Biogeography and Photosynthetic Biomass of Arctic Marine Pico-Eukaroytes during Summer of the Record Sea Ice Minimum 2012

PubMed Central

Metfies, Katja; von Appen, Wilken-Jon; Kilias, Estelle; Nicolaus, Anja; Nöthig, Eva-Maria

2016-01-01

Information on recent photosynthetic biomass distribution and biogeography of Arctic marine pico-eukaryotes (0.2–3 μm) is needed to better understand consequences of environmental change for Arctic marine ecosystems. We analysed pico-eukaryote biomass and community composition in Fram Strait and large parts of the Central Arctic Ocean (Nansen Basin, Amundsen Basin) using chlorophyll a (Chl a) measurements, automated ribosomal intergenic spacer analysis (ARISA) and 454-pyrosequencing. Samples were collected during summer 2012, the year with the most recent record sea ice minimum. Chl a concentrations were highest in eastern Fram Strait and pico-plankton accounted for 60–90% of Chl a biomass during the observation period. ARISA-patterns and 454-pyrosequencing revealed that pico-eukaryote distribution is closely related to water mass distribution in the euphotic zone of the Arctic Ocean. Phaeocystaceae, Micromonas sp., Dinophyceae and Syndiniales constitute a high proportion of sequence reads, while sequence abundance of autotrophic Phaeocystaceae and mixotrophic Micromonas sp. was inversely correlated. Highest sequence abundances of Phaeocystaceae were observed in the warm Atlantic Waters in Fram Strait, while Micromonas sp. dominated the abundant biosphere in the arctic halocline. Our results are of particular interest considering existing hypotheses that environmental conditions in Nansen Basin might become more similar to the current conditions in Fram Strait. We propose that in response, biodiversity and biomass of pico-eukaryotes in Nansen Basin could resemble those currently observed in Fram Strait in the future. This would significantly alter biogeochemical cycles in a large part of the Central Arctic Ocean. PMID:26895333

Abiotic factors shape microbial diversity in Sonoran Desert soils.

PubMed

Andrew, David R; Fitak, Robert R; Munguia-Vega, Adrian; Racolta, Adriana; Martinson, Vincent G; Dontsova, Katerina

2012-11-01

High-throughput, culture-independent surveys of bacterial and archaeal communities in soil have illuminated the importance of both edaphic and biotic influences on microbial diversity, yet few studies compare the relative importance of these factors. Here, we employ multiplexed pyrosequencing of the 16S rRNA gene to examine soil- and cactus-associated rhizosphere microbial communities of the Sonoran Desert and the artificial desert biome of the Biosphere2 research facility. The results of our replicate sampling approach show that microbial communities are shaped primarily by soil characteristics associated with geographic locations, while rhizosphere associations are secondary factors. We found little difference between rhizosphere communities of the ecologically similar saguaro (Carnegiea gigantea) and cardón (Pachycereus pringlei) cacti. Both rhizosphere and soil communities were dominated by the disproportionately abundant Crenarchaeota class Thermoprotei, which comprised 18.7% of 183,320 total pyrosequencing reads from a comparatively small number (1,337 or 3.7%) of the 36,162 total operational taxonomic units (OTUs). OTUs common to both soil and rhizosphere samples comprised the bulk of raw sequence reads, suggesting that the shared community of soil and rhizosphere microbes constitute common and abundant taxa, particularly in the bacterial phyla Proteobacteria, Actinobacteria, Planctomycetes, Firmicutes, Bacteroidetes, Chloroflexi, and Acidobacteria. The vast majority of OTUs, however, were rare and unique to either soil or rhizosphere communities and differed among locations dozens of kilometers apart. Several soil properties, particularly soil pH and carbon content, were significantly correlated with community diversity measurements. Our results highlight the importance of culture-independent approaches in surveying microbial communities of extreme environments.

Abiotic Factors Shape Microbial Diversity in Sonoran Desert Soils

PubMed Central

Fitak, Robert R.; Munguia-Vega, Adrian; Racolta, Adriana; Martinson, Vincent G.; Dontsova, Katerina

2012-01-01

High-throughput, culture-independent surveys of bacterial and archaeal communities in soil have illuminated the importance of both edaphic and biotic influences on microbial diversity, yet few studies compare the relative importance of these factors. Here, we employ multiplexed pyrosequencing of the 16S rRNA gene to examine soil- and cactus-associated rhizosphere microbial communities of the Sonoran Desert and the artificial desert biome of the Biosphere2 research facility. The results of our replicate sampling approach show that microbial communities are shaped primarily by soil characteristics associated with geographic locations, while rhizosphere associations are secondary factors. We found little difference between rhizosphere communities of the ecologically similar saguaro (Carnegiea gigantea) and cardón (Pachycereus pringlei) cacti. Both rhizosphere and soil communities were dominated by the disproportionately abundant Crenarchaeota class Thermoprotei, which comprised 18.7% of 183,320 total pyrosequencing reads from a comparatively small number (1,337 or 3.7%) of the 36,162 total operational taxonomic units (OTUs). OTUs common to both soil and rhizosphere samples comprised the bulk of raw sequence reads, suggesting that the shared community of soil and rhizosphere microbes constitute common and abundant taxa, particularly in the bacterial phyla Proteobacteria, Actinobacteria, Planctomycetes, Firmicutes, Bacteroidetes, Chloroflexi, and Acidobacteria. The vast majority of OTUs, however, were rare and unique to either soil or rhizosphere communities and differed among locations dozens of kilometers apart. Several soil properties, particularly soil pH and carbon content, were significantly correlated with community diversity measurements. Our results highlight the importance of culture-independent approaches in surveying microbial communities of extreme environments. PMID:22885757

Audit of the global carbon budget: estimate errors and their impact on uptake uncertainty

NASA Astrophysics Data System (ADS)

Ballantyne, A. P.; Andres, R.; Houghton, R.; Stocker, B. D.; Wanninkhof, R.; Anderegg, W.; Cooper, L. A.; DeGrandpre, M.; Tans, P. P.; Miller, J. B.; Alden, C.; White, J. W. C.

2015-04-01

Over the last 5 decades monitoring systems have been developed to detect changes in the accumulation of carbon (C) in the atmosphere and ocean; however, our ability to detect changes in the behavior of the global C cycle is still hindered by measurement and estimate errors. Here we present a rigorous and flexible framework for assessing the temporal and spatial components of estimate errors and their impact on uncertainty in net C uptake by the biosphere. We present a novel approach for incorporating temporally correlated random error into the error structure of emission estimates. Based on this approach, we conclude that the 2σ uncertainties of the atmospheric growth rate have decreased from 1.2 Pg C yr-1 in the 1960s to 0.3 Pg C yr-1 in the 2000s due to an expansion of the atmospheric observation network. The 2σ uncertainties in fossil fuel emissions have increased from 0.3 Pg C yr-1 in the 1960s to almost 1.0 Pg C yr-1 during the 2000s due to differences in national reporting errors and differences in energy inventories. Lastly, while land use emissions have remained fairly constant, their errors still remain high and thus their global C uptake uncertainty is not trivial. Currently, the absolute errors in fossil fuel emissions rival the total emissions from land use, highlighting the extent to which fossil fuels dominate the global C budget. Because errors in the atmospheric growth rate have decreased faster than errors in total emissions have increased, a ~20% reduction in the overall uncertainty of net C global uptake has occurred. Given all the major sources of error in the global C budget that we could identify, we are 93% confident that terrestrial C uptake has increased and 97% confident that ocean C uptake has increased over the last 5 decades. Thus, it is clear that arguably one of the most vital ecosystem services currently provided by the biosphere is the continued removal of approximately half of atmospheric CO2 emissions from the atmosphere, although there are certain environmental costs associated with this service, such as the acidification of ocean waters.

The Influence of Radiosonde 'Age' on TRMM Field Campaign Soundings Humidity Correction

NASA Technical Reports Server (NTRS)

Roy, Biswadev; Halverson, Jeffrey B.; Wang, Jun-Hong

2002-01-01

Hundreds of Vaisala sondes with a RS80-H Humicap thin-film capacitor humidity sensor were launched during the Tropical Rainfall Measuring Mission (TRMM) field campaigns in Large Scale Biosphere-Atmosphere held in Brazil (LBA) and in Kwajalein experiment (KWAJEX) held in the Republic of Marshall Islands. Using Six humidity error correction algorithms by Wang et al., these sondes were corrected for significant dry bias in the RS80-H data. It is further shown that sonde surface temperature error must be corrected for a better representation of the relative humidity. This error becomes prominent due to sensor arm-heating in the first 50-s data.

Rapid and accurate pyrosequencing of angiosperm plastid genomes

PubMed Central

Moore, Michael J; Dhingra, Amit; Soltis, Pamela S; Shaw, Regina; Farmerie, William G; Folta, Kevin M; Soltis, Douglas E

2006-01-01

Background Plastid genome sequence information is vital to several disciplines in plant biology, including phylogenetics and molecular biology. The past five years have witnessed a dramatic increase in the number of completely sequenced plastid genomes, fuelled largely by advances in conventional Sanger sequencing technology. Here we report a further significant reduction in time and cost for plastid genome sequencing through the successful use of a newly available pyrosequencing platform, the Genome Sequencer 20 (GS 20) System (454 Life Sciences Corporation), to rapidly and accurately sequence the whole plastid genomes of the basal eudicot angiosperms Nandina domestica (Berberidaceae) and Platanus occidentalis (Platanaceae). Results More than 99.75% of each plastid genome was simultaneously obtained during two GS 20 sequence runs, to an average depth of coverage of 24.6× in Nandina and 17.3× in Platanus. The Nandina and Platanus plastid genomes shared essentially identical gene complements and possessed the typical angiosperm plastid structure and gene arrangement. To assess the accuracy of the GS 20 sequence, over 45 kilobases of sequence were generated for each genome using conventional sequencing. Overall error rates of 0.043% and 0.031% were observed in GS 20 sequence for Nandina and Platanus, respectively. More than 97% of all observed errors were associated with homopolymer runs, with ~60% of all errors associated with homopolymer runs of 5 or more nucleotides and ~50% of all errors associated with regions of extensive homopolymer runs. No substitution errors were present in either genome. Error rates were generally higher in the single-copy and noncoding regions of both plastid genomes relative to the inverted repeat and coding regions. Conclusion Highly accurate and essentially complete sequence information was obtained for the Nandina and Platanus plastid genomes using the GS 20 System. More importantly, the high accuracy observed in the GS 20 plastid genome sequence was generated for a significant reduction in time and cost over traditional shotgun-based genome sequencing techniques, although with approximately half the coverage of previously reported GS 20 de novo genome sequence. The GS 20 should be broadly applicable to angiosperm plastid genome sequencing, and therefore promises to expand the scale of plant genetic and phylogenetic research dramatically. PMID:16934154

Genome sequencing of bacteria: sequencing, de novo assembly and rapid analysis using open source tools.

PubMed

Kisand, Veljo; Lettieri, Teresa

2013-04-01

De novo genome sequencing of previously uncharacterized microorganisms has the potential to open up new frontiers in microbial genomics by providing insight into both functional capabilities and biodiversity. Until recently, Roche 454 pyrosequencing was the NGS method of choice for de novo assembly because it generates hundreds of thousands of long reads (<450 bps), which are presumed to aid in the analysis of uncharacterized genomes. The array of tools for processing NGS data are increasingly free and open source and are often adopted for both their high quality and role in promoting academic freedom. The error rate of pyrosequencing the Alcanivorax borkumensis genome was such that thousands of insertions and deletions were artificially introduced into the finished genome. Despite a high coverage (~30 fold), it did not allow the reference genome to be fully mapped. Reads from regions with errors had low quality, low coverage, or were missing. The main defect of the reference mapping was the introduction of artificial indels into contigs through lower than 100% consensus and distracting gene calling due to artificial stop codons. No assembler was able to perform de novo assembly comparable to reference mapping. Automated annotation tools performed similarly on reference mapped and de novo draft genomes, and annotated most CDSs in the de novo assembled draft genomes. Free and open source software (FOSS) tools for assembly and annotation of NGS data are being developed rapidly to provide accurate results with less computational effort. Usability is not high priority and these tools currently do not allow the data to be processed without manual intervention. Despite this, genome assemblers now readily assemble medium short reads into long contigs (>97-98% genome coverage). A notable gap in pyrosequencing technology is the quality of base pair calling and conflicting base pairs between single reads at the same nucleotide position. Regardless, using draft whole genomes that are not finished and remain fragmented into tens of contigs allows one to characterize unknown bacteria with modest effort.

Atmospheric CO2 inversions on the mesoscale using data-driven prior uncertainties: quantification of the European terrestrial CO2 fluxes

NASA Astrophysics Data System (ADS)

Kountouris, Panagiotis; Gerbig, Christoph; Rödenbeck, Christian; Karstens, Ute; Koch, Thomas F.; Heimann, Martin

2018-03-01

Optimized biogenic carbon fluxes for Europe were estimated from high-resolution regional-scale inversions, utilizing atmospheric CO2 measurements at 16 stations for the year 2007. Additional sensitivity tests with different data-driven error structures were performed. As the atmospheric network is rather sparse and consequently contains large spatial gaps, we use a priori biospheric fluxes to further constrain the inversions. The biospheric fluxes were simulated by the Vegetation Photosynthesis and Respiration Model (VPRM) at a resolution of 0.1° and optimized against eddy covariance data. Overall we estimate an a priori uncertainty of 0.54 GtC yr-1 related to the poor spatial representation between the biospheric model and the ecosystem sites. The sink estimated from the atmospheric inversions for the area of Europe (as represented in the model domain) ranges between 0.23 and 0.38 GtC yr-1 (0.39 and 0.71 GtC yr-1 up-scaled to geographical Europe). This is within the range of posterior flux uncertainty estimates of previous studies using ground-based observations.

The interaction of the flux errors and transport errors in modeled atmospheric carbon dioxide concentrations

NASA Astrophysics Data System (ADS)

Feng, S.; Lauvaux, T.; Butler, M. P.; Keller, K.; Davis, K. J.; Jacobson, A. R.; Schuh, A. E.; Basu, S.; Liu, J.; Baker, D.; Crowell, S.; Zhou, Y.; Williams, C. A.

2017-12-01

Regional estimates of biogenic carbon fluxes over North America from top-down atmospheric inversions and terrestrial biogeochemical (or bottom-up) models remain inconsistent at annual and sub-annual time scales. While top-down estimates are impacted by limited atmospheric data, uncertain prior flux estimates and errors in the atmospheric transport models, bottom-up fluxes are affected by uncertain driver data, uncertain model parameters and missing mechanisms across ecosystems. This study quantifies both flux errors and transport errors, and their interaction in the CO2 atmospheric simulation. These errors are assessed by an ensemble approach. The WRF-Chem model is set up with 17 biospheric fluxes from the Multiscale Synthesis and Terrestrial Model Intercomparison Project, CarbonTracker-Near Real Time, and the Simple Biosphere model. The spread of the flux ensemble members represents the flux uncertainty in the modeled CO2 concentrations. For the transport errors, WRF-Chem is run using three physical model configurations with three stochastic perturbations to sample the errors from both the physical parameterizations of the model and the initial conditions. Additionally, the uncertainties from boundary conditions are assessed using four CO2 global inversion models which have assimilated tower and satellite CO2 observations. The error structures are assessed in time and space. The flux ensemble members overall overestimate CO2 concentrations. They also show larger temporal variability than the observations. These results suggest that the flux ensemble is overdispersive. In contrast, the transport ensemble is underdispersive. The averaged spatial distribution of modeled CO2 shows strong positive biogenic signal in the southern US and strong negative signals along the eastern coast of Canada. We hypothesize that the former is caused by the 3-hourly downscaling algorithm from which the nighttime respiration dominates the daytime modeled CO2 signals and that the latter is mainly caused by the large-scale transport associated with the jet stream that carries the negative biogenic CO2 signals to the northeastern coast. We apply comprehensive statistics to eliminate outliers. We generate a set of flux perturbations based on pre-calibrated flux ensemble members and apply them to the simulations.

Characterization of the bacterial biodiversity in Pico cheese (an artisanal Azorean food).

PubMed

Riquelme, Cristina; Câmara, Sandra; Dapkevicius, Maria de Lurdes N Enes; Vinuesa, Pablo; da Silva, Célia Costa Gomes; Malcata, F Xavier; Rego, Oldemiro A

2015-01-02

This work presents the first study on the bacterial communities in Pico cheese, a traditional cheese of the Azores (Portugal), made from raw cow's milk. Pyrosequencing of tagged amplicons of the V3-V4 regions of the 16S rDNA and Operational Taxonomic Unit-based (OTU-based) analysis were applied to obtain an overall idea of the microbiota in Pico cheese and to elucidate possible differences between cheese-makers (A, B and C) and maturation times. Pyrosequencing revealed a high bacterial diversity in Pico cheese. Four phyla (Firmicutes, Proteobacteria, Actinobacteria and Bacteroidetes) and 54 genera were identified. The predominant genus was Lactococcus (77% of the sequences). Sequences belonging to major cheese-borne pathogens were not found. Staphylococcus accounted for 0.5% of the sequences. Significant differences in bacterial community composition were observed between cheese-maker B and the other two units that participated in the study. However, OTU analysis identified a set of taxa (Lactococcus, Streptococcus, Acinetobacter, Enterococcus, Lactobacillus, Staphylococcus, Rothia, Pantoea and unclassified genera belonging to the Enterobacteriaceae family) that would represent the core components of artisanal Pico cheese microbiota. A diverse bacterial community was present at early maturation, with an increase in the number of phylotypes up to 2 weeks, followed by a decrease at the end of ripening. The most remarkable trend in abundance patterns throughout ripening was an increase in the number of sequences belonging to the Lactobacillus genus, with a concomitant decrease in Acinetobacter, and Stenotrophomonas. Microbial rank abundance curves showed that Pico cheese's bacterial communities are characterized by a few dominant taxa and many low-abundance, highly diverse taxa that integrate the so-called "rare biosphere". Copyright © 2014 Elsevier B.V. All rights reserved.

Improved Carbon Flux Observations over Urban Areas Using Carbonyl Sulfide (COS) to Differentiate Contributions from Biosphere

NASA Astrophysics Data System (ADS)

Whelan, M.; LaFranchi, B. W.; Bambha, R.; Michelsen, H. A.; Fischer, M. L.; Graven, H. D.; Baker, I. T.; Guilderson, T.; Campbell, J. E.

2016-12-01

Direct measurement and attribution of carbon exchange over urban areas is challenging because of the heterogeneity of the landscape and errors introduced by flux source partitioning. One important contribution to uncertainty is the influence of the urban biosphere on the regional carbon budget. Atmospheric observations of carbonyl sulfide (COS) are an emerging tool for estimating gross primary productivity: COS is consumed in plant leaves by parallel pathways to CO2 uptake, without the additional complexity of an analogous respiration term. This study makes use of COS measurements to better understand fluctuations in total CO2 concentrations over an urban region due to the balance of photosynthesis and respiration. In situ ground-based observations of trace gas concentrations were made from a tower in Livermore, CA, USA, and interpreted with WRF-STILT back trajectories and gridded data sets (e.g. VULCAN, a new anthropogenic COS inventory), supplemented with biosphere models (SiB, CASA-GFED3). CO2, 14CO2, and CO observations were used to first parse the contribution of fossil fuel emissions to total CO2. Changes in the remainder CO2 was differentiated as the sum of biosphere components with associated uncertainties. This approach could be used to better validate carbon emissions reduction measures and ecosytem-based carbon capture projects on the regional scale.

Audit of the global carbon budget: estimate errors and their impact on uptake uncertainty

DOE PAGES

Ballantyne, A. P.; Andres, R.; Houghton, R.; ...

2015-04-30

Over the last 5 decades monitoring systems have been developed to detect changes in the accumulation of carbon (C) in the atmosphere and ocean; however, our ability to detect changes in the behavior of the global C cycle is still hindered by measurement and estimate errors. Here we present a rigorous and flexible framework for assessing the temporal and spatial components of estimate errors and their impact on uncertainty in net C uptake by the biosphere. We present a novel approach for incorporating temporally correlated random error into the error structure of emission estimates. Based on this approach, we concludemore » that the 2σ uncertainties of the atmospheric growth rate have decreased from 1.2 Pg C yr ₋1 in the 1960s to 0.3 Pg C yr ₋1 in the 2000s due to an expansion of the atmospheric observation network. The 2σ uncertainties in fossil fuel emissions have increased from 0.3 Pg C yr ₋1 in the 1960s to almost 1.0 Pg C yr ₋1 during the 2000s due to differences in national reporting errors and differences in energy inventories. Lastly, while land use emissions have remained fairly constant, their errors still remain high and thus their global C uptake uncertainty is not trivial. Currently, the absolute errors in fossil fuel emissions rival the total emissions from land use, highlighting the extent to which fossil fuels dominate the global C budget. Because errors in the atmospheric growth rate have decreased faster than errors in total emissions have increased, a ~20% reduction in the overall uncertainty of net C global uptake has occurred. Given all the major sources of error in the global C budget that we could identify, we are 93% confident that terrestrial C uptake has increased and 97% confident that ocean C uptake has increased over the last 5 decades. Thus, it is clear that arguably one of the most vital ecosystem services currently provided by the biosphere is the continued removal of approximately half of atmospheric CO 2 emissions from the atmosphere, although there are certain environmental costs associated with this service, such as the acidification of ocean waters.« less

Natural Variation in the Carbon Oxidation State and Oxidative Ratio of a Deciduous Forest

NASA Astrophysics Data System (ADS)

Masiello, C. A.; Calligan, L. J.; Gallagher, M. E.; Hockaday, W. C.; Robertson, G. P.

2007-12-01

Here we report natural variability in the oxidative ratio (OR) and carbon oxidation state (Cox) of a temperate, deciduous forest measured on an annual basis via elemental analysis of leaf litter. The OR of the terrestrial biosphere is a key component in O2 -based calculations of the biosphere's uptake of fossil fuel CO2 (eg [ Keeling, et al., 1996]). Ecosystem OR has been assumed to be invariant; however, small OR variations may cause significant shifts in the calculated size of the terrestrial biospheric C sink [ Randerson, et al., 2006]. Accurate measurements of OR are necessary for the accurate apportionment of fossil fuel CO2 between the atmosphere, oceans, and terrestrial biosphere. Ecosystem OR is linearly related to Cox, a parameter which can be easily measured via elemental analysis, calorimetry, or solid state nuclear magnetic resonance [ Masiello, et al., 2007]. We are measuring Cox and OR at the three deciduous forest sites within the Kellogg Biological Station NSF LTER (lter.kbs.msu.edu). We report OR from litter collected from three forest sites from 1998-2003, a time series which covers periods of both normal and low precipitation. We also report error introduced in the Cox to OR conversion via a range of plausible assumptions about ecosystem N cycling. Keeling, R. F., et al. (1996), Global and hemispheric CO2 sinks deduced from changes in atmospheric O2 concentration, Nature, 381, 218-221. Masiello, C.A. et al. (in review 2007) Two new approaches for measuring ecosystem carbon oxidation state and oxidative ratio. J.G.R. Biogeosciences. Randerson, J. T., et al. (2006), Is carbon within the global terrestrial biosphere becoming more oxidized? Implications for trends in atmospheric O2, Global Change Biology, 12, 260-271.

Deep sequencing of hepatitis C virus hypervariable region 1 reveals no correlation between genetic heterogeneity and antiviral treatment outcome

PubMed Central

2014-01-01

Background Hypervariable region 1 (HVR1) contained within envelope protein 2 (E2) gene is the most variable part of HCV genome and its translation product is a major target for the host immune response. Variability within HVR1 may facilitate evasion of the immune response and could affect treatment outcome. The aim of the study was to analyze the impact of HVR1 heterogeneity employing sensitive ultra-deep sequencing, on the outcome of PEG-IFN-α (pegylated interferon α) and ribavirin treatment. Methods HVR1 sequences were amplified from pretreatment serum samples of 25 patients infected with genotype 1b HCV (12 responders and 13 non-responders) and were subjected to pyrosequencing (GS Junior, 454/Roche). Reads were corrected for sequencing error using ShoRAH software, while population reconstruction was done using three different minimal variant frequency cut-offs of 1%, 2% and 5%. Statistical analysis was done using Mann–Whitney and Fisher’s exact tests. Results Complexity, Shannon entropy, nucleotide diversity per site, genetic distance and the number of genetic substitutions were not significantly different between responders and non-responders, when analyzing viral populations at any of the three frequencies (≥1%, ≥2% and ≥5%). When clonal sample was used to determine pyrosequencing error, 4% of reads were found to be incorrect and the most abundant variant was present at a frequency of 1.48%. Use of ShoRAH reduced the sequencing error to 1%, with the most abundant erroneous variant present at frequency of 0.5%. Conclusions While deep sequencing revealed complex genetic heterogeneity of HVR1 in chronic hepatitis C patients, there was no correlation between treatment outcome and any of the analyzed quasispecies parameters. PMID:25016390

Snow surface microbiome on the High Antarctic Plateau (DOME C).

PubMed

Michaud, Luigi; Lo Giudice, Angelina; Mysara, Mohamed; Monsieurs, Pieter; Raffa, Carmela; Leys, Natalie; Amalfitano, Stefano; Van Houdt, Rob

2014-01-01

The cryosphere is an integral part of the global climate system and one of the major habitable ecosystems of Earth's biosphere. These permanently frozen environments harbor diverse, viable and metabolically active microbial populations that represent almost all the major phylogenetic groups. In this study, we investigated the microbial diversity in the surface snow surrounding the Concordia Research Station on the High Antarctic Plateau through a polyphasic approach, including direct prokaryotic quantification by flow cytometry and catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH), and phylogenetic identification by 16S RNA gene clone library sequencing and 454 16S amplicon pyrosequencing. Although the microbial abundance was low (<10(3) cells/ml of snowmelt), concordant results were obtained with the different techniques. The microbial community was mainly composed of members of the Alpha-proteobacteria class (e.g. Kiloniellaceae and Rhodobacteraceae), which is one of the most well-represented bacterial groups in marine habitats, Bacteroidetes (e.g. Cryomorphaceae and Flavobacteriaceae) and Cyanobacteria. Based on our results, polar microorganisms could not only be considered as deposited airborne particles, but as an active component of the snowpack ecology of the High Antarctic Plateau.

Novel insights into methane cycling, lateral gene transfer, and the rare biosphere within carbonate chimneys of the Lost City Hydrothermal Field (Invited)

NASA Astrophysics Data System (ADS)

Brazelton, W. J.; Ludwig, K. A.; Schrenk, M. O.; Kelley, D. S.; Sogin, M. L.; Baross, J. A.

2010-12-01

The Lost City Hydrothermal Field, an ultramafic-hosted system located 15 km west of the Mid-Atlantic Ridge, has experienced at least 30,000 years of hydrothermal activity. Previous studies have shown that its carbonate chimneys form by mixing of ~90°C, pH 9-11 hydrothermal fluids and cold seawater. Flow of methane and hydrogen-rich hydrothermal fluids through the carbonate chimneys supports dense microbial biofilm communities. This presentation will describe recent studies using new techniques that have provided greater insight into the microbial ecology and biogeochemistry of Lost City chimneys. We have investigated the archaeal and bacterial communities of Lost City carbonate chimneys that vary in age between ~30 and ~1200 years, as determined by U-Th isotope systematics. Using next-generation pyrosequencing technology, we collected >200,000 sequences of the V6 region of 16S rRNA genes. This extremely deep sequencing effort enabled detection of very rare organisms as well as abundant organisms detected by previous studies. The taxonomic composition of the archaeal and bacterial communities clearly differed in chimneys of different ages, and many of the rare sequences in young chimneys were more abundant in older chimneys, indicating that members of the rare biosphere can become dominant members of the ecosystem when environmental conditions change. These results suggest that a long history of selection over many cycles of chimney growth has resulted in numerous closely related species at Lost City, each of which is pre-adapted to a particular set of re-occurring environmental conditions. In this model, the rare biosphere can be considered a repository for genes that are not currently advantageous but have been in the past and may be again in the future. Interestingly, metagenomic sequencing at Lost City has indicated the potential for frequent lateral gene transfer among organisms inhabiting the chimney biofilms. Specifically, the Lost City metagenomic dataset contains the highest proportion of genes encoding transposases (the enzymes required for lateral gene transfer) of any metagenome yet reported. Therefore, we intend to test the hypothesis that the rare biosphere can serve as a source of genes that can be laterally transferred to and benefit organisms of ecological importance. The metagenomic dataset also provides clues regarding the physiology of the dominant organisms. For example, bacterial genes associated with sulfide and sulfur oxidation are present. We have also detected archaeal genes associated with nitrogen fixation and conversion of acetate to methane. Laboratory incubation experiments have shown that anaerobic methane oxidation also occurs in Lost City chimney biofilms. We will present a conceptual model of biogeochemical reactions expected to occur in Lost City biofilms.

Biospheric energization and stability

NASA Astrophysics Data System (ADS)

Budding, E.; Ozel, M. E.; Gunduz, G.

2013-09-01

We utilize the physical properties of a hypothetical molecular schema giving rise to an autocatalytic biosphere. A key concept is the driving of terrestrial life as a parametric oscillation: i.e. that the biosphere behaves fundamentally as an oscillatory system into which solar energy is diurnally deposited. The schema, containing 'A, B and C' type components acting together in a 'bottom-up' driving mechanism, underlies all biospheric superstructure. Surviving modes of the oscillation are consistent with Darwinian organization, or hierarchical structures appearing to have top-down propagation through the growth of cellular replication. The model was detailed by Budding et al (2012), where experimental support from the work of Powner et al (2009) is presented, as well as suggestions on supportive fossil evidence. Although the growth in total energization is very slow in this model, it is important to notice its exponential character, suggestive of potential instability. The model is applicable to generally expectable processes on planets, including zonal segregation, complexity growth and Haeckel's biogenic principle within surviving life-forms. Fermi's exobiological paradox can be resolved in terms of the exponential growth and low L solutions of Drake's equation. Feasible values for the particular growth of selected species (the human one in herelevent terrestrial case) allow for L to be less than a few  100 y, recalling Rees' (2004) 'final century' discussion. This arises when the species' disposable energization attains a value comparable to that of the total available daily driving energy. At that point, accidental, or stochastic disturbances of this species' energy ("error") can significantly disrupt the daily driving mechanism.

Regional climate modeling over the Maritime Continent: Assessment of RegCM3-BATS1e and RegCM3-IBIS

NASA Astrophysics Data System (ADS)

Gianotti, R. L.; Zhang, D.; Eltahir, E. A.

2010-12-01

Despite its importance to global rainfall and circulation processes, the Maritime Continent remains a region that is poorly simulated by climate models. Relatively few studies have been undertaken using a model with fine enough resolution to capture the small-scale spatial heterogeneity of this region and associated land-atmosphere interactions. These studies have shown that even regional climate models (RCMs) struggle to reproduce the climate of this region, particularly the diurnal cycle of rainfall. This study builds on previous work by undertaking a more thorough evaluation of RCM performance in simulating the timing and intensity of rainfall over the Maritime Continent, with identification of major sources of error. An assessment was conducted of the Regional Climate Model Version 3 (RegCM3) used in a coupled system with two land surface schemes: Biosphere Atmosphere Transfer System Version 1e (BATS1e) and Integrated Biosphere Simulator (IBIS). The model’s performance in simulating precipitation was evaluated against the 3-hourly TRMM 3B42 product, with some validation provided of this TRMM product against ground station meteorological data. It is found that the model suffers from three major errors in the rainfall histogram: underestimation of the frequency of dry periods, overestimation of the frequency of low intensity rainfall, and underestimation of the frequency of high intensity rainfall. Additionally, the model shows error in the timing of the diurnal rainfall peak, particularly over land surfaces. These four errors were largely insensitive to the choice of boundary conditions, convective parameterization scheme or land surface scheme. The presence of a wet or dry bias in the simulated volumes of rainfall was, however, dependent on the choice of convection scheme and boundary conditions. This study also showed that the coupled model system has significant error in overestimation of latent heat flux and evapotranspiration from the land surface, and specifically overestimation of interception loss with concurrent underestimation of transpiration, irrespective of the land surface scheme used. Discussion of the origin of these errors is provided, with some suggestions for improvement.

Multiplex pyrosequencing of InDel markers for forensic DNA analysis.

PubMed

Bus, Magdalena M; Karas, Ognjen; Allen, Marie

2016-12-01

The capillary electrophoresis (CE) technology is commonly used for fragment length separation of markers in forensic DNA analysis. In this study, pyrosequencing technology was used as an alternative and rapid tool for the analysis of biallelic InDel (insertion/deletion) markers for individual identification. The DNA typing is based on a subset of the InDel markers that are included in the Investigator ® DIPplex Kit, which are sequenced in a multiplex pyrosequencing analysis. To facilitate the analysis of degraded DNA, the polymerase chain reaction (PCR) fragments were kept short in the primer design. Samples from individuals of Swedish origin were genotyped using the pyrosequencing strategy and analysis of the Investigator ® DIPplex markers with CE. A comparison between the pyrosequencing and CE data revealed concordant results demonstrating a robust and correct genotyping by pyrosequencing. Using optimal marker combination and a directed dispensation strategy, five markers could be multiplexed and analyzed simultaneously. In this proof-of-principle study, we demonstrate that multiplex InDel pyrosequencing analysis is possible. However, further studies on degraded samples, lower DNA quantities, and mixtures will be required to fully optimize InDel analysis by pyrosequencing for forensic applications. Overall, although CE analysis is implemented in most forensic laboratories, multiplex InDel pyrosequencing offers a cost-effective alternative for some applications. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

[Sensitivity and specificity of nested PCR pyrosequencing in hepatitis B virus drug resistance gene testing].

PubMed

Sun, Shumei; Zhou, Hao; Zhou, Bin; Hu, Ziyou; Hou, Jinlin; Sun, Jian

2012-05-01

To evaluate the sensitivity and specificity of nested PCR combined with pyrosequencing in the detection of HBV drug-resistance gene. RtM204I (ATT) mutant and rtM204 (ATG) nonmutant plasmids mixed at different ratios were detected for mutations using nested-PCR combined with pyrosequencing, and the results were compared with those by conventional PCR pyrosequencing to analyze the linearity and consistency of the two methods. Clinical specimens with different viral loads were examined for drug-resistant mutations using nested PCR pyrosequencing and nested PCR combined with dideoxy sequencing (Sanger) for comparison of the detection sensitivity and specificity. The fitting curves demonstrated good linearity of both conventional PCR pyrosequencing and nested PCR pyrosequencing (R(2)>0.99, P<0.05). Nested PCR showed a better consistency with the predicted value than conventional PCR, and was superior to conventional PCR for detection of samples containing 90% mutant plasmid. In the detection of clinical specimens, Sanger sequencing had a significantly lower sensitivity than nested PCR pyrosequencing (92% vs 100%, P<0.01). The detection sensitivity of Sanger sequencing varied with the viral loads, especially in samples with low viral copies (HBV DNA ≤3log10 copies/ml), where the sensitivity was 78%, significantly lower than that of pyrosequencing (100%, P<0.01). Neither of the two methods yielded positive results for the negative control samples, suggesting their good specificity. Compared with nested PCR and Sanger sequencing method, nested PCR pyrosequencing has a higher sensitivity especially in clinical specimens with low viral copies, which can be important for early detection of HBV mutant strains and hence more effective clinical management.

Assessment of Provisional MODIS-derived Surfaces Related to the Global Carbon Cycle

NASA Astrophysics Data System (ADS)

Cohen, W. B.; Maiersperger, T. K.; Turner, D. P.; Gower, S. T.; Kennedy, R. E.; Running, S. W.

2002-12-01

The global carbon cycle is one of the most important foci of an emerging global biosphere monitoring system. A key component of such a system is the MODIS sensor, onboard the Terra satellite platform. Biosphere monitoring requires an integrated program of satellite observations, Earth-system models, and in situ data. Related to the carbon cycle, MODIS science teams routinely develop a variety of global surfaces such as land cover, leaf area index, and net primary production using MODIS data and functional algorithms. The quality of these surfaces must be evaluated to determine their effectiveness for global biosphere monitoring. A project called BigFoot (http://www.fsl.orst.edu/larse/bigfoot/) is an organized effort across nine biomes to assess the quality of the abovementioned surfaces: (1) Arctic tundra; (2) boreal evergreen needle-leaved forest; temperate (3) cropland, (4) grassland, (5) evergreen needle-leaved forest, and (6) deciduous broad-leaved forest; desert (7) grassland and (8) shrubland; and (9) tropical evergreen broad-leaved forest. Each biome is represented by a site that has an eddy-covariance flux tower that measures water vapor and CO2 fluxes. Flux tower footprints are relatively small-approximately 1 km2. BigFoot characterizes 25 km2 around each tower, using field data, Landsat ETM+ image data, and ecosystem process models. Our innovative field sampling design incorporates a nested spatial series to facilitate geostatistical analyses, samples the ecological variability at a site, and is logistically efficient. Field data are used both to develop site-specific algorithms for mapping/modeling the variables of interest and to characterize the errors in derived BigFoot surfaces. Direct comparisons of BigFoot- and MODIS-derived surfaces are made to help understand the sources of error in MODIS-derived surfaces and to facilitate improvements to MODIS algorithms. Results from four BigFoot sites will be presented.

Denoising DNA deep sequencing data—high-throughput sequencing errors and their correction

PubMed Central

Laehnemann, David; Borkhardt, Arndt

2016-01-01

Characterizing the errors generated by common high-throughput sequencing platforms and telling true genetic variation from technical artefacts are two interdependent steps, essential to many analyses such as single nucleotide variant calling, haplotype inference, sequence assembly and evolutionary studies. Both random and systematic errors can show a specific occurrence profile for each of the six prominent sequencing platforms surveyed here: 454 pyrosequencing, Complete Genomics DNA nanoball sequencing, Illumina sequencing by synthesis, Ion Torrent semiconductor sequencing, Pacific Biosciences single-molecule real-time sequencing and Oxford Nanopore sequencing. There is a large variety of programs available for error removal in sequencing read data, which differ in the error models and statistical techniques they use, the features of the data they analyse, the parameters they determine from them and the data structures and algorithms they use. We highlight the assumptions they make and for which data types these hold, providing guidance which tools to consider for benchmarking with regard to the data properties. While no benchmarking results are included here, such specific benchmarks would greatly inform tool choices and future software development. The development of stand-alone error correctors, as well as single nucleotide variant and haplotype callers, could also benefit from using more of the knowledge about error profiles and from (re)combining ideas from the existing approaches presented here. PMID:26026159

The biosphere rules.

PubMed

Unruh, Gregory C

2008-02-01

Sustainability, defined by natural scientists as the capacity of healthy ecosystems to function indefinitely, has become a clarion call for business. Leading companies have taken high-profile steps toward achieving it: Wal-Mart, for example, with its efforts to reduce packaging waste, and Nike, which has removed toxic chemicals from its shoes. But, says Unruh, the director of Thunderbird's Lincoln Center for Ethics in Global Management, sustainability is more than an endless journey of incremental steps. It is a destination, for which the biosphere of planet Earth--refined through billions of years of trial and error--is a perfect model. Unruh distills some lessons from the biosphere into three rules: Use a parsimonious palette. Managers can rethink their sourcing strategies and dramatically simplify the number and types of materials their companies use in production, making recycling cost-effective. After the furniture manufacturer Herman Miller discovered that its leading desk chair had 200 components made from more than 800 chemical compounds, it designed an award-winning successor whose far more limited materials palette is 96% recyclable. Cycle up, virtuously. Manufacturers should design recovery value into their products at the outset. Shaw Industries, for example, recycles the nylon fiber from its worn-out carpet into brand-new carpet tile. Exploit the power of platforms. Platform design in industry tends to occur at the component level--but the materials in those components constitute a more fundamental platform. Patagonia, by recycling Capilene brand performance underwear, has achieved energy costs 76% below those for virgin sourcing. Biosphere rules can teach companies how to build ecologically friendly products that both reduce manufacturing costs and prove highly attractive to consumers. And managers need not wait for a green technological revolution to implement them.

Snow Surface Microbiome on the High Antarctic Plateau (DOME C)

PubMed Central

Michaud, Luigi; Lo Giudice, Angelina; Mysara, Mohamed; Monsieurs, Pieter; Raffa, Carmela; Leys, Natalie; Amalfitano, Stefano; Van Houdt, Rob

2014-01-01

The cryosphere is an integral part of the global climate system and one of the major habitable ecosystems of Earth's biosphere. These permanently frozen environments harbor diverse, viable and metabolically active microbial populations that represent almost all the major phylogenetic groups. In this study, we investigated the microbial diversity in the surface snow surrounding the Concordia Research Station on the High Antarctic Plateau through a polyphasic approach, including direct prokaryotic quantification by flow cytometry and catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH), and phylogenetic identification by 16S RNA gene clone library sequencing and 454 16S amplicon pyrosequencing. Although the microbial abundance was low (<103 cells/ml of snowmelt), concordant results were obtained with the different techniques. The microbial community was mainly composed of members of the Alpha-proteobacteria class (e.g. Kiloniellaceae and Rhodobacteraceae), which is one of the most well-represented bacterial groups in marine habitats, Bacteroidetes (e.g. Cryomorphaceae and Flavobacteriaceae) and Cyanobacteria. Based on our results, polar microorganisms could not only be considered as deposited airborne particles, but as an active component of the snowpack ecology of the High Antarctic Plateau. PMID:25101779

Efficient error correction for next-generation sequencing of viral amplicons

PubMed Central

2012-01-01

Background Next-generation sequencing allows the analysis of an unprecedented number of viral sequence variants from infected patients, presenting a novel opportunity for understanding virus evolution, drug resistance and immune escape. However, sequencing in bulk is error prone. Thus, the generated data require error identification and correction. Most error-correction methods to date are not optimized for amplicon analysis and assume that the error rate is randomly distributed. Recent quality assessment of amplicon sequences obtained using 454-sequencing showed that the error rate is strongly linked to the presence and size of homopolymers, position in the sequence and length of the amplicon. All these parameters are strongly sequence specific and should be incorporated into the calibration of error-correction algorithms designed for amplicon sequencing. Results In this paper, we present two new efficient error correction algorithms optimized for viral amplicons: (i) k-mer-based error correction (KEC) and (ii) empirical frequency threshold (ET). Both were compared to a previously published clustering algorithm (SHORAH), in order to evaluate their relative performance on 24 experimental datasets obtained by 454-sequencing of amplicons with known sequences. All three algorithms show similar accuracy in finding true haplotypes. However, KEC and ET were significantly more efficient than SHORAH in removing false haplotypes and estimating the frequency of true ones. Conclusions Both algorithms, KEC and ET, are highly suitable for rapid recovery of error-free haplotypes obtained by 454-sequencing of amplicons from heterogeneous viruses. The implementations of the algorithms and data sets used for their testing are available at: http://alan.cs.gsu.edu/NGS/?q=content/pyrosequencing-error-correction-algorithm PMID:22759430

Efficient error correction for next-generation sequencing of viral amplicons.

PubMed

Skums, Pavel; Dimitrova, Zoya; Campo, David S; Vaughan, Gilberto; Rossi, Livia; Forbi, Joseph C; Yokosawa, Jonny; Zelikovsky, Alex; Khudyakov, Yury

2012-06-25

Next-generation sequencing allows the analysis of an unprecedented number of viral sequence variants from infected patients, presenting a novel opportunity for understanding virus evolution, drug resistance and immune escape. However, sequencing in bulk is error prone. Thus, the generated data require error identification and correction. Most error-correction methods to date are not optimized for amplicon analysis and assume that the error rate is randomly distributed. Recent quality assessment of amplicon sequences obtained using 454-sequencing showed that the error rate is strongly linked to the presence and size of homopolymers, position in the sequence and length of the amplicon. All these parameters are strongly sequence specific and should be incorporated into the calibration of error-correction algorithms designed for amplicon sequencing. In this paper, we present two new efficient error correction algorithms optimized for viral amplicons: (i) k-mer-based error correction (KEC) and (ii) empirical frequency threshold (ET). Both were compared to a previously published clustering algorithm (SHORAH), in order to evaluate their relative performance on 24 experimental datasets obtained by 454-sequencing of amplicons with known sequences. All three algorithms show similar accuracy in finding true haplotypes. However, KEC and ET were significantly more efficient than SHORAH in removing false haplotypes and estimating the frequency of true ones. Both algorithms, KEC and ET, are highly suitable for rapid recovery of error-free haplotypes obtained by 454-sequencing of amplicons from heterogeneous viruses.The implementations of the algorithms and data sets used for their testing are available at: http://alan.cs.gsu.edu/NGS/?q=content/pyrosequencing-error-correction-algorithm.

A Regional CO2 Observing System Simulation Experiment Using ASCENDS Observations and WRF-STILT Footprints

NASA Technical Reports Server (NTRS)

Wang, James S.; Kawa, S. Randolph; Eluszkiewicz, Janusz; Collatz, G. J.; Mountain, Marikate; Henderson, John; Nehrkorn, Thomas; Aschbrenner, Ryan; Zaccheo, T. Scott

2012-01-01

Knowledge of the spatiotemporal variations in emissions and uptake of CO2 is hampered by sparse measurements. The recent advent of satellite measurements of CO2 concentrations is increasing the density of measurements, and the future mission ASCENDS (Active Sensing of CO2 Emissions over Nights, Days and Seasons) will provide even greater coverage and precision. Lagrangian atmospheric transport models run backward in time can quantify surface influences ("footprints") of diverse measurement platforms and are particularly well suited for inverse estimation of regional surface CO2 fluxes at high resolution based on satellite observations. We utilize the STILT Lagrangian particle dispersion model, driven by WRF meteorological fields at 40-km resolution, in a Bayesian synthesis inversion approach to quantify the ability of ASCENDS column CO2 observations to constrain fluxes at high resolution. This study focuses on land-based biospheric fluxes, whose uncertainties are especially large, in a domain encompassing North America. We present results based on realistic input fields for 2007. Pseudo-observation random errors are estimated from backscatter and optical depth measured by the CALIPSO satellite. We estimate a priori flux uncertainties based on output from the CASA-GFED (v.3) biosphere model and make simple assumptions about spatial and temporal error correlations. WRF-STILT footprints are convolved with candidate vertical weighting functions for ASCENDS. We find that at a horizontal flux resolution of 1 degree x 1 degree, ASCENDS observations are potentially able to reduce average weekly flux uncertainties by 0-8% in July, and 0-0.5% in January (assuming an error of 0.5 ppm at the Railroad Valley reference site). Aggregated to coarser resolutions, e.g. 5 degrees x 5 degrees, the uncertainty reductions are larger and more similar to those estimated in previous satellite data observing system simulation experiments.

Pyrosequencing-based quantitative measurement of CALR mutation allele burdens and their clinical implications in patients with myeloproliferative neoplasms.

PubMed

Oh, Yejin; Song, Ik-Chan; Kim, Jimyung; Kwon, Gye Cheol; Koo, Sun Hoe; Kim, Seon Young

2018-05-01

We developed a pyrosequencing-based method for the quantification of CALR mutations and compared the results using Sanger sequencing, fragment length analysis (FLA), digital-droplet PCR (ddPCR), and next-generation sequencing (NGS). Method validation studies were performed using cloned plasmid controls. Samples from 24 patients with myeloproliferative neoplasms were evaluated. Among the 24 patients, 15 had CALR mutations (7 type 1, 2 type 2, and 6 other mutations). The type 1 or type 2 mutation-positive results from pyrosequencing exhibited 100% concordance with the Sanger sequencing results. One novel CALR mutation was not detected by pyrosequencing. The CALR mutation allele burdens measured by pyrosequencing were slightly lower than those measured by FLA but slightly higher than the results obtained using ddPCR. Pyrosequencing exhibited high correlations with both methods. The mutation allele burdens estimated by NGS were significantly lower than those measured by pyrosequencing. An increased CALR mutation allele burden was associated with overt primary myelofibrosis. Patients with >70% mutation allele burdens in myeloid cells had a significantly longer time from diagnosis (P = 0.007), more bone marrow fibrosis (P = 0.010), and lower hemoglobin (P = 0.007). Pyrosequencing was a useful rapid sequencing method to determine the burden of CALR mutations. Copyright © 2018 Elsevier B.V. All rights reserved.

Rapid Molecular Identification of Pathogenic Yeasts by Pyrosequencing Analysis of 35 Nucleotides of Internal Transcribed Spacer 2 ▿

PubMed Central

Borman, Andrew M.; Linton, Christopher J.; Oliver, Debra; Palmer, Michael D.; Szekely, Adrien; Johnson, Elizabeth M.

2010-01-01

Rapid identification of yeast species isolates from clinical samples is particularly important given their innately variable antifungal susceptibility profiles. Here, we have evaluated the utility of pyrosequencing analysis of a portion of the internal transcribed spacer 2 region (ITS2) for identification of pathogenic yeasts. A total of 477 clinical isolates encompassing 43 different fungal species were subjected to pyrosequencing analysis in a strictly blinded study. The molecular identifications produced by pyrosequencing were compared with those obtained using conventional biochemical tests (AUXACOLOR2) and following PCR amplification and sequencing of the D1-D2 portion of the nuclear 28S large rRNA gene. More than 98% (469/477) of isolates encompassing 40 of the 43 fungal species tested were correctly identified by pyrosequencing of only 35 bp of ITS2. Moreover, BLAST searches of the public synchronized databases with the ITS2 pyrosequencing signature sequences revealed that there was only minimal sequence redundancy in the ITS2 under analysis. In all cases, the pyrosequencing signature sequences were unique to the yeast species (or species complex) under investigation. Finally, when pyrosequencing was combined with the Whatman FTA paper technology for the rapid extraction of fungal genomic DNA, molecular identification could be accomplished within 6 h from the time of starting from pure cultures. PMID:20702674

Geosphere-Biosphere Interactions in Bio-Activity Volcanic Lakes: Evidences from Hule and Rìo Cuarto (Costa Rica)

PubMed Central

Cabassi, Jacopo; Tassi, Franco; Mapelli, Francesca; Borin, Sara; Calabrese, Sergio; Rouwet, Dmitri; Chiodini, Giovanni; Marasco, Ramona; Chouaia, Bessem; Avino, Rosario; Vaselli, Orlando; Pecoraino, Giovannella; Capecchiacci, Francesco; Bicocchi, Gabriele; Caliro, Stefano; Ramirez, Carlos; Mora-Amador, Raul

2014-01-01

Hule and Río Cuarto are maar lakes located 11 and 18 km N of Poás volcano along a 27 km long fracture zone, in the Central Volcanic Range of Costa Rica. Both lakes are characterized by a stable thermic and chemical stratification and recently they were affected by fish killing events likely related to the uprising of deep anoxic waters to the surface caused by rollover phenomena. The vertical profiles of temperature, pH, redox potential, chemical and isotopic compositions of water and dissolved gases, as well as prokaryotic diversity estimated by DNA fingerprinting and massive 16S rRNA pyrosequencing along the water column of the two lakes, have highlighted that different bio-geochemical processes occur in these meromictic lakes. Although the two lakes host different bacterial and archaeal phylogenetic groups, water and gas chemistry in both lakes is controlled by the same prokaryotic functions, especially regarding the CO2-CH4 cycle. Addition of hydrothermal CO2 through the bottom of the lakes plays a fundamental priming role in developing a stable water stratification and fuelling anoxic bacterial and archaeal populations. Methanogens and methane oxidizers as well as autotrophic and heterotrophic aerobic bacteria responsible of organic carbon recycling resulted to be stratified with depth and strictly related to the chemical-physical conditions and availability of free oxygen, affecting both the CO2 and CH4 chemical concentrations and their isotopic compositions along the water column. Hule and Río Cuarto lakes were demonstrated to contain a CO2 (CH4, N2)-rich gas reservoir mainly controlled by the interactions occurring between geosphere and biosphere. Thus, we introduced the term of bio-activity volcanic lakes to distinguish these lakes, which have analogues worldwide (e.g. Kivu: D.R.C.-Rwanda; Albano, Monticchio and Averno: Italy; Pavin: France) from volcanic lakes only characterized by geogenic CO2 reservoir such as Nyos and Monoun (Cameroon). PMID:25058537

Geosphere-biosphere interactions in bio-activity volcanic lakes: evidences from Hule and Rìo Cuarto (Costa Rica).

PubMed

Cabassi, Jacopo; Tassi, Franco; Mapelli, Francesca; Borin, Sara; Calabrese, Sergio; Rouwet, Dmitri; Chiodini, Giovanni; Marasco, Ramona; Chouaia, Bessem; Avino, Rosario; Vaselli, Orlando; Pecoraino, Giovannella; Capecchiacci, Francesco; Bicocchi, Gabriele; Caliro, Stefano; Ramirez, Carlos; Mora-Amador, Raul

2014-01-01

Hule and Río Cuarto are maar lakes located 11 and 18 km N of Poás volcano along a 27 km long fracture zone, in the Central Volcanic Range of Costa Rica. Both lakes are characterized by a stable thermic and chemical stratification and recently they were affected by fish killing events likely related to the uprising of deep anoxic waters to the surface caused by rollover phenomena. The vertical profiles of temperature, pH, redox potential, chemical and isotopic compositions of water and dissolved gases, as well as prokaryotic diversity estimated by DNA fingerprinting and massive 16S rRNA pyrosequencing along the water column of the two lakes, have highlighted that different bio-geochemical processes occur in these meromictic lakes. Although the two lakes host different bacterial and archaeal phylogenetic groups, water and gas chemistry in both lakes is controlled by the same prokaryotic functions, especially regarding the CO2-CH4 cycle. Addition of hydrothermal CO2 through the bottom of the lakes plays a fundamental priming role in developing a stable water stratification and fuelling anoxic bacterial and archaeal populations. Methanogens and methane oxidizers as well as autotrophic and heterotrophic aerobic bacteria responsible of organic carbon recycling resulted to be stratified with depth and strictly related to the chemical-physical conditions and availability of free oxygen, affecting both the CO2 and CH4 chemical concentrations and their isotopic compositions along the water column. Hule and Río Cuarto lakes were demonstrated to contain a CO2 (CH4, N2)-rich gas reservoir mainly controlled by the interactions occurring between geosphere and biosphere. Thus, we introduced the term of bio-activity volcanic lakes to distinguish these lakes, which have analogues worldwide (e.g. Kivu: D.R.C.-Rwanda; Albano, Monticchio and Averno: Italy; Pavin: France) from volcanic lakes only characterized by geogenic CO2 reservoir such as Nyos and Monoun (Cameroon).

Diversity and homogeneity of oral microbiota in healthy Korean pre-school children using pyrosequencing.

PubMed

Lee, Soo Eon; Nam, Ok Hyung; Lee, Hyo-Seol; Choi, Sung Chul

2016-07-01

Objectives The purpose of this study was designed to identify the oral microbiota in healthy Korean pre-school children using pyrosequencing. Materials and methods Dental plaque samples were obtained form 10 caries-free pre-school children. The samples were analysed using pyrosequencing. Results The pyrosequencing analysis revealed that, at the phylum level, Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria and Fusobacteria showed high abundance. Also, predominant genera were identified as core microbiome, such as Streptococcus, Neisseria, Capnocytophaga, Haemophilus and Veilonella. Conclusions The diversity and homogeneity was shown in the dental plaque microbiota in healthy Korean pre-school children.

NASA's Carbon Monitoring System Flux-Pilot Project: A Multi-Component Analysis System for Carbon-Cycle Research and Monitoring

NASA Technical Reports Server (NTRS)

Pawson, S.; Gunson, M.; Potter, C.; Jucks, K.

2012-01-01

The importance of greenhouse gas increases for climate motivates NASA s observing strategy for CO2 from space, including the forthcoming Orbiting Carbon Observatory (OCO-2) mission. Carbon cycle monitoring, including attribution of atmospheric concentrations to regional emissions and uptake, requires a robust modeling and analysis infrastructure to optimally extract information from the observations. NASA's Carbon-Monitoring System Flux-Pilot Project (FPP) is a prototype for such analysis, combining a set of unique tools to facilitate analysis of atmospheric CO2 along with fluxes between the atmosphere and the terrestrial biosphere or ocean. NASA's analysis system is unique, in that it combines information and expertise from the land, oceanic, and atmospheric branches of the carbon cycle and includes some estimates of uncertainty. Numerous existing space-based missions provide information of relevance to the carbon cycle. This study describes the components of the FPP framework, assessing the realism of computed fluxes, thus providing the basis for research and monitoring applications. Fluxes are computed using data-constrained terrestrial biosphere models and physical ocean models, driven by atmospheric observations and assimilating ocean-color information. Use of two estimates provides a measure of uncertainty in the fluxes. Along with inventories of other emissions, these data-derived fluxes are used in transport models to assess their consistency with atmospheric CO2 observations. Closure is achieved by using a four-dimensional data assimilation (inverse) approach that adjusts the terrestrial biosphere fluxes to make them consistent with the atmospheric CO2 observations. Results will be shown, illustrating the year-to-year variations in land biospheric and oceanic fluxes computed in the FPP. The signals of these surface-flux variations on atmospheric CO2 will be isolated using forward modeling tools, which also incorporate estimates of transport error. The results will be discussed in the context of interannual variability of observed atmospheric CO2 distributions.

SNP-based real-time pyrosequencing as a sensitive and specific tool for identification and differentiation of Rickettsia species in Ixodes ricinus ticks.

PubMed

Janecek, Elisabeth; Streichan, Sabine; Strube, Christina

2012-10-18

Rickettsioses are caused by pathogenic species of the genus Rickettsia and play an important role as emerging diseases. The bacteria are transmitted to mammal hosts including humans by arthropod vectors. Since detection, especially in tick vectors, is usually based on PCR with genus-specific primers to include different occurring Rickettsia species, subsequent species identification is mainly achieved by Sanger sequencing. In the present study a real-time pyrosequencing approach was established with the objective to differentiate between species occurring in German Ixodes ticks, which are R. helvetica, R. monacensis, R. massiliae, and R. felis. Tick material from a quantitative real-time PCR (qPCR) based study on Rickettsia-infections in I. ricinus allowed direct comparison of both sequencing techniques, Sanger and real-time pyrosequencing. A sequence stretch of rickettsial citrate synthase (gltA) gene was identified to contain divergent single nucleotide polymorphism (SNP) sites suitable for Rickettsia species differentiation. Positive control plasmids inserting the respective target sequence of each Rickettsia species of interest were constructed for initial establishment of the real-time pyrosequencing approach using Qiagen's PSQ 96MA Pyrosequencing System operating in a 96-well format. The approach included an initial amplification reaction followed by the actual pyrosequencing, which is traceable by pyrograms in real-time. Afterwards, real-time pyrosequencing was applied to 263 Ixodes tick samples already detected Rickettsia-positive in previous qPCR experiments. Establishment of real-time pyrosequencing using positive control plasmids resulted in accurate detection of all SNPs in all included Rickettsia species. The method was then applied to 263 Rickettsia-positive Ixodes ricinus samples, of which 153 (58.2%) could be identified for their species (151 R. helvetica and 2 R. monacensis) by previous custom Sanger sequencing. Real-time pyrosequencing identified all Sanger-determined ticks as well as 35 previously undifferentiated ticks resulting in a total number of 188 (71.5%) identified samples. Pyrosequencing sensitivity was found to be strongly dependent on gltA copy numbers in the reaction setup. Whereas less than 101 copies in the initial amplification reaction resulted in identification of 15.1% of the samples only, the percentage increased to 54.2% at 101-102 copies, to 95.6% at >102-103 copies and reached 100% samples identified for their Rickettsia species if more than 103 copies were present in the template. The established real-time pyrosequencing approach represents a reliable method for detection and differentiation of Rickettsia spp. present in I. ricinus diagnostic material and prevalence studies. Furthermore, the method proved to be faster, more cost-effective as well as more sensitive than custom Sanger sequencing with simultaneous high specificity.

454 Pyrosequencing to Describe Microbial Eukaryotic Community Composition, Diversity and Relative Abundance: A Test for Marine Haptophytes

PubMed Central

Egge, Elianne; Bittner, Lucie; Andersen, Tom; Audic, Stéphane; de Vargas, Colomban; Edvardsen, Bente

2013-01-01

Next generation sequencing of ribosomal DNA is increasingly used to assess the diversity and structure of microbial communities. Here we test the ability of 454 pyrosequencing to detect the number of species present, and assess the relative abundance in terms of cell numbers and biomass of protists in the phylum Haptophyta. We used a mock community consisting of equal number of cells of 11 haptophyte species and compared targeting DNA and RNA/cDNA, and two different V4 SSU rDNA haptophyte-biased primer pairs. Further, we tested four different bioinformatic filtering methods to reduce errors in the resulting sequence dataset. With sequencing depth of 11000–20000 reads and targeting cDNA with Haptophyta specific primers Hap454 we detected all 11 species. A rarefaction analysis of expected number of species recovered as a function of sampling depth suggested that minimum 1400 reads were required here to recover all species in the mock community. Relative read abundance did not correlate to relative cell numbers. Although the species represented with the largest biomass was also proportionally most abundant among the reads, there was generally a weak correlation between proportional read abundance and proportional biomass of the different species, both with DNA and cDNA as template. The 454 sequencing generated considerable spurious diversity, and more with cDNA than DNA as template. With initial filtering based only on match with barcode and primer we observed 100-fold more operational taxonomic units (OTUs) at 99% similarity than the number of species present in the mock community. Filtering based on quality scores, or denoising with PyroNoise resulted in ten times more OTU99% than the number of species. Denoising with AmpliconNoise reduced the number of OTU99% to match the number of species present in the mock community. Based on our analyses, we propose a strategy to more accurately depict haptophyte diversity using 454 pyrosequencing. PMID:24069303

Pyrosequencing for Microbial Identification and Characterization

PubMed Central

Cummings, Patrick J.; Ahmed, Ray; Durocher, Jeffrey A.; Jessen, Adam; Vardi, Tamar; Obom, Kristina M.

2013-01-01

Pyrosequencing is a versatile technique that facilitates microbial genome sequencing that can be used to identify bacterial species, discriminate bacterial strains and detect genetic mutations that confer resistance to anti-microbial agents. The advantages of pyrosequencing for microbiology applications include rapid and reliable high-throughput screening and accurate identification of microbes and microbial genome mutations. Pyrosequencing involves sequencing of DNA by synthesizing the complementary strand a single base at a time, while determining the specific nucleotide being incorporated during the synthesis reaction. The reaction occurs on immobilized single stranded template DNA where the four deoxyribonucleotides (dNTP) are added sequentially and the unincorporated dNTPs are enzymatically degraded before addition of the next dNTP to the synthesis reaction. Detection of the specific base incorporated into the template is monitored by generation of chemiluminescent signals. The order of dNTPs that produce the chemiluminescent signals determines the DNA sequence of the template. The real-time sequencing capability of pyrosequencing technology enables rapid microbial identification in a single assay. In addition, the pyrosequencing instrument, can analyze the full genetic diversity of anti-microbial drug resistance, including typing of SNPs, point mutations, insertions, and deletions, as well as quantification of multiple gene copies that may occur in some anti-microbial resistance patterns. PMID:23995536

Pyrosequencing for microbial identification and characterization.

PubMed

Cummings, Patrick J; Ahmed, Ray; Durocher, Jeffrey A; Jessen, Adam; Vardi, Tamar; Obom, Kristina M

2013-08-22

Pyrosequencing is a versatile technique that facilitates microbial genome sequencing that can be used to identify bacterial species, discriminate bacterial strains and detect genetic mutations that confer resistance to anti-microbial agents. The advantages of pyrosequencing for microbiology applications include rapid and reliable high-throughput screening and accurate identification of microbes and microbial genome mutations. Pyrosequencing involves sequencing of DNA by synthesizing the complementary strand a single base at a time, while determining the specific nucleotide being incorporated during the synthesis reaction. The reaction occurs on immobilized single stranded template DNA where the four deoxyribonucleotides (dNTP) are added sequentially and the unincorporated dNTPs are enzymatically degraded before addition of the next dNTP to the synthesis reaction. Detection of the specific base incorporated into the template is monitored by generation of chemiluminescent signals. The order of dNTPs that produce the chemiluminescent signals determines the DNA sequence of the template. The real-time sequencing capability of pyrosequencing technology enables rapid microbial identification in a single assay. In addition, the pyrosequencing instrument, can analyze the full genetic diversity of anti-microbial drug resistance, including typing of SNPs, point mutations, insertions, and deletions, as well as quantification of multiple gene copies that may occur in some anti-microbial resistance patterns.

Pyrosequencing for Rapid Detection of Mycobacterium tuberculosis Resistance to Rifampin, Isoniazid, and Fluoroquinolones ▿

PubMed Central

Bravo, Lulette Tricia C.; Tuohy, Marion J.; Ang, Concepcion; Destura, Raul V.; Mendoza, Myrna; Procop, Gary W.; Gordon, Steven M.; Hall, Geraldine S.; Shrestha, Nabin K.

2009-01-01

After isoniazid and rifampin (rifampicin), the next pivotal drug class in Mycobacterium tuberculosis treatment is the fluoroquinolone class. Mutations in resistance-determining regions (RDR) of the rpoB, katG, and gyrA genes occur with frequencies of 97%, 50%, and 85% among M. tuberculosis isolates resistant to rifampin, isoniazid, and fluoroquinolones, respectively. Sequences are highly conserved, and certain mutations correlate well with phenotypic resistance. We developed a pyrosequencing assay to determine M. tuberculosis genotypic resistance to rifampin, isoniazid, and fluoroquinolones. We characterized 102 M. tuberculosis clinical isolates from the Philippines for susceptibility to rifampin, isoniazid, and ofloxacin by using the conventional submerged-disk proportion method and validated our pyrosequencing assay using these isolates. DNA was extracted and amplified by using PCR primers directed toward the RDR of the rpoB, katG, and gyrA genes, and pyrosequencing was performed on the extracts. The M. tuberculosis H37Rv strain (ATCC 25618) was used as the reference strain. The sensitivities and specificities of pyrosequencing were 96.7% and 97.3%, 63.8% and 100%, and 70.0% and 100% for the detection of resistance to rifampin, isoniazid, and ofloxacin, respectively. Pyrosequencing is thus a rapid and accurate method for detecting M. tuberculosis resistance to these three drugs. PMID:19846642

Pyrosequencing analysis for detection of a BRAFV600E mutation in an FNAB specimen of thyroid nodules.

PubMed

Kim, Suk Kyeong; Kim, Dong-Lim; Han, Hye Seung; Kim, Wan Seop; Kim, Seung Ja; Moon, Won Jin; Oh, Seo Young; Hwang, Tae Sook

2008-06-01

Fine-needle aspiration biopsy (FNAB) is the primary means of distinguishing benign from malignant and of guiding therapeutic intervention in thyroid nodules. However, 10% to 30% of cases with indeterminate cytology in FNAB need other diagnostic tools to refine diagnosis. We compared the pyrosequencing method with the conventional direct DNA sequencing analysis and investigated the usefulness of preoperative BRAF mutation analysis as an adjunct diagnostic tool with routine FNAB. A total of 103 surgically confirmed patients' FNA slides were recruited and DNA was extracted after atypical cells were scraped from the slides. BRAF mutation was analyzed by pyrosequencing and direct DNA sequencing. Sixty-three (77.8%) of 81 histopathologically diagnosed malignant nodules revealed positive BRAF mutation on pyrosequencing analysis. In detail, 63 (84.0%) of 75 papillary thyroid carcinoma (PTC) samples showed positive BRAF mutation, whereas 3 follicular thyroid carcinomas, 1 anaplastic carcinoma, 1 medullary thyroid carcinoma, and 1 metastatic lung carcinoma did not show BRAF mutation. None of 22 benign nodules had BRAF mutation in both pyrosequencing and direct DNA sequencing. Out of 27 thyroid nodules classified as 'indeterminate' on cytologic examination preoperatively, 21 (77.8%) cases turned out to be malignant: 18 PTCs (including 2 follicular variant types) and 3 follicular thyroid carcinomas. Among these, 13 (61.9%) classic PTCs had BRAF mutation. None of 6 benign nodules, including 3 follicular adenomas and 3 nodular hyperplasias, had BRAF mutation. Among 63 PTCs with positive BRAF mutation detected by pyrosequencing analysis, 3 cases did not show BRAF mutation by direct DNA sequencing. Although it was not statistically significant, pyrosequencing was superior to direct DNA sequencing in detecting the BRAF mutation of thyroid nodules (P=0.25). Detecting BRAF mutation by pyrosequencing is more sensitive, faster, and less expensive than direct DNA sequencing and is proposed as an adjunct diagnostic tool in evaluating thyroid nodules of indeterminate cytology.

Qualitative properties of the minimal model of carbon circulation in the biosphere

NASA Astrophysics Data System (ADS)

Pestunov, Aleksandr; Fedotov, Anatoliy; Medvedev, Sergey

2014-05-01

Substantial changes in the biosphere during recent decades have caused legitimate concern in the international community. The fact that feedbacks between the atmospheric CO2 concentration, global temperature, permafrost, ocean CO2 concentration and air humidity increases the risk of catastrophic phenomena on the planetary scale. The precautionary principle allows us to consider greenhouse effect using the mathematical models of the biosphere-climate system. Minimal models do not allow us to make a quantitative description of the "biosphere-climate" system dynamics, which is determined by the aggregate effect of the set of known climatic and biosphere processes. However, the study of such models makes it possible to understand the qualitative mechanisms of biosphere processes and to evaluate their possible consequences. The global minimal model of long-term dynamics of carbon in biosphere is considered basing on assumption that anthropogenous carbon emissions in atmosphere are absent [1]. Qualitative analysis of the model shows that there exists a set of model parameters (taken from the current estimation ranges), such that the system becomes unstable. It is also shown that external influences on the carbon circulation can lead either to degradation of the biosphere or to global temperature change [2]. This work is aimed at revealing the conditions under which the biosphere model can become unstable, which can result in catastrophic changes in the Earth's biogeocenoses. The minimal model of the biosphere-climate system describes an improbable, but, nevertheless, a possible worst-case scenario of the biosphere evolution takes into consideration only the most dangerous biosphere mechanisms and ignores some climate feedbacks (such as transpiration). This work demonstrates the possibility of implementing the trigger mode in the biosphere, which can lead to dramatic changes in the state of the biosphere even without additional burning of fossil fuels. This mode implementation is possible under parameter values of the biosphere, lying within the ranges of their existing estimates. Hence a potential hazard of any drastic change of the biosphere conditions that may speed up possible shift of the biosphere to a new stable state. References 1. Bartsev S.I., Degermendzhi A.G., Fedotov A.M., Medvedev S.B., Pestunov A.I., Pestunov I.A. The Biosphere Trigger Mechanism in the Minimal Model for the Global Carbon Cycle of the Earth // Doklady Earth Sciences, 2012, Vol. 443, Part 2, pp. 489-492. © Pleiades Publishing, Ltd., 2012. 2. Fedotov A.M., Medvedev S.B., Pestunov A.I., Pestunov I.A., Bartsev S.I., Degermendzhi A.G. Qualitative analysis of the minimal model of carbon dynamics in the biosphere // Computational Technologies. 2012. Vol. 17. N 3. pp. 91-108 (in Russian).

Planetary Bootstrap: A Prelude to Biosphere Phenomenology

NASA Astrophysics Data System (ADS)

Kazansky, Alexander B.

2004-08-01

This paper deals with systemic status as well as with some phenomenological and evolutionary aspects of biosphere. Biosphere is represented as multilevel autopoietic system in which different organizational levels are nested into each other. The conceptual model of punctuated epigenesis, biosphere evolutionary process is suggested, in which endogenous planetary organizational crises play role of evolutionary mechanism, creating novelty. The hypothesis is proposed, that the biosphere reaction on the humankind destructive activity reminds the distributed immune response of biological organism, described by F.Varela in his "cognitive immunology". The biosphere evolution is interpreted as the hermeneutical spiral of "Process Being" self-uncovering thus illustrating the historical process of transformation of biosphere as the type of Being in the periods of crises. Some arguments are adduced in favor of biosphere phenomenology development and application of the methods of second-order cybernetics to actual problems of planetary scale.

Theoretical Accuracy of Global Snow-Cover Mapping Using Satellite Data in the Earth Observing System (EOS) Era

NASA Technical Reports Server (NTRS)

Hall, D. K.; Foster, J. L.; Salomonson, V. V.; Klein, A. G.; Chien, J. Y. L.

1998-01-01

Following the launch of the Earth Observing System first morning (EOS-AM1) satellite, daily, global snow-cover mapping will be performed automatically at a spatial resolution of 500 m, cloud-cover permitting, using Moderate Resolution Imaging Spectroradiometer (MODIS) data. A technique to calculate theoretical accuracy of the MODIS-derived snow maps is presented. Field studies demonstrate that under cloud-free conditions when snow cover is complete, snow-mapping errors are small (less than 1%) in all land covers studied except forests where errors are greater and more variable. The theoretical accuracy of MODIS snow-cover maps is largely determined by percent forest cover north of the snowline. Using the 17-class International Geosphere-Biosphere Program (IGBP) land-cover maps of North America and Eurasia, the Northern Hemisphere is classified into seven land-cover classes and water. Snow-mapping errors estimated for each of the seven land-cover classes are extrapolated to the entire Northern Hemisphere for areas north of the average continental snowline for each month. Average monthly errors for the Northern Hemisphere are expected to range from 5 - 10%, and the theoretical accuracy of the future global snow-cover maps is 92% or higher. Error estimates will be refined after the first full year that MODIS data are available.

Using CO2:CO Correlations to Improve Inverse Analyses of Carbon Fluxes

NASA Technical Reports Server (NTRS)

Palmer, Paul I.; Suntharalingam, Parvadha; Jones, Dylan B. A.; Jacob, Daniel J.; Streets, David G.; Fu, Qingyan; Vay, Stephanie A.; Sachse, Glen W.

2006-01-01

Observed correlations between atmospheric concentrations of CO2 and CO represent potentially powerful information for improving CO2 surface flux estimates through coupled CO2-CO inverse analyses. We explore the value of these correlations in improving estimates of regional CO2 fluxes in east Asia by using aircraft observations of CO2 and CO from the TRACE-P campaign over the NW Pacific in March 2001. Our inverse model uses regional CO2 and CO surface fluxes as the state vector, separating biospheric and combustion contributions to CO2. CO2-CO error correlation coefficients are included in the inversion as off-diagonal entries in the a priori and observation error covariance matrices. We derive error correlations in a priori combustion source estimates of CO2 and CO by propagating error estimates of fuel consumption rates and emission factors. However, we find that these correlations are weak because CO source uncertainties are mostly determined by emission factors. Observed correlations between atmospheric CO2 and CO concentrations imply corresponding error correlations in the chemical transport model used as the forward model for the inversion. These error correlations in excess of 0.7, as derived from the TRACE-P data, enable a coupled CO2-CO inversion to achieve significant improvement over a CO2-only inversion for quantifying regional fluxes of CO2.

Gene discovery using massively parallel pyrosequencing to develop ESTs for the flesh fly Sarcophaga crassipalpis

PubMed Central

Hahn, Daniel A; Ragland, Gregory J; Shoemaker, D DeWayne; Denlinger, David L

2009-01-01

Background Flesh flies in the genus Sarcophaga are important models for investigating endocrinology, diapause, cold hardiness, reproduction, and immunity. Despite the prominence of Sarcophaga flesh flies as models for insect physiology and biochemistry, and in forensic studies, little genomic or transcriptomic data are available for members of this genus. We used massively parallel pyrosequencing on the Roche 454-FLX platform to produce a substantial EST dataset for the flesh fly Sarcophaga crassipalpis. To maximize sequence diversity, we pooled RNA extracted from whole bodies of all life stages and normalized the cDNA pool after reverse transcription. Results We obtained 207,110 ESTs with an average read length of 241 bp. These reads assembled into 20,995 contigs and 31,056 singletons. Using BLAST searches of the NR and NT databases we were able to identify 11,757 unique gene elements (E<0.0001) representing approximately 9,000 independent transcripts. Comparison of the distribution of S. crassipalpis unigenes among GO Biological Process functional groups with that of the Drosophila melanogaster transcriptome suggests that our ESTs are broadly representative of the flesh fly transcriptome. Insertion and deletion errors in 454 sequencing present a serious hurdle to comparative transcriptome analysis. Aided by a new approach to correcting for these errors, we performed a comparative analysis of genetic divergence across GO categories among S. crassipalpis, D. melanogaster, and Anopheles gambiae. The results suggest that non-synonymous substitutions occur at similar rates across categories, although genes related to response to stimuli may evolve slightly faster. In addition, we identified over 500 potential microsatellite loci and more than 12,000 SNPs among our ESTs. Conclusion Our data provides the first large-scale EST-project for flesh flies, a much-needed resource for exploring this model species. In addition, we identified a large number of potential microsatellite and SNP markers that could be used in population and systematic studies of S. crassipalpis and other flesh flies. PMID:19454017

Detection of polyoma virus in brain tissue of patients with progressive multifocal leukoencephalopathy by real-time PCR and pyrosequencing.

PubMed

Beck, Rose C; Kohn, Debra J; Tuohy, Marion J; Prayson, Richard A; Yen-Lieberman, Belinda; Procop, Gary W

2004-03-01

We evaluated 2 methods, a LightCycler PCR assay and pyrosequencing for the detection of the JC polyoma virus (JCV) in fixed brain tissue of 10 patients with and 3 control patients without progressive multifocal leukoencephalopathy (PML). Nucleic acid extraction was performed after deparaffinization and proteinase K digestion. The LightCycler assay differentiates the BK virus (BKV), JCV, and SV40 using melt curve analysis. Conventional PCR was used with the same primers to generate products for pyrosequencing. Two sequencing primers were used that differentiate the polyoma viruses. Seven of 11 biopsies (1 patient had 2 biopsies) with PML were positive for JCV by real-time PCR and/or PCR/pyrosequencing. Three of 4 remaining biopsies were positive by real-time PCR but had melting points between JCV and SV40. The 4 specimens that were negative or atypical by LightCycler PCR were positive by traditional PCR, but 1 had an amplicon of lower molecular weight by gel electrophoresis. These were shown to represent JCV by at least 1 of the 2 pyrosequencing primers. The biopsies from patients without PML were PCR negative. Both the LightCycler and pyrosequencing assays are useful for confirming JCV in brain biopsies from patients with PML, but variant JCVs may require supplementary methods to confirm JCV infection.

DOE Office of Scientific and Technical Information (OSTI.GOV)

M. A. Wasiolek

The purpose of this report is to document the biosphere model, the Environmental Radiation Model for Yucca Mountain, Nevada (ERMYN), which describes radionuclide transport processes in the biosphere and associated human exposure that may arise as the result of radionuclide release from the geologic repository at Yucca Mountain. The biosphere model is one of the process models that support the Yucca Mountain Project (YMP) Total System Performance Assessment (TSPA) for the license application (LA), the TSPA-LA. The ERMYN model provides the capability of performing human radiation dose assessments. This report documents the biosphere model, which includes: (1) Describing the referencemore » biosphere, human receptor, exposure scenarios, and primary radionuclides for each exposure scenario (Section 6.1); (2) Developing a biosphere conceptual model using site-specific features, events, and processes (FEPs), the reference biosphere, the human receptor, and assumptions (Section 6.2 and Section 6.3); (3) Building a mathematical model using the biosphere conceptual model and published biosphere models (Sections 6.4 and 6.5); (4) Summarizing input parameters for the mathematical model, including the uncertainty associated with input values (Section 6.6); (5) Identifying improvements in the ERMYN model compared with the model used in previous biosphere modeling (Section 6.7); (6) Constructing an ERMYN implementation tool (model) based on the biosphere mathematical model using GoldSim stochastic simulation software (Sections 6.8 and 6.9); (7) Verifying the ERMYN model by comparing output from the software with hand calculations to ensure that the GoldSim implementation is correct (Section 6.10); and (8) Validating the ERMYN model by corroborating it with published biosphere models; comparing conceptual models, mathematical models, and numerical results (Section 7).« less

Evidence for chlorine recycling - Hydrosphere, biosphere, atmosphere - In a forested wet zone on the Canadian Shield

USGS Publications Warehouse

Milton, G.M.; Milton, J.C.D.; Schiff, S.; Cook, P.; Kotzer, T.G.; Cecil, L.D.

2003-01-01

The ability to measure environmental levels of 36Cl by Accelerator Mass Spectrometry and 3H by 3He-ingrowth Mass Spectrometry has made it possible to use the pulses of these two isotopes released into the atmosphere during nuclear weapons testing as tracers of Cl and water movement in soils and groundwater. The authors have investigated the movement of these tracers below a forested wet zone, and have found that both are retarded to a differing extent in the near surface because of vegetative uptake and recycling. Adsorption by clay particles, followed by slow release to the groundwater, may also be significant. The data accumulated in this region of near vertical recharge have gone a considerable distance towards explaining the anomalously low concentrations of 36Cl measured in the 5 Laurentian Great Lakes, as well as indicating possible mechanisms for large scale Cl- recycling in the atmosphere and biosphere. Identification of the near-term non-conservative behaviour of the Cl- is significant, since such a phenomenon could introduce errors in many watershed calculations, e.g. water residence times, evaporation rates, and mixing calculations. Published by Elsevier Science Ltd.

Comparisons of diazotrophic communities in native and agricultural desert ecosystems reveal plants as important drivers in diversity

PubMed Central

Köberl, Martina; Erlacher, Armin; Ramadan, Elshahat M.; El-Arabi, Tarek F.; Müller, Henry; Bragina, Anastasia; Berg, Gabriele

2015-01-01

Diazotrophs provide the only biological source of fixed atmospheric nitrogen in the biosphere. Although they are the key player for plant-available nitrogen, less is known about their diversity and potential importance in arid ecosystems. We investigated the nitrogenase gene diversity in native and agricultural desert soil as well as within root-associated microbiota of medicinal plants grown in Egypt through the combination of nifH-specific qPCR, fingerprints, amplicon pyrosequencing and fluorescence in situ hybridization–confocal laser scanning microscopy. Although the diazotrophic microbiota were characterized by generally high abundances and diversity, statistically significant differences were found between both soils, the different microhabitats, and between the investigated plants (Matricaria chamomilla L., Calendula officinalis L. and Solanum distichum Schumach. and Thonn.). We observed a considerable community shift from desert to agriculturally used soil that demonstrated a higher abundance and diversity in the agro-ecosystem. The endorhiza was characterized by lower abundances and only a subset of species when compared to the rhizosphere. While the microbiomes of the Asteraceae were similar and dominated by potential root-nodulating rhizobia acquired primarily from soil, the perennial S. distichum generally formed associations with free-living nitrogen fixers. These results underline the importance of diazotrophs in desert ecosystems and additionally identify plants as important drivers in functional gene pool diversity. PMID:26705571

Comparisons of diazotrophic communities in native and agricultural desert ecosystems reveal plants as important drivers in diversity.

PubMed

Köberl, Martina; Erlacher, Armin; Ramadan, Elshahat M; El-Arabi, Tarek F; Müller, Henry; Bragina, Anastasia; Berg, Gabriele

2016-02-01

Diazotrophs provide the only biological source of fixed atmospheric nitrogen in the biosphere. Although they are the key player for plant-available nitrogen, less is known about their diversity and potential importance in arid ecosystems. We investigated the nitrogenase gene diversity in native and agricultural desert soil as well as within root-associated microbiota of medicinal plants grown in Egypt through the combination of nifH-specific qPCR, fingerprints, amplicon pyrosequencing and fluorescence in situ hybridization-confocal laser scanning microscopy. Although the diazotrophic microbiota were characterized by generally high abundances and diversity, statistically significant differences were found between both soils, the different microhabitats, and between the investigated plants (Matricaria chamomilla L., Calendula officinalis L. and Solanum distichum Schumach. and Thonn.). We observed a considerable community shift from desert to agriculturally used soil that demonstrated a higher abundance and diversity in the agro-ecosystem. The endorhiza was characterized by lower abundances and only a subset of species when compared to the rhizosphere. While the microbiomes of the Asteraceae were similar and dominated by potential root-nodulating rhizobia acquired primarily from soil, the perennial S. distichum generally formed associations with free-living nitrogen fixers. These results underline the importance of diazotrophs in desert ecosystems and additionally identify plants as important drivers in functional gene pool diversity. © FEMS 2015.

Density variations and their influence on carbon stocks: case-study on two Biosphere Reserves in the Democratic Republic of Congo

NASA Astrophysics Data System (ADS)

De Ridder, Maaike; De Haulleville, Thalès; Kearsley, Elizabeth; Van den Bulcke, Jan; Van Acker, Joris; Beeckman, Hans

2014-05-01

It is commonly acknowledged that allometric equations for aboveground biomass and carbon stock estimates are improved significantly if density is included as a variable. However, not much attention is given to this variable in terms of exact, measured values and density profiles from pith to bark. Most published case-studies obtain density values from literature sources or databases, this way using large ranges of density values and possible causing significant errors in carbon stock estimates. The use of one single fixed value for density is also not recommended if carbon stock increments are estimated. Therefore, our objective is to measure and analyze a large number of tree species occurring in two Biosphere Reserves (Luki and Yangambi). Nevertheless, the diversity of tree species in these tropical forests is too high to perform this kind of detailed analysis on all tree species (> 200/ha). Therefore, we focus on the most frequently encountered tree species with high abundance (trees/ha) and dominance (basal area/ha) for this study. Increment cores were scanned with a helical X-ray protocol to obtain density profiles from pith to bark. This way, we aim at dividing the tree species with a distinct type of density profile into separate groups. If, e.g., slopes in density values from pith to bark remain stable over larger samples of one tree species, this slope could also be used to correct for errors in carbon (increment) estimates, caused by density values from simplified density measurements or density values from literature. In summary, this is most likely the first study in the Congo Basin that focuses on density patterns in order to check their influence on carbon stocks and differences in carbon stocking based on species composition (density profiles ~ temperament of tree species).

The legacy of biosphere 2 for the study of biospherics and closed ecological systems

NASA Astrophysics Data System (ADS)

Allen, J. P.; Nelson, M.; Alling, A.

The unprecedented challenges of creating Biosphere 2, the world's first laboratory for biospherics, the study of global ecology and long-term closed ecological system dynamics, led to breakthrough developments in many fields, and a deeper understanding of the opportunities and difficulties of material closure. This paper will review accomplishments and challenges, citing some of the key research findings and publications that have resulted from the experiments in Biosphere 2. Engineering accomplishments included development of a technique for variable volume to deal with pressure differences between the facility and outside environment, developing methods of atmospheric leak detection and sealing, while achieving new standards of closure, with an annual atmospheric leakrate of less than 10%, or less than 300 ppm per day. This degree of closure permitted detailed tracking of carbon dioxide, oxygen, and trice gases such as nitrous oxide and ethylene over the seasonal variability of two years. Full closure also necessitated developing new approaches and technologies for complete air, water, and wastewater recycle and reuse within the facility. The development of a soil-based highly productive agricultural system was a first in closed ecological systems, and much was learned about managing a wide variety of crops using non-chemical means of pest and disease control. Closed ecological systems have different temporal biogeochemical cycling and ranges of atmospheric components because of their smaller reservoirs of air, water and soil, and higher concentration of biomass, and Biosphere 2 provided detailed examination and modeling of these accelerated cycles over a period of closure which measured in years. Medical research inside Biosphere 2 included the effects on humans of lowered oxygen: the discovery that human productivity can be maintained with good health with lowered atmospheric oxygen levels could lead to major economies on the design of space stations and planetary/lunar settlements. The improved health resulting from the calorie-restricted but nutrient dense Biosphere 2 diet was the first such scientifically controlled experiment with humans. The success of Biosphere 2 in creating a diversity of terrestrial and marine environments, from rainforest to coral reef, allowed detailed studies with comprehensive measurements such that the dynamics of these complex biomic systems are now better understood. The coral reef ecosystem, the largest artificial reef ever built, catalyzed methods of study now being applied to planetary coral reef systems. Restoration ecology advanced through the creation and study of the dynamics of adaptation and self-organization of the biomes in Biosphere 2. The international interest that Biosphere 2 generated has given new impetus to the public recognition of the sciences of biospheres (biospherics), biomes and closed ecological life systems. The facility, although no longer a materially-closed ecological system, is being used as an educational facility by Columbia University as an introduction to the study of the biosphere and complex system ecology and for carbon dioxide impacts utilizing the complex ecosystems created in Biosphere '.The many lessons learned from Biosphere 2 are being used by its key team of creators in their design and operation of a laboratory-sized closed ecological system, the Laboratory Biosphere, in operation as of March 2002, and for the design of a Mars on Earth ™ prototype life support system for manned missions to Mars and Mars surface habitats. Biosphere 2 is an important foundation for future advances in biospherics and closed ecological system research.

Pyrosequencing for detection of lamivudine-resistant hepatitis B virus.

PubMed

Lindström, Anna; Odeberg, Jacob; Albert, Jan

2004-10-01

Chronic hepatitis B virus (HBV) infection can cause severe liver disease, including cirrhosis and hepatocellular carcinoma. Lamivudine is a relatively recent alternative to alpha interferon for the treatment of HBV infection, but unfortunately, resistance to lamivudine commonly develops during monotherapy. Lamivudine-resistant HBV mutants display specific mutations in the YMDD (tyrosine, methionine, aspartate, aspartate) motif of the viral polymerase (reverse transcriptase [rt]), which is the catalytic site of the enzyme, i.e., methionine 204 to isoleucine (rtM204I) or valine (rtM204V). The latter mutation is often accompanied by a compensatory leucine-to-methionine change at codon 180 (rtL180M). In the present study, a novel sequencing method, pyrosequencing, was applied to the detection of lamivudine resistance mutations and was compared with direct Sanger sequencing. The new pyrosequencing method had advantages in terms of throughput. Experiments with mixtures of wild-type and resistant viruses indicated that pyrosequencing can detect minor sequence variants in heterogeneous virus populations. The new pyrosequencing method was evaluated with a small number of patient samples, and the results showed that the method could be a useful tool for the detection of lamivudine resistance in the clinical setting.

Comparison Study of MS-HRM and Pyrosequencing Techniques for Quantification of APC and CDKN2A Gene Methylation

PubMed Central

Migheli, Francesca; Stoccoro, Andrea; Coppedè, Fabio; Wan Omar, Wan Adnan; Failli, Alessandra; Consolini, Rita; Seccia, Massimo; Spisni, Roberto; Miccoli, Paolo; Mathers, John C.; Migliore, Lucia

2013-01-01

There is increasing interest in the development of cost-effective techniques for the quantification of DNA methylation biomarkers. We analyzed 90 samples of surgically resected colorectal cancer tissues for APC and CDKN2A promoter methylation using methylation sensitive-high resolution melting (MS-HRM) and pyrosequencing. MS-HRM is a less expensive technique compared with pyrosequencing but is usually more limited because it gives a range of methylation estimates rather than a single value. Here, we developed a method for deriving single estimates, rather than a range, of methylation using MS-HRM and compared the values obtained in this way with those obtained using the gold standard quantitative method of pyrosequencing. We derived an interpolation curve using standards of known methylated/unmethylated ratio (0%, 12.5%, 25%, 50%, 75%, and 100% of methylation) to obtain the best estimate of the extent of methylation for each of our samples. We observed similar profiles of methylation and a high correlation coefficient between the two techniques. Overall, our new approach allows MS-HRM to be used as a quantitative assay which provides results which are comparable with those obtained by pyrosequencing. PMID:23326336

Pyrosequencing Analysis of Bench-Scale Nitrifying BiofiltersRemoving Trihalomethanes

EPA Science Inventory

The bacterial biofilm communities in four nitrifying biofilters degrading regulated drinking water trihalomethanes were characterized by 454 pyrosequencing. The three most abundant phylotypes based on total diversity were Nitrosomonas (70%), Nitrobacter (14%), and Chitinophagace...

Implementation of the Biosphere Compatibility Principle in Urban Planning: How to Train Next-Generation Specialists

ERIC Educational Resources Information Center

Ivanova, Zinaida Ilyinichna; Yudenkova, Olga Valeryevna; Ishkov, Aleksandr Dmitrievich; Shnyrenkov, Evgeny Anatolyevich

2015-01-01

The co-authors address the relevant issues concerning the need to implement the principle of the biosphere compatibility as the core prerequisite for the symbiotic co-existence of man and nature. Caring treatment of the biosphere, termination of its excessive exploitation, analysis of the ratio between the biospheric potential of specific areas…

Combining flow cytometry and 16S rRNA gene pyrosequencing: a promising approach for drinking water monitoring and characterization.

PubMed

Prest, E I; El-Chakhtoura, J; Hammes, F; Saikaly, P E; van Loosdrecht, M C M; Vrouwenvelder, J S

2014-10-15

The combination of flow cytometry (FCM) and 16S rRNA gene pyrosequencing data was investigated for the purpose of monitoring and characterizing microbial changes in drinking water distribution systems. High frequency sampling (5 min intervals for 1 h) was performed at the outlet of a treatment plant and at one location in the full-scale distribution network. In total, 52 bulk water samples were analysed with FCM, pyrosequencing and conventional methods (adenosine-triphosphate, ATP; heterotrophic plate count, HPC). FCM and pyrosequencing results individually showed that changes in the microbial community occurred in the water distribution system, which was not detected with conventional monitoring. FCM data showed an increase in the total bacterial cell concentrations (from 345 ± 15 × 10(3) to 425 ± 35 × 10(3) cells mL(-1)) and in the percentage of intact bacterial cells (from 39 ± 3.5% to 53 ± 4.4%) during water distribution. This shift was also observed in the FCM fluorescence fingerprints, which are characteristic of each water sample. A similar shift was detected in the microbial community composition as characterized with pyrosequencing, showing that FCM and genetic fingerprints are congruent. FCM and pyrosequencing data were subsequently combined for the calculation of cell concentration changes for each bacterial phylum. The results revealed an increase in cell concentrations of specific bacterial phyla (e.g., Proteobacteria), along with a decrease in other phyla (e.g., Actinobacteria), which could not be concluded from the two methods individually. The combination of FCM and pyrosequencing methods is a promising approach for future drinking water quality monitoring and for advanced studies on drinking water distribution pipeline ecology. Copyright © 2014 Elsevier Ltd. All rights reserved.

Rapid detection of the CYP2A6*12 hybrid allele by Pyrosequencing technology.

PubMed

Koontz, Deborah A; Huckins, Jacqueline J; Spencer, Antonina; Gallagher, Margaret L

2009-08-24

Identification of CYP2A6 alleles associated with reduced enzyme activity is important in the study of inter-individual differences in drug metabolism. CYP2A6*12 is a hybrid allele that results from unequal crossover between CYP2A6 and CYP2A7 genes. The 5' regulatory region and exons 1-2 are derived from CYP2A7, and exons 3-9 are derived from CYP2A6. Conventional methods for detection of CYP2A6*12 consist of two-step PCR protocols that are laborious and unsuitable for high-throughput genotyping. We developed a rapid and accurate method to detect the CYP2A6*12 allele by Pyrosequencing technology. A single set of PCR primers was designed to specifically amplify both the CYP2A6*1 wild-type allele and the CYP2A6*12 hybrid allele. An internal Pyrosequencing primer was used to generate allele-specific sequence information, which detected homozygous wild-type, heterozygous hybrid, and homozygous hybrid alleles. We first validated the assay on 104 DNA samples that were also genotyped by conventional two-step PCR and by cycle sequencing. CYP2A6*12 allele frequencies were then determined using the Pyrosequencing assay on 181 multi-ethnic DNA samples from subjects of African American, European Caucasian, Pacific Rim, and Hispanic descent. Finally, we streamlined the Pyrosequencing assay by integrating liquid handling robotics into the workflow. Pyrosequencing results demonstrated 100% concordance with conventional two-step PCR and cycle sequencing methods. Allele frequency data showed slightly higher prevalence of the CYP2A6*12 allele in European Caucasians and Hispanics. This Pyrosequencing assay proved to be a simple, rapid, and accurate alternative to conventional methods, which can be easily adapted to the needs of higher-throughput studies.

Performance of Different Analytical Software Packages in Quantification of DNA Methylation by Pyrosequencing.

PubMed

Grasso, Chiara; Trevisan, Morena; Fiano, Valentina; Tarallo, Valentina; De Marco, Laura; Sacerdote, Carlotta; Richiardi, Lorenzo; Merletti, Franco; Gillio-Tos, Anna

2016-01-01

Pyrosequencing has emerged as an alternative method of nucleic acid sequencing, well suited for many applications which aim to characterize single nucleotide polymorphisms, mutations, microbial types and CpG methylation in the target DNA. The commercially available pyrosequencing systems can harbor two different types of software which allow analysis in AQ or CpG mode, respectively, both widely employed for DNA methylation analysis. Aim of the study was to assess the performance for DNA methylation analysis at CpG sites of the two pyrosequencing software which allow analysis in AQ or CpG mode, respectively. Despite CpG mode having been specifically generated for CpG methylation quantification, many investigations on this topic have been carried out with AQ mode. As proof of equivalent performance of the two software for this type of analysis is not available, the focus of this paper was to evaluate if the two modes currently used for CpG methylation assessment by pyrosequencing may give overlapping results. We compared the performance of the two software in quantifying DNA methylation in the promoter of selected genes (GSTP1, MGMT, LINE-1) by testing two case series which include DNA from paraffin embedded prostate cancer tissues (PC study, N = 36) and DNA from blood fractions of healthy people (DD study, N = 28), respectively. We found discrepancy in the two pyrosequencing software-based quality assignment of DNA methylation assays. Compared to the software for analysis in the AQ mode, less permissive criteria are supported by the Pyro Q-CpG software, which enables analysis in CpG mode. CpG mode warns the operators about potential unsatisfactory performance of the assay and ensures a more accurate quantitative evaluation of DNA methylation at CpG sites. The implementation of CpG mode is strongly advisable in order to improve the reliability of the methylation analysis results achievable by pyrosequencing.

Multiplex pyrosequencing method to determine CYP2C9*3, VKORC1*2, and CYP4F2*3 polymorphisms simultaneously: its application to a Korean population and comparisons with other ethnic groups.

PubMed

Kim, Kyoung-Ah; Song, Wan-Geun; Lee, Hae-Mi; Joo, Hyun-Jin; Park, Ji-Young

2014-11-01

Warfarin is an anticoagulant that is difficult to administer because of the wide variation in dose requirements to achieve a therapeutic effect. CYP2C9, VKROC1, and CYP4F2 play important roles in warfarin metabolism, and their genetic polymorphisms are related to the variability in dose determination. In this study we describe a new multiplex pyrosequencing method to identify CYP2C9*3 (rs1057910), VKORC1*2 (rs9923231), and CYP4F2*3 (rs2108661) simultaneously. A multiplex pyrosequencing method to simultaneously detect CYP2C9*3, VKORC1*2, and CYP4F2*3 alleles was designed. We assessed the allele frequencies of the polymorphisms in 250 Korean subjects using the multiplex pyrosequencing method. The results showed 100 % concordance between single and multiplex pyrosequencing methods, and the polymorphisms identified by pyrosequencing were also validated with the direct sequencing method. The allele frequencies of these polymorphisms in this population were as follows: 0.040 for CYP2C9*3, 0.918 for VKORC1*2, and 0.416 for CYP4F2*3. Although the allele frequencies of the CYP2C9*3 and VKROC1*2 were comparable to those in Japanese and Chinese populations, their frequencies in this Korean population differed from those in other ethnic groups; the CYP4F2*3 frequency was the highest among other ethnic populations including Chinese and Japanese populations. The pyrosequencing methods developed were rapid and reliable for detecting CYP2C9*3, VKORC1*2, and CYP4F2*3. Large ethnic differences in the frequency of these genetic polymorphisms were noted among ethnic groups. CYP4F2*3 exhibited its highest allele frequency among other ethnic populations compared to that in a Korean population.

Biosphere reserves: Attributes for success.

PubMed

Van Cuong, Chu; Dart, Peter; Hockings, Marc

2017-03-01

Biosphere reserves established under the UNESCO Man and the Biosphere Program aim to harmonise biodiversity conservation and sustainable development. Concerns over the extent to which the reserve network was living up to this ideal led to the development of a new strategy in 1995 (the Seville Strategy) to enhance the operation of the network of reserves. An evaluation of effectiveness of management of the biosphere reserve network was called for as part of this strategy. Expert opinion was assembled through a Delphi Process to identify successful and less successful reserves and investigate common factors influencing success or failure. Ninety biosphere reserves including sixty successful and thirty less successful reserves in 42 countries across all five Man and the Biosphere Program regions were identified. Most successful sites are the post-Seville generation while the majority of unsuccessful sites are pre-Seville that are managed as national parks and have not been amended to conform to the characteristics that are meant to define a biosphere reserve. Stakeholder participation and collaboration, governance, finance and resources, management, and awareness and communication are the most influential factors in the success or failure of the biosphere reserves. For success, the biosphere reserve concept needs to be clearly understood and applied through landscape zoning. Designated reserves then need a management system with inclusive good governance, strong participation and collaboration, adequate finance and human resource allocation and stable and responsible management and implementation. All rather obvious but it is difficult to achieve without commitment to the biosphere reserve concept by the governance authorities. Copyright © 2016 Elsevier Ltd. All rights reserved.

The Impact of Prior Biosphere Models in the Inversion of Global Terrestrial CO2 Fluxes by Assimilating OCO-2 Retrievals

NASA Technical Reports Server (NTRS)

Philip, Sajeev; Johnson, Matthew S.

2018-01-01

Atmospheric mixing ratios of carbon dioxide (CO2) are largely controlled by anthropogenic emissions and biospheric fluxes. The processes controlling terrestrial biosphere-atmosphere carbon exchange are currently not fully understood, resulting in terrestrial biospheric models having significant differences in the quantification of biospheric CO2 fluxes. Atmospheric transport models assimilating measured (in situ or space-borne) CO2 concentrations to estimate "top-down" fluxes, generally use these biospheric CO2 fluxes as a priori information. Most of the flux inversion estimates result in substantially different spatio-temporal posteriori estimates of regional and global biospheric CO2 fluxes. The Orbiting Carbon Observatory 2 (OCO-2) satellite mission dedicated to accurately measure column CO2 (XCO2) allows for an improved understanding of global biospheric CO2 fluxes. OCO-2 provides much-needed CO2 observations in data-limited regions facilitating better global and regional estimates of "top-down" CO2 fluxes through inversion model simulations. The specific objectives of our research are to: 1) conduct GEOS-Chem 4D-Var assimilation of OCO-2 observations, using several state-of-the-science biospheric CO2 flux models as a priori information, to better constrain terrestrial CO2 fluxes, and 2) quantify the impact of different biospheric model prior fluxes on OCO-2-assimilated a posteriori CO2 flux estimates. Here we present our assessment of the importance of these a priori fluxes by conducting Observing System Simulation Experiments (OSSE) using simulated OCO-2 observations with known "true" fluxes.

Reviewing Biosphere Reserves globally: effective conservation action or bureaucratic label?

PubMed

Coetzer, Kaera L; Witkowski, Edward T F; Erasmus, Barend F N

2014-02-01

The Biosphere Reserve (BR) model of UNESCO's Man and the Biosphere Programme reflects a shift towards more accountable conservation. Biosphere Reserves attempt to reconcile environmental protection with sustainable development; they explicitly acknowledge humans, and human interests in the conservation landscape while still maintaining the ecological values of existing protected areas. Conceptually, this model is attractive, with 610 sites currently designated globally. Yet the practical reality of implementing dual 'conservation' and 'development' goals is challenging, with few examples successfully conforming to the model's full criteria. Here, we review the history of Biosphere Reserves from first inception in 1974 to the current status quo, and examine the suitability of the designation as an effective conservation model. We track the spatial expansion of Biosphere Reserves globally, assessing the influence of the Statutory Framework of the World Network of Biosphere Reserves and Seville strategy in 1995, when the BR concept refocused its core objectives on sustainable development. We use a comprehensive range of case studies to discuss conformity to the Programme, the social and ecological consequences associated with implementation of the designation, and challenges in aligning conservation and development. Given that the 'Biosphere Reserve' label is a relatively unknown designation in the public arena, this review also provides details on popularising the Biosphere Reserve brand, as well as prospects for further research, currently unexploited, but implicit in the designation. © 2013 The Authors. Biological Reviews © 2013 Cambridge Philosophical Society.

Anthropogenic transformation of the terrestrial biosphere.

PubMed

Ellis, Erle C

2011-03-13

Human populations and their use of land have transformed most of the terrestrial biosphere into anthropogenic biomes (anthromes), causing a variety of novel ecological patterns and processes to emerge. To assess whether human populations and their use of land have directly altered the terrestrial biosphere sufficiently to indicate that the Earth system has entered a new geological epoch, spatially explicit global estimates of human populations and their use of land were analysed across the Holocene for their potential to induce irreversible novel transformation of the terrestrial biosphere. Human alteration of the terrestrial biosphere has been significant for more than 8000 years. However, only in the past century has the majority of the terrestrial biosphere been transformed into intensively used anthromes with predominantly novel anthropogenic ecological processes. At present, even were human populations to decline substantially or use of land become far more efficient, the current global extent, duration, type and intensity of human transformation of ecosystems have already irreversibly altered the terrestrial biosphere at levels sufficient to leave an unambiguous geological record differing substantially from that of the Holocene or any prior epoch. It remains to be seen whether the anthropogenic biosphere will be sustained and continue to evolve.

[Search for life in deep biospheres].

PubMed

Naganuma, Takeshi

2003-12-01

The life in deep biospheres bridges conventional biology and future exobiology. This review focuses the microbiological studies from the selected deep biospheres, i.e., deep-sea hydrothermal vents, sub-hydrothermal vents, terrestrial subsurface and a sub-glacier lake. The dark biospheres facilitate the emergence of organisms and communities dependent on chemolithoautotrophy, which are overwhelmed by photoautotrophy (photosynthesis) in the surface biospheres. The life at deep-sea hydrothermal vents owes much to chemolithoautotrophy based on the oxidation of sulfide emitted from the vents. It is likely that similarly active bodies such as the Jovian satellite Europa may have hydrothermal vents and associated biological communities. Anoxic or anaerobic condition is characteristic of deep subsurface biospheres. Subsurface microorganisms exploit available oxidants, or terminal electron acceptors (TEA), for anaerobic respiration. Sulfate, nitrate, iron (III) and CO2 are the representative TEAs in the deep subsurface. Below the 3000-4000 m-thick glacier on Antarctica, there have been >70 lakes with liquid water located. One of such sub-glacial lakes, Lake Vostok, is about to be drill-penetrated for microbiological studies. These deep biosphere "platforms" provide new knowledge about the diversity and potential of the Earth's life. The expertise obtained from the deep biosphere expeditions will facilitate the capability of exobiologial exploration.

Application of Biosphere Compatibility Indicator for Assessment of the Effectiveness of Environmental Protection Methods

NASA Astrophysics Data System (ADS)

Bakaeva, N. V.; Vorobyov, S. A.; Chernyaeva, I. V.

2017-11-01

The article is devoted to the issue of using the biosphere compatibility indicator to assess the effectiveness of environmental protection methods. The indicator biosphere compatibility was proposed by the vice-president of RAASN (Russian Academy of Architecture and Building Sciences), Doctor of Technical Sciences, Professor V.I. Ilyichev. This indicator allows one to assess not only qualitatively but also quantitatively the degree of urban areas development from the standpoint of preserving the biosphere in urban ecosystems while performing the city’s main functions. The integral biosphere compatibility indicator allows us to assess not only the current ecological situation in the territory under consideration but also to plan the forecast of its changes for the new construction projects implementation or for the reconstruction of the existing ones. The biosphere compatibility indicator, which is a mathematical expression of the tripartite balance (technosphere, biosphere and population of this area), allows us to quantify the effectiveness degree of different methods for environment protection to choose the most effective one under these conditions.

Feedbacks between climate change and biosphere integrity

NASA Astrophysics Data System (ADS)

Lade, Steven; Anderies, J. Marty; Donges, Jonathan; Steffen, Will; Rockström, Johan; Richardson, Katherine; Cornell, Sarah; Norberg, Jon; Fetzer, Ingo

2017-04-01

The terrestrial and marine biospheres sink substantial fractions of human fossil fuel emissions. How the biosphere's capacity to sink carbon depends on biodiversity and other measures of biosphere integrity is however poorly understood. Here, we (1): review assumptions from literature regarding the relationships between the carbon cycle and the terrestrial and marine biospheres; and (2) explore the consequences of these different assumptions for climate feedbacks using the stylised carbon cycle model PB-INT. We find that: terrestrial biodiversity loss could significantly dampen climate-carbon cycle feedbacks; direct biodiversity effects, if they exist, could rival temperature increases from low-emission trajectories; and the response of the marine biosphere is critical for longer term climate change. Simple, low-dimensional climate models such as PB-INT can help assess the importance of still unknown or controversial earth system processes such as biodiversity loss for climate feedbacks. This study constitutes the first detailed study of the interactions between climate change and biosphere integrity, two of the 'planetary boundaries'.

Biosphere Reserve for All: Potentials for Involving Underrepresented Age Groups in the Development of a Biosphere Reserve through Intergenerational Practice.

PubMed

Mitrofanenko, Tamara; Snajdr, Julia; Muhar, Andreas; Penker, Marianne; Schauppenlehner-Kloyber, Elisabeth

2018-05-22

Stakeholder participation is of high importance in UNESCO biosphere reserves as model regions for sustainable development; however, certain groups remain underrepresented. The paper proposes Intergenerational Practice (IP) as a means of involving youth and elderly women and explores its options and barriers, using the example of the Salzburger Lungau and Kärntner Nockberge Biosphere Reserve in Austria. Case study analysis is used involving mixed methods. The results reveal obstacles and motivations to participating in biosphere reserve implementation and intergenerational activities for the youth and the elderly women and imply that much potential for IP exists in the biosphere reserve region. The authors propose suitable solutions from the intergenerational field to overcome identified participation obstacles and suggest benefits of incorporating IP as a management tool into biosphere reserve activities. Suggestions for future research include evaluating applications of IP in the context of protected areas, testing of methods used in other contexts, and contribution to theory development.

Investigating bacterial populations in styrene-degrading biofilters by 16S rDNA tag pyrosequencing.

PubMed

Portune, Kevin J; Pérez, M Carmen; Álvarez-Hornos, F Javier; Gabaldón, Carmen

2015-01-01

Microbial biofilms are essential components in the elimination of pollutants within biofilters, yet still little is known regarding the complex relationships between microbial community structure and biodegradation function within these engineered ecosystems. To further explore this relationship, 16S rDNA tag pyrosequencing was applied to samples taken at four time points from a styrene-degrading biofilter undergoing variable operating conditions. Changes in microbial structure were observed between different stages of biofilter operation, and the level of styrene concentration was revealed to be a critical factor affecting these changes. Bacterial genera Azoarcus and Pseudomonas were among the dominant classified genera in the biofilter. Canonical correspondence analysis (CCA) and correlation analysis revealed that the genera Brevundimonas, Hydrogenophaga, and Achromobacter may play important roles in styrene degradation under increasing styrene concentrations. No significant correlations (P > 0.05) could be detected between biofilter operational/functional parameters and biodiversity measurements, although biological heterogeneity within biofilms and/or technical variability within pyrosequencing may have considerably affected these results. Percentages of selected bacterial taxonomic groups detected by fluorescence in situ hybridization (FISH) were compared to results from pyrosequencing in order to assess the effectiveness and limitations of each method for identifying each microbial taxon. Comparison of results revealed discrepancies between the two methods in the detected percentages of numerous taxonomic groups. Biases and technical limitations of both FISH and pyrosequencing, such as the binding of FISH probes to non-target microbial groups and lack of classification of sequences for defined taxonomic groups from pyrosequencing, may partially explain some differences between the two methods.

Carbon Monitoring System Flux Estimation and Attribution: Impact of ACOS-GOSAT X(CO2) Sampling on the Inference of Terrestrial Biospheric Sources and Sinks

NASA Technical Reports Server (NTRS)

Liu, Junjie; Bowman, Kevin W.; Lee, Memong; Henze, David K.; Bousserez, Nicolas; Brix, Holger; Collatz, G. James; Menemenlis, Dimitris; Ott, Lesley; Pawson, Steven;

Looking For a Needle in the Haystack: Deciphering Indigenous 1.79 km Deep Subsurface Microbial Communities from Drilling Mud Contaminants Using 454 Pyrotag Sequencing

NASA Astrophysics Data System (ADS)

Dong, Y.; Cann, I.; Mackie, R.; Price, N.; Flynn, T. M.; Sanford, R.; Miller, P.; Chia, N.; Kumar, C. G.; Kim, P.; Sivaguru, M.; Fouke, B. W.

2010-12-01

Knowledge of the composition, structure and activity of microbial communities that live in deeply buried sedimentary rocks is fundamental to the future of subsurface biosphere stewardship as it relates to hydrocarbon exploration and extraction, carbon sequestration, gas storage and groundwater management. However, the study of indigenous subsurface microorganisms has been limited by the technical challenges of collecting deep formation water samples that have not been heavily contaminated by the mud used to drill the wells. To address this issue, a “clean-sampling method” deploying the newly developed Schlumberger Quicksilver MDT probe was used to collect a subsurface sample at a depth of 1.79 km (5872 ft) from an exploratory well within Cambrian-age sandstones in the Illinois Basin. This yielded a formation water sample that was determined to have less than 4% drilling mud contamination based on tracking changes in the aqueous geochemistry of the formation water during ~3 hours of pumping at depth prior to sample collection. A suite of microscopy and culture-independent molecular analyses were completed using the DNA extracted from microbial cells in the formation water, which included 454 amplicon pyrosequencing that targeted the V1-V3 hypervariable region of bacterial 16S rRNA gene sequences. Results demonstrated an extremely low diversity microbial community living in formation water at 1.79 km-depth. More than 95 % of the total V1-V3 pyrosequencing reads (n=11574) obtained from the formation water were affiliated with a halophilic γ-proteobacterium and most closely related to the genus Halomonas. In contrast, about 3 % of the V1-V3 sequences in the drilling mud library (n=13044) were classified as genus Halomonas but were distinctly different and distantly related to the formation water Halomonas detected at 1.79 km-depth. These results were consistent with those obtained using a suite of other molecular screens (e.g., Terminal-Restriction Fragment Length Polymorphism (T-RFLP) and the initial full length 16S rRNA amplicon libraries) and bioinformatic analyses (e.g., 16S rRNA and Open Reading Frame (ORF) calls established from the 454 metagenomic community analyses). Functional pathway modeling is underway to evaluate the adaptation of this indigenous microbial community to the hydrologic and geologic history of the deep subsurface environment of the Illinois Basin.

Rapid detection method for Bacillus anthracis using a combination of multiplexed real-time PCR and pyrosequencing and its application for food biodefense.

PubMed

Janzen, Timothy W; Thomas, Matthew C; Goji, Noriko; Shields, Michael J; Hahn, Kristen R; Amoako, Kingsley K

2015-02-01

Bacillus anthracis, the causative agent of anthrax, has the capacity to form highly resilient spores as part of its life cycle. The potential for the dissemination of these spores using food as a vehicle is a huge public health concern and, hence, requires the development of a foodborne bioterrorism response approach. In this work, we address a critical gap in food biodefense by presenting a novel, combined, sequential method involving the use of real-time PCR and pyrosequencing for the rapid, specific detection of B. anthracis spores in three food matrices: milk, apple juice, and bottled water. The food samples were experimentally inoculated with 40 CFU ml(-1), and DNA was extracted from the spores and analyzed after immunomagnetic separation. Applying the combination of multiplex real-time PCR and pyrosequencing, we successfully detected the presence of targets on both of the virulence plasmids and the chromosome. The results showed that DNA amplicons generated from a five-target multiplexed real-time PCR detection using biotin-labeled primers can be used for single-plex pyrosequencing detection. The combined use of multiplexed real-time PCR and pyrosequencing is a novel, rapid detection method for B. anthracis from food and provides a tool for accurate, quantitative identification with potential biodefense applications.

Southern Appalachian Man and the Biosphere - SAMAB.ORG

Science.gov Websites

Southern Appalachian Man and the Biosphere Home Who We Are SAMAB Initiatives Archives Welcome to SAMAB This slideshow requires JavaScript. The Southern Appalachian Man and the Biosphere (SAMAB

Ciliates and the rare biosphere-community ecology and population dynamics.

PubMed

Weisse, Thomas

2014-01-01

Application of deep sequencing technologies to environmental samples and some detailed morphological studies suggest that there is a vast, yet unexplored rare ciliate biosphere, tentatively defined in terms of operational taxonomic units. However, very few studies complemented molecular and phylogenetic data with morphological and ecological descriptions of the species inventory. This is mainly because the sampling effort increases strongly with decreasing species abundance. In spite of this limited knowledge, it is clear that species that are rare under certain environmental conditions (temporal rare biosphere) may become abundant when the physical, chemical, and biological variables of their habitat change. Furthermore, some species may always be present in low numbers if their dispersal rates are exceedingly high (accidental rare biosphere). An intriguing question is whether there are some species that are always rare, i.e., in every suitable environment. This permanent rare biosphere is conceptually different from the temporal rare biosphere. This review characterizes typical aquatic habitats of the rare ciliate biosphere, portrays different scenarios under which some or even many species may be permanently rare (background fauna), and identifies some fundamental questions that need to be addressed to achieve a better understanding of the population dynamics of the rare ciliate biosphere. © 2014 The Authors The Journal of Eukaryotic Microbiology published by Wiley Periodicals, Inc. on behalf of International Society of Protistologists.

Regionally strong feedbacks between the atmosphere and terrestrial biosphere

NASA Astrophysics Data System (ADS)

Green, Julia K.; Konings, Alexandra G.; Alemohammad, Seyed Hamed; Berry, Joseph; Entekhabi, Dara; Kolassa, Jana; Lee, Jung-Eun; Gentine, Pierre

2017-06-01

The terrestrial biosphere and atmosphere interact through a series of feedback loops. Variability in terrestrial vegetation growth and phenology can modulate fluxes of water and energy to the atmosphere, and thus affect the climatic conditions that in turn regulate vegetation dynamics. Here we analyse satellite observations of solar-induced fluorescence, precipitation, and radiation using a multivariate statistical technique. We find that biosphere-atmosphere feedbacks are globally widespread and regionally strong: they explain up to 30% of precipitation and surface radiation variance in regions where feedbacks occur. Substantial biosphere-precipitation feedbacks are often found in regions that are transitional between energy and water limitation, such as semi-arid or monsoonal regions. Substantial biosphere-radiation feedbacks are often present in several moderately wet regions and in the Mediterranean, where precipitation and radiation increase vegetation growth. Enhancement of latent and sensible heat transfer from vegetation accompanies this growth, which increases boundary layer height and convection, affecting cloudiness, and consequently incident surface radiation. Enhanced evapotranspiration can increase moist convection, leading to increased precipitation. Earth system models underestimate these precipitation and radiation feedbacks mainly because they underestimate the biosphere response to radiation and water availability. We conclude that biosphere-atmosphere feedbacks cluster in specific climatic regions that help determine the net CO2 balance of the biosphere.

Regionally Strong Feedbacks Between the Atmosphere and Terrestrial Biosphere

NASA Technical Reports Server (NTRS)

Green, Julia K.; Konings, Alexandra G.; Alemohammad, Seyed Hamed; Lee, Jung-Eun; Berry, Joseph; Entekhabi, Dara; Kolassa, Jana; Gentine, Pierre

2017-01-01

The terrestrial biosphere and atmosphere interact through a series of feedback loops. Variability in terrestrial vegetation growth and phenology can modulate fluxes of water and energy to the atmosphere, and thus affect the climatic conditions that in turn regulate vegetation dynamics. Here we analyze satellite observations of solar-induced fluorescence, precipitation, and radiation using a multivariate statistical technique. We find that biosphere-atmosphere feedbacks are globally widespread and regionally strong: they explain up to 30 of precipitation and surface radiation variance. Substantial biosphere-precipitation feedbacks are often found in regions that are transitional between energy and water limitation, such as semi-arid or monsoonal regions. Substantial biosphere-radiation feedbacks are often present in several moderately wet regions and in the Mediterranean, where precipitation and radiation increase vegetation growth. Enhancement of latent and sensible heat transfer from vegetation accompanies this growth, which increases boundary layer height and convection, affecting cloudiness, and consequently incident surface radiation. Enhanced evapotranspiration can increase moist convection, leading to increased precipitation. Earth system models underestimate these precipitation and radiation feedbacks mainly because they underestimate the biosphere response to radiation and water availability. We conclude that biosphere-atmosphere feedbacks cluster in specific climatic regions that help determine the net CO2 balance of the biosphere.

Historical overview of the Biosphere 2 project

NASA Technical Reports Server (NTRS)

Allen, John P.

1990-01-01

An extensive historical overview is given of the Biosphere 2 project. The overview begins in late 1969, as the moon landings commenced, when work began on ecological projects which laid the conceptual foundation for the current Biosphere 2 project. Continuing through to taking a complete functional suite of microbes together with their associated aquatic elements and an air volume and putting them inside a closed lab flask in which to measure the oxygen and CO2 levels, study energy flows and visually observe the changes therein. The laws of biospherics formulated by the author which can be tested in the Biosphere 2 project are listed.

Interpretation of Biosphere Reserves.

ERIC Educational Resources Information Center

Merriman, Tim

1994-01-01

Introduces the Man and the Biosphere Programme (MAB) to monitor the 193 biogeographical provinces of the Earth and the creation of biosphere reserves. Highlights the need for interpreters to become familiar or involved with MAB program activities. (LZ)

Biomedical program at Space Biospheres Ventures

NASA Technical Reports Server (NTRS)

Walford, Roy

1990-01-01

There are many similarities and some important differences between potential health problems of Biosphere 2 and those of which might be anticipated for a space station or a major outpost on Mars. The demands of time, expense, and equipment would not readily allow medical evacuation from deep space for a serious illness or major trauma, whereas personnel can easily be evacuated from Biosphere 2 if necessary. Treatment facilities can be somewhat less inclusive, since distance would not compel the undertaking of heroic measures or highly complicated surgical procedures on site, and with personnel not fully trained for these procedures. The similarities are given between medical requirements of Biosphere 2 and the complex closed ecological systems of biospheres in space or on Mars. The major problems common to all these would seem to be trauma, infection, and toxicity. It is planned that minor and moderate degrees of trauma, including debridement and suturing of wounds, x ray study of fractures, will be done within Biosphere 2. Bacteriologic and fungal infections, and possibly allergies to pollen or spores are expected to be the commonest medical problem within Biosphere 2.

Use of the method of biosphere compatibility for the assessment of environmental protection methods

NASA Astrophysics Data System (ADS)

Vorobyov, Sergey

2018-01-01

The article is devoted to the question of using the indicator of biosphere compatibility for assessing the effectiveness of environmental protection methods. The indicator of biosphere compatibility was proposed by the vice-president of RAASN (Russian Academy of Architecture and Building Sciences), Doctor of Technical Sciences, Professor V.I. Ilyichev. This indicator is allows not only qualitatively but also quantitatively to assess the degree of development of urban urban areas, from the standpoint of preserving the biosphere in urban ecosystems while realizing the city’s main functions. The integral indicator of biosphere compatibility is allows us to assess not only the current ecological situation in the territory under consideration, but also to plan the forecast of its changes for building the new construction projects, or for reconstructing existing ones. The indicator of biosphere compatibility, which is a mathematical expression of the tripartite balance (technosphere, biosphere and population of this area), is allows us to quantify the degree of effectiveness of different method of protecting the environment for choose the most effective for these conditions.

The Legacy of Biosphere 2 for Biospherics and Closed Ecological System Research

NASA Astrophysics Data System (ADS)

Allen, J.; Alling, A.; Nelson, M.

The unprecedented challenges of creating Biosphere 2, the world's first laboratory for biospherics, the study of global ecology and long-term closed ecological system dynamics led to breakthrough developments in many fields, and a deeper understanding of the opportunities and difficulties of material closure. This paper will review these accomplishments and challenges, citing some of the key research accomplishments and publications which have resulted from the experiments in Biosphere 2. Engineering accomplishments included development of a technique for variable volume to deal with pressure differences between the facility and outside environment, developing methods of leak detection and sealing, and achieving new standards of closure, with an annual atmospheric leakrate of less than 10%, or less than 300 ppm per day. This degree of closure permitted detailed tracking of carbon dioxide, oxygen, and trace gases such as nitrous oxide and ethylene over the seasonal variability of two years. Full closure also necessitated developing new approaches and technologies for complete air, water, and wastewater recycle and reuse within the facility. The development of a soil-based highly productive agricultural system was a first in closed ecological systems, and much was learned about managing a wide variety of crops using non-chemical means of pest and disease control. Closed ecological systems have different temporal b ogeochemical cycling and ranges ofi atmospheric components because of their smaller reservoirs of air, water and soil, and higher concentration of biomass, and Biosphere 2 provided detailed examination and modeling of these accelerated cycles over a period of closure which measured in years. Medical research inside Biosphere 2 included the effects on humans of lowered oxygen: the discovery that human productivity can be maintained down to 15% oxygen could lead to major economies on the design of space stations and planetary/lunar settlements. The improved health resulting from the calorie-restricted but nutrient dense Biosphere 2 diet was the first such scientifically-controlled experiment with humans. The success of Biosphere in creating a diversity of terrestrial and marine environments, from rainforest to coral reef, allowed detailed studies with comprehensive measurements such that the dynamics of these complex biomic systems can be better understood. The coral reef ecosystem, the largest artificial reef ever built, catalyzed methods of study now being applied to planetary coral reef systems. Restoration ecology can learn much from the creation and dynamics of adaptation of the biomes in Biosphere 2. The international interest that Biosphere 2 generated has given new impetus to the public recognition of the sciences of biospheres, biospherics, biomes and closed ecological life systems. The facility is still being used as an educational facility by Columbia University as an introduction to the study of the biosphere and complex system ecology. The many lessons learned from Biosphere 2, from its successes, surprises and challenges, is being used by its key team of creators as the foundations for their design of a laboratory-sized closed ecological system and Mars on Earth prototype life support system for Mars exploration and can be an important foundation for future advances in biospherics and closed ecological system research.

Evaluation of culture-based techniques and 454 pyrosequencing for the analysis of fungal diversity in potting media and organic fertilizers.

PubMed

Al-Sadi, A M; Al-Mazroui, S S; Phillips, A J L

2015-08-01

Potting media and organic fertilizers (OFs) are commonly used in agricultural systems. However, there is a lack of studies on the efficiency of culture-based techniques in assessing the level of fungal diversity in these products. A study was conducted to investigate the efficiency of seven culture-based techniques and pyrosequencing for characterizing fungal diversity in potting media and OFs. Fungal diversity was evaluated using serial dilution, direct plating and baiting with carrot slices, potato slices, radish seeds, cucumber seeds and cucumber cotyledons. Identity of all the isolates was confirmed on the basis of the internal transcribed spacer region of the ribosomal RNA (ITS rRNA) sequence data. The direct plating technique was found to be superior over other culture-based techniques in the number of fungal species detected. It was also found to be simple and the least time consuming technique. Comparing the efficiency of direct plating with 454 pyrosequencing revealed that pyrosequencing detected 12 and 15 times more fungal species from potting media and OFs respectively. Analysis revealed that there were differences between potting media and OFs in the dominant phyla, classes, orders, families, genera and species detected. Zygomycota (52%) and Chytridiomycota (60%) were the predominant phyla in potting media and OFs respectively. The superiority of pyrosequencing over cultural methods could be related to the ability to detect obligate fungi, slow growing fungi and fungi that exist at low population densities. The evaluated methods in this study, especially direct plating and pyrosequencing, may be used as tools to help detect and reduce movement of unwanted fungi between countries and regions. © 2015 The Society for Applied Microbiology.

The diversity and structure of marine protists in the coastal waters of China revealed by morphological observation and 454 pyrosequencing

NASA Astrophysics Data System (ADS)

Liu, Yun; Song, Shuqun; Chen, Tiantian; Li, Caiwen

2017-04-01

Pyrosequencing of the 18S rRNA gene has been widely adopted to study the eukaryotic diversity in various types of environments, and has an advantage over traditional morphology methods in exploring unknown microbial communities. To comprehensively assess the diversity and community composition of marine protists in the coastal waters of China, we applied both morphological observations and high-throughput sequencing of the V2 and V3 regions of 18S rDNA simultaneously to analyze samples collected from the surface layer of the Yellow and East China Seas. Dinoflagellates, diatoms and ciliates were the three dominant protistan groups as revealed by the two methods. Diatoms were the first dominant protistan group in the microscopic observations, with Skeletonema mainly distributed in the nearshore eutrophic waters and Chaetoceros in higher temperature and higher pH waters. The mixotrophic dinoflagellates, Gymnodinium and Gyrodinium, were more competitive in the oligotrophic waters. The pyrosequencing method revealed an extensive diversity of dinoflagellates. Chaetoceros was the only dominant diatom group in the pyrosequencing dataset. Gyrodinium represented the most abundant reads and dominated the offshore oligotrophic protistan community as they were in the microscopic observations. The dominance of parasitic dinoflagellates in the pyrosequencing dataset, which were overlooked in the morphological observations, indicates more attention should be paid to explore the potential role of this group. Both methods provide coherent clustering of samples. Nutrient levels, salinity and pH were the main factors influencing the distribution of protists. This study demonstrates that different primer pairs used in the pyrosequencing will indicate different protistan community structures. A suitable marker may reveal more comprehensive composition of protists and provide valuable information on environmental drivers.

Detection by real-time PCR and pyrosequencing of the cry1Ab and cry1Ac genes introduced in genetically modified (GM) constructs.

PubMed

Debode, Frederic; Janssen, Eric; Bragard, Claude; Berben, Gilbert

2017-08-01

The presence of genetically modified organisms (GMOs) in food and feed is mainly detected by the use of targets focusing on promoters and terminators. As some genes are frequently used in genetically modified (GM) construction, they also constitute excellent screening elements and their use is increasing. In this paper we propose a new target for the detection of cry1Ab and cry1Ac genes by real-time polymerase chain reaction (PCR) and pyrosequencing. The specificity, sensitivity and robustness of the real-time PCR method were tested following the recommendations of international guidelines and the method met the expected performance criteria. This paper also shows how the robustness testing was assessed. This new cry1Ab/Ac method can provide a positive signal with a larger number of GM events than do the other existing methods using double dye-probes. The method permits the analysis of results with less ambiguity than the SYBRGreen method recommended by the European Reference Laboratory (EURL) GM Food and Feed (GMFF). A pyrosequencing method was also developed to gain additional information thanks to the sequence of the amplicon. This method of sequencing-by-synthesis can determine the sequence between the primers used for PCR. Pyrosequencing showed that the sequences internal to the primers present differences following the GM events considered and three different sequences were observed. The sensitivity of the pyrosequencing was tested on reference flours with a low percentage GM content and different copy numbers. Improvements in the pyrosequencing protocol provided correct sequences with 50 copies of the target. Below this copy number, the quality of the sequence was more random.

The Biosphere.

ERIC Educational Resources Information Center

Cloud, Preston

1983-01-01

Discusses the earth's biosphere, considering how the microbial, animal and plant life (which make up the biosphere) are sustained by the earth's lithosphere, hydrosphere, and atmosphere. Also considers how these three earth features have powerfully shaped the evolution of these organisms. (JN)

Swansong biospheres II: the final signs of life on terrestrial planets near the end of their habitable lifetimes

NASA Astrophysics Data System (ADS)

O'Malley-James, Jack T.; Cockell, Charles S.; Greaves, Jane S.; Raven, John A.

2014-07-01

The biosignatures of life on Earth do not remain static, but change considerably over the planet's habitable lifetime. Earth's future biosphere, much like that of the early Earth, will consist of predominantly unicellular microorganisms due to the increased hostility of environmental conditions caused by the Sun as it enters the late stage of its main sequence evolution. Building on previous work, the productivity of the biosphere is evaluated during different stages of biosphere decline between 1 and 2.8 Gyr from present. A simple atmosphere-biosphere interaction model is used to estimate the atmospheric biomarker gas abundances at each stage and to assess the likelihood of remotely detecting the presence of life in low-productivity, microbial biospheres, putting an upper limit on the lifetime of Earth's remotely detectable biosignatures. Other potential biosignatures such as leaf reflectance and cloud cover are discussed.

Utilizing Pyrosequencing and Quantitative pCR to Characterize Fungal Populations among House Dust Samples

EPA Science Inventory

Molecular techniques are an alternative to culturing and counting methods in quantifying indoor fungal contamination. Pyrosequencing offers the possibility of identifying unexpected indoor fungi. In this study, 50 house dust samples were collected from homes in the Yakima Valley,...

NEW DIRECTIONS: VOCS AND BIOSPHERE-ATMOSPHERE FEEDBACKS

EPA Science Inventory

Shallcross and Monks [New Directions: a Role For Isoprene in Biosphere-Climate-Chemistry Feedbacks, Atmospheric Environment, Vol. 34 (2000) pp. 1659-1660] recently summarized the importance of biogenic isoprene in a biosphere-atmosphere system under constant change. In this art...

Estimating the seasonal carbon source-sink geography of a natural, steady-state terrestrial biosphere

NASA Technical Reports Server (NTRS)

Box, Elgene O.

1988-01-01

The estimation of the seasonal dynamics of biospheric-carbon sources and sinks to be used as an input to global atmospheric CO2 studies and models is discussed. An ecological biosphere model is given and the advantages of the model are examined. Monthly maps of estimated biospheric carbon source and sink regions and estimates of total carbon fluxes are presented for an equilibrium terrestrial biosphere. The results are compared with those from other models. It is suggested that, despite maximum variations of atmospheric CO2 in boreal latitudes, the enormous contributions of tropical wet-dry regions to global atmospheric CO2 seasonality can not be ignored.

Detecting fossil fuel emissions patterns from subcontinental regions using North American in situ CO2 measurements.

PubMed

Shiga, Yoichi P; Michalak, Anna M; Gourdji, Sharon M; Mueller, Kim L; Yadav, Vineet

2014-06-28

The ability to monitor fossil fuel carbon dioxide (FFCO 2 ) emissions from subcontinental regions using atmospheric CO 2 observations remains an important but unrealized goal. Here we explore a necessary but not sufficient component of this goal, namely, the basic question of the detectability of FFCO 2 emissions from subcontinental regions. Detectability is evaluated by examining the degree to which FFCO 2 emissions patterns from specific regions are needed to explain the variability observed in high-frequency atmospheric CO 2 observations. Analyses using a CO 2 monitoring network of 35 continuous measurement towers over North America show that FFCO 2 emissions are difficult to detect during nonwinter months. We find that the compounding effects of the seasonality of atmospheric transport patterns and the biospheric CO 2 flux signal dramatically hamper the detectability of FFCO 2 emissions. Results from several synthetic data case studies highlight the need for advancements in data coverage and transport model accuracy if the goal of atmospheric measurement-based FFCO 2 emissions detection and estimation is to be achieved beyond urban scales. Poor detectability of fossil fuel CO 2 emissions from subcontinental regionsDetectability assessed via attribution of emissions patterns in atmospheric dataLoss in detectability due to transport modeling errors and biospheric signal.

Biosphere 2: The True Story.

ERIC Educational Resources Information Center

O'Keeffe, Michael

1992-01-01

Discusses the history and current developments of the Biosphere 2 Project, a prototype for enclosed self-sustaining structures for space colonization built in the Arizona Desert. Biosphere 2 was created to educate and provide solutions to environmental problems and revenue from research. (MCO)

Effects of climate change on soil moisture over China from 1960-2006

USGS Publications Warehouse

Zhu, Q.; Jiang, H.; Liu, J.

2009-01-01

Soil moisture is an important variable in the climate system and it has sensitive impact on the global climate. Obviously it is one of essential components in the climate change study. The Integrated Biosphere Simulator (IBIS) is used to evaluate the spatial and temporal patterns of soil moisture across China under the climate change conditions for the period 1960-2006. Results show that the model performed better in warm season than in cold season. Mean errors (ME) are within 10% for all the months and root mean squared errors (RMSE) are within 10% except winter season. The model captured the spatial variability higher than 50% in warm seasons. Trend analysis based on the Mann-Kendall method indicated that soil moisture in most area of China is decreased especially in the northern China. The areas with significant increasing trends in soil moisture mainly locate at northwestern China and small areas in southeastern China and eastern Tibet plateau. ?? 2009 IEEE.

Introduction of the hybcell-based compact sequencing technology and comparison to state-of-the-art methodologies for KRAS mutation detection.

PubMed

Zopf, Agnes; Raim, Roman; Danzer, Martin; Niklas, Norbert; Spilka, Rita; Pröll, Johannes; Gabriel, Christian; Nechansky, Andreas; Roucka, Markus

2015-03-01

The detection of KRAS mutations in codons 12 and 13 is critical for anti-EGFR therapy strategies; however, only those methodologies with high sensitivity, specificity, and accuracy as well as the best cost and turnaround balance are suitable for routine daily testing. Here we compared the performance of compact sequencing using the novel hybcell technology with 454 next-generation sequencing (454-NGS), Sanger sequencing, and pyrosequencing, using an evaluation panel of 35 specimens. A total of 32 mutations and 10 wild-type cases were reported using 454-NGS as the reference method. Specificity ranged from 100% for Sanger sequencing to 80% for pyrosequencing. Sanger sequencing and hybcell-based compact sequencing achieved a sensitivity of 96%, whereas pyrosequencing had a sensitivity of 88%. Accuracy was 97% for Sanger sequencing, 85% for pyrosequencing, and 94% for hybcell-based compact sequencing. Quantitative results were obtained for 454-NGS and hybcell-based compact sequencing data, resulting in a significant correlation (r = 0.914). Whereas pyrosequencing and Sanger sequencing were not able to detect multiple mutated cell clones within one tumor specimen, 454-NGS and the hybcell-based compact sequencing detected multiple mutations in two specimens. Our comparison shows that the hybcell-based compact sequencing is a valuable alternative to state-of-the-art methodologies used for detection of clinically relevant point mutations.

DOE Office of Scientific and Technical Information (OSTI.GOV)

None

For nearly 20 years, biosphere reserves have offered a unique framework for building the knowledge, skills, and attitudes required for conservation and sustainable use of ecosystems. The 12 case studies in this volume chronicle many of the cooperative efforts to implement the biosphere reserve concept in the United States. Considered together, these efforts involve more than 20 types of protected areas, and the participation of all levels of government, and many private organizations, academic institutions, citizens groups, and individuals. Biosphere reserves are multi-purpose areas that are nominated by the national committee of the Man and the Biosphere Program (MAB) andmore » designated by the United Nations Educational, Scientific and Cultural Organization (UNESCO) to serve as demonstration areas for cooperation in building harmonious relationships between human activities and the conservation of ecosystems and biological diversity. Each biosphere reserve exemplifies the characteristic ecosystems of one of the worlds biogeographical regions. It is a land or coas%arine area involving human communities as integral components and including resources managed for objectives ranging from complete protection to intensive, yet sustainable development. A biosphere reserve is envisioned as a regional ''landscape for learning'' in which monitoring, research, education, and training are encouraged to support sustainable conservation of natural and managed ecosystems. It is a framework for regional cooperation involving government decisionmakers, scientists, resource managers, private organizations and local people (i.e., the biosphere reserve ''stakeholders''). Finally, each biosphere reserve is part of a global network for sharing information and experience to help address complex problems of conservation and development. The 12 case studies presented in this report represent only a few of the possible evolutions of a biosphere reserve in its efforts to reach out to the local and regional community. As you have read, some have had great success, while others consider their successes almost negligible. All document tremendous effort from many people to improve the communication among landowners, land managers, scientists, and any others interested in the health and well-being of the natural and human environment of the biosphere reserve.« less

Biosphere 2: a prototype project for a permanent and evolving life system for Mars base.

PubMed

Nelson, M; Allen, J P; Dempster, W F

1992-01-01

As part of the ground-based preparation for creating long-term life systems needed for space habitation and settlement, Space Biospheres Ventures (SBV) is undertaking the Biosphere 2 project near Oracle, Arizona. Biosphere 2, currently under construction, is scheduled to commence its operations in 1991 with a two-year closure period with a crew of eight people. Biosphere 2 is a facility which will be essentialy materially-closed to exchange with the outside environment. It is open to information and energy flow. Biosphere 2 is designed to achieve a complex life-support system by the integration of seven areas or "biomes"--rainforest, savannah, desert, marsh, ocean, intensive agriculture and human habitat. Unique bioregenerative technologies, such as soil bed reactors for air purification, aquatic waste processing systems, real-time analytic systems and complex computer monitoring and control systems are being developed for the Biosphere 2 project. Its operation should afford valuable insight into the functioning of complex life systems necessary for long-term habitation in space. It will serve as an experimental ground-based prototype and testbed for the stable, permanent life systems needed for human exploration of Mars.

The First International Biosphere Reserve Congress.

ERIC Educational Resources Information Center

Laird, John

1984-01-01

Presents objectives (and related activities) of a plan designed for international collaboration in conserving key natural areas (biosphere reserves) of the globe. The plan (focusing on such areas as management, conservation, research, monitoring, and environmental education/training) was formulated during the First International Biosphere Reserve…

Global carbon export from the terrestrial biosphere controlled by erosion.

PubMed

Galy, Valier; Peucker-Ehrenbrink, Bernhard; Eglinton, Timothy

2015-05-14

Riverine export of particulate organic carbon (POC) to the ocean affects the atmospheric carbon inventory over a broad range of timescales. On geological timescales, the balance between sequestration of POC from the terrestrial biosphere and oxidation of rock-derived (petrogenic) organic carbon sets the magnitude of the atmospheric carbon and oxygen reservoirs. Over shorter timescales, variations in the rate of exchange between carbon reservoirs, such as soils and marine sediments, also modulate atmospheric carbon dioxide levels. The respective fluxes of biospheric and petrogenic organic carbon are poorly constrained, however, and mechanisms controlling POC export have remained elusive, limiting our ability to predict POC fluxes quantitatively as a result of climatic or tectonic changes. Here we estimate biospheric and petrogenic POC fluxes for a suite of river systems representative of the natural variability in catchment properties. We show that export yields of both biospheric and petrogenic POC are positively related to the yield of suspended sediment, revealing that POC export is mostly controlled by physical erosion. Using a global compilation of gauged suspended sediment flux, we derive separate estimates of global biospheric and petrogenic POC fluxes of 157(+74)(-50) and 43(+61)(-25) megatonnes of carbon per year, respectively. We find that biospheric POC export is primarily controlled by the capacity of rivers to mobilize and transport POC, and is largely insensitive to the magnitude of terrestrial primary production. Globally, physical erosion rates affect the rate of biospheric POC burial in marine sediments more strongly than carbon sequestration through silicate weathering. We conclude that burial of biospheric POC in marine sediments becomes the dominant long-term atmospheric carbon dioxide sink under enhanced physical erosion.

Large scale parallel pyrosequencing technology: PRRSV strain VR-2332 nsp2 deletion mutant stability in swine

USDA-ARS?s Scientific Manuscript database

Genomes from fifteen porcine reproductive and respiratory syndrome virus (PRRSV) isolates were derived simultaneously using 454 pyrosequencing technology. The viral isolates sequenced were from a recent swine study, in which engineered Type 2 prototype PRRSV strain VR-2332 mutants, with 87, 184, 200...

Characterization of chlorinated and chloraminated drinking water microbial communities in a distribution system simulator using pyrosequencing data analysis

EPA Science Inventory

The molecular analysis of drinking water microbial communities has focused primarily on 16S rRNA gene sequence analysis. Since this approach provides limited information on function potential of microbial communities, analysis of whole-metagenome pyrosequencing data was used to...

A morphogenetic survey on ciliate plankton from a mountain lake pinpoints the necessity of lineage-specific barcode markers in microbial ecology.

PubMed

Stoeck, Thorsten; Breiner, Hans-Werner; Filker, Sabine; Ostermaier, Veronika; Kammerlander, Barbara; Sonntag, Bettina

2014-02-01

Analyses of high-throughput environmental sequencing data have become the 'gold-standard' to address fundamental questions of microbial diversity, ecology and biogeography. Findings that emerged from sequencing are, e.g. the discovery of the extensive 'rare microbial biosphere' and its potential function as a seed-bank. Even though applied since several years, results from high-throughput environmental sequencing have hardly been validated. We assessed how well pyrosequenced amplicons [the hypervariable eukaryotic V4 region of the small subunit ribosomal RNA (SSU rRNA) gene] reflected morphotype ciliate plankton. Moreover, we assessed if amplicon sequencing had the potential to detect the annual ciliate plankton stock. In both cases, we identified significant quantitative and qualitative differences. Our study makes evident that taxon abundance distributions inferred from amplicon data are highly biased and do not mirror actual morphotype abundances at all. Potential reasons included cell losses after fixation, cryptic morphotypes, resting stages, insufficient sequence data availability of morphologically described species and the unsatisfying resolution of the V4 SSU rRNA fragment for accurate taxonomic assignments. The latter two underline the necessity of barcoding initiatives for eukaryotic microbes to better and fully exploit environmental amplicon data sets, which then will also allow studying the potential of seed-bank taxa as a buffer for environmental changes. © 2013 The Authors. Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd.

The Study of Socio-Biospheric Problems.

ERIC Educational Resources Information Center

Scott, Andrew M.

Concepts, tools, and a methodology are needed which will permit the analysis of emergent socio-biospheric problems and facilitate their effective management. Many contemporary problems may be characterized as socio-biospheric; for example, pollution of the seas, acid rain, the growth of cities, and an atmosphere loaded with carcinogens. However,…

[The ways in which variations in space and atmospheric factors act upon the biosphere and humans].

PubMed

Chernogor, L F

2010-01-01

The system analysis is validated to be an efficient means for studying the channels through which variations in space and tropospheric weather affect the biosphere (humans). The basics of the system analysis paradigm are presented. The causes of variations in space and tropospheric weather are determined, and the interrelations between them are demonstrated. The ways in which these variations affect the biosphere (humans) are discussed. Aperiodic and quasi-periodic disturbances in the physical fields that influence the biosphere (humans) are intercompared.

Using remote-sensing and the Simple Biosphere model (SiB4) to analyze the seasonality and productivity of the terrestrial biosphere.

NASA Astrophysics Data System (ADS)

Cheeseman, M.; Denning, S.; Baker, I. T.

2017-12-01

Understanding the variability and seasonality of carbon fluxes from the terrestrial biosphere is integral to understanding the mechanisms and drivers of the global carbon cycle. However, there are many regions across the globe where in situ observations are sparse, such as the Amazon rainforest and the African Sahel. The latest version of the Simple-Biosphere model (SiB4) predicts a suite of biophysical variables such as terrestrial carbon flux (GPP), solar induced fluorescence (SIF), fraction of photosynthetically active radiation (FPAR), and leaf area index (LAI). By comparing modeled values to a suite of satellite and in situ observations we produce a robust analysis of the seasonality and productivity of the terrestrial biosphere in a variety of biome types across the globe.

Purification of Marek's disease virus DNA for 454 pyrosequencing using micrococcal nuclease digestion and polyethylene glycol precipitation

USDA-ARS?s Scientific Manuscript database

Marek’s disease virus (MDV-1) is a cell-associated alphaherpesvirus that induces rapid-onset T-cell lymphomas in poultry. The genomes of 6 strains have been sequenced using both Sanger didoxy sequencing and 454 Life Science pyrosequencing. These genomes largely represent cell culture adapted strains...

Pyrosequencing analysis for characterization of bacterial diversity in a soil as affected by integrated livestock-cotton production systems

USDA-ARS?s Scientific Manuscript database

Impacts of integrated livestock-crop production systems compared to specialized systems on soil bacterial diversity have not been well documented. We used a bacterial tag encoded FLX amplicon pyrosequencing (bTEFAP) method to evaluate bacterial diversity of a clay loam soil (Fine, mixed, thermic To...

The water cycle in closed ecological systems: Perspectives from the Biosphere 2 and Laboratory Biosphere systems

NASA Astrophysics Data System (ADS)

Nelson, Mark; Dempster, W. F.; Allen, J. P.

2009-12-01

To achieve sustainable, healthy closed ecological systems requires solutions to challenges of closing the water cycle - recycling wastewater/irrigation water/soil medium leachate and evaporated water and supplying water of required quality as needed for different needs within the facility. Engineering Biosphere 2, the first multi-biome closed ecological system within a total airtight footprint of 12,700 m 2 with a combined volume of 200,000 m 3 with a total water capacity of some 6 × 10 6 L of water was especially challenging because it included human inhabitants, their agricultural and technical systems, as well as five analogue ecosystems ranging from rainforest to desert, freshwater ecologies to saltwater systems like mangrove and mini-ocean coral reef ecosystems. By contrast, the Laboratory Biosphere - a small (40 m 3 volume) soil-based plant growth facility with a footprint of 15 m 2 - is a very simplified system, but with similar challenges re salinity management and provision of water quality suitable for plant growth. In Biosphere 2, water needs included supplying potable water for people and domestic animals, irrigation water for a wide variety of food crops, and recycling and recovering soil nutrients from wastewater. In the wilderness biomes, providing adequately low salinity freshwater terrestrial ecosystems and maintaining appropriate salinity and pH in aquatic/marine ecosystems were challenges. The largest reservoirs in Biosphere 2 were the ocean/marsh with some 4 × 10 6 L, soil with 1 to 2 × 10 6 l, primary storage tank with 0 to 8 × 10 5 L and storage tanks for condensate and soil leachate collection and mixing tanks with a capacity of 1.6 × 10 5 L to supply irrigation for farm and wilderness ecosystems. Other reservoirs were far smaller - humidity in the atmosphere (2 × 10 3 L), streams in the rainforest and savannah, and seasonal pools in the desert were orders of magnitude smaller (8 × 10 4 L). Key technologies included condensation from humidity in the air handlers and from the glass space frame to produce high quality freshwater, wastewater treatment with constructed wetlands and desalination through reverse osmosis and flash evaporation were key to recycling water with appropriate quality throughout the Biosphere 2 facility. Wastewater from all human uses and the domestic animals in Biosphere 2 was treated and recycled through a series of constructed wetlands, which had hydraulic loading of 0.9-1.1 m 3 day -1 (240-290 gal d -1). Plant production in the wetland treatment system produced 1210 kg dry weight of emergent and floating aquatic plant wetland which was used as fodder for the domestic animals while remaining nutrients/water was reused as part of the agricultural irrigation supply. There were pools of water with recycling times of days to weeks and others with far longer cycling times within Biosphere 2. By contrast, the Laboratory Biosphere with a total water reservoir of less than 500 L has far quicker cycling rapidity: for example, atmospheric residence time for water vapor was 5-20 min in the Laboratory Biosphere vs. 1-4 h in Biosphere 2, as compared with 9 days in the Earth's biosphere. Just as in Biosphere 2, humidity in the Laboratory Biosphere amounts to a very small reservoir of water. The amount of water passing through the air in the course of a 12-h operational day is two orders of magnitude greater than the amount stored in the air. Thus, evaporation and condensation collection are vital parts of the recycle system just as in Biosphere 2. The water cycle and sustainable water recycling in closed ecological systems presents problems requiring further research - such as how to control buildup of salinity in materially closed ecosystems and effective ways to retain nutrients in optimal quantity and useable form for plant growth. These issues are common to all closed ecological systems of whatever size, including planet Earth's biosphere and are relevant to a global environment facing increasing water shortages while maintaining water quality for human and ecosystem health. Modular biospheres offer a test bed where technical methods of resolving these problems can be tested for feasibility.

Determination of multiple-clone infection at allelic dimorphism site of Plasmodium vivax merozoite surface protein-1 in the Republic of Korea by pyrosequencing assay.

PubMed

Dinzouna-Boutamba, Sylvatrie-Danne; Lee, Sanghyun; Son, Ui-Han; Yun, Hae Soo; Joo, So-Young; Jeong, Sookwan; Rhee, Man Hee; Kwak, Dongmi; Xuan, Xuenan; Hong, Yeonchul; Chung, Dong-Il; Goo, Youn-Kyoung

2017-12-01

Allelic diversity leading to multiple gene polymorphisms of vivax malaria parasites has been shown to greatly contribute to antigenic variation and drug resistance, increasing the potential for multiple-clone infections within the host. Therefore, to identify multiple-clone infections and the predominant haplotype of Plasmodium vivax in a South Korean population, P. vivax merozoite surface protein-1 (PvMSP-1) was analyzed by pyrosequencing. Pyrosequencing of 156 vivax malaria-infected samples yielded 97 (62.18%) output pyrograms showing two main types of peak patterns of the dimorphic allele for threonine and alanine (T1476A). Most of the samples evaluated (88.66%) carried multiple-clone infections (wild- and mutant-types), whereas 11.34% of the same population carried only the mutant-type (1476A). In addition, each allele showed a high frequency of guanine (G) base substitution at both the first and third positions (86.07% and 81.13%, respectively) of the nucleotide combinations. Pyrosequencing of the PvMSP-1 42-kDa fragment revealed a heterogeneous parasite population, with the mutant-type dominant compared to the wild-type. Understanding the genetic diversity and multiple-clone infection rates may lead to improvements in vivax malaria prevention and strategic control plans. Further studies are needed to improve the efficacy of the pyrosequencing assay with large sample sizes and additional nucleotide positions. Copyright © 2017 Elsevier B.V. All rights reserved.

Governing Portable Conservation and Development Landscapes: Reconsidering Evidence in the Context of the Mbaracayú Biosphere Reserve

ERIC Educational Resources Information Center

Elgert, Laureen

2014-01-01

Conservation-with-development landscapes, such as UNESCO's Man and Biosphere Reserves, differentiate between areas of "nature" and "society". In Paraguay's Mbaracayú Biosphere Reserve, as elsewhere, this model has been used to support governance that focuses on conservation in the "core area" and sustainable…

Selection, management and utilization of biosphere reserves.

Treesearch

Jerry F. Franklin; Stanley L. Krugman

1979-01-01

This publication is directed to the analysis of the selection, management, and utilization of Biosphere Reserves as viewed by scientists from the United States and the Union of Soviet Socialist Republics. Soviet papers focus on types of research and monitoring programs that should be developed on Biosphere Reserves, with emphasis on their use in pollutant monitoring....

WEB-DHM: A distributed biosphere hydrological model developed by coupling a simple biosphere scheme with a hillslope hydrological model

USDA-ARS?s Scientific Manuscript database

The coupling of land surface models and hydrological models potentially improves the land surface representation, benefiting both the streamflow prediction capabilities as well as providing improved estimates of water and energy fluxes into the atmosphere. In this study, the simple biosphere model 2...

Life systems for a lunar base

NASA Technical Reports Server (NTRS)

Nelson, Mark; Hawes, Philip B.; Augustine, Margret

1992-01-01

The Biosphere 2 project is pioneering work on life systems that can serve as a prototype for long-term habitation on the Moon. This project will also facilitate the understanding of the smaller systems that will be needed for initial lunar base life-support functions. In its recommendation for a policy for the next 50 years in space, the National Commission on Space urged, 'To explore and settle the inner Solar System, we must develop biospheres of smaller size, and learn how to build and maintain them' (National Commission on Space, 1986). The Biosphere 2 project, along with its Biospheric Research and Development Center, is a materially closed and informationally and energetically open system capable of supporting a human crew of eight, undertaking work to meet this need. This paper gives an overview of the Space Biospheres Ventures' endeavor and its lunar applications.

Preliminary Feasibility Study of a Hybrid Solar and Modular Pumped Storage Hydro System at Biosphere 2

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lansey, Kevin; Hortsman, Chris

2016-10-01

In this study, the preliminary feasibility of a hybrid solar and modular pumped storage system designed for high energy independence at Biosphere 2 is assessed. The system consists of an array of solar PV panels that generate electricity during the day to power both Biosphere 2 and a pump that sends water through a pipe to a tank at a high elevation. When solar power is not available, the water is released back down the pipe towards a tank at a lower elevation, where it passes through a hydraulic water turbine to generate hydroelectricity to power Biosphere 2. The hybridmore » system is sized to generate and store enough energy to enable Biosphere 2 to operate without a grid interconnection on an average day.« less

Biosphere as a complex life-support system (LSS) for human civilization

NASA Astrophysics Data System (ADS)

Pechurkin, Nickolay

As a continuously growing link of the Biosphere, we should keep in mind that biotic cycles induced by flows of a solar energy are the source of Biosphere and ecosystems functioning. Our pressure on the Biosphere which is connected with biotic cycle’s alterations and damages is menacingly growing. There are innumerable examples of atmosphere, water and soil pollution. We have contaminated even Earth-Space orbits with different uncontrolled debris. Ecological Footprint (EF) is a proper quantitative measure of anthropogenic impact on the Biosphere and ecosystems functioning. The comparative dynamics of the United Nations’ Human Development Index (HDI) and Ecological Footprint (EF) is discussed in the paper. The main call of sustainable development of mankind: all humans can have opportunity to fulfill their lives without degrading the Biosphere. To support sustainability, we should make an effort to develop each nation and the mankind as a whole with a high HDI and with a low ecological footprint. It means: to have high level of HDI at low level of EF. But current tendency of economical and social development shows: the higher HDI, the bigger EF. EF of mankind is rising threateningly. Now actual pressure of the human civilization of our planet (2014) upon 60 % exceeds its potential possibilities (biological capacity, measured as the area of "global" green hectares). It means that now we require more than 1.5 planets of the Earth’s type for sustainable development. It leads to ecological incident in the scale of Biosphere. Our Biosphere is the large, multilevel, hierarchically organized system, and our civilization is only a part of it. This part is not central; it can disappear for ever, if we do not cope to be included in the Biosphere as a great complex system. An example of Krasnoyarsk region as a representative region with high level of industry and technological energy production is considered in the paper. This work was supported by the Russian Foundation for Basic Research, project number 13-06-00060.

454 pyrosequencing project identifying expressed genes from the horn fly, Haematobia irritans

USDA-ARS?s Scientific Manuscript database

We used an EST approach to initiate a study of the genome of the horn fly, Haematobia irritans and have used 454 pyrosequencing techniques to sequence 73,512, 100,603, 71,550, and 85,769 expressed genes from the egg, first instar larvae, adult male, and adult female lifestages of the horn fly. cD...

Pyrosequencing of the northern red oak (Quercus rubra L.) chloroplast genome reveals high quality polymorphisms for population management

Treesearch

Lisa W. Alexander; Keith E. Woeste

2014-01-01

Given the low intraspecific chloroplast diversity detected in northern red oak (Quercus rubra L.), more powerful genetic tools are necessary to accurately characterize Q. rubra chloroplast diversity and structure. We report the sequencing, assembly, and annotation of the chloroplast genome of northern red oak via pyrosequencing and...

Assessment of bacterial contamination of lipstick using pyrosequencing.

PubMed

Lee, So Y; Lee, Si Y

As soon as they are exposed to the environment, cosmetics become contaminated with microorganisms, and this contamination accumulates with increased use. In this study, we employed pyrosequencing to investigate the diversity of bacteria found on lipstick. Bacterial DNA was extracted from 20 lipstick samples and mixed in equal ratios for pyrosequencing analysis. As a result, 105 bacterial genera were detected, four of which ( Leifsonia , Methylobacterium , Streptococcus , and Haemophilus ) were predominant in 92% of the 19,863 total sequence reads. Potentially pathogenic genera such as Staphylococcus , Pseudomonas , Escherichia , Salmonella , Corynebacterium , Mycobacterium , and Neisseria accounted for 27.6% of the 105 genera. The most commonly identified oral bacteria belonged to the Streptococcus genus, although other oral genera such as Actinomyces , Fusobacterium , Porphyromonas , and Lactobacillus were also detected.

Comparative Analysis of Daytime Fire Detection Algorithms, Using AVHRR Data for the 1995 Fire Season in Canda: Perspective for MODIS

NASA Technical Reports Server (NTRS)

Ichoku, Charles; Kaufman, Y. J.; Fraser, R. H.; Jin, J.-Z.; Park, W. M.; Lau, William K. M. (Technical Monitor)

2001-01-01

Two fixed-threshold Canada Centre for Remote Sensing and European Space Agency (CCRS and ESA) and three contextual GIGLIO, International Geosphere and Biosphere Project, and Moderate Resolution Imaging Spectroradiometer (GIGLIO, IGBP, and MODIS) algorithms were used for fire detection with Advanced Very High Resolution Radiometer (AVHRR) data acquired over Canada during the 1995 fire season. The CCRS algorithm was developed for the boreal ecosystem, while the other four are for global application. The MODIS algorithm, although developed specifically for use with the MODIS sensor data, was applied to AVHRR in this study for comparative purposes. Fire detection accuracy assessment for the algorithms was based on comparisons with available 1995 burned area ground survey maps covering five Canadian provinces. Overall accuracy estimations in terms of omission (CCRS=46%, ESA=81%, GIGLIO=75%, IGBP=51%, MODIS=81%) and commission (CCRS=0.35%, ESA=0.08%, GIGLIO=0.56%, IGBP=0.75%, MODIS=0.08%) errors over forested areas revealed large differences in performance between the algorithms, with no relevance to type (fixed-threshold or contextual). CCRS performed best in detecting real forest fires, with the least omission error, while ESA and MODIS produced the highest omission error, probably because of their relatively high threshold values designed for global application. The commission error values appear small because the area of pixels falsely identified by each algorithm was expressed as a ratio of the vast unburned forest area. More detailed study shows that most commission errors in all the algorithms were incurred in nonforest agricultural areas, especially on days with very high surface temperatures. The advantage of the high thresholds in ESA and MODIS was that they incurred the least commission errors.

Spanish methodological approach for biosphere assessment of radioactive waste disposal.

PubMed

Agüero, A; Pinedo, P; Cancio, D; Simón, I; Moraleda, M; Pérez-Sánchez, D; Trueba, C

2007-10-01

The development of radioactive waste disposal facilities requires implementation of measures that will afford protection of human health and the environment over a specific temporal frame that depends on the characteristics of the wastes. The repository design is based on a multi-barrier system: (i) the near-field or engineered barrier, (ii) far-field or geological barrier and (iii) the biosphere system. Here, the focus is on the analysis of this last system, the biosphere. A description is provided of conceptual developments, methodological aspects and software tools used to develop the Biosphere Assessment Methodology in the context of high-level waste (HLW) disposal facilities in Spain. This methodology is based on the BIOMASS "Reference Biospheres Methodology" and provides a logical and systematic approach with supplementary documentation that helps to support the decisions necessary for model development. It follows a five-stage approach, such that a coherent biosphere system description and the corresponding conceptual, mathematical and numerical models can be built. A discussion on the improvements implemented through application of the methodology to case studies in international and national projects is included. Some facets of this methodological approach still require further consideration, principally an enhanced integration of climatology, geography and ecology into models considering evolution of the environment, some aspects of the interface between the geosphere and biosphere, and an accurate quantification of environmental change processes and rates.

Man and the Biosphere: Ground Truthing Coral Reefs for the St. John Island Biosphere Reserve.

ERIC Educational Resources Information Center

Brody, Michael J.; And Others

Research on the coral species composition of St. John's reefs in the Virgin Islands was conducted through the School for Field Studies (SFS) Coral Reef Ecology course (winter 1984). A cooperative study program based on the United Nations Educational, Scientific, and Cultural Organization's (Unesco) program, Man and the Biosphere, was undertaken by…

Bayesian inversions of a dynamic vegetation model at four European grassland sites

NASA Astrophysics Data System (ADS)

Minet, J.; Laloy, E.; Tychon, B.; Francois, L.

2015-05-01

Eddy covariance data from four European grassland sites are used to probabilistically invert the CARAIB (CARbon Assimilation In the Biosphere) dynamic vegetation model (DVM) with 10 unknown parameters, using the DREAM(ZS) (DiffeRential Evolution Adaptive Metropolis) Markov chain Monte Carlo (MCMC) sampler. We focus on comparing model inversions, considering both homoscedastic and heteroscedastic eddy covariance residual errors, with variances either fixed a priori or jointly inferred together with the model parameters. Agreements between measured and simulated data during calibration are comparable with previous studies, with root mean square errors (RMSEs) of simulated daily gross primary productivity (GPP), ecosystem respiration (RECO) and evapotranspiration (ET) ranging from 1.73 to 2.19, 1.04 to 1.56 g C m-2 day-1 and 0.50 to 1.28 mm day-1, respectively. For the calibration period, using a homoscedastic eddy covariance residual error model resulted in a better agreement between measured and modelled data than using a heteroscedastic residual error model. However, a model validation experiment showed that CARAIB models calibrated considering heteroscedastic residual errors perform better. Posterior parameter distributions derived from using a heteroscedastic model of the residuals thus appear to be more robust. This is the case even though the classical linear heteroscedastic error model assumed herein did not fully remove heteroscedasticity of the GPP residuals. Despite the fact that the calibrated model is generally capable of fitting the data within measurement errors, systematic bias in the model simulations are observed. These are likely due to model inadequacies such as shortcomings in the photosynthesis modelling. Besides the residual error treatment, differences between model parameter posterior distributions among the four grassland sites are also investigated. It is shown that the marginal distributions of the specific leaf area and characteristic mortality time parameters can be explained by site-specific ecophysiological characteristics.

Hybrid inversions of CO2 fluxes at regional scale applied to network design

NASA Astrophysics Data System (ADS)

Kountouris, Panagiotis; Gerbig, Christoph; -Thomas Koch, Frank

2013-04-01

Long term observations of atmospheric greenhouse gas measuring stations, located at representative regions over the continent, improve our understanding of greenhouse gas sources and sinks. These mixing ratio measurements can be linked to surface fluxes by atmospheric transport inversions. Within the upcoming years new stations are to be deployed, which requires decision making tools with respect to the location and the density of the network. We are developing a method to assess potential greenhouse gas observing networks in terms of their ability to recover specific target quantities. As target quantities we use CO2 fluxes aggregated to specific spatial and temporal scales. We introduce a high resolution inverse modeling framework, which attempts to combine advantages from pixel based inversions with those of a carbon cycle data assimilation system (CCDAS). The hybrid inversion system consists of the Lagrangian transport model STILT, the diagnostic biosphere model VPRM and a Bayesian inversion scheme. We aim to retrieve the spatiotemporal distribution of net ecosystem exchange (NEE) at a high spatial resolution (10 km x 10 km) by inverting for spatially and temporally varying scaling factors for gross ecosystem exchange (GEE) and respiration (R) rather than solving for the fluxes themselves. Thus the state space includes parameters for controlling photosynthesis and respiration, but unlike in a CCDAS it allows for spatial and temporal variations, which can be expressed as NEE(x,y,t) = λG(x,y,t) GEE(x,y,t) + λR(x,y,t) R(x,y,t) . We apply spatially and temporally correlated uncertainties by using error covariance matrices with non-zero off-diagonal elements. Synthetic experiments will test our system and select the optimal a priori error covariance by using different spatial and temporal correlation lengths on the error statistics of the a priori covariance and comparing the optimized fluxes against the 'known truth'. As 'known truth' we use independent fluxes generated from a different biosphere model (BIOME-BGC). Initially we perform single-station inversions for Ochsenkopf tall tower located in Germany. Further expansion of the inversion framework to multiple stations and its application to network design will address the questions of how well a set of network stations can constrain a given target quantity, and whether there are objective criteria to select an optimal configuration for new stations that maximizes the uncertainty reduction.

Using the kingfisher (Alcedo atthis) as a bioindicator of PCBs and PBDEs in the dinghushan biosphere reserve, China.

PubMed

Mo, Ling; Wu, Jiang-Ping; Luo, Xiao-Jun; Li, Ke-Lin; Peng, Ying; Feng, An-Hong; Zhang, Qiang; Zou, Fa-Sheng; Mai, Bi-Xian

2013-07-01

The Dinghushan Biosphere Reserve is a nature reserve and a site for the study of tropical and subtropical forest ecosystems. Rapid industrialization and intensive electronic waste-recycling activities around the biosphere reserve have resulted in elevated levels of industrial organic contaminants in the local environment that may cause adverse effects on wildlife that inhabits this area. In the present study, polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and 2 alternative brominated flame retardants (BFRs)-decabromodiphenyl ethane (DBDPE) and 1,2-bis(2,4,6-tribromophenoxy) ethane (BTBPE)-were investigated in the biosphere reserve and a reference site by using the kingfisher (Alcedo atthis) as a bioindicator. Residue concentrations in kingfishers from the Dinghushan Biosphere Reserve ranged from 490 ng/g to 3000 ng/g, 51 ng/g to 420 ng/g, 0.44 ng/g to 90 ng/g, and 0.04 ng/g to 0.87 ng/g lipid weight for ∑PCBs, ∑PBDEs, DBDPE, and BTBPE, respectively. With the exception of the BTBPE, these levels were 2 to 5 times higher than those detected in kingfishers from the reference site. The contaminant patterns from the biosphere reserve were also different, with larger PCB contributions in comparison with the reference site. The estimated predator-prey biomagnification factors (BMFs) showed that most of the PCB and PBDE congeners and BTBPE were biomagnified in kingfishers from the biosphere reserve. The calculated toxic equivalent quantity (TEQ) concentrations of major coplanar PCB congeners in kingfishers from the biosphere reserve ranged from 18 pg/g to 66 pg/g wet weight, with some of these TEQ concentrations reaching or exceeding the levels known to impair bird reproduction and survival. Copyright © 2013 SETAC.

On the Ability of Space-Based Passive and Active Remote Sensing Observations of CO2 to Detect Flux Perturbations to the Carbon Cycle

NASA Astrophysics Data System (ADS)

Crowell, Sean M. R.; Randolph Kawa, S.; Browell, Edward V.; Hammerling, Dorit M.; Moore, Berrien; Schaefer, Kevin; Doney, Scott C.

2018-01-01

Space-borne observations of CO2 are vital to gaining understanding of the carbon cycle in regions of the world that are difficult to measure directly, such as the tropical terrestrial biosphere, the high northern and southern latitudes, and in developing nations such as China. Measurements from passive instruments such as GOSAT and OCO-2, however, are constrained by solar zenith angle limitations as well as sensitivity to the presence of clouds and aerosols. Active measurements such as those in development for the Active Sensing of CO2 Emissions over Nights, Days and Seasons (ASCENDS) mission show strong potential for making measurements in the high-latitude winter and in cloudy regions. In this work we examine the enhanced flux constraint provided by the improved coverage from an active measurement such as ASCENDS. The simulation studies presented here show that with sufficient precision, ASCENDS will detect permafrost thaw and fossil fuel emissions shifts at annual and seasonal time scales, even in the presence of transport errors, representativeness errors, and biogenic flux errors. While OCO-2 can detect some of these perturbations at the annual scale, the seasonal sampling provided by ASCENDS provides the stronger constraint.

The biosphere: Problems and solutions; Proceedings of the Miami International Symposium on the Biosphere, Miami Beach, FL, April 23, 24, 1984

NASA Astrophysics Data System (ADS)

Veziroglu, T. N.

The objective of the Miami International Symposium on the Biosphere was to provide a forum for the presentation of the latest research findings on the environmental effects of human activities. The topics discussed are related to biosphere reserves, environmental aspects of hydrocarbon fuels, radioactivity and nuclear waste, land management, acid rains, water quality, water resources, coastal resources management, the pollution of rivers, industrial waste, economic development and the environment, health hazards and solutions, endangered species, environmentally compatible systems, space pollution, and global considerations. Attention is given to questions regarding global security and sustainable development, environethics as a global strategy for environmental quality, a gestalt approach to the environment, potential indicators for monitoring biosphere reserves, a review of regional impacts associated with the development of U.S. synthetic fuel resources, water resources in the Soviet Union, and pollution-free pesticides.

N-ras Mutation Detection by Pyrosequencing in Adult Patients with Acute Myeloid Leukemia at a Single Institution

PubMed Central

Jeong, Ji Hun; Park, Soon Ho; Park, Mi Jung; Kim, Moon Jin; Kim, Kyung Hee; Park, Pil Whan; Seo, Yiel Hea; Lee, Jae Hoon; Park, Jinny; Hong, Junshik

2013-01-01

Background N-ras mutations are one of the most commonly detected abnormalities of myeloid origin. N-ras mutations result in a constitutively active N-ras protein that induces uncontrolled cell proliferation and inhibits apoptosis. We analyzed N-ras mutations in adult patients with AML at a particular institution and compared pyrosequencing analysis with a direct sequencing method for the detection of N-ras mutations. Methods We analyzed 90 bone marrow samples from 83 AML patients. We detected N-ras mutations in codons 12, 13, and 61 using the pyrosequencing method and subsequently confirmed all data by direct sequencing. Using these methods, we screened the N-ras mutation quantitatively and determined the incidence and characteristic of N-ras mutation. Results The incidence of N-ras mutation was 7.2% in adult AML patients. The patients with N-ras mutations showed significant higher hemoglobin levels (P=0.022) and an increased incidence of FLT3 mutations (P=0.003). We observed 3 cases with N-ras mutations in codon 12 (3.6%), 2 cases in codon 13 (2.4%), and 1 case in codon 61 (1.2%). All the mutations disappeared during chemotherapy. Conclusions There is a low incidence (7.2%) of N-ras mutations in AML patients compared with other populations. Similar data is obtained by both pyrosequencing and direct sequencing. This study showed the correlation between the N-ras mutation and the therapeutic response. However, pyrosequencing provides quantitative data and is useful for monitoring therapeutic responses. PMID:23667841

454-Pyrosequencing survey of microbiota in adult Spotted Wing Drosophila (SWD) corroborates a core microbiome and additional symbiotic and entomopathogenic bacterial associates

USDA-ARS?s Scientific Manuscript database

Complete surveys of insect endosymbionts including species of economic importance have until recently been hampered by a lack of high-throughput genetic assays. We used 454-pyrosequencing of the 16S rRNA gene amplicon of adult spotted wing Drosophila (SWD) Drosophila suzukii (Matsumura) from souther...

Development of colonic microflora as assessed by pyrosequencing in dairy calves fed waste milk

USDA-ARS?s Scientific Manuscript database

The objective of the current study was to examine the effect of pasteurization of waste milk used to feed dairy calves on the bacterial diversity of their lower gut. Using 16S rDNA bacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP), fecal samples from dairy calves aging from 1 week to 6 mon...

Microbial analysis in primary and persistent endodontic infections by using pyrosequencing.

PubMed

Hong, Bo-Young; Lee, Tae-Kwon; Lim, Sang-Min; Chang, Seok Woo; Park, Joonhong; Han, Seung Hyun; Zhu, Qiang; Safavi, Kamran E; Fouad, Ashraf F; Kum, Kee Yeon

2013-09-01

The aim of this study was to investigate the bacterial community profile of intracanal microbiota in primary and persistent endodontic infections associated with asymptomatic chronic apical periodontitis by using GS-FLX Titanium pyrosequencing. The null hypothesis was that there is no difference in diversity of overall bacterial community profiles between primary and persistent infections. Pyrosequencing analysis from 10 untreated and 8 root-filled samples was conducted. Analysis from 18 samples yielded total of 124,767 16S rRNA gene sequences (with a mean of 6932 reads per sample) that were taxonomically assigned into 803 operational taxonomic units (3% distinction), 148 genera, and 10 phyla including unclassified. Bacteroidetes was the most abundant phylum in both primary and persistent infections. There were no significant differences in bacterial diversity between the 2 infection groups (P > .05). The bacterial community profile that was based on dendrogram showed that bacterial population in both infections was not significantly different in their structure and composition (P > .05). The present pyrosequencing study demonstrates that persistent infections have as diverse bacterial community as primary infections. Copyright © 2013 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Assessment of the microbial diversity of Brazilian kefir grains by PCR-DGGE and pyrosequencing analysis.

PubMed

Leite, A M O; Mayo, B; Rachid, C T C C; Peixoto, R S; Silva, J T; Paschoalin, V M F; Delgado, S

2012-09-01

The microbial diversity and community structure of three different kefir grains from different parts of Brazil were examined via the combination of two culture-independent methods: PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and pyrosequencing. PCR-DGGE showed Lactobacillus kefiranofaciens and Lactobacillus kefiri to be the major bacterial populations in all three grains. The yeast community was dominated by Saccharomyces cerevisiae. Pyrosequencing produced a total of 14,314 partial 16S rDNA sequence reads from the three grains. Sequence analysis grouped the reads into three phyla, of which Firmicutes was dominant. Members of the genus Lactobacillus were the most abundant operational taxonomic units (OTUs) in all samples, accounting for up to 96% of the sequences. OTUs belonging to other lactic and acetic acid bacteria genera, such as Lactococcus, Leuconostoc, Streptococcus and Acetobacter, were also identified at low levels. Two of the grains showed identical DGGE profiles and a similar number of OTUs, while the third sample showed the highest diversity by both techniques. Pyrosequencing allowed the identification of bacteria that were present in small numbers and rarely associated with the microbial community of this complex ecosystem. Copyright © 2012 Elsevier Ltd. All rights reserved.

Rapid phylogenetic dissection of prokaryotic community structure in tidal flat using pyrosequencing.

PubMed

Kim, Bong-Soo; Kim, Byung Kwon; Lee, Jae-Hak; Kim, Myungjin; Lim, Young Woon; Chun, Jongsik

2008-08-01

Dissection of prokaryotic community structure is prerequisite to understand their ecological roles. Various methods are available for such a purpose which amplification and sequencing of 16S rRNA genes gained its popularity. However, conventional methods based on Sanger sequencing technique require cloning process prior to sequencing, and are expensive and labor-intensive. We investigated prokaryotic community structure in tidal flat sediments, Korea, using pyrosequencing and a subsequent automated bioinformatic pipeline for the rapid and accurate taxonomic assignment of each amplicon. The combination of pyrosequencing and bioinformatic analysis showed that bacterial and archaeal communities were more diverse than previously reported in clone library studies. Pyrosequencing analysis revealed 21 bacterial divisions and 37 candidate divisions. Proteobacteria was the most abundant division in the bacterial community, of which Gamma-and Delta-Proteobacteria were the most abundant. Similarly, 4 archaeal divisions were found in tidal flat sediments. Euryarchaeota was the most abundant division in the archaeal sequences, which were further divided into 8 classes and 11 unclassified euryarchaeota groups. The system developed here provides a simple, in-depth and automated way of dissecting a prokaryotic community structure without extensive pretreatment such as cloning.

Clinical Neuropathology practice news 1-2014: Pyrosequencing meets clinical and analytical performance criteria for routine testing of MGMT promoter methylation status in glioblastoma

PubMed Central

Preusser, Matthias; Berghoff, Anna S.; Manzl, Claudia; Filipits, Martin; Weinhäusel, Andreas; Pulverer, Walter; Dieckmann, Karin; Widhalm, Georg; Wöhrer, Adelheid; Knosp, Engelbert; Marosi, Christine; Hainfellner, Johannes A.

2014-01-01

Testing of the MGMT promoter methylation status in glioblastoma is relevant for clinical decision making and research applications. Two recent and independent phase III therapy trials confirmed a prognostic and predictive value of the MGMT promoter methylation status in elderly glioblastoma patients. Several methods for MGMT promoter methylation testing have been proposed, but seem to be of limited test reliability. Therefore, and also due to feasibility reasons, translation of MGMT methylation testing into routine use has been protracted so far. Pyrosequencing after prior DNA bisulfite modification has emerged as a reliable, accurate, fast and easy-to-use method for MGMT promoter methylation testing in tumor tissues (including formalin-fixed and paraffin-embedded samples). We performed an intra- and inter-laboratory ring trial which demonstrates a high analytical performance of this technique. Thus, pyrosequencing-based assessment of MGMT promoter methylation status in glioblastoma meets the criteria of high analytical test performance and can be recommended for clinical application, provided that strict quality control is performed. Our article summarizes clinical indications, practical instructions and open issues for MGMT promoter methylation testing in glioblastoma using pyrosequencing. PMID:24359605

Development of a distributed biosphere hydrological model and its evaluation with the Southern Great Plains Experiments (SGP97 and SGP99)

USDA-ARS?s Scientific Manuscript database

A distributed biosphere hydrological model, the so called water and energy budget-based distributed hydrological model (WEB-DHM), has been developed by fully coupling a biosphere scheme (SiB2) with a geomorphology-based hydrological model (GBHM). SiB2 describes the transfer of turbulent fluxes (ener...

The Biosphere: A Decadal Vision

NASA Technical Reports Server (NTRS)

Peterson, David L.; Curran, Paul J.; Mlynzcak, Marty; Miller, Richard

2003-01-01

This paper focuses on biosphere-climate interactions including the influences of human activities. Recognizing this is only one aspect of biospheric processes, this places an emphasis of those biogeochemical processes that have a profound effect on numerous other aspects of the biosphere and the services it provides, services which are critical to sustaining life on Earth. And, the paper will focus on the various scientific aspects of assessing the availability of fresh water, including its sensitivity to climate variance and land use changes. Finally, this paper hopes to emphasize the potential role that greatly expanded space observations and interactive modeling can play in developing our understanding of Earth and its the living systems.

Biosphere 2 test module experimentation program

NASA Technical Reports Server (NTRS)

Alling, Abigail; Leigh, Linda S.; Maccallum, Taber; Alvarez-Romo, Norberto

1990-01-01

The Biosphere 2 Test Module is a facility which has the capability to do either short or long term closures: five month closures with plants were conducted. Also conducted were investigations of specific problems, such as trace gas purification by bioregenerative systems by in-putting a fixed concentration of a gas and observing its uptake over time. In other Test Module experiments, the concentration of one gas was changed to observe what effects this has on other gases present or on the system. The science of biospherics which encompasses the study of closed biological systems provides an opening into the future in space as well as in the Earth's biosphere.

Rarity in aquatic microbes: placing protists on the map.

PubMed

Logares, Ramiro; Mangot, Jean-François; Massana, Ramon

2015-12-01

Most microbial richness at any given time tends to be represented by low-abundance (rare) taxa, which are collectively referred to as the "rare biosphere". Here we review works on the rare biosphere using high-throughput sequencing (HTS), with a particular focus on unicellular eukaryotes or protists. Evidence thus far indicates that the rare biosphere encompasses dormant as well as metabolically active microbes that could potentially play key roles in ecosystem functioning. Rare microbes appear to have biogeography, and sometimes the observed patterns can be similar to what is observed among abundant taxa, suggesting similar community-structuring mechanisms. There is limited evidence indicating that the rare biosphere contains taxa that are phylogenetically distantly related to abundant counterparts; therefore, the rare biosphere may act as a reservoir of deep-branching phylogenetic diversity. The potential role of the rare biosphere as a bank of redundant functions that can help to maintain continuous ecosystem function following oscillations in taxonomic abundances is hypothesized as its main ecological role. Future studies focusing on rare microbes are crucial for advancing our knowledge of microbial ecology and evolution and unveiling their links with ecosystem function. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Opportunities and challenges in studies of deep life (Invited)

NASA Astrophysics Data System (ADS)

Edwards, K. J.

2010-12-01

Over the past two decades, there has been an increasing awareness within the geological, microbiological, and oceanographic communities of the potentially vast microbial biosphere that is harbored beneath the surface of the Earth. With this awareness has come a mounting effort to study this potential biome - to better quantify biomass abundance, activity, and biogeochemical activity. In the Earth system, the largest deep subsurface biome is also the least accessible - the deep ocean subsurface biosphere. The oceanic deep biosphere also has greatest potential for influencing global scale biogeochemical processes -the carbon and energy cycles for example, and other elemental cycles. To address these topics and mount interdisciplinary efforts to study the deep subsurface marine biosphere, we have recently formed a center in support integrative, collaborative investigations. The national science foundation Center for Dark Biosphere Investigations (C-DEBI), has been initiated for the explicit purpose of resolving the extent, function, dynamics and implications of the subseafloor biosphere. This talk will discuss C-DEBI science, with focus on some of the opportunities and challenges in the study of deep life in the ocean, and the role that C-DEBI will play in meeting them

Analysis of genetically modified organisms by pyrosequencing on a portable photodiode-based bioluminescence sequencer.

PubMed

Song, Qinxin; Wei, Guijiang; Zhou, Guohua

2014-07-01

A portable bioluminescence analyser for detecting the DNA sequence of genetically modified organisms (GMOs) was developed by using a photodiode (PD) array. Pyrosequencing on eight genes (zSSIIb, Bt11 and Bt176 gene of genetically modified maize; Lectin, 35S-CTP4, CP4EPSPS, CaMV35S promoter and NOS terminator of the genetically modified Roundup ready soya) was successfully detected with this instrument. The corresponding limit of detection (LOD) was 0.01% with 35 PCR cycles. The maize and soya available from three different provenances in China were detected. The results indicate that pyrosequencing using the small size of the detector is a simple, inexpensive, and reliable way in a farm/field test of GMO analysis. Copyright © 2014 Elsevier Ltd. All rights reserved.

Response of the rare biosphere to environmental stressors in a highly diverse ecosystem (Zodletone spring, OK, USA).

PubMed

Coveley, Suzanne; Elshahed, Mostafa S; Youssef, Noha H

2015-01-01

Within highly diverse ecosystems, the majority of bacterial taxa are present in low abundance as members of the rare biosphere. The rationale for the occurrence and maintenance of the rare biosphere, and the putative ecological role(s) and dynamics of its members within a specific ecosystem is currently debated. We hypothesized that in highly diverse ecosystems, a fraction of the rare biosphere acts as a backup system that readily responds to environmental disturbances. We tested this hypothesis by subjecting sediments from Zodletone spring, a sulfide- and sulfur-rich spring in Southwestern OK, to incremental levels of salinity (1, 2, 3, 4, and 10% NaCl), or temperature (28°, 30°, 32°, and 70 °C), and traced the trajectories of rare members of the community in response to these manipulations using 16S rRNA gene analysis. Our results indicate that multiple rare bacterial taxa are promoted from rare to abundant members of the community following such manipulations and that, in general, the magnitude of such recruitment is directly proportional to the severity of the applied manipulation. Rare members that are phylogenetically distinct from abundant taxa in the original sample (unique rare biosphere) played a more important role in the microbial community response to environmental disturbances, compared to rare members that are phylogenetically similar to abundant taxa in the original sample (non-unique rare biosphere). The results emphasize the dynamic nature of the rare biosphere, and highlight its complexity and non-monolithic nature.

Analysis of RET promoter CpG island methylation using methylation-specific PCR (MSP), pyrosequencing, and methylation-sensitive high-resolution melting (MS-HRM): impact on stage II colon cancer patient outcome.

PubMed

Draht, Muriel X G; Smits, Kim M; Jooste, Valérie; Tournier, Benjamin; Vervoort, Martijn; Ramaekers, Chantal; Chapusot, Caroline; Weijenberg, Matty P; van Engeland, Manon; Melotte, Veerle

2016-01-01

Already since the 1990s, promoter CpG island methylation markers have been considered promising diagnostic, prognostic, and predictive cancer biomarkers. However, so far, only a limited number of DNA methylation markers have been introduced into clinical practice. One reason why the vast majority of methylation markers do not translate into clinical applications is lack of independent validation of methylation markers, often caused by differences in methylation analysis techniques. We recently described RET promoter CpG island methylation as a potential prognostic marker in stage II colorectal cancer (CRC) patients of two independent series. In the current study, we analyzed the RET promoter CpG island methylation of 241 stage II colon cancer patients by direct methylation-specific PCR (MSP), nested-MSP, pyrosequencing, and methylation-sensitive high-resolution melting (MS-HRM). All primers were designed as close as possible to the same genomic region. In order to investigate the effect of different DNA methylation assays on patient outcome, we assessed the clinical sensitivity and specificity as well as the association of RET methylation with overall survival for three and five years of follow-up. Using direct-MSP and nested-MSP, 12.0 % (25/209) and 29.6 % (71/240) of the patients showed RET promoter CpG island methylation. Methylation frequencies detected by pyrosequencing were related to the threshold for positivity that defined RET methylation. Methylation frequencies obtained by pyrosequencing (threshold for positivity at 20 %) and MS-HRM were 13.3 % (32/240) and 13.8 % (33/239), respectively. The pyrosequencing threshold for positivity of 20 % showed the best correlation with MS-HRM and direct-MSP results. Nested-MSP detected RET promoter CpG island methylation in deceased patients with a higher sensitivity (33.1 %) compared to direct-MSP (10.7 %), pyrosequencing (14.4 %), and MS-HRM (15.4 %). While RET methylation frequencies detected by nested-MSP, pyrosequencing, and MS-HRM varied, the prognostic effect seemed similar (HR 1.74, 95 % CI 0.97-3.15; HR 1.85, 95 % CI 0.93-3.86; HR 1.83, 95 % CI 0.92-3.65, respectively). Our results show that upon optimizing and aligning four RET methylation assays with regard to primer location and sensitivity, differences in methylation frequencies and clinical sensitivities are observed; however, the effect on the marker's prognostic outcome is minimal.

Access: A Directory of Contacts, Environmental Data Bases, and Scientific Infrastructure on 175 Biosphere Reserves in 32 Countries.

ERIC Educational Resources Information Center

Department of State, Washington, DC. Bureau of Oceans and International Environmental and Scientific Affairs.

Following the EuroMAB meeting in Strasbourg, France (September 1991) and on an initiative of the Man and the Biosphere National Committee of the United States, a decision was made to create a research network from information available in biosphere reserves in 30 European countries, Canada and the United States. This Directory of EuroMAB Biosphere…

Post-closure biosphere assessment modelling: comparison of complex and more stylised approaches.

PubMed

Walke, Russell C; Kirchner, Gerald; Xu, Shulan; Dverstorp, Björn

2015-10-01

Geological disposal facilities are the preferred option for high-level radioactive waste, due to their potential to provide isolation from the surface environment (biosphere) on very long timescales. Assessments need to strike a balance between stylised models and more complex approaches that draw more extensively on site-specific information. This paper explores the relative merits of complex versus more stylised biosphere models in the context of a site-specific assessment. The more complex biosphere modelling approach was developed by the Swedish Nuclear Fuel and Waste Management Co (SKB) for the Formark candidate site for a spent nuclear fuel repository in Sweden. SKB's approach is built on a landscape development model, whereby radionuclide releases to distinct hydrological basins/sub-catchments (termed 'objects') are represented as they evolve through land rise and climate change. Each of seventeen of these objects is represented with more than 80 site specific parameters, with about 22 that are time-dependent and result in over 5000 input values per object. The more stylised biosphere models developed for this study represent releases to individual ecosystems without environmental change and include the most plausible transport processes. In the context of regulatory review of the landscape modelling approach adopted in the SR-Site assessment in Sweden, the more stylised representation has helped to build understanding in the more complex modelling approaches by providing bounding results, checking the reasonableness of the more complex modelling, highlighting uncertainties introduced through conceptual assumptions and helping to quantify the conservatisms involved. The more stylised biosphere models are also shown capable of reproducing the results of more complex approaches. A major recommendation is that biosphere assessments need to justify the degree of complexity in modelling approaches as well as simplifying and conservative assumptions. In light of the uncertainties concerning the biosphere on very long timescales, stylised biosphere models are shown to provide a useful point of reference in themselves and remain a valuable tool for nuclear waste disposal licencing procedures. Copyright © 2015 Elsevier Ltd. All rights reserved.

Assessing the Importance of Prior Biospheric Fluxes on Inverse Model Estimates of CO2

NASA Astrophysics Data System (ADS)

Philip, S.; Johnson, M. S.; Potter, C. S.; Genovese, V. B.

2017-12-01

Atmospheric mixing ratios of carbon dioxide (CO2) are largely controlled by anthropogenic emissions and biospheric sources/sinks. The processes controlling terrestrial biosphere-atmosphere carbon exchange are currently not fully understood, resulting in models having significant differences in the quantification of biospheric CO2 fluxes. Currently, atmospheric chemical transport models (CTM) and global climate models (GCM) use multiple different biospheric CO2 flux models resulting in large differences in simulating the global carbon cycle. The Orbiting Carbon Observatory 2 (OCO-2) satellite mission was designed to allow for the improved understanding of the processes involved in the exchange of carbon between terrestrial ecosystems and the atmosphere, and therefore allowing for more accurate assessment of the seasonal/inter-annual variability of CO2. OCO-2 provides much-needed CO2 observations in data-limited regions allowing for the evaluation of model simulations of greenhouse gases (GHG) and facilitating global/regional estimates of "top-down" CO2 fluxes. We conduct a 4-D Variation (4D-Var) data assimilation with the GEOS-Chem (Goddard Earth Observation System-Chemistry) CTM using 1) OCO-2 land nadir and land glint retrievals and 2) global in situ surface flask observations to constrain biospheric CO2 fluxes. We apply different state-of-the-science year-specific CO2 flux models (e.g., NASA-CASA (NASA-Carnegie Ames Stanford Approach), CASA-GFED (Global Fire Emissions Database), Simple Biosphere Model version 4 (SiB-4), and LPJ (Lund-Postdam-Jena)) to assess the impact of "a priori" flux predictions to "a posteriori" estimates. We will present the "top-down" CO2 flux estimates for the year 2015 using OCO-2 and in situ observations, and a complete indirect evaluation of the a priori and a posteriori flux estimates using independent in situ observations. We will also present our assessment of the variability of "top-down" CO2 flux estimates when using different biospheric CO2 flux models. This work will improve our understanding of the global carbon cycle, specifically, how OCO-2 observations can be used to constrain biospheric CO2 flux model estimates.

An Estimate of the Total DNA in the Biosphere

PubMed Central

Landenmark, Hanna K. E.; Forgan, Duncan H.; Cockell, Charles S.

2015-01-01

Modern whole-organism genome analysis, in combination with biomass estimates, allows us to estimate a lower bound on the total information content in the biosphere: 5.3 × 1031 (±3.6 × 1031) megabases (Mb) of DNA. Given conservative estimates regarding DNA transcription rates, this information content suggests biosphere processing speeds exceeding yottaNOPS values (1024 Nucleotide Operations Per Second). Although prokaryotes evolved at least 3 billion years before plants and animals, we find that the information content of prokaryotes is similar to plants and animals at the present day. This information-based approach offers a new way to quantify anthropogenic and natural processes in the biosphere and its information diversity over time. PMID:26066900

An Estimate of the Total DNA in the Biosphere.

PubMed

Landenmark, Hanna K E; Forgan, Duncan H; Cockell, Charles S

2015-06-01

Modern whole-organism genome analysis, in combination with biomass estimates, allows us to estimate a lower bound on the total information content in the biosphere: 5.3 × 1031 (±3.6 × 1031) megabases (Mb) of DNA. Given conservative estimates regarding DNA transcription rates, this information content suggests biosphere processing speeds exceeding yottaNOPS values (1024 Nucleotide Operations Per Second). Although prokaryotes evolved at least 3 billion years before plants and animals, we find that the information content of prokaryotes is similar to plants and animals at the present day. This information-based approach offers a new way to quantify anthropogenic and natural processes in the biosphere and its information diversity over time.

Persistence of antibiotic-resistant and -sensitive Proteus mirabilis strains in the digestive tract of the housefly (Musca domestica) and green bottle flies (Calliphoridae).

PubMed

Wei, Ting; Miyanaga, Kazuhiko; Tanji, Yasunori

2014-10-01

Synanthropic flies have been implicated in the rapid dissemination of antibiotic-resistant bacteria and resistance determinants in the biosphere. These flies stably harbor a considerable number of bacteria that exhibit resistance to various antibiotics, but the mechanisms underlying this phenomenon remain unclear. In this study, we investigated the persistence of antibiotic-resistant bacteria in the digestive tract of houseflies and green bottle flies, using Proteus mirabilis as a model microorganism. One resistant strain carried the blaTEM and aphA1 genes, and another carried a plasmid containing qnrD gene. Quantitative PCR and 454 pyrosequencing were used to monitor the relative abundance of the Proteus strains, as well as potential changes in the overall structure of the whole bacterial community incurred by the artificial induction of Proteus cultures. Both antibiotic-resistant and -sensitive P. mirabilis strains persisted in the fly digestive tract for at least 3 days, and there was no significant difference in the relative abundance of resistant and sensitive strains despite the lower growth rate of resistant strains when cultured in vitro. Therefore, conditions in the fly digestive tract may allow resistant strains to survive the competition with sensitive strains in the absence of antibiotic selective pressure. The composition of the fly-associated bacterial community changed over time, but the contribution of the artificially introduced P. mirabilis strains to these changes was not clear. In order to explain these changes, it will be necessary to obtain more information about bacterial interspecies antagonism in the fly digestive tract.

Down under the tunic: bacterial biodiversity hotspots and widespread ammonia-oxidizing archaea in coral reef ascidians.

PubMed

Erwin, Patrick M; Pineda, Mari Carmen; Webster, Nicole; Turon, Xavier; López-Legentil, Susanna

2014-03-01

Ascidians are ecologically important components of marine ecosystems yet the ascidian microbiota remains largely unexplored beyond a few model species. We used 16S rRNA gene tag pyrosequencing to provide a comprehensive characterization of microbial symbionts in the tunic of 42 Great Barrier Reef ascidian samples representing 25 species. Results revealed high bacterial biodiversity (3 217 unique operational taxonomic units (OTU0.03) from 19 described and 14 candidate phyla) and the widespread occurrence of ammonia-oxidizing Thaumarchaeota in coral reef ascidians (24 of 25 host species). The ascidian microbiota was clearly differentiated from seawater microbial communities and included symbiont lineages shared with other invertebrate hosts as well as unique, ascidian-specific phylotypes. Several rare seawater microbes were markedly enriched (200-700 fold) in the ascidian tunic, suggesting that the rare biosphere of seawater may act as a conduit for horizontal symbiont transfer. However, most OTUs (71%) were rare and specific to single hosts and a significant correlation between host relatedness and symbiont community similarity was detected, indicating a high degree of host-specificity and potential role of vertical transmission in structuring these communities. We hypothesize that the complex ascidian microbiota revealed herein is maintained by the dynamic microenvironments within the ascidian tunic, offering optimal conditions for different metabolic pathways such as ample chemical substrate (ammonia-rich host waste) and physical habitat (high oxygen, low irradiance) for nitrification. Thus, ascidian hosts provide unique and fertile niches for diverse microorganisms and may represent an important and previously unrecognized habitat for nitrite/nitrate regeneration in coral reef ecosystems.

Strong coupling of plant and fungal community structure across western Amazonian rainforests

PubMed Central

Peay, Kabir G; Baraloto, Christopher; Fine, Paul VA

2013-01-01

The Amazon basin harbors a diverse ecological community that has a critical role in the maintenance of the biosphere. Although plant and animal communities have received much attention, basic information is lacking for fungal or prokaryotic communities. This is despite the fact that recent ecological studies have suggested a prominent role for interactions with soil fungi in structuring the diversity and abundance of tropical rainforest trees. In this study, we characterize soil fungal communities across three major tropical forest types in the western Amazon basin (terra firme, seasonally flooded and white sand) using 454 pyrosequencing. Using these data, we examine the relationship between fungal diversity and tree species richness, and between fungal community composition and tree species composition, soil environment and spatial proximity. We find that the fungal community in these ecosystems is diverse, with high degrees of spatial variability related to forest type. We also find strong correlations between α- and β-diversity of soil fungi and trees. Both fungal and plant community β-diversity were also correlated with differences in environmental conditions. The correlation between plant and fungal richness was stronger in fungal lineages known for biotrophic strategies (for example, pathogens, mycorrhizas) compared with a lineage known primarily for saprotrophy (yeasts), suggesting that this coupling is, at least in part, due to direct plant–fungal interactions. These data provide a much-needed look at an understudied dimension of the biota in an important ecosystem and supports the hypothesis that fungal communities are involved in the regulation of tropical tree diversity. PMID:23598789

Rewiring food systems to enhance human health and biosphere stewardship

NASA Astrophysics Data System (ADS)

Gordon, Line J.; Bignet, Victoria; Crona, Beatrice; Henriksson, Patrik J. G.; Van Holt, Tracy; Jonell, Malin; Lindahl, Therese; Troell, Max; Barthel, Stephan; Deutsch, Lisa; Folke, Carl; Jamila Haider, L.; Rockström, Johan; Queiroz, Cibele

2017-10-01

Food lies at the heart of both health and sustainability challenges. We use a social-ecological framework to illustrate how major changes to the volume, nutrition and safety of food systems between 1961 and today impact health and sustainability. These changes have almost halved undernutrition while doubling the proportion who are overweight. They have also resulted in reduced resilience of the biosphere, pushing four out of six analysed planetary boundaries across the safe operating space of the biosphere. Our analysis further illustrates that consumers and producers have become more distant from one another, with substantial power consolidated within a small group of key actors. Solutions include a shift from a volume-focused production system to focus on quality, nutrition, resource use efficiency, and reduced antimicrobial use. To achieve this, we need to rewire food systems in ways that enhance transparency between producers and consumers, mobilize key actors to become biosphere stewards, and re-connect people to the biosphere.

Diversity, Biogeography, and Biodegradation Potential of Actinobacteria in the Deep-Sea Sediments along the Southwest Indian Ridge

PubMed Central

Chen, Ping; Zhang, Limin; Guo, Xiaoxuan; Dai, Xin; Liu, Li; Xi, Lijun; Wang, Jian; Song, Lei; Wang, Yuezhu; Zhu, Yaxin; Huang, Li; Huang, Ying

2016-01-01

The phylum Actinobacteria has been reported to be common or even abundant in deep marine sediments, however, knowledge about the diversity, distribution, and function of actinobacteria is limited. In this study, actinobacterial diversity in the deep sea along the Southwest Indian Ridge (SWIR) was investigated using both 16S rRNA gene pyrosequencing and culture-based methods. The samples were collected at depths of 1662–4000 m below water surface. Actinobacterial sequences represented 1.2–9.1% of all microbial 16S rRNA gene amplicon sequences in each sample. A total of 5 actinobacterial classes, 17 orders, 28 families, and 52 genera were detected by pyrosequencing, dominated by the classes Acidimicrobiia and Actinobacteria. Differences in actinobacterial community compositions were found among the samples. The community structure showed significant correlations to geochemical factors, notably pH, calcium, total organic carbon, total phosphorus, and total nitrogen, rather than to spatial distance at the scale of the investigation. In addition, 176 strains of the Actinobacteria class, belonging to 9 known orders, 18 families, and 29 genera, were isolated. Among these cultivated taxa, 8 orders, 13 families, and 15 genera were also recovered by pyrosequencing. At a 97% 16S rRNA gene sequence similarity, the pyrosequencing data encompassed 77.3% of the isolates but the isolates represented only 10.3% of the actinobacterial reads. Phylogenetic analysis of all the representative actinobacterial sequences and isolates indicated that at least four new orders within the phylum Actinobacteria were detected by pyrosequencing. More than half of the isolates spanning 23 genera and all samples demonstrated activity in the degradation of refractory organics, including polycyclic aromatic hydrocarbons and polysaccharides, suggesting their potential ecological functions and biotechnological applications for carbon recycling. PMID:27621725

Results of the First Italian External Quality Assurance Scheme for somatic EGFR mutation testing in non-small-cell lung cancer.

PubMed

Normanno, Nicola; Pinto, Carmine; Taddei, Gianluigi; Gambacorta, Marcello; Castiglione, Francesca; Barberis, Massimo; Clemente, Claudio; Marchetti, Antonio

2013-06-01

The Italian Association of Medical Oncology (AIOM) and the Italian Society of Pathology and Cytology organized an external quality assessment (EQA) scheme for EGFR mutation testing in non-small-cell lung cancer. Ten specimens, including three small biopsies with known epidermal growth factor receptor (EGFR) mutation status, were validated in three referral laboratories and provided to 47 participating centers. The participants were requested to perform mutational analysis, using their usual method, and to submit results within a 4-week time frame. According to a predefined scoring system, two points were assigned to correct genotype and zero points to false-negative or false-positive results. The threshold to pass the EQA was set at higher than 18 of 20 points. Two rounds were preplanned. All participating centers submitted the results within the time frame. Polymerase chain reaction (PCR)/sequencing was the main methodology used (n = 37 laboratories), although a few centers did use pyrosequencing (n = 8) or real-time PCR (n = 2). A significant number of analytical errors were observed (n = 20), with a high frequency of false-positive results (n = 16). The lower scores were obtained for the small biopsies. Fourteen of 47 centers (30%) that did not pass the first round, having a score less than or equal to 18 points, used PCR/sequencing, whereas 10 of 10 laboratories, using pyrosequencing or real-time PCR, passed the first round. Eight laboratories passed the second round. Overall, 41of 47 centers (87%) passed the EQA. The results of the EQA for EGFR testing in non-small-cell lung cancer suggest that good quality EGFR mutational analysis is performed in Italian laboratories, although differences between testing methods were observed, especially for small biopsies.

Comparison of Bacterial Community Composition of Primary and Persistent Endodontic Infections Using Pyrosequencing.

PubMed

Tzanetakis, Giorgos N; Azcarate-Peril, M Andrea; Zachaki, Sophia; Panopoulos, Panos; Kontakiotis, Evangelos G; Madianos, Phoebus N; Divaris, Kimon

2015-08-01

Elucidating the microbial ecology of endodontic infections (EIs) is a necessary step in developing effective intracanal antimicrobials. The aim of the present study was to investigate the bacterial composition of symptomatic and asymptomatic primary and persistent infections in a Greek population using high-throughput sequencing methods. 16S amplicon pyrosequencing of 48 root canal bacterial samples was conducted, and sequencing data were analyzed using an oral microbiome-specific and a generic (Greengenes) database. Bacterial abundance and diversity were examined by EI type (primary or persistent), and statistical analysis was performed by using non-parametric and parametric tests accounting for clustered data. Bacteroidetes was the most abundant phylum in both infection groups. Significant, albeit weak associations of bacterial diversity were found, as measured by UniFrac distances with infection type (analyses of similarity, R = 0.087, P = .005) and symptoms (analyses of similarity, R = 0.055, P = .047). Persistent infections were significantly enriched for Proteobacteria and Tenericutes compared with primary ones; at the genus level, significant differences were noted for 14 taxa, including increased enrichment of persistent infections for Lactobacillus, Streptococcus, and Sphingomonas. More but less abundant phyla were identified using the Greengenes database; among those, Cyanobacteria (0.018%) and Acidobacteria (0.007%) were significantly enriched among persistent infections. Persistent infections showed higher phylogenetic diversity (PD) (asymptomatic: PD = 9.2, standard error [SE] = 1.3; symptomatic: PD = 8.2, SE = 0.7) compared with primary infections (asymptomatic: PD = 5.9, SE = 0.8; symptomatic: PD = 7.4, SE = 1.0). The present study revealed a high bacterial diversity of EI and suggests that persistent infections may have more diverse bacterial communities than primary infections. Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Response of the rare biosphere to environmental stressors in a highly diverse ecosystem (Zodletone spring, OK, USA)

PubMed Central

Coveley, Suzanne; Elshahed, Mostafa S.

2015-01-01

Within highly diverse ecosystems, the majority of bacterial taxa are present in low abundance as members of the rare biosphere. The rationale for the occurrence and maintenance of the rare biosphere, and the putative ecological role(s) and dynamics of its members within a specific ecosystem is currently debated. We hypothesized that in highly diverse ecosystems, a fraction of the rare biosphere acts as a backup system that readily responds to environmental disturbances. We tested this hypothesis by subjecting sediments from Zodletone spring, a sulfide- and sulfur-rich spring in Southwestern OK, to incremental levels of salinity (1, 2, 3, 4, and 10% NaCl), or temperature (28°, 30°, 32°, and 70 °C), and traced the trajectories of rare members of the community in response to these manipulations using 16S rRNA gene analysis. Our results indicate that multiple rare bacterial taxa are promoted from rare to abundant members of the community following such manipulations and that, in general, the magnitude of such recruitment is directly proportional to the severity of the applied manipulation. Rare members that are phylogenetically distinct from abundant taxa in the original sample (unique rare biosphere) played a more important role in the microbial community response to environmental disturbances, compared to rare members that are phylogenetically similar to abundant taxa in the original sample (non-unique rare biosphere). The results emphasize the dynamic nature of the rare biosphere, and highlight its complexity and non-monolithic nature. PMID:26312178

Prokaryotic microbiota in the digestive cavity of the jellyfish Cotylorhiza tuberculata.

PubMed

Cortés-Lara, Sara; Urdiain, Mercedes; Mora-Ruiz, Merit; Prieto, Laura; Rosselló-Móra, Ramon

2015-10-01

The microbiota associated to the gastric cavity of four exemplars of the jellyfish Cotylorhiza tuberculata has been studied by means of cultured-dependent and -independent methods. The pyrosequencing approach rendered a very reduced diversity of Bacteria with four major groups shared by the four exemplars that made up to 95% of the total diversity. The culturing approach recovered low abundant organisms and some of them also detected by the pyrosequencing approach. The major key organisms were related to the genera Spiroplasma, Thalassospira, Tenacibaculum (from the pyrosequencing data), and Vibrio (from the cultivable fraction). Altogether the results indicate that C. tuberculata harbors an associated microbiota of very reduced diversity. On the other hand, some of the major key players may be potential pathogens and the host may serve as dispersal mechanism. Copyright © 2015 Elsevier GmbH. All rights reserved.

Comparing Amazon Basin CO2 fluxes from an atmospheric inversion with TRENDY biosphere models

NASA Astrophysics Data System (ADS)

Diffenbaugh, N. S.; Alden, C. B.; Harper, A. B.; Ahlström, A.; Touma, D. E.; Miller, J. B.; Gatti, L. V.; Gloor, M.

2015-12-01

Net exchange of carbon dioxide (CO2) between the atmosphere and the terrestrial biosphere is sensitive to environmental conditions, including extreme heat and drought. Of particular importance for local and global carbon balance and climate are the expansive tracts of tropical rainforest located in the Amazon Basin. Because of the Basin's size and ecological heterogeneity, net biosphere CO2 exchange with the atmosphere remains largely un-constrained. In particular, the response of net CO2 exchange to changes in environmental conditions such as temperature and precipitation are not yet well known. However, proper representation of these relationships in biosphere models is a necessary constraint for accurately modeling future climate and climate-carbon cycle feedbacks. In an effort to compare biosphere response to climate across different biosphere models, the TRENDY model intercomparison project coordinated the simulation of CO2 fluxes between the biosphere and atmosphere, in response to historical climate forcing, by 9 different Dynamic Global Vegetation Models. We examine the TRENDY model results in the Amazon Basin, and compare this "bottom-up" method with fluxes derived from a "top-down" approach to estimating net CO2 fluxes, obtained through atmospheric inverse modeling using CO2 measurements sampled by aircraft above the basin. We compare the "bottom-up" and "top-down" fluxes in 5 sub-regions of the Amazon basin on a monthly basis for 2010-2012. Our results show important periods of agreement between some models in the TRENDY suite and atmospheric inverse model results, notably the simulation of increased biosphere CO2 loss during wet season heat in the Central Amazon. During the dry season, however, model ability to simulate observed response of net CO2 exchange to drought was varied, with few models able to reproduce the "top-down" inversion flux signals. Our results highlight the value of atmospheric trace gas observations for helping to narrow the possibilities of future carbon-climate interactions, especially in historically under-observed regions like the Amazon.

Application of the Spanish methodological approach for biosphere assessment to a generic high-level waste disposal site.

PubMed

Agüero, A; Pinedo, P; Simón, I; Cancio, D; Moraleda, M; Trueba, C; Pérez-Sánchez, D

2008-09-15

A methodological approach which includes conceptual developments, methodological aspects and software tools have been developed in the Spanish context, based on the BIOMASS "Reference Biospheres Methodology". The biosphere assessments have to be undertaken with the aim of demonstrating compliance with principles and regulations established to limit the possible radiological impact of radioactive waste disposals on human health and on the environment, and to ensure that future generations will not be exposed to higher radiation levels than those that would be acceptable today. The biosphere in the context of high-level waste disposal is defined as the collection of various radionuclide transfer pathways that may result in releases into the surface environment, transport within and between the biosphere receptors, exposure of humans and biota, and the doses/risks associated with such exposures. The assessments need to take into account the complexity of the biosphere, the nature of the radionuclides released and the long timescales considered. It is also necessary to make assumptions related to the habits and lifestyle of the exposed population, human activities in the long term and possible modifications of the biosphere. A summary on the Spanish methodological approach for biosphere assessment are presented here as well as its application in a Spanish generic case study. A reference scenario has been developed based on current conditions at a site located in Central-West Spain, to indicate the potential impact to the actual population. In addition, environmental change has been considered qualitatively through the use of interaction matrices and transition diagrams. Unit source terms of (36)Cl, (79)Se, (99)Tc, (129)I, (135)Cs, (226)Ra, (231)Pa, (238)U, (237)Np and (239)Pu have been taken. Two exposure groups of infants and adults have been chosen for dose calculations. Results are presented and their robustness is evaluated through the use of uncertainty and sensitivity analyses.

Piezophilic Bacteria Isolated from Sediment of the Shimokita Coalbed, Japan

NASA Astrophysics Data System (ADS)

Fang, J.; Kato, C.; Hori, T.; Morono, Y.; Inagaki, F.

2013-12-01

The Earth is a cold planet as well as pressured planet, hosting both the surface biosphere and the deep biosphere. Pressure ranges over four-orders of magnitude in the surface biosphere and probably more in the deep biosphere. Pressure is an important thermodynamic property of the deep biosphere that affects microbial physiology and biochemistry. Bacteria that require high-pressure conditions for optimal growth are called piezophilic bacteria. Subseafloor marine sediments are one of the most extensive microbial habitats on Earth. Marine sediments cover more than two-thirds of the Earth's surface, and represent a major part of the deep biosphere. Owing to its vast size and intimate connection with the surface biosphere, particularly the oceans, the deep biosphere has enormous potential for influencing global-scale biogeochemical processes, including energy, climate, carbon and nutrient cycles. Therefore, studying piezophilic bacteria of the deep biosphere has important implications in increasing our understanding of global biogeochemical cycles, the interactions between the biosphere and the geosphere, and the evolution of life. Sediment samples were obtained during IODP Expedition 337, from 1498 meters below sea floor (mbsf) (Sample 6R-3), 1951~1999 mbsf (19R-1~25R-3; coalbed mix), and 2406 mbsf (29R-7). The samples were mixed with MB2216 growth medium and cultivated under anaerobic conditions at 35 MPa (megapascal) pressure. Growth temperatures were adjusted to in situ environmental conditions, 35°C for 6R-3, 45°C for 19R-1~25R-3, and 55°C for 29R-7. The cultivation was performed three times, for 30 days each time. Microbial cells were obtained and the total DNA was extracted. At the same time, isolation of microbes was also performed under anaerobic conditions. Microbial communities in the coalbed sediment were analyzed by cloning, sequencing, and terminal restriction fragment length polymorphism (t-RFLP) of 16S ribosomal RNA genes. From the partial 16S rRNA gene sequences, we have identified abundant Alkalibacterium sp. in 6R-3 and 29R-7 at the first HP cultivation. We also identified Haloactibacillus sp. in 6R-3 and Anoxybacillus related sp. in 19R-1~25R-3 at the third HP cultivation. These microorganisms are likely piezophiles and play an important role in degradation of sedimentary organic matter and production of microbial metabolites sustaining the deep microbial ecosystem in the Shimokita Coalbed. The complete 16S sequencing and isolation of piezophiles are now ongoing.

Descriptive Biomarkers for Assessing Breast Cancer Risk

DTIC Science & Technology

2010-10-01

and we are making significant progress on Tasks 6 and 7. We completed methylation analyses of three genes (RASSF1, SFRP1 and GSTP1 ) on all samples...promoter hypermethylation; RASSF1, GSTP1 , SFRP1 12 karcaro@nre.umass.edu Arcaro, Kathleen F Annual Report...methylation analysis by pyrosequencing. PCR amplification and pyrosequencing has been completed for three genes, RASSF1, SFRP1 and GSTP1 and have

Pyrosequencing Analysis of Norovirus Genogroup II Distribution in Sewage and Oysters: First Detection of GII.17 Kawasaki 2014 in Oysters.

PubMed

Pu, Jian; Kazama, Shinobu; Miura, Takayuki; Azraini, Nabila Dhyan; Konta, Yoshimitsu; Ito, Hiroaki; Ueki, You; Cahyaningrum, Ermaya Eka; Omura, Tatsuo; Watanabe, Toru

2016-12-01

Norovirus GII.3, GII.4, and GII.17 were detected using pyrosequencing in sewage and oysters in January and February 2015, in Japan. The strains in sewage and oyster samples were genetically identical or similar, predominant strains belonging to GII.17 Kawasaki 2014 lineage. This is the first report of GII.17 Kawasaki 2014 in oysters.

Combined Real-Time PCR and Pyrosequencing Strategy for Objective, Sensitive, Specific, and High-Throughput Identification of Reduced Susceptibility to Penicillins in Neisseria meningitidis▿

PubMed Central

Thulin, Sara; Olcén, Per; Fredlund, Hans; Unemo, Magnus

2008-01-01

A segment of penA in Neisseria meningitidis strains (n = 127), including two nucleotide sites closely associated to reduced susceptibility to penicillins, was amplified and pyrosequenced. All results were in concordance with Sanger sequencing, and a high correlation between alterations in the two Peni-specific sites and reduced susceptibility to penicillins was identified. PMID:18070955

Mitochondrial sequence analysis for forensic identification using pyrosequencing technology.

PubMed

Andréasson, H; Asp, A; Alderborn, A; Gyllensten, U; Allen, M

2002-01-01

Over recent years, requests for mtDNA analysis in the field of forensic medicine have notably increased, and the results of such analyses have proved to be very useful in forensic cases where nuclear DNA analysis cannot be performed. Traditionally, mtDNA has been analyzed by DNA sequencing of the two hypervariable regions, HVI and HVII, in the D-loop. DNA sequence analysis using the conventional Sanger sequencing is very robust but time consuming and labor intensive. By contrast, mtDNA analysis based on the pyrosequencing technology provides fast and accurate results from the human mtDNA present in many types of evidence materials in forensic casework. The assay has been developed to determine polymorphic sites in the mitochondrial D-loop as well as the coding region to further increase the discrimination power of mtDNA analysis. The pyrosequencing technology for analysis of mtDNA polymorphisms has been tested with regard to sensitivity, reproducibility, and success rate when applied to control samples and actual casework materials. The results show that the method is very accurate and sensitive; the results are easily interpreted and provide a high success rate on casework samples. The panel of pyrosequencing reactions for the mtDNA polymorphisms were chosen to result in an optimal discrimination power in relation to the number of bases determined.

Identification and genotyping of molluscum contagiosum virus from genital swab samples by real-time PCR and Pyrosequencing.

PubMed

Trama, Jason P; Adelson, Martin E; Mordechai, Eli

2007-12-01

Laboratory diagnosis of molluscum contagiosum virus (MCV) is important as lesions can be confused with those caused by Cryptococcus neoformans, herpes simplex virus, human papillomavirus, and varicella-zoster virus. To develop a rapid method for identifying patients infected with MCV via swab sampling. Two dual-labeled probe real-time PCR assays, one homologous to the p43K gene and one to the MC080R gene, were designed. The p43K PCR was designed to be used in conjunction with Pyrosequencing for confirmation of PCR products and discrimination between MCV1 and MCV2. Both PCR assays were optimized with respect to reaction components, thermocycling parameters, and primer and probe concentrations. The specificities of both PCR assays were confirmed by non-amplification of 38 known human pathogens. Sensitivity assays demonstrated detection of as few as 10 copies per reaction. Testing 703 swabs, concordance between the two real-time PCR assays was 99.9%. Under the developed conditions, Pyrosequencing of the p43K PCR product was capable of providing enough nucleotide sequence to definitively differentiate MCV1 and MCV2. These real-time PCR assays can be used for the rapid, sensitive, and specific detection of MCV and, when combined with Pyrosequencing, can further discriminate between MCV1 and MCV2.

Microbial Diversity Analysis of Fermented Mung Beans (Lu-Doh-Huang) by Using Pyrosequencing and Culture Methods

PubMed Central

Chao, Shiou-Huei; Huang, Hui-Yu; Chang, Chuan-Hsiung; Yang, Chih-Hsien; Cheng, Wei-Shen; Kang, Ya-Huei; Watanabe, Koichi; Tsai, Ying-Chieh

2013-01-01

In Taiwanese alternative medicine Lu-doh-huang (also called Pracparatum mungo), mung beans are mixed with various herbal medicines and undergo a 4-stage process of anaerobic fermentation. Here we used high-throughput sequencing of the 16S rRNA gene to profile the bacterial community structure of Lu-doh-huang samples. Pyrosequencing of samples obtained at 7 points during fermentation revealed 9 phyla, 264 genera, and 586 species of bacteria. While mung beans were inside bamboo sections (stages 1 and 2 of the fermentation process), family Lactobacillaceae and genus Lactobacillus emerged in highest abundance; Lactobacillus plantarum was broadly distributed among these samples. During stage 3, the bacterial distribution shifted to family Porphyromonadaceae, and Butyricimonas virosa became the predominant microbial component. Thereafter, bacterial counts decreased dramatically, and organisms were too few to be detected during stage 4. In addition, the microbial compositions of the liquids used for soaking bamboo sections were dramatically different: Exiguobacterium mexicanum predominated in the fermented soybean solution whereas B. virosa was predominant in running spring water. Furthermore, our results from pyrosequencing paralleled those we obtained by using the traditional culture method, which targets lactic acid bacteria. In conclusion, the microbial communities during Lu-doh-huang fermentation were markedly diverse, and pyrosequencing revealed a complete picture of the microbial consortium. PMID:23700436

Earth's early biosphere

NASA Technical Reports Server (NTRS)

Des Marais, D. J.

1998-01-01

Understanding our own early biosphere is essential to our search for life elsewhere, because life arose on Earth very early and rocky planets shared similar early histories. The biosphere arose before 3.8 Ga ago, was exclusively unicellular and was dominated by hyperthermophiles that utilized chemical sources of energy and employed a range of metabolic pathways for CO2 assimilation. Photosynthesis also arose very early. Oxygenic photosynthesis arose later but still prior to 2.7 Ga. The transition toward the modern global environment was paced by a decline in volcanic and hydrothermal activity. These developments allowed atmospheric O2 levels to increase. The O2 increase created new niches for aerobic life, most notably the more advanced Eukarya that eventually spawned the megascopic fauna and flora of our modern biosphere.

Recent developments in assessment of long-term radionuclide behavior in the geosphere-biosphere subsystem.

PubMed

Smith, G M; Smith, K L; Kowe, R; Pérez-Sánchez, D; Thorne, M; Thiry, Y; Read, D; Molinero, J

2014-05-01

Decisions on permitting, controlling and monitoring releases of radioactivity into the environment rely on a great variety of factors. Important among these is the prospective assessment of radionuclide behavior in the environment, including migration and accumulation among and within specific environmental media, and the resulting environmental and human health impacts. Models and techniques to undertake such assessments have been developed over several decades based on knowledge of the ecosystems involved, as well as monitoring of previous radionuclide releases to the environment, laboratory experiments and other related research. This paper presents developments in the assessment of radiation doses and related research for some of the key radionuclides identified as of potential significance in the context of releases to the biosphere from disposal facilities for solid radioactive waste. Since releases to the biosphere from disposal facilities involve transfers from the geosphere to the biosphere, an important aspect is the combined effects of surface hydrology, near-surface hydrogeology and chemical gradients on speciation and radionuclide mobility in the zone in which the geosphere and biosphere overlap (herein described as the geosphere-biosphere subsystem). In turn, these aspects of the environment can be modified as a result of environmental change over the thousands of years that have to be considered in radioactive waste disposal safety assessments. Building on the experience from improved understanding of the behavior of the key radionuclides, this paper proceeds to describe development of a generic methodology for representing the processes and environmental changes that are characteristic of the interface between the geosphere and the biosphere. The information that is provided and the methodology that is described are based on international collaborative work implemented through the BIOPROTA forum, www.bioprota.org. Copyright © 2013 Elsevier Ltd. All rights reserved.

Rapid Identification of Genetic Modifications in Bacillus anthracis Using Whole Genome Draft Sequences Generated by 454 Pyrosequencing

DTIC Science & Technology

2010-08-25

or intentional genetic modifications that circumvent the targets of the detection assays or in the case of a biological attack using an antibiotic ...genetic changes conferring antibiotic resistance can be deciphered rapidly and accurately using WGS. We demonstrate the utility of Roche 454...Rapid Identification of Genetic Modifications in Bacillus anthracis Using Whole Genome Draft Sequences Generated by 454 Pyrosequencing Peter E. Chen1

The UNESCO biosphere reserve concept as a tool for urban sustainability: the CUBES Cape Town case study.

PubMed

Stanvliet, R; Jackson, J; Davis, G; De Swardt, C; Mokhoele, J; Thom, Q; Lane, B D

2004-06-01

The Cape Town Case Study (CTCS) was a multi-institutional collaborative project initiated by CUBES, a knowledge networking initiative of UNESCO's Ecological Sciences Division and the Earth Institute at Columbia University. Cape Town was selected as a CUBES site on the basis of its high biological and cultural significance, together with its demonstrated leadership in promoting urban sustainability. The CTCS was conducted by the Cape Town Urban Biosphere Group, a cross-disciplinary group of specialists drawn from national, provincial, municipal, and civil society institutions, mandated to examine the potential value of the UNESCO Biosphere Reserve concept as a tool for environmental management, social inclusion, and poverty alleviation in Cape Town. This article provides a contextualization of the CTCS and its collaborative process. It also reviews the biosphere reserve concept relative to urban sustainability objectives and proposes a more functional application of that concept in an urban context. A detailed analysis of key initiatives at the interface of conservation and poverty alleviation is provided in table format. Drawing on an examination of successful sustainability initiatives in Cape Town, specific recommendations are made for future application of the biosphere reserve concept in an urban context, as well as a model by which urban areas might affiliate with the UNESCO World Network of Biosphere Reserves, and criteria for such affiliation.

Lignocellulose deconstruction in the biosphere

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bomble, Yannick J.; Lin, Chien-Yuan; Amore, Antonella

Microorganisms have evolved different and yet complementary mechanisms to degrade biomass in the biosphere. The chemical biology of lignocellulose deconstruction is a complex and intricate process that appears to vary in response to specific ecosystems. These microorganisms rely on simple to complex arrangements of glycoside hydrolases to conduct most of these polysaccharide depolymerization reactions and also, as discovered more recently, oxidative mechanisms via lytic polysaccharide monooxygenases or non-enzymatic Fenton reactions which are used to enhance deconstruction. It is now clear that these deconstruction mechanisms are often more efficient in the presence of the microorganisms. In general, a major fraction ofmore » the total plant biomass deconstruction in the biosphere results from the action of various microorganisms, primarily aerobic bacteria and fungi, as well as a variety of anaerobic bacteria. Beyond carbon recycling, specialized microorganisms interact with plants to manage nitrogen in the biosphere. Understanding the interplay between these organisms within or across ecosystems is crucial to further our grasp of chemical recycling in the biosphere and also enables optimization of the burgeoning plant-based bioeconomy.« less

Top–down assessment of the Asian carbon budget since the mid 1990s

PubMed Central

Thompson, R. L.; Patra, P. K.; Chevallier, F.; Maksyutov, S.; Law, R. M.; Ziehn, T.; van der Laan-Luijkx, I. T.; Peters, W.; Ganshin, A.; Zhuravlev, R.; Maki, T.; Nakamura, T.; Shirai, T.; Ishizawa, M.; Saeki, T.; Machida, T.; Poulter, B.; Canadell, J. G.; Ciais, P.

2016-01-01

Increasing atmospheric carbon dioxide (CO2) is the principal driver of anthropogenic climate change. Asia is an important region for the global carbon budget, with 4 of the world's 10 largest national emitters of CO2. Using an ensemble of seven atmospheric inverse systems, we estimated land biosphere fluxes (natural, land-use change and fires) based on atmospheric observations of CO2 concentration. The Asian land biosphere was a net sink of −0.46 (−0.70–0.24) PgC per year (median and range) for 1996–2012 and was mostly located in East Asia, while in South and Southeast Asia the land biosphere was close to carbon neutral. In East Asia, the annual CO2 sink increased between 1996–2001 and 2008–2012 by 0.56 (0.30–0.81) PgC, accounting for ∼35% of the increase in the global land biosphere sink. Uncertainty in the fossil fuel emissions contributes significantly (32%) to the uncertainty in land biosphere sink change. PMID:26911442

Lignocellulose deconstruction in the biosphere.

PubMed

Bomble, Yannick J; Lin, Chien-Yuan; Amore, Antonella; Wei, Hui; Holwerda, Evert K; Ciesielski, Peter N; Donohoe, Bryon S; Decker, Stephen R; Lynd, Lee R; Himmel, Michael E

2017-12-01

Microorganisms have evolved different and yet complementary mechanisms to degrade biomass in the biosphere. The chemical biology of lignocellulose deconstruction is a complex and intricate process that appears to vary in response to specific ecosystems. These microorganisms rely on simple to complex arrangements of glycoside hydrolases to conduct most of these polysaccharide depolymerization reactions and also, as discovered more recently, oxidative mechanisms via lytic polysaccharide monooxygenases or non-enzymatic Fenton reactions which are used to enhance deconstruction. It is now clear that these deconstruction mechanisms are often more efficient in the presence of the microorganisms. In general, a major fraction of the total plant biomass deconstruction in the biosphere results from the action of various microorganisms, primarily aerobic bacteria and fungi, as well as a variety of anaerobic bacteria. Beyond carbon recycling, specialized microorganisms interact with plants to manage nitrogen in the biosphere. Understanding the interplay between these organisms within or across ecosystems is crucial to further our grasp of chemical recycling in the biosphere and also enables optimization of the burgeoning plant-based bioeconomy. Copyright © 2017. Published by Elsevier Ltd.

Analysis of Low-Biomass Microbial Communities in the Deep Biosphere.

PubMed

Morono, Y; Inagaki, F

2016-01-01

Over the past few decades, the subseafloor biosphere has been explored by scientific ocean drilling to depths of about 2.5km below the seafloor. Although organic-rich anaerobic sedimentary habitats in the ocean margins harbor large numbers of microbial cells, microbial populations in ultraoligotrophic aerobic sedimentary habitats in the open ocean gyres are several orders of magnitude less abundant. Despite advances in cultivation-independent molecular ecological techniques, exploring the low-biomass environment remains technologically challenging, especially in the deep subseafloor biosphere. Reviewing the historical background of deep-biosphere analytical methods, the importance of obtaining clean samples and tracing contamination, as well as methods for detecting microbial life, technological aspects of molecular microbiology, and detecting subseafloor metabolic activity will be discussed. Copyright © 2016 Elsevier Inc. All rights reserved.

Biosphere2 and Earthbuzz

NASA Astrophysics Data System (ADS)

Washburne, J. C.

2009-12-01

In an attempt to reach a broader audience, Biosphere 2, near Tucson, AZ, is participating in a network of science centers thanks to new funding through the Science Museum of Minnesota (SMM) and the National Center for Earth System Dynamics (NCED). Each of these centers will be tied together through an Earthbuzz kiosk, basically a networked web site that allows visitors to learn more about the work of leading local scientists in a very personal and captivating format. Content is currently being developed by Biosphere 2 researchers, staff, and graduate students that range from a public question and answer forum called “Scientist on the Spot” to science blogs by Biosphere 2 Fellows. It is hoped that this project will help educate the public about the Anthropocene, that is, the current geologic period that is so greatly affected by humankind’s impact on the health of the planet. Biosphere 2 provides a unique location to engage the public in this conversation for several reasons. First, no other destination on Earth gives the public such a physical immersion into what climate change might mean as does Biosphere 2. On the regular walking tour, visitors are guided through scaled down versions of an African savannah, a semi-arid thorn scrub, a coastal fog desert and a tropical rainforest. Digital displays of temperature and humidity confirm what your body is feeling - conditions ranging from desert aridity to tropical humidity. As one passes through the biomes of Biosphere 2, climate change is a whole body experience. Second, Biosphere 2 is also an active ecological research site - part of a unique network of sites run by the University of Arizona that allow scientists to study ecosystem processes across a range of scales - from microscopic root studies to studies encompassing large watersheds. In particular, a group of researchers is studying why large stands of pinion-juniper forests across the southwest have died in recent years. Biosphere2’s role in this experiment is multi-faceted. Boxed Pinion pines have been placed in a setting that allows close regulation of both temperature and precipitation while being intensively monitored electronically. Graduate students are learning field protocols in close proximity to their classes and are encouraged to interact with the almost hourly public tours. While many scientists are able to wow their peers at AGU, it is quite another thing to assess the ever changing mix of ages and interests present in the tour groups and to clearly communicate your scientific objectives and to explain what you happen to be working on today. These and many other examples will be highlighted in this presentation of how Biosphere 2 is developing an Earthbuzz kiosk in conjunction with the SMM and NCED.

Multiplex pyrosequencing assay using AdvISER-MH-PYRO algorithm: a case for rapid and cost-effective genotyping analysis of prostate cancer risk-associated SNPs.

PubMed

Ambroise, Jérôme; Butoescu, Valentina; Robert, Annie; Tombal, Bertrand; Gala, Jean-Luc

2015-06-25

Single Nucleotide Polymorphisms (SNPs) identified in Genome Wide Association Studies (GWAS) have generally moderate association with related complex diseases. Accordingly, Multilocus Genetic Risk Scores (MGRSs) have been computed in previous studies in order to assess the cumulative association of multiple SNPs. When several SNPs have to be genotyped for each patient, using successive uniplex pyrosequencing reactions increases analytical reagent expenses and Turnaround Time (TAT). While a set of several pyrosequencing primers could theoretically be used to analyze multiplex amplicons, this would generate overlapping primer-specific pyro-signals that are visually uninterpretable. In the current study, two multiplex assays were developed consisting of a quadruplex (n=4) and a quintuplex (n=5) polymerase chain reaction (PCR) each followed by multiplex pyrosequencing analysis. The aim was to reliably but rapidly genotype a set of prostate cancer-related SNPs (n=9). The nucleotide dispensation order was selected using SENATOR software. Multiplex pyro-signals were analyzed using the new AdvISER-MH-PYRO software based on a sparse representation of the signal. Using uniplex assays as gold standard, the concordance between multiplex and uniplex assays was assessed on DNA extracted from patient blood samples (n = 10). All genotypes (n=90) generated with the quadruplex and the quintuplex pyroquencing assays were perfectly (100 %) concordant with uniplex pyrosequencing. Using multiplex genotyping approach for analyzing a set of 90 patients allowed reducing TAT by approximately 75 % (i.e., from 2025 to 470 min) while reducing reagent consumption and cost by approximately 70 % (i.e., from ~229 US$ /patient to ~64 US$ /patient). This combination of quadruplex and quintuplex pyrosequencing and PCR assays enabled to reduce the amount of DNA required for multi-SNP analysis, and to lower the global TAT and costs of SNP genotyping while providing results as reliable as uniplex analysis. Using this combined multiplex approach also substantially reduced the production of waste material. These genotyping assays appear therefore to be biologically, economically and ecologically highly relevant, being worth to be integrated in genetic-based predictive strategies for better selecting patients at risk for prostate cancer. In addition, the same approach could now equally be transposed to other clinical/research applications relying on the computation of MGRS based on multi-SNP genotyping.

Prebiological Synthesis Organic Matter and Origin of Life in Protoplanetary Disks

NASA Astrophysics Data System (ADS)

Snytnikov, V. N.

2017-05-01

Processes and conditions around new born Sun and possibly other young stars led to the origin of life in the circumstellar disc in a few million years. Than the circumsolar biosphere evolved to its current earth state. In agreement with the main results of the natural sciences and the theory of self-organization, several stages are necessary for the life to emerge on the Earth. Nowday we can specify "The cold prehistiry of the life", "RNA-world", "Preplanet biosphere", "Destructive biosphere", "Earth of bacteria".

Use of midlatitude soil moisture and meteorological observations to validate soil moisture simulations with biosphere and bucket models

NASA Technical Reports Server (NTRS)

Robock, Alan; Vinnikov, Konstantin YA.; Schlosser, C. Adam; Speranskaya, Nina A.; Xue, Yongkang

1995-01-01

Soil moisture observations in sites with natural vegetation were made for several decades in the former Soviet Union at hundreds of stations. In this paper, the authors use data from six of these stations from different climatic regimes, along with ancillary meteorological and actinometric data, to demonstrate a method to validate soil moisture simulations with biosphere and bucket models. Some early and current general circulation models (GCMs) use bucket models for soil hydrology calculations. More recently, the Simple Biosphere Model (SiB) was developed to incorporate the effects of vegetation on fluxes of moisture, momentum, and energy at the earth's surface into soil hydrology models. Until now, the bucket and SiB have been verified by comparison with actual soil moisture data only on a limited basis. In this study, a Simplified SiB (SSiB) soil hydrology model and a 15-cm bucket model are forced by observed meteorological and actinometric data every 3 h for 6-yr simulations at the six stations. The model calculations of soil moisture are compared to observations of soil moisture, literally 'ground truth,' snow cover, surface albedo, and net radiation, and with each other. For three of the stations, the SSiB and 15-cm bucket models produce good simulations of seasonal cycles and interannual variations of soil moisture. For the other three stations, there are large errors in the simulations by both models. Inconsistencies in specification of field capacity may be partly responsible. There is no evidence that the SSiB simulations are superior in simulating soil moisture variations. In fact, the models are quite similar since SSiB implicitly has a bucket embedded in it. One of the main differences between the models is in the treatment of runoff due to melting snow in the spring -- SSiB incorrectly puts all the snowmelt into runoff. While producing similar soil moisture simulations, the models produce very different surface latent and sensible heat fluxes, which would have large effects on GCM simulations.

The water cycle in closed ecological systems: perspectives from the Biosphere 2 and Laboratory Biosphere systems

NASA Astrophysics Data System (ADS)

Nelson, Mark; Dempster, William; Allen, John P.

To achieve sustainable and healthy closed ecological systems requires successful solutions to the challenge of closing the water cycle - recycling wastewater/soil leachate and evaporateed water and supplying water of required quality as needed for different needs within the facility. Engineering Biosphere 2, the first multi-biome closed ecological system, total footprint of the airtight area is 12,700 m2 with a combined volume of 200,000 m3 with a total water capacity of some 6 x 106 liters of water presented a complex challenge because it included human inhabitants, their agricultural and technical systems, as well as a range of analogue ecosystems ranging from rainforest to desert, freshwater ecologies to mini-ocean coral reef ecosystems. By contrast, the Laboratory Biosphere - a small (40m3 volume) soil-based plant growth facility with a footprint of 15m3 - is a very simplified system, but with some similar issues such as salinity management and the provision of water quality sufficient for plant growth. In Biosphere 2, water needs included supplying potable water for people and domestic animals, irrigation water for a wide variety of food crops, and recycling and recovering soil nutrients from wastewater. In the wilderness biomes, adequately freshwater was needed for terrestrial ecosystems and maintaining appropriate salinity and pH in aquatic/marine ecosystems. The largest reservoirs in Biosphere 2 were the ocean/marsh with some 4x106 liters, soil with 2 x 106 liters, primary storage tanks with a capacity for up to 8 x 105 liters and storage tanks for condensate collection and mixing tanks with 1.5 x 105 liters to supply irrigation for farm and wilderness ecosystems. Other reservoirs were far smaller - humidity in the atmosphere (2 x 103 liters), streams in the rainforest and savannah, and seasonal pools in the desert were orders of magnitude smaller (8 x 103 liters). Key technologies included condensation from humidity in the airhandlers and from the glass spaceframe to produce high quality freshwater, purification of nutrients from ocean waters with first algae mats and then protein skimmers, wastewater treatment with constructed wetlands and desalination through reverse osmosis and flash evaporation were key to recycling water with appropriate quality throughout the Biosphere 2 facility. Wastewater from all human uses and the domestic animals in Biosphere 2 was treated and recycling through a series of constructed wetlands, which had hydraulic loading of 0.9-1.1 m3 day-1 (240-290 gal d-1). Plant production in the wetland treatment system produced 1210 kg dry weight of emergent and floating aquatic plant wetland used as fodder for the domestic animals and remaining nutrients/water was reused as part of the agricultural irrigation supply. There were pools of water with recycling times of days to weeks and others with far longer cycling times within Biosphere 2. By contrast, the Laboratory Biosphere with a total water reservoir of less than 500 litres has far quicker cycling rapidity. However, just as in Biosphere 2, humidity in the Laboratory Biosphere is a very small reservoir of water. The amount of water passing through the air in the course of a 12-hour operational day is two orders of magnitude greater than the amount stored in the air. Thus evaporation, condensation and soil leachate collection are vital parts of the recycle system just as in Biosphere 2. The water cycle and sustainable water recycling in closed ecological systems presents problems requiring further research to resolve - such as how to control buildup of salinity in materially-closed ecosystems and effective ways to retain nutrients in optimal quantity and useable form for plant growth which are common to closed ecological systems of whatever size. These issues have relevance to a global environment increasingly facing water shortages and the task of maintaining water quality for human and ecosystem health.

Fungal Diversity in Tomato Rhizosphere Soil under Conventional and Desert Farming Systems

PubMed Central

Kazerooni, Elham A.; Maharachchikumbura, Sajeewa S. N.; Rethinasamy, Velazhahan; Al-Mahrouqi, Hamed; Al-Sadi, Abdullah M.

2017-01-01

This study examined fungal diversity and composition in conventional (CM) and desert farming (DE) systems in Oman. Fungal diversity in the rhizosphere of tomato was assessed using 454-pyrosequencing and culture-based techniques. Both techniques produced variable results in terms of fungal diversity, with 25% of the fungal classes shared between the two techniques. In addition, pyrosequencing recovered more taxa compared to direct plating. These findings could be attributed to the ability of pyrosequencing to recover taxa that cannot grow or are slow growing on culture media. Both techniques showed that fungal diversity in the conventional farm was comparable to that in the desert farm. However, the composition of fungal classes and taxa in the two farming systems were different. Pyrosequencing revealed that Microsporidetes and Dothideomycetes are the two most common fungal classes in CM and DE, respectively. However, the culture-based technique revealed that Eurotiomycetes was the most abundant class in both farming systems and some classes, such as Microsporidetes, were not detected by the culture-based technique. Although some plant pathogens (e.g., Pythium or Fusarium) were detected in the rhizosphere of tomato, the majority of fungal species in the rhizosphere of tomato were saprophytes. Our study shows that the cultivation system may have an impact on fungal diversity. The factors which affected fungal diversity in both farms are discussed. PMID:28824590

A comprehensive insight into bacterial virulence in drinking water using 454 pyrosequencing and Illumina high-throughput sequencing.

PubMed

Huang, Kailong; Zhang, Xu-Xiang; Shi, Peng; Wu, Bing; Ren, Hongqiang

2014-11-01

In order to comprehensively investigate bacterial virulence in drinking water, 454 pyrosequencing and Illumina high-throughput sequencing were used to detect potential pathogenic bacteria and virulence factors (VFs) in a full-scale drinking water treatment and distribution system. 16S rRNA gene pyrosequencing revealed high bacterial diversity in the drinking water (441-586 operational taxonomic units). Bacterial diversity decreased after chlorine disinfection, but increased after pipeline distribution. α-Proteobacteria was the most dominant taxonomic class. Alignment against the established pathogen database showed that several types of putative pathogens were present in the drinking water and Pseudomonas aeruginosa had the highest abundance (over 11‰ of total sequencing reads). Many pathogens disappeared after chlorine disinfection, but P. aeruginosa and Leptospira interrogans were still detected in the tap water. High-throughput sequencing revealed prevalence of various pathogenicity islands and virulence proteins in the drinking water, and translocases, transposons, Clp proteases and flagellar motor switch proteins were the predominant VFs. Both diversity and abundance of the detectable VFs increased after the chlorination, and decreased after the pipeline distribution. This study indicates that joint use of 454 pyrosequencing and Illumina sequencing can comprehensively characterize environmental pathogenesis, and several types of putative pathogens and various VFs are prevalent in drinking water. Copyright © 2014 Elsevier Inc. All rights reserved.

Detection of novel NF1 mutations and rapid mutation prescreening with Pyrosequencing.

PubMed

Brinckmann, Anja; Mischung, Claudia; Bässmann, Ingelore; Kühnisch, Jirko; Schuelke, Markus; Tinschert, Sigrid; Nürnberg, Peter

2007-12-01

Neurofibromatosis type 1 (NF1) is caused by mutations in the neurofibromin (NF1) gene. Mutation analysis of NF1 is complicated by its large size, the lack of mutation hotspots, pseudogenes and frequent de novo mutations. Additionally, the search for NF1 mutations on the mRNA level is often hampered by nonsense-mediated mRNA decay (NMD) of the mutant allele. In this study we searched for mutations in a cohort of 38 patients and investigated the relationship between mutation type and allele-specific transcription from the wild-type versus mutant alleles. Quantification of relative mRNA transcript numbers was done by Pyrosequencing, a novel real-time sequencing method whose signals can be quantified very accurately. We identified 21 novel mutations comprising various mutation types. Pyrosequencing detected a definite relationship between allelic NF1 transcript imbalance due to NMD and mutation type in 24 of 29 patients who all carried frame-shift or nonsense mutations. NMD was absent in 5 patients with missense and silent mutations, as well as in 4 patients with splice-site mutations that did not disrupt the reading frame. Pyrosequencing was capable of detecting NMD even when the effects were only moderate. Diagnostic laboratories could thus exploit this effect for rapid prescreening for NF1 mutations as more than 60% of the mutations in this gene disrupt the reading frame and are prone to NMD.

Validation of the VE1 Immunostain for the BRAF V600E Mutation in Melanoma

PubMed Central

Pearlstein, Michelle V.; Zedek, Daniel C.; Ollila, David W.; Treece, Amanda; Gulley, Margaret L.; Groben, Pamela A.; Thomas, Nancy E.

2014-01-01

BACKGROUND BRAF mutation status, and therefore eligibility for BRAF inhibitors, is currently determined by sequencing methods. We assessed the validity of VE1, a monoclonal antibody against the BRAF V600E mutant protein, in the detection of mutant BRAF V600E melanomas as classified by DNA pyrosequencing. METHODS The cases were 76 metastatic melanoma patients with only one known primary melanoma who had had BRAF codon 600 pyrosequencing of either their primary (n=19), metastatic (n=57) melanoma, or both (n=17). All melanomas (n=93) were immunostained with the BRAF VE1 antibody using a red detection system. The staining intensity of these specimens was scored from 0 – 3+ by a dermatopathologist. Scores of 0 and 1+ were considered as negative staining while scores of 2+ and 3+ were considered positive. RESULTS The VE1 antibody demonstrated a sensitivity of 85% and a specificity of 100% as compared to DNA pyrosequencing results. There was 100% concordance between VE1 immunostaining of primary and metastatic melanomas from the same patient. V600K, V600Q, and V600R BRAF melanomas did not positively stain with VE1. CONCLUSIONS This hospital-based study finds high sensitivity and specificity for the BRAF VE1 immunostain in comparison to pyrosequencing in detection of BRAF V600E in melanomas. PMID:24917033

Partitioning of ocean and land uptake of CO2 as inferred by delta C-13 measurements from the NOAA Climate Monitoring and Diagnostics Laboratory Global Air Sampling Network

NASA Technical Reports Server (NTRS)

Ciais, Philippe; Tans, Pieter P.; White, James W. C.; Trolier, Michael; Francey, Roger J.; Berry, Joe A.; Randall, David R.; Sellers, Piers J.; Collatz, James G.; Schimel, David S.

1995-01-01

Using delta C-13 measurements in atmospheric CO2 from a cooperative global air sampling network, we determined the partitioning of the net uptake of CO2 between ocean and land as a function of latitude and time. The majority of delta C-13 measurements were made at the Institute of Arctic and Alpine Research (INSTAAR) of the University of Colorado. We perform an inverse deconvolution of both CO2 and delta C-13 observations, using a two-dimensional model of atmospheric transport. Also, the discrimination against C-13 by plant photosynthesis, as a function of latitude and time, is calculated from global runs of the simple biosphere (SiB) model. Uncertainty due to the longitudinal structure of the data, which is not represented by the model, is studied through a bootstrap analysis by adding and omitting measurement sites. The resulting error estimates for our inferred sources and sinks are of the order of 1 GTC (1 GTC = 10(exp 15) gC). Such error bars do not reflect potential systematic errors arising from our estimates of the isotopic disequilibria between the atmosphere and the oceans and biosphere, which are estimated in a separate sensitivity analysis. With respect to global totals for 1992 we found that 3.2 GTC of carbon dissolved into the ocean and that 1.5 GTC were sequestered by land ecosystems. Northern hemisphere ocean gyres north of 15 deg N absorbed 2.7 GTC. The equatorial oceans between 10 deg S and 10 deg N were a net source to the atmosphere of 0.9 GTC. We obtained a sink of 1.6 GTC in southern ocean gyres south of 20 deg S, although the deconvolution is poorly constrained by sparse data coverage at high southern latitudes. The seasonal uptake of CO2 in the northern gyres appears to be correlated with a bloom of phytoplankton in surface waters. On land, northern temperate and boreal ecosystems between 35 deg N and 65 deg N were found to be a major sink of CO2 in 1992, as large as 3.5 GTC. Northern tropical ecosystems (equator-30 deg N) appear to be a net source to the source to the atmosphere of 2 GTC which could reflect biomass burning. A small sink, 0.3 GTC, was inferred for southern tropical ecosystems (30 deg S-equator).

In situ Detection of Microbial Life in the Deep Biosphere in Igneous Ocean Crust.

PubMed

Salas, Everett C; Bhartia, Rohit; Anderson, Louise; Hug, William F; Reid, Ray D; Iturrino, Gerardo; Edwards, Katrina J

2015-01-01

The deep biosphere is a major frontier to science. Recent studies have shown the presence and activity of cells in deep marine sediments and in the continental deep biosphere. Volcanic lavas in the deep ocean subsurface, through which substantial fluid flow occurs, present another potentially massive deep biosphere. We present results from the deployment of a novel in situ logging tool designed to detect microbial life harbored in a deep, native, borehole environment within igneous oceanic crust, using deep ultraviolet native fluorescence spectroscopy. Results demonstrate the predominance of microbial-like signatures within the borehole environment, with densities in the range of 10(5) cells/mL. Based on transport and flux models, we estimate that such a concentration of microbial cells could not be supported by transport through the crust, suggesting in situ growth of these communities.

Understanding local community's values, worldviews and perceptions in the Galloway and Southern Ayrshire Biosphere Reserve, Scotland.

PubMed

Hernes, Maya I; Metzger, Marc J

2017-01-15

Biosphere reserves have been studied around the world, but methods to elicit community's values, worldviews and perceptions are missing. A greater understanding of these can help avoid tension and improve successful management. This paper used a mixed-methods survey to elicit local community's environmental values, ecological world views and perceptions of the Galloway and Southern Ayrshire Biosphere Reserve (GSABR). Over three weeks, forty participants from three communities of the GSABR responded to a semi-structured mixed-methods survey. The survey revealed that residents of the GSABR greatly value wildlife and beauty of nature, and that the majority of the respondents showed concern for the environment from an ecocentric worldview. Results also revealed that the most influential tested socio-demographic characteristic affecting people's relationship to their environment is their professional affiliation. Tourism and recreation were seen as major benefits of the recent biosphere designation. Results did highlight contrasting benefits from the designation for different stakeholder groups, which could potentially lead to tensions and should be considered in the reserve management. Given the community's supportive world views and perceptions, greater participation in the biosphere's management in likely to be welcomed and should be used to avoid or mediate any conflicts. The mixed-method survey developed for this study, proved successful in eliciting these themes in the GSABR. We recommend other biosphere reserves replicate this research, to gain better understanding of local communities and increase their support and participation in reserve management. Copyright © 2016 Elsevier Ltd. All rights reserved.

An Appropriate Cutoff Value for Determining the Colonization of Helicobacter pylori by the Pyrosequencing Method: Comparison with Conventional Methods.

PubMed

Kim, Jaeyeon; Kim, Nayoung; Jo, Hyun Jin; Park, Ji Hyun; Nam, Ryoung Hee; Seok, Yeong-Jae; Kim, Yeon-Ran; Kim, Joo Sung; Kim, Jung Mogg; Kim, Jung Min; Lee, Dong Ho; Jung, Hyun Chae

2015-10-01

Sequencing of 16S ribosomal RNA (rRNA) gene has improved the characterization of microbial communities. It enabled the detection of low abundance gastric Helicobacter pylori sequences even in subjects that were found to be H. pylori negative with conventional methods. The objective of this study was to obtain a cutoff value for H. pylori colonization in gastric mucosa samples by pyrosequencing method. Gastric mucosal biopsies were taken from 63 subjects whose H. pylori status was determined by a combination of serology, rapid urease test, culture, and histology. Microbial DNA from mucosal samples was amplified by PCR using universal bacterial primers. 16S rDNA amplicons were pyrosequenced. ROC curve analysis was performed to determine the cutoff value for H. pylori colonization by pyrosequencing. In addition, temporal changes in the stomach microbiota were observed in eight initially H. pylori-positive and eight H. pylori-negative subjects at a single time point 1-8 years later. Of the 63 subjects, the presence of H. pylori sequences was detected in all (28/28) conventionally H. pylori-positive samples and in 60% (21/35) of H. pylori-negative samples. The average percent of H. pylori reads in each sample was 0.67 ± 1.09% in the H. pylori-negative group. Cutoff value for clinically positive H. pylori status was approximately 1.22% based on ROC curve analysis (AUC = 0.957; p < .001). Helicobacter pylori was successfully eradicated in five of seven treated H. pylori-positive subjects (71.4%), and the percentage of H. pylori reads in these five subjects dropped from 1.3-95.18% to 0-0.16% after eradication. These results suggest that the cutoff value of H. pylori sequence percentage for H. pylori colonization by pyrosequencing could be set at approximately 1%. It might be helpful to analyze gastric microbiota related to H. pylori sequence status. © 2015 John Wiley & Sons Ltd.

Use of pyrosequencing and denaturing gradient gel electrophoresis to examine the effects of probiotics and essential oil blends on digestive microflora in broilers under mixed Eimeria infection.

PubMed

Hume, Michael E; Barbosa, Nei A; Dowd, Scot E; Sakomura, Nilva K; Nalian, Armen G; Martynova-Van Kley, Alexandra; Oviedo-Rondón, Edgar O

2011-11-01

A protective digestive microflora helps prevent and reduce broiler infection and colonization by enteropathogens. In the current experiment, broilers fed diets supplemented with probiotics and essential oil (EO) blends were infected with a standard mixed Eimeria spp. to determine effects of performance enhancers on ileal and cecal microbial communities (MCs). Eight treatment groups included four controls (uninfected-unmedicated [UU], unmedicated-infected, the antibiotic BMD plus the ionophore Coban as positive control, and the ionophore as negative control), and four treatments (probiotics BC-30 and Calsporin; and EO, Crina Poultry Plus, and Crina PoultryAF). Day-old broilers were raised to 14 days in floor pens on used litter and then were moved to Petersime batteries and inoculated at 15 days with mixed Eimeria spp. Ileal and cecal samples were collected at 14 days and 7 days postinfection. Digesta DNA was subjected to pyrosequencing for sequencing of individual cecal bacteria and denaturing gradient gel electrophoresis (DGGE) for determination of changes in ileal and cecal MC according to percentage similarity coefficient (%SC). Pyrosequencing is very sensitive detecting shifts in individual bacterial sequences, whereas DGGE is able to detect gross shifts in entire MC. These combined techniques offer versatility toward identifying feed additive and mild Eimeria infection modulation of broiler MC. Pyrosequencing detected 147 bacterial species sequences. Additionally, pyrosequencing revealed the presence of relatively low levels of the potential human enteropathogens Campylobacter sp. and four Shigella spp. as well as the potential poultry pathogen Clostridiun perfringens. Pre- and postinfection changes in ileal (56%SC) and cecal (78.5%SC) DGGE profiles resulted from the coccidia infection and with increased broiler age. Probiotics and EO changed MC from those seen in UU ilea and ceca. Results potentially reflect the performance enhancement above expectations in comparison to broilers not given the probiotics or the specific EO blends as feed supplements.

454 next generation-sequencing outperforms allele-specific PCR, Sanger sequencing, and pyrosequencing for routine KRAS mutation analysis of formalin-fixed, paraffin-embedded samples

PubMed Central

Altimari, Annalisa; de Biase, Dario; De Maglio, Giovanna; Gruppioni, Elisa; Capizzi, Elisa; Degiovanni, Alessio; D’Errico, Antonia; Pession, Annalisa; Pizzolitto, Stefano; Fiorentino, Michelangelo; Tallini, Giovanni

2013-01-01

Detection of KRAS mutations in archival pathology samples is critical for therapeutic appropriateness of anti-EGFR monoclonal antibodies in colorectal cancer. We compared the sensitivity, specificity, and accuracy of Sanger sequencing, ARMS-Scorpion (TheraScreen®) real-time polymerase chain reaction (PCR), pyrosequencing, chip array hybridization, and 454 next-generation sequencing to assess KRAS codon 12 and 13 mutations in 60 nonconsecutive selected cases of colorectal cancer. Twenty of the 60 cases were detected as wild-type KRAS by all methods with 100% specificity. Among the 40 mutated cases, 13 were discrepant with at least one method. The sensitivity was 85%, 90%, 93%, and 92%, and the accuracy was 90%, 93%, 95%, and 95% for Sanger sequencing, TheraScreen real-time PCR, pyrosequencing, and chip array hybridization, respectively. The main limitation of Sanger sequencing was its low analytical sensitivity, whereas TheraScreen real-time PCR, pyrosequencing, and chip array hybridization showed higher sensitivity but suffered from the limitations of predesigned assays. Concordance between the methods was k = 0.79 for Sanger sequencing and k > 0.85 for the other techniques. Tumor cell enrichment correlated significantly with the abundance of KRAS-mutated deoxyribonucleic acid (DNA), evaluated as ΔCt for TheraScreen real-time PCR (P = 0.03), percentage of mutation for pyrosequencing (P = 0.001), ratio for chip array hybridization (P = 0.003), and percentage of mutation for 454 next-generation sequencing (P = 0.004). Also, 454 next-generation sequencing showed the best cross correlation for quantification of mutation abundance compared with all the other methods (P < 0.001). Our comparison showed the superiority of next-generation sequencing over the other techniques in terms of sensitivity and specificity. Next-generation sequencing will replace Sanger sequencing as the reference technique for diagnostic detection of KRAS mutation in archival tumor tissues. PMID:23950653

Rumen bacterial community evaluated by 454 pyrosequencing and terminal restriction fragment length polymorphism analyses in dairy sheep fed marine algae.

PubMed

Castro-Carrera, T; Toral, P G; Frutos, P; McEwan, N R; Hervás, G; Abecia, L; Pinloche, E; Girdwood, S E; Belenguer, A

2014-03-01

Developing novel strategies to increase the content of bioactive unsaturated fatty acids (FA) in ruminant-derived products requires a deeper understanding of rumen biohydrogenation and bacteria involved in this process. Although high-throughput pyrosequencing may allow for a great coverage of bacterial diversity, it has hardly been used to investigate the microbiology of ruminal FA metabolism. In this experiment, 454 pyrosequencing and a molecular fingerprinting technique (terminal restriction fragment length polymorphism; T-RFLP) were used concurrently to assess the effect of diet supplementation with marine algae (MA) on the rumen bacterial community of dairy sheep. Eleven lactating ewes were divided in 2 lots and offered a total mixed ration based on alfalfa hay and concentrate (40:60), supplemented with 0 (control) or 8 (MA) g of MA/kg of dry matter. After 54 d on treatments, animals were slaughtered and samples of rumen content and fluid were collected separately for microbial analysis. Pyrosequencing yielded a greater coverage of bacterial diversity than T-RFLP and allowed the identification of low abundant populations. Conversely, both molecular approaches pointed to similar conclusions and showed that relevant changes due to MA addition were observed within the major ruminal phyla, namely Bacteroidetes, Firmicutes, and Proteobacteria. Decreases in the abundance of unclassified Bacteroidales, Porphyromonadaceae, and Ruminococcaceae and increases in as-yet uncultured species of the family Succinivibrionaceae, might be related to a potential role of these groups in different pathways of rumen FA metabolism. Diet supplementation with MA, however, had no effect on the relative abundance of Butyrivibrio and Pseudobutyrivibrio genera. In addition, results from both 454 pyrosequencing and T-RFLP indicate that the effect of MA was rather consistent in rumen content or fluid samples, despite inherent differences between these fractions in their bacterial composition. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Geospatial assessment and monitoring of historical forest cover changes (1920-2012) in Nilgiri Biosphere Reserve, Western Ghats, India.

PubMed

Satish, K V; Saranya, K R L; Reddy, C Sudhakar; Krishna, P Hari; Jha, C S; Rao, P V V Prasada

2014-12-01

Deforestation in the biosphere reserves, which are key Protected Areas has negative impacts on biodiversity, climate, carbon fluxes and livelihoods. Comprehensive study of deforestation in biosphere reserves is required to assess the impact of the management effectiveness. This article assesses the changes in forest cover in various zones and protected areas of Nilgiri Biosphere Reserve, the first declared biosphere reserve in India which forms part of Western Ghats-a global biodiversity hotspot. In this study, we have mapped the forests from earliest available topographical maps and multi-temporal satellite data spanning from 1920's to 2012 period. Mapping of spatial extent of forest cover, vegetation types and land cover was carried out using visual interpretation technique. A grid cell of 1 km × 1 km was generated for time series change analysis to understand the patterns in spatial distribution of forest cover (1920-1973-1989-1999-2006-2012). The total forest area of biosphere reserve was found to be 5,806.5 km(2) (93.8 % of total geographical area) in 1920. Overall loss of forest cover was estimated as 1,423.6 km(2) (24.5 % of the total forest) with reference to 1920. Among the six Protected Areas, annual deforestation rate of >0.5 was found in Wayanad wildlife sanctuary during 1920-1973. The deforestation in Nilgiri Biosphere Reserve is mainly attributed to conversion of forests to plantations and agriculture along with submergence due to construction of dams during 1920 to 1989. Grid wise analysis indicates that 851 grids have undergone large-scale negative changes of >75 ha of forest loss during 1920-1973 while, only 15 grids have shown >75 ha loss during 1973-1989. Annual net rate of deforestation for the period of 1920 to 1973 was calculated as 0.5 followed by 0.1 for 1973 to 1989. Our analysis shows that there was large-scale deforestation before the declaration of area as biosphere reserve in 1986; however, the deforestation has drastically reduced after the declaration due to high degree of protection, thus indicating the secure future of reserve in the long term under the current forest management practices. The present work will stand as the most up-to-date assessment on the forest cover of the Nilgiri Biosphere Reserve with immediate applications in monitoring and management of forest biodiversity.

A biosphere modeling methodology for dose assessments of the potential Yucca Mountain deep geological high level radioactive waste repository.

PubMed

Watkins, B M; Smith, G M; Little, R H; Kessler, J

1999-04-01

Recent developments in performance standards for proposed high level radioactive waste disposal at Yucca Mountain suggest that health risk or dose rate limits will likely be part of future standards. Approaches to the development of biosphere modeling and dose assessments for Yucca Mountain have been relatively lacking in previous performance assessments due to the absence of such a requirement. This paper describes a practical methodology used to develop a biosphere model appropriate for calculating doses from use of well water by hypothetical individuals due to discharges of contaminated groundwater into a deep well. The biosphere model methodology, developed in parallel with the BIOMOVS II international study, allows a transparent recording of the decisions at each step, from the specification of the biosphere assessment context through to model development and analysis of results. A list of features, events, and processes relevant to Yucca Mountain was recorded and an interaction matrix developed to help identify relationships between them. Special consideration was given to critical/potential exposure group issues and approaches. The conceptual model of the biosphere system was then developed, based on the interaction matrix, to show how radionuclides migrate and accumulate in the biosphere media and result in potential exposure pathways. A mathematical dose assessment model was specified using the flexible AMBER software application, which allows users to construct their own compartment models. The starting point for the biosphere calculations was a unit flux of each radionuclide from the groundwater in the geosphere into the drinking water in the well. For each of the 26 radionuclides considered, the most significant exposure pathways for hypothetical individuals were identified. For 14 of the radionuclides, the primary exposure pathways were identified as consumption of various crops and animal products following assumed agricultural use of the contaminated water derived from the deep well. Inhalation of dust (11 radionuclides) and external irradiation (1 radionuclide) were also identified as significant exposure modes. Contribution to the total flux to dose conversion factor from the drinking water pathway for each radionuclide was also assessed and for most radionuclides was found to be less than 10% of the total flux to dose conversion factor summed across all pathways. Some of the uncertainties related to the results were considered. The biosphere modeling results have been applied within an EPRI Total Systems Performance Assessment of Yucca Mountain. Conclusions and recommendations for future performance assessments are provided.

Earth applications of closed ecological systems: Relevance to the development of sustainability in our global biosphere

NASA Astrophysics Data System (ADS)

Nelson, M.; Allen, J.; Ailing, A.; Dempster, W. F.; Silverstone, S.

The parallels between the challenges facing bioregenerative life support in artificial closed ecological systems and those in our global biosphere are striking. At the scale of the current global technosphere and expanding human population, it is increasingly obvious that the biosphere can no longer safely buffer and absorb technogenic and anthropogenic pollutants. The loss of biodiversity, reliance on non-renewable natural resources, and conversion of once wild ecosystems for human use with attendant desertification/soil erosion, has led to a shift of consciousness and the widespread call for sustainability of human activities. For researchers working on bioregenerative life support in closed systems, the small volumes and faster cycling times than in the Earth's biosphere make it starkly clear that systems must be designed to ensure renewal of water and atmosphere, nutrient recycling, production of healthy food, and safe environmental methods of maintaining technical systems. The development of technical systems that can be fully integrated and supportive of living systems is a harbinger of new perspectives as well as technologies in the global environment. In addition, closed system bioregenerative life support offers opportunities for public education and consciousness changing of how to live with our global biosphere.

A Bioluminometric Method of DNA Sequencing

NASA Technical Reports Server (NTRS)

Ronaghi, Mostafa; Pourmand, Nader; Stolc, Viktor; Arnold, Jim (Technical Monitor)

2001-01-01

Pyrosequencing is a bioluminometric single-tube DNA sequencing method that takes advantage of co-operativity between four enzymes to monitor DNA synthesis. In this sequencing-by-synthesis method, a cascade of enzymatic reactions yields detectable light, which is proportional to incorporated nucleotides. Pyrosequencing has the advantages of accuracy, flexibility and parallel processing. It can be easily automated. Furthermore, the technique dispenses with the need for labeled primers, labeled nucleotides and gel-electrophoresis. In this chapter, the use of this technique for different applications is discussed.

The Role of Biosphere Reserves in Environmental Education and Training = Le Role des reserves de la biosphere dans l'education et la formation environnementales. Report of the Unesco/MAB Symposium Held During the Unesco/UNEP International Congress on Environmental Education and Training (Moscow, USSR, August 17-21, 1987). Report 20.

ERIC Educational Resources Information Center

Francis, George, Ed.

Environmental education and training have been key elements of Unesco's Program on Man and the Biosphere (MAB) since its inception in 1971. The MAB Program is an intergovernmental program of research, training, demonstration and distribution of information, aimed at providing the scientific background and the trained personnel to deal with…

Ecology and exploration of the rare biosphere.

PubMed

Lynch, Michael D J; Neufeld, Josh D

2015-04-01

The profound influence of microorganisms on human life and global biogeochemical cycles underlines the value of studying the biogeography of microorganisms, exploring microbial genomes and expanding our understanding of most microbial species on Earth: that is, those present at low relative abundance. The detection and subsequent analysis of low-abundance microbial populations—the 'rare biosphere'—have demonstrated the persistence, population dynamics, dispersion and predation of these microbial species. We discuss the ecology of rare microbial populations, and highlight molecular and computational methods for targeting taxonomic 'blind spots' within the rare biosphere of complex microbial communities.

On the edge of a deep biosphere: Real animals in extreme environments

NASA Astrophysics Data System (ADS)

Childress, James J.; Fisher, Charles F.; Felbeck, Horst; Girguis, Peter

This paper considers the possibility of animals living in a subsurface environment on the global mid-ocean ridge system. It considers the possible environments and looks at the possibilities of animal inhabitants of the subsurface biosphere based on adaptations of animals to other extreme habitats. We conclude that there are known bridging inhabitants of the subsurface biosphere, that part-time inhabitants are extremely likely, and that there could be full-time inhabitants if conditions are stable within the tolerance limits of metazoans for time periods of months.

Victor Kovda, Soil Science and Biosphere

NASA Astrophysics Data System (ADS)

Kovda, I.

2012-04-01

Victor Kovda (1904-1991) was one of the most famous soil scientists at the national and international soil science community. He published more than 500 scientific works including about 400 papers, 17 collective monographs, 30 personal monographs, and more than 200 interviews and popular papers describing the role of soils not only for food production, but for the functioning of the biosphere. Victor Kovda was a talented organizer, who founded the new Institute of Soil Science and Agrochemistry (known at the present time as the Institute of physico-chemical and biological problems of soil science in Pushchino, Russia). During six years from 1959 to 1964 he was the head of Science Department in UNESCO, where he initiated a set of international projects (ex. Soil World Map of FAO-UNESCO, Source-book on irrigation and drainage). He continued his international activity after UNESCO as a President of the International Soil Science Society (1968-1974), organizer of the X international Soil Science Congress in Moscow (1974), president of SCOPE (1973-1976), working for ICSU. The last three decades of his national and international activities Victor Kovda initiated and was strongly involved in the popularization of biosphere role and functions of soils and soil cover. The start point for this activity was his special talk "Biosphere and man" presented during the intergovernmental conference in the framework of the international program "Man and Biosphere" organized by UNESCO in 1968 in Paris. The next key presentation "Soil as a component of biosphere" Victor Kovda gave as a plenary lecture during the X International congress of soil scientists. This presentation determined the focus of soil science for the next decades: at least Russian soil science became oriented towards the investigation of biosphere functions and role of soils. Soils science was accepted not only for agriculture and food production, but also as a fundamental science with a large environmental application.

The Search for Sustainable Subsurface Habitats on Mars, and the Sampling of Impact Ejecta

NASA Astrophysics Data System (ADS)

Ivarsson, Magnus; Lindgren, Paula

2010-07-01

On Earth, the deep subsurface biosphere of both the oceanic and the continental crust is well known for surviving harsh conditions and environments characterized by high temperatures, high pressures, extreme pHs, and the absence of sunlight. The microorganisms of the terrestrial deep biosphere have an excellent capacity for adapting to changing geochemistry, as the alteration of the crust proceeds and the conditions of their habitats slowly change. Despite an almost complete isolation from surface conditions and the surface biosphere, the deep biosphere of the crustal rocks has endured over geologic time. This indicates that the deep biosphere is a self-sufficient system, independent of the global events that occur at the surface, such as impacts, glaciations, sea level fluctuations, and climate changes. With our sustainable terrestrial subsurface biosphere in mind, the subsurface on Mars has often been suggested as the most plausible place to search for fossil Martian life, or even present Martian life. Since the Martian surface is more or less sterile, subsurface settings are the only place on Mars where life could have been sustained over geologic time. To detect a deep biosphere in the Martian basement, drilling is a requirement. However, near future Mars sample return missions are limited by the mission's payload, which excludes heavy drilling equipment and restrict the missions to only dig the topmost meter of the Martian soil. Therefore, the sampling and analysis of Martian impact ejecta has been suggested as a way of accessing the deeper Martian subsurface without using heavy drilling equipment. Impact cratering is a natural geological process capable of excavating and exposing large amounts of rock material from great depths up to the surface. Several studies of terrestrial impact deposits show the preservation of pre-impact biosignatures, such as fossilized organisms and chemical biological markers. Therefore, if the Martian subsurface contains a record of life, it is reasonable to assume that biosignatures derived from the Martian subsurface could also be preserved in the Martian impact ejecta.

Pilot Institute on Global Change on Trace Gases and the Biosphere, 1988

NASA Technical Reports Server (NTRS)

Eddy, J. A.; Moore, B.

1998-01-01

Table of Contents: Summary; Background; General Framework for a Series of Institutes on Global Change; The 1988 Pilot Institute on Global Changes: Trace Gases and the Biosphere; Budget; List of Acronyms; and Attachments.

Closed ecological systems: From test tubes to Earth's biosphere

NASA Technical Reports Server (NTRS)

Frye, Robert J.; Mignon, George

1992-01-01

Artificially constructed closed ecological systems (CES) have been researched both experimentally and theoretically for over 25 years. The size of these systems have varied from less than one liter to many thousands of cubic meters in volume. The diversity of the included components has a similarly wide range from purely aquatic systems to soil based systems that incorporate many aspects of Earth's biosphere. While much has been learned about the functioning of these closed systems, much remains to be learned. In this paper, we compare and contrast the behavior of closed ecological systems of widely different sizes through an analysis of their atmospheric composition. In addition, we will compare the performance of relatively small CES with the behavior of Earth's biosphere. We address the applicability of small CES as replicable analogs for planetary biospheres and discuss the use of small CES as an experimental milieu for an examination of the evolution of extra-terrestrial colonies.

In situ Detection of Microbial Life in the Deep Biosphere in Igneous Ocean Crust

PubMed Central

Salas, Everett C.; Bhartia, Rohit; Anderson, Louise; Hug, William F.; Reid, Ray D.; Iturrino, Gerardo; Edwards, Katrina J.

2015-01-01

The deep biosphere is a major frontier to science. Recent studies have shown the presence and activity of cells in deep marine sediments and in the continental deep biosphere. Volcanic lavas in the deep ocean subsurface, through which substantial fluid flow occurs, present another potentially massive deep biosphere. We present results from the deployment of a novel in situ logging tool designed to detect microbial life harbored in a deep, native, borehole environment within igneous oceanic crust, using deep ultraviolet native fluorescence spectroscopy. Results demonstrate the predominance of microbial-like signatures within the borehole environment, with densities in the range of 105 cells/mL. Based on transport and flux models, we estimate that such a concentration of microbial cells could not be supported by transport through the crust, suggesting in situ growth of these communities. PMID:26617595

Gene expression in the deep biosphere.

PubMed

Orsi, William D; Edgcomb, Virginia P; Christman, Glenn D; Biddle, Jennifer F

2013-07-11

Scientific ocean drilling has revealed a deep biosphere of widespread microbial life in sub-seafloor sediment. Microbial metabolism in the marine subsurface probably has an important role in global biogeochemical cycles, but deep biosphere activities are not well understood. Here we describe and analyse the first sub-seafloor metatranscriptomes from anaerobic Peru Margin sediment up to 159 metres below the sea floor, represented by over 1 billion complementary DNA (cDNA) sequence reads. Anaerobic metabolism of amino acids, carbohydrates and lipids seem to be the dominant metabolic processes, and profiles of dissimilatory sulfite reductase (dsr) transcripts are consistent with pore-water sulphate concentration profiles. Moreover, transcripts involved in cell division increase as a function of microbial cell concentration, indicating that increases in sub-seafloor microbial abundance are a function of cell division across all three domains of life. These data support calculations and models of sub-seafloor microbial metabolism and represent the first holistic picture of deep biosphere activities.

Community Assembly Processes of the Microbial Rare Biosphere.

PubMed

Jia, Xiu; Dini-Andreote, Francisco; Falcão Salles, Joana

2018-03-14

Our planet teems with microorganisms that often present a skewed abundance distribution in a local community, with relatively few dominant species coexisting alongside a high number of rare species. Recent studies have demonstrated that these rare taxa serve as limitless reservoirs of genetic diversity, and perform disproportionate types of functions despite their low abundances. However, relatively little is known about the mechanisms controlling rarity and the processes promoting the development of the rare biosphere. Here, we propose the use of multivariate cut-offs to estimate rare species and phylogenetic null models applied to predefined rare taxa to disentangle the relative influences of ecoevolutionary processes mediating the assembly of the rare biosphere. Importantly, the identification of the factors controlling rare species assemblages is critical for understanding the types of rarity, how the rare biosphere is established, and how rare microorganisms fluctuate over spatiotemporal scales, thus enabling prospective predictions of ecosystem responses. Copyright © 2018 Elsevier Ltd. All rights reserved.

Picoliter DNA Sequencing Chemistry on an Electrowetting-based Digital Microfluidic Platform

PubMed Central

Ferguson Welch, Erin R.; Lin, Yan-You; Madison, Andrew; Fair, R.B.

2011-01-01

The results of investigations into performing DNA sequencing chemistry on a picoliter-scale electrowetting digital microfluidic platform are reported. Pyrosequencing utilizes pyrophosphate produced during nucleotide base addition to initiate a process ending with detection through a chemiluminescence reaction using firefly luciferase. The intensity of light produced during the reaction can be quantified to determine the number of bases added to the DNA strand. The logic-based control and discrete fluid droplets of a digital microfluidic device lend themselves well to the pyrosequencing process. Bead-bound DNA is magnetically held in a single location, and wash or reagent droplets added or split from it to circumvent product dilution. Here we discuss the dispensing, control, and magnetic manipulation of the paramagnetic beads used to hold target DNA. We also demonstrate and characterize the picoliter-scale reaction of luciferase with adenosine triphosphate to represent the detection steps of pyrosequencing and all necessary alterations for working on this scale. PMID:21298802

Rapid Molecular Identification of Human Taeniid Cestodes by Pyrosequencing Approach

PubMed Central

Thanchomnang, Tongjit; Tantrawatpan, Chairat; Intapan, Pewpan M.; Sanpool, Oranuch; Janwan, Penchom; Lulitanond, Viraphong; Tourtip, Somjintana; Yamasaki, Hiroshi; Maleewong, Wanchai

2014-01-01

Taenia saginata, T. solium, and T. asiatica are causative agents of taeniasis in humans. The difficulty of morphological identification of human taeniids can lead to misdiagnosis or confusion. To overcome this problem, several molecular methods have been developed, but use of these tends to be time-consuming. Here, a rapid and high-throughput pyrosequencing approach was developed for the identification of three human taeniids originating from various countries. Primers targeting the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of the three Taenia species were designed. Variations in a 26-nucleotide target region were used for identification. The reproducibility and accuracy of the pyrosequencing technology was confirmed by Sanger sequencing. This technique will be a valuable tool to distinguish between sympatric human taeniids that occur in Thailand, Asia and Pacific countries. This method could potentially be used for the molecular identification of the taeniid species that might be associated with suspicious cysts and lesions, or cyst residues in humans or livestock at the slaughterhouse. PMID:24945530

A comparative radiological assessment of five European biosphere systems in the context of potential contamination of well water from the hypothetical disposal of radioactive waste.

PubMed

Olyslaegers, G; Zeevaert, T; Pinedo, P; Simon, I; Pröhl, G; Kowe, R; Chen, Q; Mobbs, S; Bergström, U; Hallberg, B; Katona, T; Eged, K; Kanyar, B

2005-12-01

In the framework of the BioMoSA project for the development of biosphere assessment models for radioactive waste disposal the Reference Biosphere Methodology developed in the IAEA programme BIOMASS was applied to five locations, situated in different European countries. Specific biosphere models were applied to assess the hypothetical contamination of a range of agricultural and environmental pathways and the dose to individuals, following contamination of well water. The results of these site-specific models developed by the different BioMoSA partners, and the individual normalised dose to the exposure groups were compared against each other. Ingestion of drinking water, fruit and vegetables were found to be among the most important pathways for almost all radionuclides. Stochastic calculations revealed that consumption habits, transfer factors, irrigation rates and distribution coefficients (Kd(s)) were the most important parameters that influence the end results. Variations in the confidence intervals were found to be higher for sorbing elements (e.g. (36)Cl, (237)Np, (99)Tc, (238)U, (129)I) than for mobile elements (e.g. (226)Ra, (79)Se, (135)Cs, (231)Pa, (239)Pu). The influence of daughter products, for which the distribution into the biosphere was calculated individually, was also shown to be important. This paper gives a brief overview of the deterministic and stochastic modelling results and the parameter sensitivity. A screening methodology was introduced to identify the most important pathways, simplify a generic biosphere tool and refine the existing models.

Pyrosequencing analysis of the gyrB gene to differentiate bacteria responsible for diarrheal diseases.

PubMed

Hou, X-L; Cao, Q-Y; Jia, H-Y; Chen, Z

2008-07-01

Pathogens causing acute diarrhea include a large variety of species from Enterobacteriaceae and Vibrionaceae. A method based on pyrosequencing was used here to differentiate bacteria commonly associated with diarrhea in China; the method is targeted to a partial amplicon of the gyrB gene, which encodes the B subunit of DNA gyrase. Twenty-eight specific polymorphic positions were identified from sequence alignment of a large sequence dataset and targeted using 17 sequencing primers. Of 95 isolates tested, belonging to 13 species within 7 genera, most could be identified to the species level; O157 type could be differentiated from other E. coli types; Salmonella enterica subsp. enterica could be identified at the serotype level; the genus Shigella, except for S. boydii and S. dysenteriae, could also be identified. All these isolates were also subjected to conventional sequencing of a relatively long ( approximately1.2 kb) region of gyrB DNA; these results confirmed those with pyrosequencing. Twenty-two fecal samples were surveyed, the results of which were concordant with culture-based bacterial identification, and the pathogen detection limit with simulated stool specimens was 10(4) CFU/ml. DNA from different pathogens was also mixed to simulate a case of multibacterial infection, and the generated signals correlated well with the mix ratio. In summary, the gyrB-based pyrosequencing approach proved to have significant reliability and discriminatory power for enteropathogenic bacterial identification and provided a fast and effective method for clinical diagnosis.

Carbon 13 exchanges between the atmosphere and biosphere

NASA Astrophysics Data System (ADS)

Fung, I.; Field, C. B.; Berry, J. A.; Thompson, M. V.; Randerson, J. T.; MalmströM, C. M.; Vitousek, P. M.; Collatz, G. James; Sellers, P. J.; Randall, D. A.; Denning, A. S.; Badeck, F.; John, J.

1997-12-01

We present a detailed investigation of the gross 12C and 13C exchanges between the atmosphere and biosphere and their influence on the δ13C variations in the atmosphere. The photosynthetic discrimination Δ against 13C is derived from a biophysical model coupled to a general circulation model [Sellers et al., 1996a], where stomatal conductance and carbon assimilation are determined simultaneously with the ambient climate. The δ13C of the respired carbon is calculated by a biogeochemical model [Potter et al., 1993; Randerson et al., 1996] as the sum of the contributions from compartments with varying ages. The global flux-weighted mean photosynthetic discrimination is 12-16‰, which is lower than previous estimates. Factors that lower the discrimination are reduced stomatal conductance and C4 photosynthesis. The decreasing atmospheric δ13C causes an isotopic disequilibrium between the outgoing and incoming fluxes; the disequilibrium is ˜0.33‰ for 1988. The disequilibrium is higher than previous estimates because it accounts for the lifetime of trees and for the ages rather than turnover times of the biospheric pools. The atmospheric δ13C signature resulting from the biospheric fluxes is investigated using a three-dimensional atmospheric tracer model. The isotopic disequilibrium alone produces a hemispheric difference of ˜0.02‰ in atmospheric δ13C, comparable to the signal from a hypothetical carbon sink of 0.5 Gt C yr-1 into the midlatitude northern hemisphere biosphere. However, the rectifier effect, due to the seasonal covariation of CO2 fluxes and height of the atmospheric boundary layer, yields a background δ13C gradient of the opposite sign. These effects nearly cancel thus favoring a stronger net biospheric uptake than without the background CO2 gradient. Our analysis of the globally averaged carbon budget for the decade of the 1980s indicates that the biospheric uptake of fossil fuel CO2 is likely to be greater than the oceanic uptake; the relative proportions of the sinks cannot be uniquely determined using 12C and 13C alone. The land-ocean sink partitioning requires, in addition, information about the land use source, isotopic disequilibrium associated with gross oceanic exchanges, as well as the fractions of C3 and C4 vegetation involved in the biospheric uptake.

Exploring global carbon turnover and radiocarbon cycling in terrestrial biosphere models

NASA Astrophysics Data System (ADS)

Graven, H. D.; Warren, H.

2017-12-01

The uptake of carbon into terrestrial ecosystems through net primary productivity (NPP) and the turnover of that carbon through various pathways are the fundamental drivers of changing carbon stocks on land, in addition to human-induced and natural disturbances. Terrestrial biosphere models use different formulations for carbon uptake and release, resulting in a range of values in NPP of 40-70 PgC/yr and biomass turnover times of about 25-40 years for the preindustrial period in current-generation models from CMIP5. Biases in carbon uptake and turnover impact simulated carbon uptake and storage in the historical period and later in the century under changing climate and CO2 concentration, however evaluating global-scale NPP and carbon turnover is challenging. Scaling up of plot-scale measurements involves uncertainty due to the large heterogeneity across ecosystems and biomass types, some of which are not well-observed. We are developing the modelling of radiocarbon in terrestrial biosphere models, with a particular focus on decadal 14C dynamics after the nuclear weapons testing in the 1950s-60s, including the impact of carbon flux trends and variability on 14C cycling. We use an estimate of the total inventory of excess 14C in the biosphere constructed by Naegler and Levin (2009) using a 14C budget approach incorporating estimates of total 14C produced by the weapons tests and atmospheric and oceanic 14C observations. By simulating radiocarbon in simple biosphere box models using carbon fluxes from the CMIP5 models, we find that carbon turnover is too rapid in many of the simple models - the models appear to take up too much 14C and release it too quickly. Therefore many CMIP5 models may also simulate carbon turnover that is too rapid. A caveat is that the simple box models we use may not adequately represent carbon dynamics in the full-scale models. Explicit simulation of radiocarbon in terrestrial biosphere models would allow more robust evaluation of biosphere models and the investigation of climate-carbon cycle feedbacks on various timescales. Explicit simulation of radiocarbon and carbon-13 in terrestrial biosphere models of Earth System Models, as well as in ocean models, is recommended by CMIP6 and supported by CMIP6 protocols and forcing datasets.

Viral quasispecies inference from 454 pyrosequencing

PubMed Central

2013-01-01

Background Many potentially life-threatening infectious viruses are highly mutable in nature. Characterizing the fittest variants within a quasispecies from infected patients is expected to allow unprecedented opportunities to investigate the relationship between quasispecies diversity and disease epidemiology. The advent of next-generation sequencing technologies has allowed the study of virus diversity with high-throughput sequencing, although these methods come with higher rates of errors which can artificially increase diversity. Results Here we introduce a novel computational approach that incorporates base quality scores from next-generation sequencers for reconstructing viral genome sequences that simultaneously infers the number of variants within a quasispecies that are present. Comparisons on simulated and clinical data on dengue virus suggest that the novel approach provides a more accurate inference of the underlying number of variants within the quasispecies, which is vital for clinical efforts in mapping the within-host viral diversity. Sequence alignments generated by our approach are also found to exhibit lower rates of error. Conclusions The ability to infer the viral quasispecies colony that is present within a human host provides the potential for a more accurate classification of the viral phenotype. Understanding the genomics of viruses will be relevant not just to studying how to control or even eradicate these viral infectious diseases, but also in learning about the innate protection in the human host against the viruses. PMID:24308284

Tracking the ecological overshoot of the human economy

PubMed Central

Wackernagel, Mathis; Schulz, Niels B.; Deumling, Diana; Linares, Alejandro Callejas; Jenkins, Martin; Kapos, Valerie; Monfreda, Chad; Loh, Jonathan; Myers, Norman; Norgaard, Richard; Randers, Jørgen

2002-01-01

Sustainability requires living within the regenerative capacity of the biosphere. In an attempt to measure the extent to which humanity satisfies this requirement, we use existing data to translate human demand on the environment into the area required for the production of food and other goods, together with the absorption of wastes. Our accounts indicate that human demand may well have exceeded the biosphere's regenerative capacity since the 1980s. According to this preliminary and exploratory assessment, humanity's load corresponded to 70% of the capacity of the global biosphere in 1961, and grew to 120% in 1999. PMID:12089326

Earth applications of closed ecological systems: relevance to the development of sustainability in our global biosphere.

PubMed

Nelson, M; Allen, J; Alling, A; Dempster, W F; Silverstone, S

2003-01-01

The parallels between the challenges facing bioregenerative life support in artificial closed ecological systems and those in our global biosphere are striking. At the scale of the current global technosphere and expanding human population, it is increasingly obvious that the biosphere can no longer safely buffer and absorb technogenic and anthropogenic pollutants. The loss of biodiversity, reliance on non-renewable natural resources, and conversion of once wild ecosystems for human use with attendant desertification/soil erosion, has led to a shift of consciousness and the widespread call for sustainability of human activities. For researchers working on bioregenerative life support in closed systems, the small volumes and faster cycling times than in the Earth's biosphere make it starkly clear that systems must be designed to ensure renewal of water and atmosphere, nutrient recycling, production of healthy food, and safe environmental methods of maintaining technical systems. The development of technical systems that can be fully integrated and supportive of living systems is a harbinger of new perspectives as well as technologies in the global environment. In addition, closed system bioregenerative life support offers opportunities for public education and consciousness changing of how to live with our global biosphere. c2003 COSPAR. Published by Elsevier Science Ltd. All rights reserved.

Modeling the impact of climate change in Germany with biosphere models for long-term safety assessment of nuclear waste repositories.

PubMed

Staudt, C; Semiochkina, N; Kaiser, J C; Pröhl, G

2013-01-01

Biosphere models are used to evaluate the exposure of populations to radionuclides from a deep geological repository. Since the time frame for assessments of long-time disposal safety is 1 million years, potential future climate changes need to be accounted for. Potential future climate conditions were defined for northern Germany according to model results from the BIOCLIM project. Nine present day reference climate regions were defined to cover those future climate conditions. A biosphere model was developed according to the BIOMASS methodology of the IAEA and model parameters were adjusted to the conditions at the reference climate regions. The model includes exposure pathways common to those reference climate regions in a stylized biosphere and relevant to the exposure of a hypothetical self-sustaining population at the site of potential radionuclide contamination from a deep geological repository. The end points of the model are Biosphere Dose Conversion factors (BDCF) for a range of radionuclides and scenarios normalized for a constant radionuclide concentration in near-surface groundwater. Model results suggest an increased exposure of in dry climate regions with a high impact of drinking water consumption rates and the amount of irrigation water used for agriculture. Copyright © 2012 Elsevier Ltd. All rights reserved.

Characterization of microsatellite loci from two-spotted octopus Octopus bimaculatus Verrill 1883 from pyrosequencing reads

USGS Publications Warehouse

Domínguez-Contreras, J. F.; Munguía-Vega, A.; Ceballos-Vázquez, B. P.; Arellano-Martínez, M.; Culver, Melanie

2014-01-01

We characterized 22 novel microsatellite loci in the two-spotted octopus Octopus bimaculatus using 454 pyrosequencing reads. All loci were polymorphic and will be used in studies of marine connectivity aimed at increasing sustainability of the resource. The mean number alleles per locus was 13.09 (range 7–19) and observed heterozygosities ranged from 0.50 to 1.00. Four loci pairs were linked and three deviated from Hardy–Weinberg equilibrium. Eighteen and 12 loci were polymorphic in Octopus bimaculoides and Octopus hubbsorum, respectively.

Group dynamics challenges: Insights from Biosphere 2 experiments

NASA Astrophysics Data System (ADS)

Nelson, Mark; Gray, Kathelin; Allen, John P.

2015-07-01

Successfully managing group dynamics of small, physically isolated groups is vital for long duration space exploration/habitation and for terrestrial CELSS (Controlled Environmental Life Support System) facilities with human participants. Biosphere 2 had important differences and shares some key commonalities with both Antarctic and space environments. There were a multitude of stress factors during the first two year closure experiment as well as mitigating factors. A helpful tool used at Biosphere 2 was the work of W.R. Bion who identified two competing modalities of behavior in small groups. Task-oriented groups are governed by conscious acceptance of goals, reality-thinking in relation to time and resources, and intelligent management of challenges. The opposing unconscious mode, the "basic-assumption" ("group animal") group, manifests through Dependency/Kill the Leader, Fight/Flight and Pairing. These unconscious dynamics undermine and can defeat the task group's goal. The biospherians experienced some dynamics seen in other isolated teams: factions developing reflecting personal chemistry and disagreements on overall mission procedures. These conflicts were exacerbated by external power struggles which enlisted support of those inside. Nevertheless, the crew evolved a coherent, creative life style to deal with some of the deprivations of isolation. The experience of the first two year closure of Biosphere 2 vividly illustrates both vicissitudes and management of group dynamics. The crew overrode inevitable frictions to creatively manage both operational and research demands and opportunities of the facility, thus staying 'on task' in Bion's group dynamics terminology. The understanding that Biosphere 2 was their life support system may also have helped the mission to succeed. Insights from the Biosphere 2 experience can help space and remote missions cope successfully with the inherent challenges of small, isolated crews.

The Effects of Nitrogen Fertilization of a Corn Ecosystem's Oxidative Ratio and Its Carbon Cycle Implications

NASA Astrophysics Data System (ADS)

Gallagher, M. E.; Masiello, C. A.; Hockaday, W. C.; McSwiney, C. P.; Robertson, G. P.

2008-12-01

One of the most effective ways to estimate the size of carbon sinks in the terrestrial biosphere and oceans is through paired measurements of atmospheric CO2 and O2 concentrations (e.g. (Keeling et al. 1996)). Successful use of this technique requires knowledge of the oxidative ratio (OR) of the terrestrial biosphere (the ratio of moles of O2 released per moles of CO2 consumed in gas fluxes between the terrestrial biosphere and atmosphere.) Historically the terrestrial biosphere's OR has been assumed to be a constant, approximately 1.1 (e.g. Prentice et al. 2001). However, small shifts in the biosphere's OR values can lead to large variations in the calculated sizes of the terrestrial biosphere and ocean carbon sinks (Randerson et al. 2006). We have recently shown that it is possible to measure the OR of biomass to at least +/- 0.01 units (Masiello et al., 2008), and that there is significant natural variability in ecosystem OR. Ecosystem OR is impacted by human activities. In this presentation, we explore the effects of one major form of anthropogenic ecosystem alteration: nitrogen fertilization. We are measuring ecosystem OR in corn agricultural ecosystems under a range of nitrogen fertilization treatments at the Kellogg Biological Station- Long Term Ecological Research Site (KBS-LTER) in Michigan. We measure OR indirectly, through its relationship with organic carbon oxidation state (Cox) (Masiello et al. 2008). Here we present data showing the effects of varying corn ecosystem nitrogen fertilization rates (from 0 to 202 kg N/ha) on ecosystem OR and the implications it will have on apportionment calculations.

An improved land biosphere module for use in the DCESS Earth system model (version 1.1) with application to the last glacial termination

NASA Astrophysics Data System (ADS)

Eichinger, Roland; Shaffer, Gary; Albarrán, Nelson; Rojas, Maisa; Lambert, Fabrice

2017-09-01

Interactions between the land biosphere and the atmosphere play an important role for the Earth's carbon cycle and thus should be considered in studies of global carbon cycling and climate. Simple approaches are a useful first step in this direction but may not be applicable for certain climatic conditions. To improve the ability of the reduced-complexity Danish Center for Earth System Science (DCESS) Earth system model DCESS to address cold climate conditions, we reformulated the model's land biosphere module by extending it to include three dynamically varying vegetation zones as well as a permafrost component. The vegetation zones are formulated by emulating the behaviour of a complex land biosphere model. We show that with the new module, the size and timing of carbon exchanges between atmosphere and land are represented more realistically in cooling and warming experiments. In particular, we use the new module to address carbon cycling and climate change across the last glacial transition. Within the constraints provided by various proxy data records, we tune the DCESS model to a Last Glacial Maximum state and then conduct transient sensitivity experiments across the transition under the application of explicit transition functions for high-latitude ocean exchange, atmospheric dust, and the land ice sheet extent. We compare simulated time evolutions of global mean temperature, pCO2, atmospheric and oceanic carbon isotopes as well as ocean dissolved oxygen concentrations with proxy data records. In this way we estimate the importance of different processes across the transition with emphasis on the role of land biosphere variations and show that carbon outgassing from permafrost and uptake of carbon by the land biosphere broadly compensate for each other during the temperature rise of the early last deglaciation.

Towards understanding the variability in biospheric CO2 fluxes: using FTIR spectrometry and a chemical transport model to investigate the sources and sinks of carbonyl sulfide and its link to CO2

NASA Astrophysics Data System (ADS)

Wang, Yuting; Deutscher, Nicholas M.; Palm, Mathias; Warneke, Thorsten; Notholt, Justus; Baker, Ian; Berry, Joe; Suntharalingam, Parvadha; Jones, Nicholas; Mahieu, Emmanuel; Lejeune, Bernard; Hannigan, James; Conway, Stephanie; Mendonca, Joseph; Strong, Kimberly; Campbell, J. Elliott; Wolf, Adam; Kremser, Stefanie

2016-02-01

Understanding carbon dioxide (CO2) biospheric processes is of great importance because the terrestrial exchange drives the seasonal and interannual variability of CO2 in the atmosphere. Atmospheric inversions based on CO2 concentration measurements alone can only determine net biosphere fluxes, but not differentiate between photosynthesis (uptake) and respiration (production). Carbonyl sulfide (OCS) could provide an important additional constraint: it is also taken up by plants during photosynthesis but not emitted during respiration, and therefore is a potential means to differentiate between these processes. Solar absorption Fourier Transform InfraRed (FTIR) spectrometry allows for the retrievals of the atmospheric concentrations of both CO2 and OCS from measured solar absorption spectra. Here, we investigate co-located and quasi-simultaneous FTIR measurements of OCS and CO2 performed at five selected sites located in the Northern Hemisphere. These measurements are compared to simulations of OCS and CO2 using a chemical transport model (GEOS-Chem). The coupled biospheric fluxes of OCS and CO2 from the simple biosphere model (SiB) are used in the study. The CO2 simulation with SiB fluxes agrees with the measurements well, while the OCS simulation reproduced a weaker drawdown than FTIR measurements at selected sites, and a smaller latitudinal gradient in the Northern Hemisphere during growing season when comparing with HIPPO (HIAPER Pole-to-Pole Observations) data spanning both hemispheres. An offset in the timing of the seasonal cycle minimum between SiB simulation and measurements is also seen. Using OCS as a photosynthesis proxy can help to understand how the biospheric processes are reproduced in models and to further understand the carbon cycle in the real world.

Group dynamics challenges: Insights from Biosphere 2 experiments.

PubMed

Nelson, Mark; Gray, Kathelin; Allen, John P

2015-07-01

Successfully managing group dynamics of small, physically isolated groups is vital for long duration space exploration/habitation and for terrestrial CELSS (Controlled Environmental Life Support System) facilities with human participants. Biosphere 2 had important differences and shares some key commonalities with both Antarctic and space environments. There were a multitude of stress factors during the first two year closure experiment as well as mitigating factors. A helpful tool used at Biosphere 2 was the work of W.R. Bion who identified two competing modalities of behavior in small groups. Task-oriented groups are governed by conscious acceptance of goals, reality-thinking in relation to time and resources, and intelligent management of challenges. The opposing unconscious mode, the "basic-assumption" ("group animal") group, manifests through Dependency/Kill the Leader, Fight/Flight and Pairing. These unconscious dynamics undermine and can defeat the task group's goal. The biospherians experienced some dynamics seen in other isolated teams: factions developing reflecting personal chemistry and disagreements on overall mission procedures. These conflicts were exacerbated by external power struggles which enlisted support of those inside. Nevertheless, the crew evolved a coherent, creative life style to deal with some of the deprivations of isolation. The experience of the first two year closure of Biosphere 2 vividly illustrates both vicissitudes and management of group dynamics. The crew overrode inevitable frictions to creatively manage both operational and research demands and opportunities of the facility, thus staying 'on task' in Bion's group dynamics terminology. The understanding that Biosphere 2 was their life support system may also have helped the mission to succeed. Insights from the Biosphere 2 experience can help space and remote missions cope successfully with the inherent challenges of small, isolated crews. Copyright © 2015 The Committee on Space Research (COSPAR). Published by Elsevier Ltd. All rights reserved.

The Race for Space: Tracking Land-Cover Transformation in a Socio-ecological Landscape, South Africa

NASA Astrophysics Data System (ADS)

Coetzer, Kaera L.; Erasmus, Barend F. N.; Witkowski, Edward T. F.; Reyers, Belinda

2013-09-01

Biosphere Reserves attempt to align existing biodiversity conservation with sustainable resource use, specifically for improving socio-economic circumstances of resident communities. Typically, the Biosphere Reserve model is applied to an established landscape mosaic of existing land uses; these are often socio-ecological systems where strict environmental protection and community livelihoods are in conflict, and environmental degradation frequently accompanies "use". This raises challenges for successful implementation of the model, as the reality of the existing land-use mosaic undermines the theoretical aspirations of the Biosphere concept. This study focuses on the Kruger to Canyons Biosphere Reserve (K2C), South Africa; a socio-ecological landscape where formal conservation is juxtaposed against extensive impoverished rural communities. We focus on land-cover changes of the existing land-use mosaic (1993-2006), specifically selected land-cover classes identified as important for biodiversity conservation and local-level resource utilization. We discuss the implications of transformation for conservation, sustainable resource-use, and K2C's functioning as a "Biosphere Reserve". Spatially, changes radiated outward from the settlement expanse, with little regard for the theoretical land-use zonation of the Biosphere Reserve. Settlement growth tracked transport routes, transforming cohesive areas of communal-use rangelands. Given the interdependencies between the settlement population and local environmental resources, the Impacted Vegetation class expanded accordingly, fragmenting the Intact Vegetation class, and merging rangelands. This has serious implications for sustainability of communal harvesting areas, and further transformation of intact habitat. The distribution and magnitude of Intact Vegetation losses raise concerns around connectivity and edge effects, with long-term consequences for ecological integrity of remnant habitat, and K2C's existing network of protected areas.

On the Ability of Space- Based Passive and Active Remote Sensing Observations of CO2 to Detect Flux Perturbations to the Carbon Cycle

NASA Technical Reports Server (NTRS)

Crowell, Sean M. R.; Kawa, S. Randolph; Browell, Edward V.; Hammerling, Dorit M.; Moore, Berrien; Schaefer, Kevin; Doney, Scott C.

2018-01-01

Space-borne observations of CO2 are vital to gaining understanding of the carbon cycle in regions of the world that are difficult to measure directly, such as the tropical terrestrial biosphere, the high northern and southern latitudes, and in developing nations such as China. Measurements from passive instruments such as GOSAT (Greenhouse Gases Observing Satellite) and OCO-2 (Orbiting Carbon Observatory 2), however, are constrained by solar zenith angle limitations as well as sensitivity to the presence of clouds and aerosols. Active measurements such as those in development for the Active Sensing of CO2 Emissions over Nights, Days and Seasons (ASCENDS) mission show strong potential for making measurements in the high-latitude winter and in cloudy regions. In this work we examine the enhanced flux constraint provided by the improved coverage from an active measurement such as ASCENDS. The simulation studies presented here show that with sufficient precision, ASCENDS will detect permafrost thaw and fossil fuel emissions shifts at annual and seasonal time scales, even in the presence of transport errors, representativeness errors, and biogenic flux errors. While OCO-2 can detect some of these perturbations at the annual scale, the seasonal sampling provided by ASCENDS provides the stronger constraint. Plain Language Summary: Active and passive remote sensors show the potential to provide unprecedented information on the carbon cycle. With the all-season sampling, active remote sensors are more capable of constraining high-latitude emissions. The reduced sensitivity to cloud and aerosol also makes active sensors more capable of providing information in cloudy and polluted scenes with sufficient accuracy. These experiments account for errors that are fundamental to the top-down approach for constraining emissions, and even including these sources of error, we show that satellite remote sensors are critical for understanding the carbon cycle.

Patterns of new versus recycled primary production in the terrestrial biosphere

USDA-ARS?s Scientific Manuscript database

Nitrogen (N) and phosphorus (P) availability regulate plant productivity throughout the terrestrial biosphere, influencing the patterns and magnitude of net primary production (NPP) by land plants both now and into the future. These nutrients enter ecosystems via geologic and atmospheric pathways, a...

A biosphere assessment of high-level radioactive waste disposal in Sweden.

PubMed

Kautsky, Ulrik; Lindborg, Tobias; Valentin, Jack

2015-04-01

Licence applications to build a repository for the disposal of Swedish spent nuclear fuel have been lodged, underpinned by myriad reports and several broader reviews. This paper sketches out the technical and administrative aspects and highlights a recent review of the biosphere effects of a potential release from the repository. A comprehensive database and an understanding of major fluxes and pools of water and organic matter in the landscape let one envisage the future by looking at older parts of the site. Thus, today's biosphere is used as a natural analogue of possible future landscapes. It is concluded that the planned repository can meet the safety criteria and will have no detectable radiological impact on plants and animals. This paper also briefly describes biosphere work undertaken after the review. The multidisciplinary approach used is relevant in a much wider context and may prove beneficial across many environmental contexts. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Technical adequacy of bisulfite sequencing and pyrosequencing for detection of mitochondrial DNA methylation: Sources and avoidance of false-positive detection.

PubMed

Owa, Chie; Poulin, Matthew; Yan, Liying; Shioda, Toshi

2018-01-01

The existence of cytosine methylation in mammalian mitochondrial DNA (mtDNA) is a controversial subject. Because detection of DNA methylation depends on resistance of 5'-modified cytosines to bisulfite-catalyzed conversion to uracil, examined parameters that affect technical adequacy of mtDNA methylation analysis. Negative control amplicons (NCAs) devoid of cytosine methylation were amplified to cover the entire human or mouse mtDNA by long-range PCR. When the pyrosequencing template amplicons were gel-purified after bisulfite conversion, bisulfite pyrosequencing of NCAs did not detect significant levels of bisulfite-resistant cytosines (brCs) at ND1 (7 CpG sites) or CYTB (8 CpG sites) genes (CI95 = 0%-0.94%); without gel-purification, significant false-positive brCs were detected from NCAs (CI95 = 4.2%-6.8%). Bisulfite pyrosequencing of highly purified, linearized mtDNA isolated from human iPS cells or mouse liver detected significant brCs (~30%) in human ND1 gene when the sequencing primer was not selective in bisulfite-converted and unconverted templates. However, repeated experiments using a sequencing primer selective in bisulfite-converted templates almost completely (< 0.8%) suppressed brC detection, supporting the false-positive nature of brCs detected using the non-selective primer. Bisulfite-seq deep sequencing of linearized, gel-purified human mtDNA detected 9.4%-14.8% brCs for 9 CpG sites in ND1 gene. However, because all these brCs were associated with adjacent non-CpG brCs showing the same degrees of bisulfite resistance, DNA methylation in this mtDNA-encoded gene was not confirmed. Without linearization, data generated by bisulfite pyrosequencing or deep sequencing of purified mtDNA templates did not pass the quality control criteria. Shotgun bisulfite sequencing of human mtDNA detected extremely low levels of CpG methylation (<0.65%) over non-CpG methylation (<0.55%). Taken together, our study demonstrates that adequacy of mtDNA methylation analysis using methods dependent on bisulfite conversion needs to be established for each experiment, taking effects of incomplete bisulfite conversion and template impurity or topology into consideration.

Towards understanding the variability in biospheric CO2 fluxes: using FTIR spectrometry and a chemical transport model to investigate the sources and sinks of carbonyl sulfide and its link to CO2

NASA Astrophysics Data System (ADS)

Wang, Y.; Deutscher, N. M.; Palm, M.; Warneke, T.; Notholt, J.; Baker, I.; Berry, J.; Suntharalingam, P.; Jones, N.; Mahieu, E.; Lejeune, B.; Campbell, J. E.; Wolf, A.; Kremser, S.

2015-09-01

Understanding carbon dioxide (CO2) biospheric processes is of great importance because the terrestrial exchange drives the seasonal and inter-annual variability of CO2 in the atmosphere. Atmospheric inversions based on CO2 concentration measurements alone can only determine net biosphere fluxes, but not differentiate between photosynthesis (uptake) and respiration (production). Carbonyl sulfide (OCS) could provide an important additional constraint: it is also taken up by plants during photosynthesis but not emitted during respiration, and therefore is a potential mean to differentiate between these processes. Solar absorption Fourier Transform InfraRed (FTIR) spectrometry allows for the retrievals of the atmospheric concentrations of both CO2 and OCS from measured solar absorption spectra. Here, we investigate co-located and quasi-simultaneous FTIR measurements of OCS and CO2 performed at three selected sites located in the Northern Hemisphere. These measurements are compared to simulations of OCS and CO2 using a chemical transport model (GEOS-Chem). The OCS simulations are driven by different land biospheric fluxes to reproduce the seasonality of the measurements. Increasing the plant uptake of Kettle et al. (2002a) by a factor of three resulted in the best comparison with FTIR measurements. However, there are still discrepancies in the latitudinal distribution when comparing with HIPPO (HIAPER Pole-to-Pole Observations) data spanning both hemispheres. The coupled biospheric fluxes of OCS and CO2 from the simple biosphere model (SiB) are used in the study and compared to measurements. The CO2 simulation with SiB fluxes agrees with the measurements well, while the OCS simulation reproduced a weaker drawdown than FTIR measurements at selected sites, and a smaller latitudinal gradient in the Northern Hemisphere during growing season. An offset in the timing of the seasonal cycle minimum between SiB simulation and measurements is also seen. Using OCS as a photosynthesis proxy can help to understand how the biospheric processes are reproduced in models and to further understand the carbon cycle in the real world.

Quantifying the influence of the terrestrial biosphere on glacial-interglacial climate dynamics

NASA Astrophysics Data System (ADS)

Davies-Barnard, Taraka; Ridgwell, Andy; Singarayer, Joy; Valdes, Paul

2017-10-01

The terrestrial biosphere is thought to be a key component in the climatic variability seen in the palaeo-record. It has a direct impact on surface temperature through changes in surface albedo and evapotranspiration (so-called biogeophysical effects) and, in addition, has an important indirect effect through changes in vegetation and soil carbon storage (biogeochemical effects) and hence modulates the concentrations of greenhouse gases in the atmosphere. The biogeochemical and biogeophysical effects generally have opposite signs, meaning that the terrestrial biosphere could potentially have played only a very minor role in the dynamics of the glacial-interglacial cycles of the late Quaternary. Here we use a fully coupled dynamic atmosphere-ocean-vegetation general circulation model (GCM) to generate a set of 62 equilibrium simulations spanning the last 120 kyr. The analysis of these simulations elucidates the relative importance of the biogeophysical versus biogeochemical terrestrial biosphere interactions with climate. We find that the biogeophysical effects of vegetation account for up to an additional -0.91 °C global mean cooling, with regional cooling as large as -5 °C, but with considerable variability across the glacial-interglacial cycle. By comparison, while opposite in sign, our model estimates of the biogeochemical impacts are substantially smaller in magnitude. Offline simulations show a maximum of +0.33 °C warming due to an increase of 25 ppm above our (pre-industrial) baseline atmospheric CO2 mixing ratio. In contrast to shorter (century) timescale projections of future terrestrial biosphere response where direct and indirect responses may at times cancel out, we find that the biogeophysical effects consistently and strongly dominate the biogeochemical effect over the inter-glacial cycle. On average across the period, the terrestrial biosphere has a -0.26 °C effect on temperature, with -0.58 °C at the Last Glacial Maximum. Depending on assumptions made about the destination of terrestrial carbon under ice sheets and where sea level has changed, the average terrestrial biosphere contribution over the last 120 kyr could be as much as -50 °C and -0.83 °C at the Last Glacial Maximum.

Exploring frontiers of the deep biosphere through scientific ocean drilling

NASA Astrophysics Data System (ADS)

Inagaki, F.; D'Hondt, S.; Hinrichs, K. U.

2015-12-01

Since the first deep biosphere-dedicated Ocean Drilling Program (ODP) Leg 201 using the US drill ship JOIDES Resolution in 2002, scientific ocean drilling has offered unique opportunities to expand our knowledge of the nature and extent of the deep biosphere. The latest estimate of the global subseafloor microbial biomass is ~1029cells, accounting for 4 Gt of carbon and ~1% of the Earth's total living biomass. The subseafloor microbial communities are evolutionarily diverse and their metabolic rates are extraordinarily slow. Nevertheless, accumulating activity most likely plays a significant role in elemental cycles over geological time. In 2010, during Integrated Ocean Drilling Program (IODP) Expedition 329, the JOIDES Resolutionexplored the deep biosphere in the open-ocean South Pacific Gyre—the largest oligotrophic province on our planet. During Expedition 329, relatively high concentrations of dissolved oxygen and significantly low biomass of microbial populations were observed in the entire sediment column, indicating that (i) there is no limit to life in open-ocean sediment and (ii) a significant amount of oxygen reaches through the sediment to the upper oceanic crust. This "deep aerobic biosphere" inhabits the sediment throughout up to ~37 percent of the world's oceans. The remaining ~63 percent of the oceans is comprised of higher productivity areas that contain the "deep anaerobic biosphere". In 2012, during IODP Expedition 337, the Japanese drill ship Chikyu explored coal-bearing sediments down to 2,466 meters below the seafloor off the Shimokita Peninsula, Japan. Geochemical and microbiological analyses consistently showed the occurrence of methane-producing communities associated with the coal beds. Cell concentrations in deep sediments were notably lower than those expected from the global regression line, implying that the bottom of the deep biosphere is approached in these beds. Taxonomic composition of the deep coal-bearing communities profoundly differs from those in shallower marine sediments and instead resembles organotrophic communities in forest soils. These findings suggest that the terrigenous microbial ecosystem has been partly retained from the original depositional setting over 20 million years and contributed to deep carbon cycling ever since.

Intracellular diversity of the V4 and V9 regions of the 18S rRNA in marine protists (radiolarians) assessed by high-throughput sequencing.

PubMed

Decelle, Johan; Romac, Sarah; Sasaki, Eriko; Not, Fabrice; Mahé, Frédéric

2014-01-01

Metabarcoding is a powerful tool for exploring microbial diversity in the environment, but its accurate interpretation is impeded by diverse technical (e.g. PCR and sequencing errors) and biological biases (e.g. intra-individual polymorphism) that remain poorly understood. To help interpret environmental metabarcoding datasets, we investigated the intracellular diversity of the V4 and V9 regions of the 18S rRNA gene from Acantharia and Nassellaria (radiolarians) using 454 pyrosequencing. Individual cells of radiolarians were isolated, and PCRs were performed with generalist primers to amplify the V4 and V9 regions. Different denoising procedures were employed to filter the pyrosequenced raw amplicons (Acacia, AmpliconNoise, Linkage method). For each of the six isolated cells, an average of 541 V4 and 562 V9 amplicons assigned to radiolarians were obtained, from which one numerically dominant sequence and several minor variants were found. At the 97% identity, a diversity metrics commonly used in environmental surveys, up to 5 distinct OTUs were detected in a single cell. However, most amplicons grouped within a single OTU whereas other OTUs contained very few amplicons. Different analytical methods provided evidence that most minor variants forming different OTUs correspond to PCR and sequencing artifacts. Duplicate PCR and sequencing from the same DNA extract of a single cell had only 9 to 16% of unique amplicons in common, and alignment visualization of V4 and V9 amplicons showed that most minor variants contained substitutions in highly-conserved regions. We conclude that intracellular variability of the 18S rRNA in radiolarians is very limited despite its multi-copy nature and the existence of multiple nuclei in these protists. Our study recommends some technical guidelines to conservatively discard artificial amplicons from metabarcoding datasets, and thus properly assess the diversity and richness of protists in the environment.

Bacterial Population in Intestines of the Black Tiger Shrimp (Penaeus monodon) under Different Growth Stages

PubMed Central

Rungrassamee, Wanilada; Klanchui, Amornpan; Chaiyapechara, Sage; Maibunkaew, Sawarot; Tangphatsornruang, Sithichoke; Jiravanichpaisal, Pikul; Karoonuthaisiri, Nitsara

2013-01-01

Intestinal bacterial communities in aquaculture have been drawn to attention due to potential benefit to their hosts. To identify core intestinal bacteria in the black tiger shrimp (Penaeus monodon), bacterial populations of disease-free shrimp were characterized from intestines of four developmental stages (15-day-old post larvae (PL15), 1- (J1), 2- (J2), and 3-month-old (J3) juveniles) using pyrosequencing, real-time PCR and denaturing gradient gel electrophoresis (DGGE) approaches. A total of 25,121 pyrosequencing reads (reading length = 442±24 bases) were obtained, which were categorized by barcode for PL15 (7,045 sequences), J1 (3,055 sequences), J2 (13,130 sequences) and J3 (1,890 sequences). Bacteria in the phyla Bacteroides, Firmicutes and Proteobacteria were found in intestines at all four growth stages. There were 88, 14, 27, and 20 bacterial genera associated with the intestinal tract of PL15, J1, J2 and J3, respectively. Pyrosequencing analysis revealed that Proteobacteria (class Gammaproteobacteria) was a dominant bacteria group with a relative abundance of 89% for PL15 and 99% for J1, J2 and J3. Real-time PCR assay also confirmed that Gammaproteobacteria had the highest relative abundance in intestines from all growth stages. Intestinal bacterial communities from the three juvenile stages were more similar to each other than that of the PL shrimp based on PCA analyses of pyrosequencing results and their DGGE profiles. This study provides descriptive bacterial communities associated to the black tiger shrimp intestines during these growth development stages in rearing facilities. PMID:23577162

Bacterial population in intestines of the black tiger shrimp (Penaeus monodon) under different growth stages.

PubMed

Rungrassamee, Wanilada; Klanchui, Amornpan; Chaiyapechara, Sage; Maibunkaew, Sawarot; Tangphatsornruang, Sithichoke; Jiravanichpaisal, Pikul; Karoonuthaisiri, Nitsara

2013-01-01

Intestinal bacterial communities in aquaculture have been drawn to attention due to potential benefit to their hosts. To identify core intestinal bacteria in the black tiger shrimp (Penaeus monodon), bacterial populations of disease-free shrimp were characterized from intestines of four developmental stages (15-day-old post larvae (PL15), 1- (J1), 2- (J2), and 3-month-old (J3) juveniles) using pyrosequencing, real-time PCR and denaturing gradient gel electrophoresis (DGGE) approaches. A total of 25,121 pyrosequencing reads (reading length = 442±24 bases) were obtained, which were categorized by barcode for PL15 (7,045 sequences), J1 (3,055 sequences), J2 (13,130 sequences) and J3 (1,890 sequences). Bacteria in the phyla Bacteroides, Firmicutes and Proteobacteria were found in intestines at all four growth stages. There were 88, 14, 27, and 20 bacterial genera associated with the intestinal tract of PL15, J1, J2 and J3, respectively. Pyrosequencing analysis revealed that Proteobacteria (class Gammaproteobacteria) was a dominant bacteria group with a relative abundance of 89% for PL15 and 99% for J1, J2 and J3. Real-time PCR assay also confirmed that Gammaproteobacteria had the highest relative abundance in intestines from all growth stages. Intestinal bacterial communities from the three juvenile stages were more similar to each other than that of the PL shrimp based on PCA analyses of pyrosequencing results and their DGGE profiles. This study provides descriptive bacterial communities associated to the black tiger shrimp intestines during these growth development stages in rearing facilities.

Lyssavirus Detection and Typing Using Pyrosequencing▿#‖

PubMed Central

De Benedictis, Paola; De Battisti, Cristian; Dacheux, Laurent; Marciano, Sabrina; Ormelli, Silvia; Salomoni, Angela; Caenazzo, Silvia Tiozzo; Lepelletier, Anthony; Bourhy, Hervé; Capua, Ilaria; Cattoli, Giovanni

2011-01-01

Rabies is a fatal zoonosis caused by a nonsegmented negative-strand RNA virus, namely, rabies virus (RABV). Apart from RABV, at least 10 additional species are known as rabies-related lyssaviruses (RRVs), and some of them are responsible for occasional spillovers into humans. More lyssaviruses have also been detected recently in different bat ecosystems, thanks to the application of molecular diagnostic methods. Due to the variety of the members of the genus Lyssavirus, there is the necessity to develop a reliable molecular assay for rabies diagnosis able to detect and differentiate among the existing rabies and rabies-related viruses. In the present study, a pyrosequencing protocol targeting the 3′ terminus of the nucleoprotein (N) gene was applied for the rapid characterization of lyssaviruses. Correct identification of species was achieved for each sample tested. Results from the pyrosequencing assay were also confirmed by those obtained using the Sanger sequencing method. A pan-lyssavirus one-step reverse transcription (RT)-PCR was developed within the framework of the pyrosequencing procedure. The sensitivity (Se) of the one-step RT-PCR assay was determined by using in vitro-transcribed RNA and serial dilutions of titrated viruses. The assay demonstrated high analytical and relative specificity (Sp) (98.94%) and sensitivity (99.71%). To date, this is the first case in which pyrosequencing has been applied for lyssavirus identification using a cheaper diagnostic approach than the one for all the other protocols for rapid typing that we are acquainted with. Results from this study indicate that this procedure is suitable for lyssavirus detection in samples of both human and animal origin. PMID:21389152

Application of Pyrosequencing® in Food Biodefense.

PubMed

Amoako, Kingsley Kwaku

2015-01-01

The perpetration of a bioterrorism attack poses a significant risk for public health with potential socioeconomic consequences. It is imperative that we possess reliable assays for the rapid and accurate identification of biothreat agents to make rapid risk-informed decisions on emergency response. The development of advanced methodologies for the detection of biothreat agents has been evolving rapidly since the release of the anthrax spores in the mail in 2001, and recent advances in detection and identification techniques could prove to be an essential component in the defense against biological attacks. Sequence-based approaches such as Pyrosequencing(®), which has the capability to determine short DNA stretches in real time using biotinylated PCR amplicons, have potential biodefense applications. Using markers from the virulence plasmids and chromosomal regions, my laboratory has demonstrated the power of this technology in the rapid, specific, and sensitive detection of B. anthracis spores and Yersinia pestis in food. These are the first applications for the detection of the two organisms in food. Furthermore, my lab has developed a rapid assay to characterize the antimicrobial resistance (AMR) gene profiles for Y. pestis using Pyrosequencing. Pyrosequencing is completed in about 60 min (following PCR amplification) and yields accurate and reliable results with an added layer of confidence, thus enabling rapid risk-informed decisions to be made. A typical run yields 40-84 bp reads with 94-100 % identity to the expected sequence. It also provides a rapid method for determining the AMR profile as compared to the conventional plate method which takes several days. The method described is proposed as a novel detection system for potential application in food biodefense.

Field Monitoring of Avian Influenza Viruses: Whole-Genome Sequencing and Tracking of Neuraminidase Evolution Using 454 Pyrosequencing

PubMed Central

Croville, Guillaume; Soubies, Sébastien Mathieu; Barbieri, Johanna; Klopp, Christophe; Mariette, Jérôme; Bouchez, Olivier; Camus-Bouclainville, Christelle

2012-01-01

Adaptation of avian influenza viruses (AIVs) from waterfowl to domestic poultry with a deletion in the neuraminidase (NA) stalk has already been reported. The way the virus undergoes this evolution, however, is thus far unclear. We address this question using pyrosequencing of duck and turkey low-pathogenicity AIVs. Ducks and turkeys were sampled at the very beginning of an H6N1 outbreak, and turkeys were swabbed again 8 days later. NA stalk deletions were evidenced in turkeys by Sanger sequencing. To further investigate viral evolution, 454 pyrosequencing was performed: for each set of samples, up to 41,500 reads of ca. 400 bp were generated and aligned. Genetic polymorphisms between duck and turkey viruses were tracked on the whole genome. NA deletion was detected in less than 2% of reads in duck feces but in 100% of reads in turkey tracheal specimens collected at the same time. Further variations in length were observed in NA from turkeys 8 days later. Similarly, minority mutants emerged on the hemagglutinin (HA) gene, with substitutions mostly in the receptor binding site on the globular head. These critical changes suggest a strong evolutionary pressure in turkeys. The increasing performances of next-generation sequencing technologies should enable us to monitor the genomic diversity of avian influenza viruses and early emergence of potentially pathogenic variants within bird flocks. The present study, based on 454 pyrosequencing, suggests that NA deletion, an example of AIV adaptation from waterfowl to domestic poultry, occurs by selection rather than de novo emergence of viral mutants. PMID:22718944

Metatranscriptomics and Pyrosequencing Facilitate Discovery of Potential Viral Natural Enemies of the Invasive Caribbean Crazy Ant, Nylanderia pubens

PubMed Central

Valles, Steven M.; Oi, David H.; Yu, Fahong; Tan, Xin-Xing; Buss, Eileen A.

2012-01-01

Background Nylanderia pubens (Forel) is an invasive ant species that in recent years has developed into a serious nuisance problem in the Caribbean and United States. A rapidly expanding range, explosive localized population growth, and control difficulties have elevated this ant to pest status. Professional entomologists and the pest control industry in the United States are urgently trying to understand its biology and develop effective control methods. Currently, no known biological-based control agents are available for use in controlling N. pubens. Methodology and Principal Findings Metagenomics and pyrosequencing techniques were employed to examine the transcriptome of field-collected N. pubens colonies in an effort to identify virus infections with potential to serve as control agents against this pest ant. Pyrosequencing (454-platform) of a non-normalized N. pubens expression library generated 1,306,177 raw sequence reads comprising 450 Mbp. Assembly resulted in generation of 59,017 non-redundant sequences, including 27,348 contigs and 31,669 singlets. BLAST analysis of these non-redundant sequences identified 51 of potential viral origin. Additional analyses winnowed this list of potential viruses to three that appear to replicate in N. pubens. Conclusions Pyrosequencing the transcriptome of field-collected samples of N. pubens has identified at least three sequences that are likely of viral origin and, in which, N. pubens serves as host. In addition, the N. pubens transcriptome provides a genetic resource for the scientific community which is especially important at this early stage of developing a knowledgebase for this new pest. PMID:22384082

CYP3A4 and CYP3A5 genotyping by Pyrosequencing

PubMed Central

Garsa, Adam A; McLeod, Howard L; Marsh, Sharon

2005-01-01

Background Human cytochrome P450 3A enzymes, particularly CYP3A4 and CYP3A5, play an important role in drug metabolism. CYP3A expression exhibits substantial interindividual variation, much of which may result from genetic variation. This study describes Pyrosequencing assays for key SNPs in CYP3A4 (CYP3A4*1B, CYP3A4*2, and CYP3A4*3) and CYP3A5 (CYP3A5*3C and CYP3A5*6). Methods Genotyping of 95 healthy European and 95 healthy African volunteers was performed using Pyrosequencing. Linkage disequilibrium, haplotype inference, Hardy-Weinberg equilibrium, and tag SNPs were also determined for these samples. Results CYP3A4*1B allele frequencies were 4% in Europeans and 82% in Africans. The CYP3A4*2 allele was found in neither population sample. CYP3A4*3 had an allele frequency of 2% in Europeans and 0% in Africans. The frequency of CYP3A5*3C was 94% in Europeans and 12% in Africans. No CYP3A5*6 variants were found in the European samples, but this allele had a frequency of 16% in the African samples. Allele frequencies and haplotypes show interethnic variation, highlighting the need to analyze clinically relevant SNPs and haplotypes in a variety of ethnic groups. Conclusion Pyrosequencing is a versatile technique that could improve the efficiency of SNP analysis for pharmacogenomic research with the ultimate goal of pre-screening patients for individual therapy selection. PMID:15882469

Access 1996: A directory of permanent plots which monitor flora, fauna, climate, hydrology, soil, geology, and the effects of anthropogenic changes at 132 biosphere reserves in 27 countries

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

1996-07-01

This directory summarizes information about environmental data collected in permanent monitoring and research plots in 132 biosphere reserves in Canada, the United States, and 25 European countries. The text of the directory is organized alphabetically by country and, within each country, alphabetically according to the name of the biosphere reserve. Tabular summaries of information on permanent plots are provided. The summaries are organized topically . A general summary of basic information on permanent plots is followed by more detailed information on permanent plots dedicated primarily to monitroing and research on particular topics.

10 CFR 63.305 - Required characteristics of the reference biosphere.

Code of Federal Regulations, 2013 CFR

2013-01-01

... the region surrounding the Yucca Mountain site. (b) DOE should not project changes in society, the biosphere (other than climate), human biology, or increases or decreases of human knowledge or technology... vary factors related to the geology, hydrology, and climate based upon cautious, but reasonable...

10 CFR 63.305 - Required characteristics of the reference biosphere.

Code of Federal Regulations, 2014 CFR

2014-01-01

... the region surrounding the Yucca Mountain site. (b) DOE should not project changes in society, the biosphere (other than climate), human biology, or increases or decreases of human knowledge or technology... vary factors related to the geology, hydrology, and climate based upon cautious, but reasonable...

10 CFR 63.305 - Required characteristics of the reference biosphere.

Code of Federal Regulations, 2012 CFR

2012-01-01

... the region surrounding the Yucca Mountain site. (b) DOE should not project changes in society, the biosphere (other than climate), human biology, or increases or decreases of human knowledge or technology... vary factors related to the geology, hydrology, and climate based upon cautious, but reasonable...

10 CFR 63.305 - Required characteristics of the reference biosphere.

Code of Federal Regulations, 2010 CFR

2010-01-01

... the region surrounding the Yucca Mountain site. (b) DOE should not project changes in society, the biosphere (other than climate), human biology, or increases or decreases of human knowledge or technology... vary factors related to the geology, hydrology, and climate based upon cautious, but reasonable...

Reaching for Excellence.

ERIC Educational Resources Information Center

Wright, Emmett L.; Perna, Jack A.

1992-01-01

Presents the four program goals for biology set forth in the National Science Teacher Association's "A Focus on Excellence: Biology Revisited" to (1) address biosphere, human society, and individual needs; (2) encourage students to experience, understand, and appreciate of natural systems; (3) apply the basic concept of the biosphere; and (4)…

The World Campaign for the Biosphere.

ERIC Educational Resources Information Center

Barman, Charles R.

1984-01-01

Lists and discusses goals of The World Campaign for the Biosphere and strategies designed to achieve these goals. Also lists eight suggestions for science teachers to help incorporate the goals into school curricula and programs. These include organizing assemblies which present information about environmental problems and presenting environmental…

Viral infections as controlling factors for the deep biosphere? (Invited)

NASA Astrophysics Data System (ADS)

Engelen, B.; Engelhardt, T.; Sahlberg, M.; Cypionka, H.

2009-12-01

The marine deep biosphere represents the largest biotope on Earth. Throughout the last years, we have obtained interesting insights into its microbial community composition. However, one component that was completely overlooked so far is the viral inventory of deep-subsurface sediments. While viral infections were identified to have a major impact on the benthic microflora of deep-sea surface sediments (Danavaro et al. 2008), no studies were performed on deep-biosphere samples, so far. As grazers probably play only a minor role in anoxic and highly compressed deep sediments, viruses might be the main “predators” for indigenous microorganisms. Furthermore, the release of cell components, called “the viral shunt”, could have a major impact on the deep biosphere in providing labile organic compounds to non-infected microorganisms in these generally nutrient depleted sediments. However, direct counting of viruses in sediments is highly challenging due to the small size of viruses and the high background of small particles. Even molecular surveys using “universal” PCR primers that target phage-specific genes fail due to the vast phage diversity. One solution for this problem is the lysogenic viral life cycle as many bacteriophages integrate their DNA into the host genome. It is estimated that up to 70% of cultivated bacteria contain prophages within their genome. Therefore, culture collections (Batzke et al. 2007) represent an archive of the viral composition within the respective habitat. These prophages can be induced to become free phage particles in stimulation experiments in which the host cells are set under certain stress situations such as a treatment with UV exposure or DNA-damaging antibiotics. The study of the viral component within the deep biosphere offers to answer the following questions: To which extent are deep-biosphere populations controlled by viral infections? What is the inter- and intra-specific diversity and the host-specific viral biogeography? Can viral infections tell us something about the physiological state of indigenous microorganisms? Finally, we will obtain estimates for the viral shunt as an important factor for sustaining the deep biosphere. References: Batzke A, Engelen B, Sass H, Cypionka H (2007) Phylogenetic and physiological diversity of cultured deep-biosphere bacteria from Equatorial Pacific Ocean and Peru Margin sediments. Geomicrobiology J 24:261-273 Danovaro R, Dell'Anno A, Corinaldesi C, Magagnini M, Noble R, Tamburini C, Weinbauer M (2008) Major viral impact on the functioning of benthic deep-sea ecosystems. Nature 454: 1084-U1027.

454-Pyrosequencing Analysis of Bacterial Communities from Autotrophic Nitrogen Removal Bioreactors Utilizing Universal Primers: Effect of Annealing Temperature

PubMed Central

Rodriguez-Sanchez, Alejandro; Rodelas, Belén; Abbas, Ben A.; Martinez-Toledo, Maria Victoria; van Loosdrecht, Mark C. M.; Osorio, F.; Gonzalez-Lopez, Jesus

2015-01-01

Identification of anaerobic ammonium oxidizing (anammox) bacteria by molecular tools aimed at the evaluation of bacterial diversity in autotrophic nitrogen removal systems is limited by the difficulty to design universal primers for the Bacteria domain able to amplify the anammox 16S rRNA genes. A metagenomic analysis (pyrosequencing) of total bacterial diversity including anammox population in five autotrophic nitrogen removal technologies, two bench-scale models (MBR and Low Temperature CANON) and three full-scale bioreactors (anammox, CANON, and DEMON), was successfully carried out by optimization of primer selection and PCR conditions (annealing temperature). The universal primer 530F was identified as the best candidate for total bacteria and anammox bacteria diversity coverage. Salt-adjusted optimum annealing temperature of primer 530F was calculated (47°C) and hence a range of annealing temperatures of 44–49°C was tested. Pyrosequencing data showed that annealing temperature of 45°C yielded the best results in terms of species richness and diversity for all bioreactors analyzed. PMID:26421306

454-Pyrosequencing Analysis of Bacterial Communities from Autotrophic Nitrogen Removal Bioreactors Utilizing Universal Primers: Effect of Annealing Temperature.

PubMed

Gonzalez-Martinez, Alejandro; Rodriguez-Sanchez, Alejandro; Rodelas, Belén; Abbas, Ben A; Martinez-Toledo, Maria Victoria; van Loosdrecht, Mark C M; Osorio, F; Gonzalez-Lopez, Jesus

2015-01-01

Identification of anaerobic ammonium oxidizing (anammox) bacteria by molecular tools aimed at the evaluation of bacterial diversity in autotrophic nitrogen removal systems is limited by the difficulty to design universal primers for the Bacteria domain able to amplify the anammox 16S rRNA genes. A metagenomic analysis (pyrosequencing) of total bacterial diversity including anammox population in five autotrophic nitrogen removal technologies, two bench-scale models (MBR and Low Temperature CANON) and three full-scale bioreactors (anammox, CANON, and DEMON), was successfully carried out by optimization of primer selection and PCR conditions (annealing temperature). The universal primer 530F was identified as the best candidate for total bacteria and anammox bacteria diversity coverage. Salt-adjusted optimum annealing temperature of primer 530F was calculated (47°C) and hence a range of annealing temperatures of 44-49°C was tested. Pyrosequencing data showed that annealing temperature of 45°C yielded the best results in terms of species richness and diversity for all bioreactors analyzed.

Detection of Drug-Resistant Mycobacterium tuberculosis.

PubMed

Engström, Anna; Juréen, Pontus

2015-01-01

Tuberculosis (TB) remains a global health problem. The increasing prevalence of drug-resistant Mycobacterium tuberculosis, the causative agent of TB, demands new measures to combat the situation. Rapid and accurate diagnosis of the pathogen and its drug susceptibility pattern is essential for timely initiation of optimal treatment, and, ultimately, control of the disease. We have developed a molecular method for detection of first- and second-line drug resistance in M. tuberculosis by Pyrosequencing(®). The method consists of seven Pyrosequencing assays for the detection of mutations in the genes or promoter regions, which are most commonly responsible for resistance to the drugs rifampicin, isoniazid, ethambutol, amikacin, kanamycin, capreomycin, and fluoroquinolones. The method was validated on clinical isolates and it was shown that the sensitivity and specificity of the method were comparable to those of Sanger sequencing. In the protocol in this chapter we describe the steps necessary for setting up and performing Pyrosequencing for M. tuberculosis. The first part of the protocol describes the assay development and the second part of the protocol describes utilization of the method.

Self-organization of the earth's biosphere-geochemical or geophysiological?

NASA Technical Reports Server (NTRS)

Schwartzman, David W.; Shore, Steven N.; Volk, Tyler; Mcmenamin, Mark

1994-01-01

We explore the implications of indicating the biosphere's self-organization by the trend over time of the net entropic flow from the Earth's surface, the actual physical boundary of virtually all biotic mass. This flow, derived from the radiative surface entropy budget, is approximately inversely related to the surface temperature when the solar incident flux remains constant. In the geophysiological ('gaian') interpretation, biospheric self-organization has increased with the progressive colonization of the continents and evolutionary developments in the land biota, as a result of surface cooling arising from biotic enhancement of weathering. The key site for this self-organization is at the interface between land and atmosphere, the soil, where carbon is sequestered by its reaction (as carbonic and organic acids) with calcium magnesium silicates. Along with disequilibrium (steady-state) levels of carbon dioxide in the atmosphere, the occurrence of differentiated soil is the critical material evidence for biospheric self-organization, whether it be geophysiological or geochemical (ie., purely result of inorganic reactions). The computed equilibrium levels of carbon dioxide and corresponding equilibrium temperatures in the past are dramatically different from the steady-state levels. With future solar luminosity increase, the biospheric capacity for climatic regulation will decrease, leading to the ending of self-organization some two billion years from now. The Earth's surface will then approach chemical equilibrium with respect to the carbonate-silicate cycle.

Inhabitants' environmental perceptions in the city of Rome within the framework for urban biosphere reserves of the UNESCO programme on man and biosphere.

PubMed

Bonnes, Mirilia; Carrus, Giuseppe; Bonaiuto, Marino; Fornara, Ferdinando; Passafaro, Paola

2004-06-01

The article presents the main tenets of the UNESCO Programme on Man and Biosphere (MAB), launched by the United Nations at the beginning of the 1970s. The program aimed at supporting applied research and scientific knowledge for managing natural resources in a rational and sustainable way. The implication of the full ecological perspective, typical of the MAB, for promoting multidisciplinary and integrated approaches in the study of environmental issues is briefly outlined. In particular, we point out the role of the MAB, through the biosphere reserve concept, in supporting the collaboration between natural-biological and social-behavioral sciences when dealing with biodiversity conservation problems and with urban ecosystems. Then, the specific UNESCO-MAB Project on the city of Rome, launched at the end of the 1980s, is briefly presented, together with the recent project of the Department of the Environment of the Rome Municipality to propose Rome's urban and periurban green areas as a new UNESCO-MAB Biosphere Reserve. The results of the main research activities conducted therein are summarized. In particular, the specific research lines of the environmental psychology research group, involved in the MAB-Rome Project for approximately two decades, are presented. These research lines dealt with various aspects of residents' environmental perceptions and behaviors in the city of Rome. The practical implications of these results are also briefly discussed.

Impact of the 2008 Wenchuan earthquake on river organic carbon provenance: Insight from biomarkers

NASA Astrophysics Data System (ADS)

Wang, Jin; Feng, Xiaojuan; Hilton, Robert; Jin, Zhangdong; Ma, Tian; Zhang, Fei; Li, Gen; Densmore, Alexander; West, A. Joshua

2017-04-01

Large earthquakes can trigger widespread landslides in active mountain belts, which can mobilize biospheric organic carbon (OC) from the soil and vegetation. Rivers can erode and export biospheric particulate organic carbon (POC), which is an export of ecosystem productivity and may result in a CO2 sink if buried in sedimentary deposits. Our previous work showed that the 2008 Mw 7.9 Wenchuan earthquake increased the discharge of biospheric OC by rivers, due to the increased supply by earthquake triggered landslides (Wang et al., 2016). However, while the OC derived from sedimentary rocks could be accounted for, the source of biospheric OC in rivers before and after the earthquake remains poorly constrained. Here we use suspended sediment samples collected from the Zagunao River before and after the Wenchuan earthquake and measured the specific compounds of OC, including fatty acids, lignin phenols and glycerol dialkyl glycerol tetraether (GDGT) lipids. In combination with the analysis of bulk elemental concentration (C and N) and carbon isotopic ratio, the new data shows differential export patterns for OC components derived from varied terrestrial sources. A high frequency sampling enabled us to explore how the biospheric OC source changes following the earthquake, helping to better understand the link between active tectonics and the carbon cycle. Our results are also important in revealing how sedimentary biomarker records may record past earthquakes.

The Anthropocene: a conspicuous stratigraphical signal of anthropogenic changes in production and consumption across the biosphere

NASA Astrophysics Data System (ADS)

Williams, Mark; Zalasiewicz, Jan; Waters, Colin N.; Edgeworth, Matt; Bennett, Carys; Barnosky, Anthony D.; Ellis, Erle C.; Ellis, Michael A.; Cearreta, Alejandro; Haff, Peter K.; Ivar do Sul, Juliana A.; Leinfelder, Reinhold; McNeill, John R.; Odada, Eric; Oreskes, Naomi; Revkin, Andrew; Richter, Daniel deB; Steffen, Will; Summerhayes, Colin; Syvitski, James P.; Vidas, Davor; Wagreich, Michael; Wing, Scott L.; Wolfe, Alexander P.; Zhisheng, An

2016-03-01

Biospheric relationships between production and consumption of biomass have been resilient to changes in the Earth system over billions of years. This relationship has increased in its complexity, from localized ecosystems predicated on anaerobic microbial production and consumption to a global biosphere founded on primary production from oxygenic photoautotrophs, through the evolution of Eukarya, metazoans, and the complexly networked ecosystems of microbes, animals, fungi, and plants that characterize the Phanerozoic Eon (the last ˜541 million years of Earth history). At present, one species, Homo sapiens, is refashioning this relationship between consumption and production in the biosphere with unknown consequences. This has left a distinctive stratigraphy of the production and consumption of biomass, of natural resources, and of produced goods. This can be traced through stone tool technologies and geochemical signals, later unfolding into a diachronous signal of technofossils and human bioturbation across the planet, leading to stratigraphically almost isochronous signals developing by the mid-20th century. These latter signals may provide an invaluable resource for informing and constraining a formal Anthropocene chronostratigraphy, but are perhaps yet more important as tracers of a biosphere state that is characterized by a geologically unprecedented pattern of global energy flow that is now pervasively influenced and mediated by humans, and which is necessary for maintaining the complexity of modern human societies.

Comparative Viral Metagenomics of Environmental Samples from Korea

PubMed Central

Kim, Min-Soo; Whon, Tae Woong

2013-01-01

The introduction of metagenomics into the field of virology has facilitated the exploration of viral communities in various natural habitats. Understanding the viral ecology of a variety of sample types throughout the biosphere is important per se, but it also has potential applications in clinical and diagnostic virology. However, the procedures used by viral metagenomics may produce technical errors, such as amplification bias, while public viral databases are very limited, which may hamper the determination of the viral diversity in samples. This review considers the current state of viral metagenomics, based on examples from Korean viral metagenomic studies-i.e., rice paddy soil, fermented foods, human gut, seawater, and the near-surface atmosphere. Viral metagenomics has become widespread due to various methodological developments, and much attention has been focused on studies that consider the intrinsic role of viruses that interact with their hosts. PMID:24124407

Mars Exobiology: The Principles Behind The Plan For Exploration

NASA Technical Reports Server (NTRS)

DesMarais, D. J.; DeVincenzi, Donald L.; Carr, M. H.; Clark, B. C.; Farmer, J. D.; Hayes, J. M.; Holland, H.; Kerridge, J. F.; Klein, H. P.; McDonald, G. D.

1995-01-01

The search for evidence of life on Mars is a highly interdisciplinary enterprise which extends beyond the traditional life sciences. Mars conceivably had a pervasive ancient biosphere which may have persisted even to the present, but only in subsurface environments. Understanding the history of Mars' global environment, including its inventory of volatile elements, is a crucial part of the search strategy. Those deposits (minerals, sediments, etc.) which could have and retained a record of earlier biological activity must be identified and examined. While the importance of. seeking another biosphere has not diminished during the years since the Viking mission, the strategy for Mars exploration certainly has been modified by later discoveries. The Viking mission itself demonstrated that the present day surface environment of Mars is hostile to life as we know it. Thus, to search effectively for life on Mars, be it extant or extinct, we now must greatly improve our understanding of Mars the planet. Such an understanding will help us broaden our search beyond the Viking lander sites, both back in time to earlier epochs and elsewhere to other sites and beneath the surface. Exobiology involves much more than simply a search for extant life beyond Earth. It addresses the prospect of long-extinct biospheres and also the chemistry, organic and otherwise, which either led to life or which occurred on rocky planets that remained lifeless. Even a Mars without a biosphere would reveal much about life. How better to understand the origin and impact of a biosphere than to compare Earth with another similar but lifeless planet? Still, several relatively recent discoveries offer encouragement that a Martian biosphere indeed might have existed. The ancient Martian surface was extensively sculptured by volcanism and the activity of liquid water. Such observations invoke impressions of an ancient martian atmosphere and environment that resembled ancient Earth more than present-day Mars. Since Viking, we have learned that our own biosphere began prior to 3.5 billion years ago, during an early period when our solar system apparently was sustaining clement conditions on at least two of its planets. Also, we have found that microorganisms can survive, even flourish, in environments more extreme in temperature and water availability than had been previously recognized. The common ancestor of life on Earth probably was adapted to elevated temperatures, raising the possibility that hydrothermal systems played a central role in sustaining our early biosphere. If a biosphere ever arose on Mars, at least some of its constituents probably dwelled in the subsurface. Even today, conditions on Mars and Earth become more similar with increasing depth beneath their surfaces. For example, under the martian permafrost, the geothermal gradient very likely maintains liquid water in environments which resemble aquifers on Earth. Indigenous bacteria have recently been recovered from deep aquifers on Earth. Liquid groundwater very likely persisted throughout Mars' history. Thus, martian biota, if they ever existed, indeed might have survived in subsurface environments.

10 CFR 63.305 - Required characteristics of the reference biosphere.

Code of Federal Regulations, 2011 CFR

2011-01-01

....305 Section 63.305 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) DISPOSAL OF HIGH-LEVEL RADIOACTIVE... biosphere (other than climate), human biology, or increases or decreases of human knowledge or technology... factors remain constant as they are at the time of submission of the license application. (c) DOE must...

Bright Coherent Optical Waveforms from the Infrared to the Vacuum Ultraviolet for Manipulation and Detection of Molecules

DTIC Science & Technology

2012-12-13

Margaret Murnane. Invited talk, ITAMP Winter School on Atomic, Molecular and Optical Physics ( Biosphere 2, AZ, January 2012). McElvain Lecture...Molecular and Optical Physics ( Biosphere 2, AZ, January 2012). McElvain Lecture, University of Wisconsin Chemistry Department, February 2012. Seminar

A global biogeocenotical biosphere simulation

NASA Technical Reports Server (NTRS)

Moiseyev, N. N.

1980-01-01

This model of the D. Forrester type, constructed in differential equations, predicts the food and mineral supply for the factors biosphere population, depending on two socio-economic factors, until about the year 2500. If contemporary rates of natural resources utilization are maintained and there is no management of the environment, food resources will begin to limit human population growth after 2200, and mineral resources will after 2300. A decrease in the biosphere pollution, increase in effective agricultural production, and discovery of new energy sources may forestall or completely avert the onset of a crisis situation. Conservation measures, according to the model, are to a considerable extent realizable only if carried out simultaneously in both areas.

Biospheres and solar system exploration

NASA Technical Reports Server (NTRS)

Paine, Thomas O.

1990-01-01

The implications of biosphere technology is briefly examined. The exploration status and prospects of each world in the solar system is briefly reviewed, including the asteroid belt, the moon, and comets. Five program elements are listed as particularly critical for future interplanetary operations during the coming extraterrestrial century. They include the following: (1) a highway to Space (earth orbits); (2) Orbital Spaceports to support spacecraft assembly, storage, repair, maintenance, refueling, launch, and recovery; (3) a Bridge Between Worlds to transport cargo and crews to the moon and beyond to Mars; (4) Prospecting and Resource Utilization Systems to map and characterize the resources of planets, moons, and asteroids; and (5) Closed Ecology Biospheres. The progress in these five field is reviewed.

Mars Analog Rio Tinto Experiment (MARTE): 2003 Drilling Campaign to Search for a Subsurface Biosphere at Rio Tinto Spain

NASA Astrophysics Data System (ADS)

Stoker, C.; Dunagan, S.; Stevens, T.; Amils, R.; Gómez-Elvira, J.; Fernández, D.; Hall, J.; Lynch, K.; Cannon, H.; Zavaleta, J.; Glass, B.; Lemke, L.

2004-03-01

The results of an drilling experiment to search for a subsurface biosphere in a pyritic mineral deposit at Rio Tinto, Spain, are described. The experiment provides ground truth for a simulation of a Mars drilling mission to search for subsurface life.

76 FR 23649 - Endangered and Threatened Wildlife and Plants; 12-Month Finding on a Petition and Proposed Rule...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-27

... field research), the potential global population of free-ranging Morelet's crocodiles in 2004 was... will include monitoring and harvest of Morelet's crocodiles and hybrids for scientific research (CITES... Biosphere Reserve, Laguna de T[eacute]rminos Biosphere Reserve, Hampolol Wildlife Conservation and Research...

Putting the Deep Biosphere and Gas Hydrates on the Map

ERIC Educational Resources Information Center

Sikorski, Janelle J.; Briggs, Brandon R.

2016-01-01

Microbial processes in the deep biosphere affect marine sediments, such as the formation of gas hydrate deposits. Gas hydrate deposits offer a large source of natural gas with the potential to augment energy reserves and affect climate and seafloor stability. Despite the significant interdependence between life and geology in the ocean, coverage…

On the design of an interactive biosphere for the GLAS general circulation model

NASA Technical Reports Server (NTRS)

Mintz, Y.; Sellers, P. J.; Willmott, C. J.

1983-01-01

Improving the realism and accuracy of the GLAS general circulation model (by adding an interactive biosphere that will simulate the transfers of latent and sensible heat from land surface to atmosphere as functions of the atmospheric conditions and the morphology and physiology of the vegetation) is proposed.

The Large-Scale Biosphere-Atmosphere Experiment in Amazonia: Analyzing Regional Land Use Change Effects.

Treesearch

Michael Keller; Maria Assunção Silva-Dias; Daniel C. Nepstad; Meinrat O. Andreae

2004-01-01

The Large-Scale Biosphere-Atmosphere Experiment in Amazonia (LBA) is a multi-disciplinary, multinational scientific project led by Brazil. LBA researchers seek to understand Amazonia in its global context especially with regard to regional and global climate. Current development activities in Amazonia including deforestation, logging, cattle ranching, and agriculture...

The potential impacts of nutrient and CO2 variations on ecosystem oxidative ratio

USDA-ARS?s Scientific Manuscript database

A fraction of fossil fuel carbon dioxide (CO2) emissions are being taken up by the terrestrial biosphere and the oceans. One particularly effective way of determining the sizes of these terrestrial biosphere and ocean carbon sinks is based on the measurements of changes in atmospheric oxygen (O2) a...

International Co-ordinating Council of the Programme on Man and the Biosphere (MAB). Final Report.

ERIC Educational Resources Information Center

United Nations Educational, Scientific, and Cultural Organization, Paris (France).

Man and the Biosphere Program is an interdisciplinary program of research which emphasizes an ecological approach to the study of interrelationships between man and the environment. It is concerned with subjects of global or major regional significance which require international cooperation. This final report discusses areas in which…

Forest response to elevated CO2 is conserved across a broad range of productivity

Treesearch

R. Norby; E. DeLucia; B. Gielen; C. Calfapietra; C. Giardina; J. King; J. Ledford; H. McCarthy; D. Moore; R. Ceulemans; P. De Angelis; A. C. Finzi; D. F. Karnosky; M. E. Kubiske; M. Lukac; K. S. Pregitzer; G. E. Scarascia-Mugnozza; W. Schlesinger and R. Oren.

2005-01-01

Climate change predictions derived from coupled carbon-climate models are highly dependent on assumptions about feedbacks between the biosphere and atmosphere. One critical feedback occurs if C uptake by the biosphere increases in response to the fossil-fuel driven increase in atmospheric [CO2] ("CO2 fertilization...

Radiological risk assessment and biosphere modelling for radioactive waste disposal in Switzerland.

PubMed

Brennwald, M S; van Dorp, F

2009-12-01

Long-term safety assessments for geological disposal of radioactive waste in Switzerland involve the demonstration that the annual radiation dose to humans due to the potential release of radionuclides from the waste repository into the biosphere will not exceed the regulatory limit of 0.1 mSv. Here, we describe the simple but robust approach used by Nagra (Swiss National Cooperative for the Disposal of Radioactive Waste) to quantify the dose to humans as a result to time-dependent release of radionuclides from the geosphere into the biosphere. The model calculates the concentrations of radionuclides in different terrestrial and aquatic compartments of the surface environment. The fluxes of water and solids within the environment are the drivers for the exchange of radionuclides between these compartments. The calculated radionuclide concentrations in the biosphere are then used to estimate the radiation doses to humans due to various exposure paths (e.g. ingestion of radionuclides via drinking water and food, inhalation of radionuclides, external irradiation from radionuclides in soils). In this paper we also discuss recent new achievements and planned future work.

Atmospheric dynamics and bioregenerative technologies in a soil-based ecological life support system: Initial results from biosphere 2

NASA Astrophysics Data System (ADS)

Nelson, M.; Dempster, W.; Alvarez-Romo, N.; MacCallum, T.

1994-11-01

Biosphere 2 is the first man-made, soil-based, bioregenerative life support system to be developed and tested. The utilization and amendment of local space resources, e.g. martian soil or lunar regolith, for agricultural and other purposes will be necesary if we are to minimize the requirement for Earth materials in the creation of long-term off-planet bases and habitations. Several of the roles soil plays in Biosphere 2 are 1) for air purification 2) as a key component in created wetland systems to recycle human and animal wastes and 3) as nutrient base for a sustainable agricultural cropping program. Initial results from the Biosphere 2 closure experiment are presented. These include the accelerated cycling rates due to small reservoir sizes, strong diurnal and seasonal fluxes in atmospheric CO2, an unexpected and continuing decline in atmospheric oxygen, overall maintenance of low levels of trace gases, recycling of waste waters through biological regeneration systems, and operation of an agriculture designed to provide diverse and nutritionally adequate diets for the crew members.

Reconnecting to the biosphere.

PubMed

Folke, Carl; Jansson, Asa; Rockström, Johan; Olsson, Per; Carpenter, Stephen R; Chapin, F Stuart; Crépin, Anne-Sophie; Daily, Gretchen; Danell, Kjell; Ebbesson, Jonas; Elmqvist, Thomas; Galaz, Victor; Moberg, Fredrik; Nilsson, Måns; Osterblom, Henrik; Ostrom, Elinor; Persson, Asa; Peterson, Garry; Polasky, Stephen; Steffen, Will; Walker, Brian; Westley, Frances

2011-11-01

Humanity has emerged as a major force in the operation of the biosphere, with a significant imprint on the Earth System, challenging social-ecological resilience. This new situation calls for a fundamental shift in perspectives, world views, and institutions. Human development and progress must be reconnected to the capacity of the biosphere and essential ecosystem services to be sustained. Governance challenges include a highly interconnected and faster world, cascading social-ecological interactions and planetary boundaries that create vulnerabilities but also opportunities for social-ecological change and transformation. Tipping points and thresholds highlight the importance of understanding and managing resilience. New modes of flexible governance are emerging. A central challenge is to reconnect these efforts to the changing preconditions for societal development as active stewards of the Earth System. We suggest that the Millennium Development Goals need to be reframed in such a planetary stewardship context combined with a call for a new social contract on global sustainability. The ongoing mind shift in human relations with Earth and its boundaries provides exciting opportunities for societal development in collaboration with the biosphere--a global sustainability agenda for humanity.

Biological life-support systems for Mars mission.

PubMed

Gitelson, J I

1992-01-01

Mars mission like the Lunar base is the first venture to maintain human life beyond earth biosphere. So far, all manned space missions including the longest ones used stocked reserves and can not be considered egress from biosphere. Conventional path proposed by technology for Martian mission LSS is to use physical-chemical approaches proved by the experience of astronautics. But the problem of man living beyond the limits of the earth biosphere can be fundamentally solved by making a closed ecosystem for him. The choice optimum for a Mars mission LSS can be substantiated by comparing the merits and demerits of physical-chemical and biological principles without ruling out possible compromise between them. The work gives comparative analysis of ecological and physical-chemical principles for LSS. Taking into consideration universal significance of ecological problems with artificial LSS as a particular case of their solution, complexity and high cost of large-scale experiments with manned LSS, it would be expedient for these works to have the status of an International Program open to be joined. A program of making artificial biospheres based on preceding experience and analysis of current situation is proposed.

Comparing the effectiveness of monetary versus moral motives in environmental campaigning

NASA Astrophysics Data System (ADS)

Bolderdijk, J. W.; Steg, L.; Geller, E. S.; Lehman, P. K.; Postmes, T.

2013-04-01

Environmental campaigns often promote energy conservation by appealing to economic (for example, lower electricity bills) rather than biospheric concerns (for example, reduced carbon emissions), assuming that people are primarily motivated by economic self-interest. However, people also care about maintaining a favourable view of themselves (they want to maintain a `positive self-concept'), and may prefer to see themselves as `green' rather than `greedy'. Consequently, people may find economic appeals less attractive than biospheric appeals. Across two studies, participants indicated feeling better about biospheric (`Want to protect the environment? Check your car's tire pressure') than economic (`Want to save money? Check your car's tire pressure') tyre-check appeals. In a field experiment, we found that an economic tyre-check appeal (`Do you care about your finances? Get a free tire check') elicited significantly less compliance than parallel biospheric and neutral appeals. Together, these studies discredit the conventional wisdom that appealing to economic self-interest is the best way to secure behaviour change. At least in some cases, our studies suggest, this strategy is not effective.

Pharmaceutical Residues Affecting the UNESCO Biosphere Reserve Kristianstads Vattenrike Wetlands: Sources and Sinks.

PubMed

Björklund, Erland; Svahn, Ola; Bak, Søren; Bekoe, Samuel Oppong; Hansen, Martin

2016-10-01

This study is the first to investigate the pharmaceutical burden from point sources affecting the UNESCO Biosphere Reserve Kristianstads Vattenrike, Sweden. The investigated Biosphere Reserve is a >1000 km(2) wetland system with inflows from lakes, rivers, leachate from landfill, and wastewater-treatment plants (WWTPs). We analysed influent and treated wastewater, leachate water, lake, river, and wetland water alongside sediment for six model pharmaceuticals. The two WWTPs investigated released pharmaceutical residues at levels close to those previously observed in Swedish monitoring exercises. Compound-dependent WWTP removal efficiencies ranging from 12 to 100 % for bendroflumethiazide, oxazepam, atenolol, carbamazepine, and diclofenac were observed. Surface-water concentrations in the most affected lake were ≥100 ng/L for the various pharmaceuticals with atenolol showing the highest levels (>300 ng/L). A small risk assessment showed that adverse single-substance toxicity on aquatic organisms within the UNESCO Biosphere Reserve is unlikely. However, the effects of combinations of a large number of known and unknown pharmaceuticals, metals, and nutrients are still unknown.

The Effects of Recreation Experience, Environmental Attitude, and Biospheric Value on the Environmentally Responsible Behavior of Nature-Based Tourists

NASA Astrophysics Data System (ADS)

Lee, Tsung Hung; Jan, Fen-Hauh

2015-07-01

The scientific understanding of the recreation experience and the environmentally responsible behavior of nature-based tourists is limited. This study examines the relationship among the recreation experience, environmental attitude, biospheric value, and the general and site-specific environmentally responsible behavior of nature-based tourists in Taomi, Liuqiu Island, and Aowanda and Najenshan in Taiwan. A total of 1342 usable questionnaires were collected for this study. The empirical results indicate that the recreation experience influences biospheric value and environmental attitude; subsequently, it then indirectly influences the general and site-specific environmentally responsible behavior of nature-based tourists. Our theoretical behavioral model elucidates previously proposed but unexamined behavioral models among nature-based tourists, and it offers a theoretical framework for researchers, decision makers, managers, and tourists in the field of nature-based tourism. We conclude that when an individual participates in nature-based tourism as described here, these recreation experiences strengthen their environmental attitude and biospheric value, and consequently increase their engagement in both general and site-specific environmentally responsible behaviors.

The effects of recreation experience, environmental attitude, and biospheric value on the environmentally responsible behavior of nature-based tourists.

PubMed

Lee, Tsung Hung; Jan, Fen-Hauh

2015-07-01

The scientific understanding of the recreation experience and the environmentally responsible behavior of nature-based tourists is limited. This study examines the relationship among the recreation experience, environmental attitude, biospheric value, and the general and site-specific environmentally responsible behavior of nature-based tourists in Taomi, Liuqiu Island, and Aowanda and Najenshan in Taiwan. A total of 1342 usable questionnaires were collected for this study. The empirical results indicate that the recreation experience influences biospheric value and environmental attitude; subsequently, it then indirectly influences the general and site-specific environmentally responsible behavior of nature-based tourists. Our theoretical behavioral model elucidates previously proposed but unexamined behavioral models among nature-based tourists, and it offers a theoretical framework for researchers, decision makers, managers, and tourists in the field of nature-based tourism. We conclude that when an individual participates in nature-based tourism as described here, these recreation experiences strengthen their environmental attitude and biospheric value, and consequently increase their engagement in both general and site-specific environmentally responsible behaviors.

Harvesting the biosphere: the human impact.

PubMed

Smil, Vaclav

2011-01-01

The human species has evolved to dominate the biosphere: global anthropomass is now an order of magnitude greater than the mass of all wild terrestrial mammals. As a result, our dependence on harvesting the products of photosynthesis for food, animal feed, raw materials, and energy has grown to make substantial global impacts. During the past two millennia these harvests, and changes of land use due to deforestation and conversions of grasslands and wetlands, have reduced the stock of global terrestrial plant mass by as much as 45 percent, with the twentieth-century reduction amounting to more than 15 percent. Current annual harvests of phytomass have been a significant share of the global net primary productivity (NPP, the total amount of new plant tissues created by photosynthesis). Some studies put the human appropriation of NPP (the ratio of these two variables) as high as 40 percent but the measure itself is problematic. Future population growth and improved quality of life will result in additional claims on the biosphere, but options to accommodate these demands exist without severely compromising the irreplaceable biospheric services.

Phylogenetic Characterization of Fecal Microbial Communities of Dogs Fed Diets with or without Supplemental Dietary Fiber Using 454 Pyrosequencing

PubMed Central

Middelbos, Ingmar S.; Vester Boler, Brittany M.; Qu, Ani; White, Bryan A.; Swanson, Kelly S.; Fahey, George C.

2010-01-01

Background Dogs suffer from many of the same maladies as humans that may be affected by the gut microbiome, but knowledge of the canine microbiome is incomplete. This work aimed to use 16S rDNA tag pyrosequencing to phylogenetically characterize hindgut microbiome in dogs and determine how consumption of dietary fiber affects community structure. Principal Findings Six healthy adult dogs were used in a crossover design. A control diet without supplemental fiber and a beet pulp-supplemented (7.5%) diet were fed. Fecal DNA was extracted and the V3 hypervariable region of the microbial 16S rDNA gene amplified using primers suitable for 454-pyrosequencing. Microbial diversity was assessed on random 2000-sequence subsamples of individual and pooled DNA samples by diet. Our dataset comprised 77,771 reads with an average length of 141 nt. Individual samples contained approximately 129 OTU, with Fusobacteria (23 – 40% of reads), Firmicutes (14 – 28% of reads) and Bacteroidetes (31 – 34% of reads) being co-dominant phyla. Feeding dietary fiber generally decreased Fusobacteria and increased Firmicutes, but these changes were not equally apparent in all dogs. UniFrac analysis revealed that structure of the gut microbiome was affected by diet and Firmicutes appeared to play a strong role in by-diet clustering. Conclusions Our data suggest three co-dominant bacterial phyla in the canine hindgut. Furthermore, a relatively small amount of dietary fiber changed the structure of the gut microbiome detectably. Our data are among the first to characterize the healthy canine gut microbiome using pyrosequencing and provide a basis for studies focused on devising dietary interventions for microbiome-associated diseases. PMID:20339542

A pyrosequencing assay for the quantitative methylation analysis of the PCDHB gene cluster, the major factor in neuroblastoma methylator phenotype.

PubMed

Banelli, Barbara; Brigati, Claudio; Di Vinci, Angela; Casciano, Ida; Forlani, Alessandra; Borzì, Luana; Allemanni, Giorgio; Romani, Massimo

2012-03-01

Epigenetic alterations are hallmarks of cancer and powerful biomarkers, whose clinical utilization is made difficult by the absence of standardization and of common methods of data interpretation. The coordinate methylation of many loci in cancer is defined as 'CpG island methylator phenotype' (CIMP) and identifies clinically distinct groups of patients. In neuroblastoma (NB), CIMP is defined by a methylation signature, which includes different loci, but its predictive power on outcome is entirely recapitulated by the PCDHB cluster only. We have developed a robust and cost-effective pyrosequencing-based assay that could facilitate the clinical application of CIMP in NB. This assay permits the unbiased simultaneous amplification and sequencing of 17 out of 19 genes of the PCDHB cluster for quantitative methylation analysis, taking into account all the sequence variations. As some of these variations were at CpG doublets, we bypassed the data interpretation conducted by the methylation analysis software to assign the corrected methylation value at these sites. The final result of the assay is the mean methylation level of 17 gene fragments in the protocadherin B cluster (PCDHB) cluster. We have utilized this assay to compare the methylation levels of the PCDHB cluster between high-risk and very low-risk NB patients, confirming the predictive value of CIMP. Our results demonstrate that the pyrosequencing-based assay herein described is a powerful instrument for the analysis of this gene cluster that may simplify the data comparison between different laboratories and, in perspective, could facilitate its clinical application. Furthermore, our results demonstrate that, in principle, pyrosequencing can be efficiently utilized for the methylation analysis of gene clusters with high internal homologies.

Clinical Implications of Quantitative JAK2 V617F Analysis using Droplet Digital PCR in Myeloproliferative Neoplasms

PubMed Central

Lee, Eunyoung; Lee, Kyoung Joo; Park, Hyein; Chung, Jin Young; Lee, Mi-Na; Chang, Myung Hee; Yoo, Jongha; Lee, Hyewon

2018-01-01

Background JAK2 V617F is the most common mutation in myeloproliferative neoplasms (MPNs) and is a major diagnostic criterion. Mutation quantification is useful for classifying patients with MPN into subgroups and for prognostic prediction. Droplet digital PCR (ddPCR) can provide accurate and reproducible quantitative analysis of DNA. This study was designed to verify the correlation of ddPCR with pyrosequencing results in the diagnosis of MPN and to investigate clinical implications of the mutational burden. Methods Peripheral blood or bone marrow samples were obtained from 56 patients newly diagnosed with MPN or previously diagnosed with MPN but not yet indicated for JAK2 inhibitor treatment between 2012 and 2016. The JAK2 V617F mutation was detected by pyrosequencing as a diagnostic work-up. The same samples were used for ddPCR to determine the correlation between assays and establish a detection sensitivity cut-off. Clinical and hematologic aspects were reviewed. Results Forty-two (75%) and 46 (82.1%) patients were positive for JAK2 V617F by pyrosequencing and ddPCR, respectively. The mean mutated allele frequency at diagnosis was 37.5±30.1% and was 40.7±31.2% with ddPCR, representing a strong correlation (r=0.9712, P<0.001). Follow-up samples were available for 12 patients, including eight that were JAK2 V617F-positive. Of these, mutational burden reduction after treatment was observed in six patients (75%), consistent with trends of hematologic improvement. Conclusions Quantitative analysis of the JAK2 V617F mutation using ddPCR was highly correlated with pyrosequencing data and may reflect the clinical response to treatment. PMID:29214759

A molecular gram stain using broad range PCR and pyrosequencing technology: a potentially useful tool for diagnosing orthopaedic infections.

PubMed

Kobayashi, Naomi; Bauer, Thomas W; Togawa, Daisuke; Lieberman, Isador H; Sakai, Hiroshige; Fujishiro, Takaaki; Tuohy, Marion J; Procop, Gary W

2005-06-01

The bacteria associated with orthopaedic infections are usually common gram-positive and gram-negative bacteria. This fundamental grouping of bacteria is a necessary first step in the selection of appropriate antibiotics. Since polymerase chain reaction (PCR) is more rapid and may be more sensitive than culture, we developed a postamplification pyrosequencing method to subcategorize bacteria based on a few nucleotide polymorphisms in the 16S rRNA gene. We validated this method using well-characterized strains of bacteria and applied it to specimens from spinal surgery cases with suspected infections. Lysates of 114 bacteria including 75 species were created following standard cultivation to obtain DNA. The DNA was amplified by a broad-range real-time PCR. The amplicons were evaluated by pyrosequencing and were classified as gram-positive, gram-negative, or acid-fast bacilli based on the first three to five nucleotides sequenced. In addition, clinical cases of suspected infection were obtained from spinal surgery. The results of the "molecular Gram stain" were compared with the results of traditional Gram stain and culture. The lysates of 107 (93.9%) of the bacteria extracts tested were appropriately categorized as gram-positive and gram-negative or as acid-fast bacilli on the basis of this assay. The sensitivity and specificity of this assay were 100% and 97.4% for gram-positive and 88.3% and 100% for gram-negative isolates. All of the five clinical samples were appropriately categorized as containing gram-positive or gram-negative bacteria with this assay. This study demonstrates that high sensitivity and specificity of a molecular gram stain may be achieved using broad-range real-time PCR and pyrosequencing.

Putting the Deep Biosphere on the Map for Oceanography Courses: Gas Hydrates As a Case Study for the Deep Biosphere

NASA Astrophysics Data System (ADS)

Sikorski, J. J.; Briggs, B. R.

2014-12-01

The ocean is essential for life on our planet. It covers 71% of the Earth's surface, is the source of the water we drink, the air we breathe, and the food we eat. Yet, the exponential growth in human population is putting the ocean and thus life on our planet at risk. However, based on student evaluations from our introductory oceanography course it is clear that our students have deficiencies in ocean literacy that impact their ability to recognize that the ocean and humans are inextricably connected. Furthermore, life present in deep subsurface marine environments is also interconnected to the study of the ocean, yet the deep biosphere is not typically covered in undergraduate oceanography courses. In an effort to improve student ocean literacy we developed an instructional module on the deep biosphere focused on gas hydrate deposits. Specifically, our module utilizes Google Earth and cutting edge research about microbial life in the ocean to support three inquiry-based activities that each explore different facets of gas hydrates (i.e. environmental controls, biologic controls, and societal implications). The relevant nature of the proposed module also makes it possible for instructors of introductory geology courses to modify module components to discuss related topics, such as climate, energy, and geologic hazards. This work, which will be available online as a free download, is a solid contribution toward increasing the available teaching resources focused on the deep biosphere for geoscience educators.

Aeolian Processes and the Biosphere

NASA Astrophysics Data System (ADS)

Ravi, Sujith; D'Odorico, Paolo; Breshears, David D.; Field, Jason P.; Goudie, Andrew S.; Huxman, Travis E.; Li, Junran; Okin, Gregory S.; Swap, Robert J.; Thomas, Andrew D.; Van Pelt, Scott; Whicker, Jeffrey J.; Zobeck, Ted M.

2011-08-01

Aeolian processes affect the biosphere in a wide variety of contexts, including landform evolution, biogeochemical cycles, regional climate, human health, and desertification. Collectively, research on aeolian processes and the biosphere is developing rapidly in many diverse and specialized areas, but integration of these recent advances is needed to better address management issues and to set future research priorities. Here we review recent literature on aeolian processes and their interactions with the biosphere, focusing on (1) geography of dust emissions, (2) impacts, interactions, and feedbacks, (3) drivers of dust emissions, and (4) methodological approaches. Geographically, dust emissions are highly spatially variable but also provide connectivity at global scales between sources and effects, with “hot spots” being of particular concern. Recent research reveals that aeolian processes have impacts, interactions, and feedbacks at a variety of scales, including large-scale dust transport and global biogeochemical cycles, climate mediated interactions between atmospheric dust and ecosystems, impacts on human health, impacts on agriculture, and interactions between aeolian processes and dryland vegetation. Aeolian dust emissions are driven largely by, in addition to climate, a combination of soil properties, soil moisture, vegetation and roughness, biological and physical crusts, and disturbances. Aeolian research methods span laboratory and field techniques, modeling, and remote sensing. Together these integrated perspectives on aeolian processes and the biosphere provide insights into management options and aid in identifying research priorities, both of which are increasingly important given that global climate models predict an increase in aridity in many dryland systems of the world.

A model of the CO2 exchanges between biosphere and atmosphere in the tundra

NASA Technical Reports Server (NTRS)

Labgaa, Rachid R.; Gautier, Catherine

1992-01-01

A physical model of the soil thermal regime in a permafrost terrain has been developed and validated with soil temperature measurements at Barrow, Alaska. The model calculates daily soil temperatures as a function of depth and average moisture contents of the organic and mineral layers using a set of five climatic variables, i.e., air temperature, precipitation, cloudiness, wind speed, and relative humidity. The model is not only designed to study the impact of climate change on the soil temperature and moisture regime, but also to provide the input to a decomposition and net primary production model. In this context, it is well known that CO2 exchanges between the terrestrial biosphere and the atmosphere are driven by soil temperature through decomposition of soil organic matter and root respiration. However, in tundra ecosystems, net CO2 exchange is extremely sensitive to soil moisture content; therefore it is necessary to predict variations in soil moisture in order to assess the impact of climate change on carbon fluxes. To this end, the present model includes the representation of the soil moisture response to changes in climatic conditions. The results presented in the foregoing demonstrate that large errors in soil temperature and permafrost depth estimates arise from neglecting the dependence of the soil thermal regime on soil moisture contents. Permafrost terrain is an example of a situation where soil moisture and temperature are particularly interrelated: drainage conditions improve when the depth of the permafrost increases; a decrease in soil moisture content leads to a decrease in the latent heat required for the phase transition so that the heat penetrates faster and deeper, and the maximum depth of thaw increases; and as excepted, soil thermal coefficients increase with moisture.

Bayesian optimization of the Community Land Model simulated biosphere-atmosphere exchange using CO 2 observations from a dense tower network and aircraft campaigns over Oregon

DOE PAGES

Schmidt, Andres; Law, Beverly E.; Göckede, Mathias; ...

2016-09-15

Here, the vast forests and natural areas of the Pacific Northwest comprise one of the most productive ecosystems in the northern hemisphere. The heterogeneous landscape of Oregon poses a particular challenge to ecosystem models. We present a framework using a scaling factor Bayesian inversion to improve the modeled atmosphere-biosphere exchange of carbon dioxide. Observations from 5 CO/CO 2 towers, eddy covariance towers, and airborne campaigns were used to constrain the Community Land Model CLM4.5 simulated terrestrial CO 2 exchange at a high spatial and temporal resolution (1/24°, 3-hourly). To balance aggregation errors and the degrees of freedom in the inversemore » modeling system, we applied an unsupervised clustering approach for the spatial structuring of our model domain. Data from flight campaigns were used to quantify the uncertainty introduced by the Lagrangian particle dispersion model that was applied for the inversions. The average annual statewide net ecosystem productivity (NEP) was increased by 32% to 29.7 TgC per year by assimilating the tropospheric mixing ratio data. The associated uncertainty was decreased by 28.4% to 29%, on average over the entire Oregon model domain with the lowest uncertainties of 11% in western Oregon. The largest differences between posterior and prior CO 2 fluxes were found for the Coast Range ecoregion of Oregon that also exhibits the highest availability of atmospheric observations and associated footprints. In this area, covered by highly productive Douglas-fir forest, the differences between the prior and posterior estimate of NEP averaged 3.84 TgC per year during the study period from 2012 through 2014.« less

Bayesian optimization of the Community Land Model simulated biosphere-atmosphere exchange using CO 2 observations from a dense tower network and aircraft campaigns over Oregon

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schmidt, Andres; Law, Beverly E.; Göckede, Mathias

Here, the vast forests and natural areas of the Pacific Northwest comprise one of the most productive ecosystems in the northern hemisphere. The heterogeneous landscape of Oregon poses a particular challenge to ecosystem models. We present a framework using a scaling factor Bayesian inversion to improve the modeled atmosphere-biosphere exchange of carbon dioxide. Observations from 5 CO/CO 2 towers, eddy covariance towers, and airborne campaigns were used to constrain the Community Land Model CLM4.5 simulated terrestrial CO 2 exchange at a high spatial and temporal resolution (1/24°, 3-hourly). To balance aggregation errors and the degrees of freedom in the inversemore » modeling system, we applied an unsupervised clustering approach for the spatial structuring of our model domain. Data from flight campaigns were used to quantify the uncertainty introduced by the Lagrangian particle dispersion model that was applied for the inversions. The average annual statewide net ecosystem productivity (NEP) was increased by 32% to 29.7 TgC per year by assimilating the tropospheric mixing ratio data. The associated uncertainty was decreased by 28.4% to 29%, on average over the entire Oregon model domain with the lowest uncertainties of 11% in western Oregon. The largest differences between posterior and prior CO 2 fluxes were found for the Coast Range ecoregion of Oregon that also exhibits the highest availability of atmospheric observations and associated footprints. In this area, covered by highly productive Douglas-fir forest, the differences between the prior and posterior estimate of NEP averaged 3.84 TgC per year during the study period from 2012 through 2014.« less

Exploration of a Subsurface Biosphere in a Volcanic Massive Sulfide: Results of the Mars Analog Rio Tinto Drilling Experiment

NASA Astrophysics Data System (ADS)

Stoker, C. R.; Stevens, T.; Amils, R.; Fernandez, D.

2005-12-01

Biological systems on Earth require three key ingredients-- liquid water, an energy source, and a carbon source, that are found in very few extraterrestrial environments. Previous examples of independent subsurface ecosystems have been found only in basalt aquifers. Such lithotrophic microbial ecosystems (LME) have been proposed as models for steps in the early evolution of Earth's biosphere and for potential biospheres on other planets where the surface is uninhabitable, such as Mars and Europa.. The Mars Analog Rio Tinto Experiment (MARTE) has searched in a volcanic massive sulfide deposit in Rio Tinto Spain for a subsurface biosphere capable of living without sunlight or oxygen and found a subsurface ecosystem driven by the weathering of the massive sulfide deposit (VMS) in which the rock matrix provides sufficient resources to support microbial metabolism, including the vigorous production of H2 by water-rock interactions. Microbial production of methane and sulfate occurred in the sulfide orebody and microbial production of methane and hydrogen sulfide continued in an anoxic plume downgradient from the sulfide ore. Organic carbon concentrations in the parent rock were too low to support microbes. The Rio Tinto system thus represents a new type of subsurface ecosystem with strong relevance for exobiological studies. Commercial drilling was used to reach the aquifer system at 100 m depth and conventional laboratory techniques were used to identify and characterize the biosphere. Then, the life search strategy that led to successful identification of this biosphere was applied to the development of a robotic drilling, core handling, inspection, subsampling, and life detection system built on a prototype planetary lander that was deployed in Rio Tinto Spain in September 2005 to test the capability of a robotic drilling system to search for subsurface life. A remote science team directed the simulation and analyzed the data from the MARTE robotic drill. The results of this experiment have important implications for the strategy for searching for life on Mars.

“Modular Biospheres” New testbed platforms for public environmental education and research

NASA Astrophysics Data System (ADS)

Nelson, M.; Dempster, W. F.; Allen, J. P.

This paper will review the potential of a relatively new type of testbed platform for environmental education and research because of the unique advantages resulting from their material closure and separation from the outside environment. These facilities which we term "modular biospheres", have emerged from research centered on space life support research but offer a wider range of application. Examples of this type of facility include the Bios-3 facility in Russia, the Japanese CEEF (Closed Ecological Experiment Facility), the NASA Kennedy Space Center Breadboard facility, the Biosphere 2 Test Module and the Laboratory Biosphere. Modular biosphere facilities offer unique research and public real-time science education opportunities. Ecosystem behavior can be studied since initial state conditions can be precisely specified and tracked over different ranges of time. With material closure (apart from very small air exchange rate which can be determined), biogeochemical cycles between soil and soil microorganisms, water, plants, and atmosphere can be studied in detail. Such studies offer a major advance from studies conducted with phytotrons which because of their small size, limit the number of organisms to a very small number, and which crucially do not have a high degree of atmospheric, water and overall material closure. Modular biospheres take advantage of the unique properties of closure, as representing a distinct system "metabolism" and therefore are essentially a "mini-world". Though relatively large in comparison with most phytotrons and ecological microcosms, which are now standard research and educational tools, modular biospheres are small enough that they can be economically reconfigured to reflect a changing research agenda. Some design elements include lighting via electric lights and/or sunlight, hydroponic or soil substrate for plants, opaque or glazed structures, and variable volume chambers or other methods to handle atmospheric pressure differences between the facility and the outside environment.

A comparison between the example reference biosphere model ERB 2B and a process-based model: simulation of a natural release scenario.

PubMed

Almahayni, T

2014-12-01

The BIOMASS methodology was developed with the objective of constructing defensible assessment biospheres for assessing potential radiological impacts of radioactive waste repositories. To this end, a set of Example Reference Biospheres were developed to demonstrate the use of the methodology and to provide an international point of reference. In this paper, the performance of the Example Reference Biosphere model ERB 2B associated with the natural release scenario, discharge of contaminated groundwater to the surface environment, was evaluated by comparing its long-term projections of radionuclide dynamics and distribution in a soil-plant system to those of a process-based, transient advection-dispersion model (AD). The models were parametrised with data characteristic of a typical rainfed winter wheat crop grown on a sandy loam soil under temperate climate conditions. Three safety-relevant radionuclides, (99)Tc, (129)I and (237)Np with different degree of sorption were selected for the study. Although the models were driven by the same hydraulic (soil moisture content and water fluxes) and radiological (Kds) input data, their projections were remarkably different. On one hand, both models were able to capture short and long-term variation in activity concentration in the subsoil compartment. On the other hand, the Reference Biosphere model did not project any radionuclide accumulation in the topsoil and crop compartments. This behaviour would underestimate the radiological exposure under natural release scenarios. The results highlight the potential role deep roots play in soil-to-plant transfer under a natural release scenario where radionuclides are released into the subsoil. When considering the relative activity and root depth profiles within the soil column, much of the radioactivity was taken up into the crop from the subsoil compartment. Further improvements were suggested to address the limitations of the Reference Biosphere model presented in this paper. Copyright © 2014 Elsevier Ltd. All rights reserved.

Clustering XCO2 temporal change to assess CO2 exchanging strength of biosphere-atmosphere with GOSAT observations

NASA Astrophysics Data System (ADS)

He, Zhonghua; Lei, Liping; Bie, Nian; Yang, Shaoyuan; Wu, Changjiang; Zeng, Zhao-Cheng

2017-04-01

The temporal change of atmospheric carbon dioxide (CO2) concentration, greatly related to the local activities of CO2 uptake and emission, including biospheric exchange and anthropogenic emission, is one of important information for regions identification of carbon source and sink. Satellite observations of CO2 has been used for detecting the change of CO2 concentration for a long time. In this study, we used the grid data of column-averaged CO2 dry air mole fraction (XCO2) with the spatial resolution of 1 degree and the temporal resolution of 3 days from 1 June 2009 to 31 May 2014 over the land area of 30° - 60° N to implement a clustering of temporal changing characteristics for the Greenhouse Gases Observing Satellite (GOSAT) XCO2 retrievals. Grid data is derived using the gap filling method of spatio-temporal geostatistics. The clustering method is one adjusted K-mean for the gap existed time-series data. As a result, types and number of clusters are specified based on the temporal characteristic of XCO2 by using the optimal clustering parameters. The biospheric absorption and surface emission of atmospheric CO2 is discussed through the analysis of the different yearly increase and seasonal amplitude of XCO2 each cluster combined with correlation analysis with vegetation index from the Moderate-resolution Imaging Spectroradiometer (MODIS) and fossil fuel CO2 emission data from Open-source Data Inventory for Anthropogenic CO2 (Odiac). Regions of strong or weak biosphere-atmosphere exchange, or significant disturbance from anthropogenic activities can be identified. In conclusion, gap filled XCO2 from satellite observations can help us to take an analysis of atmospheric CO2, results of the coupled biosphere-atmosphere, by their spatio-temporal characteristics as well as the relationship with the other remote sensing parameters e.g. MODIS related with biospheric photosynthetic or respiration activities.

Sequential optimization of a terrestrial biosphere model constrained by multiple satellite based products

NASA Astrophysics Data System (ADS)

Ichii, K.; Kondo, M.; Wang, W.; Hashimoto, H.; Nemani, R. R.

2012-12-01

Various satellite-based spatial products such as evapotranspiration (ET) and gross primary productivity (GPP) are now produced by integration of ground and satellite observations. Effective use of these multiple satellite-based products in terrestrial biosphere models is an important step toward better understanding of terrestrial carbon and water cycles. However, due to the complexity of terrestrial biosphere models with large number of model parameters, the application of these spatial data sets in terrestrial biosphere models is difficult. In this study, we established an effective but simple framework to refine a terrestrial biosphere model, Biome-BGC, using multiple satellite-based products as constraints. We tested the framework in the monsoon Asia region covered by AsiaFlux observations. The framework is based on the hierarchical analysis (Wang et al. 2009) with model parameter optimization constrained by satellite-based spatial data. The Biome-BGC model is separated into several tiers to minimize the freedom of model parameter selections and maximize the independency from the whole model. For example, the snow sub-model is first optimized using MODIS snow cover product, followed by soil water sub-model optimized by satellite-based ET (estimated by an empirical upscaling method; Support Vector Regression (SVR) method; Yang et al. 2007), photosynthesis model optimized by satellite-based GPP (based on SVR method), and respiration and residual carbon cycle models optimized by biomass data. As a result of initial assessment, we found that most of default sub-models (e.g. snow, water cycle and carbon cycle) showed large deviations from remote sensing observations. However, these biases were removed by applying the proposed framework. For example, gross primary productivities were initially underestimated in boreal and temperate forest and overestimated in tropical forests. However, the parameter optimization scheme successfully reduced these biases. Our analysis shows that terrestrial carbon and water cycle simulations in monsoon Asia were greatly improved, and the use of multiple satellite observations with this framework is an effective way for improving terrestrial biosphere models.

Evaluation of simulated biospheric carbon dioxide fluxes and atmospheric concentrations using global in situ observations

NASA Astrophysics Data System (ADS)

Philip, S.; Johnson, M. S.; Potter, C. S.; Genovese, V. B.

2016-12-01

Atmospheric mixing ratios of carbon dioxide (CO2) are largely controlled by anthropogenic emission sources and biospheric sources/sinks. Global biospheric fluxes of CO2 are controlled by complex processes facilitating the exchange of carbon between terrestrial ecosystems and the atmosphere. These processes which play a key role in these terrestrial ecosystem-atmosphere carbon exchanges are currently not fully understood, resulting in large uncertainties in the quantification of biospheric CO2 fluxes. Current models with these inherent deficiencies have difficulties simulating the global carbon cycle with high accuracy. We are developing a new modeling platform, GEOS-Chem-CASA by integrating the year-specific NASA-CASA (National Aeronautics and Space Administration - Carnegie Ames Stanford Approach) biosphere model with the GEOS-Chem (Goddard Earth Observation System-Chemistry) chemical transport model to improve the simulation of atmosphere-terrestrial ecosystem carbon exchange. We use NASA-CASA to explicitly represent the exchange of CO2 between terrestrial ecosystem and atmosphere by replacing the baseline GEOS-Chem land net CO2 flux and forest biomass burning CO2 emissions. We will present the estimation and evaluation of these "bottom-up" land CO2 fluxes, simulated atmospheric mixing ratios, and forest disturbance changes over the last decade. In addition, we will present our initial comparison of atmospheric column-mean dry air mole fraction of CO2 predicted by the model and those retrieved from NASA's OCO-2 (Orbiting Carbon Observatory-2) satellite instrument and model-predicted surface CO2 mixing ratios with global in situ observations. This evaluation is the first step necessary for our future work planned to constrain the estimates of biospheric carbon fluxes through "top-down" inverse modeling, which will improve our understanding of the processes controlling atmosphere-terrestrial ecosystem greenhouse gas exchanges, especially over regions which lack in situ observations.

B33C-0612: Evaluation of Simulated Biospheric Carbon Dioxide Fluxes and Atmospheric Concentrations Using Global in Situ Observations

NASA Technical Reports Server (NTRS)

Philip, Sajeev; Johnson, Matthew S.; Potter, Christopher S.; Genovese, Vanessa

2016-01-01

Atmospheric mixing ratios of carbon dioxide (CO2) are largely controlled by anthropogenic emission sources and biospheric sources/sinks. Global biospheric fluxes of CO2 are controlled by complex processes facilitating the exchange of carbon between terrestrial ecosystems and the atmosphere. These processes which play a key role in these terrestrial ecosystem-atmosphere carbon exchanges are currently not fully understood, resulting in large uncertainties in the quantification of biospheric CO2 fluxes. Current models with these inherent deficiencies have difficulties simulating the global carbon cycle with high accuracy. We are developing a new modeling platform, GEOS-Chem-CASA by integrating the year-specific NASA-CASA (National Aeronautics and Space Administration - Carnegie Ames Stanford Approach) biosphere model with the GEOS-Chem (Goddard Earth Observation System-Chemistry) chemical transport model to improve the simulation of atmosphere-terrestrial ecosystem carbon exchange. We use NASA-CASA to explicitly represent the exchange of CO2 between terrestrial ecosystem and atmosphere by replacing the baseline GEOS-Chem land net CO2 flux and forest biomass burning CO2 emissions. We will present the estimation and evaluation of these "bottom-up" land CO2 fluxes, simulated atmospheric mixing ratios, and forest disturbance changes over the last decade. In addition, we will present our initial comparison of atmospheric column-mean dry air mole fraction of CO2 predicted by the model and those retrieved from NASA's OCO-2 (Orbiting Carbon Observatory-2) satellite instrument and model-predicted surface CO2 mixing ratios with global in situ observations. This evaluation is the first step necessary for our future work planned to constrain the estimates of biospheric carbon fluxes through "top-down" inverse modeling, which will improve our understanding of the processes controlling atmosphere-terrestrial ecosystem greenhouse gas exchanges, especially over regions which lack in situ observations.

[Microbes on the edge of global biosphere].

PubMed

Naganuma, T

2000-12-01

The search for life on the edge of global biosphere is a frontier to bridge conventional bio/ecology and exo/astrobiology. This communication reviews the foci of microbiological studies on the inhabitants of the selected "edges", i.e., deep-sea, deep subsurface and Antarctic habitats. The deep-sea is characterized as the no-light (non-photosynthetic) habitat, and the primary production is mostly due to the chemosynthetic autotrophy at the hydrothermal vents and methane-rich seeps. Formation of the chemosynthesis-dependent animal communities in the deep leads to the idea that such communities may be found in "ocean" of the Jovian satellite, Europa. The oxygen minimal layer (OML) in mid-water provides another field of deep-sea research. Modern OML is a relatively thin layer, found between the water depth of 200 and 1000 m, but was much thicker during the periods of oceanic anoxia events (OAEs) in the past. The history of oceanic biosphere is regarded as the cycle of OAE and non-OAE periods, and the remnants of the past OAEs may be seen in the modem OML. Anoxic (no-O2) condition is also characteristic of deep subsurface biosphere. Microorganisms in deep subsurface biosphere exploit every available oxidant, or terminal electron acceptor (TEA), for anaerobic respiration. Sulfate, nitrate, iron (III) and CO2 are the representative TEAs in the deep subsurface. Subsurface of hydrothermal vents, or sub-vent biosphere, may house brine (high salt) habitats and halophilic microorganisms. Some sub-vent halophiles were phylogenetically closely similar to the ones found in the Antarctic habitats which are extremely dry by the liophilizing climate. Below the 3000-4000 m-thick glacier on Antarctica, there have been >70 lakes with liquid water located. One of such sub-glacial lakes, Lake Vostok, has been a target of "life in extreme environments" and is about to be drill-penetrated for microbiological studies. These 'microbiological platforms' will provide new knowledge about the diversity and potential of the Earth's life and facilitate the capability of astrobiologial exploration.

The past, present and future supernova threat to Earth's biosphere

NASA Astrophysics Data System (ADS)

Beech, Martin

2011-12-01

A brief review of the threat posed to Earth's biosphere via near-by supernova detonations is presented. The expected radiation dosage, cosmic ray flux and expanding blast wave collision effects are considered, and it is argued that a typical supernova must be closer than ˜10-pc before any appreciable and potentially harmful atmosphere/biosphere effects are likely to occur. In contrast, the critical distance for Gamma-ray bursts is of order 1-kpc. In spite of the high energy effects potentially involved, the geological record provides no clear-cut evidence for any historic supernova induced mass extinctions and/or strong climate change episodes. This, however, is mostly a reflection of their being numerous possible (terrestrial and astronomical) forcing mechanisms acting upon the biosphere and the difficulty of distinguishing between competing scenarios. Key to resolving this situation, it is suggested, is the development of supernova specific extinction and climate change linked ecological models. Moving to the future, we estimate that over the remaining lifetime of the biosphere (˜2 Gyr) the Earth might experience 1 GRB and 20 supernova detonations within their respective harmful threat ranges. There are currently at least 12 potential pre-supernova systems within 1-kpc of the Sun. Of these systems IK Pegasi is the closest Type Ia pre-supernova candidate and Betelgeuse is the closest potential Type II supernova candidate. We review in some detail the past, present and future behavior of these two systems. Developing a detailed evolutionary model we find that IK Pegasi will likely not detonate until some 1.9 billion years hence, and that it affords absolutely no threat to Earth's biosphere. Betelgeuse is the closest, reasonably well understood, pre-supernova candidate to the Sun at the present epoch, and may undergo detonation any time within the next several million years. The stand-off distance of Betelgeuse at the time of its detonation is estimated to fall between 150 and 300-pc—again, affording no possible threat to Earth's biosphere. Temporally, the next most likely, close, potential Type Ic supernova to the Sun is the Wolf-Rayet star within the γ 2 Velorum binary system located at least 260-pc away. It is suggested that evidence relating to large-scale astroengineering projects might fruitfully be looked for in those regions located within 10 to 30-pc of any pre-supernova candidate system.

Implementation of the geoethics principal to environmental technologies by Biogeosystem Technique

NASA Astrophysics Data System (ADS)

Batukaev, Abdulmalik; Kalinitchenko, Valery; Minkina, Tatiana; Mandzhieva, Saglara; Sushkova, Svetlana

2017-04-01

The uncertainty and degradation of biosphere is a result of outdated industrial technologies. The incorrect principals of the nature resources use paradigm are to be radically changed corresponding to principals of Geoethics. Technological dead-end is linked to Philosophy of Technology. The organic protection and imitation of natural patterns are till now the theoretical base of technology. The technological and social determinism are proposed as the "inevitable" for humankind. One is forced to believe that the only way for humanity is to agree that the outdated way of technical development is the only possibility for humankind to survive. But rough imitation as a method of outdated technological platform is fruitless now. Survival under practice of industrial technology platform now has become extremely dangerous. The challenge for humanity is to overcome the chain of environmental hazards of agronomy, irrigation, industry, and other human activities in biosphere, which awkwardly imitate the natural processes: plowing leads to degradation of soil and greenhouse gases emission; irrigation leads to excessive moistening and degradation of soil, landscape, greenhouse gases emission, loss of freshwater - the global deficit; waste utilization leads to greenhouse gases emission, loss of oxigen and other ecological hazards. The fundamentally new technologies are to be generates for development of biosphere, food and resources renewing. Aristotle told that technique can go beyond nature and implement "what nature can't bring to a finish." To overcome fundamental shortcomings of industrial technologies, incorrect land use we propose the Biogeosystem Technique (BGT*) for biosphere sustainability. The BGT* key point is transcendent approach (not imitating of the natural processes) - new technical solutions for biosphere - soil construction, the fluxes of energy, matter, and water control and biological productivity of terrestrial systems. Intra-soil milling which provides the new soil dispersed system synthesis - biological productivity of soil increases twice; intra-soil pulse discrete plants watering which permits to save the freshwater - global deficit - up to 20 times, protect the soil and landscape from excess water, and optimize soil water regime for higher plant's productivity; environmentally safe return of the substances into the active stage of biosphere during synthesis of soil dispersed system and (or) intra-soil pulse discrete plant watering for proper waste recycling. BGT* optimizes an anthropogenic carbon cycle of the Earth, reduces the greenhouse gases emission, implements conditions for green economy, provides an extension of the active area of the biosphere on Earth, water saving, soil and land health. The additional biological product, including food, raw materials and biofuels will be obtained. BGT* can be implemented on the basis of robotics providing cost savings compared to existing industrial technologies of agronomy and environment management. BGT* is the implementation of Geoethics in environmentally safe, productive and low cost technologies of Biosphere at the stage of Noosphere.

Biogeosystem technique as a method to overcome the Biological and Environmental Hazards of modern Agricultural, Irrigational and Technological Activities

NASA Astrophysics Data System (ADS)

Kalinitchenko, Valery; Batukaev, Abdulmalik; Zinchenko, Vladimir; Zarmaev, Ali; Magomadov, Ali; Chernenko, Vladimir; Startsev, Viktor; Bakoev, Serojdin; Dikaev, Zaurbek

2014-05-01

Modern challenge for humanity is to replace the paradigm of nature use and overcome environmental hazards of agronomy, irrigation, industry, and other human activities in biosphere. It is utterly reasonable to stop dividing biosphere on shares - the human habitat and the environment. In the 21st century it is an outdated anthropocentrism. Contradicting himself to biosphere Humankind has the problems. The new paradigm of biosphere control by methods of Biogeosystem technique is on agenda of Humankind. Key directions of Biogeosystem technique. Tillage. Single rotary milling 20…30-50…60 sm soil layer optimizes the evolution and environment of soil, creates a favorable conditions for the rhizosphere, increases the biological productivity of biosphere by 30-50% compared to the standard agricultural practices for the period up to 40 years. Recycle material. Recycling of mineral and organic substances in soil layer of 20…30-50…60 sm in rotary milling soil processing provides wastes clean return to biosphere. Direct intrasoil substances synthesis. Environmentally friendly robot wasteless nanotechnology provides direct substances synthesis, including fertilizers, inside the soil. It eliminates the prerequisites of the wastes formation under standard industrial technologies. Selective substance's extraction from soil. Electrochemical robotic nanotechnology provides selective substances extraction from soil. The technology provides recovery, collection and subsequent safe industrial use of extracted substances out of landscape. Saving fresh water. An important task is to save fresh water in biosphere. Irrigation spends water 4-5 times more of biological requirements of plants, leads to degradation of soil and landscape. The intrasoil pulse continuous-discrete paradigm of irrigation is proposed. It provides the soil and landscape conservation, increases the biological productivity, save the fresh water up to 10-20 times. The subsurface soil rotary processing and intrasoil pulsed continuous-discrete irrigation provide environmentally safe disposal of municipal, industrial, biological and agricultural wastes. Hazardous chemical and biological agents are under the soil surface. It provided a medical and veterinary safety of environment. Biogeosystem technic controls the equilibria in the soil and soil solution, prevents excessive mineralization of organic matter in the surface layers of soil. Simultaneously a soil chemical reduction excluded, biological substance do not degrade to gases. Products of organic matter decomposition are directed to the food chain, 100% waste recycling is obtained. Biogeosystems technique allows producing more biological products hence to recycle excessive amount of man-made CO2 and other substances. Biogeosystems technique increases the rate of photosynthesis of the biosphere, the degree of air ionization. This enhances the formation of rains over land, ensures stability of the ionosphere, magnetosphere and atmosphere of Earth. The nowadays technologies allow applying technical solutions based on Biogeosystem technique, there is unique opportunity to accelerate the noosphere new technological platform.

Weather summaries for Coram Experimental Forest, northwestern Montana-an International Biosphere Reserve

Treesearch

Roger D. Hungerford; Joyce A. Schlieter

1984-01-01

Presents weather data summaries (1934-82) for most of the weather stations within the Coram Experimental Forest (a Biosphere Reserve) in northwestern Montana and for three stations adjacent to the Forest. These data aid in the interpretation of silvicultural and other biological research, particularly the relationships of climatological variations to forest growth and...

Stream denitrification across biomes and its response to anthropogenic nitrate loading

Treesearch

Patrick J Mulholland; Ashely M. Helton; Geoffrey C. Poole; Robert O. Hall; Stephen K. Hamilton; Bruce J. Peterson; Jennifer L. Tank; Linda R. Ashkenas; Lee W. Cooper; Clifford N. Dahm; Walter K. Dodds; Stuart E.G. Findlay; Stanley V. Gregory; Nancy B. Grimm; Sherri L. Johnson; William H. McDowell; Judy L. Meyer; H. Maurice Valett; Jackson R. Webster; Clay P. Arango; Jake J. Beaulieu; Melody J. Bernot; Amy J. Burgin; Chelsea L. Crenshaw; Laura T. Johnson; B.R. Niederlehner; Jonathan M. O' Brien; Jody D. Potter; Richard W. Sheibley; Daniel J. Sobota; Suzanne M. Thomas

2008-01-01

Anthropogenic addition of bioavailable nitrogen to the biosphere is increasing, and terrestrial ecosystems are becoming increasingly nitrogen-saturated, causing more bioavailable nitrogen to enter groundwater and surface waters. Large-scale nitrogen budgets show that an average of about 20 to 25 percent of the nitrogen added to the biosphere is exported from rivers to...

Environmental Art as an Innovative Medium for Environmental Education in Biosphere Reserves

ERIC Educational Resources Information Center

Marks, M.; Chandler, L.; Baldwin, C.

2017-01-01

A key goal of Biosphere Reserves (BR) is to foster environmental education for sustainable development. In this study we systematically analyse two cases in which environmental art is used as a mechanism to engage communities in "building environmental understanding", in Noosa BR in Australia and North Devon BR in the United Kingdom.…

The Loss of Genetic Diversity: An Impending Global Issue.

ERIC Educational Resources Information Center

Pierce, James P.

Definitions of biosphere and ecosystem are provided as the basis for understanding a problem that threatens to become (or already is) a global issue, namely, human activity which results in reducing the diversity of life forms present in the biosphere as an ecosystem. Two aspects of this problem are: (1) the growth of human populations worldwide…

Lunar subsurface architecture enhanced by artificial biosphere concepts

NASA Technical Reports Server (NTRS)

Klassi, Jason D.; Rocha, Carlos J.; Carr, Charles A.

1992-01-01

The integration of artificial biosphere technology with subselene architecture can create a life-enhancing, productive habitat that is safe from solar radiation and extreme temperature fluctuations while maximizing resources brought from Earth and derived from lunar regolith. In the short term, the resulting biotectural (biosphere and architectural) designs will not only make the structures more habitable, productive, and manageable, but will ultimately provide the self-sufficiency factors necessary for the mature lunar settlement. From a long-term perspective, this biotecture approach to astronautics and extraterrestrial development (1) helps reduce mass lift requirements, (2) contributes to habitat self-sufficiency, and (3) actualizes at least one philosophy of solar system exploration, which is to exploit nonterrestrial resources in an effort to conserve our natural resources on this planet.

C-LAMP Subproject Description:Climate Forcing by the Terrestrial Biosphere During the Second Half of the 20th Century

DOE Office of Scientific and Technical Information (OSTI.GOV)

Covey, Curt; Hoffman, Forrest

2008-10-02

This project will quantify selected components of climate forcing due to changes in the terrestrial biosphere over the period 1948-2004, as simulated by the climate / carboncycle models participating in C-LAMP (the Carbon-Land Model Intercomparison Project; see http://www.climatemodeling.org/c-lamp). Unlike other C-LAMP projects that attempt to close the carbon budget, this project will focus on the contributions of individual biomes in terms of the resulting climate forcing. Bala et al. (2007) used a similar (though more comprehensive) model-based technique to assess and compare different components of biospheric climate forcing, but their focus was on potential future deforestation rather than the historicalmore » period.« less

[Is there a connection between biodiversity and the greenhouse effect].

PubMed

Rozanov, S I

1998-01-01

It was discussed the role of biodiversity in ecosystems capacity to control CO2 in atmosphere as the main reason not only of "greenhouse effect" but "greenhouse catastrophe". The necessity to perfect the preventive measures has been defined by time factor. This time may be so little for completing the evolution theory and models of biosphere management. The temps of contemporaneous species extinction exceed two orders as minimum ones how it has been known from planet history. It doesn't permit to discharge that evolutional process will be successful to create organisms which have been capable to stabilize biosphere in conditions of its changing status. It's possible that such change may be provocated with the crisis in civilization-biosphere interrelations.

Egoistic, altruistic, and biospheric environmental concerns: a path analytic investigation of their determinants.

PubMed

Swami, Viren; Chamorro-Premuzic, Tomas; Snelgar, Rosemary; Furnham, Adrian

2010-04-01

Previous studies have shown that environmental concerns (ECs) can be reduced to a three-factor model - comprising altruistic, biospheric, and egoistic concerns - but there have been few studies examining individual difference predictors of ECs. In this study with 203 individuals from a British community sample, we show that biospheric concern was significantly associated with participants' age, political orientation, Machiavellianism, and the Big Five personality traits of Agreeableness, Emotional Stability, and Conscientiousness. Altruistic concern was significantly associated with sex, age, political orientation, and Machiavellianism, but not the Big Five traits, whereas egoistic concern was not significantly associated with any of these predictors except sex. These results are discussed in relation to previous work on ECs and pro-environmental behavior.

Pyrosequencing analysis of the microbial diversity of airag, khoormog and tarag, traditional fermented dairy products of mongolia.

PubMed

Oki, Kaihei; Dugersuren, Jamyan; Demberel, Shirchin; Watanabe, Koichi

2014-01-01

Here, we used pyrosequencing to obtain a detailed analysis of the microbial diversities of traditional fermented dairy products of Mongolia. From 22 Airag (fermented mare's milk), 5 Khoormog (fermented camel's milk) and 26 Tarag (fermented milk of cows, goats and yaks) samples collected in the Mongolian provinces of Arhangai, Bulgan, Dundgobi, Tov, Uburhangai and Umnugobi, we obtained a total of 81 operational taxonomic units, which were assigned to 15 families, 21 genera and 41 species in 3 phyla. The genus Lactobacillus is a core bacterial component of Mongolian fermented milks, and Lactobacillus helveticus, Lactobacillus kefiranofaciens and Lactobacillus delbrueckii were the predominant species of lactic acid bacteria (LAB) in the Airag, Khoormog and Tarag samples, respectively. By using this pyrosequencing approach, we successfully detected most LAB species that have been isolated as well as seven LAB species that have not been found in our previous culture-based study. A subsequent analysis of the principal components of the samples revealed that L. delbrueckii, L. helveticus, L. kefiranofaciens and Streptococcus thermophilus were the main factors influencing the microbial diversity of these Mongolian traditional fermented dairy products and that this diversity correlated with the animal species from which the milk was sourced.

Droplet-based pyrosequencing using digital microfluidics.

PubMed

Boles, Deborah J; Benton, Jonathan L; Siew, Germaine J; Levy, Miriam H; Thwar, Prasanna K; Sandahl, Melissa A; Rouse, Jeremy L; Perkins, Lisa C; Sudarsan, Arjun P; Jalili, Roxana; Pamula, Vamsee K; Srinivasan, Vijay; Fair, Richard B; Griffin, Peter B; Eckhardt, Allen E; Pollack, Michael G

2011-11-15

The feasibility of implementing pyrosequencing chemistry within droplets using electrowetting-based digital microfluidics is reported. An array of electrodes patterned on a printed-circuit board was used to control the formation, transportation, merging, mixing, and splitting of submicroliter-sized droplets contained within an oil-filled chamber. A three-enzyme pyrosequencing protocol was implemented in which individual droplets contained enzymes, deoxyribonucleotide triphosphates (dNTPs), and DNA templates. The DNA templates were anchored to magnetic beads which enabled them to be thoroughly washed between nucleotide additions. Reagents and protocols were optimized to maximize signal over background, linearity of response, cycle efficiency, and wash efficiency. As an initial demonstration of feasibility, a portion of a 229 bp Candida parapsilosis template was sequenced using both a de novo protocol and a resequencing protocol. The resequencing protocol generated over 60 bp of sequence with 100% sequence accuracy based on raw pyrogram levels. Excellent linearity was observed for all of the homopolymers (two, three, or four nucleotides) contained in the C. parapsilosis sequence. With improvements in microfluidic design it is expected that longer reads, higher throughput, and improved process integration (i.e., "sample-to-sequence" capability) could eventually be achieved using this low-cost platform.

Characterization of Bacterial Communities in Venous Insufficiency Wounds by Use of Conventional Culture and Molecular Diagnostic Methods▿

PubMed Central

Tuttle, Marie S.; Mostow, Eliot; Mukherjee, Pranab; Hu, Fen Z.; Melton-Kreft, Rachael; Ehrlich, Garth D.; Dowd, Scot E.; Ghannoum, Mahmoud A.

2011-01-01

Microbial infections delay wound healing, but the effect of the composition of the wound microbiome on healing parameters is unknown. To better understand bacterial communities in chronic wounds, we analyzed debridement samples from lower-extremity venous insufficiency ulcers using the following: conventional anaerobic and aerobic bacterial cultures; the Ibis T5000 universal biosensor (Abbott Molecular); and 16S 454 FLX titanium series pyrosequencing (Roche). Wound debridement samples were obtained from 10 patients monitored clinically for at least 6 months, at which point 5 of the 10 sampled wounds had healed. Pyrosequencing data revealed significantly higher bacterial abundance and diversity in wounds that had not healed at 6 months. Additionally, Actinomycetales was increased in wounds that had not healed, and Pseudomonadaceae was increased in wounds that had healed by the 6-month follow-up. Baseline wound surface area, duration, or analysis by Ibis or conventional culture did not reveal significant differences between wounds that healed after 6 months and those that did not. Thus, pyrosequencing identified distinctive baseline characteristics of wounds that did not heal by the 6-month follow-up, furthering our understanding of potentially unique microbiome characteristics of chronic wounds. PMID:21880958

Massively parallel rRNA gene sequencing exacerbates the potential for biased community diversity comparisons due to variable library sizes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gihring, Thomas; Green, Stefan; Schadt, Christopher Warren

2011-01-01

Technologies for massively parallel sequencing are revolutionizing microbial ecology and are vastly increasing the scale of ribosomal RNA (rRNA) gene studies. Although pyrosequencing has increased the breadth and depth of possible rRNA gene sampling, one drawback is that the number of reads obtained per sample is difficult to control. Pyrosequencing libraries typically vary widely in the number of sequences per sample, even within individual studies, and there is a need to revisit the behaviour of richness estimators and diversity indices with variable gene sequence library sizes. Multiple reports and review papers have demonstrated the bias in non-parametric richness estimators (e.g.more » Chao1 and ACE) and diversity indices when using clone libraries. However, we found that biased community comparisons are accumulating in the literature. Here we demonstrate the effects of sample size on Chao1, ACE, CatchAll, Shannon, Chao-Shen and Simpson's estimations specifically using pyrosequencing libraries. The need to equalize the number of reads being compared across libraries is reiterated, and investigators are directed towards available tools for making unbiased diversity comparisons.« less

The Identification and Differentiation between Burkholderia mallei and Burkholderia pseudomallei Using One Gene Pyrosequencing.

PubMed

Gilling, Damian H; Luna, Vicki Ann; Pflugradt, Cori

2014-01-01

The etiologic agents for melioidosis and glanders, Burkholderia mallei and Burkholderia pseudomallei respectively, are genetically similar making identification and differentiation from other Burkholderia species and each other challenging. We used pyrosequencing to determine the presence or absence of an insertion sequence IS407A within the flagellin P (fliP) gene and to exploit the difference in orientation of this gene in the two species. Oligonucleotide primers were designed to selectively target the IS407A-fliP interface in B. mallei and the fliP gene specifically at the insertion point in B. pseudomallei. We then examined DNA from ten B. mallei, ten B. pseudomallei, 14 B. cepacia, eight other Burkholderia spp., and 17 other bacteria. Resultant pyrograms encompassed the target sequence that contained either the fliP gene with the IS407A interruption or the fully intact fliP gene with 100% sensitivity and 100% specificity. These pyrosequencing assays based upon a single gene enable investigators to reliably identify the two species. The information obtained by these assays provides more knowledge of the genomic reduction that created the new species B. mallei from B. pseudomallei and may point to new targets that can be exploited in the future.

Pyrosequencing-based assessment of microbial community shifts in leachate from animal carcass burial lysimeter.

PubMed

Kim, Hyun Young; Seo, Jiyoung; Kim, Tae-Hun; Shim, Bomi; Cha, Seok Mun; Yu, Seungho

2017-06-01

This study examined the use of microbial community structure as a bio-indicator of decomposition levels. High-throughput pyrosequencing technology was used to assess the shift in microbial community of leachate from animal carcass lysimeter. The leachate samples were collected monthly for one year and a total of 164,639 pyrosequencing reads were obtained and used in the taxonomic classification and operational taxonomy units (OTUs) distribution analysis based on sequence similarity. Our results show considerable changes in the phylum-level bacterial composition, suggesting that the microbial community is a sensitive parameter affected by the burial environment. The phylum classification results showed that Proteobacteria (Pseudomonas) were the most influential taxa in earlier decomposition stage whereas Firmicutes (Clostridium, Sporanaerobacter, and Peptostreptococcus) were dominant in later stage under anaerobic conditions. The result of this study can provide useful information on a time series of leachate profiles of microbial community structures and suggest patterns of microbial diversity in livestock burial sites. In addition, this result can be applicable to predict the decomposition stages under clay loam based soil conditions of animal livestock. Copyright © 2017 Elsevier B.V. All rights reserved.

Droplet-Based Pyrosequencing Using Digital Microfluidics

PubMed Central

Boles, Deborah J.; Benton, Jonathan L.; Siew, Germaine J.; Levy, Miriam H.; Thwar, Prasanna K.; Sandahl, Melissa A.; Rouse, Jeremy L.; Perkins, Lisa C.; Sudarsan, Arjun P.; Jalili, Roxana; Pamula, Vamsee K.; Srinivasan, Vijay; Fair, Richard B.; Griffin, Peter B.; Eckhardt, Allen E.; Pollack, Michael G.

2013-01-01

The feasibility of implementing pyrosequencing chemistry within droplets using electrowetting-based digital microfluidics is reported. An array of electrodes patterned on a printed-circuit board was used to control the formation, transportation, merging, mixing, and splitting of submicroliter-sized droplets contained within an oil-filled chamber. A three-enzyme pyrosequencing protocol was implemented in which individual droplets contained enzymes, deoxyribonucleotide triphosphates (dNTPs), and DNA templates. The DNA templates were anchored to magnetic beads which enabled them to be thoroughly washed between nucleotide additions. Reagents and protocols were optimized to maximize signal over background, linearity of response, cycle efficiency, and wash efficiency. As an initial demonstration of feasibility, a portion of a 229 bp Candida parapsilosis template was sequenced using both a de novo protocol and a resequencing protocol. The resequencing protocol generated over 60 bp of sequence with 100% sequence accuracy based on raw pyrogram levels. Excellent linearity was observed for all of the homopolymers (two, three, or four nucleotides) contained in the C. parapsilosis sequence. With improvements in microfluidic design it is expected that longer reads, higher throughput, and improved process integration (i.e., “sample-to-sequence” capability) could eventually be achieved using this low-cost platform. PMID:21932784

Sensitivity properties of a biosphere model based on BATS and a statistical-dynamical climate model

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, T.

A biosphere model based on the Biosphere-Atmosphere Transfer Scheme (BATS) and the Saltzman-Vernekar (SV) statistical-dynamical climate model is developed. Some equations of BATS are adopted either intact or with modifications, some are conceptually modified, and still others are replaced with equations of the SV model. The model is designed so that it can be run independently as long as the parameters related to the physiology and physiognomy of the vegetation, the atmospheric conditions, solar radiation, and soil conditions are given. With this stand-alone biosphere model, a series of sensitivity investigations, particularly the model sensitivity to fractional area of vegetation cover,more » soil surface water availability, and solar radiation for different types of vegetation, were conducted as a first step. These numerical experiments indicate that the presence of a vegetation cover greatly enhances the exchanges of momentum, water vapor, and energy between the atmosphere and the surface of the earth. An interesting result is that a dense and thick vegetation cover tends to serve as an environment conditioner or, more specifically, a thermostat and a humidistat, since the soil surface temperature, foliage temperature, and temperature and vapor pressure of air within the foliage are practically insensitive to variation of soil surface water availability and even solar radiation within a wide range. An attempt is also made to simulate the gradual deterioration of environment accompanying gradual degradation of a tropical forest to grasslands. Comparison with field data shows that this model can realistically simulate the land surface processes involving biospheric variations. 46 refs., 10 figs., 6 tabs.« less

Sensitivity properties of a biosphere model based on BATS and a statistical-dynamical climate model

NASA Technical Reports Server (NTRS)

Zhang, Taiping

1994-01-01

A biosphere model based on the Biosphere-Atmosphere Transfer Scheme (BATS) and the Saltzman-Vernekar (SV) statistical-dynamical climate model is developed. Some equations of BATS are adopted either intact or with modifications, some are conceptually modified, and still others are replaced with equations of the SV model. The model is designed so that it can be run independently as long as the parameters related to the physiology and physiognomy of the vegetation, the atmospheric conditions, solar radiation, and soil conditions are given. With this stand-alone biosphere model, a series of sensitivity investigations, particularly the model sensitivity to fractional area of vegetation cover, soil surface water availability, and solar radiation for different types of vegetation, were conducted as a first step. These numerical experiments indicate that the presence of a vegetation cover greatly enhances the exchanges of momentum, water vapor, and energy between the atmosphere and the surface of the earth. An interesting result is that a dense and thick vegetation cover tends to serve as an environment conditioner or, more specifically, a thermostat and a humidistat, since the soil surface temperature, foliage temperature, and temperature and vapor pressure of air within the foliage are practically insensitive to variation of soil surface water availability and even solar radiation within a wide range. An attempt is also made to simulate the gradual deterioration of environment accompanying gradual degradation of a tropical forest to grasslands. Comparison with field data shows that this model can realistically simulate the land surface processes involving biospheric variations.

Publications of the biospheric research program: 1981-1987

NASA Technical Reports Server (NTRS)

Wallace, Janice S. (Editor)

1988-01-01

Presented is a list of publications of investigators supported by the Biospheric Research Program of the Biological Systems Research Branch, Life Sciences Division, and the Office of Space Science and Applications. It includes publications dated as of December 31, 1987 and entered into the Life Sciences Bibliographic Database at the George Washington University. Publications are organized by the year published.

"Biosphere Reserve"--The Actual Research Subject of the Sustainable Development Process"

ERIC Educational Resources Information Center

Khasaev, Gabibulla R.; Sadovenko, Marina Yu.; Isaev, Roman O.

2016-01-01

The relevance of the analyzed issue is caused by the growing slippage of research funds of sustainable development in its practice. The purpose of the article is the theoretical basis of the biosphere reserve as a scientific research subject that is relevant to rules of the scientific activity. The leading approach to the study of this issue is…

Bird checklist, Guánica Biosphere Reserve, Puerto Rico

Treesearch

Wayne J. Arendt; John Faaborg; Miguel Canals; Jerry Bauer

2015-01-01

This research note compiles 43 years of research and monitoring data to produce the first comprehensive checklist of the dry forest avian community found within the Guánica Biosphere Reserve. We provide an overview of the reserve along with sighting locales, a list of 185 birds with their resident status and abundance, and a list of the available bird habitats....

Monitoring recreational impacts in wilderness of Kamchatka (on example of Kronotsky State Natural Biosphere Preserve)

Treesearch

Anya V. Zavadskaya

2011-01-01

This paper describes an assessment and monitoring program that was designed and initiated for monitoring recreational impacts in a wilderness in Kamchatka. The framework of the recreational assessment was tested through its application to a case study conducted during the summers of 2008 and 2009 in the Kronotsky State Natural Biosphere Preserve (Kamchatka peninsula,...

Understanding the Relationship between Christian Orthodoxy and Environmentalism : The Mediating Role of Perceived Environmental Consequences

ERIC Educational Resources Information Center

Truelove, Heather Barnes; Joireman, Jeff

2009-01-01

The present study evaluated the hypothesis that people who strongly adhere to Christian orthodoxy may be less proenvironmental to the extent that they are less aware of the biospheric consequences of environmental problems (biospheric AC) but that they may be more proenvironmental than others to the extent that they are more aware of the egoistic…

Astrobiology: Life on Earth (and Elsewhere?)

NASA Technical Reports Server (NTRS)

Des Marais, David J.

2016-01-01

Astrobiology investigates the origins, evolution and distribution of life in the universe. Scientists study how stellar systems and their planets can create planetary environments that sustain biospheres. They search for biosignatures, which are objects, substances and or patterns that indicate the presence of life. Studies of Earth's early biosphere enhance these search strategies and also provide key insights about our own origins.

Searching for Life Underground: An Analysis of Remote Sensing Observations of a Drill Core from Rio Tinto, Spain for Mineralogical Indications of Biological Activity

NASA Technical Reports Server (NTRS)

Battler, M.; Stoker, C.

2005-01-01

Water is unstable on the surface of Mars, and therefore the Martian surface is not likely to support life. It is possible, however, that liquid water exists beneath the surface of Mars, and thus life might also be found in the subsurface. Subsurface life would most likely be microbial, anaerobic, and chemoautotrophic; these types of biospheres on Earth are rare, and not well understood. Finding water and life are high priorities for Mars exploration, and therefore it is important that we learn to explore the subsurface robotically, by drilling. The Mars Analog Rio Tinto Experiment (MARTE), has searched successfully for a subsurface biosphere at Rio Tinto, Spain [1,2,3,4]. The Rio Tinto study site was selected to search for a subsurface biosphere because the extremely low pH and high concentrations of elements such as iron and copper in the Tinto River suggest the presence of a chemoautotrophic biosphere in the subsurface beneath the river. The Rio Tinto has been recognized as an important mineralogical analog to the Sinus Meridiani site on Mars [5].

Methane Production and Transport within the Marsh Biome of Biosphere 2

NASA Technical Reports Server (NTRS)

Molnar, Jennifer; Goodridge, Kelven

1997-01-01

In recent decades, the concentration of methane in the earth's atmosphere increased 1-2% annually. It's rate of increases, combined with methane's effectiveness as a greenhouse gas, has led to an intensive research effort to determine the sources and sinks of the gas in the environment. Biosphere 2 offers a unique opportunity to contribute to the effort because it lacks a major photochemical sink present in the Earth's atmosphere. Researchers can therefore concentrate on biological processes involved in methane cycles. Wetlands are a large source of atmospheric methane, due to anoxic conditions in the sediments and the abundance of organic materials. In order to determine if these conditions in Biosphere 2 also promote methane production, this study looked for the fluxes of methane and methods of transport of the gas from from the water and sediments to the atmosphere in the Marsh Biome. Fluxes of methane from the sediments and waters were measured using static chambers, peepers, and leaf bags. Fluxes and vertical profiles of methane in the sediments show that substantial amounts of methane are being produced in the marsh and are being transported into the Biosphere 2 environment.

The physics, biology, and environmental ethics of making mars habitable.

PubMed

McKay, C P; Marinova, M M

2001-01-01

The considerable evidence that Mars once had a wetter, more clement, environment motivates the search for past or present life on that planet. This evidence also suggests the possibility of restoring habitable conditions on Mars. While the total amounts of the key molecules--carbon dioxide, water, and nitrogen--needed for creating a biosphere on Mars are unknown, estimates suggest that there may be enough in the subsurface. Super greenhouse gases, in particular, perfluorocarbons, are currently the most effective and practical way to warm Mars and thicken its atmosphere so that liquid water is stable on the surface. This process could take approximately 100 years. If enough carbon dioxide is frozen in the South Polar Cap and absorbed in the regolith, the resulting thick and warm carbon dioxide atmosphere could support many types of microorganisms, plants, and invertebrates. If a planet-wide martian biosphere converted carbon dioxide into oxygen with an average efficiency equal to that for Earth's biosphere, it would take > 100,000 years to create Earth-like oxygen levels. Ethical issues associated with bringing life to Mars center on the possibility of indigenous martian life and the relative value of a planet with or without a global biosphere.

Biochemical and physiological bases for the use of carbon and nitrogen isotopes in environmental and ecological studies

NASA Astrophysics Data System (ADS)

Ohkouchi, Naohiko; Ogawa, Nanako O.; Chikaraishi, Yoshito; Tanaka, Hiroyuki; Wada, Eitaro

2015-12-01

We review the biochemical and physiological bases of the use of carbon and nitrogen isotopic compositions as an approach for environmental and ecological studies. Biochemical processes commonly observed in the biosphere, including the decarboxylation and deamination of amino acids, are the key factors in this isotopic approach. The principles drawn from the isotopic distributions disentangle the complex dynamics of the biosphere and allow the interactions between the geosphere and biosphere to be analyzed in detail. We also summarize two recently examined topics with new datasets: the isotopic compositions of individual biosynthetic products (chlorophylls and amino acids) and those of animal organs for further pursuing the basis of the methodology. As a tool for investigating complex systems, compound-specific isotopic analysis compensates the intrinsic disadvantages of bulk isotopic signatures. Chlorophylls provide information about the particular processes of various photoautotrophs, whereas amino acids provide a precise measure of the trophic positions of heterotrophs. The isotopic distributions of carbon and nitrogen in a single organism as well as in the whole biosphere are strongly regulated, so that their major components such as amino acids are coordinated appropriately rather than controlled separately.

The physics, biology, and environmental ethics of making mars habitable

NASA Technical Reports Server (NTRS)

McKay, C. P.; Marinova, M. M.

2001-01-01

The considerable evidence that Mars once had a wetter, more clement, environment motivates the search for past or present life on that planet. This evidence also suggests the possibility of restoring habitable conditions on Mars. While the total amounts of the key molecules--carbon dioxide, water, and nitrogen--needed for creating a biosphere on Mars are unknown, estimates suggest that there may be enough in the subsurface. Super greenhouse gases, in particular, perfluorocarbons, are currently the most effective and practical way to warm Mars and thicken its atmosphere so that liquid water is stable on the surface. This process could take approximately 100 years. If enough carbon dioxide is frozen in the South Polar Cap and absorbed in the regolith, the resulting thick and warm carbon dioxide atmosphere could support many types of microorganisms, plants, and invertebrates. If a planet-wide martian biosphere converted carbon dioxide into oxygen with an average efficiency equal to that for Earth's biosphere, it would take > 100,000 years to create Earth-like oxygen levels. Ethical issues associated with bringing life to Mars center on the possibility of indigenous martian life and the relative value of a planet with or without a global biosphere.

Cytonuclear genetics of experimental fish hybrid zones inside Biosphere 2

USGS Publications Warehouse

Scribner, K.T.; Avise, John C.

1994-01-01

Two species of mosquitofish (family Poeciliidae) known to hybridize in nature were introduced into freshwater habitats inside Biosphere 2, and their population genetics were monitored after 2 years. Within four to six generations, nuclear and cytoplasmic markers characteristic of Gambusia holbrooki had risen greatly in frequency, although some Gambusia affinis alleles and haplotypes were retained primarily in recombinant genotypes, indicative of introgressive hybridization. The temporal cytonuclear dynamics proved similar to population genetic changes observed in replicated experimental hybrid populations outside of Biosphere 2, thus indicating strong directional selection favoring G. holbrooki genotypes across the range of environments monitored. When interpreted in the context of species-specific population demographies observed previously, results suggest that the extremely rapid evolution in these zones of secondary contact is attributable primarily to species differences in life-history traits.

Information in the Biosphere: Biological and Digital Worlds.

PubMed

Gillings, Michael R; Hilbert, Martin; Kemp, Darrell J

2016-03-01

Evolution has transformed life through key innovations in information storage and replication, including RNA, DNA, multicellularity, and culture and language. We argue that the carbon-based biosphere has generated a cognitive system (humans) capable of creating technology that will result in a comparable evolutionary transition. Digital information has reached a similar magnitude to information in the biosphere. It increases exponentially, exhibits high-fidelity replication, evolves through differential fitness, is expressed through artificial intelligence (AI), and has facility for virtually limitless recombination. Like previous evolutionary transitions, the potential symbiosis between biological and digital information will reach a critical point where these codes could compete via natural selection. Alternatively, this fusion could create a higher-level superorganism employing a low-conflict division of labor in performing informational tasks. Copyright © 2015 Elsevier Ltd. All rights reserved.

Data management at Biosphere 2 center

NASA Technical Reports Server (NTRS)

McCreary, Leone F.

1997-01-01

Throughout the history of Biosphere 2, the collecting and recording of biological data has been sporadic. Currently no active effort to administer and record regular biological surveys is being made. Also, there is no central location, such as an on-site data library, where all records from various studies have been archived. As a research institute, good, complete data records are at the core of all Biosphere 2's scientific endeavors. It is therefore imperative that an effective data management system be implemented within the management and research departments as soon as possible. Establishing this system would require three general phases: (1) Design/implement a new archiving/management program (including storage, cataloging and retrieval systems); (2) Organize and input baseline and intermediate data from existing archives; and (3) Maintain records by inputting new data.

Deuterobiosphere the chemosynthetic second biosphere of the globe. A first review.

PubMed

Por, Francis D

2008-06-01

With the discovery of Ophel, a chemosynthesis-based continental subterranean environment (Por 2007), it became evident that such environments, previously known only from the sea, are spanning the world as a parallel biosphere, the deuterobiosphere. In this preliminary contribution the different known biomes of the deuterobiosphere are reviewed and some general and comparative features are listed. © 2008 ISZS, Blackwell Publishing and IOZ/CAS.

Biodiversity conservation, sustainable development, and the U.S. Man and the Biosphere Program: Past contributions and future directions

Treesearch

P. N. Manley; D. C. Hayes

2006-01-01

U.S. Man and the Biosphere (MAB) Program is part of the United Nations Educational, Scientific, and Cultural Organization (UNESCO) MAB program, and is one of six regional MAB programs that span the globe. The MAB Program was created in 1971 with the goal to explore, demonstrate, promote, and encourage harmonious relationships between people and their environments....

Stream denitrification across biomes and its response to anthropogenic nitrate loading

Treesearch

Patrick J. Mulholland; Ashley M. Helton; Geoffrey C. Poole; Robert O. Hall; Stephen K. Hamilton; et al

2008-01-01

Anthropogenic addition of bioavailable nitrogen to the biosphere is increasing1, 2 and terrestrial ecosystems are becoming increasingly nitrogen-saturated3, causing more bioavailable nitrogen to enter groundwater and surface waters4, 5, 6. Large-scale nitrogen budgets show that an average of about 20–25 per cent of the nitrogen added to the biosphere is exported from...

Land-related global habitability science issues

NASA Technical Reports Server (NTRS)

1983-01-01

The scientific investigation of the viewpoint of the biosphere that living organisms and their physical and chemical environment are bound, inseparable parts of one set of closely coupled global processes of the global biogeochemical system, life and life support cycles, is discussed as one of the major scientific challenges of the next decade by building from understanding land processes to interdisciplinary, holistic studies of biospheric dynamics including human impacts.

SeaWiFS Third Anniversary Global Biosphere

NASA Technical Reports Server (NTRS)

2002-01-01

September 18,2000 is the third anniversary of the start of regular SeaWiFS operations of this remarkable planet called Earth. This SeaWiFS image is of the Global Biosphere depicting the ocean's long-term average phytoplankton chlorophyll concentration acquired between September 1997 and August 2000 combined with the SeaWiFS-derived Normalized Difference Vegetation Index (NDVI) over land during July 2000.

Constraining biosphere CO2 flux at regional scale with WRF-CO2 4DVar assimilation system

NASA Astrophysics Data System (ADS)

Zheng, T.

2017-12-01

The WRF-CO2 4DVar assimilation system is updated to include (1) operators for tower based observations (2) chemistry initial and boundary condition in the state vector (3) mechanism for aggregation from simulation model grid to state vector space. The update system is first tested with synthetic data to ensure its accuracy. The system is then used to test regional scale CO2 inversion at MCI (Midcontinental intensive) sites where CO2 mole fraction data were collected at multiple high towers during 2007-2008. The model domain is set to center on Iowa and include 8 towers within its boundary, and it is of 12x12km horizontal grid spacing. First, the relative impacts of the initial and boundary condition are assessed by the system's adjoint model. This is done with 24, 48, 72 hour time span. Second, we assessed the impacts of the transport error, including the misrepresentation of the boundary layer and cumulus activities. Third, we evaluated the different aggregation approach from the native model grid to the control variables (including scaling factors for flux, initial and boundary conditions). Four, we assessed the inversion performance using CO2 observation with different time-interval, and from different tower levels. We also examined the appropriate treatment of the background and observation error covariance in relation with these varying observation data sets.

On the uncertainty of phenological responses to climate change, and implications for a terrestrial biosphere model

NASA Astrophysics Data System (ADS)

Migliavacca, M.; Sonnentag, O.; Keenan, T. F.; Cescatti, A.; O'Keefe, J.; Richardson, A. D.

2012-06-01

Phenology, the timing of recurring life cycle events, controls numerous land surface feedbacks to the climate system through the regulation of exchanges of carbon, water and energy between the biosphere and atmosphere. Terrestrial biosphere models, however, are known to have systematic errors in the simulation of spring phenology, which potentially could propagate to uncertainty in modeled responses to future climate change. Here, we used the Harvard Forest phenology record to investigate and characterize sources of uncertainty in predicting phenology, and the subsequent impacts on model forecasts of carbon and water cycling. Using a model-data fusion approach, we combined information from 20 yr of phenological observations of 11 North American woody species, with 12 leaf bud-burst models that varied in complexity. Akaike's Information Criterion indicated support for spring warming models with photoperiod limitations and, to a lesser extent, models that included chilling requirements. We assessed three different sources of uncertainty in phenological forecasts: parameter uncertainty, model uncertainty, and driver uncertainty. The latter was characterized running the models to 2099 using 2 different IPCC climate scenarios (A1fi vs. B1, i.e. high CO2 emissions vs. low CO2 emissions scenario). Parameter uncertainty was the smallest (average 95% Confidence Interval - CI: 2.4 days century-1 for scenario B1 and 4.5 days century-1 for A1fi), whereas driver uncertainty was the largest (up to 8.4 days century-1 in the simulated trends). The uncertainty related to model structure is also large and the predicted bud-burst trends as well as the shape of the smoothed projections varied among models (±7.7 days century-1 for A1fi, ±3.6 days century-1 for B1). The forecast sensitivity of bud-burst to temperature (i.e. days bud-burst advanced per degree of warming) varied between 2.2 days °C-1 and 5.2 days °C-1 depending on model structure. We quantified the impact of uncertainties in bud-burst forecasts on simulated photosynthetic CO2 uptake and evapotranspiration (ET) using a process-based terrestrial biosphere model. Uncertainty in phenology model structure led to uncertainty in the description of forest seasonality, which accumulated to uncertainty in annual model estimates of gross primary productivity (GPP) and ET of 9.6% and 2.9%, respectively. A sensitivity analysis shows that a variation of ±10 days in bud-burst dates led to a variation of ±5.0% for annual GPP and about ±2.0% for ET. For phenology models, differences among future climate scenarios (i.e. driver) represent the largest source of uncertainty, followed by uncertainties related to model structure, and finally, related to model parameterization. The uncertainties we have quantified will affect the description of the seasonality of ecosystem processes and in particular the simulation of carbon uptake by forest ecosystems, with a larger impact of uncertainties related to phenology model structure, followed by uncertainties related to phenological model parameterization.

Towards Interpreting the Signal of CO2 Emissions from Megacities by Applying a Lagrangian Receptor-oriented Model to OCO-2 XCO2 data

NASA Astrophysics Data System (ADS)

Wu, D.; Lin, J. C.; Oda, T.; Ye, X.; Lauvaux, T.; Yang, E. G.; Kort, E. A.

2017-12-01

Urban regions are large emitters of CO2 whose emission inventories are still associated with large uncertainties. Therefore, a strong need exists to better quantify emissions from megacities using a top-down approach. Satellites — e.g., the Orbiting Carbon Observatory 2 (OCO-2), provide a platform for monitoring spatiotemporal column CO2 concentrations (XCO2). In this study, we present a Lagrangian receptor-oriented model framework and evaluate "model-retrieved" XCO2 by comparing against OCO-2-retrieved XCO2, for three megacities/regions (Riyadh, Cairo and Pearl River Delta). OCO-2 soundings indicate pronounced XCO2 enhancements (dXCO2) when crossing Riyadh, which are successfully captured by our model with a slight latitude shift. From this model framework, we can identify and compare the relative contributions of dXCO2 resulted from anthropogenic emission versus biospheric fluxes. In addition, to impose constraints on emissions for Riyadh through inversion methods, three uncertainties sources are addressed in this study, including 1) transport errors, 2) receptor and model setups in atmospheric models, and 3) urban emission uncertainties. For 1), we calculate transport errors by adding a wind error component to randomize particle distributions. For 2), a set of sensitivity tests using bootstrap method is performed to describe proper ways to setup receptors in Lagrangian models. For 3), both emission uncertainties from the Fossil Fuel Data Assimilation System (FFDAS) and the spread among three emission inventories are used to approximate an overall fractional uncertainty in modeled anthropogenic signal (dXCO2.anthro). Lastly, we investigate the definition of background (clean) XCO2 for megacities from retrieved XCO2 by means of statistical tools and our model framework.

Is there a shadow biosphere of silicon utilizing organisms on earth?

NASA Astrophysics Data System (ADS)

Das, P.; Das, S.

2012-12-01

The idea of shadow biosphere was first visualized by Carol Cleland and Shelley Copley of the University of Colorado in 2005. Since that time many scientists were interested in this subject and the basis of this paper is also correlated with this novel idea. A shadow biosphere essentially consists of self replicating bodies utilizing one or more chemicals which are not exploited to such extent by organisms of the real biosphere. Silicon-utilizing organisms mainly belong to the diatoms, some other algae, sponges, monocotyledon plants in this category are classified by us. They can survive in extremes of temperature, pressure, radiation, pH, salinity and nutrient conditions and this unusual tolerance to stress depends on their silicon biomineralization occurring either as a biologically controlled process or as a biologically induced process. In the control process silicate is sequestered and transferred to the intracellular silica deposition vesicle by energy driven pump mechanism in presence of specific transporter protein. In induced silicon mineralization, initial amorphous nucleation of silicon is gradually transformed to a crystalline phase. Besides these two methods, there are also other processes in which formation of silanol and siloxane can occur from oligomers of silica originating from monomers by Hydrogen bonding, cation bridging and direct electrostatic interactions. The structure of the silica biomineralised cell wall in silicon utilizing organisms is very complex, but this may explain their ability to stress tolerance and their possible coordinating role with other essential bioactive elements in nature. When electropositive elements are less, then polymerization of silicon-oxygen excess may occur easily, particularly in carbon and nitrogen paucity in the environment. Thus all these features indicate possibility of a shadow biosphere of these organisms on earth.

The Potential Feasibility of Chlorinic Photosynthesis on Exoplanets

NASA Astrophysics Data System (ADS)

Haas, Johnson R.

2010-11-01

The modern search for life-bearing exoplanets emphasizes the potential detection of O2 and O3 absorption spectra in exoplanetary atmospheres as ideal signatures of biology. However, oxygenic photosynthesis may not arise ubiquitously in exoplanetary biospheres. Alternative evolutionary paths may yield planetary atmospheres tinted with the waste products of other dominant metabolisms, including potentially exotic biochemistries. This paper defines chlorinic photosynthesis (CPS) as biologically mediated photolytic oxidation of aqueous Cl- to form halocarbon or dihalogen products, coupled with CO2 assimilation. This hypothetical metabolism appears to be feasible energetically, physically, and geochemically, and could potentially develop under conditions that approximate the terrestrial Archean. It is hypothesized that an exoplanetary biosphere in which chlorinic photosynthesis dominates primary production would tend to evolve a strongly oxidizing, halogen-enriched atmosphere over geologic time. It is recommended that astronomical observations of exoplanetary outgoing thermal emission spectra consider signs of halogenated chemical species as likely indicators of the presence of a chlorinic biosphere. Planets that favor the evolution of CPS would probably receive equivalent or greater surface UV flux than is produced by the Sun, which would promote stronger abiotic UV photolysis of aqueous halides than occurred during Earth's Archean era and impose stronger evolutionary selection pressures on endemic life to accommodate and utilize halogenated compounds. Ocean-bearing planets of stars with metallicities equivalent to, or greater than, the Sun should especially favor the evolution of chlorinic biospheres because of the higher relative seawater abundances of Cl, Br, and I such planets would tend to host. Directed searches for chlorinic biospheres should probably focus on G0-G2, F, and A spectral class stars that have bulk metallicities of +0.0 Dex or greater.

Biospheric feedback effects in a synchronously coupled model of human and Earth systems

DOE Office of Scientific and Technical Information (OSTI.GOV)

Thornton, Peter E.; Calvin, Katherine; Jones, Andrew D.

Fossil fuel combustion and land-use change are the first and second largest contributors to industrial-era increases in atmospheric carbon dioxide concentration, which is itself the largest driver of present-day climate change1. Projections of fossil fuel consumption and land-use change are thus fundamental inputs for coupled Earth system models (ESM) used to estimate the physical and biological consequences of future climate system forcing2,3. While empirical datasets are available to inform historical analyses4,5, assessments of future climate change have relied on projections of energy and land use based on energy economic models, constrained using historical and present-day data and forced with assumptionsmore » about future policy, land-use patterns, and socio-economic development trajectories6. Here we show that the influence of biospheric change – the integrated effect of climatic, ecological, and geochemical processes – on land ecosystems has a significant impact on energy, agriculture, and land-use projections for the 21st century. Such feedbacks have been ignored in previous ESM studies of future climate. We find that synchronous exposure of land ecosystem productivity in the economic system to biospheric change as it develops in an ESM results in a 10% reduction of land area used for crop cultivation; increased managed forest area and land carbon; a 15-20% decrease in global crop price; and a 17% reduction in fossil fuel emissions for a low-mid range forcing scenario7. These simulation results demonstrate that biospheric change can significantly alter primary human system forcings to the climate system. This synchronous two-way coupling approach removes inconsistencies in description of climate change between human and biosphere components of the coupled model, mitigating a major source of uncertainty identified in assessments of future climate projections8-10.« less

The potential feasibility of chlorinic photosynthesis on exoplanets.

PubMed

Haas, Johnson R

2010-11-01

The modern search for life-bearing exoplanets emphasizes the potential detection of O(2) and O(3) absorption spectra in exoplanetary atmospheres as ideal signatures of biology. However, oxygenic photosynthesis may not arise ubiquitously in exoplanetary biospheres. Alternative evolutionary paths may yield planetary atmospheres tinted with the waste products of other dominant metabolisms, including potentially exotic biochemistries. This paper defines chlorinic photosynthesis (CPS) as biologically mediated photolytic oxidation of aqueous Cl(-) to form halocarbon or dihalogen products, coupled with CO(2) assimilation. This hypothetical metabolism appears to be feasible energetically, physically, and geochemically, and could potentially develop under conditions that approximate the terrestrial Archean. It is hypothesized that an exoplanetary biosphere in which chlorinic photosynthesis dominates primary production would tend to evolve a strongly oxidizing, halogen-enriched atmosphere over geologic time. It is recommended that astronomical observations of exoplanetary outgoing thermal emission spectra consider signs of halogenated chemical species as likely indicators of the presence of a chlorinic biosphere. Planets that favor the evolution of CPS would probably receive equivalent or greater surface UV flux than is produced by the Sun, which would promote stronger abiotic UV photolysis of aqueous halides than occurred during Earth's Archean era and impose stronger evolutionary selection pressures on endemic life to accommodate and utilize halogenated compounds. Ocean-bearing planets of stars with metallicities equivalent to, or greater than, the Sun should especially favor the evolution of chlorinic biospheres because of the higher relative seawater abundances of Cl, Br, and I such planets would tend to host. Directed searches for chlorinic biospheres should probably focus on G0-G2, F, and A spectral class stars that have bulk metallicities of +0.0 Dex or greater.

Turnover of microbial lipids in the deep biosphere and growth of benthic archaeal populations

PubMed Central

Xie, Sitan; Lipp, Julius S.; Wegener, Gunter; Ferdelman, Timothy G.; Hinrichs, Kai-Uwe

2013-01-01

Deep subseafloor sediments host a microbial biosphere with unknown impact on global biogeochemical cycles. This study tests previous evidence based on microbial intact polar lipids (IPLs) as proxies of live biomass, suggesting that Archaea dominate the marine sedimentary biosphere. We devised a sensitive radiotracer assay to measure the decay rate of ([14C]glucosyl)-diphytanylglyceroldiether (GlcDGD) as an analog of archaeal IPLs in continental margin sediments. The degradation kinetics were incorporated in model simulations that constrained the fossil fraction of subseafloor IPLs and rates of archaeal turnover. Simulating the top 1 km in a generic continental margin sediment column, we estimated degradation rate constants of GlcDGD being one to two orders of magnitude lower than those of bacterial IPLs, with half-lives of GlcDGD increasing with depth to 310 ky. Given estimated microbial community turnover times of 1.6–73 ky in sediments deeper than 1 m, 50–96% of archaeal IPLs represent fossil signals. Consequently, previous lipid-based estimates of global subseafloor biomass probably are too high, and the widely observed dominance of archaeal IPLs does not rule out a deep biosphere dominated by Bacteria. Reverse modeling of existing concentration profiles suggest that archaeal IPL synthesis rates decline from around 1,000 pg⋅mL−1 sediment⋅y−1 at the surface to 0.2 pg⋅mL−1⋅y−1 at 1 km depth, equivalent to production of 7 × 105 to 140 archaeal cells⋅mL−1 sediment⋅y−1, respectively. These constraints on microbial growth are an important step toward understanding the relationship between the deep biosphere and the carbon cycle. PMID:23530229

Earth Applications of Closed Ecological Systems: Relevance to the Development of Sustainability in our Global Biosphere

NASA Astrophysics Data System (ADS)

Dempster, W.; van Thillo, M.; Alling, A.; Allen, J.; Silverstone, S.; Nelson, M.

The parallels between the challenges facing bioregenerative life support and closed ecological systems and those in our global biosphere are striking. At the scale of the current global technosphere and human population, it is increasingly obvious that the biosphere can no longer be counted on to be vast enough to safely buffer and absorb technogenic and anthropogenic pollutants. With an increasing percentage of the world's natural resources and primary productivity being dictated by, and directed to, humans, our species is starting to appreciate its survival and quality of life depends on regulating its activities, and insuring that crucial biogeochemical cycles continue to function. This shift of consciousness has led to the widespread call for moving towards the sustainability of human activities. For researchers working on bioreenerative life support, the small volumes and faster cycling times have made it obvious that systems must be created in to ensure renewal of water and atmosphere, nutrient recycling, and where all technical systems can be safely integrated with the maintenance of safe environmental conditions. The development of technical systems that can be fully integrated with the living systems that they support should be a harbinger of new perspectives in the global environment. The paper will review some of these environmental technologies which are emerging from bioregenerative life support system research such as high-yield intensive agricultural methods, waste treatment and nutrient recycling, air purification, modeling, sensor and control systems and their potential applications in the global biosphere. In addition, a review of the human experience in closed ecological systems shows that these can offer opportunities for public education and consciousness-changing of how humans regard our global biosphere.

Predicting ecosystem dynamics at regional scales: an evaluation of a terrestrial biosphere model for the forests of northeastern North America.

PubMed

Medvigy, David; Moorcroft, Paul R

2012-01-19

Terrestrial biosphere models are important tools for diagnosing both the current state of the terrestrial carbon cycle and forecasting terrestrial ecosystem responses to global change. While there are a number of ongoing assessments of the short-term predictive capabilities of terrestrial biosphere models using flux-tower measurements, to date there have been relatively few assessments of their ability to predict longer term, decadal-scale biomass dynamics. Here, we present the results of a regional-scale evaluation of the Ecosystem Demography version 2 (ED2)-structured terrestrial biosphere model, evaluating the model's predictions against forest inventory measurements for the northeast USA and Quebec from 1985 to 1995. Simulations were conducted using a default parametrization, which used parameter values from the literature, and a constrained model parametrization, which had been developed by constraining the model's predictions against 2 years of measurements from a single site, Harvard Forest (42.5° N, 72.1° W). The analysis shows that the constrained model parametrization offered marked improvements over the default model formulation, capturing large-scale variation in patterns of biomass dynamics despite marked differences in climate forcing, land-use history and species-composition across the region. These results imply that data-constrained parametrizations of structured biosphere models such as ED2 can be successfully used for regional-scale ecosystem prediction and forecasting. We also assess the model's ability to capture sub-grid scale heterogeneity in the dynamics of biomass growth and mortality of different sizes and types of trees, and then discuss the implications of these analyses for further reducing the remaining biases in the model's predictions.

Turnover of microbial lipids in the deep biosphere and growth of benthic archaeal populations.

PubMed

Xie, Sitan; Lipp, Julius S; Wegener, Gunter; Ferdelman, Timothy G; Hinrichs, Kai-Uwe

2013-04-09

Deep subseafloor sediments host a microbial biosphere with unknown impact on global biogeochemical cycles. This study tests previous evidence based on microbial intact polar lipids (IPLs) as proxies of live biomass, suggesting that Archaea dominate the marine sedimentary biosphere. We devised a sensitive radiotracer assay to measure the decay rate of ([(14)C]glucosyl)-diphytanylglyceroldiether (GlcDGD) as an analog of archaeal IPLs in continental margin sediments. The degradation kinetics were incorporated in model simulations that constrained the fossil fraction of subseafloor IPLs and rates of archaeal turnover. Simulating the top 1 km in a generic continental margin sediment column, we estimated degradation rate constants of GlcDGD being one to two orders of magnitude lower than those of bacterial IPLs, with half-lives of GlcDGD increasing with depth to 310 ky. Given estimated microbial community turnover times of 1.6-73 ky in sediments deeper than 1 m, 50-96% of archaeal IPLs represent fossil signals. Consequently, previous lipid-based estimates of global subseafloor biomass probably are too high, and the widely observed dominance of archaeal IPLs does not rule out a deep biosphere dominated by Bacteria. Reverse modeling of existing concentration profiles suggest that archaeal IPL synthesis rates decline from around 1,000 pg⋅mL(-1) sediment⋅y(-1) at the surface to 0.2 pg⋅mL(-1)⋅y(-1) at 1 km depth, equivalent to production of 7 × 10(5) to 140 archaeal cells⋅mL(-1) sediment⋅y(-1), respectively. These constraints on microbial growth are an important step toward understanding the relationship between the deep biosphere and the carbon cycle.

Atribacteria from the Subseafloor Sedimentary Biosphere Disperse to the Hydrosphere through Submarine Mud Volcanoes.

PubMed

Hoshino, Tatsuhiko; Toki, Tomohiro; Ijiri, Akira; Morono, Yuki; Machiyama, Hideaki; Ashi, Juichiro; Okamura, Kei; Inagaki, Fumio

2017-01-01

Submarine mud volcanoes (SMVs) are formed by muddy sediments and breccias extruded to the seafloor from a source in the deep subseafloor and are characterized by the discharge of methane and other hydrocarbon gasses and deep-sourced fluids into the overlying seawater. Although SMVs act as a natural pipeline connecting the Earth's surface and subsurface biospheres, the dispersal of deep-biosphere microorganisms and their ecological roles remain largely unknown. In this study, we investigated the microbial communities in sediment and overlying seawater at two SMVs located on the Ryukyu Trench off Tanegashima Island, southern Japan. The microbial communities in mud volcano sediments were generally distinct from those in the overlying seawaters and in the well-stratified Pacific margin sediments collected at the Peru Margin, the Juan de Fuca Ridge flank off Oregon, and offshore of Shimokita Peninsula, northeastern Japan. Nevertheless, in-depth analysis of different taxonomic groups at the sub-species level revealed that the taxon affiliated with Atribacteria , heterotrophic anaerobic bacteria that typically occur in organic-rich anoxic subseafloor sediments, were commonly found not only in SMV sediments but also in the overlying seawater. We designed a new oligonucleotide probe for detecting Atribacteria using the catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH). CARD-FISH, digital PCR and sequencing analysis of 16S rRNA genes consistently showed that Atribacteria are abundant in the methane plumes of the two SMVs (0.58 and 1.5 × 10 4 cells/mL, respectively) but not in surrounding waters, suggesting that microbial cells in subseafloor sediments are dispersed as "deep-biosphere seeds" into the ocean. These findings may have important implications for the microbial transmigration between the deep subseafloor biosphere and the hydrosphere.

Atribacteria from the Subseafloor Sedimentary Biosphere Disperse to the Hydrosphere through Submarine Mud Volcanoes

PubMed Central

Hoshino, Tatsuhiko; Toki, Tomohiro; Ijiri, Akira; Morono, Yuki; Machiyama, Hideaki; Ashi, Juichiro; Okamura, Kei; Inagaki, Fumio

2017-01-01

Submarine mud volcanoes (SMVs) are formed by muddy sediments and breccias extruded to the seafloor from a source in the deep subseafloor and are characterized by the discharge of methane and other hydrocarbon gasses and deep-sourced fluids into the overlying seawater. Although SMVs act as a natural pipeline connecting the Earth’s surface and subsurface biospheres, the dispersal of deep-biosphere microorganisms and their ecological roles remain largely unknown. In this study, we investigated the microbial communities in sediment and overlying seawater at two SMVs located on the Ryukyu Trench off Tanegashima Island, southern Japan. The microbial communities in mud volcano sediments were generally distinct from those in the overlying seawaters and in the well-stratified Pacific margin sediments collected at the Peru Margin, the Juan de Fuca Ridge flank off Oregon, and offshore of Shimokita Peninsula, northeastern Japan. Nevertheless, in-depth analysis of different taxonomic groups at the sub-species level revealed that the taxon affiliated with Atribacteria, heterotrophic anaerobic bacteria that typically occur in organic-rich anoxic subseafloor sediments, were commonly found not only in SMV sediments but also in the overlying seawater. We designed a new oligonucleotide probe for detecting Atribacteria using the catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH). CARD-FISH, digital PCR and sequencing analysis of 16S rRNA genes consistently showed that Atribacteria are abundant in the methane plumes of the two SMVs (0.58 and 1.5 × 104 cells/mL, respectively) but not in surrounding waters, suggesting that microbial cells in subseafloor sediments are dispersed as “deep-biosphere seeds” into the ocean. These findings may have important implications for the microbial transmigration between the deep subseafloor biosphere and the hydrosphere. PMID:28676800

Estimation of Pre-industrial Nitrous Oxide Emission from the Terrestrial Biosphere

NASA Astrophysics Data System (ADS)

Xu, R.; Tian, H.; Lu, C.; Zhang, B.; Pan, S.; Yang, J.

2015-12-01

Nitrous oxide (N2O) is currently the third most important greenhouse gases (GHG) after methane (CH4) and carbon dioxide (CO2). Global N2O emission increased substantially primarily due to reactive nitrogen (N) enrichment through fossil fuel combustion, fertilizer production, and legume crop cultivation etc. In order to understand how climate system is perturbed by anthropogenic N2O emissions from the terrestrial biosphere, it is necessary to better estimate the pre-industrial N2O emissions. Previous estimations of natural N2O emissions from the terrestrial biosphere range from 3.3-9.0 Tg N2O-N yr-1. This large uncertainty in the estimation of pre-industrial N2O emissions from the terrestrial biosphere may be caused by uncertainty associated with key parameters such as maximum nitrification and denitrification rates, half-saturation coefficients of soil ammonium and nitrate, N fixation rate, and maximum N uptake rate. In addition to the large estimation range, previous studies did not provide an estimate on preindustrial N2O emissions at regional and biome levels. In this study, we applied a process-based coupled biogeochemical model to estimate the magnitude and spatial patterns of pre-industrial N2O fluxes at biome and continental scales as driven by multiple input data, including pre-industrial climate data, atmospheric CO2 concentration, N deposition, N fixation, and land cover types and distributions. Uncertainty associated with key parameters is also evaluated. Finally, we generate sector-based estimates of pre-industrial N2O emission, which provides a reference for assessing the climate forcing of anthropogenic N2O emission from the land biosphere.

Pathogenic bacteria in sewage treatment plants as revealed by 454 pyrosequencing.

PubMed

Ye, Lin; Zhang, Tong

2011-09-01

This study applied 454 high-throughput pyrosequencing to analyze potentially pathogenic bacteria in activated sludge from 14 municipal wastewater treatment plants (WWTPs) across four countries (China, U.S., Canada, and Singapore), plus the influent and effluent of one of the 14 WWTPs. A total of 370,870 16S rRNA gene sequences with average length of 207 bps were obtained and all of them were assigned to corresponding taxonomic ranks by using RDP classifier and MEGAN. It was found that the most abundant potentially pathogenic bacteria in the WWTPs were affiliated with the genera of Aeromonas and Clostridium. Aeromonas veronii, Aeromonas hydrophila, and Clostridium perfringens were species most similar to the potentially pathogenic bacteria found in this study. Some sequences highly similar (>99%) to Corynebacterium diphtheriae were found in the influent and activated sludge samples from a saline WWTP. Overall, the percentage of the sequences closely related (>99%) to known pathogenic bacteria sequences was about 0.16% of the total sequences. Additionally, a platform-independent Java application (BAND) was developed for graphical visualization of the data of microbial abundance generated by high-throughput pyrosequencing. The approach demonstrated in this study could examine most of the potentially pathogenic bacteria simultaneously instead of one-by-one detection by other methods.

Pyrosequencing Analysis of the Microbial Diversity of Airag, Khoormog and Tarag, Traditional Fermented Dairy Products of Mongolia

PubMed Central

OKI, Kaihei; DUGERSUREN, Jamyan; DEMBEREL, Shirchin; WATANABE, Koichi

2014-01-01

Here, we used pyrosequencing to obtain a detailed analysis of the microbial diversities of traditional fermented dairy products of Mongolia. From 22 Airag (fermented mare’s milk), 5 Khoormog (fermented camel’s milk) and 26 Tarag (fermented milk of cows, goats and yaks) samples collected in the Mongolian provinces of Arhangai, Bulgan, Dundgobi, Tov, Uburhangai and Umnugobi, we obtained a total of 81 operational taxonomic units, which were assigned to 15 families, 21 genera and 41 species in 3 phyla. The genus Lactobacillus is a core bacterial component of Mongolian fermented milks, and Lactobacillus helveticus, Lactobacillus kefiranofaciens and Lactobacillus delbrueckii were the predominant species of lactic acid bacteria (LAB) in the Airag, Khoormog and Tarag samples, respectively. By using this pyrosequencing approach, we successfully detected most LAB species that have been isolated as well as seven LAB species that have not been found in our previous culture-based study. A subsequent analysis of the principal components of the samples revealed that L. delbrueckii, L. helveticus, L. kefiranofaciens and Streptococcus thermophilus were the main factors influencing the microbial diversity of these Mongolian traditional fermented dairy products and that this diversity correlated with the animal species from which the milk was sourced. PMID:25003019

Pyrosequencing the Manduca sexta larval midgut transcriptome: messages for digestion, detoxification and defence.

PubMed

Pauchet, Y; Wilkinson, P; Vogel, H; Nelson, D R; Reynolds, S E; Heckel, D G; ffrench-Constant, R H

2010-02-01

The tobacco hornworm Manduca sexta is an important model for insect physiology but genomic and transcriptomic data are currently lacking. Following a recent pyrosequencing study generating immune related expressed sequence tags (ESTs), here we use this new technology to define the M. sexta larval midgut transcriptome. We generated over 387,000 midgut ESTs, using a combination of Sanger and 454 sequencing, and classified predicted proteins into those involved in digestion, detoxification and immunity. In many cases the depth of 454 pyrosequencing coverage allowed us to define the entire cDNA sequence of a particular gene. Many new M. sexta genes are described including up to 36 new cytochrome P450s, some of which have been implicated in the metabolism of host plant-derived nicotine. New lepidopteran gene families such as the beta-fructofuranosidases, previously thought to be restricted to Bombyx mori, are also described. An unexpectedly high number of ESTs were involved in immunity, for example 39 contigs encoding serpins, and the increasingly appreciated role of the midgut in insect immunity is discussed. Similar studies of other tissues will allow for a tissue by tissue description of the M. sexta transcriptome and will form an essential complimentary step on the road to genome sequencing and annotation.

DNA Methylation Pyrosequencing Assay Is Applicable for the Assessment of Epigenetic Active Environmental or Clinical Relevant Chemicals

PubMed Central

Florea, Ana-Maria

2013-01-01

Exposure of cells and organisms to stressors might result in epigenetic changes. Here it is shown that investigation of DNA methylation using pyrosequencing is an alternative for in vitro and in vivo toxicological testing of epigenetic effects induced by chemicals and drugs. An in vitro evaluation of global and CpG site specific DNA methylation upon treatment of cells with chemicals/drugs is shown. Bisulfite genomic sequencing of methylation controls showed high methylation of LINE1 in methylation positive control and low methylation in the negative controls. The CpG sites within the LINE1 element are methylated at different levels. In vitro cell cultures show a methylation level ranging from 56% to 49%. Cultures of drug resistant tumor cells show significant hypomethylation as compared with the originating nonresistant tumor cells. The in vitro testing of epigenetically active chemicals (5-methyl-2'-deoxycytidine and trichostatin A) revealed a significant change of LINE1 methylation status upon treatment, while specific CpG sites were more prone to demethylation than others (focal methylation). In conclusion, DNA methylation using pyrosequencing might be used not only for testing epigenetic toxins/drugs but also in risk assessment of drugs, food, and environmental relevant pollutants. PMID:24093099

[Effect of X-ray micro-computed tomography on the metabolic activity and diversity of soil microbial communities in two Chinese soils].

PubMed

Zu, Qianhui; Fang, Huan; Zhou, Hu; Zhang, Jianwei; Peng, Xinhua; Lin, Xiangui; Feng, Youzhi

2016-01-04

X-ray micro-computed tomography (micro-CT) technology, as used in the in situ and nondestructive analysis of soil physical structure, provides the opportunity of associating soil physical and biological assays. Due to the high heterogeneity of the soil matrix, X-ray micro-CT scanning and soil microbial assays should be conducted on the same soil sample. This raises the question whether X-ray micro-CT influences microbial function and diversity of the sample soil to be analyzed. To address this question, we used plate counting, microcalorimetry and pyrosequencing approaches to evaluate the effect of X-ray--at doses typically used in micro-CT--on soil microorganisms in a typical soil of North China Plain, Fluvo-aquic soil and in a typical soil of subtropical China, Ultisol soil, respectively. In both soils radiation decreased the number of viable soil bacteria and disturbed their thermogenic profiles. At DNA level, pyrosequencing revealed that alpha diversities of two soils biota were influenced in opposite ways, while beta diversity was not affected although the relative abundances of some guilds were changed. These findings indicate that the metabolically active aspects of soil biota are not compatible with X-ray micro-CT; while the beta molecular diversity based on pyrosequencing could be compatible.

454 pyrosequencing analyses of bacterial and archaeal richness in 21 full-scale biogas digesters.

PubMed

Sundberg, Carina; Al-Soud, Waleed A; Larsson, Madeleine; Alm, Erik; Yekta, Sepehr S; Svensson, Bo H; Sørensen, Søren J; Karlsson, Anna

2013-09-01

The microbial community of 21 full-scale biogas reactors was examined using 454 pyrosequencing of 16S rRNA gene sequences. These reactors included seven (six mesophilic and one thermophilic) digesting sewage sludge (SS) and 14 (ten mesophilic and four thermophilic) codigesting (CD) various combinations of wastes from slaughterhouses, restaurants, households, etc. The pyrosequencing generated more than 160,000 sequences representing 11 phyla, 23 classes, and 95 genera of Bacteria and Archaea. The bacterial community was always both more abundant and more diverse than the archaeal community. At the phylum level, the foremost populations in the SS reactors included Actinobacteria, Proteobacteria, Chloroflexi, Spirochetes, and Euryarchaeota, while Firmicutes was the most prevalent in the CD reactors. The main bacterial class in all reactors was Clostridia. Acetoclastic methanogens were detected in the SS, but not in the CD reactors. Their absence suggests that methane formation from acetate takes place mainly via syntrophic acetate oxidation in the CD reactors. A principal component analysis of the communities at genus level revealed three clusters: SS reactors, mesophilic CD reactors (including one thermophilic CD and one SS), and thermophilic CD reactors. Thus, the microbial composition was mainly governed by the substrate differences and the process temperature. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

The Identification and Differentiation between Burkholderia mallei and Burkholderia pseudomallei Using One Gene Pyrosequencing

PubMed Central

Gilling, Damian H.; Luna, Vicki Ann; Pflugradt, Cori

2014-01-01

The etiologic agents for melioidosis and glanders, Burkholderia mallei and Burkholderia pseudomallei respectively, are genetically similar making identification and differentiation from other Burkholderia species and each other challenging. We used pyrosequencing to determine the presence or absence of an insertion sequence IS407A within the flagellin P (fliP) gene and to exploit the difference in orientation of this gene in the two species. Oligonucleotide primers were designed to selectively target the IS407A-fliP interface in B. mallei and the fliP gene specifically at the insertion point in B. pseudomallei. We then examined DNA from ten B. mallei, ten B. pseudomallei, 14 B. cepacia, eight other Burkholderia spp., and 17 other bacteria. Resultant pyrograms encompassed the target sequence that contained either the fliP gene with the IS407A interruption or the fully intact fliP gene with 100% sensitivity and 100% specificity. These pyrosequencing assays based upon a single gene enable investigators to reliably identify the two species. The information obtained by these assays provides more knowledge of the genomic reduction that created the new species B. mallei from B. pseudomallei and may point to new targets that can be exploited in the future. PMID:27350960

Rapid detection and subtyping of human influenza A viruses and reassortants by pyrosequencing.

PubMed

Deng, Yi-Mo; Caldwell, Natalie; Barr, Ian G

2011-01-01

Given the continuing co-circulation of the 2009 H1N1 pandemic influenza A viruses with seasonal H3N2 viruses, rapid and reliable detection of newly emerging influenza reassortant viruses is important to enhance our influenza surveillance. A novel pyrosequencing assay was developed for the rapid identification and subtyping of potential human influenza A virus reassortants based on all eight gene segments of the virus. Except for HA and NA genes, one universal set of primers was used to amplify and subtype each of the six internal genes. With this method, all eight gene segments of 57 laboratory isolates and 17 original specimens of seasonal H1N1, H3N2 and 2009 H1N1 pandemic viruses were correctly matched with their corresponding subtypes. In addition, this method was shown to be capable of detecting reassortant viruses by correctly identifying the source of all 8 gene segments from three vaccine production reassortant viruses and three H1N2 viruses. In summary, this pyrosequencing assay is a sensitive and specific procedure for screening large numbers of viruses for reassortment events amongst the commonly circulating human influenza A viruses, which is more rapid and cheaper than using conventional sequencing approaches.

Rapid Detection and Subtyping of Human Influenza A Viruses and Reassortants by Pyrosequencing

PubMed Central

Deng, Yi-Mo; Caldwell, Natalie; Barr, Ian G.

2011-01-01

Background Given the continuing co-circulation of the 2009 H1N1 pandemic influenza A viruses with seasonal H3N2 viruses, rapid and reliable detection of newly emerging influenza reassortant viruses is important to enhance our influenza surveillance. Methodology/Principal Findings A novel pyrosequencing assay was developed for the rapid identification and subtyping of potential human influenza A virus reassortants based on all eight gene segments of the virus. Except for HA and NA genes, one universal set of primers was used to amplify and subtype each of the six internal genes. With this method, all eight gene segments of 57 laboratory isolates and 17 original specimens of seasonal H1N1, H3N2 and 2009 H1N1 pandemic viruses were correctly matched with their corresponding subtypes. In addition, this method was shown to be capable of detecting reassortant viruses by correctly identifying the source of all 8 gene segments from three vaccine production reassortant viruses and three H1N2 viruses. Conclusions/Significance In summary, this pyrosequencing assay is a sensitive and specific procedure for screening large numbers of viruses for reassortment events amongst the commonly circulating human influenza A viruses, which is more rapid and cheaper than using conventional sequencing approaches. PMID:21886790

Analysis of Gastric Microbiota by Pyrosequencing: Minor Role of Bacteria Other Than Helicobacter pylori in the Gastric Carcinogenesis.

PubMed

Jo, Hyun Jin; Kim, Jaeyeon; Kim, Nayoung; Park, Ji Hyun; Nam, Ryoung Hee; Seok, Yeong-Jae; Kim, Yeon-Ran; Kim, Joo Sung; Kim, Jung Mogg; Kim, Jung Min; Lee, Dong Ho; Jung, Hyun Chae

2016-10-01

Little is known about the role of gastric microbiota except for Helicobacter pylori (HP) in human health and disease. We compared the differences of human gastric microbiota according to gastric cancer or control and HP infection status and assessed the role of bacteria other than HP. Gastric microbiota of 63 antral mucosal and 18 corpus mucosal samples were analyzed by bar-coded 454 pyrosequencing of the 16S rRNA gene. Antral samples were divided into four subgroups based on HP positivity in pyrosequencing and the presence of cancer. The analysis was focused on bacteria other than HP, especially nitrosating or nitrate-reducing bacteria (NB). The changes of NB in antral mucosa of 16 subjects were followed up. The number of NB other than HP (non-HP-NB) was two times higher in the cancer groups than in the control groups, but it did not reach statistical significance. The number of non-HP-NB tends to increase over time, but this phenomenon was prevented by HP eradication in the HP-positive control group, but not in the HP-positive cancer group. We could not find the significant role of bacteria other than HP in the gastric carcinogenesis. © 2016 John Wiley & Sons Ltd.

Denaturing gradient gel electrophoresis and barcoded pyrosequencing reveal unprecedented archaeal diversity in mangrove sediment and rhizosphere samples.

PubMed

Pires, Ana C C; Cleary, Daniel F R; Almeida, Adelaide; Cunha, Angela; Dealtry, Simone; Mendonça-Hagler, Leda C S; Smalla, Kornelia; Gomes, Newton C M

2012-08-01

Mangroves are complex ecosystems that regulate nutrient and sediment fluxes to the open sea. The importance of bacteria and fungi in regulating nutrient cycles has led to an interest in their diversity and composition in mangroves. However, very few studies have assessed Archaea in mangroves, and virtually nothing is known about whether mangrove rhizospheres affect archaeal diversity and composition. Here, we studied the diversity and composition of Archaea in mangrove bulk sediment and the rhizospheres of two mangrove trees, Rhizophora mangle and Laguncularia racemosa, using denaturing gradient gel electrophoresis (DGGE) and pyrosequencing of archaeal 16S rRNA genes with a nested-amplification approach. DGGE profiles revealed significant structural differences between bulk sediment and rhizosphere samples, suggesting that roots of both mangrove species influence the sediment archaeal community. Nearly all of the detected sequences obtained with pyrosequencing were identified as Archaea, but most were unclassified at the level of phylum or below. Archaeal richness was, furthermore, the highest in the L. racemosa rhizosphere, intermediate in bulk sediment, and the lowest in the R. mangle rhizosphere. This study shows that rhizosphere microhabitats of R. mangle and L. racemosa, common plants in subtropical mangroves located in Rio de Janeiro, Brazil, hosted distinct archaeal assemblages.

Denaturing Gradient Gel Electrophoresis and Barcoded Pyrosequencing Reveal Unprecedented Archaeal Diversity in Mangrove Sediment and Rhizosphere Samples

PubMed Central

Pires, Ana C. C.; Cleary, Daniel F. R.; Almeida, Adelaide; Cunha, Ângela; Dealtry, Simone; Mendonça-Hagler, Leda C. S.; Smalla, Kornelia

2012-01-01

Mangroves are complex ecosystems that regulate nutrient and sediment fluxes to the open sea. The importance of bacteria and fungi in regulating nutrient cycles has led to an interest in their diversity and composition in mangroves. However, very few studies have assessed Archaea in mangroves, and virtually nothing is known about whether mangrove rhizospheres affect archaeal diversity and composition. Here, we studied the diversity and composition of Archaea in mangrove bulk sediment and the rhizospheres of two mangrove trees, Rhizophora mangle and Laguncularia racemosa, using denaturing gradient gel electrophoresis (DGGE) and pyrosequencing of archaeal 16S rRNA genes with a nested-amplification approach. DGGE profiles revealed significant structural differences between bulk sediment and rhizosphere samples, suggesting that roots of both mangrove species influence the sediment archaeal community. Nearly all of the detected sequences obtained with pyrosequencing were identified as Archaea, but most were unclassified at the level of phylum or below. Archaeal richness was, furthermore, the highest in the L. racemosa rhizosphere, intermediate in bulk sediment, and the lowest in the R. mangle rhizosphere. This study shows that rhizosphere microhabitats of R. mangle and L. racemosa, common plants in subtropical mangroves located in Rio de Janeiro, Brazil, hosted distinct archaeal assemblages. PMID:22660713

Pyrosequencing reveals diverse microbial community associated with the zoanthid Palythoa australiae from the South China Sea.

PubMed

Sun, Wei; Zhang, Fengli; He, Liming; Li, Zhiyong

2014-05-01

Diverse sessile organisms inhabit the coral reef ecosystems, including corals, sponges, and sea anemones. In the past decades, scleractinian corals (Cnidaria, Anthozoa, Scleractinia) and their associated microorganisms have attracted much attention. Zoanthids (Cnidaria, Anthozoa, Zoanthidea) are commonly found in coral reefs. However, little is known about the community structure of zoanthid-associated microbiota. In this study, the microbial community associated with the zoanthid Palythoa australiae in the South China Sea was investigated by 454 pyrosequencing. As a result, 2,353 bacterial, 583 archaeal, and 36 eukaryotic microbial ribotypes were detected, respectively. A total of 22 bacterial phyla (16 formally described phyla and six candidate phyla) were recovered. Proteobacteria was the most abundant group, followed by Chloroflexi and Actinobacteria. High-abundance Rhizobiales and diverse Chloroflexi were observed in the bacterial community. The archaeal population was composed of Crenarchaeota and Euryarchaeota, with Marine Group I as the dominant lineage. In particular, Candidatus Nitrosopumilus dominated the archaeal community. Besides bacteria and archaea, the zoanthid harbored eukaryotic microorganisms including fungi and algae though their diversity was very low. This study provided the first insights into the microbial community associated with P. australiae by 454 pyrosequencing, consequently laid a basis for the understanding of the association of P. australiae-microbes symbioses.

Evaluation of two real time PCR assays for the detection of bacterial DNA in amniotic fluid.

PubMed

Girón de Velasco-Sada, Patricia; Falces-Romero, Iker; Quiles-Melero, Inmaculada; García-Perea, Adela; Mingorance, Jesús

2018-01-01

The aim of this study was to evaluate two non-commercial Real-Time PCR assays for the detection of microorganisms in amniotic fluid followed by identification by pyrosequencing. We collected 126 amniotic fluids from 2010 to 2015 for the evaluation of two Real-Time PCR assays for detection of bacterial DNA in amniotic fluid (16S Universal PCR and Ureaplasma spp. specific PCR). The method was developed in the Department of Microbiology of the University Hospital La Paz. Thirty-seven samples (29.3%) were positive by PCR/pyrosequencing and/or culture, 4 of them were mixed cultures with Ureaplasma urealyticum. The Universal 16S Real-Time PCR was compared with the standard culture (81.8% sensitivity, 97.4% specificity, 75% positive predictive value, 98% negative predictive value). The Ureaplasma spp. specific Real-Time PCR was compared with the Ureaplasma/Mycoplasma specific culture (92.3% sensitivity, 89.4% specificity, 50% positive predictive value, 99% negative predictive value) with statistically significant difference (p=0.005). Ureaplasma spp. PCR shows a rapid response time (5h from DNA extraction until pyrosequencing) when comparing with culture (48h). So, the response time of bacteriological diagnosis in suspected chorioamnionitis is reduced. Copyright © 2017 Elsevier B.V. All rights reserved.

Deep-branching Novel Lineages and High Diversity of Haptophytes in the Skagerrak (Norway) Uncovered by 454 Pyrosequencing

PubMed Central

Egge, Elianne S; Eikrem, Wenche; Edvardsen, Bente

2015-01-01

Microalgae in the division Haptophyta may be difficult to identify to species by microscopy because they are small and fragile. Here, we used high-throughput sequencing to explore the diversity of haptophytes in outer Oslofjorden, Skagerrak, and supplemented this with electron microscopy. Nano- and picoplanktonic subsurface samples were collected monthly for 2 yr, and the haptophytes were targeted by amplification of RNA/cDNA with Haptophyta-specific 18S ribosomal DNA V4 primers. Pyrosequencing revealed higher species richness of haptophytes than previously observed in the Skagerrak by microscopy. From ca. 400,000 reads we obtained 156 haptophyte operational taxonomic units (OTUs) after rigorous filtering and 99.5% clustering. The majority (84%) of the OTUs matched environmental sequences not linked to a morphological species, most of which were affiliated with the order Prymnesiales. Phylogenetic analyses including Oslofjorden OTUs and available cultured and environmental haptophyte sequences showed that several of the OTUs matched sequences forming deep-branching lineages, potentially representing novel haptophyte classes. Pyrosequencing also retrieved cultured species not previously reported by microscopy in the Skagerrak. Electron microscopy revealed species not yet genetically characterised and some potentially novel taxa. This study contributes to linking genotype to phenotype within this ubiquitous and ecologically important protist group, and reveals great, unknown diversity. PMID:25099994

Microbial Nitrogen Cycling Associated with the Early Diagenesis of Organic Matter in Subseafloor Sediments

NASA Astrophysics Data System (ADS)

Zhao, R.

2015-12-01

The early diagenesis of organic matter is the major energy source of marine sedimentary biosphere and thus controls its population size; however, the vertical distribution of any functional groups along with the diagenesis of organic matter is remained unclear, especially for those microbes involved in nitrogen transformation which serve as a major control on the nitrogen flux between reservoirs. Here we investigated the vertical distributions of various functional groups in five sediment cores retrieved from Arctic Mid-Ocean Ridge (AMOR), with emphasis on the nitrifiers, denitrifiers and anaerobic ammonium oxidizing bacteria (anammox). We observed the clear geochemical zonation associated with organic matter diagenesis in the sediments based on the pore water profiles of oxygen, nitrate, ammonium, manganese and sulfate, with distinct geochemical transition zones at the boundaries of geochemical zones, including oxic-anoxic transition zone (OATZ) and nitrate-manganese reduction zone (NMTZ). Nitrate was produced in surface oxygenated sediments and nitrate consumption mainly took place at the NMTZ, splitted between re-oxidation of ammonium and manganese (II). Abundances of ammonia oxidizers, nitrite oxidizers, and denitrifiers, estimated through quantitative PCR targeting their respective functional genes, generally decrease with depth, but constantly elevated around the OATZ, NMTZ, and manganese-reduction zone as well. Anammox bacteria were only detected around the NMTZ where both nitrate/nitrite and ammonium are available. These depth profiles of functional groups were also confirmed by the community structure profiling by prokaryotic 16S rRNA gene tag pyrosequencing. Cell-specific rates of nitrification and denitrification, calculated from the bulk net reaction rates divided by functional group abundances, were similar to those values from oligotrophic sediments like North Pond and thus suggested that nitrifiers and denitirifiers populations were in maintenance state. This study illustrated the microbial nitrogen transformation accompanying the early diagenesis of organic matter in marine sediments, which scenario might be occurring in a wide range of stratified environments on Earth.

Comparative metagenomics of microbial communities inhabiting deep-sea hydrothermal vent chimneys with contrasting chemistries

PubMed Central

Xie, Wei; Wang, Fengping; Guo, Lei; Chen, Zeling; Sievert, Stefan M; Meng, Jun; Huang, Guangrui; Li, Yuxin; Yan, Qingyu; Wu, Shan; Wang, Xin; Chen, Shangwu; He, Guangyuan; Xiao, Xiang; Xu, Anlong

2011-01-01

Deep-sea hydrothermal vent chimneys harbor a high diversity of largely unknown microorganisms. Although the phylogenetic diversity of these microorganisms has been described previously, the adaptation and metabolic potential of the microbial communities is only beginning to be revealed. A pyrosequencing approach was used to directly obtain sequences from a fosmid library constructed from a black smoker chimney 4143-1 in the Mothra hydrothermal vent field at the Juan de Fuca Ridge. A total of 308 034 reads with an average sequence length of 227 bp were generated. Comparative genomic analyses of metagenomes from a variety of environments by two-way clustering of samples and functional gene categories demonstrated that the 4143-1 metagenome clustered most closely with that from a carbonate chimney from Lost City. Both are highly enriched in genes for mismatch repair and homologous recombination, suggesting that the microbial communities have evolved extensive DNA repair systems to cope with the extreme conditions that have potential deleterious effects on the genomes. As previously reported for the Lost City microbiome, the metagenome of chimney 4143-1 exhibited a high proportion of transposases, implying that horizontal gene transfer may be a common occurrence in the deep-sea vent chimney biosphere. In addition, genes for chemotaxis and flagellar assembly were highly enriched in the chimney metagenomes, reflecting the adaptation of the organisms to the highly dynamic conditions present within the chimney walls. Reconstruction of the metabolic pathways revealed that the microbial community in the wall of chimney 4143-1 was mainly fueled by sulfur oxidation, putatively coupled to nitrate reduction to perform inorganic carbon fixation through the Calvin–Benson–Bassham cycle. On the basis of the genomic organization of the key genes of the carbon fixation and sulfur oxidation pathways contained in the large genomic fragments, both obligate and facultative autotrophs appear to be present and contribute to biomass production. PMID:20927138

Toward an understanding of global change: Initial priorities for US contributions to the International Geosphere-Biosphere Program

NASA Technical Reports Server (NTRS)

1988-01-01

A limited number of high-priority research initiatives are recommended for early implementation as part of the U.S. contribution to the preparatory phase of the International Geosphere-Biosphere Program. The recommendations are based on the committee's analysis of the most critical gaps in the scientific knowledge needed to understand the changes that are occurring in the earth system not being addressed by existing programs. The report articulates a number of important key issues and interactions that characterize global change in the geosphere-biosphere system on time scales of decades to centuries; identifies the knowledge that is the most urgently needed to improve understanding of those issues and interactions; and formulates initial priorities for initial U.S. contributions to the IGBP, recognizing the contributions of other ongoing and proposed programs.

A modern earth narrative: What will be the fate of the biosphere?

USGS Publications Warehouse

Williams, R.S.

2000-01-01

The modern Earth Narrative is the scientific description of the natural and human history of the Earth, and it is based on two fundamental concepts: Deep (or Geologic) Time and Biological Evolution. Changes in the Earth's biosphere and geosphere are discussed from the perspective of natural variability and impacts of the rapid increase in the human population. The failure of humans to comprehend and understand the Earth Narrative, especially the place of humans in it, presages dire consequences for the Earth's biosphere. The actions humans take, individually and collectively, during the 21st century will likely determine the fate of many millions of species, including our own. It is argued that we must quickly establish an Earth System-based conservation ethic that has the objective of complete preservation of the Earth's biotic inheritance. Published by Elsevier Science Ltd.

Global patterns and controls of soil organic carbon dynamics as simulated by multiple terrestrial biosphere models: Current status and future directions.

PubMed

Tian, Hanqin; Lu, Chaoqun; Yang, Jia; Banger, Kamaljit; Huntzinger, Deborah N; Schwalm, Christopher R; Michalak, Anna M; Cook, Robert; Ciais, Philippe; Hayes, Daniel; Huang, Maoyi; Ito, Akihiko; Jain, Atul K; Lei, Huimin; Mao, Jiafu; Pan, Shufen; Post, Wilfred M; Peng, Shushi; Poulter, Benjamin; Ren, Wei; Ricciuto, Daniel; Schaefer, Kevin; Shi, Xiaoying; Tao, Bo; Wang, Weile; Wei, Yaxing; Yang, Qichun; Zhang, Bowen; Zeng, Ning

2015-06-01

Soil is the largest organic carbon (C) pool of terrestrial ecosystems, and C loss from soil accounts for a large proportion of land-atmosphere C exchange. Therefore, a small change in soil organic C (SOC) can affect atmospheric carbon dioxide (CO 2 ) concentration and climate change. In the past decades, a wide variety of studies have been conducted to quantify global SOC stocks and soil C exchange with the atmosphere through site measurements, inventories, and empirical/process-based modeling. However, these estimates are highly uncertain, and identifying major driving forces controlling soil C dynamics remains a key research challenge. This study has compiled century-long (1901-2010) estimates of SOC storage and heterotrophic respiration (Rh) from 10 terrestrial biosphere models (TBMs) in the Multi-scale Synthesis and Terrestrial Model Intercomparison Project and two observation-based data sets. The 10 TBM ensemble shows that global SOC estimate ranges from 425 to 2111 Pg C (1 Pg = 10 15  g) with a median value of 1158 Pg C in 2010. The models estimate a broad range of Rh from 35 to 69 Pg C yr -1 with a median value of 51 Pg C yr -1 during 2001-2010. The largest uncertainty in SOC stocks exists in the 40-65°N latitude whereas the largest cross-model divergence in Rh are in the tropics. The modeled SOC change during 1901-2010 ranges from -70 Pg C to 86 Pg C, but in some models the SOC change has a different sign from the change of total C stock, implying very different contribution of vegetation and soil pools in determining the terrestrial C budget among models. The model ensemble-estimated mean residence time of SOC shows a reduction of 3.4 years over the past century, which accelerate C cycling through the land biosphere. All the models agreed that climate and land use changes decreased SOC stocks, while elevated atmospheric CO 2 and nitrogen deposition over intact ecosystems increased SOC stocks-even though the responses varied significantly among models. Model representations of temperature and moisture sensitivity, nutrient limitation, and land use partially explain the divergent estimates of global SOC stocks and soil C fluxes in this study. In addition, a major source of systematic error in model estimations relates to nonmodeled SOC storage in wetlands and peatlands, as well as to old C storage in deep soil layers.

Global patterns and controls of soil organic carbon dynamics as simulated by multiple terrestrial biosphere models: Current status and future directions

DOE PAGES

Tian, Hanqin; Lu, Chaoqun; Yang, Jia; ...

2015-06-05

Soil is the largest organic carbon (C) pool of terrestrial ecosystems, and C loss from soil accounts for a large proportion of land-atmosphere C exchange. Therefore, a small change in soil organic C (SOC) can affect atmospheric carbon dioxide (CO₂) concentration and climate change. In the past decades, a wide variety of studies have been conducted to quantify global SOC stocks and soil C exchange with the atmosphere through site measurements, inventories, and empirical/process-based modeling. However, these estimates are highly uncertain, and identifying major driving forces controlling soil C dynamics remains a key research challenge. This study has compiled century-longmore » (1901–2010) estimates of SOC storage and heterotrophic respiration (Rh) from 10 terrestrial biosphere models (TBMs) in the Multi-scale Synthesis and Terrestrial Model Intercomparison Project and two observation-based data sets. The 10 TBM ensemble shows that global SOC estimate ranges from 425 to 2111 Pg C (1 Pg = 10¹⁵ g) with a median value of 1158 Pg C in 2010. The models estimate a broad range of Rh from 35 to 69 Pg C yr⁻¹ with a median value of 51 Pg C yr⁻¹ during 2001–2010. The largest uncertainty in SOC stocks exists in the 40–65°N latitude whereas the largest cross-model divergence in Rh are in the tropics. The modeled SOC change during 1901–2010 ranges from –70 Pg C to 86 Pg C, but in some models the SOC change has a different sign from the change of total C stock, implying very different contribution of vegetation and soil pools in determining the terrestrial C budget among models. The model ensemble-estimated mean residence time of SOC shows a reduction of 3.4 years over the past century, which accelerate C cycling through the land biosphere. All the models agreed that climate and land use changes decreased SOC stocks, while elevated atmospheric CO₂ and nitrogen deposition over intact ecosystems increased SOC stocks—even though the responses varied significantly among models. Model representations of temperature and moisture sensitivity, nutrient limitation, and land use partially explain the divergent estimates of global SOC stocks and soil C fluxes in this study. In addition, a major source of systematic error in model estimations relates to nonmodeled SOC storage in wetlands and peatlands, as well as to old C storage in deep soil layers.« less

High-Throughput Sequence Analysis of Turbot (Scophthalmus maximus) Transcriptome Using 454-Pyrosequencing for the Discovery of Antiviral Immune Genes

PubMed Central

Pereiro, Patricia; Balseiro, Pablo; Romero, Alejandro; Dios, Sonia; Forn-Cuni, Gabriel; Fuste, Berta; Planas, Josep V.; Beltran, Sergi; Novoa, Beatriz; Figueras, Antonio

2012-01-01

Background Turbot (Scophthalmus maximus L.) is an important aquacultural resource both in Europe and Asia. However, there is little information on gene sequences available in public databases. Currently, one of the main problems affecting the culture of this flatfish is mortality due to several pathogens, especially viral diseases which are not treatable. In order to identify new genes involved in immune defense, we conducted 454-pyrosequencing of the turbot transcriptome after different immune stimulations. Methodology/Principal Findings Turbot were injected with viral stimuli to increase the expression level of immune-related genes. High-throughput deep sequencing using 454-pyrosequencing technology yielded 915,256 high-quality reads. These sequences were assembled into 55,404 contigs that were subjected to annotation steps. Intriguingly, 55.16% of the deduced protein was not significantly similar to any sequences in the databases used for the annotation and only 0.85% of the BLASTx top-hits matched S. maximus protein sequences. This relatively low level of annotation is possibly due to the limited information for this specie and other flatfish in the database. These results suggest the identification of a large number of new genes in turbot and in fish in general. A more detailed analysis showed the presence of putative members of several innate and specific immune pathways. Conclusions/Significance To our knowledge, this study is the first transcriptome analysis using 454-pyrosequencing for turbot. Previously, there were only 12,471 EST and less of 1,500 nucleotide sequences for S. maximus in NCBI database. Our results provide a rich source of data (55,404 contigs and 181,845 singletons) for discovering and identifying new genes, which will serve as a basis for microarray construction, gene expression characterization and for identification of genetic markers to be used in several applications. Immune stimulation in turbot was very effective, obtaining an enormous variety of sequences belonging to genes involved in the defense mechanisms. PMID:22629298

Transcriptome Exploration in Leymus chinensis under Saline-Alkaline Treatment Using 454 Pyrosequencing

PubMed Central

Sun, Yepeng; Wang, Fawei; Wang, Nan; Dong, Yuanyuan; Liu, Qi; Zhao, Lei; Chen, Huan; Liu, Weican; Yin, Hailong; Zhang, Xiaomei; Yuan, Yanxi; Li, Haiyan

2013-01-01

Background Leymus chinensis (Trin.) Tzvel. is a high saline-alkaline tolerant forage grass genus of the tribe Gramineae family, which also plays an important role in protection of natural environment. To date, little is known about the saline-alkaline tolerance of L. chinensis on the molecular level. To better understand the molecular mechanism of saline-alkaline tolerance in L. chinensis, 454 pyrosequencing was used for the transcriptome study. Results We used Roche-454 massive parallel pyrosequencing technology to sequence two different cDNA libraries that were built from the two samples of control and under saline-alkaline treatment (optimal stress concentration-Hoagland solution with 100 mM NaCl and 200 mM NaHCO3). A total of 363,734 reads in control group and 526,267 reads in treatment group with an average length of 489 bp and 493 bp were obtained, respectively. The reads were assembled into 104,105 unigenes with MIRA sequence assemable software, among which, 73,665 unigenes were in control group, 88,016 unigenes in treatment group and 57,576 unigenes in both groups. According to the comparative expression analysis between the two groups with the threshold of “log2 Ratio ≥1”, there were 36,497 up-regulated unegenes and 18,218 down-regulated unigenes predicted to be the differentially expressed genes. After gene annotation and pathway enrichment analysis, most of them were involved in stress and tolerant function, signal transduction, energy production and conversion, and inorganic ion transport. Furthermore, 16 of these differentially expressed genes were selected for real-time PCR validation, and they were successfully confirmed with the results of 454 pyrosequencing. Conclusions This work is the first time to study the transcriptome of L. chinensis under saline-alkaline treatment based on the 454-FLX massively parallel DNA sequencing platform. It also deepened studies on molecular mechanisms of saline-alkaline in L. chinensis, and constituted a database for future studies. PMID:23365637

Detection of KRAS G12D in colorectal cancer stool by droplet digital PCR

PubMed Central

Olmedillas-López, Susana; Lévano-Linares, Dennis César; Alexandre, Carmen Laura Aúz; Vega-Clemente, Luz; Sánchez, Edurne León; Villagrasa, Alejandro; Ruíz-Tovar, Jaime; García-Arranz, Mariano; García-Olmo, Damián

2017-01-01

AIM To assess KRAS G12D mutation detection by droplet digital PCR (ddPCR) in stool-derived DNA from colorectal cancer (CRC) patients. METHODS In this study, tumor tissue and stool samples were collected from 70 patients with stage I-IV CRC diagnosed by preoperative biopsy. KRAS mutational status was determined by pyrosequencing analysis of DNA obtained from formalin-fixed paraffin-embedded (FFPE) tumor tissues. The KRAS G12D mutation was then analyzed by ddPCR in FFPE tumors and stool-derived DNA from patients with this point mutation. Wild-type (WT) tumors, as determined by pyrosequencing, were included as controls; analysis of FFPE tissue and stool-derived DNA by ddPCR was performed for these patients as well. RESULTS Among the total 70 patients included, KRAS mutations were detected by pyrosequencing in 32 (45.71%), whereas 38 (54.29%) had WT tumors. The frequency of KRAS mutations was higher in left-sided tumors (11 located in the right colon, 15 in the left, and 6 in the rectum). The predominant point mutation was KRAS G12D (14.29%, n = 10), which was more frequent in early-stage tumors (I-IIA, n = 7). In agreement with pyrosequencing results, the KRAS G12D mutation was detected by ddPCR in FFPE tumor-derived DNA, and only a residual number of mutated copies was found in WT controls. The KRAS G12D mutation was also detected in stool-derived DNA in 80% of all fecal samples from CRC patients with this point mutation. CONCLUSION ddPCR is a reliable and sensitive method to analyze KRAS G12D mutation in stool-derived DNA from CRC patients, especially at early stages. This non-invasive approach is potentially applicable to other relevant biomarkers for CRC management. PMID:29093617

Evaluation of six primer pairs targeting the nuclear rRNA operon for characterization of arbuscular mycorrhizal fungal (AMF) communities using 454 pyrosequencing.

PubMed

Van Geel, Maarten; Busschaert, Pieter; Honnay, Olivier; Lievens, Bart

2014-11-01

In the last few years, 454 pyrosequencing-based analysis of arbuscular mycorrhizal fungal (AMF; Glomeromycota) communities has tremendously increased our knowledge of the distribution and diversity of AMF. Nonetheless, comparing results between different studies is difficult, as different target genes (or regions thereof) and primer combinations, with potentially dissimilar specificities and efficacies, are being utilized. In this study we evaluated six primer pairs that have previously been used in AMF studies (NS31-AM1, AMV4.5NF-AMDGR, AML1-AML2, NS31-AML2, FLR3-LSUmBr and Glo454-NDL22) for their use in 454 pyrosequencing based on both an in silico approach and 454 pyrosequencing of AMF communities from apple tree roots. Primers were evaluated in terms of (i) in silico coverage of Glomeromycota fungi, (ii) the number of high-quality sequences obtained, (iii) selectivity for AMF species, (iv) reproducibility and (v) ability to accurately describe AMF communities. We show that primer pairs AMV4.5NF-AMDGR, AML1-AML2 and NS31-AML2 outperformed the other tested primer pairs in terms of number of Glomeromycota reads (AMF specificity and coverage). Additionally, these primer pairs were found to have no or only few mismatches to AMF sequences and were able to consistently describe AMF communities from apple roots. However, whereas most high-quality AMF sequences were obtained for AMV4.5NF-AMDGR, our results also suggest that this primer pair favored amplification of Glomeraceae sequences at the expense of Ambisporaceae, Claroideoglomeraceae and Paraglomeraceae sequences. Furthermore, we demonstrate the complementary specificity of AMV4.5NF-AMDGR with AML1-AML2, and of AMV4.5NF-AMDGR with NS31-AML2, making these primer combinations highly suitable for tandem use in covering the diversity of AMF communities. Copyright © 2014 Elsevier B.V. All rights reserved.

Pyrosequencing the Bemisia tabaci Transcriptome Reveals a Highly Diverse Bacterial Community and a Robust System for Insecticide Resistance

PubMed Central

Wu, Qing-jun; Wang, Shao-li; Yang, Xin; Yang, Ni-na; Li, Ru-mei; Jiao, Xiao-guo; Pan, Hui-peng; Liu, Bai-ming; Su, Qi; Xu, Bao-yun; Hu, Song-nian; Zhou, Xu-guo; Zhang, You-jun

2012-01-01

Background Bemisia tabaci (Gennadius) is a phloem-feeding insect poised to become one of the major insect pests in open field and greenhouse production systems throughout the world. The high level of resistance to insecticides is a main factor that hinders continued use of insecticides for suppression of B. tabaci. Despite its prevalence, little is known about B. tabaci at the genome level. To fill this gap, an invasive B. tabaci B biotype was subjected to pyrosequencing-based transcriptome analysis to identify genes and gene networks putatively involved in various physiological and toxicological processes. Methodology and Principal Findings Using Roche 454 pyrosequencing, 857,205 reads containing approximately 340 megabases were obtained from the B. tabaci transcriptome. De novo assembly generated 178,669 unigenes including 30,980 from insects, 17,881 from bacteria, and 129,808 from the nohit. A total of 50,835 (28.45%) unigenes showed similarity to the non-redundant database in GenBank with a cut-off E-value of 10–5. Among them, 40,611 unigenes were assigned to one or more GO terms and 6,917 unigenes were assigned to 288 known pathways. De novo metatranscriptome analysis revealed highly diverse bacterial symbionts in B. tabaci, and demonstrated the host-symbiont cooperation in amino acid production. In-depth transcriptome analysis indentified putative molecular markers, and genes potentially involved in insecticide resistance and nutrient digestion. The utility of this transcriptome was validated by a thiamethoxam resistance study, in which annotated cytochrome P450 genes were significantly overexpressed in the resistant B. tabaci in comparison to its susceptible counterparts. Conclusions This transcriptome/metatranscriptome analysis sheds light on the molecular understanding of symbiosis and insecticide resistance in an agriculturally important phloem-feeding insect pest, and lays the foundation for future functional genomics research of the B. tabaci complex. Moreover, current pyrosequencing effort greatly enriched the existing whitefly EST database, and makes RNAseq a viable option for future genomic analysis. PMID:22558125

Shotgun Pyrosequencing Metagenomic Analyses of Dusts from Swine Confinement and Grain Facilities

PubMed Central

Boissy, Robert J.; Romberger, Debra J.; Roughead, William A.; Weissenburger-Moser, Lisa; Poole, Jill A.; LeVan, Tricia D.

2014-01-01

Inhalation of agricultural dusts causes inflammatory reactions and symptoms such as headache, fever, and malaise, which can progress to chronic airway inflammation and associated diseases, e.g. asthma, chronic bronchitis, chronic obstructive pulmonary disease, and hypersensitivity pneumonitis. Although in many agricultural environments feed particles are the major constituent of these dusts, the inflammatory responses that they provoke are likely attributable to particle-associated bacteria, archaebacteria, fungi, and viruses. In this study, we performed shotgun pyrosequencing metagenomic analyses of DNA from dusts from swine confinement facilities or grain elevators, with comparisons to dusts from pet-free households. DNA sequence alignment showed that 19% or 62% of shotgun pyrosequencing metagenomic DNA sequence reads from swine facility or household dusts, respectively, were of swine or human origin, respectively. In contrast only 2% of such reads from grain elevator dust were of mammalian origin. These metagenomic shotgun reads of mammalian origin were excluded from our analyses of agricultural dust microbiota. The ten most prevalent bacterial taxa identified in swine facility compared to grain elevator or household dust were comprised of 75%, 16%, and 42% gram-positive organisms, respectively. Four of the top five swine facility dust genera were assignable (Clostridium, Lactobacillus, Ruminococcus, and Eubacterium, ranging from 4% to 19% relative abundance). The relative abundances of these four genera were lower in dust from grain elevators or pet-free households. These analyses also highlighted the predominance in swine facility dust of Firmicutes (70%) at the phylum level, Clostridia (44%) at the Class level, and Clostridiales at the Order level (41%). In summary, shotgun pyrosequencing metagenomic analyses of agricultural dusts show that they differ qualitatively and quantitatively at the level of microbial taxa present, and that the bioinformatic analyses used for such studies must be carefully designed to avoid the potential contribution of non-microbial DNA, e.g. from resident mammals. PMID:24748147

Shotgun pyrosequencing metagenomic analyses of dusts from swine confinement and grain facilities.

PubMed

Boissy, Robert J; Romberger, Debra J; Roughead, William A; Weissenburger-Moser, Lisa; Poole, Jill A; LeVan, Tricia D

2014-01-01

Inhalation of agricultural dusts causes inflammatory reactions and symptoms such as headache, fever, and malaise, which can progress to chronic airway inflammation and associated diseases, e.g. asthma, chronic bronchitis, chronic obstructive pulmonary disease, and hypersensitivity pneumonitis. Although in many agricultural environments feed particles are the major constituent of these dusts, the inflammatory responses that they provoke are likely attributable to particle-associated bacteria, archaebacteria, fungi, and viruses. In this study, we performed shotgun pyrosequencing metagenomic analyses of DNA from dusts from swine confinement facilities or grain elevators, with comparisons to dusts from pet-free households. DNA sequence alignment showed that 19% or 62% of shotgun pyrosequencing metagenomic DNA sequence reads from swine facility or household dusts, respectively, were of swine or human origin, respectively. In contrast only 2% of such reads from grain elevator dust were of mammalian origin. These metagenomic shotgun reads of mammalian origin were excluded from our analyses of agricultural dust microbiota. The ten most prevalent bacterial taxa identified in swine facility compared to grain elevator or household dust were comprised of 75%, 16%, and 42% gram-positive organisms, respectively. Four of the top five swine facility dust genera were assignable (Clostridium, Lactobacillus, Ruminococcus, and Eubacterium, ranging from 4% to 19% relative abundance). The relative abundances of these four genera were lower in dust from grain elevators or pet-free households. These analyses also highlighted the predominance in swine facility dust of Firmicutes (70%) at the phylum level, Clostridia (44%) at the Class level, and Clostridiales at the Order level (41%). In summary, shotgun pyrosequencing metagenomic analyses of agricultural dusts show that they differ qualitatively and quantitatively at the level of microbial taxa present, and that the bioinformatic analyses used for such studies must be carefully designed to avoid the potential contribution of non-microbial DNA, e.g. from resident mammals.

Monitoring of microbial communities in anaerobic digestion sludge for biogas optimisation.

PubMed

Lim, Jun Wei; Ge, Tianshu; Tong, Yen Wah

2018-01-01

This study characterised and compared the microbial communities of anaerobic digestion (AD) sludge using three different methods - (1) Clone library; (2) Pyrosequencing; and (3) Terminal restriction fragment length polymorphism (T-RFLP). Although high-throughput sequencing techniques are becoming increasingly popular and affordable, the reliance of such techniques for frequent monitoring of microbial communities may be a financial burden for some. Furthermore, the depth of microbial analysis revealed by high-throughput sequencing may not be required for monitoring purposes. This study aims to develop a rapid, reliable and economical approach for the monitoring of microbial communities in AD sludge. A combined approach where genetic information of sequences from clone library was used to assign phylogeny to T-RFs determined experimentally was developed in this study. In order to assess the effectiveness of the combined approach, microbial communities determined by the combined approach was compared to that characterised by pyrosequencing. Results showed that both pyrosequencing and clone library methods determined the dominant bacteria phyla to be Proteobacteria, Firmicutes, Bacteroidetes, and Thermotogae. Both methods also found that sludge A and B were predominantly dominated by acetogenic methanogens followed by hydrogenotrophic methanogens. The number of OTUs detected by T-RFLP was significantly lesser than that detected by the clone library. In this study, T-RFLP analysis identified majority of the dominant species of the archaeal consortia. However, many of the more highly diverse bacteria consortia were missed. Nevertheless, the combined approach developed in this study where clone sequences from the clone library were used to assign phylogeny to T-RFs determined experimentally managed to accurately predict the same dominant microbial groups for both sludge A and sludge B, as compared to the pyrosequencing results. Results showed that the combined approach of clone library and T-RFLP accurately predicted the dominant microbial groups and thus is a reliable and more economical way to monitor the evolution of microbial systems in AD sludge. Copyright © 2017 Elsevier Ltd. All rights reserved.

Analysis of oral microbiota in children with dental caries by PCR-DGGE and barcoded pyrosequencing.

PubMed

Ling, Zongxin; Kong, Jianming; Jia, Peng; Wei, Chaochun; Wang, Yuezhu; Pan, Zhiwen; Huang, Wujing; Li, Lanjuan; Chen, Hui; Xiang, Charlie

2010-10-01

Oral microbiota plays a vital role in maintaining the homeostasis of oral cavity. Dental caries are among the most common oral diseases in children and pathogenic bacteria contribute to the development of the disease. However, the overall structure of bacterial communities in the oral cavity from children with dental caries has not been explored deeply heretofore. We used high-throughput barcoded pyrosequencing and PCR-denaturing gradient gel electrophoresis (DGGE) to examine bacterial diversity of oral microbiota in saliva and supragingival plaques from 60 children aged 3 to 6 years old with and without dental caries from China. The multiplex barcoded pyrosequencing was performed in a single run, with multiple samples tagged uniquely by multiplex identifiers. As PCR-DGGE analysis is a conventional molecular ecological approach, this analysis was also performed on the same samples and the results of both approaches were compared. A total of 186,787 high-quality sequences were obtained for evaluating bacterial diversity and 41,905 unique sequences represented all phylotypes. We found that the oral microbiota in children was far more diverse than previous studies reported, and more than 200 genera belonging to ten phyla were found in the oral cavity. The phylotypes in saliva and supragingival plaques were significantly different and could be divided into two distinct clusters (p < 0.05). The bacterial diversity in oral microbiome analyzed by PCR-DGGE and barcoded pyrosequencing was employed to cross validate the data sets. The genera of Streptococcus, Veillonella, Actinomyces, Granulicatella, Leptotrichia, and Thiomonas in plaques were significantly associated with dental caries (p < 0.05). The results showed that there was no one specific pathogen but rather pathogenic populations in plaque that significantly correlated with dental caries. The enormous diversity of oral microbiota allowed for a better understanding of oral microecosystem, and these pathogenic populations in plaque provide new insights into the etiology of dental caries and suggest new targets for interventions of the disease.

Evolving Methanococcoides burtonii archaeal Rubisco for improved photosynthesis and plant growth.

PubMed

Wilson, Robert H; Alonso, Hernan; Whitney, Spencer M

2016-03-01

In photosynthesis Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyses the often rate limiting CO2-fixation step in the Calvin cycle. This makes Rubisco both the gatekeeper for carbon entry into the biosphere and a target for functional improvement to enhance photosynthesis and plant growth. Encumbering the catalytic performance of Rubisco is its highly conserved, complex catalytic chemistry. Accordingly, traditional efforts to enhance Rubisco catalysis using protracted "trial and error" protein engineering approaches have met with limited success. Here we demonstrate the versatility of high throughput directed (laboratory) protein evolution for improving the carboxylation properties of a non-photosynthetic Rubisco from the archaea Methanococcoides burtonii. Using chloroplast transformation in the model plant Nicotiana tabacum (tobacco) we confirm the improved forms of M. burtonii Rubisco increased photosynthesis and growth relative to tobacco controls producing wild-type M. burtonii Rubisco. Our findings indicate continued directed evolution of archaeal Rubisco offers new potential for enhancing leaf photosynthesis and plant growth.

Intergovernmental Conference of Experts on the Scientific Basis for Rational Use and Conservation of the Resources of the Biosphere (Paris, France, September 4-13, 1968), Recommendations.

ERIC Educational Resources Information Center

United Nations Educational, Scientific, and Cultural Organization, Paris (France).

This document contains twenty recommendations put forth at the Intergovernmental Conference of Experts on the Scientific Basis for Rational Use and Conservation of the Resources of the Biosphere under the direction of UNESCO (United Nations Educational, Scientific and Cultural Organization). The conference was held at the UNESCO House, Paris,…

Studies of land-cover, land-use, and biophysical properties of vegetation in the Large Scale Biosphere Atmosphere experiment in Amazonia.

Treesearch

Dar A. Robertsa; Michael Keller; Joao Vianei Soares

2003-01-01

We summarize early research on land-cover, land-use, and biophysical properties of vegetation from the Large Scale Biosphere Atmosphere (LBA) experiment in Amazoˆnia. LBA is an international research program developed to evaluate regional function and to determine how land-use and climate modify biological, chemical and physical processes there. Remote sensing has...

Effects of primitive photosynthesis on Earth's early climate system

NASA Astrophysics Data System (ADS)

Ozaki, Kazumi; Tajika, Eiichi; Hong, Peng K.; Nakagawa, Yusuke; Reinhard, Christopher T.

2018-01-01

The evolution of different forms of photosynthetic life has profoundly altered the activity level of the biosphere, radically reshaping the composition of Earth's oceans and atmosphere over time. However, the mechanistic impacts of a primitive photosynthetic biosphere on Earth's early atmospheric chemistry and climate are poorly understood. Here, we use a global redox balance model to explore the biogeochemical and climatological effects of different forms of primitive photosynthesis. We find that a hybrid ecosystem of H2-based and Fe2+-based anoxygenic photoautotrophs—organisms that perform photosynthesis without producing oxygen—gives rise to a strong nonlinear amplification of Earth's methane (CH4) cycle, and would thus have represented a critical component of Earth's early climate system before the advent of oxygenic photosynthesis. Using a Monte Carlo approach, we find that a hybrid photosynthetic biosphere widens the range of geochemical conditions that allow for warm climate states well beyond either of these metabolic processes acting in isolation. Our results imply that the Earth's early climate was governed by a novel and poorly explored set of regulatory feedbacks linking the anoxic biosphere and the coupled H, C and Fe cycles. We suggest that similar processes should be considered when assessing the potential for sustained habitability on Earth-like planets with reducing atmospheres.

Intraterrestrial life in igneous ocean crust: advances, technologies, and the future (Invited)

NASA Astrophysics Data System (ADS)

Edwards, K. J.; Wheat, C. G.

2010-12-01

The “next frontier” of scientific investigation in the deep sub-seafloor microbial biosphere lies in a realm that has been a completely unexplored until just the past decade: the igneous oceanic crust. Problems that have hampered exploration of the “hard rock” marine deep biosphere have revolved around sample access (hard rock drilling is technologically complex), contamination (a major hurdle), momentum (why take on this challenge when the relatively “easier” marine muds also have been a frontier) and suspicion that microbes in more readily accessed using (simpler) non-drilling technologies - like vents - are truly are endemic of subsurface clades/activities. Since the late 1990’s, however, technologies and resultant studies on microbes in the igneous ocean crust deep biosphere have risen sharply, and offer a new and distinct view on this biome. Moreover, microbiologists are now taking leading roles in technological developments that are critically required to address this biosphere - interfacing and collaborating closely with engineers, genomic biologists, geologists, seismologists, and geochemists to accomplish logistically complex and long-term studies that bring observatory research to this deep realm. The future of this field for the least decade is rich - opportunities abound for microbiologists to play new roles in how we study microbiology in the deep subsurface in an oceanographic and Earth system science perspective.

Searching for a shadow biosphere on Earth as a test of the 'cosmic imperative'.

PubMed

Davies, P C W

2011-02-13

Estimates for the number of communicating civilizations in the galaxy, based on the so-called Drake equation, are meaningless without a plausible estimate for the probability that life will emerge on an Earth-like planet. In the absence of a theory of the origin of life, that number can be anywhere from 0 to 1. Distinguished scientists have been known to argue that life on Earth is a freak accident, unique in the observable universe and, conversely, that life is almost bound to arise in the course of time, given Earth-like conditions. De Duve, adopting the latter position, coined the phrase that 'life is a cosmic imperative'. De Duve's position would be immediately verified if we were to discover a second sample of life that we could be sure arose from scratch independently of known life. Given the current absence of evidence for life beyond Earth, the best way to test the hypothesis of the cosmic imperative is to see whether terrestrial life began more than once. If it did, it is possible that descendants of a second genesis might be extant, forming a sort of 'shadow biosphere' existing alongside, or perhaps interpenetrating, the known biosphere. I outline a strategy to detect the existence of such a shadow biosphere.

Life in a dark biosphere: a review of circadian physiology in "arrhythmic" environments.

PubMed

Beale, Andrew David; Whitmore, David; Moran, Damian

2016-12-01

Most of the life with which humans interact is exposed to highly rhythmic and extremely predictable changes in illumination that occur with the daily events of sunrise and sunset. However, while the influence of the sun feels omnipotent to surface dwellers such as ourselves, life on earth is dominated, in terms of biomass, by organisms isolated from the direct effects of the sun. A limited understanding of what life is like away from the sun can be inferred from our knowledge of physiology and ecology in the light biosphere, but a full understanding can only be gained by studying animals from the dark biosphere, both in the laboratory and in their natural habitats. One of the least understood aspects of life in the dark biosphere is the rhythmicity of physiology and what it means to live in an environment of low or no rhythmicity. Here we describe methods that may be used to understand rhythmic physiology in the dark and summarise some of the studies of rhythmic physiology in "arrhythmic" environments, such as the poles, deep sea and caves. We review what can be understood about the adaptive value of rhythmic physiology on the Earth's surface from studies of animals from arrhythmic environments and what role a circadian clock may play in the dark.

Microbial metagenomes from three aquifers in the Fennoscandian shield terrestrial deep biosphere reveal metabolic partitioning among populations

PubMed Central

Wu, Xiaofen; Holmfeldt, Karin; Hubalek, Valerie; Lundin, Daniel; Åström, Mats; Bertilsson, Stefan; Dopson, Mark

2016-01-01

Microorganisms in the terrestrial deep biosphere host up to 20% of the earth's biomass and are suggested to be sustained by the gases hydrogen and carbon dioxide. A metagenome analysis of three deep subsurface water types of contrasting age (from <20 to several thousand years) and depth (171 to 448 m) revealed phylogenetically distinct microbial community subsets that either passed or were retained by a 0.22 μm filter. Such cells of <0.22 μm would have been overlooked in previous studies relying on membrane capture. Metagenomes from the three water types were used for reconstruction of 69 distinct microbial genomes, each with >86% coverage. The populations were dominated by Proteobacteria, Candidate divisions, unclassified archaea and unclassified bacteria. The estimated genome sizes of the <0.22 μm populations were generally smaller than their phylogenetically closest relatives, suggesting that small dimensions along with a reduced genome size may be adaptations to oligotrophy. Shallow ‘modern marine' water showed community members with a predominantly heterotrophic lifestyle. In contrast, the deeper, ‘old saline' water adhered more closely to the current paradigm of a hydrogen-driven deep biosphere. The data were finally used to create a combined metabolic model of the deep terrestrial biosphere microbial community. PMID:26484735

Microbial metagenomes from three aquifers in the Fennoscandian shield terrestrial deep biosphere reveal metabolic partitioning among populations.

PubMed

Wu, Xiaofen; Holmfeldt, Karin; Hubalek, Valerie; Lundin, Daniel; Åström, Mats; Bertilsson, Stefan; Dopson, Mark

2016-05-01

Microorganisms in the terrestrial deep biosphere host up to 20% of the earth's biomass and are suggested to be sustained by the gases hydrogen and carbon dioxide. A metagenome analysis of three deep subsurface water types of contrasting age (from <20 to several thousand years) and depth (171 to 448 m) revealed phylogenetically distinct microbial community subsets that either passed or were retained by a 0.22 μm filter. Such cells of <0.22 μm would have been overlooked in previous studies relying on membrane capture. Metagenomes from the three water types were used for reconstruction of 69 distinct microbial genomes, each with >86% coverage. The populations were dominated by Proteobacteria, Candidate divisions, unclassified archaea and unclassified bacteria. The estimated genome sizes of the <0.22 μm populations were generally smaller than their phylogenetically closest relatives, suggesting that small dimensions along with a reduced genome size may be adaptations to oligotrophy. Shallow 'modern marine' water showed community members with a predominantly heterotrophic lifestyle. In contrast, the deeper, 'old saline' water adhered more closely to the current paradigm of a hydrogen-driven deep biosphere. The data were finally used to create a combined metabolic model of the deep terrestrial biosphere microbial community.

Community structure of rare methanogenic archaea: insight from a single functional group

PubMed Central

Winkel, Matthias; Wagner, Dirk

2017-01-01

Abstract The rare biosphere, the low abundant microbial populations, is suggested to be a conserved way of microbial life. Here we conducted a molecular survey of rare methanogenic archaea in the environment targeting the mcrA gene in order to test if general concepts associated with the structure of the rare bacterial biosphere also apply to single functional groups. Similar to what is known about rare bacterial communities, the contribution of rare methanogens to the alpha diversity is much larger than to Bray-Curtis measures. Moreover, a similar core group of methanogens harbored by the abundant and rare communities suggests similar sources and environmental controls of both groups. Among the communities of different levels of rarity, the conditionally rare methanogenic taxa largely account for the overall community dynamics of the rare biosphere and likely enter the dominant community under favorable environmental conditions. In addition, we observed a positive correlation between the alpha diversity and the production of methane when the rare taxa were taken into account. This supports the concept that increasing microbial biodiversity enhances ecological function. The composition and environmental associations of the rare methanogenic biosphere allow us to conclude that rarity is a conserved way also for single functional groups. PMID:29029047

Development of an ELA-DRA gene typing method based on pyrosequencing technology.

PubMed

Díaz, S; Echeverría, M G; It, V; Posik, D M; Rogberg-Muñoz, A; Pena, N L; Peral-García, P; Vega-Pla, J L; Giovambattista, G

2008-11-01

The polymorphism of equine lymphocyte antigen (ELA) class II DRA gene had been detected by polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP) and reference strand-mediated conformation analysis. These methodologies allowed to identify 11 ELA-DRA exon 2 sequences, three of which are widely distributed among domestic horse breeds. Herein, we describe the development of a pyrosequencing-based method applicable to ELA-DRA typing, by screening samples from eight different horse breeds previously typed by PCR-SSCP. This sequence-based method would be useful in high-throughput genotyping of major histocompatibility complex genes in horses and other animal species, making this system interesting as a rapid screening method for animal genotyping of immune-related genes.

Interplay between solid Earth and biological evolution

NASA Astrophysics Data System (ADS)

Höning, Dennis; Spohn, Tilman

2017-04-01

Major shifts in Earth's evolution led to progressive adaptations of the biosphere. Particularly the emergence of continents permitted efficient use of solar energy. However, the widespread evolution of the biosphere fed back to the Earth system, often argued as a cause for the great oxidation event or as an important component in stabilizing Earth's climate. Furthermore, biologically enhanced weathering rates alter the flux of sediments in subduction zones, establishing a potential link to the deep interior. Stably bound water within subducting sediments not only enhances partial melting but further affects the mantle rheology. The mantle responds by enhancing its rates of convection, water outgassing, and subduction. How crucial is the emergence and evolution of life on Earth to these processes, and how would Earth have been evolved without the emergence of life? We here discuss concepts and present models addressing these questions and discuss the biosphere as a major component in evolving Earth system feedback cycles.

An extensive phase space for the potential martian biosphere.

PubMed

Jones, Eriita G; Lineweaver, Charles H; Clarke, Jonathan D

2011-12-01

We present a comprehensive model of martian pressure-temperature (P-T) phase space and compare it with that of Earth. Martian P-T conditions compatible with liquid water extend to a depth of ∼310 km. We use our phase space model of Mars and of terrestrial life to estimate the depths and extent of the water on Mars that is habitable for terrestrial life. We find an extensive overlap between inhabited terrestrial phase space and martian phase space. The lower martian surface temperatures and shallower martian geotherm suggest that, if there is a hot deep biosphere on Mars, it could extend 7 times deeper than the ∼5 km depth of the hot deep terrestrial biosphere in the crust inhabited by hyperthermophilic chemolithotrophs. This corresponds to ∼3.2% of the volume of present-day Mars being potentially habitable for terrestrial-like life.

The oxygen side of sulfate constrains global biospheric productivity in the mid-Phanerozoic

NASA Astrophysics Data System (ADS)

Wing, B. A.

2009-12-01

Recent work has read in the oxygen side of sulfate a record of CO2 concentrations and extreme environments on Neoproterozoic Earth [1, 2]. The connection between CO2 levels and total isotopic composition of mineral proxies for marine sulfate was motivated by an empirical correlation between CO2 concentrations and in 17O anomalies in samples of atmospheric gases preserved as bubbles in ice cores [3,4]. The 17O anomaly in atmospheric O2 has been interpreted to originate primarily from stratospheric photochemical cycles of O3, O2, and CO2 [3, 4]. Both CO2 and O3 form the 17O-enriched partners for the 17O-depleted O2 and, given a fixed atmospheric lifetime for O2, isotopic mass balance dictates that increasing CO2 levels will drive larger relative 17O deficits in O2. With a photochemically-calibrated relationship between the relative 17O anomaly in atmospheric O2 and atmospheric CO2 levels [1], the amount of CO2 in an ancient atmosphere can be directly estimated from the isotopic record of atmospheric O2 bound up in the oxygen side of sulfate. Although they are correlated, the relative 17O anomaly in atmospheric O2 is not only a function of atmospheric CO2 levels. Photosynthetic O2 is characterized by isotopically ‘normal’ oxygen sourced from the global hydrosphere [3]. Increased photosynthetic O2 production, therefore, dilutes the isotopic anomaly found in atmospheric O2. Measurements of the 17O anomaly in O2 from ice cores allow global changes in global biosphere productivity to be traced back to 60 ka ago [3, 4]. Applying similar analysis to recent datasets of the 17O anomaly in marine sulfates [1] and atmospheric CO2 levels [5] produces quantitative global biospheric productivity estimates for the time interval from 310 to 240 Ma ago. Although the time resolution is coarse, much of the mid-Phanerozoic was characterized by global biospheric productivity similar in magnitude to the average global biospheric productivity for the last 10 ka. Counterintuitively, this includes the global mid-Carboniferous biosphere. Global biospheric productivity at the end of the Permian, however, was apparently greatly enhanced relative to the rest of the mid-Phanerozoic. [1] Huiming Bao, J. Lyons, Chuanming Zhou (2008) Nature 453 504. [2] Huiming Bao, I.J. Fairchild, P.M. Wynn, C. Spötl (2009) Science 323 119. [3] B. Luz, E. Barkan, M. L. Bender, M. H. Thiemens, K. A. Boering (1999) Nature 400 547. [4] T. Blunier, B. Barnett, M. L. Bender, M. B. Hendricks (2002) Global Biogeochem. Cycles 16 doi:10.1029/2001GB001460. [5] D. L. Royer (2006) Geochim. Cosmochim. Acta 70 5665.

Understanding Microbial Contributions to Planetary Atmosphere

NASA Technical Reports Server (NTRS)

DesMarais, David J.

2000-01-01

Should our search of distant, extrasolar planetary atmospheres encounter evidence of life, that evidence will most likely be the gaseous products of microorganisms. Our biosphere was exclusively microbial for over 80 percent of its history and, even today, microbes strongly influence atmospheric composition. Life's greatest environmental impact arises from its capacity for harvesting energy and creating organic matter. Microorganisms catalyze the equilibration of C, S and transition metal species at temperatures where such reactions can be very slow in the absence of life. Sunlight has been harvested through photosynthesis to create enormous energy reservoirs that exist in the form of coexisting reservoirs of reduced, organic C and S stored in Earth's crust, and highly oxidized species (oxygen, sulfate and ferric iron) stored in the crust, oceans and atmosphere. Our civilization taps that storehouse of energy by burning fossil fuels. As astrobiologists, we identify the chemical consequences of distant biospheres as expressed in the atmospheres of their planets. Our approach must recognize that planets, biospheres and atmospheres evolve and change. For example, a tectonically more active early Earth hosted a thermophilic, non-photosynthetic biosphere and a mildly reducing, carbon dioxide-rich and oxygen-poor atmosphere. Microorganisms acquired energy by consuming hydrogen and sulfide and producing a broad array of reduced C and S gases, most notably, methane. Later, diverse types of bacterial photosynthesis developed that enhanced productivity but were incapable of splitting water to produce oxygen. Later, but still prior to 2.6 billion years ago, oxygenic photosynthesis developed. We can expect to encounter distant biospheres that represent various stages of evolution and that coexist with atmospheres ranging from mildly reducing to oxidizing compositions. Accordinaly, we must be prepared to interpret a broad range of atmospheric compositions, all containing signatures of life. Remarkably little is known about the composition of our own earlier atmosphere, particularly prior to the rise of oxygen levels some 2.0 to 2.2 billion years ago. Thus, field and laboratory observations and theoretical simulations should be conducted to examine the relationships between the structure and function of microbial ecosystems and their gaseous products. Ecosystems that are analogs of our ancient biosphere (e.g., based upon chemosynthesis or non-oxygenic photosynthesis, thermophilic and subsurface communities, etc.) should be included. Because key environmental parameters such as temperature and levels of hydrogen, carbon dioxide and oxygen varied during planetary evolution, their consequences for microbial ecosystems should be explored.

Ocean Drilling Program Contributions to the Understanding of the Deep Subsurface Biosphere

NASA Astrophysics Data System (ADS)

Fisk, M. R.

2003-12-01

Tantalizing evidence for microbes in oceanic basalts has been reported for a few decades, but it was from rocks cored on Ocean Drilling Program (ODP) Leg 148 in 1993 that the first clear-cut evidence of microbial invasion of ocean basalts was obtained. (Work on ODP legs, starting with Leg 112 in 1986, had already revealed the presence of significant microbial biomass in sediments.) In 1997 ODP created the Deep Biosphere Program Planning Group to promote the investigation of the microbiology of the ocean crust. In 1999 ODP built a microbiology lab on the JOIDES Resolution, and used the lab that year (Legs 185 and 187) to test the amount of microbial contamination introduced into rocks during drilling and to establish cultures from cored basalts. These experiments have been repeated on several legs since then. The development of CORKs has permitted long-term sampling of subseafloor fluids, and microorganisms have been recovered from CORKed holes. Thus, ODP made it possible for the scientific community to address major questions about the biology of the igneous crust, such as, (1) What microbes are present? (2) How abundant are they? (3) How are they distributed? DNA from basalts and subseafloor fluids reveal what types of organisms are present. Cell abundance and biomass have been estimated based on cell counts and on organic content of basalts. Surveys of basalts in DSDP/ODP repositories indicate that microorganisms are ubiquitous in the igneous crust. Microorganisms are found in rocks that are close to 100° C. They are found as deep as 1500 m below the sea floor, and in rocks as young as a few years and as old as 170 million years. Because of the vast size of the habitat, microorganism, even if present in small numbers, could be a significant fraction of the Earth's biomass. In a short time ODP contributed to advances in our understanding of the oceanic subsurface biosphere. Answers to other significant questions such as: (1) How do the microorganisms live?, (2) What impact do subsurface microorganisms have on the surface biosphere? (3) And, what roles do the subsurface biosphere play in element cycling? will be answered by future drilling. The International Ocean Drilling Program (IODP) is in the enviable position of providing support to address these key questions about the Earth's subsurface biosphere.

Modern Microbial Ecosystems are a Key to Understanding Our Biosphere's Early Evolution and its Contributions To The Atmosphere and Rock Record

NASA Technical Reports Server (NTRS)

DesMarais, David J.; DeVincenzi, Donald L. (Technical Monitor)

2000-01-01

The survival of our early biosphere depended upon efficient coordination anion- diverse microbial populations. Microbial mats exhibit a 3.46-billion-year fossil record, thus they are the oldest known ecosystems. Photosynthetic microbial mats were key because, today, sunlight powers more than 99 percent of global primary productivity. Thus photosynthetic ecosystems have affected the atmosphere profoundly and have created the most pervasive, easily-detected fossils. Photosynthetic biospheres elsewhere will be most detectible via telescopes or spacecraft. As a part of the Astrobiology Institute, our Ames Microbial Ecosystems group examines the roles played by ecological processes in the early evolution of our biosphere, as recorded in geologic fossils and in the macromolecules of living cells: (1) We are defining the microbial mat microenvironment, which was an important milieu for early evolution. (2) We are comparing mats in contrasting environments to discern strategies of adaptation and diversification, traits that were key for long-term survival. (3) We have selected sites that mimic key environmental attributes of early Earth and thereby focus upon evolutionary adaptations to long-term changes in the global environment. (4) Our studies of gas exchange contribute to better estimates of biogenic gases in Earth's early atmosphere. This group therefore directly addresses the question: How have the Earth and its biosphere influenced each other over time Our studies strengthen the systematics for interpreting the microbial fossil record and thereby enhance astrobiological studies of martian samples. Our models of biogenic gas emissions will enhance models of atmospheres that might be detected on inhabited extrasolar planets. This work therefore also addresses the question: How can other biospheres be recogniZed" Our choice of field sites helps us explore Earth's evolving early environment. For example, modern mats that occupy thermal springs and certain freshwater environments experience conditions such as low O2 and sulfate and high inorganic carbon and sulfide levels that resemble those of ancient marine environments. Later in history, both biologically-induced carbonate precipitation and the trapping and binding of suspended grains of carbonate became a dominant mechanism for carbonate deposition. Modern marine carbonate platforms and alkaline offer good examples of microbiologically-induced calcification. Both marine platforms and solar salterns illustrate microbially-driven trapping and binding. We are also exploring the effects of water composition upon the exchange of biogenic gases with the atmosphere.

Biodiversity in Oscypek, a Traditional Polish Cheese, Determined by Culture-Dependent and -Independent Approaches

PubMed Central

Alegría, Ángel; Szczesny, Pawel; Mayo, Baltasar; Bardowski, Jacek

2012-01-01

Oscypek is a traditional Polish scalded-smoked cheese, with a protected-designation-of-origin (PDO) status, manufactured from raw sheep's milk without starter cultures in the Tatra Mountains region of Poland. This study was undertaken in order to gain insight into the microbiota that develops and evolves during the manufacture and ripening stages of Oscypek. To this end, we made use of both culturing and the culture-independent methods of PCR followed by denaturing gradient gel electrophoresis (PCR-DGGE) and pyrosequencing of 16S rRNA gene amplicons. The culture-dependent technique and PCR-DGGE fingerprinting detected the predominant microorganisms in traditional Oscypek, whereas the next-generation sequencing technique (454 pyrosequencing) revealed greater bacterial diversity. Besides members of the most abundant bacterial genera in dairy products, e.g., Lactococcus, Lactobacillus, Leuconostoc, Streptococcus, and Enterococcus, identified by all three methods, other, subdominant bacteria belonging to the families Bifidobacteriaceae and Moraxellaceae (mostly Enhydrobacter), as well as various minor bacteria, were identified by pyrosequencing. The presence of bifidobacterial sequences in a cheese system is reported for the first time. In addition to bacteria, a great diversity of yeast species was demonstrated in Oscypek by the PCR-DGGE method. Culturing methods enabled the determination of a number of viable microorganisms from different microbial groups and their isolation for potential future applications in specific cheese starter cultures. PMID:22247135

Pyrosequencing analysis yields comprehensive assessment of microbial communities in pilot-scale two-stage membrane biofilm reactors.

PubMed

Ontiveros-Valencia, Aura; Tang, Youneng; Zhao, He-Ping; Friese, David; Overstreet, Ryan; Smith, Jennifer; Evans, Patrick; Rittmann, Bruce E; Krajmalnik-Brown, Rosa

2014-07-01

We studied the microbial community structure of pilot two-stage membrane biofilm reactors (MBfRs) designed to reduce nitrate (NO3(-)) and perchlorate (ClO4(-)) in contaminated groundwater. The groundwater also contained oxygen (O2) and sulfate (SO4(2-)), which became important electron sinks that affected the NO3(-) and ClO4(-) removal rates. Using pyrosequencing, we elucidated how important phylotypes of each "primary" microbial group, i.e., denitrifying bacteria (DB), perchlorate-reducing bacteria (PRB), and sulfate-reducing bacteria (SRB), responded to changes in electron-acceptor loading. UniFrac, principal coordinate analysis (PCoA), and diversity analyses documented that the microbial community of biofilms sampled when the MBfRs had a high acceptor loading were phylogenetically distant from and less diverse than the microbial community of biofilm samples with lower acceptor loadings. Diminished acceptor loading led to SO4(2-) reduction in the lag MBfR, which allowed Desulfovibrionales (an SRB) and Thiothrichales (sulfur-oxidizers) to thrive through S cycling. As a result of this cooperative relationship, they competed effectively with DB/PRB phylotypes such as Xanthomonadales and Rhodobacterales. Thus, pyrosequencing illustrated that while DB, PRB, and SRB responded predictably to changes in acceptor loading, a decrease in total acceptor loading led to important shifts within the "primary" groups, the onset of other members (e.g., Thiothrichales), and overall greater diversity.

Using pyrosequencing to shed light on deep mine microbial ecology

PubMed Central

Edwards, Robert A; Rodriguez-Brito, Beltran; Wegley, Linda; Haynes, Matthew; Breitbart, Mya; Peterson, Dean M; Saar, Martin O; Alexander, Scott; Alexander, E Calvin; Rohwer, Forest

2006-01-01

Background Contrasting biological, chemical and hydrogeological analyses highlights the fundamental processes that shape different environments. Generating and interpreting the biological sequence data was a costly and time-consuming process in defining an environment. Here we have used pyrosequencing, a rapid and relatively inexpensive sequencing technology, to generate environmental genome sequences from two sites in the Soudan Mine, Minnesota, USA. These sites were adjacent to each other, but differed significantly in chemistry and hydrogeology. Results Comparisons of the microbes and the subsystems identified in the two samples highlighted important differences in metabolic potential in each environment. The microbes were performing distinct biochemistry on the available substrates, and subsystems such as carbon utilization, iron acquisition mechanisms, nitrogen assimilation, and respiratory pathways separated the two communities. Although the correlation between much of the microbial metabolism occurring and the geochemical conditions from which the samples were isolated could be explained, the reason for the presence of many pathways in these environments remains to be determined. Despite being physically close, these two communities were markedly different from each other. In addition, the communities were also completely different from other microbial communities sequenced to date. Conclusion We anticipate that pyrosequencing will be widely used to sequence environmental samples because of the speed, cost, and technical advantages. Furthermore, subsystem comparisons rapidly identify the important metabolisms employed by the microbes in different environments. PMID:16549033

De Novo Transcriptome of the Hemimetabolous German Cockroach (Blattella germanica)

PubMed Central

Zhou, Xiaojie; Qian, Kun; Tong, Ying; Zhu, Junwei Jerry; Qiu, Xinghui; Zeng, Xiaopeng

2014-01-01

Background The German cockroach, Blattella germanica, is an important insect pest that transmits various pathogens mechanically and causes severe allergic diseases. This insect has long served as a model system for studies of insect biology, physiology and ecology. However, the lack of genome or transcriptome information heavily hinder our further understanding about the German cockroach in every aspect at a molecular level and on a genome-wide scale. To explore the transcriptome and identify unique sequences of interest, we subjected the B. germanica transcriptome to massively parallel pyrosequencing and generated the first reference transcriptome for B. germanica. Methodology/Principal Findings A total of 1,365,609 raw reads with an average length of 529 bp were generated via pyrosequencing the mixed cDNA library from different life stages of German cockroach including maturing oothecae, nymphs, adult females and males. The raw reads were de novo assembled to 48,800 contigs and 3,961 singletons with high-quality unique sequences. These sequences were annotated and classified functionally in terms of BLAST, GO and KEGG, and the genes putatively coding detoxification enzyme systems, insecticide targets, key components in systematic RNA interference, immunity and chemoreception pathways were identified. A total of 3,601 SSRs (Simple Sequence Repeats) loci were also predicted. Conclusions/Significance The whole transcriptome pyrosequencing data from this study provides a usable genetic resource for future identification of potential functional genes involved in various biological processes. PMID:25265537

Proteolytic bacterial dominance in a full-scale municipal solid waste anaerobic reactor assessed by 454 pyrosequencing technology.

PubMed

Cardinali-Rezende, Juliana; Rojas-Ojeda, Patricia; Nascimento, Andréa M A; Sanz, José L

2016-03-01

Biomethanization entails a good means to reduce the organic fraction (OF) derived from municipal solid wastes (MSW). The bacterial diversity of a full scale MSW anaerobic reactor located in Madrid (Spain) was investigated using high-throughput 454 pyrosequencing. Even though the proteolytic bacteria prevailed throughout all of the process, community shifts were observed from the start-up to the steady-state conditions, with an increasing biodiversity displayed over time. The Bacteroidetes and the Firmicutes were the majority phyla: 55.1 and 40.2% (start-up) and 18.7 and 78.7 (steady-state) of the total reads. The system's lack of evenness remains noteworthy as the sequences affiliated to the proteolytic non-saccharolytic Proteiniphylum, Gallicola and Fastidiosipila genera, together with the saccharolytic Saccharofermentans, were predominant on the system and this predominance appears to correlate with the presence of a high ammonium concentration. The 454 pyrosequencing revealed a great diversity of rare organisms which seemingly do not sustain any metabolic roles in the course of the OF-MSW degradation. However, this scarce and unique microbiota can confer great resilience to the system as a buffer against nutritional and environmental changing conditions, thus opening the door to increase the current knowledge about the bacterial community dynamics taking place during MSW treatment processes. Copyright © 2015 Elsevier Ltd. All rights reserved.

Random Amplification and Pyrosequencing for Identification of Novel Viral Genome Sequences

PubMed Central

Hang, Jun; Forshey, Brett M.; Kochel, Tadeusz J.; Li, Tao; Solórzano, Víctor Fiestas; Halsey, Eric S.; Kuschner, Robert A.

2012-01-01

ssRNA viruses have high levels of genomic divergence, which can lead to difficulty in genomic characterization of new viruses using traditional PCR amplification and sequencing methods. In this study, random reverse transcription, anchored random PCR amplification, and high-throughput pyrosequencing were used to identify orthobunyavirus sequences from total RNA extracted from viral cultures of acute febrile illness specimens. Draft genome sequence for the orthobunyavirus L segment was assembled and sequentially extended using de novo assembly contigs from pyrosequencing reads and orthobunyavirus sequences in GenBank as guidance. Accuracy and continuous coverage were achieved by mapping all reads to the L segment draft sequence. Subsequently, RT-PCR and Sanger sequencing were used to complete the genome sequence. The complete L segment was found to be 6936 bases in length, encoding a 2248-aa putative RNA polymerase. The identified L segment was distinct from previously published South American orthobunyaviruses, sharing 63% and 54% identity at the nucleotide and amino acid level, respectively, with the complete Oropouche virus L segment and 73% and 81% identity at the nucleotide and amino acid level, respectively, with a partial Caraparu virus L segment. The result demonstrated the effectiveness of a sequence-independent amplification and next-generation sequencing approach for obtaining complete viral genomes from total nucleic acid extracts and its use in pathogen discovery. PMID:22468136

Barcoded pyrosequencing analysis of the microbial community in a simulator of the human gastrointestinal tract showed a colon region-specific microbiota modulation for two plant-derived polysaccharide blends.

PubMed

Marzorati, Massimo; Maignien, Lois; Verhelst, An; Luta, Gabriela; Sinnott, Robert; Kerckhof, Frederiek Maarten; Boon, Nico; Van de Wiele, Tom; Possemiers, Sam

2013-02-01

The combination of a Simulator of the Human Intestinal Microbial Ecosystem with ad hoc molecular techniques (i.e. pyrosequencing, denaturing gradient gel electrophoresis and quantitative PCR) allowed an evaluation of the extent to which two plant polysaccharide supplements could modify a complex gut microbial community. The presence of Aloe vera gel powder and algae extract in product B as compared to the standard blend (product A) improved its fermentation along the entire simulated colon. The potential extended effect of product B in the simulated distal colon, as compared to product A, was confirmed by: (i) the separate clustering of the samples before and after the treatment in the phylogenetic-based dendrogram and OTU-based PCoA plot only for product B; (ii) a higher richness estimator (+33 vs. -36 % of product A); and (iii) a higher dynamic parameter (21 vs. 13 %). These data show that the combination of well designed in vitro simulators with barcoded pyrosequencing is a powerful tool for characterizing changes occurring in the gut microbiota following a treatment. However, for the quantification of low-abundance species-of interest because of their relationship to potential positive health effects (i.e. bifidobacteria or lactobacilli)-conventional molecular ecological approaches, such as PCR-DGGE and qPCR, still remain a very useful complementary tool.

Gene discovery using next-generation pyrosequencing to develop ESTs for Phalaenopsis orchids

PubMed Central

2011-01-01

Background Orchids are one of the most diversified angiosperms, but few genomic resources are available for these non-model plants. In addition to the ecological significance, Phalaenopsis has been considered as an economically important floriculture industry worldwide. We aimed to use massively parallel 454 pyrosequencing for a global characterization of the Phalaenopsis transcriptome. Results To maximize sequence diversity, we pooled RNA from 10 samples of different tissues, various developmental stages, and biotic- or abiotic-stressed plants. We obtained 206,960 expressed sequence tags (ESTs) with an average read length of 228 bp. These reads were assembled into 8,233 contigs and 34,630 singletons. The unigenes were searched against the NCBI non-redundant (NR) protein database. Based on sequence similarity with known proteins, these analyses identified 22,234 different genes (E-value cutoff, e-7). Assembled sequences were annotated with Gene Ontology, Gene Family and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Among these annotations, over 780 unigenes encoding putative transcription factors were identified. Conclusion Pyrosequencing was effective in identifying a large set of unigenes from Phalaenopsis. The informative EST dataset we developed constitutes a much-needed resource for discovery of genes involved in various biological processes in Phalaenopsis and other orchid species. These transcribed sequences will narrow the gap between study of model organisms with many genomic resources and species that are important for ecological and evolutionary studies. PMID:21749684

Assessment of replicate bias in 454 pyrosequencing and a multi-purpose read-filtering tool.

PubMed

Jérôme, Mariette; Noirot, Céline; Klopp, Christophe

2011-05-26

Roche 454 pyrosequencing platform is often considered the most versatile of the Next Generation Sequencing technology platforms, permitting the sequencing of large genomes, the analysis of variations or the study of transcriptomes. A recent reported bias leads to the production of multiple reads for a unique DNA fragment in a random manner within a run. This bias has a direct impact on the quality of the measurement of the representation of the fragments using the reads. Other cleaning steps are usually performed on the reads before assembly or alignment. PyroCleaner is a software module intended to clean 454 pyrosequencing reads in order to ease the assembly process. This program is a free software and is distributed under the terms of the GNU General Public License as published by the Free Software Foundation. It implements several filters using criteria such as read duplication, length, complexity, base-pair quality and number of undetermined bases. It also permits to clean flowgram files (.sff) of paired-end sequences generating on one hand validated paired-ends file and the other hand single read file. Read cleaning has always been an important step in sequence analysis. The pyrocleaner python module is a Swiss knife dedicated to 454 reads cleaning. It includes commonly used filters as well as specialised ones such as duplicated read removal and paired-end read verification.

USSR and Eastern Europe Scientific Abstracts. Geophysics, Astronomy and Space, Number 430

DTIC Science & Technology

1978-10-02

making the necessary preparations. (PRAVDA 30 Aug 78 p 1) 30 AUGUST 31 AUGUST In accordance with the " Biosfera " (biosphere) experiment, the...On their sixth day on board the space complex, cosmonauts Bykovskiy and Jaehn continued the " Biosfera " environmental studies experiment as well as...AUGUST In accordance with the " Biosfera " (biosphere) experiment, the internatxonal crew conducted observations of the earth’s surface, a variety of

Our Changing Planet: The U.S. Climate Change Science Program for Fiscal Year 2009

DTIC Science & Technology

2008-07-01

forests of the southern Yucatan Peninsula form the largest expanse of this ecosystem type remaining in Mexico. It forms an ecocline between a drier...Calakmul Biosphere Reserve to preserve this unique forested area. The Southern Yucatan Peninsular Region Project is currently engaged in an assessment of the...Eastman, B.L. Turner II, S. Calme, R. Dickson, C. Pozo, and F. Sangermano, 2007: Land Change in the southern Yucatan and Calamul biosphere reserve

Multiple greenhouse-gas feedbacks from the land biosphere under future climate change scenarios

NASA Astrophysics Data System (ADS)

Stocker, Benjamin D.; Roth, Raphael; Joos, Fortunat; Spahni, Renato; Steinacher, Marco; Zaehle, Soenke; Bouwman, Lex; Xu-Ri; Prentice, Iain Colin

2013-07-01

Atmospheric concentrations of the three important greenhouse gases (GHGs) CO2, CH4 and N2O are mediated by processes in the terrestrial biosphere that are sensitive to climate and CO2. This leads to feedbacks between climate and land and has contributed to the sharp rise in atmospheric GHG concentrations since pre-industrial times. Here, we apply a process-based model to reproduce the historical atmospheric N2O and CH4 budgets within their uncertainties and apply future scenarios for climate, land-use change and reactive nitrogen (Nr) inputs to investigate future GHG emissions and their feedbacks with climate in a consistent and comprehensive framework. Results suggest that in a business-as-usual scenario, terrestrial N2O and CH4 emissions increase by 80 and 45%, respectively, and the land becomes a net source of C by AD 2100. N2O and CH4 feedbacks imply an additional warming of 0.4-0.5°C by AD 2300; on top of 0.8-1.0°C caused by terrestrial carbon cycle and Albedo feedbacks. The land biosphere represents an increasingly positive feedback to anthropogenic climate change and amplifies equilibrium climate sensitivity by 22-27%. Strong mitigation limits the increase of terrestrial GHG emissions and prevents the land biosphere from acting as an increasingly strong amplifier to anthropogenic climate change.

[The high pressure life of piezophiles].

PubMed

Oger, Philippe; Cario, Anaïs

2014-01-01

The deep biosphere is composed of very different biotopes located in the depth of the oceans, the ocean crust or the lithosphere. Although very different, deep biosphere biotopes share one common feature, high hydrostatic pressure. The deep biosphere is colonized by specific organisms, called piezophiles, that are able to grow under high hydrostatic pressure. Bacterial piezophiles are mainly psychrophiles belonging to five genera of γ-proteobacteria, Photobacterium, Shewanella, Colwellia, Psychromonas and Moritella, while piezophilic Archaea are mostly (hyper)thermophiles from the Thermococcales. None of these genera are specific for the deep biosphere. High pressure deeply impacts the activity of cells and cellular components, and reduces the activity of numerous key processes, eventually leading to cell death of piezosensitive organisms. Biochemical and genomic studies yield a fragmented view on the adaptive mechanisms in piezophiles. It is yet unclear whether piezophilic adaptation requires the modification of a few genes, or metabolic pathways, or a more profound reorganization of the genome, the fine tuning of gene expression to compensate the pressure-induced loss of activity of the proteins most affected by high pressure, or a stress-like physiological cell response. In contrast to what has been seen for thermophily or halophily, the adaptation to high pressure is diffuse in the genome and may concern only a small fraction of the genes. © Société de Biologie, 2014.

Constraining Centennial-Scale Ecosystem-Climate Interactions with a Pre-colonial Forest Reconstruction across the Upper Midwest and Northeastern United States

NASA Astrophysics Data System (ADS)

Matthes, J. H.; Dietze, M.; Fox, A. M.; Goring, S. J.; McLachlan, J. S.; Moore, D. J.; Poulter, B.; Quaife, T. L.; Schaefer, K. M.; Steinkamp, J.; Williams, J. W.

2014-12-01

Interactions between ecological systems and the atmosphere are the result of dynamic processes with system memories that persist from seconds to centuries. Adequately capturing long-term biosphere-atmosphere exchange within earth system models (ESMs) requires an accurate representation of changes in plant functional types (PFTs) through time and space, particularly at timescales associated with ecological succession. However, most model parameterization and development has occurred using datasets than span less than a decade. We tested the ability of ESMs to capture the ecological dynamics observed in paleoecological and historical data spanning the last millennium. Focusing on an area from the Upper Midwest to New England, we examined differences in the magnitude and spatial pattern of PFT distributions and ecotones between historic datasets and the CMIP5 inter-comparison project's large-scale ESMs. We then conducted a 1000-year model inter-comparison using six state-of-the-art biosphere models at sites that bridged regional temperature and precipitation gradients. The distribution of ecosystem characteristics in modeled climate space reveals widely disparate relationships between modeled climate and vegetation that led to large differences in long-term biosphere-atmosphere fluxes for this region. Model simulations revealed that both the interaction between climate and vegetation and the representation of ecosystem dynamics within models were important controls on biosphere-atmosphere exchange.

Impact craters as biospheric microenvironments, Lawn Hill Structure, Northern Australia.

PubMed

Lindsay, John; Brasier, Martin

2006-04-01

Impact craters on Mars act as traps for eolian sediment and in the past may have provided suitable microenvironments that could have supported and preserved a stressed biosphere. If this is so, terrestrial impact structures such as the 18-km-diameter Lawn Hill Structure, in northern Australia, may prove useful as martian analogs. We sampled outcrop and drill core from the carbonate fill of the Lawn Hill Structure and recorded its gamma-log signature. Facies data along with whole rock geochemistry and stable isotope signatures show that the crater fill is an outlier of the Georgina Basin and was formed by impact at, or shortly before, approximately 509-506 million years ago. Subsequently, it was rapidly engulfed by the Middle Cambrian marine transgression, which filled it with shallow marine carbonates and evaporites. The crater formed a protected but restricted microenvironment in which sediments four times the thickness of the nearby basinal succession accumulated. Similar structures, common on the martian surface, may well have acted as biospheric refuges as the planet's water resources declined. Low-pH aqueous environments on Earth similar to those on Mars, while extreme, support diverse ecologies. The architecture of the eolian crater fill would have been defined by long-term ground water cycles resulting from intermittent precipitation in an extremely arid climate. Nutrient recycling, critical to a closed lacustrine sub-ice biosphere, could be provided by eolian transport onto the frozen water surface.

Predicting what extra-terrestrials will be like: and preparing for the worst.

PubMed

Morris, Simon Conway

2011-02-13

It is difficult to imagine evolution in alien biospheres operating in any manner other than Darwinian. Yet, it is also widely assumed that alien life-forms will be just that: strange, un-nerving and probably repulsive. There are two reasons for this view. First, it is assumed that the range of habitable environments available to extra-terrestrial life is far wider than on Earth. I suggest, however, that terrestrial life is close to the physical and chemical limits of life anywhere. Second, it is a neo-Darwinian orthodoxy that evolution lacks predictability; imagining what extra-terrestrial life would look like in any detail is a futile exercise. To the contrary, I suggest that the outcomes of evolution are remarkably predictable. This, however, leads us to consider two opposites, both of which should make our blood run cold. The first, and actually extremely unlikely, is that alien biospheres will be strikingly similar to our terrestrial equivalent and that in such biospheres intelligence will inevitably emerge. The reasons for this revolve around the ubiquity of evolutionary convergence, the determinate structure of the Tree of Life and molecular inherency. But if something like a human is an inevitability, why do I also claim that the first possibility is 'extremely unlikely'? Simply because the other possibility is actually the correct answer. Paradoxically, we and our biosphere are completely alone. So which is worse? Meeting ourselves or meeting nobody?

Developing Starlight connections with UNESCO sites through the Biosphere Smart

NASA Astrophysics Data System (ADS)

Marin, Cipriano

2015-08-01

The large number of UNESCO Sites around the world, in outstanding sites ranging from small islands to cities, makes it possible to build and share a comprehensive knowledge base on good practices and policies on the preservation of the night skies consistent with the protection of the associated scientific, natural and cultural values. In this context, the Starlight Initiative and other organizations such as IDA play a catalytic role in an essential international process to promote comprehensive, holistic approaches on dark sky preservation, astronomical observation, environmental protection, responsible lighting, sustainable energy, climate change and global sustainability.Many of these places have the potential to become models of excellence to foster the recovery of the dark skies and its defence against light pollution, included some case studies mentioned in the Portal to the Heritage of Astronomy.Fighting light pollution and recovering starry sky are already elements of a new emerging culture in biosphere reserves and world heritage sites committed to acting on climate change and sustainable development. Over thirty territories, including biosphere reserves and world heritage sites, have been developed successful initiatives to ensure night sky quality and promote sustainable lighting. Clear night skies also provide sustainable income opportunities as tourists and visitors are eagerly looking for sites with impressive night skies.Taking into account the high visibility and the ability of UNESCO sites to replicate network experiences, the Starlight Initiative has launched an action In cooperation with Biosphere Smart, aimed at promoting the Benchmark sites.Biosphere Smart is a global observatory created in partnership with UNESCO MaB Programme to share good practices, and experiences among UNESCO sites. The Benchmark sites window allows access to all the information of the most relevant astronomical heritage sites, dark sky protected areas and other places committed to the preservation of the values associated with the night sky. A new step ahead in our common task of protecting the starry skies at UNESCO sites.

Evolvability Is an Evolved Ability: The Coding Concept as the Arch-Unit of Natural Selection.

PubMed

Janković, Srdja; Ćirković, Milan M

2016-03-01

Physical processes that characterize living matter are qualitatively distinct in that they involve encoding and transfer of specific types of information. Such information plays an active part in the control of events that are ultimately linked to the capacity of the system to persist and multiply. This algorithmicity of life is a key prerequisite for its Darwinian evolution, driven by natural selection acting upon stochastically arising variations of the encoded information. The concept of evolvability attempts to define the total capacity of a system to evolve new encoded traits under appropriate conditions, i.e., the accessible section of total morphological space. Since this is dependent on previously evolved regulatory networks that govern information flow in the system, evolvability itself may be regarded as an evolved ability. The way information is physically written, read and modified in living cells (the "coding concept") has not changed substantially during the whole history of the Earth's biosphere. This biosphere, be it alone or one of many, is, accordingly, itself a product of natural selection, since the overall evolvability conferred by its coding concept (nucleic acids as information carriers with the "rulebook of meanings" provided by codons, as well as all the subsystems that regulate various conditional information-reading modes) certainly played a key role in enabling this biosphere to survive up to the present, through alterations of planetary conditions, including at least five catastrophic events linked to major mass extinctions. We submit that, whatever the actual prebiotic physical and chemical processes may have been on our home planet, or may, in principle, occur at some time and place in the Universe, a particular coding concept, with its respective potential to give rise to a biosphere, or class of biospheres, of a certain evolvability, may itself be regarded as a unit (indeed the arch-unit) of natural selection.

Multilevel approach to mentoring in the Research Experiences for Undergraduates programs

NASA Astrophysics Data System (ADS)

Bonine, K. E.; Dontsova, K.; Pavao-Zuckerman, M.; Paavo, B.; Hogan, D.; Oberg, E.; Gay, J.

2015-12-01

This presentation focuses on different types of mentoring for students participating in Research Experiences for Undergraduates programs with examples, including some new approaches, from The Environmental and Earth Systems Research Experiences for Undergraduates Program at Biosphere 2. While traditional faculty mentors play essential role in students' development as researchers and professionals, other formal and informal mentoring can be important component of the REU program and student experiences. Students receive mentoring from program directors, coordinators, and on site undergraduate advisors. While working on their research projects, REU students receive essential support and mentoring from undergraduate and graduate students and postdoctoral scientists in the research groups of their primary mentors. Cohort living and group activities give multiple opportunities for peer mentoring where each student brings their own strengths and experiences to the group. Biosphere 2 REU program puts strong emphasis on teaching students to effectively communicate their research to public. In order to help REUs learn needed skills the outreach personnel at Biosphere 2 mentor and advise students both in groups and individually, in lecture format and by personal example, on best outreach approaches in general and on individual outreach projects students develop. To further enhance and strengthen outreach mentoring we used a novel approach of blending cohort of REU students with the Cal Poly STAR (STEM Teacher And Researcher) Program fellows, future K-12 STEM teachers who are gaining research experience at Biosphere 2. STAR fellows live together with the REU students and participate with them in professional development activities, as well as perform research side by side. Educational background and experiences gives these students a different view and better preparation and tools to effectively communicate and adapt science to lay audiences, a challenge commonly facing researchers but rarely taught to future scientists. In addition, REU students act as mentors themselves to the middle and high school students in Biosphere 2 Science Academy sharing with them exciting research they are doing and their experiences about doing science and life in college.

What are the Outstanding Questions in Biosphere-Atmosphere Exchange?

NASA Astrophysics Data System (ADS)

Katul, G.; Finnigan, J.

2002-12-01

Our answer to this question has three parts. At the highest level human population is now so large (and will grow to 10-12 million by 2050 before stabilizing and reversing) and our capacity to influence the biosphere is so profound that we must now treat human, biophysical and geophysical processes as a single interacting system: the Anthroposphere. The Anthroposphere is a complex adaptive system (CAS) in the sense that action of humans individually or in groups (as large as nations) are a response to an environment that includes not just the geosphere and biosphere but the sum of human interactions as well. These actions, in turn, will change this system that provides critical ecosystem services that we are just now realizing are finite and threatened. At present we lack the tools and understanding even to frame key questions about the Anthroposphere although the emerging discipline of Complex System Science is hinting at ways this might be done. At the next level we address the problems posed by cross-scale interactions central to the state and function of the geosphere and biosphere, where processes at the cellular level, (e.g. photosynthesis and respiration), have consequences on the global level and vice-versa through a sequence of non-linear upscale and downscale interactions. Currently there are large gaps in our understanding of this scale cascade that inhibit our ability to parameterize, model and predict phenomena such as the rapid shifts in climate that recent studies of the paleo record have revealed. In short, we need to quantify the risk that our current actions may cause abrupt climate changes that may then feed upon the Anthrosphere. Unless we can reduce the uncertainty with which we describe quantitatively the non-human parts of the Anthroposphere, the injection of models of human interaction, initially tentative, may reduce our predictive ability to unusable levels. Finally we can detail a set of key processes that are themselves critical to geosphere-biosphere functioning but which are poorly understood as yet.

Top-Down Assessment of the Asian Carbon Budget Since the Mid 1990s

NASA Astrophysics Data System (ADS)

Thompson, R.; Canadell, J.; Patra, P. K.; Chevallier, F.; Maksyutov, S. S.; Law, R. M.; Ziehn, T.; van der Laan-Luijkx, I. T.; Peters, W.; Ganshin, A.; Zhuravlev, R.; Maki, T.; Nakamura, T.; Shirai, T.; Ishizawa, M.; Saeki, T.; Poulter, B.; Ciais, P.

2015-12-01

Atmospheric carbon dioxide (CO2) is the principle driver of anthropogenic climate change. Asia is an important region for the global carbon budget, with four of the world's ten largest national emitters of CO2, but it is also a region with considerable uncertainty in both anthropogenic emissions and land biosphere fluxes of CO2. Furthermore, Asia has undergone rapid economic growth over the past two decades, which has been associated with large increases in fossil fuel emissions, 190% for India and 240% for China between 1990 and 2010. We have used an ensemble of seven atmospheric CO2 inversions and three standard fossil fuel and cement fluxes, based on the inventories of CDIAC, EDGAR and IEA, to determine the land biosphere fluxes for East, South and Southeast Asia, and to ascertain the robustness and overall uncertainty of the results. We find that the East Asian land biosphere was on average a carbon sink of -0.35 ± 0.37 PgC y-1 (median and MAD), or equivalently 17 ± 18% of East Asia's fossil fuel and cement emissions, over 1996 - 2012. Between 1996 - 2001 and 2008 - 2012, we find an increase in the sink of 0.74 ± 0.28 PgC y-1, however the magnitude of this is contingent on the assumed increase in fossil fuel emissions. For South Asia, we find that on average the land biosphere was close to carbon neutral, -0.01 ± 0.20 PgC y-1 over 1996 - 2012 and that there was no significant trend. For Southeast Asia, we find no evidence for a trend in the land biosphere flux over 1996 - 2012 and we cannot determine any difference from carbon neutrality (as assumed a priori by most inversions) with a flux of 0.06 ± 0.29 PgC y-1, throughout this period despite extensive tropical deforestation.

Benthic δ13C stacks: Metrics for deglacial changes in deep ocean carbon storage and the terrestrial biosphere

NASA Astrophysics Data System (ADS)

Peterson, C.; Lisiecki, L. E.

2016-12-01

Across the deglaciation, atmospheric CO2 and global temperatures rise while the deep ocean ventilates carbon to the atmosphere and terrestrial biosphere. As the terrestrial biosphere expands, the mean global ocean δ13C signature increases in response. How well constrained is the global mean benthic δ13C from 20-6 ka? Are the atmosphere and terrestrial biosphere signals in benthic δ13C coupled across the deglaciation? Improved understanding of deglacial carbon cycle interactions can help close the gap between data-based and model-based estimates of global mean benthic δ13C and deep ocean carbon storage changes. Here we present a 118-record compilation of Cibicides wuellerstorfi δ13C time series that span 20-6 kyr. The δ13C records with a resolution better than 3 kyr and gaps between data smaller than 4 kyr are aligned to age models that are constrained by planktic 14C ages (Stern and Lisiecki, 2014). The δ13C records are stacked within nine regions. Then these regional stacks are combined using volume-weighted averages to create intermediate, deep and whole ocean δ13C stacks. The δ13C gradient between the intermediate and deep stacks covaries with atmospheric CO2 change. Meanwhile the deglacial global ocean mean δ13C rise tracks the expansion of the global terrestrial biosphere from 19-6 ka. From this volume-weighted global δ13C stack, the LGM-Holocene mean δ13C change is 0.35±0.10‰ similar to previous estimates (Curry et al., 1988; Duplessy et al., 1988; Peterson et al., 2015; Gebbie et al., 2015). The δ13C stacks and this 4D δ13C compilation are ideal for model-data comparisons and time-stepping 3D visualizations.

Initial experimental results from the Laboratory Biosphere closed ecological system facility

NASA Astrophysics Data System (ADS)

Alling, A.; Allen, J.; Dempster, W.; Nelson, M.; Silverstone, S.; van Thillo, M.

Results from the closure and initial closed ecological system research in the "Laboratory Biosphere" facility in Santa Fe, New Mexico (USA) will be presented. The facility was initially sealed in April 2002; and the first crop experiments with soybeans commenced in May 2002. The Laboratory Biosphere was created by the team which invented, built and operated Biosphere 2 during its years of closed ecological system functioning (1991-94) and is a testbed to build upon the lessons learned. It is an opportunity to continue experiments with a sustainable soil based agriculture system unlike most bioregenerative systems which use hydroponic systems dependent on a supply of nutrient solution. Because of the small volume of the system (34-45 m3), developing mechanisms to keep parameters like carbon dioxide within acceptable limits will be critical. Recycle of nutrients within the system to maintain soil fertility; and the ability of the inherent complex ecology of soils and a soil bed reactor to handle trace gas buildups are primary research goals. Other research goals are determination of short and long-term exchanges between soil, plants and atmosphere, especially for carbon dioxide, oxygen, nitrogen, NOX, and methane, impact of cultivation (tillage) on soil/atmospheric exchanges., investigation and development of strategies to return nutrients to the soil to maintain fertility, e.g. shredding biomass vs. composting, impact on soil chemistry of returning leachate water to the soil as irrigation water. The microbiological status of soils prior to experiments and over time will allow measurement of changes in microbial diversity and the determination of the role of soil microbes in biogeochemical cycles. Integration of automated sensor and control in the system with real-time modeling has importance for operation, research and educational outreach programs. The Laboratory Biosphere is intended to test and develop a "cybersphere" (network of shared intelligence) that may be scaled up for natural ecosystems and the global environment.

Technical Review of the Laboratory Biosphere Closed Ecological System Facility

NASA Astrophysics Data System (ADS)

Dempster, W.; van Thillo, M.; Alling, A.; Allen, J.; Silverstone, S.; Nelson, M.

The "Laboratory Biosphere", a new closed ecological system facility in Santa Fe, New Mexico (USA) has been constructed and became operational in May 2002. Built and operated by the Global Ecotechnics consortium (Biosphere Technologies and Biosphere Foundation with Biospheric Design Inc., and the Institute of Ecotechnics), the research apparatus for intensive crop growth, biogeochemical cycle dynamics and recycling of inedible crop biomass comprises a sealed cylindrical steel chamber and attached variable volume chamber (lung) to prevent pressures caused by the expansion and contraction of the contained air. The cylindrical growing chamber is 3.7m (12 feet) long and 3.7m (12 foot) diameter, giving an internal volume of 34 m3 (1200 ft 3 ). The two crop growth beds cover 5.5 m2, with a soil depth of 0.3m (12 inches), with 12 x 1000 watt high-pressure sodium lights capable of variable lighting of 40-70 mol per m2 per day. A small soil bed reactor in the chamber can be activated to help with metabolism of chamber trace gases. The volume of the attached variable volume chamber (lung) can range between 0-11 m3 (0-400 ft 3 ). Evapotranspired and soil leachate water are collected, combined and recycled to water the planting beds. Sampling ports enable testing of water quality of leachate, condensate and irrigation water. Visual inspection windows provide views of the entire interior and growing beds. The chamber is also outfitted with an airlock to minimize air exchange when people enter and work in the chamber. Continuous sensors include atmospheric CO2 and oxygen, temperature, humidity, soil moisture, light level and water levels in reservoirs. Both "sniffer" (air ports) and "sipper" (water ports) will enable collection of water or air samples for detailed analysis. This paper reports on the development of this new soil-based bioregenerative life support closed system apparatus and its technical challenges and capabilities.

Assessment of Anthropogenic and Climatic Impacts on the Global Carbon Cycle Using a 3-D Model Constrained by Isotopic Carbon Measurements and Remote Sensing of Vegetation

NASA Technical Reports Server (NTRS)

Keeling, Charles D.; Piper, S. C.

1998-01-01

Our original proposal called for improved modeling of the terrestrial biospheric carbon cycle, specifically using biome-specific process models to account for both the energy and water budgets of plant growth, to facilitate investigations into recent changes in global atmospheric CO2 abundance and regional distribution. The carbon fluxes predicted by these models were to be incorporated into a global model of CO2 transport to establish large-scale regional fluxes of CO2 to and from the terrestrial biosphere subject to constraints imposed by direct measurements of atmospheric CO2 and its 13C/12C isotopic ratio. Our work was coordinated with a NASA project (NASA NAGW-3151) at the University of Montana under the direction of Steven Running, and was partially funded by the Electric Power Research Institute. The primary objective of this project was to develop and test the Biome-BGC model, a global biological process model with a daily time step which simulates the water, energy and carbon budgets of plant growth. The primary product, the unique global gridded daily land temperature, and the precipitation data set which was used to drive the process model is described. The Biome-BGC model was tested by comparison with a simpler biological model driven by satellite-derived (NDVI) Normalized Difference Vegetation Index and (PAR) Photosynthetically Active Radiation data and by comparison with atmospheric CO2 observations. The simple NDVI model is also described. To facilitate the comparison with atmospheric CO2 observations, a three-dimensional atmospheric transport model was used to produce predictions of atmospheric CO2 variations given CO2 fluxes owing to (NPP) Net Primary Productivity and heterotrophic respiration that were produced by the Biome-BGC model and by the NDVI model. The transport model that we used in this project, and errors associated with transport simulations, were characterized by a comparison of 12 transport models.

Preliminary characterization of the oral microbiota of Chinese adults with and without gingivitis

PubMed Central

2011-01-01

Background Microbial communities inhabiting human mouth are associated with oral health and disease. Previous studies have indicated the general prevalence of adult gingivitis in China to be high. The aim of this study was to characterize in depth the oral microbiota of Chinese adults with or without gingivitis, by defining the microbial phylogenetic diversity and community-structure using highly paralleled pyrosequencing. Methods Six non-smoking Chinese, three with and three without gingivitis (age range 21-39 years, 4 females and 2 males) were enrolled in the present cross-sectional study. Gingival parameters of inflammation and bleeding on probing were characterized by a clinician using the Mazza Gingival Index (MGI). Plaque (sampled separately from four different oral sites) and salivary samples were obtained from each subject. Sequences and relative abundance of the bacterial 16 S rDNA PCR-amplicons were determined via pyrosequencing that produced 400 bp-long reads. The sequence data were analyzed via a computational pipeline customized for human oral microbiome analyses. Furthermore, the relative abundances of selected microbial groups were validated using quantitative PCR. Results The oral microbiomes from gingivitis and healthy subjects could be distinguished based on the distinct community structures of plaque microbiomes, but not the salivary microbiomes. Contributions of community members to community structure divergence were statistically accessed at the phylum, genus and species-like levels. Eight predominant taxa were found associated with gingivitis: TM7, Leptotrichia, Selenomonas, Streptococcus, Veillonella, Prevotella, Lautropia, and Haemophilus. Furthermore, 98 species-level OTUs were identified to be gingivitis-associated, which provided microbial features of gingivitis at a species resolution. Finally, for the two selected genera Streptococcus and Fusobacterium, Real-Time PCR based quantification of relative bacterial abundance validated the pyrosequencing-based results. Conclusions This methods study suggests that oral samples from this patient population of gingivitis can be characterized via plaque microbiome by pyrosequencing the 16 S rDNA genes. Further studies that characterize serial samples from subjects (longitudinal study design) with a larger population size may provide insight into the temporal and ecological features of oral microbial communities in clinically-defined states of gingivitis. PMID:22152152

A brief description of the simple biosphere model (SiB)

NASA Technical Reports Server (NTRS)

Sellers, P. J.; Mintz, Y.; Sud, Y. C.

1986-01-01

A biosphere model for calculating the transfer of energy, mass, and momentum between the atmosphere and the vegetated surface of the Earth was designed for atmospheric general circulation models. An upper vegetation layer represents the perennial canopy of trees or shrubs, a lower layer represents the annual ground cover of grasses and other herbacious species. The local coverage of each vegetation layer may be fractional or complete but as the individual vegetation elements are considered to be evenly spaced, their root systems are assumed to extend uniformly throughout the entire grid-area. The biosphere has seven prognostic physical-state variables: two temperatures (one for the canopy and one for the ground cover and soil surface); two interception water stores (one for the canopy and one for the ground cover); and three soil moisture stores (two of which can be reached by the vegetation root systems and one underlying recharge layer into and out of which moisture is transferred only by hydraulic diffusion).

Mars Analog Rio Tinto Experiment (MARTE): 2003 Drilling Campaign to Search for a Subsurface Biosphere at Rio Tinto Spain

NASA Technical Reports Server (NTRS)

Stoker, Carol; Dunagan, Stephen; Stevens, Todd; Amils, Ricardo; Gomez-Elvira, Javier; Fernandez, David; Hall, James; Lynch, Kennda; Cannon, Howard; Zavaleta, Jhony

2004-01-01

The MARTE (Mars Astrobiology Research and Technology Experiment) project, an ASTEP field experiment, is exploring for a hypothesized subsurface anaerobic chemoautotrophic biosphere in the region of the Tinto River- or Rio Tinto- in southwestern Spain. It is also demonstrating technology needed to search for a subsurface biosphere on Mars. The project has three primary objectives: (1) search for and characterize subsurface life at Rio Tinto along with the physical and chemical properties and sustaining energy sources of its environment, (2) perform a high fidelity simulation of a robotic Mars drilling mission to search for life, and (3) demonstrate the drilling, sample handling, and instrument technologies relevant to searching for life on Mars. The simulation of the robotic drilling mission is guided by the results of the aseptic drilling campaign to search for life at Rio Tinto. This paper describes results of the first phase of the aseptic drilling campaign.

Cosmic Rays as a Factor of Biosphere Evolution

NASA Astrophysics Data System (ADS)

Miroshnichenko, L. I.

2014-11-01

There are no doubts that the Earth's space environment in the past inevitably exerted direct and/or indirect influence [1--4] on the conditions of terrestrial life and biospheric evolution. Well-known cosmic factors are usually streams of cosmic dust and gas, comets and asteroids, cosmic rays (energetic particles of galactic and/or solar origin), interplanetary plasma (solar wind), and electromagnetic radiation of different energies, wavelengths, or frequencies. Of great interest are radiation conditions and their variations, especially in the remote past (over the geological time scales). The Sun, the most important and indispensable condition for the existence of the Earth's biosphere, is also a potential source of dangerous emissions. In continuation of (and in addition to) our review paper [3], below we summarize the observational data and results of theoretical works that have been carried out and/or published mainly after 2012. These studies are actually in the frontier region between the Astrobiology and Space Weather. Our main attention is paid to cosmic rays (CR) of galactic and solar origin (GCR and SCR, respectively).

Agricultural ecosystems - The world is watching

DOE Office of Scientific and Technical Information (OSTI.GOV)

Madison, M.F.; Licht, L.A.

1990-02-01

Environmental degradation is displacing nuclear war as the overriding concern of the world's people. An accusing finger is rightfully pointed at agricultural practices - for degrading water, air, food, and societal quality. As reported in the popular and technical press, there is a clamor for farming technology that is both productive and ecological. We cannot survive without a productive agriculture. Yet, the eroding soil, the degrading water quality, the decrease in farm profitability, the reductions in wildlife populations, and the closing store fronts in rural America point to a need for new management approaches. The word sustainable continues to bemore » mentioned as an underlying theme for future management techniques. Soil, air, and water form a seamless whole - the thin envelope we call the biosphere. The term sustainable agriculture implies a nourishing stewardship of the biosphere when used by farmers in pursuit of their livelihood. This biosphere simultaneously produces and sustains a multitude of products, including ourselves. It is all we have to create both our present and our future.« less

Exploration and protection of Europa's biosphere: implications of permeable ice.

PubMed

Greenberg, Richard

2011-03-01

Europa has become a high-priority objective for exploration because it may harbor life. Strategic planning for its exploration has been predicated on an extreme model in which the expected oceanic biosphere lies under a thick ice crust, buried too deep to be reached in the foreseeable future, which would beg the question of whether other active satellites might be more realistic objectives. However, Europa's ice may in fact be permeable, with very different implications for the possibilities for life and for mission planning. A biosphere may extend up to near the surface, making life far more readily accessible to exploration while at the same time making it vulnerable to contamination. The chances of finding life on Europa are substantially improved while the need for planetary protection becomes essential. The new National Research Council planetary protection study will need to go beyond its current mandate if meaningful standards are to be put in place. © Mary Ann Liebert, Inc.

RAS screening in colorectal cancer: a comprehensive analysis of the results from the UK NEQAS colorectal cancer external quality assurance schemes (2009-2016).

PubMed

Richman, Susan D; Fairley, Jennifer; Butler, Rachel; Deans, Zandra C

2017-12-01

Evidence strongly indicates that extended RAS testing should be undertaken in mCRC patients, prior to prescribing anti-EGFR therapies. With more laboratories implementing testing, the requirement for External Quality Assurance schemes increases, thus ensuring high standards of molecular analysis. Data was analysed from 15 United Kingdom National External Quality Assessment Service (UK NEQAS) for Molecular Genetics Colorectal cancer external quality assurance (EQA) schemes, delivered between 2009 and 2016. Laboratories were provided annually with nine colorectal tumour samples for genotyping. Information on methodology and extent of testing coverage was requested, and scores given for genotyping, interpretation and clerical accuracy. There has been a sixfold increase in laboratory participation (18 in 2009 to 108 in 2016). For RAS genotyping, fewer laboratories now use Roche cobas®, pyrosequencing and Sanger sequencing, with more moving to next generation sequencing (NGS). NGS is the most commonly employed technology for BRAF and PIK3CA mutation screening. KRAS genotyping errors were seen in ≤10% laboratories, until the 2014-2015 scheme, when there was an increase to 16.7%, corresponding to a large increase in scheme participants. NRAS genotyping errors peaked at 25.6% in the first 2015-2016 scheme but subsequently dropped to below 5%. Interpretation and clerical accuracy scores have been consistently good throughout. Within this EQA scheme, we have observed that the quality of molecular analysis for colorectal cancer has continued to improve, despite changes in the required targets, the volume of testing and the technologies employed. It is reassuring to know that laboratories clearly recognise the importance of participating in EQA schemes.

Assessment of Accuracy of Identification of Pathogenic Yeasts in Microbiology Laboratories in the United Kingdom

PubMed Central

Szekely, Adrien; Palmer, Michael D.; Johnson, Elizabeth M.

2012-01-01

Rapid, accurate identification of yeast isolates from clinical samples has always been important given their innately variable antifungal susceptibility profiles. Recently, this has become paramount with the proposed introduction of species-specific interpretive breakpoints for MICs obtained in yeast antifungal susceptibility tests (M. A. Pfaller, D. Andes, D. J. Diekema, A. Espinel–Ingroff, D. Sheehan, and CLSI Subcommittee for Antifungal Susceptibility Testing, Drug Resist. Updat. 13:180–195, 2010). Here, we present the results of a 12-month evaluation of the accuracy of identifications that accompany yeast isolates submitted to the Mycology Reference Laboratory (United Kingdom) for either confirmation of identity or susceptibility testing. In total, 1,781 yeast isolates were analyzed, and the robustness of prior identifications obtained in microbiology laboratories throughout the United Kingdom was assessed using a combination of culture on chromogenic agar, morphology on cornmeal agar, and molecular identification by pyrosequencing. Over 40% of isolates (755) were submitted without any suggested identification. Of those isolates with a prior identification, 100 (9.7%) were incorrectly identified. Error rates ranged from 5.2% (for organisms submitted for antifungal susceptibility testing) to 18.2% (for organisms requiring confirmation of identity) and varied in a strictly species-specific manner. At least 50% of identification errors would be likely to affect interpretation of MIC data, with a possible impact on patient management. In addition, 2.3% of submitted cultures were found to contain mixtures of at least two yeast species. The vast majority of mixtures had gone undetected in the referring laboratory and would have impacted the interpretation of antifungal susceptibility profiles and patient management. Some of the more common misidentifications are discussed according to the identification method employed, with suggestions for avoiding such misinterpretations. PMID:22649009

Assessment of accuracy of identification of pathogenic yeasts in microbiology laboratories in the United kingdom.

PubMed

Borman, Andrew M; Szekely, Adrien; Palmer, Michael D; Johnson, Elizabeth M

2012-08-01

Rapid, accurate identification of yeast isolates from clinical samples has always been important given their innately variable antifungal susceptibility profiles. Recently, this has become paramount with the proposed introduction of species-specific interpretive breakpoints for MICs obtained in yeast antifungal susceptibility tests (M. A. Pfaller, D. Andes, D. J. Diekema, A. Espinel-Ingroff, D. Sheehan, and CLSI Subcommittee for Antifungal Susceptibility Testing, Drug Resist. Updat. 13:180-195, 2010). Here, we present the results of a 12-month evaluation of the accuracy of identifications that accompany yeast isolates submitted to the Mycology Reference Laboratory (United Kingdom) for either confirmation of identity or susceptibility testing. In total, 1,781 yeast isolates were analyzed, and the robustness of prior identifications obtained in microbiology laboratories throughout the United Kingdom was assessed using a combination of culture on chromogenic agar, morphology on cornmeal agar, and molecular identification by pyrosequencing. Over 40% of isolates (755) were submitted without any suggested identification. Of those isolates with a prior identification, 100 (9.7%) were incorrectly identified. Error rates ranged from 5.2% (for organisms submitted for antifungal susceptibility testing) to 18.2% (for organisms requiring confirmation of identity) and varied in a strictly species-specific manner. At least 50% of identification errors would be likely to affect interpretation of MIC data, with a possible impact on patient management. In addition, 2.3% of submitted cultures were found to contain mixtures of at least two yeast species. The vast majority of mixtures had gone undetected in the referring laboratory and would have impacted the interpretation of antifungal susceptibility profiles and patient management. Some of the more common misidentifications are discussed according to the identification method employed, with suggestions for avoiding such misinterpretations.

An Investigation of the Critical Events and Influential Factors to the Evolution of the U.S. Man and the Biosphere Program

NASA Astrophysics Data System (ADS)

Thomsen, Jennifer M.

2018-04-01

The UNESCO Man and the Biosphere program has operated for 45 years as an international program that started in the 1970s to conserve biotic communities and provide areas for research, education, and training. The program later evolved in the 1990s to address social and environmental issues in a sustainable manner across a landscape. This program was one of the first efforts that recognized the importance of working beyond park and protected area boundaries and the need to sustain livelihoods as much as the resources. In the MAB program's infancy, the United States (U.S.) was a major advocate and leader with more than 45 biosphere reserves, most of them established in or around 1976. Yet, many political, economic, and other external factors influenced the U.S. MAB involvement in subsequent years. Consequently, the U.S. has remained largely inactive in the international MAB network for the past fifteen years until a recent push to revive the program under the leadership of the State Department and the National Park Service. Through in-depth research on two longterm U.S. biosphere reserves, this paper provides a description of the key events impacting the U.S. MAB program over the past several decades and discusses the influential role of politics, a public image, and the perceptions of international designations. Through the lessons presented in this paper, recommendations are provided to support the revival of the MAB program in the U.S.

The deep, hot biosphere: Twenty-five years of retrospection.

PubMed

Colman, Daniel R; Poudel, Saroj; Stamps, Blake W; Boyd, Eric S; Spear, John R

2017-07-03

Twenty-five years ago this month, Thomas Gold published a seminal manuscript suggesting the presence of a "deep, hot biosphere" in the Earth's crust. Since this publication, a considerable amount of attention has been given to the study of deep biospheres, their role in geochemical cycles, and their potential to inform on the origin of life and its potential outside of Earth. Overwhelming evidence now supports the presence of a deep biosphere ubiquitously distributed on Earth in both terrestrial and marine settings. Furthermore, it has become apparent that much of this life is dependent on lithogenically sourced high-energy compounds to sustain productivity. A vast diversity of uncultivated microorganisms has been detected in subsurface environments, and we show that H 2 , CH 4 , and CO feature prominently in many of their predicted metabolisms. Despite 25 years of intense study, key questions remain on life in the deep subsurface, including whether it is endemic and the extent of its involvement in the anaerobic formation and degradation of hydrocarbons. Emergent data from cultivation and next-generation sequencing approaches continue to provide promising new hints to answer these questions. As Gold suggested, and as has become increasingly evident, to better understand the subsurface is critical to further understanding the Earth, life, the evolution of life, and the potential for life elsewhere. To this end, we suggest the need to develop a robust network of interdisciplinary scientists and accessible field sites for long-term monitoring of the Earth's subsurface in the form of a deep subsurface microbiome initiative.

Probabilistic biosphere modeling for the long-term safety assessment of geological disposal facilities for radioactive waste using first- and second-order Monte Carlo simulation.

PubMed

Ciecior, Willy; Röhlig, Klaus-Jürgen; Kirchner, Gerald

2018-10-01

In the present paper, deterministic as well as first- and second-order probabilistic biosphere modeling approaches are compared. Furthermore, the sensitivity of the influence of the probability distribution function shape (empirical distribution functions and fitted lognormal probability functions) representing the aleatory uncertainty (also called variability) of a radioecological model parameter as well as the role of interacting parameters are studied. Differences in the shape of the output distributions for the biosphere dose conversion factor from first-order Monte Carlo uncertainty analysis using empirical and fitted lognormal distribution functions for input parameters suggest that a lognormal approximation is possibly not always an adequate representation of the aleatory uncertainty of a radioecological parameter. Concerning the comparison of the impact of aleatory and epistemic parameter uncertainty on the biosphere dose conversion factor, the latter here is described using uncertain moments (mean, variance) while the distribution itself represents the aleatory uncertainty of the parameter. From the results obtained, the solution space of second-order Monte Carlo simulation is much larger than that from first-order Monte Carlo simulation. Therefore, the influence of epistemic uncertainty of a radioecological parameter on the output result is much larger than that one caused by its aleatory uncertainty. Parameter interactions are only of significant influence in the upper percentiles of the distribution of results as well as only in the region of the upper percentiles of the model parameters. Copyright © 2018 Elsevier Ltd. All rights reserved.

Biospheric Cooling and the Emergence of Intelligence

NASA Astrophysics Data System (ADS)

Schwartzman, David; Middendorf, George

The long-term cooling history of the Earth's biosphere implies a temperature constraint on the timing of major events in biologic evolution, e.g., emergence of cyanobacteria, eucaryotes and Metazoa apparently occurred at times when temperatures were near their upper growth limits. Could biospheric cooling also have been a necessary condition for the emergence of veterbrates and their encephalization? The upper temperature limit for vertebrate growth is about 10 degrees below the limit for Metazoa (50 degrees C). Heterothermy followed by full homeothermy was likely a necessary condition for greater encephalization because of the energy requirement of larger brains. The temperature differential between an animal and a cooler environment, all other factors equal, will increase the efficiency of heat loss from the brain, but too large a differential will shift metabolic energy away from the brain to the procurement of food. Encephalization has also entailed the evolution of internal cooling mechanisms to avoid overheating the brain. The two periods of pronounced Phanerozoic cooling, the PermoCarboniferous and late Cenozoic, corresponded to the emergence of mammal-like reptiles and hominids respectively, with a variety of explanations offered for the apparent link. The origin of highly encephalized whales, dolphins and porpoises occurred with the drop in ocean temperatures 25-30 mya. Of course, other possible paths to encephalization are conceivable, with radically different solutions to the problem of heat dissipation. But the intrinsic requirements for information processing capacity necessary for intelligence suggest our terrestrial pattern may resemble those of alien biospheres given similar histories.

Value orientation and payment for ecosystem services: Perceived detrimental consequences lead to willingness-to-pay for ecosystem services.

PubMed

Obeng, Elizabeth Asantewaa; Aguilar, Francisco Xavier

2018-01-15

This research analyzed whether the three distinct value orientations posited under the Value-Belief-Norm (VBN) model determine willingness-to-pay (WTP) for a payment for ecosystem services (PES) program. A survey instrument gathered U.S. residents' knowledge and attitudes toward ecosystem services and PES, and elicited WTP for the restoration of a hypothetical degraded forest watershed for improved ecosystem services. Data from over 1000 respondents nationwide were analyzed using exploratory factor analysis (EFA) and ordered logistic regression. Urban respondents were more familiar with the concepts of ecosystem service and PES than rural respondents but familiarity did not yield statistically different WTP estimates. Based on results from the EFA, we posit that latent value orientations might be distinguished as 'detrimental', 'biospheric' and 'beneficial (egoistic)' - as compared to 'altruistic', 'biospheric' and 'egoistic' as suggested in the VBN's general awareness of consequences scale. Awareness of biospheric and detrimental consequences along with ascriptions to personal norms had positive and significant effects on stated WTP. Beneficial (egoistic) value orientation was negatively associated with WTP and carried a negative average WTP per household per year (US$ -30.48) for the proposed PES restoration program as compared with biospheric (US$ 15.53) and detrimental (US$ 3.96) orientations. Besides personal norms, awareness of detrimental consequences to human wellbeing from environmental degradation seems the stronger driver of WTP for the restoration and protection of forest watershed ecosystem services under a PES program. Copyright © 2017 Elsevier Ltd. All rights reserved.

An Investigation of the Critical Events and Influential Factors to the Evolution of the U.S. Man and the Biosphere Program.

PubMed

Thomsen, Jennifer M

2018-04-01

The UNESCO Man and the Biosphere program has operated for 45 years as an international program that started in the 1970s to conserve biotic communities and provide areas for research, education, and training. The program later evolved in the 1990s to address social and environmental issues in a sustainable manner across a landscape. This program was one of the first efforts that recognized the importance of working beyond park and protected area boundaries and the need to sustain livelihoods as much as the resources. In the MAB program's infancy, the United States (U.S.) was a major advocate and leader with more than 45 biosphere reserves, most of them established in or around 1976. Yet, many political, economic, and other external factors influenced the U.S. MAB involvement in subsequent years. Consequently, the U.S. has remained largely inactive in the international MAB network for the past fifteen years until a recent push to revive the program under the leadership of the State Department and the National Park Service. Through in-depth research on two longterm U.S. biosphere reserves, this paper provides a description of the key events impacting the U.S. MAB program over the past several decades and discusses the influential role of politics, a public image, and the perceptions of international designations. Through the lessons presented in this paper, recommendations are provided to support the revival of the MAB program in the U.S.

Snowball Earth: Response of the biosphere?

NASA Astrophysics Data System (ADS)

Runnegar, B.

2001-05-01

Snowball Earth is a script for global catastrophe that rivals giant impact theories in the likely severity of its environmental effects. This is particularly true for the "hard" version of the hypothesis, which requires the atmosphere to be effectively isolated from the ocean so that its carbon dioxide concentration can build up to the level ( ~100 PAL) ultimately required to melt the ice. However, coupled GCM-EMB models (Hyde et al. Nature 405, 425-430; Crowley & Hyde, GRL 28, 283-286) allow equatorial open water solutions under plausible Neoproterozoic conditions. These "softer" scenarios are more appealing if one considers the possible effects of snowball Earth episodes on the global biosphere. The meager Neoproterozoic fossil record makes it difficult to observe the biospheric response directly, but we know from evolutionary trees constructed from aligned protein and DNA sequences from living organisms, calibrated by the fossil record, that many lines of descent passed through the Cryogenian glacial periods. They include various kinds of prokaryotic and eukaryotic algae, a range of protists, and probably, a number of different kinds of animals and fungi. In addition, most of the microbial groups shown on comprehensive 16S rRNA trees have molecular clock ages that predate the snowball episodes. As the global environmental perturbations associated with the "hard" snowball hypothesis (freezing temperatures; huge and rapid changes in temperature; sudden carbon dioxide overload) are thought to have been biologically limiting during the Phanerozoic, the inferred response of the biosphere to Neoprotereozic glaciations may, indeed, provide a way of testing alternative snowball Earth scenarios.

Human impact on the planet: an earth system science perspective and ethical considerations

USGS Publications Warehouse

Williams, Richard S.

2002-01-01

The modern Earth Narrative, the scientific story of the 4.5 billion-year natural and human history of the Earth, has emerged from the solid foundation of two factual concepts: Deep (or Geologic) Time and Biological Evolution. spread acceptance of the Earth Narrative is critically important as we begin the third millennium, because it provides a clear understanding of the growing impact of human population growth and associated activities on the Earth System, especially the negative impact on Earth?s biosphere. It is important for humans to realize that we are but one of 4,500 species of mammals that exist on Earth and that we are but one species in the estimated 30 to 100 million species that form the complex biosphere. We also need to recognize that all species exist within the physical limits imposed by the geosphere. We are totally dependent on the biosphere for food, oxygen, and other necessities of life. mans are one of the latest results of biological evolution operating over a long period of Geologic Time. We find ourselves on Earth, after 4.5 billion years of Earth history by chance, not by design. Humans have become so successful at modifying their environment that many of the natural limitations on the expansion of populations of our fellow animals have been overcome by technological and cultural innovations. According to Peter Raven, ?Humans, at a current population of 6 billion [expected to nearly double by 2050], are consuming or wasting about 50 percent of the total net biological productivity on land and 50 percent of the available supply of freshwater. The overwhelming and expanding human presence leaves less and less room in the environment for other biota.? st century will be a pivotal time in the fate of Earth?s biosphere. Whereas human modification of the geosphere will slowly recover over time, human changes to the biosphere are a far more consequential matter? extinction of a species is forever! Will humans effectively use our new knowledge of natural and human history to stop further degradation of Earth?s ecosystems and extinction of its biota? The fate of the biosphere, including humanity, depends on a reaffirmation by all humans of all cultures and religions of the global importance of a planet-wide conservation of the Earth?s biotic heritage. For the world?s religions it means elevation of stewardship of the Earth to a moral imperative and a goal of complete preservation of the Earth?s biotic inheritance, one which is based on a Do No Harm ethic.

Using observations to evaluate biosphere-atmosphere interactions in models

NASA Astrophysics Data System (ADS)

Green, Julia; Konings, Alexandra G.; Alemohammad, Seyed H.; Gentine, Pierre

2017-04-01

Biosphere-atmosphere interactions influence the hydrologic cycle by altering climate and weather patterns (Charney, 1975; Koster et al., 2006; Seneviratne et al., 2006), contributing up to 30% of precipitation and radiation variability in certain regions (Green et al., 2017). They have been shown to contribute to the persistence of drought in Europe (Seneviratne et al., 2006), as well as to increase rainfall in the Amazon (Spracklen et al., 2012). Thus, a true representation of these feedbacks in Earth System Models (ESMs) is crucial for accurate forecasting and planning. However, it has been difficult to validate the performance of ESMs since often-times surface and atmospheric flux data are scarce and/or difficult to observe. In this study, we use the results of a new global observational study (using remotely sensed solar-induced fluorescence to represent the biosphere flux) (Green et al., 2017) to determine how well a suite of 13 ESMs capture biosphere-atmosphere feedbacks. We perform a Conditional Multivariate Granger Causality analysis in the frequency domain with radiation, precipitation and temperature as atmospheric inputs and GPP as the biospheric input. Performing the analysis in the frequency domain allows for separation of feedbacks at different time-scales (subseasonal, seasonal or interannual). Our findings can be used to determine whether there is agreement between models, as well as, to pinpoint regions or time-scales of model bias or inaccuracy, which will provide insight on potential improvement. We demonstrate that in addition to the well-known problem of convective parameterization over land in models, the main issue in representing feedbacks between the land and the atmosphere is due to the misrepresentation of water stress. These results provide a direct quantitative assessment of feedbacks in models and how to improve them. References: Charney, J.G. Dynamics of deserts and drought in the Sahel. Quarterly Journal of the Royal Meteorological Society 101, 193-202 (1975). Green, J.K., Konings, A.G., Alemohammad, S.H., Berry, J., Entekhabi, D., Kolassa, J., Lee, J.E., Gentine, P. Hotspots of terrestrial biosphere-atmosphere feedbacks. In review. (2017). Koster, R. D. et al. GLACE: The Global Land - Atmosphere Coupling Experiment. Part I: Overview. J. Hydrometeorol. 7, 611-625 (2006). Seneviratne, S.I., Lüthi, D., Litschi M., Schär, C. Land-atmosphere coupling and climate change in Europe. Nature 443, 205-209 (2006). Spracklen, D.V, Arnold, S.R., Taylor, C.M. Observations of increased tropical rainfall preceded by air passage over forests. Nature 489, 282-285 (2012).

Earth's Early Biosphere and the Biogeochemical Carbon Cycle

NASA Technical Reports Server (NTRS)

DesMarais, David

2004-01-01

Our biosphere has altered the global environment principally by influencing the chemistry of those elements most important for life, e g., C, N, S, O, P and transition metals (e.g., Fe and Mn). The coupling of oxygenic photosynthesis with the burial in sediments of photosynthetic organic matter, and with the escape of H2 to space, has increased the state of oxidation of the Oceans and atmosphere. It has also created highly reduced conditions within sedimentary rocks that have also extensively affected the geochemistry of several elements. The decline of volcanism during Earth's history reduced the flow of reduced chemical species that reacted with photosynthetically produced O2. The long-term net accumulation of photosynthetic O2 via biogeochemical processes has profoundly influenced our atmosphere and biosphere, as evidenced by the O2 levels required for algae, multicellular life and certain modem aerobic bacteria to exist. When our biosphere developed photosynthesis, it tapped into an energy resource that was much larger than the energy available from oxidation-reduction reactions associated with weathering and hydrothermal activity. Today, hydrothermal sources deliver globally (0.13-1.1)x10(exp l2) mol yr(sup -1) of reduced S, Fe(2+), Mn(2+), H2 and CH4; this is estimated to sustain at most about (0.2-2)xl0(exp 12)mol C yr(sup -1) of organic carbon production by chemautotrophic microorganisms. In contrast, global photosynthetic productivity is estimated to be 9000x10(exp 12) mol C yr(sup -1). Thus, even though global thermal fluxes were greater in the distant geologic past than today, the onset of oxygenic photosynthesis probably increased global organic productivity by some two or more orders of magnitude. This enormous productivity materialized principally because oxygenic photosynthesizers unleashed a virtually unlimited supply of reduced H that forever freed life from its sole dependence upon abiotic sources of reducing power such as hydrothermal emanations and weathering. Communities sustained by oxygenic photosynthesis apparently thrived wherever supplies of sunlight, moisture and nutrients were sufficient. Prior to the development of oxygenic photosynthesis, the net global effect of the ancient global biosphere was to facilitate chemical equilibrium between reduced species from thermal activity and weathering and more oxidized constituents in the surface environment. But even this ancient biosphere might have been globally pervasive. The global geothermal heat flow was substantially higher during Earth's first billion years, and thus reduced chemical species might have persisted in sunlit aquatic environments. Perhaps the substantial decline in thermal activity between 4000 and 3000 Ma created a driver for oxygenic photosynthesis to develop.

Pyrosequencing assessment of rhizosphere fungal communities from a soybean field.

PubMed

Sugiyama, Akifumi; Ueda, Yoshikatsu; Takase, Hisabumi; Yazaki, Kazufumi

2014-10-01

Soil fungal communities play essential roles in soil ecosystems, affecting plant growth and health. Rhizosphere bacterial communities have been shown to undergo dynamic changes during plant growth. This study utilized 454 pyrosequencing to analyze rhizosphere fungal communities during soybean growth. Members of the Ascomycota and Basiodiomycota dominated in all soils. There were no statistically significant changes at the phylum level among growth stages or between bulk and rhizosphere soils. In contrast, the relative abundance of small numbers of operational taxonomic units, 4 during growth and 28 between bulk and rhizosphere soils, differed significantly. Clustering analysis revealed that rhizosphere fungal communities were different from bulk fungal communities during growth stages of soybeans. Taken together, these results suggest that in contrast to rhizosphere bacterial communities, most constituents of rhizosphere fungal communities remained stable during soybean growth.

FT-IR spectroscopy: A powerful tool for studying the inter- and intraspecific biodiversity of cultivable non-Saccharomyces yeasts isolated from grape must.

PubMed

Grangeteau, Cédric; Gerhards, Daniel; Terrat, Sebastien; Dequiedt, Samuel; Alexandre, Hervé; Guilloux-Benatier, Michèle; von Wallbrunn, Christian; Rousseaux, Sandrine

2016-02-01

The efficiency of the FT-IR technique for studying the inter- and intra biodiversity of cultivable non-Saccharomyces yeasts (NS) present in different must samples was examined. In first, the capacity of the technique FT-IR to study the global diversity of a given sample was compared to the pyrosequencing method, used as a reference technique. Seven different genera (Aureobasidium, Candida, Cryptococcus, Hanseniaspora, Issatchenkia, Metschnikowia and Pichia) were identified by FT-IR and also by pyrosequencing. Thirty-eight other genera were identified by pyrosequencing, but together they represented less than 6% of the average total population of 6 musts. Among the species identified, some of them present organoleptic potentials in winemaking, particularly Starmerella bacillaris (synonym Candidazemplinina). So in a second time, we evaluated the capacity of the FT-IR technique to discriminate the isolates of this species because few techniques were able to study intraspecific NS yeast biodiversity. The results obtained were validated by using a classic method as ITS sequencing. Biodiversity at strain level was high: 19 different strains were identified from 58 isolates. So, FT-IR spectroscopy seems to be an accurate and reliable method for identifying major genera present in the musts. The two biggest advantages of the FT-IR are the capacity to characterize intraspecific biodiversity of non-Saccharomyces yeasts and the possibility to discriminate a lot of strains. Copyright © 2015 Elsevier B.V. All rights reserved.

A microbial spoilage profile of half shell Pacific oysters (Crassostrea gigas) and Sydney rock oysters (Saccostrea glomerata).

PubMed

Madigan, Thomas L; Bott, Nathan J; Torok, Valeria A; Percy, Nigel J; Carragher, John F; de Barros Lopes, Miguel A; Kiermeier, Andreas

2014-04-01

This study aimed to assess bacterial spoilage of half shell Pacific and Sydney rock oysters during storage using microbial culture and 16S rRNA pyrosequencing. Odour and pH of oyster meats were also investigated. Estimation of microbiological counts by microbial culture highlighted growth of psychrotrophic bacteria. During storage, odour scores (a score describing deterioration of fresh odours where a score of 1 is fresh and 4 is completely spoiled) increased from 1.0 to 3.0 for Pacific oysters and from 1.3 to 3.4 for Sydney rock oysters. pH results obtained for both species fluctuated during storage (range 6.28-6.73) with an overall increase at end of storage. Pyrosequencing revealed that the majority of bacteria at Day 0 represented taxa from amongst the Proteobacteria, Tenericutes and Spirochaetes that have not been cultured and systematically described. During storage, Proteobacteria became abundant with Pseudoalteromonas and Vibrio found to be dominant in both oyster species at Day 7. Analysis of the pyrosequencing data showed significant differences in bacterial profiles between oyster species and storage time (both P = 0.001). As oysters spoiled, bacterial profiles between oyster species became more similar indicating a common spoilage profile. Data presented here provides detailed insight into the changing bacterial profile of shucked oysters during storage and has identified two genera, Pseudoalteromonas and Vibrio, as being important in spoilage of shucked oysters. Copyright © 2013 Elsevier Ltd. All rights reserved.

Rapid quantitation of neuraminidase inhibitor drug resistance in influenza virus quasispecies.

PubMed

Lackenby, Angie; Democratis, Jane; Siqueira, Marilda M; Zambon, Maria C

2008-01-01

Emerging resistance of influenza viruses to neuraminidase inhibitors is a concern, both in surveillance of global circulating strains and in treatment of individual patients. Current methodologies to detect resistance rely on the use of cultured virus, thus taking time to complete or lacking the sensitivity to detect mutations in viral quasispecies. Methodology for rapid detection of clinically meaningful resistance is needed to assist individual patient management and to track the transmission of resistant viruses in the community. We have developed a pyrosequencing methodology to detect and quantitate influenza neuraminidase inhibitor resistance mutations in cultured virus and directly in clinical material. Our assays target polymorphisms associated with drug resistance in the neuraminidase genes of human influenza A H1N1 as well as human and avian H5N1 viruses. Quantitation can be achieved using viral RNA extracted directly from respiratory or tissue samples, thus eliminating the need for virus culture and allowing the assay of highly pathogenic viruses such as H5N1 without high containment laboratory facilities. Antiviral-resistant quasispecies are detected and quantitated accurately when present in the total virus population at levels as low as 10%. Pyrosequencing is a real-time assay; therefore, results can be obtained within a clinically relevant timeframe and provide information capable of informing individual patient or outbreak management. Pyrosequencing is ideally suited for early identification of emerging antiviral resistance in human and avian influenza infection and is a useful tool for laboratory surveillance and pandemic preparedness.

Impact of subacute ruminal acidosis (SARA) adaptation on rumen microbiota in dairy cattle using pyrosequencing.

PubMed

Mao, S Y; Zhang, R Y; Wang, D S; Zhu, W Y

2013-12-01

The objective of this study was to evaluate the changes in bacterial populations in the rumen of dairy cattle following adaptation to subacute ruminal acidosis (SARA) using 16S rRNA gene pyrosequencing. Rumen contents were collected from four cattle adapted to either a 40% (control diet, COD) or 70% (SARA induction diet, SAID) concentrate feeds. DNA was extracted from each of the samples. Bacterial 16S rRNA genes of ruminal DNA extracts were PCR amplified with 2 bar coded primer sets and sequenced by 454 pyrosequencing. At a high taxonomic level, the percentage of Proteobacteria and Bacteroidetes were reduced by SAID feeding, whereas Firmicutes and Actinobacteria were more abundant in the SAID than in the COD group. At the genus level, as compared with the COD group, the abundances of Prevotella, Treponema, Anaeroplasma, Papillibacter, Acinetobacter and unclassified populations including unclassified Lentisphaerae, and unclassified bacteria were lower (P < 0.05), while the percentages of Ruminococcus, Atopobium, unclassified Clostridiales and Bifidobacterium were increased (P < 0.05) in the SAID group. Feeding of SAID reduced (P < 0.001) the diversity of the rumen microbial community. Taken together, our findings provide a comprehensive picture of current knowledge of the community structure of the rumen bacterial ecosystem during SARA, and enhance our understanding about the ruminal microbial ecology that may be useful in the prevention of ruminal acidosis. Copyright © 2013 Elsevier Ltd. All rights reserved.

Comparisons of the fungal and protistan communities among different marine sponge holobionts by pyrosequencing.

PubMed

He, Liming; Liu, Fang; Karuppiah, Valliappan; Ren, Yi; Li, Zhiyong

2014-05-01

To date, the knowledge of eukaryotic communities associated with sponges remains limited compared with prokaryotic communities. In a manner similar to prokaryotes, it could be hypothesized that sponge holobionts have phylogenetically diverse eukaryotic symbionts, and the eukaryotic community structures in different sponge holobionts were probably different. In order to test this hypothesis, the communities of eukaryota associated with 11 species of South China Sea sponges were compared with the V4 region of 18S ribosomal ribonucleic acid gene using 454 pyrosequencing. Consequently, 135 and 721 unique operational taxonomic units (OTUs) of fungi and protists were obtained at 97 % sequence similarity, respectively. These sequences were assigned to 2 phyla of fungi (Ascomycota and Basidiomycota) and 9 phyla of protists including 5 algal phyla (Chlorophyta, Haptophyta, Streptophyta, Rhodophyta, and Stramenopiles) and 4 protozoal phyla (Alveolata, Cercozoa, Haplosporidia, and Radiolaria) including 47 orders (12 fungi, 35 protists). Entorrhizales of fungi and 18 orders of protists were detected in marine sponges for the first time. Particularly, Tilletiales of fungi and Chlorocystidales of protists were detected for the first time in marine habitats. Though Ascomycota, Alveolata, and Radiolaria were detected in all the 11 sponge species, sponge holobionts have different fungi and protistan communities according to OTU comparison and principal component analysis at the order level. This study provided the first insights into the fungal and protistan communities associated with different marine sponge holobionts using pyrosequencing, thus further extending the knowledge on sponge-associated eukaryotic diversity.

Bacterial identification and subtyping using DNA microarray and DNA sequencing.

PubMed

Al-Khaldi, Sufian F; Mossoba, Magdi M; Allard, Marc M; Lienau, E Kurt; Brown, Eric D

2012-01-01

The era of fast and accurate discovery of biological sequence motifs in prokaryotic and eukaryotic cells is here. The co-evolution of direct genome sequencing and DNA microarray strategies not only will identify, isotype, and serotype pathogenic bacteria, but also it will aid in the discovery of new gene functions by detecting gene expressions in different diseases and environmental conditions. Microarray bacterial identification has made great advances in working with pure and mixed bacterial samples. The technological advances have moved beyond bacterial gene expression to include bacterial identification and isotyping. Application of new tools such as mid-infrared chemical imaging improves detection of hybridization in DNA microarrays. The research in this field is promising and future work will reveal the potential of infrared technology in bacterial identification. On the other hand, DNA sequencing by using 454 pyrosequencing is so cost effective that the promise of $1,000 per bacterial genome sequence is becoming a reality. Pyrosequencing technology is a simple to use technique that can produce accurate and quantitative analysis of DNA sequences with a great speed. The deposition of massive amounts of bacterial genomic information in databanks is creating fingerprint phylogenetic analysis that will ultimately replace several technologies such as Pulsed Field Gel Electrophoresis. In this chapter, we will review (1) the use of DNA microarray using fluorescence and infrared imaging detection for identification of pathogenic bacteria, and (2) use of pyrosequencing in DNA cluster analysis to fingerprint bacterial phylogenetic trees.

Global characterization of Artemisia annua glandular trichome transcriptome using 454 pyrosequencing

PubMed Central

Wang, Wei; Wang, Yejun; Zhang, Qing; Qi, Yan; Guo, Dianjing

2009-01-01

Background Glandular trichomes produce a wide variety of commercially important secondary metabolites in many plant species. The most prominent anti-malarial drug artemisinin, a sesquiterpene lactone, is produced in glandular trichomes of Artemisia annua. However, only limited genomic information is currently available in this non-model plant species. Results We present a global characterization of A. annua glandular trichome transcriptome using 454 pyrosequencing. Sequencing runs using two normalized cDNA collections from glandular trichomes yielded 406,044 expressed sequence tags (average length = 210 nucleotides), which assembled into 42,678 contigs and 147,699 singletons. Performing a second sequencing run only increased the number of genes identified by ~30%, indicating that massively parallel pyrosequencing provides deep coverage of the A. annua trichome transcriptome. By BLAST search against the NCBI non-redundant protein database, putative functions were assigned to over 28,573 unigenes, including previously undescribed enzymes likely involved in sesquiterpene biosynthesis. Comparison with ESTs derived from trichome collections of other plant species revealed expressed genes in common functional categories across different plant species. RT-PCR analysis confirmed the expression of selected unigenes and novel transcripts in A. annua glandular trichomes. Conclusion The presence of contigs corresponding to enzymes for terpenoids and flavonoids biosynthesis suggests important metabolic activity in A. annua glandular trichomes. Our comprehensive survey of genes expressed in glandular trichome will facilitate new gene discovery and shed light on the regulatory mechanism of artemisinin metabolism and trichome function in A. annua. PMID:19818120

KRAS Mutation Test in Korean Patients with Colorectal Carcinomas: A Methodological Comparison between Sanger Sequencing and a Real-Time PCR-Based Assay.

PubMed

Lee, Sung Hak; Chung, Arthur Minwoo; Lee, Ahwon; Oh, Woo Jin; Choi, Yeong Jin; Lee, Youn-Soo; Jung, Eun Sun

2017-01-01

Mutations in the KRAS gene have been identified in approximately 50% of colorectal cancers (CRCs). KRAS mutations are well established biomarkers in anti-epidermal growth factor receptor therapy. Therefore, assessment of KRAS mutations is needed in CRC patients to ensure appropriate treatment. We compared the analytical performance of the cobas test to Sanger sequencing in 264 CRC cases. In addition, discordant specimens were evaluated by 454 pyrosequencing. KRAS mutations for codons 12/13 were detected in 43.2% of cases (114/264) by Sanger sequencing. Of 257 evaluable specimens for comparison, KRAS mutations were detected in 112 cases (43.6%) by Sanger sequencing and 118 cases (45.9%) by the cobas test. Concordance between the cobas test and Sanger sequencing for each lot was 93.8% positive percent agreement (PPA) and 91.0% negative percent agreement (NPA) for codons 12/13. Results from the cobas test and Sanger sequencing were discordant for 20 cases (7.8%). Twenty discrepant cases were subsequently subjected to 454 pyrosequencing. After comprehensive analysis of the results from combined Sanger sequencing-454 pyrosequencing and the cobas test, PPA was 97.5% and NPA was 100%. The cobas test is an accurate and sensitive test for detecting KRAS -activating mutations and has analytical power equivalent to Sanger sequencing. Prescreening using the cobas test with subsequent application of Sanger sequencing is the best strategy for routine detection of KRAS mutations in CRC.

Diversity patterns and activity of uncultured marine heterotrophic flagellates unveiled with pyrosequencing

PubMed Central

Logares, Ramiro; Audic, Stephane; Santini, Sebastien; Pernice, Massimo C; de Vargas, Colomban; Massana, Ramon

2012-01-01

Flagellated heterotrophic microeukaryotes have key roles for the functioning of marine ecosystems as they channel large amounts of organic carbon to the upper trophic levels and control the population sizes of bacteria and archaea. Still, we know very little on the diversity patterns of most groups constituting this evolutionary heterogeneous assemblage. Here, we investigate 11 groups of uncultured flagellates known as MArine STramenopiles (MASTs). MASTs are ecologically very important and branch at the base of stramenopiles. We explored the diversity patterns of MASTs using pyrosequencing (18S rDNA) in coastal European waters. We found that MAST groups range from highly to lowly diversified. Pyrosequencing (hereafter ‘454') allowed us to approach to the limits of taxonomic diversity for all MAST groups, which varied in one order of magnitude (tens to hundreds) in terms of operational taxonomic units (98% similarity). We did not evidence large differences in activity, as indicated by ratios of DNA:RNA-reads. Most groups were strictly planktonic, although we found some groups that were active in sediments and even in anoxic waters. The proportion of reads per size fraction indicated that most groups were composed of very small cells (∼2–5 μm). In addition, phylogenetically different assemblages appeared to be present in different size fractions, depths and geographic zones. Thus, MAST diversity seems to be highly partitioned in spatial scales. Altogether, our results shed light on these ecologically very important but poorly known groups of uncultured marine flagellates. PMID:22534609

Pyrosequencing of prey DNA in reptile faeces: analysis of earthworm consumption by slow worms.

PubMed

Brown, David S; Jarman, Simon N; Symondson, William O C

2012-03-01

Little quantitative ecological information exists on the diets of most invertebrate feeding reptiles, particularly nocturnal or elusive species that are difficult to observe. In the UK and elsewhere, reptiles are legally required to be relocated before land development can proceed, but without knowledge of their dietary requirements, the suitability of receptor sites cannot be known. Here, we tested the ability of non-invasive DNA-based molecular diagnostics (454 pyrosequencing) to analyse reptile diets, with the specific aims of determining which earthworm species are exploited by slow worms (the legless lizard Anguis fragilis) and whether they feed on the deeper-living earthworm species that only come to the surface at night. Slow worm faecal samples from four different habitats were analysed using earthworm-specific PCR primers. We found that 86% of slow worms (N=80) had eaten earthworms. In lowland heath and marshy/acid grassland, Lumbricus rubellus, a surface-dwelling epigeic species, dominated slow worm diet. In two other habitats, riverside pasture and calciferous coarse grassland, diet was dominated by deeper-living anecic and endogeic species. We conclude that all species of earthworm are exploited by these reptiles and lack of specialization allows slow worms to thrive in a wide variety of habitats. Pyrosequencing of prey DNA in faeces showed promise as a practical, rapid and relatively inexpensive means of obtaining detailed and valuable ecological information on the diets of reptiles. © 2011 Blackwell Publishing Ltd.

Alignment-free inference of hierarchical and reticulate phylogenomic relationships.

PubMed

Bernard, Guillaume; Chan, Cheong Xin; Chan, Yao-Ban; Chua, Xin-Yi; Cong, Yingnan; Hogan, James M; Maetschke, Stefan R; Ragan, Mark A

2017-06-30

We are amidst an ongoing flood of sequence data arising from the application of high-throughput technologies, and a concomitant fundamental revision in our understanding of how genomes evolve individually and within the biosphere. Workflows for phylogenomic inference must accommodate data that are not only much larger than before, but often more error prone and perhaps misassembled, or not assembled in the first place. Moreover, genomes of microbes, viruses and plasmids evolve not only by tree-like descent with modification but also by incorporating stretches of exogenous DNA. Thus, next-generation phylogenomics must address computational scalability while rethinking the nature of orthogroups, the alignment of multiple sequences and the inference and comparison of trees. New phylogenomic workflows have begun to take shape based on so-called alignment-free (AF) approaches. Here, we review the conceptual foundations of AF phylogenetics for the hierarchical (vertical) and reticulate (lateral) components of genome evolution, focusing on methods based on k-mers. We reflect on what seems to be successful, and on where further development is needed. © The Author 2017. Published by Oxford University Press.

Orthoptera (Insecta: Tettigonioidea, Pyrgomorphoidea, Acridoidea) of Kafa Biosphere Reserve, Bale Mountains National Park and other areas of conservation interest in Ethiopia.

PubMed

Felix, Rob P W H; Massa, Bruno

2016-11-09

An annotated checklist of 51 Orthoptera taxa (Tettigonioidea, Pyrgomorphoidea and Acridoidea) of Kafa Biosphere Reserve, Bale Mountains National Park and some additional sites in Ethiopia is presented. Ten species are newly recorded for the country. Four species are new to science: Peropyrrhicia attilioi n. sp., P. keffensis n. sp., P. semiensis n. sp. and Coryphosima danieli n. sp. The status of Peropyrrhicia cooperi Uvarov, 1934 and P. scotti Uvarov, 1934 is revised: both are considered valid species.

The biosphere as a driver of global atmospheric change

NASA Technical Reports Server (NTRS)

Levine, Joel S.

1991-01-01

The effects of the biosphere on the evolution of atmospheric oxygen and ozone, and the consequences of that development for global atmospheric change, are discussed. Attention is given to the impact of oxygen and ozone on atmospheric photolysis rates, the effect of oxygen on the biogenic production of nitrous oxide and nitric oxide, and the effects of the evolution of atmospheric oxygen on fires and biomass burning. The influence of the latter on atmospheric processes, particularly the production of methane, carbon dioxide, and carbon monoxide, is considered.

Uncertainty analysis of the Operational Simplified Surface Energy Balance (SSEBop) model at multiple flux tower sites

USGS Publications Warehouse

Chen, Mingshi; Senay, Gabriel B.; Singh, Ramesh K.; Verdin, James P.

2016-01-01

Evapotranspiration (ET) is an important component of the water cycle – ET from the land surface returns approximately 60% of the global precipitation back to the atmosphere. ET also plays an important role in energy transport among the biosphere, atmosphere, and hydrosphere. Current regional to global and daily to annual ET estimation relies mainly on surface energy balance (SEB) ET models or statistical and empirical methods driven by remote sensing data and various climatological databases. These models have uncertainties due to inevitable input errors, poorly defined parameters, and inadequate model structures. The eddy covariance measurements on water, energy, and carbon fluxes at the AmeriFlux tower sites provide an opportunity to assess the ET modeling uncertainties. In this study, we focused on uncertainty analysis of the Operational Simplified Surface Energy Balance (SSEBop) model for ET estimation at multiple AmeriFlux tower sites with diverse land cover characteristics and climatic conditions. The 8-day composite 1-km MODerate resolution Imaging Spectroradiometer (MODIS) land surface temperature (LST) was used as input land surface temperature for the SSEBop algorithms. The other input data were taken from the AmeriFlux database. Results of statistical analysis indicated that the SSEBop model performed well in estimating ET with an R2 of 0.86 between estimated ET and eddy covariance measurements at 42 AmeriFlux tower sites during 2001–2007. It was encouraging to see that the best performance was observed for croplands, where R2 was 0.92 with a root mean square error of 13 mm/month. The uncertainties or random errors from input variables and parameters of the SSEBop model led to monthly ET estimates with relative errors less than 20% across multiple flux tower sites distributed across different biomes. This uncertainty of the SSEBop model lies within the error range of other SEB models, suggesting systematic error or bias of the SSEBop model is within the normal range. This finding implies that the simplified parameterization of the SSEBop model did not significantly affect the accuracy of the ET estimate while increasing the ease of model setup for operational applications. The sensitivity analysis indicated that the SSEBop model is most sensitive to input variables, land surface temperature (LST) and reference ET (ETo); and parameters, differential temperature (dT), and maximum ET scalar (Kmax), particularly during the non-growing season and in dry areas. In summary, the uncertainty assessment verifies that the SSEBop model is a reliable and robust method for large-area ET estimation. The SSEBop model estimates can be further improved by reducing errors in two input variables (ETo and LST) and two key parameters (Kmax and dT).

Temporal variations of atmospheric CO2 and CO at Ahmedabad in western India

NASA Astrophysics Data System (ADS)

Chandra, Naveen; Lal, Shyam; Venkataramani, S.; Patra, Prabir K.; Sheel, Varun

2016-05-01

About 70 % of the anthropogenic carbon dioxide (CO2) is emitted from the megacities and urban areas of the world. In order to draw effective emission mitigation policies for combating future climate change as well as independently validating the emission inventories for constraining their large range of uncertainties, especially over major metropolitan areas of developing countries, there is an urgent need for greenhouse gas measurements over representative urban regions. India is a fast developing country, where fossil fuel emissions have increased dramatically in the last three decades and are predicted to continue to grow further by at least 6 % per year through to 2025. The CO2 measurements over urban regions in India are lacking. To overcome this limitation, simultaneous measurements of CO2 and carbon monoxide (CO) have been made at Ahmedabad, a major urban site in western India, using a state-of-the-art laser-based cavity ring down spectroscopy technique from November 2013 to May 2015. These measurements enable us to understand the diurnal and seasonal variations in atmospheric CO2 with respect to its sources (both anthropogenic and biospheric) and biospheric sinks. The observed annual average concentrations of CO2 and CO are 413.0 ± 13.7 and 0.50 ± 0.37 ppm respectively. Both CO2 and CO show strong seasonality with lower concentrations (400.3 ± 6.8 and 0.19 ± 0.13 ppm) during the south-west monsoon and higher concentrations (419.6 ± 22.8 and 0.72 ± 0.68 ppm) during the autumn (SON) season. Strong diurnal variations are also observed for both the species. The common factors for the diurnal cycles of CO2 and CO are vertical mixing and rush hour traffic, while the influence of biospheric fluxes is also seen in the CO2 diurnal cycle. Using CO and CO2 covariation, we differentiate the anthropogenic and biospheric components of CO2 and found significant contributions of biospheric respiration and anthropogenic emissions in the late night (00:00-05:00 h, IST) and evening rush hours (18:00-22:00 h) respectively. We compute total yearly emissions of CO to be 69.2 ± 0.07 Gg for the study region using the observed CO : CO2 correlation slope and bottom-up CO2 emission inventory. This calculated emission of CO is 52 % larger than the estimated emission of CO by the emissions database for global atmospheric research (EDGAR) inventory. The observations of CO2 have been compared with an atmospheric chemistry-transport model (ACTM), which incorporates various components of CO2 fluxes. ACTM is able to capture the basic variabilities, but both diurnal and seasonal amplitudes are largely underestimated compared to the observations. We attribute this underestimation by the model to uncertainties in terrestrial biosphere fluxes and coarse model resolution. The fossil fuel signal from the model shows fairly good correlation with observed CO2 variations, which supports the overall dominance of fossil fuel emissions over the biospheric fluxes in this urban region.

Biogeosystem technique - the fundamental base of modern Water Policy and Management

NASA Astrophysics Data System (ADS)

Kalinitchenko, Valery; Batukaev, Abdulmalik; Minkina, Tatiana; Solntseva, Natalia; Skovpen, Andrey; Zarmaev, Ali; Jusupov, Vaha; Lohmanova, Olga

2014-05-01

Freshwater conservation is the problem of world water strategy. Water is intended not only for human consumption but also for functions of the biosphere - the only place where humanity can exist. To maintain the quality of biosphere is very relevant. An important property of biosphere is ability of soil to provide the synthesis of fresh living biological material by plants. There are few places in the world where exists a natural high level of biological production. Therefore, irrigation widely applies. Irrigation provides an increase of crops, but the imitating gravitational frontal isotropic-continual irrigation paradigm has the adverse effects on soils and landscapes. So irrigation in the past history of humanity was one of the causes for civilization's downfall, Sumer in particular, now irrigation causes a humanitarian catastrophe in Central Asia. Irrigation is the world main consumer of water. Leading cause of negative results of irrigation in biosphere is the irrigation paradigm defect. By artificial watering is imitated a natural hydrological regime of the land. The water flows down into soil through the soil surface. Or groundwater flows up through the soil bottom. In either case, a natural or standard artificial soil moisturizing amplifies the mass transfer in soil continuum. At initial soil stage the mass transfer in soil continuum plays positive role. Adverse substances are leached, in particular soluble salts. Fine material and organic particles determining soil fertility are accumulating. However, after a soil genesis initial stage the mass transfer through soil continuum plays negative role. Irrigation excess water flow into soil reduces the productivity of cultivated plants as compared to the optimum soil solution conditions. The excess soil moisture leads to excess transpiration, evaporation, infiltration, destroys the soil disperse system composition, forms inactive dead-end pores, leaches useful biological and other substances synthesized in soil out from active biosphere stage to vadose zone. These substances are entering the undesired stage of sedimentation and lithogenesis. Such adverse events are enhanced by irrigation. As a result, up to 80-90% of the fresh water taken for irrigation from lakes, rivers, storage reservoirs, desalinators are lost useless entailing economic losses. As a result of irrigation the quality of water is deteriorated as well as the quality of soil and landscape. A quality of human environment and a quality of biosphere as a whole is reduced. It is much more dangerous than economic losses. The irrigation paradigm shift is essential for successful water policy and water management in modern world. In a framework of Biogeosystem technics the new intrasoil pulse continuous-discrete paradigm of irrigation is developed. Water is supplied by small discrete portions into individual volumes of a soil continuum without excess soil mass transfer, transpiration, evaporation and seepage. New paradigm of irrigation optimizes plant growth, reduces consumption of water per unit of biological product, the yield increases. It provides the soil and landscape conservation, fresh water - the global deficit - saving up to 10-20 times, biological productivity and sustainability of biosphere. Intrasoil pulse continuous-discrete robotic irrigation technologies match the nowadays noosphere technological platform.

Probing Metabolic Activity of Deep Subseafloor Life with NanoSIMS

NASA Astrophysics Data System (ADS)

Morono, Y.; Terada, T.; Itoh, M.; Inagaki, F.

2014-12-01

There are very few natural environments where life is absent in the Earth's surface biosphere. However, uninhabitable region is expected to be exist in the deep subsurface biosphere, of which extent and constraining factor(s) have still remained largly unknown. Scientific ocean drilling have revealed that microbial communities in sediments are generally phylogenetically distinct from known spieces isolated from the Earth's surface biosphere, and hence metabolic functions of the deep subseafloor life remain unknown. In addition, activity of subseafloor microbial cells are thought to be extraordinally slow, as indicated by limited supply of neutrient and energy substrates. To understand the limits of the Earth's subseafloor biosphere and metabolic functions of microbial populations, detection and quantification of the deeply buried microbial cells in geological habitats are fundamentary important. Using newly developed cell separation techniques as well as an discriminative cell detection system, the current quantification limit of sedimentary microbial cells approaches to 102 cells/cm3. These techniques allow not only to assess very small microbial population close to the subsurface biotic fringe, but also to separate and sort the target cells using flow cytometric cell sorter. Once the deep subseafloor microbial cells are detached from mineral grains and sorted, it opens new windows to subsequent molecular ecological and element/isotopic analyses. With a combined use of nano-scale secondary ion masspectrometry (NanoSIMS) and stable isotope-probing techniques, it is possible to detect and measure activity of substrate incorporation into biomass, even for extremely slow metabolic processes such as uncharacteriszed deep subseafloor life. For example, it was evidenced by NanoSIMS that at least over 80% of microbial cells at ~200 meters-deep, 460,000-year-old sedimentary habitat are indeed live, which substrate incooporation was found to be low (10-15 gC/cell/day) even under the lab incubation condition. Also microbial activity in ultraoligotrophic biosphere samples such as the South Pacific Gyre (i.e., IODP Expeditions 329) will be shown. Our results demonstrates metabolic potential of microbes that have been survived for geological timescale in extremely starved condition.

Nitrogen Fertilization of Corn: Plant Biochemistry Effects and Carbon Cycle Implications

NASA Astrophysics Data System (ADS)

Gallagher, M. E.; Hockaday, W. C.; Masiello, C. A.; McSwiney, C. P.; Robertson, G. P.; Baldock, J. A.

2008-05-01

Atmospheric carbon dioxide (CO2) concentrations are rising due to anthropogenic CO2 emissions (Alley et al. 2007; Prentice et al. 2001). About half of the anthropogenic CO2 emitted during the 1990s was absorbed by the terrestrial biosphere and ocean (Prentice et al. 2001). It is possible to estimate the size of terrestrial and oceanic carbon sinks individually using atmospheric CO2 and O2 measurements (Keeling et al. 1996). To best estimate the sizes of these carbon sinks, we need to accurately know the oxidative ratio (OR) of the terrestrial biosphere (Randerson et al. 2006). OR is the ratio of the moles of O2 released per moles of CO2 consumed in gas fluxes between the terrestrial biosphere and atmosphere. Though it is likely that the net OR of the biosphere varies with ecosystem type and nutrient status, OR is assumed constant in carbon sink apportionment calculations (e.g. Prentice et al. 2001). Small shifts in OR can lead to large variations in the calculated sizes of the terrestrial biosphere and ocean carbon sinks (Randerson et al. 2006). OR likely shifts with changes in climate, nutrient status, and land use. These shifts are due, in part, to shifts in plant biochemistry. We are measuring ecosystem OR in corn agricultural ecosystems under a range of nitrogen fertilization treatments at the Kellogg Biological Station-Long Term Ecological Research Site (KBS-LTER) in Michigan. We measure OR indirectly, through its relationship with organic carbon oxidation state (Cox) (Masiello et al. in press 2008). Cox can be measured through elemental analysis and, with basic knowledge of plant nitrogen use patterns, Cox values can be converted to OR values. Cox can also be measured through 13C nuclear magnetic resonance spectroscopy (NMR), which can be combined with a molecular mixing model to determine Cox, OR, and plant biochemical composition (i.e. percentage carbohydrates, lignin, lipids, and proteins) (Baldock et al. 2004). Here we present data showing the effects of varying corn ecosystem nitrogen fertilization rates (from 0 to 292 kg N/ha) on ecosystem OR and plant biochemistry.

The Potential Impacts of Nutrient and CO2 Variations on Ecosystem Oxidative Ratio

NASA Astrophysics Data System (ADS)

Gallagher, M. E.; Hockaday, W. C.; Masiello, C. A.; Snapp, S.; Polley, W.; McSwiney, C. P.; Baldock, J.

2009-12-01

A fraction of fossil fuel carbon dioxide (CO2) emissions are being taken up by the terrestrial biosphere and the oceans. One particularly effective way of determining the sizes of these terrestrial biosphere and ocean carbon sinks is based on the measurements of changes in atmospheric oxygen (O2) and CO2 concentrations (Keeling et al. 1996). This method of carbon apportionment requires knowledge of total fossil fuel CO2 emissions, atmospheric O2 and CO2 concentrations, and the value of the terrestrial biosphere oxidative ratio (OR), which has historically been assumed to be constant at 1.10 (e.g. Prentice et al. 2001). OR is the ratio of moles of O2 per mole of CO2 in gas exchanges between the terrestrial biosphere and the atmosphere. An incorrect estimation of the biosphere’s OR results in misapportionment of CO2 between the terrestrial biosphere and ocean carbon sinks (Randerson et al. 2006). Understanding how OR can vary with changing environmental properties is therefore essential to accurately estimate the size of the terrestrial carbon sink. We estimate OR through its relationship with organic carbon oxidation state (Cox) measurements made using a 13C nuclear magnetic resonance spectrometer and a CHNSO elemental analyzer (Masiello et al. 2008; Hockaday et al. 2009). It is clear that ecosystem OR values frequently deviate from the assumed 1.10 (Masiello et al., 2008; Hockaday et al., 2009). Here we review what mechanisms drive shifts in OR, including: fire, climate (precipitation and temperature), land use change, atmospheric CO2 concentrations, and nutrient supply. We present data on the impact of nitrogen supply and elevated CO2 on ecosystem OR at two different field sites. We measure the effect of nitrogen supply on an agricultural ecosystem at the Kellogg Biological Station-Living Field Laboratory (KBS-LFL) in Michigan over a fertilization gradient (0 to 202 kg N/ha). We also measured the effect of atmospheric CO2 variation on ecosystem OR at a grassland site experiencing three atmospheric CO2 levels: pre-industrial, current, and projected (the USDA-Agricultural Research Service field site in Temple, Texas).

Agricultural conversion reduces biospheric vegetation productivity in the absence of external inputs

NASA Astrophysics Data System (ADS)

Smith, W. K.; Cleveland, C. C.; Reed, S.; Running, S. W.

2013-12-01

Increasing global population, energy demand, and standard of living has driven humanity to co-opt a growing share of the planet's natural resources resulting in many well-known environmental trade-offs. Here, we explored the impact of global-scale agricultural production on a basic resource fundamental to life on Earth: global terrestrial vegetation growth (net primary production; NPP). First, we compared current rates of agricultural NPP - derived from crop-specific agricultural statistics - with rates of natural NPP - derived from satellite measurements. Next, we disaggregated our results by climate zone, conversion type, crop type, management intensity, and region to identify where agricultural conversion has driven significant degradation of biospheric NPP. At the global-scale, our data indicate that agricultural conversion has resulted in a ~7% reduction in biospheric NPP (ΔNPP), although the impact varied widely at the pixel level. Positive ΔNPP values, signifying an increase in NPP due to agricultural conversion, occurred only in areas receiving significant external water and nutrient inputs (i.e., intensively managed areas). Conversely, negative ΔNPP values, signifying a reduction in NPP due to agricultural conversion, occurred over ~90% of agricultural lands globally, with the largest reductions in areas formerly occupied by tropical forests and savannas (71% and 66% reductions in NPP, respectively). Without new global-scale policies that explicitly consider changes in NPP due to land cover conversion, future demand-driven increases in agricultural output - likely dependent on some level of expansion into natural ecosystems - could continue to drive net declines in biospheric NPP, with potential detrimental consequences for global carbon storage. A spatially explicit estimate of the effect of agricultural land cover conversion on natural primary production for 20 staple crops. ΔNPP was estimated independently for a) irrigated, b) high input, c) low input, and d) subsistence management intensities. All remaining vegetated land is represented in grey, while barren land is represented in white. Globally, agricultural land cover conversion has reduced natural primary production by 3.0 × 0.68 Pg C y-1 (i.e., a ~7% reduction in biospheric NPP), with a disproportionately large percentage of this reduction attributable to the conversion of temperate (~44%) and tropical (~50%) ecosystems.

Achieving Closure for Bioregenerative Life Support Systems: Engineering and Ecological Challenges, Research Opportunities

NASA Astrophysics Data System (ADS)

Dempster, William; Allen, John P.

Closed systems are desirable for a number of purposes: space life support systems where precious life-supporting resources need to be kept inside; biospheric systems; where global ecological pro-cesses can be studied in great detail and testbeds where research topics requiring isolation from the outside (e.g. genetically modified organisms; radioisotopes) can be studied in isolation from the outside environment and where their ecological interactions and fluxes can be studied. But to achieve and maintain closure raises both engineering and ecological challenges. Engineering challenges include methods of achieving closure for structures of different materials, and devel-oping methods of allowing energy (for heating and cooling) and information transfer through the materially closed structure. Methods of calculating degree of closure include measuring degradation rates of inert trace gases introduced into the system. An allied problem is devel-oping means of locating where leaks are located so that they may be repaired and degree of closure maintained. Once closure is achieved, methods of dealing with the pressure differen-tials between inside and outside are needed: from inflatable structures which might adjust to the pressure difference to variable volume chambers attached to the life systems component. These issues are illustrated through the engineering employed at Biosphere 2, the Biosphere 2 Test Module and the Laboratory Biosphere and a discussion of methods used by other closed ecological system facility engineers. Ecological challenges include being able to handle faster cycling rates and accentuated daily and seasonal fluxes of critical life elements such as carbon dioxide, oxygen, water, macro-and mico-nutrients. The problems of achieving sustainability in closed systems for life support include how to handle atmospheric dynamics including trace gases, producing a complete human diet and recycling nutrients and maintaining soil fertility, healthy air and water and preventing the loss of crucial elements from active circulation. In biospheric facilities the challenge is also to produce analogue to natural biomes and ecosys-tems, studying processes of self-organization and adaptation in systems that allow specification or determination of state variables and cycles which may be followed through all interactions from atmosphere to soils.

False Negatives for Remote Life Detection on Ocean-Bearing Planets: Lessons from the Early Earth

NASA Astrophysics Data System (ADS)

Reinhard, Christopher T.; Olson, Stephanie L.; Schwieterman, Edward W.; Lyons, Timothy W.

2017-04-01

Ocean-atmosphere chemistry on Earth has undergone dramatic evolutionary changes throughout its long history, with potentially significant ramifications for the emergence and long-term stability of atmospheric biosignatures. Though a great deal of work has centered on refining our understanding of false positives for remote life detection, much less attention has been paid to the possibility of false negatives, that is, cryptic biospheres that are widespread and active on a planet's surface but are ultimately undetectable or difficult to detect in the composition of a planet's atmosphere. Here, we summarize recent developments from geochemical proxy records and Earth system models that provide insight into the long-term evolution of the most readily detectable potential biosignature gases on Earth - oxygen (O2), ozone (O3), and methane (CH4). We suggest that the canonical O2-CH4 disequilibrium biosignature would perhaps have been challenging to detect remotely during Earth's ˜4.5-billion-year history and that in general atmospheric O2/O3 levels have been a poor proxy for the presence of Earth's biosphere for all but the last ˜500 million years. We further suggest that detecting atmospheric CH4 would have been problematic for most of the last ˜2.5 billion years of Earth's history. More broadly, we stress that internal oceanic recycling of biosignature gases will often render surface biospheres on ocean-bearing silicate worlds cryptic, with the implication that the planets most conducive to the development and maintenance of a pervasive biosphere will often be challenging to characterize via conventional atmospheric biosignatures.

Carbon isotope discrimination of arctic and boreal biomes inferred from remote atmospheric measurements and a biosphere-atmosphere model

NASA Astrophysics Data System (ADS)

Randerson, J. T.; Still, C. J.; Ballé, J. J.; Fung, I. Y.; Doney, S. C.; Tans, P. P.; Conway, T. J.; White, J. W. C.; Vaughn, B.; Suits, N.; Denning, A. S.

2002-07-01

Estimating discrimination against 13C during photosynthesis at landscape, regional, and biome scales is difficult because of large-scale variability in plant stress, vegetation composition, and photosynthetic pathway. Here we present estimates of 13C discrimination for northern biomes based on a biosphere-atmosphere model and on National Oceanic and Atmospheric Administration Climate Monitoring and Diagnostics Laboratory and Institute of Arctic and Alpine Research remote flask measurements. With our inversion approach, we solved for three ecophysiological parameters of the northern biosphere (13C discrimination, a net primary production light use efficiency, and a temperature sensitivity of heterotrophic respiration (a Q10 factor)) that provided a best fit between modeled and observed δ13C and CO2. In our analysis we attempted to explicitly correct for fossil fuel emissions, remote C4 ecosystem fluxes, ocean exchange, and isotopic disequilibria of terrestrial heterotrophic respiration caused by the Suess effect. We obtained a photosynthetic discrimination for arctic and boreal biomes between 19.0 and 19.6‰. Our inversion analysis suggests that Q10 and light use efficiency values that minimize the cost function covary. The optimal light use efficiency was 0.47 gC MJ-1 photosynthetically active radiation, and the optimal Q10 value was 1.52. Fossil fuel and ocean exchange contributed proportionally more to month-to-month changes in the atmospheric growth rate of δ13C and CO2 during winter months, suggesting that remote atmospheric observations during the summer may yield more precise estimates of the isotopic composition of the biosphere.

Engineering Challenges for Closed Ecological System facilities

NASA Astrophysics Data System (ADS)

Dempster, William; Nelson, Mark; Allen, John P.

2012-07-01

Engineering challenges for closed ecological systems include methods of achieving closure for structures of different materials, and developing methods of allowing energy (for heating and cooling) and information transfer through the materially closed structure. Methods of calculating degree of closure include measuring degradation rates of inert trace gases introduced into the system. An allied problem is developing means of locating where leaks are located so that they may be repaired and degree of closure maintained. Once closure is achieved, methods of dealing with the pressure differentials between inside and outside are needed: from inflatable structures which might adjust to the pressure difference to variable volume chambers attached to the life systems component. These issues are illustrated through the engineering employed at Biosphere 2, the Biosphere 2 Test Module and the Laboratory Biosphere and a discussion of methods used by other closed ecological system facility engineers. Ecological challenges include being able to handle faster cycling rates and accentuated daily and seasonal fluxes of critical life elements such as carbon dioxide, oxygen, water, macro- and mico-nutrients. The problems of achieving sustainability in closed systems for life support include how to handle atmospheric dynamics including trace gases, producing a complete human diet and recycling nutrients and maintaining soil fertility, healthy air and water and preventing the loss of crucial elements from active circulation. In biospheric facilities the challenge is also to produce analogue to natural biomes and ecosystems, studying processes of self-organization and adaptation in systems that allow specification or determination of state variables and cycles which may be followed through all interactions from atmosphere to soils.

Problem-Solving Test: Pyrosequencing

ERIC Educational Resources Information Center

Szeberenyi, Jozsef

2013-01-01

Terms to be familiar with before you start to solve the test: Maxam-Gilbert sequencing, Sanger sequencing, gel electrophoresis, DNA synthesis reaction, polymerase chain reaction, template, primer, DNA polymerase, deoxyribonucleoside triphosphates, orthophosphate, pyrophosphate, nucleoside monophosphates, luminescence, acid anhydride bond,…

Fluorogenic DNA Sequencing in PDMS Microreactors

PubMed Central

Sims, Peter A.; Greenleaf, William J.; Duan, Haifeng; Xie, X. Sunney

2012-01-01

We have developed a multiplex sequencing-by-synthesis method combining terminal-phosphate labeled fluorogenic nucleotides (TPLFNs) and resealable microreactors. In the presence of phosphatase, the incorporation of a non-fluorescent TPLFN into a DNA primer by DNA polymerase results in a fluorophore. We immobilize DNA templates within polydimethylsiloxane (PDMS) microreactors, sequentially introduce one of the four identically labeled TPLFNs, seal the microreactors, allow template-directed TPLFN incorporation, and measure the signal from the fluorophores trapped in the microreactors. This workflow allows sequencing in a manner akin to pyrosequencing but without constant monitoring of each microreactor. With cycle times of <10 minutes, we demonstrate 30 base reads with ∼99% raw accuracy. “Fluorogenic pyrosequencing” combines benefits of pyrosequencing, such as rapid turn-around, native DNA generation, and single-color detection, with benefits of fluorescence-based approaches, such as highly sensitive detection and simple parallelization. PMID:21666670

Multiple approaches to characterize the microbial community in a thermophilic anaerobic digester running on swine manure: a case study.

PubMed

Tuan, Nguyen Ngoc; Chang, Yi-Chia; Yu, Chang-Ping; Huang, Shir-Ly

2014-01-01

In this study, the first survey of microbial community in thermophilic anaerobic digester using swine manure as sole feedstock was performed by multiple approaches including denaturing gradient gel electrophoresis (DGGE), clone library and pyrosequencing techniques. The integrated analysis of 21 DGGE bands, 126 clones and 8506 pyrosequencing read sequences revealed that Clostridia from the phylum Firmicutes account for the most dominant Bacteria. In addition, our analysis also identified additional taxa that were missed by the previous researches, including members of the bacterial phyla Synergistetes, Planctomycetes, Armatimonadetes, Chloroflexi and Nitrospira which might also play a role in thermophilic anaerobic digester. Most archaeal 16S rRNA sequences could be assigned to the order Methanobacteriales instead of Methanomicrobiales comparing to previous studies. In addition, this study reported that the member of Methanothermobacter genus was firstly found in thermophilic anaerobic digester. Copyright © 2014 Elsevier GmbH. All rights reserved.

Novel multiplex qualitative detection using universal primer-multiplex-PCR combined with pyrosequencing.

PubMed

Shang, Ying; Xu, Wentao; Wang, Yong; Xu, Yuancong; Huang, Kunlun

2017-12-15

This study described a novel multiplex qualitative detection method using pyrosequencing. Based on the principle of the universal primer-multiplex-PCR, only one sequencing primer was employed to realize the detection of the multiple targets. Samples containing three genetically modified (GM) crops in different proportions were used to validate the method. The dNTP dispensing order was designed based on the product sequences. Only 12 rounds (ATCTGATCGACT) of dNTPs addition and, often, as few as three rounds (CAT) under ideal conditions, were required to detect the GM events qualitatively, and sensitivity was as low as 1% of a mixture. However, when considering a mixture, calculating signal values allowed the proportion of each GM to be estimated. Based on these results, we concluded that our novel method not only realized detection but also allowed semi-quantitative detection of individual events. Copyright © 2017. Published by Elsevier Ltd.

Evaluation of feed grade sodium bisulfate impact on gastrointestinal tract microbiota ecology in broilers via a pyrosequencing platform.

PubMed

Park, Si Hong; Dowd, Scot E; McReynolds, Jack L; Byrd, James A; Nisbet, David J; Ricke, Steven C

2015-12-01

The gastrointestinal microbial community in broiler chickens consists of many different species of bacteria, and the overall microbiota can vary from bird to bird. To control pathogenic bacteria in broilers and improve gut health, numerous potential dietary amendments have been used. In this study, we used a pyrosequencing platform to evaluate the effect of sodium bisulfate on microbiota of the crop, cecum, and ileum of broiler chickens grown over several weeks. The diversity information in each digestive organ sample exhibited considerable variation and was clearly separable, suggesting distinct bacterial populations. Although no apparent microbial clustering occurred between the control and the dietary treatments, we did observe shifts in overall microbiota populations in the crop, ileum, and ceca as well as changes in specific microorganisms such as Bacteroides, Clostridium, and Lactobacillus species that were identified as birds became older. © 2015 Poultry Science Association Inc.

Performances of two biotrickling filters in treating H₂S-containing waste gases and analysis of corresponding bacterial communities by pyrosequencing.

PubMed

Li, Jianjun; Ye, Guangyun; Sun, Duanfang; Sun, Guoping; Zeng, Xiaowei; Xu, Jian; Liang, Shizhong

2012-09-01

Two identical biotrickling filters named BTFa and BTFb were run in parallel to examine their performances in removing hydrogen sulfide. BTFa was filled with ceramic granules, and BTFb was filled with volcanic rocks. The results showed that BTFb was more robust than BTFa under acidic conditions. At empty bed residence times (EBRTs) of 20 and 15 s, the removal efficiency of BTFa was close to 100%. At EBRTs of 10 and 5 s, the removal efficiency of BTFa slightly decreased. The removal efficiencies of BTFa decreased by different degrees at the end of each stage, dropping to 94%, 81%, 60%, and 71%, respectively. However, the H(2)S removal efficiency in BTFb consistently reached 99% throughout the experiment. Pyrosequencing analyses indicated that members of Thiomonas dominated in both BTFs, but the relative abundance of Acidithiobacillus was higher in BTFb than in BTFa.

Forensic discrimination of vaginal epithelia by DNA methylation analysis through pyrosequencing.

PubMed

Antunes, Joana; Silva, Deborah S B S; Balamurugan, Kuppareddi; Duncan, George; Alho, Clarice S; McCord, Bruce

2016-10-01

The accurate identification of body fluids from crime scenes can aid in the discrimination between criminal and innocent intent. This research aimed to determine if the levels of DNA methylation in the locus PFN3A could be used to discriminate vaginal epithelia from other body fluids. In this work we bisulfite-modified and amplified DNA samples from blood, saliva, semen, and vaginal epithelia using primers for PFN3A. Through pyrosequencing we were able to show that vaginal epithelia present distinct methylation levels when compared to other body fluids. Mixtures of different body fluids present methylation values that correlate with single-source body fluid samples and the primers for PFN3A are specific for primates. This report successfully demonstrated that the analysis of methylation in the PFN3A locus can be used for vaginal epithelia discrimination in forensic samples. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Cosmic Rays as a Factor of Biospheric Evolution

NASA Astrophysics Data System (ADS)

Miroshnichenko, L. I.

2014-10-01

There are no doubts that the Earth's space environment in the past inevitably has exerted direct and/or indirect influence [1-4] on the conditions of terrestrial life and biospheric evolution. Well-known space factors are usually the fluxes of cosmic dust and gas, comets and asteroids, cosmic rays (energetic particles of galactic and/or solar origin), interplanetary plasma (solar wind) and electromagnetic radiation of different energies, wave lengths, or frequencies. Of great interest are radiation conditions and their variations, especially in the remote past (over the geological time scales). The Sun, the most important and indispensable condition for the existence of the Earth's biosphere, is also a potential source of dangerous emissions. In continuation of (and in addition to) our review paper [3], below we summarize the observational data and results of theoretical works that have been carried out and/or published mainly after 2012. These studies are actually in the frontier region between the Astrobiology and Space Weather. Our main attention is paid to cosmic rays (CR) of galactic and solar origin (GCR and SCR, respectively).

Maximum entropy production in environmental and ecological systems.

PubMed

Kleidon, Axel; Malhi, Yadvinder; Cox, Peter M

2010-05-12

The coupled biosphere-atmosphere system entails a vast range of processes at different scales, from ecosystem exchange fluxes of energy, water and carbon to the processes that drive global biogeochemical cycles, atmospheric composition and, ultimately, the planetary energy balance. These processes are generally complex with numerous interactions and feedbacks, and they are irreversible in their nature, thereby producing entropy. The proposed principle of maximum entropy production (MEP), based on statistical mechanics and information theory, states that thermodynamic processes far from thermodynamic equilibrium will adapt to steady states at which they dissipate energy and produce entropy at the maximum possible rate. This issue focuses on the latest development of applications of MEP to the biosphere-atmosphere system including aspects of the atmospheric circulation, the role of clouds, hydrology, vegetation effects, ecosystem exchange of energy and mass, biogeochemical interactions and the Gaia hypothesis. The examples shown in this special issue demonstrate the potential of MEP to contribute to improved understanding and modelling of the biosphere and the wider Earth system, and also explore limitations and constraints to the application of the MEP principle.

Long term changes in forest cover and land use of Similipal Biosphere Reserve of India using satellite remote sensing data

NASA Astrophysics Data System (ADS)

Saranya, K. R. L.; Reddy, C. Sudhakar

2016-04-01

The spatial changes in forest cover of Similipal biosphere reserve, Odisha, India over eight decades (1930-2012) has been quantified by using multi-temporal data from different sources. Over the period, the forest cover reduced by 970.8 km2 (23.6% of the total forest), and most significantly during the period, 1930-1975. Human-induced activities like conversion of forest land for agriculture, construction of dams and mining activities have been identified as major drivers of deforestation. Spatial analysis indicates that 399 grids (1 grid = 1 × 1 km) have undergone large-scale changes in forest cover (>75 ha) during 1930-1975, while only 3 grids have shown >75 ha loss during 1975-1990. Annual net rate of deforestation was 0.58 during 1930-1975, which has been reduced substantially during 1975-1990 (0.04). Annual gross rate of deforestation in 2006-2012 is indeed low (0.01) as compared to the national and global average. This study highlights the impact and effectiveness of conservation practices in minimizing the rate of deforestation and protecting the Similipal Biosphere Reserve.

Water: the bloodstream of the biosphere.

PubMed

Ripl, Wilhelm

2003-12-29

Water, the bloodstream of the biosphere, determines the sustainability of living systems. The essential role of water is expanded in a conceptual model of energy dissipation, based on the water balance of whole landscapes. In this model, the underlying role of water phase changes--and their energy-dissipative properties--in the function and the self-organized development of natural systems is explicitly recognized. The energy-dissipating processes regulate the ecological dynamics within the Earth's biosphere, in such a way that the development of natural systems is never allowed to proceed in an undirected or random way. A fundamental characteristic of self-organized development in natural systems is the increasing role of cyclic processes while loss processes are correspondingly reduced. This gives a coincidental increase in system efficiency, which is the basis of growing stability and sustainability. Growing sustainability can be seen as an increase of ecological efficiency, which is applicable at all levels up to whole landscapes. Criteria for necessary changes in society and for the design of the measures that are necessary to restore sustainable landscapes and waters are derived.

Water: the bloodstream of the biosphere.

PubMed Central

Ripl, Wilhelm

2003-01-01

Water, the bloodstream of the biosphere, determines the sustainability of living systems. The essential role of water is expanded in a conceptual model of energy dissipation, based on the water balance of whole landscapes. In this model, the underlying role of water phase changes--and their energy-dissipative properties--in the function and the self-organized development of natural systems is explicitly recognized. The energy-dissipating processes regulate the ecological dynamics within the Earth's biosphere, in such a way that the development of natural systems is never allowed to proceed in an undirected or random way. A fundamental characteristic of self-organized development in natural systems is the increasing role of cyclic processes while loss processes are correspondingly reduced. This gives a coincidental increase in system efficiency, which is the basis of growing stability and sustainability. Growing sustainability can be seen as an increase of ecological efficiency, which is applicable at all levels up to whole landscapes. Criteria for necessary changes in society and for the design of the measures that are necessary to restore sustainable landscapes and waters are derived. PMID:14728789

Prospects for the Study of Evolution in the Deep Biosphere

PubMed Central

Biddle, Jennifer F.; Sylvan, Jason B.; Brazelton, William J.; Tully, Benjamin J.; Edwards, Katrina J.; Moyer, Craig L.; Heidelberg, John F.; Nelson, William C.

2012-01-01

Since the days of Darwin, scientists have used the framework of the theory of evolution to explore the interconnectedness of life on Earth and adaptation of organisms to the ever-changing environment. The advent of molecular biology has advanced and accelerated the study of evolution by allowing direct examination of the genetic material that ultimately determines the phenotypes upon which selection acts. The study of evolution has been furthered through examination of microbial evolution, with large population numbers, short generation times, and easily extractable DNA. Such work has spawned the study of microbial biogeography, with the realization that concepts developed in population genetics may be applicable to microbial genomes (Martiny et al., 2006; Manhes and Velicer, 2011). Microbial biogeography and adaptation has been examined in many different environments. Here we argue that the deep biosphere is a unique environment for the study of evolution and list specific factors that can be considered and where the studies may be performed. This publication is the result of the NSF-funded Center for Dark Energy Biosphere Investigations (C-DEBI) theme team on Evolution (www.darkenergybiosphere.org). PMID:22319515

Prospects for the study of evolution in the deep biosphere.

PubMed

Biddle, Jennifer F; Sylvan, Jason B; Brazelton, William J; Tully, Benjamin J; Edwards, Katrina J; Moyer, Craig L; Heidelberg, John F; Nelson, William C

2011-01-01

Since the days of Darwin, scientists have used the framework of the theory of evolution to explore the interconnectedness of life on Earth and adaptation of organisms to the ever-changing environment. The advent of molecular biology has advanced and accelerated the study of evolution by allowing direct examination of the genetic material that ultimately determines the phenotypes upon which selection acts. The study of evolution has been furthered through examination of microbial evolution, with large population numbers, short generation times, and easily extractable DNA. Such work has spawned the study of microbial biogeography, with the realization that concepts developed in population genetics may be applicable to microbial genomes (Martiny et al., 2006; Manhes and Velicer, 2011). Microbial biogeography and adaptation has been examined in many different environments. Here we argue that the deep biosphere is a unique environment for the study of evolution and list specific factors that can be considered and where the studies may be performed. This publication is the result of the NSF-funded Center for Dark Energy Biosphere Investigations (C-DEBI) theme team on Evolution (www.darkenergybiosphere.org).

Constraints on natural global atmospheric CO2 fluxes from 1860 to 2010 using a simplified explicit forward model

PubMed Central

Hellevang, Helge; Aagaard, Per

2015-01-01

Land-use changes until the beginning of the 20th century made the terrestrial biosphere a net source of atmospheric carbon. Later, burning of fossil fuel surpassed land use changes as the major anthropogenic source of carbon. The terrestrial biosphere is at present suggested to be a carbon sink, but the distribution of excess anthropogenic carbon to the ocean and biosphere sinks is highly uncertain. Our modeling suggest that land-use changes can be tracked quite well by the carbon isotopes until mid-20th century, whereas burning of fossil fuel dominates the present-day observed changes in the isotope signature. The modeling indicates that the global carbon isotope fractionation has not changed significantly during the last 150 years. Furthermore, increased uptake of carbon by the ocean and increasing temperatures does not yet appear to have resulted in increasing the global gross ocean-to-atmosphere carbon fluxes. This may however change in the future when the excess carbon will emerge in the ocean upwelling zones, possibly reducing the net-uptake of carbon compared to the present-day ocean. PMID:26611741

A socio-environmental monitoring system for a UNESCO biosphere reserve.

PubMed

Lowell, Kim

2017-11-03

To identify potentially critical changes in an area's ongoing ability to produce multiple ecosystem services, a monitoring system was designed and implemented for the Mornington Peninsula and Westernport Biosphere Reserve in southeast Australia. The system is underpinned by an "environmental vital signs" (EVS) approach that was adopted to provide early warning of critical changes in human and natural characteristics of the area. The six themes monitored are non-coastal water, land including vegetation, biodiversity, natural heritage, built environment (including human population and economic activity), and coasts. These are monitored for the entire area, and each of its five constituent town council areas. After a critical change in any of these is identified, further investigation is required to identify causal factors and, if required, determine an appropriate response. The system relies on data available from external (third-party) organisations to monitor the natural and human characteristics of the area that were important in its designation as a UNESCO biosphere reserve. Strengths and weaknesses associated with the use of third-party data are discussed. These include adoption of baseline years and data reporting periods for different factors, costs, and data quality.

The carbon cycle on early Earth--and on Mars?

PubMed

Grady, Monica M; Wright, Ian

2006-10-29

One of the goals of the present Martian exploration is to search for evidence of extinct (or even extant) life. This could be redefined as a search for carbon. The carbon cycle (or, more properly, cycles) on Earth is a complex interaction among three reservoirs: the atmosphere; the hydrosphere; and the lithosphere. Superimposed on this is the biosphere, and its presence influences the fixing and release of carbon in these reservoirs over different time-scales. The overall carbon balance is kept at equilibrium on the surface by a combination of tectonic processes (which bury carbon), volcanism (which releases it) and biology (which mediates it). In contrast to Earth, Mars presently has no active tectonic system; neither does it possess a significant biosphere. However, these observations might not necessarily have held in the past. By looking at how Earth's carbon cycles have changed with time, as both the Earth's tectonic structure and a more sophisticated biology have evolved, and also by constructing a carbon cycle for Mars based on the carbon chemistry of Martian meteorites, we investigate whether or not there is evidence for a Martian biosphere.

The carbon cycle on early Earth—and on Mars?

PubMed Central

Grady, Monica M; Wright, Ian

2006-01-01

One of the goals of the present Martian exploration is to search for evidence of extinct (or even extant) life. This could be redefined as a search for carbon. The carbon cycle (or, more properly, cycles) on Earth is a complex interaction among three reservoirs: the atmosphere; the hydrosphere; and the lithosphere. Superimposed on this is the biosphere, and its presence influences the fixing and release of carbon in these reservoirs over different time-scales. The overall carbon balance is kept at equilibrium on the surface by a combination of tectonic processes (which bury carbon), volcanism (which releases it) and biology (which mediates it). In contrast to Earth, Mars presently has no active tectonic system; neither does it possess a significant biosphere. However, these observations might not necessarily have held in the past. By looking at how Earth's carbon cycles have changed with time, as both the Earth's tectonic structure and a more sophisticated biology have evolved, and also by constructing a carbon cycle for Mars based on the carbon chemistry of Martian meteorites, we investigate whether or not there is evidence for a Martian biosphere. PMID:17008211

Imposing strong constraints on tropical terrestrial CO2 fluxes using passenger aircraft based measurements

NASA Astrophysics Data System (ADS)

Niwa, Yosuke; Machida, Toshinobu; Sawa, Yousuke; Matsueda, Hidekazu; Schuck, Tanja J.; Brenninkmeijer, Carl A. M.; Imasu, Ryoichi; Satoh, Masaki

2012-06-01

Because very few measurements of atmospheric carbon dioxide (CO2) are available in the tropics, estimates of surface CO2 fluxes in tropical regions are beset with considerable uncertainties. To improve estimates of tropical terrestrial fluxes, atmospheric CO2 inversion was performed using passenger aircraft based measurements of the Comprehensive Observation Network for Trace gases by Airliner (CONTRAIL) project in addition to the surface measurement data set of GLOBALVIEW-CO2. Regional monthly fluxes at the earth's surface were estimated using the Bayesian synthesis approach focusing on the period 2006-2008 using the Nonhydrostatic Icosahedral Atmospheric Model-based Transport Model (NICAM-TM). By adding the aircraft to the surface data, the posterior flux errors were greatly reduced; specifically, error reductions of up to 64% were found for tropical Asia regions. This strong impact is closely related to efficient vertical transport in the tropics. The optimized surface fluxes using the CONTRAIL data were evaluated by comparing the simulated atmospheric CO2 distributions with independent aircraft measurements of the Civil Aircraft for the Regular Investigation of the atmosphere Based on an Instrument Container (CARIBIC) project. The inversion with the CONTRAIL data yields the global carbon sequestration rates of 2.22 ± 0.28 Pg C yr-1 for the terrestrial biosphere and 2.24 ± 0.27 Pg C yr-1 for the oceans (the both are adjusted by riverine input of CO2). For the first time the CONTRAIL CO2 measurements were used in an inversion system to identify the areas of greatest impact in terms of reducing flux uncertainties.

Next-Generation Genotyping by Digital PCR to Detect and Quantify the BRAF V600E Mutation in Melanoma Biopsies.

PubMed

Lamy, Pierre-Jean; Castan, Florence; Lozano, Nicolas; Montélion, Cécile; Audran, Patricia; Bibeau, Frédéric; Roques, Sylvie; Montels, Frédéric; Laberenne, Anne-Claire

2015-07-01

The detection of the BRAF V600E mutation in melanoma samples is used to select patients who should respond to BRAF inhibitors. Different techniques are routinely used to determine BRAF status in clinical samples. However, low tumor cellularity and tumor heterogeneity can affect the sensitivity of somatic mutation detection. Digital PCR (dPCR) is a next-generation genotyping method that clonally amplifies nucleic acids and allows the detection and quantification of rare mutations. Our aim was to evaluate the clinical routine performance of a new dPCR-based test to detect and quantify BRAF mutation load in 47 paraffin-embedded cutaneous melanoma biopsies. We compared the results obtained by dPCR with high-resolution melting curve analysis and pyrosequencing or with one of the allele-specific PCR methods available on the market. dPCR showed the lowest limit of detection. dPCR and allele-specific amplification detected the highest number of mutated samples. For the BRAF mutation load quantification both dPCR and pyrosequencing gave similar results with strong disparities in allele frequencies in the 47 tumor samples under study (from 0.7% to 79% of BRAF V600E mutations/sample). In conclusion, the four methods showed a high degree of concordance. dPCR was the more-sensitive method to reliably and easily detect mutations. Both pyrosequencing and dPCR could quantify the mutation load in heterogeneous tumor samples. Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

The microbiota of marketed processed edible insects as revealed by high-throughput sequencing.

PubMed

Garofalo, Cristiana; Osimani, Andrea; Milanović, Vesna; Taccari, Manuela; Cardinali, Federica; Aquilanti, Lucia; Riolo, Paola; Ruschioni, Sara; Isidoro, Nunzio; Clementi, Francesca

2017-04-01

Entomophagy has been linked to nutritional, economic, social and ecological benefits. However, scientific studies on the potential safety risks in eating edible insects need to be carried out for legislators, markets and consumers. In this context, the microbiota of edible insects deserves to be deeply investigated. The aim of this study was to elucidate the microbial species occurring in some processed marketed edible insects, namely powdered small crickets, whole dried small crickets (Acheta domesticus), whole dried locusts (Locusta migratoria), and whole dried mealworm larvae (Tenebrio molitor), through culture-dependent (classical microbiological analyses) and -independent methods (pyrosequencing). A great bacterial diversity and variation among insects was seen. Relatively low counts of total mesophilic aerobes, Enterobacteriaceae, lactic acid bacteria, Clostridium perfringens spores, yeasts and moulds in all of the studied insect batches were found. Furthermore, the presence of several gut-associated bacteria, some of which may act as opportunistic pathogens in humans, were found through pyrosequencing. Food spoilage bacteria were also identified, as well as Spiroplasma spp. in mealworm larvae, which has been found to be related to neurodegenerative diseases in animals and humans. Although viable pathogens such as Salmonella spp. and Listeria monocytogenes were not detected, the presence of Listeria spp., Staphylococcus spp., Clostridium spp. and Bacillus spp. (with low abundance) was also found through pyrosequencing. The results of this study contribute to the elucidation of the microbiota associated with edible insects and encourage further studies aimed to evaluate the influence of rearing and processing conditions on that microbiota. Copyright © 2016 Elsevier Ltd. All rights reserved.

Use of pyrosequencing to characterize the microbiota in the ileum of goats fed with increasing proportion of dietary grain.

PubMed

Mao, Shengyong; Huo, Wenjie; Zhu, Weiyun

2013-09-01

This study evaluated the effects of an increasing proportion of dietary grain on changes in bacterial populations in the goat ileum. Nine ruminally fistulated, castrated male goats were assigned to three diets in a completely randomized design. Goats were fed three different dietary treatments containing different proportions of corn grain (0, 25, and 50 %). The pH of the ileal contents and rumen fluid (P = 0.015) linearly decreased (P < 0.001), and the acetate, propionate, butyrate, and total volatile fatty acid in ileal contents increased (P < 0.05) with increases in dietary corn, and similar results were also observed in rumen fluid. The barcoded DNA pyrosequencing method was used to reveal 8 phyla, 70 genera, and 1,693 16S operational taxonomic units (OTUs). At the genus level, the proportions of Acetitomaculum, Enterococcus, Atopobium, unclassified Coriobacteriaceae, and unclassified Planctomycetaceae were linearly decreased (P < 0.05) with increases in corn grain. At the species level, high grain feeding linearly decreased the percentage of OTU8686 (unclassified Bacteria) (P = 0.004). To the best of our knowledge, this is the first study using barcoded DNA pyrosequencing method to survey the ileal microbiome of goats and the results suggest that increasing levels of dietary corn change the composition of the ileal bacterial community. These findings provide previously unknown information about the ileal microbiota of goats and a new understanding of the ileal microbial ecology, which may be useful in modulating the gut microbiome.

A comprehensive survey of soil acidobacterial diversity using pyrosequencing and clone library analyses

PubMed Central

Jones, Ryan T; Robeson, Michael S; Lauber, Christian L; Hamady, Micah; Knight, Rob; Fierer, Noah

2010-01-01

Acidobacteria are ubiquitous and abundant members of soil bacterial communities. However, an ecological understanding of this important phylum has remained elusive because its members have been difficult to culture and few molecular investigations have focused exclusively on this group. We generated an unprecedented number of acidobacterial DNA sequence data using pyrosequencing and clone libraries (39 707 and 1787 sequences, respectively) to characterize the relative abundance, diversity and composition of acidobacterial communities across a range of soil types. To gain insight into the ecological characteristics of acidobacterial taxa, we investigated the large-scale biogeographic patterns exhibited by acidobacterial communities, and related soil and site characteristics to acidobacterial community assemblage patterns. The 87 soils analyzed by pyrosequencing contained more than 8600 unique acidobacterial phylotypes (at the 97% sequence similarity level). One phylotype belonging to Acidobacteria subgroup 1, but not closely related to any cultured representatives, was particularly abundant, accounting for 7.4% of bacterial sequences and 17.6% of acidobacterial sequences, on average, across the soils. The abundance of Acidobacteria relative to other bacterial taxa was highly variable across the soils examined, but correlated strongly with soil pH (R = −0.80, P<0.001). Soil pH was also the best predictor of acidobacterial community composition, regardless of how the communities were characterized, and the relative abundances of the dominant Acidobacteria subgroups were readily predictable. Acidobacterial communities were more phylogenetically clustered as soil pH departed from neutrality, suggesting that pH is an effective habitat filter, restricting community membership to progressively more narrowly defined lineages as pH deviates from neutrality. PMID:19129864

Analysis of bacterial xylose isomerase gene diversity using gene-targeted metagenomics.

PubMed

Nurdiani, Dini; Ito, Michihiro; Maruyama, Toru; Terahara, Takeshi; Mori, Tetsushi; Ugawa, Shin; Takeyama, Haruko

2015-08-01

Bacterial xylose isomerases (XI) are promising resources for efficient biofuel production from xylose in lignocellulosic biomass. Here, we investigated xylose isomerase gene (xylA) diversity in three soil metagenomes differing in plant vegetation and geographical location, using an amplicon pyrosequencing approach and two newly-designed primer sets. A total of 158,555 reads from three metagenomic DNA replicates for each soil sample were classified into 1127 phylotypes, detected in triplicate and defined by 90% amino acid identity. The phylotype coverage was estimated to be within the range of 84.0-92.7%. The xylA gene phylotypes obtained were phylogenetically distributed across the two known xylA groups. They shared 49-100% identities with their closest-related XI sequences in GenBank. Phylotypes demonstrating <90% identity with known XIs in the database accounted for 89% of the total xylA phylotypes. The differences among xylA members and compositions within each soil sample were significantly smaller than they were between different soils based on a UniFrac distance analysis, suggesting soil-specific xylA genotypes and taxonomic compositions. The differences among xylA members and their compositions in the soil were strongly correlated with 16S rRNA variation between soil samples, also assessed by amplicon pyrosequencing. This is the first report of xylA diversity in environmental samples assessed by amplicon pyrosequencing. Our data provide information regarding xylA diversity in nature, and can be a basis for the screening of novel xylA genotypes for practical applications. Copyright © 2015. Published by Elsevier B.V.

Maternal blood contamination of collected cord blood can be identified using DNA methylation at three CpGs.

PubMed

Morin, Alexander M; Gatev, Evan; McEwen, Lisa M; MacIsaac, Julia L; Lin, David T S; Koen, Nastassja; Czamara, Darina; Räikkönen, Katri; Zar, Heather J; Koenen, Karestan; Stein, Dan J; Kobor, Michael S; Jones, Meaghan J

2017-01-01

Cord blood is a commonly used tissue in environmental, genetic, and epigenetic population studies due to its ready availability and potential to inform on a sensitive period of human development. However, the introduction of maternal blood during labor or cross-contamination during sample collection may complicate downstream analyses. After discovering maternal contamination of cord blood in a cohort study of 150 neonates using Illumina 450K DNA methylation (DNAm) data, we used a combination of linear regression and random forest machine learning to create a DNAm-based screening method. We identified a panel of DNAm sites that could discriminate between contaminated and non-contaminated samples, then designed pyrosequencing assays to pre-screen DNA prior to being assayed on an array. Maternal contamination of cord blood was initially identified by unusual X chromosome DNA methylation patterns in 17 males. We utilized our DNAm panel to detect contaminated male samples and a proportional amount of female samples in the same cohort. We validated our DNAm screening method on an additional 189 sample cohort using both pyrosequencing and DNAm arrays, as well as 9 publically available cord blood 450K data sets. The rate of contamination varied from 0 to 10% within these studies, likely related to collection specific methods. Maternal blood can contaminate cord blood during sample collection at appreciable levels across multiple studies. We have identified a panel of markers that can be used to identify this contamination, either post hoc after DNAm arrays have been completed, or in advance using a targeted technique like pyrosequencing.