Search

Summary Mycobacterium is often isolated from polycyclic aromatic hydrocarbon (PAH)-contaminated soil as degraders of PAHs. In model systems, Mycobacterium shows attachment to the PAH substrate source, which is considered to be a particular adaptation to low bioavailability as it results into increased substrate flux to the degraders. To examine whether PAH-degrading Mycobacterium in real PAH-contaminated soils, in analogy with model systems, are preferentially associated with PAH-enriched soil particles, the distribution of PAHs, of the PAH-mineralizing capacity and of Mycobacterium over different fractions of a soil with an aged PAH contamination was investigated. The clay fraction contained the majority of the PAHs and showed immediate pyrene- and phenanthrene-mineralizing activity upon addition of (14)C-labelled pyrene or phenanthrene. In contrast, the sand and silt fractions showed a lag time of 15-26 h for phenanthrene and 3-6 days for pyrene mineralization. The maximum pyrene and phenanthrene mineralization rates of the clay fraction expressed per gram fraction were three to six times higher than those of the sand and silt fractions. Most-probable-number (MPN)-polymerase chain reaction demonstrated that Mycobacterium represented about 10% of the eubacteria in the clay fraction, while this was only about 0.1% in the sand and silt fractions, indicating accumulation of Mycobacterium in the PAH-enriched clay fraction. The Mycobacterium community composition in the clay fraction represented all dominant Mycobacterium populations of the bulk soil and included especially species related to Mycobacterium pyrenivorans, which was also recovered as one of the dominant species in the eubacterial communities of the bulk soil and the clay fraction. Moreover, Mycobacterium could be identified among the major culturable PAH-degrading populations in both the bulk soil and the clay fraction. The results demonstrate that PAH-degrading mycobacteria are mainly associated with the PAH-enriched clay fraction of the examined PAH-contaminated soil and hence, that also in the environmental setting of a PAH-contaminated soil, Mycobacterium might experience advantages connected to substrate source attachment.

The bioavailability to bacteria of 14C-labeled polycyclic aromatic hydrocarbons (PAHs) sorbed onto lake sediments was assessed using a mathematical model and three experimental series. The experiments were performed under similar conditions and included: (1) abiotic desorption of PAHs from sediments by Tenax extraction, (2) mineralization of dissolved PAHs with no sediment present, and (3) mineralization of PAHs sorbed onto sediments. Results obtained from the first two series were used to obtain the parameter values for the model, and the experimental results of the third series were compared to model results. We found that microorganisms were able to promote desorption of the more-labile fractions, but were unable to increase the desorption rate of the slow- and very slow-desorbing fractions. Also, our model predictions indicate that, after very long contact times, and in the concurrence of biodegradation, sorbed PAHs remain not under equilibrium conditions, but rather in a steady state. The net rates of PAH desorption from the three sediment domains considered (fast, slow, and very slow) become similar, and the ratio between the aqueous and the sediment concentration remains constant with time.

A study was conducted on the effect of two different biological factors, microbial surfactants and biodegradation, on the kinetics of partitioning of polycyclic aromatic hydrocarbons (PAHs) from nonaqueous-phase liquids (NAPLs). The effect of rhamnolipid biosurfactants on partitioning into the aqueous phase of naphthalene, fluorene, phenanthrene, and pyrene, initially dissolved in di-2-ethylhexyl phthalate (DEHP) or 2,2,4,4,6,8,8-heptamethylnonane (HMN), was determined in multiple-solute experiments. Biosurfactants at a concentration above the CMC enhanced the partitioning rate of fluorene, phenanthrene, and pyrene but were ineffective with naphthalene. Enhancement of partitioning was also observed in the presence of suspended humic acid-clay complexes, which simulated the solids often present in the subsurface. Biosurfactants sorbed to the complexes modified PAH partitioning between the NAPL and these solids, increasing the fraction of solid-phase PAH. Biodegradation-driven partitioning was estimated in mineralization experiments with phenanthrene initially present in HMN and three representative soil bacterial strains, differing in their potential adherence to the NAPL. In the three cases, the rates of mineralization were very similar and significantly higher than the abiotic rate of partitioning. Our study suggests that in NAPL-polluted sites, partitioning of PAH may be efficiently enhanced by in situ treatments involving the use of biosurfactants and biodegradation.

Abstract The limited mass transfer in polycyclic aromatic hydrocarbon (PAH)-contaminated soils during bioremediation treatments often impedes the achievement of regulatory decontamination end-points. Little is known about bioavailability of these hydrophobic pollutants in phytoremediation systems. This work attempts to evaluate, for the first time, chemotaxis as a bioavailability-promoting trait in PAH-degrading bacteria from the rhizosphere. For this aim, 20 motile strains capable of degrading different PAHs were isolated from rhizosphere soils contaminated with coal tar and oil. Three representative Pseudomonas strains were selected, on the basis of their faster growth and/or range of PAHs degraded, for detailed chemotaxis studies with PAHs (naphthalene, phenanthrene, anthracene, and pyrene), bacterial lipopolysaccharide and root exudates from seven different plants. The chemotactic response was quantified with a new densitometric method. The results indicate that chemotaxis is a relevant mobilizing factor for PAH-degrading rhizosphere bacteria.

Abstract Methods that quickly assess microbial density and aggregation in soil and sediments are needed in environmental microbiology. We report a flow cytometry method that uses the green and orange emission of the fluorochrome SYTO-13 to discriminate between bacteria and clay-humic acid particles. This approach distinguishes single or clustered bacteria, and clusters of bacteria and abiotic particles during the growth of the biosurfactant-producing strain Pseudomonas aeruginosa 19SJ on solid phenanthrene in the presence of humic acid-clay complexes.

The biodegradation of phenanthrene by the biosurfactant-producing strain Pseudomonas aeruginosa 19SJ was investigated in experiments with the compound present either as crystals or dissolved in non-aqueous phase liquids (NAPLs). Growth on solid phenanthrene exhibited an initial phase not limited by dissolution rate and a subsequent, carbon-limited phase caused by exhaustion of the carbon source. Rhamnolipid biosurfactants were produced from solid phenanthrene and appeared in solution and particulate material (cells and phenanthrene crystals). During the carbon-limited phase, the concentration of rhamnolipids detected in culture exceeded the critical micelle concentration (CMC) determined with purified rhamnolipids. The biosurfactants caused a significant increase in dissolution rate and pseudosolubility of phenanthrene, but only at concentrations above the CMC. Externally added rhamnolipids at a concentration higher than the CMC increased the biodegradation rate of solid phenanthrene. Mineralization curves of low concentrations of phenanthrene initially dissolved in two NAPLs [2,2,4,4,6,8,8-heptamethylnonane and di(2-ethylhexyl)phthalate] were S-shaped, although no growth was observed in the population of suspended bacteria. Biosurfactants were not detected in solution under these conditions. The observed mineralization was attributed not only to suspended bacteria, but also to bacterial populations growing at the NAPL-water interface, mineralizing the compound at higher rates than predicted by abiotic partitioning. We suggest that rhamnolipid production and attachment increased the bioavailability of phenanthrene, so promoting biodegradation activity.

The mineralization of phenanthrene in pure cultures of a Pseudomonas fluorescens strain, isolated from soil, was measured in the presence of soil humic fractions and montmorillonite. Humic acid and clay, either separately or in combination, shortened the acclimation phase. A higher mineralization rate was measured in treatments with humic acid at 100 microg/ml. Humic acid at 10 microg/ml stimulated the transformation only in the presence of 10 g of clay per liter. We suggest that sorption of phenanthrene to these soil components may result in a higher concentration of substrate in the vicinity of the bacterial cells and therefore may increase its bioavailability.

The mineralization by an Arthrobacter sp. of naphthalene initially dissolved in di(2-ethylhexyl)phthalate exhibited a slow phase followed by a rapid phase. Triton X-100, which inhibited cell attachment, prevented the onset of the second phase. Triton X-100 increased the extent of mineralization of naphthalene initially present in 2,2,4,4,6,8,8-heptamethylnonane. Cells attached to the interface mineralized the aromatic hydrocarbon at a rate four times higher than the rate of partitioning in the absence of microorganisms, but this microbial activity was markedly reduced by Triton X-100. We suggest that utilization of naphthalene originally present in nonaqueous-phase liquids may involve a partitioning-limited initial stage carried out by bacteria freely suspended in the aqueous phase and a subsequent, more rapid stage effected by bacteria present directly at the nonaqueous-liquid-water interface.