Enrichment, isolation, and phylogenetic identification of polycyclic aromatic hydrocarbon-degrading bacteria from Elizabeth River sediments.

The diversity of indigenous bacteria in sediments from several sites in the Elizabeth River (Virginia) able to degrade multiple polycyclic aromatic hydrocarbons (PAHs) was investigated by the use of classical selective enrichment and molecular analyses. Enrichment cultures containing naphthalene, phenanthrene, fluoranthene, or pyrene as a sole carbon and energy source were monitored by denaturing gradient gel electrophoresis (DGGE) to detect changes in the bacterial-community profile during enrichment and to determine whether the representative strains present were successfully cultured. The DGGE profiles of the final enrichments grown solely on naphthalene and pyrene showed no clear relationship with the site from which the inoculum was obtained. The enrichments grown solely on pyrene for two sample sites had >80% similarity, which suggests that common pyrene-degrading strains may be present in these sediments. The final enrichments grown on fluoranthene and phenanthrene remained diverse by site, suggesting that these strains may be influenced by environmental conditions. One hundred and one isolates were obtained, comprising representatives of the actinomycetes and alpha-, beta-, and gammaproteobacteria, including seven novel isolates with 16S rRNA gene sequences less than 98% similar to known strains. The ability to degrade multiple PAHs was demonstrated by mineralization of 14C-labeled substrate and growth in pure culture. This supports our hypothesis that a high diversity of bacterial strains with the ability to degrade multiple PAHs can be confirmed by the combined use of classical selective enrichment and molecular analyses. This large collection of diverse PAH-degrading strains provides a valuable resource for studies on mechanisms of PAH degradation and bioremediation.

High power density from a miniature microbial fuel cell using Shewanella oneidensis DSP10.

A miniature microbial fuel cell (mini-MFC) is described that demonstrates high output power per device cross-section (2.0 cm2) and volume (1.2 cm3). Shewanella oneidensis DSP10 in growth medium with lactate and buffered ferricyanide solutions were used as the anolyte and catholyte, respectively. Maximum power densities of 24 and 10 mW/m2 were measured using the true surface areas of reticulated vitreous carbon (RVC) and graphite felt (GF) electrodes without the addition of exogenous mediators in the anolyte. Current densities at maximum power were measured as 44 and 20 mA/m2 for RVC and GF, while short circuit current densities reached 32 mA/m2 for GF anodes and 100 mA/m2 for RVC. When the power density for GF was calculated using the cross sectional area of the device or the volume of the anode chamber, we found values (3 W/m2, 500 W/m3) similar to the maxima reported in the literature. The addition of electron mediators resulted in current and power increases of 30-100%. These power densities were surprisingly high considering a pure S. oneidensis culture was used. We found that the short diffusion lengths and high surface-area-to-chamber volume ratio utilized in the mini-MFC enhanced power density when compared to output from similar macroscopic MFCs.

The concentrations of hexadecane and inorganic nutrients modulate the production of extracellular membrane-bound vesicles, soluble protein, and bioemulsifier by Acinetobacter venetianus RAG-1 and Acinetobacter sp. strain HO1-N.

In the present study, we addressed the possibility that the production of both bioemulsifiers and membrane-bound vesicles may be a common feature of the growth of Acinetobacter spp. on alkanes, and we determined the extent to which the release of extracellular products by these organisms is regulated by the concentrations of the alkane substrate and inorganic nutrients. To accomplish this objective, we grew Acinetobacter venetianus RAG-1 and Acinetobacter sp. strain HO1-N with different concentrations of nutrients and assayed for extracellular products. The results indicated that the release of vesicles, soluble protein, and bioemulsifier was promoted in various degrees by higher concentrations of hexadecane and inorganic nutrients, while the specific activities of the bioemulsifiers were enhanced with lower nutrient concentrations. Based on our findings, we propose that under conditions of nutrient excess, these strains produce membrane-bound vesicles to function in "luxury uptake" of the alkane substrate for delivery and storage in the form of inclusions. Under the same conditions, soluble bioemulsifier and protein may perform auxiliary roles in cell desorption and (or) alkane uptake. With low concentrations of nutrients, the decreased production of vesicles, protein, and bioemulsifier and the increased activity of the emulsifier may represent a mechanism for reducing biosynthetic demands and conserving cellular material.