Molecular diversity of a North Carolina wastewater treatment plant as revealed by pyrosequencing.

We report the results of pyrosequencing of DNA collected from the activated sludge basin of a wastewater treatment plant in Charlotte, NC. Using the 454-FLX technology, we generated 378,601 sequences with an average read length of 250.4 bp. Running the 454 assembly algorithm over our sequences yielded very poor assembly, with only 0.3% of our sequences participating in assembly of significant contigs. Of the 117 contigs greater than 500 bp long that were assembled, the most common annotations were to transposases and hypothetical proteins. Comparing our sequences to known microbial genomes showed nonspecific recruitment, indicating that previously described taxa are only distantly related to the most abundant microbes in this treatment plant. A comparison of proteins generated by translating our sequence set to translations of other sequenced microbiomes shows a distinct metabolic profile for activated sludge with high counts for genes involved in metabolism of aromatic compounds and low counts for genes involved in photosynthesis. Taken together, these data document the substantial levels of microbial diversity within activated sludge and further establish the great utility of pyrosequencing for investigating diversity in complex ecosystems.

Bio-tarp alternative daily cover prototypes for methane oxidation atop open landfill cells.

Final landfill covers are highly engineered to prevent methane release into the atmosphere. However, methane production begins soon after waste placement and is an unaddressed source of emissions. The methane oxidation capacity of methanotrophs embedded in a "bio-tarp" was investigated as a means to mitigate methane release from open landfill cells. The bio-tarp would also serve as an alternative daily cover during routine landfill operation. Evaluations of nine synthetic geotextiles identified two that would likely be suitable bio-tarp components. Pilot tarp prototypes were tested in continuous flow systems simulating landfill gas conditions. Multilayered bio-tarp prototypes consisting of alternating layers of the two geotextiles were found to remove 16% of the methane flowing through the bio-tarp. The addition of landfill cover soil, compost, or shale amendments to the bio-tarp increased the methane removal up to 32%. With evidence of methane removal in a laboratory bioreactor, prototypes were evaluated at a local landfill using flux chambers installed atop intermediate cover at a landfill. The multilayered bio-tarp and amended bio-tarp configurations were all found to decrease landfill methane flux; however, the performance efficacy of bio-tarps was not significantly different from controls without methanotrophs. Because highly variable methane fluxes at the field site likely confounded the test results, repeat field testing is recommended under more controlled flux conditions.

Microbial methane oxidation processes and technologies for mitigation of landfill gas emissions.

Landfill gas containing methane is produced by anaerobic degradation of organic waste. Methane is a strong greenhouse gas and landfills are one of the major anthropogenic sources of atmospheric methane. Landfill methane may be oxidized by methanotrophic microorganisms in soils or waste materials utilizing oxygen that diffuses into the cover layer from the atmosphere. The methane oxidation process, which is governed by several environmental factors, can be exploited in engineered systems developed for methane emission mitigation. Mathematical models that account for methane oxidation can be used to predict methane emissions from landfills. Additional research and technology development is needed before methane mitigation technologies utilizing microbial methane oxidation processes can become commercially viable and widely deployed.

Biotic systems to mitigate landfill methane emissions.

Landfill gases produced during biological degradation of buried organic wastes include methane, which when released to the atmosphere, can contribute to global climate change. Increasing use of gas collection systems has reduced the risk of escaping methane emissions entering the atmosphere, but gas capture is not 100% efficient, and further, there are still many instances when gas collection systems are not used. Biotic methane mitigation systems exploit the propensity of some naturally occurring bacteria to oxidize methane. By providing optimum conditions for microbial habitation and efficiently routing landfill gases to where they are cultivated, a number of bio-based systems, such as interim or long-term biocovers, passively or actively vented biofilters, biowindows and daily-used biotarps, have been developed that can alone, or with gas collection, mitigate landfill methane emissions. This paper reviews the science that guides bio-based designs; summarizes experiences with the diverse natural or engineered substrates used in such systems; describes some of the studies and field trials being used to evaluate them; and discusses how they can be used for better landfill operation, capping, and aftercare.