A horizon preferably sorbed the most non-polar contaminants, lowering their leaching potential, and this effect was enhanced by the presence of root exudates. On the other hand, the inorganic components of the B horizon, showed a greater affinity for polar molecules, and the presence of root exudates enhanced the desorption of the contaminants. Column experiments resulted in a more realistic protocol than batch tests for predicting the leaching potential of volatile organic compounds in dissimilar soils.