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Aims: Our goal was to characterize a newly isolated strain of Mycobacterium austroafricanum, obtained from manufactured gas plant (MGP) site soil and designated GTI-23, with respect to its ability to degrade polycyclic aromatic hydrocarbons (PAHs). Methods and Results: GTI-23 is capable of growth on phenanthrene, fluoranthene, or pyrene as a sole source of carbon and energy; it also extensively mineralizes the latter two in liquid culture and is capable of extensive degradation of fluorene and benzo[a]pyrene, although this does not lead in either of these cases to mineralization. Supplementation of benzo [a]pyrene-containing cultures with phenanthrene had no significant effect on benzo[a]pyrene degradation; however, this process was substantially inhibited by the addition of pyrene. Extensive and rapid mineralization of pyrene by GTI-23 was also observed in pyrene-amended soil. Conclusions: Strain GTI-23 shows considerable ability to mineralize a range of polycyclic aromatic hydrocarbons, both in liquid and soil environments. In this regard, GTI-23 differs markedly from the type strain of Myco. austroafricanum (ATCC 33464); the latter isolate displayed no (or very limited) mineralization of any tested PAH (phenanthrene, fluoranthene or pyrene). When grown in liquid culture, GTI-23 was also found to be capable of growing on and mineralizing two aliphatic hydrocarbons (dodecane and hexadecane). Significance and Impact of the Study: These findings indicate that this isolate of Myco. austroafricanum may be useful for bioremediation of soils contaminated with complex mixtures of aromatic and aliphatic hydrocarbons.

Alkanindiges illinoisensis gen. nov., sp. nov., an obligately hydrocarbonoclastic, aerobic squalanedegrading bacterium isolated from oilfield soils An alkane-degrading bacterium, designated GTI MVAB Hex1 T , was isolated from chronically crude oil-contaminated soil from an oilfield in southern Illinois. The isolate grew very weakly or not at all in minimal or rich media without hydrocarbons. Straight-chain aliphatic hydrocarbons, such as hexadecane and heptadecane, greatly stimulated growth; shorter-chain (¡C 15 ) hydrocarbons did not (with decane as the sole exception). Growth was also greatly enhanced by the branched aliphatic hydrocarbons pristane and squalane. The latter of these was most intriguing, as catabolism of squalane has hitherto been reported only for Mycobacterium species. Although unable to utilize mono-or polycyclic aromatic hydrocarbons as sole carbon sources, the isolate did show slight fluorene-mineralizing capability in Luria-Bertani medium, which was partially repressed by hexadecane. In contrast, hexadecane supplementation greatly increased mineralization of 14 C-dodecane, which was not a growth substrate. Further testing emphasized the isolate's extremely narrow substrate range, as only Tween 40 and Tween 80 supported significant growth. Microscopic examination (by scanning and transmission electron microscopy) revealed a slightly polymorphic coccoidal to bacillar morphology, with hydrocarbon-grown cells tending to be more elongated. When grown with hexadecane, GTI MVAB Hex1 T accumulated a large number of electron-transparent intracytoplasmic inclusion bodies. These were also prevalent during growth in the presence of squalane. Smaller inclusion bodies were observed occasionally with pristane supplementation; they were, however, absent during growth on crude oil. On the basis of 16S rRNA gene sequence data and range of growth substrates, classification of this isolate as the type strain of Alkanindiges illinoisensis gen. nov., sp. nov. is proposed, which is most closely related (approx. 94 % sequence similarity) to Acinetobacter junii.Bacterial biodegradation of crude oil first gained widespread attention as a potential remediation approach in the 1970s (Atlas & Bartha, 1972;Soli & Bens, 1972). Many bacteria degrade aliphatic constituents of crude oils, including straight-chain aliphatic hydrocarbons up to C 44 (Sakai et al., 1994;Radwan et al., 1996). Branched-chain alkanes, particularly those with anteiso-terminal branching [substitution on carbon atoms that are immediately adjacent to the terminal carbon atom(s)], are much less susceptible to b-oxidation (Schaeffer et al., 1979), and thus more recalcitrant, than their unbranched counterparts. Thus, pristane (2,6,10,14-tetramethylpentadecane) was often used as an internal standard to determine biodegradation and weathering of other oil fractions (Alvarez et al., 2001). More recently, however, species of several genera have been shown to biodegrade pristane (McKenna & Kallio, 1971;Pirnik et al., 1974;Nakajima & Sato, 1983;Lal & Khanna, ...

We report development of a novel parameter for quantifying the amount of humic and fulvic acids per unit surface area in a particular soil. This quantity, the "humic coverage index" (HCI), provides a measurement of the relative spatial extents and/or thicknesses of the humic/fulvic overlayers in different soils, and, therefore, can be used in modeling various soils' behavior in sequestration processes in which humic materials are involved. HCI is herein applied to modeling biodegradation of aromatic and aliphatic hydrocarbons (phenanthrene, pyrene, and hexadecane) by several bacterial strains. Results indicate that, for the cases studies here, contaminant biodegradation is highest at a particular HCI and decreases if the coverage density of humic material is lower or higher than this optimum value. The HCI value at which maximal degradation was observed varied across different strains (indicating strain-specific differences in ability to degrade contaminants sorbed to humic materials) and, to a lesser extent, across different contaminants. The HCI concept is also demonstrated to be useful in explaining soil-, strain-, and contaminant-specific variations in the ability of fulvic acid supplementation to enhance contaminant biodegradation. Finally, we show that, in general, strains which are comparatively better at degrading contaminants in high-HCI soils also show enhanced contaminant mineralization in vitro in the presence of humic acids, such as when hydrocarbons are adsorbed onto these materials.

Studies were conducted to examine the mobility and bioavailability to earthworms (Eisenia fetida) of priority pollutant polycyclic aromatic hydrocarbons (PAH) in a suite of 11 soils and soil/lampblack mixtures obtained from former manufactured-gas plant sites. Contaminant mobility was assessed using XAD4 resins encapsulated in dialysis tubing, which were exposed to slurried soils for 15 d. These experiments showed that mobility of PAH in the different soils strongly correlated to the levels of volatile hydrocarbons (namely, gasoline- and diesel-range organics [GRO and DRO]) that existed in the soils as co-contaminants. Actual PAH bioavailability (as measured by earthworm PAH concentrations) also appeared to depend on GRO + DRO levels, although this was most evident at high levels of these contaminants. These findings are discussed in view of the effects of dieselrange organics on oil viscosity, assuming that the hydrocarbon contaminants in these soils exist in the form of distinct adsorbed oil phases. This study, therefore, extends correlations between carrier-oil viscosity and dissolved solute bioavailability, previously observed in a number of other in vitro and whole-organism tests (and in bacterial mutagenicity studies in soil), to multicellular organisms inhabiting contaminated-soil systems.