Endolithic Microbial Carbon Cycling in East Antarctica.

Antarctica is an ideal analogue for studying the limits of life. Despite severe temperature fluctuations and desiccating conditions, life is commonly found colonizing the structural cavities within Antarctic rocks (i.e., endoliths). Previous studies have speculated that the slow cycling of endoliths in the McMurdo Dry Valleys may be the limit of life on Earth. However, very little is known about the in situ activities of these communities-especially in regions outside the McMurdo Dry Valleys where endoliths are thought to be cycling carbon very slowly (e.g., hundreds of years). Here, we show that East Antarctic endoliths found on nunataks are cycling carbon quickly and are therefore quite active. Through radiocarbon (14C) analyses of the viable cell membrane (as phospholipid-derived fatty acids [PLFA]), we found that the Δ14C composition of these microbial communities was on average predominantly modern, with a few samples signaling older carbon in the system. These findings indicate that endoliths inhabiting inland Antarctic nunataks are cycling carbon on decadal timescales, which support the notion that endoliths in Antarctica are cycling carbon quickly. This work provides new insights into the potential variability of Antarctic endolith activities and demonstrates that, despite the climatic extremes that exist farther inland on the most inhospitable continent on Earth, indigenous life can thrive.

The Biodiversity and Geochemistry of Cryoconite Holes in Queen Maud Land, East Antarctica.

Cryoconite holes are oases of microbial diversity on ice surfaces. In contrast to the Arctic, where during the summer most cryoconite holes are 'open', in Continental Antarctica they are most often 'lidded' or completely frozen year-round. Thus, they represent ideal systems for the study of microbial community assemblies as well as carbon accumulation, since individual cryoconite holes can be isolated from external inputs for years. Here, we use high-throughput sequencing of the 16S and 18S rRNA genes to describe the bacterial and eukaryotic community compositions in cryoconite holes and surrounding lake, snow, soil and rock samples in Queen Maud Land. We cross correlate our findings with a broad range of geochemical data including for the first time 13C and 14C analyses of Antarctic cryoconites. We show that the geographic location has a larger effect on the distribution of the bacterial community compared to the eukaryotic community. Cryoconite holes are distinct from the local soils in both 13C and 14C and their isotopic composition is different from similar samples from the Arctic. Carbon contents were generally low (≤0.2%) and older (6-10 ky) than the surrounding soils, suggesting that the cryoconite holes are much more isolated from the atmosphere than the soils.

Ongoing biodegradation of Deepwater Horizon oil in beach sands: Insights from tracing petroleum carbon into microbial biomass.

Heavily weathered petroleum residues from the Deepwater Horizon (DwH) disaster continue to be found on beaches along the Gulf of Mexico as oiled-sand patties. Here, we demonstrate the ongoing biodegradation of weathered Macondo Well (MW) oil residues by tracing oil-derived carbon into active microbial biomass using natural abundance radiocarbon (14C). Oiled-sand patties and non-oiled sand were collected from previously studied beaches in Mississippi, Alabama, and Florida. Phospholipid fatty acid (PLFA) analyses illustrated that microbial communities present in oiled-sand patties were distinct from non-oiled sand. Depleted 14C measurements of PLFA revealed that microbes on oiled-sand patties were assimilating MW oil residues five years post-spill. In contrast, microbes in non-oiled sand assimilated recently photosynthesized carbon. These results demonstrate ongoing biodegradation of weathered oil in sand patties and the utility of 14C PLFA analysis to track the biodegradation of MW oil residues long after other indicators of biodegradation are no longer detectable.

Elucidating carbon sources driving microbial metabolism during oil sands reclamation.

Microbial communities play key roles in remediation and reclamation of contaminated environments via biogeochemical cycling of organic and inorganic components. Understanding the trends in in situ microbial community abundance, metabolism and carbon sources is therefore a crucial component of effective site management. The focus of this study was to use radiocarbon analysis to elucidate the carbon sources driving microbial metabolism within the first pilot wetland reclamation project in the Alberta oil sands region where the observation of H2S had indicated the occurrence of microbial sulphate reduction. The reclamation project involved construction of a three compartment system consisting of a freshwater wetland on top of a sand cap overlying a composite tailings (CT) deposit. Radiocarbon analysis demonstrated that both dissolved and sediment associated organic carbon associated with the deepest compartments (the CT and sand cap) was primarily fossil (Δ14C = -769 to -955‰) while organic carbon in the overlying peat was hundreds to thousands of years old (Δ14C = -250 to -350‰). Radiocarbon contents of sediment associated microbial phospholipid fatty acids (PLFA) were consistent with the sediment bulk organic carbon pools (Peat: Δ14CPLFA = -257‰; Sand cap Δ14CPLFA = -805‰) indicating that these microbes were using sediment associated carbon. In contrast, microbial PLFA grown on biofilm units installed in wells within the deepest compartments contained much more modern carbon that the associated bulk carbon pools. This implied that the transfer of relatively more modern carbon was stimulating the microbial community at depth within the system. Correlation between cellular abundance estimates based on PLFA concentrations and the Δ14CPLFA indicated that the utilization of this more modern carbon was stimulating the microbial community at depth. These results highlight the importance of understanding the occurrence and potential outcomes of the introduction of relatively bioavailable carbon to mine wastes in order to predict and manage the performance of reclamation strategies.

Radiocarbon evidence of active endolithic microbial communities in the hyperarid core of the Atacama Desert.

The hyperarid core of the Atacama Desert is one of the driest and most inhospitable places on Earth, where life is most commonly found in the interior of rocks (i.e., endolithic habitats). Due to the extreme dryness, microbial activity in these habitats is expected to be low; however, the rate of carbon cycling within these microbial communities remains unknown. We address this issue by characterizing the isotopic composition ((13)C and (14)C) of phospholipid fatty acids (PLFA) and glycolipid fatty acids (GLFA) in colonized rocks from four different sites inside the hyperarid core. δ(13)C results suggest that autotrophy and/or quantitative conversion of organic matter to CO2 are the dominant processes occurring with the rock. Most Δ(14)C signatures of PLFA and GLFA were consistent with modern atmospheric CO2, indicating that endoliths are using atmospheric carbon as a primary carbon source and are also cycling carbon quickly. However, at one site the PLFA contained (14)C from atmospheric nuclear weapons testing that occurred during the 1950s and 1960s, indicating a decadal rate of carbon cycling. At the driest site (Yungay), based on the relative abundance and (14)C content of GLFA and PLFA, there was evidence of possible preservation. Hence, in low-moisture conditions, glycolipids may persist while phospholipids are preferentially hydrolyzed.

A carbon free filter for collection of large volume samples of cellular biomass from oligotrophic waters.

Isotopic analysis of cellular biomass has greatly improved our understanding of carbon cycling in the environment. Compound specific radiocarbon analysis (CSRA) of cellular biomass is being increasingly applied in a number of fields. However, it is often difficult to collect sufficient cellular biomass for analysis from oligotrophic waters because easy-to-use filtering methods that are free of carbon contaminants do not exist. The goal of this work was to develop a new column based filter to autonomously collect high volume samples of biomass from oligotrophic waters for CSRA using material that can be baked at 450°C to remove potential organic contaminants. A series of filter materials were tested, including uncoated sand, ferrihydrite-coated sand, goethite-coated sand, aluminum-coated sand, uncoated glass wool, ferrihydrite-coated glass wool, and aluminum-coated glass wool, in the lab with 0.1 and 1.0 µm microspheres and Escherichia coli. Results indicated that aluminum-coated glass wool was the most efficient filter and that the retention capacity of the filter far exceeded the biomass requirements for CSRA. Results from laboratory tests indicate that for oligotrophic waters with 1×10(5) cells ml(-1), 117l of water would need to be filtered to collect 100 µg of PLFA for bulk PLFA analysis and 2000 l for analysis of individual PLFAs. For field sampling, filtration tests on South African mine water indicated that after filtering 5955l, 450 µg of total PLFAs were present, ample biomass for radiocarbon analysis. In summary, we have developed a filter that is easy to use and deploy for collection of biomass for CSRA including total and individual PLFAs.

Quantification of extraneous carbon during compound specific radiocarbon analysis of black carbon.

Radiocarbon ((14)C) is a radioactive isotope that is useful for determining the age and cycling of carbon-based materials in the Earth system. Compound specific radiocarbon analysis (CSRA) provides powerful insight into the turnover of individual components that make up the carbon cycle. Extraneous or nonspecific background carbon (C(ex)) is added during sample processing and subsequent isolation of CSRA samples. Here, we evaluate the quantity and radiocarbon signature of C(ex) added from two sources: preparative capillary gas chromatography (PCGC, C(PCGC)) and chemical preparation of CSRA of black carbon samples (C(chemistry)). We evaluated the blank directly using process blanks and indirectly by quantifying the difference in the isotopic composition between processed and unprocessed samples for a range of sample sizes. The direct and indirect assessment of C(chemistry+PCGC) agree, both in magnitude and radiocarbon value (1.1 +/- 0.5 microg of C, fraction modern = 0.2). Half of the C(ex) is introduced before PCGC isolation, likely from coeluting compounds in solvents used in the extraction method. The magnitude of propagated uncertainties of CSRA samples are a function of sample size and collection duration. Small samples collected for a brief amount of time have a smaller propagated (14)C uncertainty than larger samples collected for a longer period of time. CSRA users are cautioned to consider the magnitude of uncertainty they require for their system of interest, to frequently evaluate the magnitude of C(ex) added during sampling processing, and to avoid isolating samples < or = 5 microg of carbon.

The feasibility of isolation and detection of fullerenes and carbon nanotubes using the benzene polycarboxylic acid method.

The incorporation of fullerenes and carbon nanotubes into electronic, optical and consumer products will inevitably lead to the presence of these anthropogenic compounds in the environment. To date, there have been few studies isolating these materials from environmental matrices. Here we report a method commonly used to quantify black carbon (BC) in soils, the benzene polycarboxylic acid (BPCA) method, for measurement of two types of single walled carbon nanotubes (SWCNTs), two types of fullerenes and two forms of soot. The distribution of BC products (BPCAs) from the high pressure and high temperature oxidation illustrates the condensed nature of these compounds because they form predominantly fully substituted mellitic acid (B6CA). The conversion of carbon nanoparticles to BPCAs was highest for fullerenes (average of 23.2+/-4.0% C recovered for both C(60) and C(70)) and lowest for non-functionalized SWCNTs (0.5+/-0.1% C). The recovery of SWCNTs was 10 times higher when processed through a cation-exchange column, indicating the presence of metals in SWCNTs compromises the oxidation chemistry. While mixtures of SWCNTs, soot and sediment revealed small losses of black carbon during sample processing, the method is suitable for quantifying total BC. The BPCA distribution of mixtures did not agree with theoretical mixtures using model polyaromatic hydrocarbons, suggesting the presence of a matrix effect. Future work is required to quantify different types of black carbon within the same sample.