Replica Plating Method for Estimating Phenanthrene-Utilizing and Phenanthrene-Cometabolizing Microorganisms (original) (raw)
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The aim of this work was to develop a method to assess the microbial accessibility of native phenanthrene present in soils and sediments. We developed an accelerated biodegradation assay, characterized by (a) inoculation with a suYcient number of phenanthrene-degrading microorganisms, (b) monitoring of the biodegradation activity through 14 Cmineralization measurements, and (c) single-step chemical analysis of the native compound in the residue. The use of 14 C-labeling allowed the determination of the time period needed for biodegradation of the bioaccessible fraction of the native chemical. The method was tested with environmental samples having a wide range of phenanthrene concentrations, i.e., from background levels ( g kg -1 ) originating in soil from atmospheric deposition, to acute concentrations (g kg -1 ) corresponding to industrial pollution of soils and sediments. The results showed a wide range of bioaccessibility (15-95% of the initial amount). The method can be used for the assessment of bioaccessibility involved in the management of polycyclic aromatic hydrocarbon (PAH) pollution.
Applied and Environmental Microbiology, 1994
The relative role of eukaryotic versus prokaryotic microorganisms in phenanthrene transformation was measured in slurries of coastal sediment by two different approaches: detection of marker metabolites and use of selective inhibitors on phenanthrene biotransformation. Phenanthrene biotransformation was measured by polar metabolite formation and CO 2 evolution from [9- 14 C]phenanthrene. Radiolabeled metabolites were tentatively identified by high-performance liquid chromatography (HPLC) separation combined with UV/visible spectral analysis of HPLC peaks and comparison to authentic standards. Both yeasts and bacteria transformed phenanthrene in slurries of coastal sediment. Two products of phenanthrene oxidation by fungi, phenanthrene trans -3,4-dihydrodiol and 3-phenanthrol, were produced in yeast-inoculated sterile sediment. However, only products of phenanthrene oxidation typical of bacterial transformation, 1-hydroxy-2-naphthoic acid and phenanthrene cis -3,4-dihydrodiol, were i...
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Environmental Science and Pollution Research, 2014
Pollutants can severely impact structure and function of microbial communities. Biological interactions play a major role in structuring benthic food webs. We studied the impact of meiofauna on bacteria facing contamination by a mixture of 3 PAHs. Top down control by meiofauna was more effective than PAH in shaping bacterial diversity. The structural role of meiofauna disappeared when nutrients were added to enhance PAH biodegradation.
Marine Pollution Bulletin, 2002
Elevated concentrations of polycyclic aromatic hydrocarbons (PAHs) have been found in mangrove sediments due to anthropogenic pollution, and microbial degradation has been suggested as the best way to remove PAHs from contaminated sediments. The degradation of phenanthrene, a model PAH compound by bacteria, either the enriched mixed culture or individual isolate isolated from surface mangrove sediments was examined. The effects of salinity, initial phenanthrene concentrations and the addition of glucose on biodegradation potential were also investigated. Results show that surface sediments collected from four mangrove swamps in Hong Kong had different degree of PAH contamination and had different indigenous phenanthrene-degrading bacterial consortia. The enriched bacteria could use phenanthrene as the sole carbon source for growth and degrade this PAH compound accordingly. A significant positive relationship was found between bacterial growth and percentages of phenanthrene degradation. The phenanthrene biodegradation ability of the enriched mixed bacterial culture was not related to the degree of PAH contamination in surface sediments. The growth and biodegradation percentages of the enriched mixed culture were not higher than that of the individual isolate especially at low salinity (0 and 10 ppt). High salinity (35 ppt) inhibited growth and biodegradation of phenanthrene of a bacterial isolate but less inhibitory effect was found on the mixed culture. The inhibitory effects of salinity could be reduced with the addition of glucose.
The influence of sediment resuspension on the degradation of phenanthrene in flow-through microcosms
Marine Environmental Research, 2006
The effect of sediment resuspension on the mineralization of phenanthrene was examined in microcosms and sediment slurries. In computer-controlled, flow-through microcosms, 14C-phenanthrene-amended sediments were resuspended into overlying oxic water at frequencies of 12, 4, 1, 0.25 and 0 d−1. In slurry bottle experiments 14C-phenanthrene-amended sediments were continuously resuspended under oxic (excess air headspace) and anoxic (N2 headspace) conditions and mineralization was measured at periods from 2 h to 7 days. Our main findings were: (1) mineralization rate constants from the microcosms ranged from 0.001 to 0.01 d−1 and increased with frequency of resuspension, (2) these rates fell between those measured in oxic and anoxic slurries and were predicted within a factor of 2.5 by a model in which mineralization depended on the degree of oxygen exposure, and (3) the phenanthrene-degrading bacterial community was more active in resuspended sediments incubated in the microcosms than in sediments which were not resuspended, or which were stored under refrigeration. We conclude from these experiments that the effects of sediment resuspension on phenanthrene degradation are consistent with a primary role of average oxygen exposure, and also an alteration in the PAH-degrading activity of microbial populations.
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The microbial diversity and functioning around oceanic islands is poorly described, despite its importance for ecosystem homeostasis. Here, we aimed to verify the occurrence of microbe-driven phenanthrene co-oxidation in the seawater surrounding the Trindade Island (Brazil). We also used Next-Generation Sequencing to evaluate the effects of aliphatic and polycyclic aromatic hydrocarbons (PAHs) on these microbial community assemblies. Microcosms containing seawater from the island enriched with either labelled (9-(14)C) or non-labelled phenanthrene together with hexadecane, weathered oil, fluoranthene or pyrene, and combinations of these compounds were incubated. Biodegradation of phenanthrene-9-(14)C was negatively affected in the presence of weathered oil and PAHs but increased in the presence of hexadecane. PAH contamination caused shifts in the seawater microbial community-from a highly diverse one dominated by Alphaproteobacteria to less diverse communities dominated by Gammapro...