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Polar biotransformation products have been identified as causative agents for the eventual increase in genotoxicity observed after the bioremediation of PAH-contaminated soils. Their further biodegradation has been described under certain biostimulation conditions; however, the underlying microorganisms and mechanisms remain to be elucidated. 9,10-Anthraquinone (ANTQ), a transformation product from anthracene (ANT), is the most commonly detected oxygenated PAH (oxy-PAH) in contaminated soils. Sand-in-liquid microcosms inoculated with creosote-contaminated soil revealed the existence of a specialized ANTQ degrading community, and Sphingobium sp. AntQ-1 was isolated for its ability to grow on this oxy-PAH. Combining the metabolomic, genomic, and transcriptomic analyses of strain AntQ-1, we comprehensively reconstructed the ANTQ biodegradation pathway. Novel mechanisms for polyaromatic compound degradation were revealed, involving the cleavage of the central ring catalyzed by Baeyer-Villiger monooxygenases (BVMO). Abundance of strain AntQ-1 16S rRNA and its BVMO genes in the sand-in-liquid microcosms correlated with maximum ANTQ biodegradation rates, supporting the environmental relevance of this mechanism. Our results demonstrate the existence of highly specialized microbial communities in contaminated soils responsible for processing oxy-PAHs accumulated by primary degraders. Also, they underscore the key role that BVMO may play as a detoxification mechanism to mitigate the risk posed by oxy-PAH formation during bioremediation of PAH-contaminated soils.

We studied how sunflower plants affect rhamnolipid biosurfactant mobilization of slowly desorbing fractions of polycyclic aromatic hydrocarbons (PAHs) in soil from a creosote-contaminated site. Desorption kinetics of 13 individual PAHs revealed that the soil contained initially up to 50% slowly desorbing fractions. A rhamnolipid biosurfactant was applied to the soil at the completion of the sunflower cycle (75 days in greenhouse conditions). After this period, the PAHs that remained in the soil were mainly present in a slowly desorbing form as a result of the efficient biodegradation of fast-desorbing PAHs by native microbial populations. The rhamnolipid enhanced the bioavailable fraction of the remaining PAHs by up to 30%, as evidenced by a standardized desorption extraction with Tenax, but the enhancement occurred with only planted soils. The enhanced bioavailability did not decrease residual PAH concentrations under greenhouse conditions, possibly due to ecophysiological limitations in the biodegradation process that were independent of the bioavailability. However, biodegradation was enhanced during slurry treatment of greenhouse planted soils that received the biosurfactant. The addition of rhamnolipids caused a dramatic shift in the soil bacterial community structure, which was magnified in the presence of sunflower plants. The stimulated groups were identified as fast-growing and catabolically versatile bacteria. This new rhizosphere microbial biomass possibly interacted with the biosurfactant to facilitate intra-aggregate diffusion of PAHs, thus enhancing the kinetics of slow desorption. Our results show that the usually limited biosurfactant efficiency with contaminated field soils can be significantly enhanced by integrating the sunflower ontogenetic cycle into the bioremediation design.

The application of a rhamnolipid biosurfactant for enhanced solubilization and biodegradation of slowly desorbing polycyclic aromatic hydrocarbons (PAHs) in contaminated soils was determined in this study. The soil samples exhibited different levels of pollution and different bioremediation stages: the first soil originated from a creosote-polluted site, contained 4370 mg kg −1 of PAHs and had not been bioremediated; the second soil was the same as the first but had received bioremediation treatment with nutrient amendment in biopiles for a period of 5 months and contained 580 mg kg −1 of PAHs after this treatment; the third soil was treated by bioremediation for several years to reduce the concentration of PAHs to 275 mg kg −1 . The kinetics of PAH desorption were determined to assess the magnitude of the slowly desorbing fractions present in the polluted soil and to optimize the biosurfactant effectiveness in terms of biodegradation. The soils that had been treated by bioremediation were enriched in slowly desorbing PAHs. The rhamnolipid at a concentration above its critical micelle concentration enhanced biodegradation in the soils that had been bioremediated previously. The measurement of residual concentrations of native PAHs showed the promoting effect of the biosurfactant on the biodegradation of the slowly desorbing fractions. Interestingly, benzo(a)pyrene was biodegraded in the soil that had been bioremediated for a long time. Rhamnolipid can constitute a valid alternative to chemical surfactants in promoting the biodegradation of slow-desorption PAHs, which is one of the most important problems in bioremediation, but the efficiency depends strongly on the bioremediation stage in which the biosurfactant is applied

This work examines the role of mycelia in enhancing the degradation by attached bacteria of organic pollutants that have poor bioavailability. Two oomycetes, Pythium oligandrum and Pythium aphanidermatum, were selected as producers of mycelial networks, while Mycobacterium gilvum VM552 served as a model polycyclic aromatic hydrocarbon (PAH) degrading bacterium. The experiments consisted of bacterial cultures exposed to a nondisturbed nonaqueous phase liquid (NAPL) layer containing a heavy fuel spiked with 14C-labeled phenanthrene that were incubated in the presence or absence of the mycelia of the oomycetes in both shaking and static conditions. At the end of the incubation, the changes in the total alkane and PAH contents in the NAPL residue were quantified. The results revealed that with shaking and the absence of mycelia, the strain VM552 grew by utilizing the bulk of alkanes and PAHs in the fuel; however, biofilm formation was incipient and phenanthrene was mineralized following zero-order kinetics, due to bioavailability limitations. The addition of mycelia favored biofilm formation and dramatically enhanced the mineralization of phenanthrene, up to 30 times greater than the rate without mycelia, possibly by providing a physical support to bacterial colonization and by supplying nutrients at the NAPL/water interface. The results in the static condition were very different because the bacterial strain alone degraded phenanthrene with sigmoidal kinetics but could not degrade alkanes or the bulk of PAHs. We suggest that bacteria/oomycete interactions should be considered not only in the design of new inoculants in bioremediation but also in biodegradation assessments of chemicals present in natural environments

The main goal of this study wasto use an oleophilic biostimulant (S-200) to target possible nutritional limitations for biodegradation of polycyclic aromatic hydrocarbons (PAHs) at the interface between nonaqueous-phase liquids (NAPLs) and the water phase. Biodegradation of PAHs present in fuel-containing NAPLs was slow and followed zero-order kinetics, indicating bioavailability restrictions. The biostimulant enhanced the biodegradation, producing logistic (S-shaped) kinetics and 10fold increases in the rate of mineralization of phenanthrene, fluoranthene, and pyrene. Chemical analysis of residual fuel oil also evidenced an enhanced biodegradation of the alkyl- PAHs and n-alkanes. The enhancement was not the result of an increase in the rate of partitioning of PAHs into the aqueous phase, nor was it caused by the compensation of any nutritional deficiency in the medium. We suggest that biodegradation of PAH by bacteria attached to NAPLs can be limited by nutrient availability due to the simultaneous consumption of NAPL components, but this limitation can be overcome by interface fertilization.

Water quality degradation in harbour domains can have an important negative impact from an economic, touristic and environmental point of view. In that sense, water quality management is becoming a main concern for harbour managers. In this paper, we present the research behind the initiative started in Spanish harbours to control water quality degradation due to accidental pollution. This management system is already operationally running in the Barcelona harbour (NW Mediterranean Sea). The system is based on a recent published risk assessment, which takes into account not only the different activities in the harbour and their inherent risk of accident but also the physical behaviour of harbour waters. In this methodology, a key element is to get hydrodynamic forecasts. Thus, the system is composed of a hierarchy of nested hydrodynamic models covering from the basin scale to the harbour scale and a module that computes the different parameters needed for risk assessment. Special emphasis is made on describing the steps followed for system implementation because such implementation is far from a mere technical problem. The first step is to identify the main forcing factors for the harbour hydrodynamics from both field data and numerical experiments, which has never been done before for the Barcelona harbour. Wind and shelf currents are suggested as the main forcing factors for the harbour circulation. The second step is to identify the requirements that a numerical model must fulfil in order to properly solve the Barcelona harbour's hydrodynamics. A high resolution (<50. m) three dimensional model able to prognostically calculate temperature and salinity evolution; full air-sea coupling is needed as well. The third step is to investigate the best operational strategy. We have found that small errors in the initial density profiles are acceptable for surface current forecasts but not for deep circulation. A cold start must be avoided and a 72. h spin-up is recommende

This contribution presents a method for assessing the risk of water degradation in harbour domains. The method describes a normalized Index of Risk, ranging from 0 to 1, which determines the risk of water degradation due to a pollution event. A branch-decision scheme of decision-making theories was implemented in order to obtain this index. This method evaluates the cost of each decision as a function of the vulnerability, proximity and toxicity of the potential contaminant. A novel feature of this method is that the risk is defined by considering the physical behaviour of the harbour (i.e. water circulation patterns). Regions where water residence time is high are considered more vulnerable to pollutant releases. This method could be implemented from an operational perspective, in which case an oceanographic operational system could be used to obtain current forecasts which in turn would be used to forecast risk maps.

When incubated with a creosote-polycyclic aromatic hydrocarbons (PAHs) mixture, the pyrene-degrading strain Mycobacterium sp. AP1 acted on three- and four-ring components, causing the simultaneous depletion of 25% of the total PAHs in 30 days. The kinetics of disappearance of individual PAHs was consistent with differences in aqueous solubility. During the incubation, a number of acid metabolites indicative of distinctive reactions carried out by high-molecular-weight PAH-degrading mycobacteria accumulated in the medium. Most of these metabolites were dicarboxylic aromatic acids formed as a result of the utilization of growth substrates (phenanthrene, pyrene, or fluoranthene) by multibranched pathways including meta- and ortho-ring-cleavage reactions: phthalic acid, naphthalene-1,8-dicarboxylic acid, phenanthrene-4,5-dicarboxylic acid, diphenic acid, Z-9-carboxymethylenefluorene-1-carboxylic acid, and 6,6′-dihydroxy-2,2′-biphenyl dicarboxylic acid. Others were dead-end products resulting from cometabolic oxidations on nongrowth substrates (fluorene meta-cleavage product). These results contribute to the general knowledge of the biochemical processes that determine the fate of the individual components of PAH mixtures in polluted soils. The identification of the partially oxidized compounds will facilitate to develop analytical methods to determine their potential formation and accumulation in contaminated sites.