Irma S. Morelli

Effects of the Inoculant Strain Sphingomonas paucimobilis 20006FA on Soil Bacterial Community and Biodegradation in Phenanthrene-contaminated Soil

The effects of the inoculant strain Sphingomonas paucimobilis 20006FA (isolated from a phenanthrene-contaminated soil) on the dynamics and structure of microbial communities and phenanthrene elimination rate were studied in soil microcosms artificially contaminated with phenanthrene. The inoculant managed to be established from the first inoculation as it was evidenced by denaturing gradient gel electrophoresis analysis, increasing the number of cultivable heterotrophic and PAH-degrading cells and enhancing phenanthrene degradation. These effects were observed only during the inoculation period. Nevertheless, the soil biological activity (dehydrogenase activity and CO2 production) showed a late increase. Whereas gradual and successive changes in bacterial community structures were caused by phenanthrene contamination, the inoculation provoked immediate, significant, and stable changes on soil bacterial community. In spite of the long-term establishment of the inoculated strain, at the end of the experiment, the bioaugmentation did not produce significant changes in the residual soil phenanthrene concentration and did not improve the residual effects on the microbial soil community.

Remediation of phenanthrene-contaminated soil by simultaneous persulfate chemical oxidation and biodegradation processes

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous compounds with carcinogenic and/or mutagenic potential. To address the limitations of individual remediation techniques and to achieve better PAH removal efficiencies, the combination of chemical and biological treatments can be used. The degradation of phenanthrene (chosen as a model of PAH) by persulfate in freshly contaminated soil microcosms was studied to assess its impact on the biodegradation process and on soil properties. Soil microcosms contaminated with 140xa0mg/kgDRY SOIL of phenanthrene were treated with different persulfate (PS) concentrations 0.86–41.7xa0g/kgDRY SOIL and incubated for 28xa0days. Analyses of phenanthrene and persulfate concentrations and soil pH were performed. Cultivable heterotrophic bacterial count was carried out after 28xa0days of treatment. Genetic diversity analysis of the soil microcosm bacterial community was performed by PCR amplification of bacterial 16S rDNA fragments followed by denaturing gradient gel electrophoresis (DGGE). The addition of PS in low concentrations could be an interesting biostimulatory strategy that managed to shorten the lag phase of the phenanthrene biological elimination, without negative effects on the physicochemical and biological soil properties, improving the remediation treatment.

Monitoring the impact of bioaugmentation with a PAH-degrading strain on different soil microbiomes using pyrosequencing

The effect of bioaugmentation with Sphingobium sp. AM strain on different soils microbiomes, pristine soil (PS), chronically contaminated soil (IPK) and recently contaminated soil (Phe) and their implications in bioremediation efficiency was studied by focusing on the ecology that drives bacterial communities in response to inoculation. AM strain draft genome codifies genes for metabolism of aromatic and aliphatic hydrocarbons. In Phe, the inoculation improved the elimination of phenanthrene during the whole treatment, whereas in IPK no improvement of degradation of any PAH was observed. Through the pyrosequencing analysis, we observed that inoculation managed to increase the richness and diversity in both contaminated microbiomes, therefore, independently of PAH degradation improvement, we observed clues of inoculant establishment, suggesting it may use other resources to survive. On the other hand, the inoculation did not influence the bacterial community of PS. On both contaminated microbiomes, incubation conditions produced a sharp increase on Actinomycetales and Sphingomonadales orders, while inoculation caused a relative decline of Actinomycetales. Inoculation of most diverse microbiomes, PS and Phe, produced a coupled increase of Sphingomonadales, Burkholderiales and Rhizobiales orders, although it may exist a synergy between those genera; our results suggest that this would not be directly related to PAH degradation.

Autochthonous bioaugmentation to enhance phenanthrene degradation in soil microcosms under arid conditions

AbstractnThe aim of this work was to investigate the effect of autochthonous bioaugmentation (ABA) in phenanthrene-contaminated Patagonian soil microcosms, maintained under arid conditions, on phenanthrene elimination and soil microbial community. The polycyclic aromatic hydrocarbon (PAH)-degrading strain Sphingobium sp. 22B previously isolated from the Patagonian soil and selected by its resistance to drying conditions was used as inoculant. The phenanthrene concentration, dehydrogenase activity and denaturing gradient electrophoresis of 16S rRNA gene were monitored during 230xa0days. The results showed that when the microcosms were maintained at 20xa0% of soil water-holding capacity (WHC), the phenanthrene biodegradation was drastically inhibited and changes in the genetic diversity of soil microbial community were not detected, and neither the ABA nor the biostimulation managed to overcome the inhibitory effects. When the moisture was slightly increased, reached 25xa0% WHC, the ABA showed a significant initial stimulatory effect on phenanthrene biodegradation, demonstrating the potential of ABA in PAH bioremediation process in semiarid Patagonia.

Assessment of the Responsiveness to Different Stresses of the Microbial Community from Long-Term Hydrocarbon-Contaminated Soils

Soils exposed to long-term contamination with hydrocarbons may present extreme challenges to maintain the biological resilience to the stress. To elucidate the relationships between the initial event of contamination and the responsiveness to the stress, we investigated the extent of the microbial resilience of biological functions from two contaminated soils sampled from a petrochemical area (S1, underwent diffuse hydrocarbon contamination, and S2, from a land farming unit where an alkaline petrochemical sludge was treated) after the Cd, saline, and acid stresses. Both contaminated soils were characterized by low organic matter content compared with a pristine soil. Although similar Shannon diversity index and heterotrophic bacterial count were observed, different bacterial community structures (PCR-DGGE) and less enzymatic activities characterized the contaminated soils. Particularly, functional diversity determined by Biolog EcoPlates™ was not detected in S2 soil. Only the S1 soil showed resilience of the enzymatic activities and functional diversity, suggesting the presence of a well-adapted microbial community able to face with the stresses. The S2 was the most disturbed and less responsive soil. However, an increase in the functional diversity was evidenced after acidification, and it is possible to correlate this responsiveness with the sludge properties treated in the land farming unit. In addition, if the selected stress can reverse the soil condition provoked for the first disturbance, responsiveness could be expected.

Efficiency of surfactant-enhanced bioremediation of aged polycyclic aromatic hydrocarbon-contaminated soil: Link with bioavailability and the dynamics of the bacterial community

Shifts in the bacterial-community dynamics, bioavailability, and biodegradation of polycyclic aromatic hydrocarbons (PAHs) of chronically contaminated soil were analyzed in Triton X-100-treated microcosms at the critical micelle concentration (T-CMC) and at two sub-CMC doses. Only the sub-CMC-dose microcosms reached sorbed-PAH concentrations significantly lower than the control: 166±32 and 135±4mgkg-1 dry soil versus 266±51mgkg-1; consequently an increase in high- and low-molecular-weight PAHs biodegradation was observed. After 63days of incubation pyrosequencing data evidenced differences in diversity and composition between the surfactant-modified microcosms and the control, with those with sub-CMC doses containing a predominance of the orders Sphingomonadales, Acidobacteriales, and Gemmatimonadales (groups of known PAHs-degrading capability). The T-CMC microcosm exhibited a lower richness and diversity index with a marked predominance of the order Xanthomonadales, mainly represented by the Stenotrophomonas genus, a PAHs- and Triton X-100-degrading bacterium. In the T-CMC microcosm, whereas the initial surface tension was 35mNm-1, after 63days of incubation an increase up to 40mNm-1 was registered. The previous observation and the gas-chromatography data indicated that the surfactant may have been degraded at the CMC by a highly selective bacterial community with a consequent negative impact on PAHs biodegradation. This work obtained strong evidence for the involvement of physicochemical and biologic influences determining the different behaviors of the studied microcosms. The results reported here contribute significantly to an optimization of, surfactant-enhanced bioremediation strategies for chronically contaminated soil since the application of doses below the CMC would reduce the overall costs.

Insights into the mechanisms of desiccation resistance of the Patagonian PAH-degrading strain Sphingobium sp. 22B

To analyse the physiological response of Sphingobium sp. 22B to water stress.

Strategies for oxidation of PAHs in aged contaminated soil by batch reactors

Polycyclic aromatic hydrocarbons (PAH) are neutral, nonpolar and hydrophobic molecules that tend to sorb onto soil organic matter. Chemical oxidation is a good choice to avoid the limitations of bioremediation. To evaluate the efficiency of different types of oxidation (permanganate, hydrogen peroxide, and persulfate) and activation (heat, alkaline, and iron), batch reactors were prepared. The soil was contaminated with phenanthrene and pyrene (1200 ± 200 and 2800 ± 100mg per kg of dry soil, respectively) and aged for fifteen months. Treatments were prepared with 10g of contaminated dry soil and 20ml of water and incubated at room temperature for 7 days. Analyses of phenanthrene and pyrene concentrations, soil pH and electric conductivity were performed. Counts of heterotrophic cultivable bacteria on R2A medium and PAH-degraders were carried out after 7 days of treatment. The persulfate treatment at room temperature, without the addition of activators, achieved better results than treatments with the same doses of permanganate or hydrogen peroxide. All the strategies to improve persulfate treatments yielded higher degradation of pyrene than the biological control, as expected from the structural description of this compound by Clars model. The thermal activation of persulfate (65°C for 6h) led to the degradation of more than 90% of both PAHs after 7 days of treatment.