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Purifying, Separating, and Concentrating Cells From a Sample Low in Biomass

Frequently there is an inability to process and analyze samples of low biomass due to limiting amounts of relevant biomaterial in the sample. Furthermore, molecular biological protocols geared towards increasing the density of recovered cells and biomolecules of interest, by their very nature, also concentrate unwanted inhibitory humic acids and other particulates that have an adversarial effect on downstream analysis. A novel and robust fluorescence-activated cell-sorting (FACS)-based technology has been developed for purifying (removing cells from sampling matrices), separating (based on size, density, morphology), and concentrating cells (spores, prokaryotic, eukaryotic) from a sample low in biomass. The technology capitalizes on fluorescent cell-sorting technologies to purify and concentrate bacterial cells from a low-biomass, high-volume sample. Over the past decade, cell-sorting detection systems have undergone enhancements and increased sensitivity, making bacterial cell sorting a feasible concept. Although there are many unknown limitations with regard to the applicability of this technology to environmental samples (smaller cells, few cells, mixed populations), dogmatic principles support the theoretical effectiveness of this technique upon thorough testing and proper optimization. Furthermore, the pilot study from which this report is based proved effective and demonstrated this technology capable of sorting and concentrating bacterial endospore and bacterial cells of varying size and morphology. Two commercial off-the-shelf bacterial counting kits were used to optimize a bacterial stain/dye FACS protocol. A LIVE/DEAD BacLight Viability and Counting Kit was used to distinguish between the live and dead cells. A Bacterial Counting Kit comprising SYTO BC (mixture of SYTO dyes) was employed as a broad-spectrum bacterial counting agent. Optimization using epifluorescence microscopy was performed with these two dye/stains. This refined protocol was further validated using varying ratios and mixtures of cells to ensure homogenous staining compared to that of individual cells, and were utilized for flow analyzer and FACS labeling. This technology focuses on the purification and concentration of cells from low-biomass spacecraft assembly facility samples. Currently, purification and concentration of low-biomass samples plague planetary protection downstream analyses. Having a capability to use flow cytometry to concentrate cells out of low-biomass, high-volume spacecraft/ facility sample extracts will be of extreme benefit to the fields of planetary protection and astrobiology. Successful research and development of this novel methodology will significantly increase the knowledge base for designing more effective cleaning protocols, and ultimately lead to a more empirical and true account of the microbial diversity present on spacecraft surfaces. Refined cleaning and an enhanced ability to resolve microbial diversity may decrease the overall cost of spacecraft assembly and/or provide a means to begin to assess challenging planetary protection missions.

Benardini, James N.↗

GEDI Launches a New Era of Biomass Inference from Space

Accurate estimation of aboveground forest biomass stocks is required to assess the impacts of land use changes such as deforestation and subsequent regrowth on concentrations of atmospheric CO 2 . The Global Ecosystem Dynamics Investigation (GEDI) is a lidar mission launched by NASA to the International Space Station in 2018. GEDI was specifically designed to retrieve vegetation structure within a novel, theoretical sampling design that explicitly quantifies biomass and its uncertainty across a variety of spatial scales. In this paper we provide the estimates of pan-tropical and temperate biomass derived from two years of GEDI observations. We present estimates of mean biomass densities at 1 km resolution, as well as estimates aggregated to the national level for every country GEDI observes, and at the sub-national level for the United States. For all estimates we provide the standard error of the mean biomass. These data serve as a baseline for current biomass stocks and their future changes, and the mission’s integrated use of formal statistical inference points the way towards the possibility of a new generation of powerful monitoring tools from space.

Ralph Dubayah↗

NASA's Biomass Production Chamber: a testbed for bioregenerative life support studies

The Biomass Production Chamber (BPC) located at Kennedy Space Center, FL, USA provides a large (20 m2 area, 113 m3 vol.), closed environment for crop growth tests for NASA's Controlled Ecological Life Support System (CELSS) program. Since the summer of 1988, the chamber has operated on a near-continuous basis (over 1200 days) without any major failures (excluding temporary power losses). During this time, five crops of wheat (64-86 days each), three crops of soybean (90 to 97 days), five crops of lettuce (28-30 days), and four crops of potato (90 to 105 days were grown, producing 481 kg of dry plant biomass, 196 kg edible biomass, 540 kg of oxygen, 94,700 kg of condensed water, and fixing 739 kg of carbon dioxide. Results indicate that total biomass yields were close to expected values for the given light input, but edible biomass yields and harvest indices were slightly lower than expected. Stand photosynthesis, respiration, transpiration, and nutrient uptake rates were monitored throughout growth and development of the different crops, along with the build-up of ethylene and other volatile organic compounds in the atmosphere. Data were also gathered on system hardware maintenance and repair, as well as person-hours required for chamber operation. Future tests will include long-term crop production studies, tests in which nutrients from waste treatment systems will be used to grow new crops, and multi-species tests.

NASA Center KSC↗

Biomass of the cryptoendolithic microbiota from the Antarctic desert

Extractable lipid phosphate was used to determine the biomass of the cryptoendolithic microbiota that colonizes sandstone rocks in the Ross Desert region of Antarctica. The mean amount of lipid phosphate was 0.053 micromole/cm2 (n = 9), which equals 2.54 g of C per m2 (range, 1.92 to 3.26 g of C per m2) of biomass in the biotic zone of these rocks. The turnover of phospholipids was comparable to that of temperate sediments and soils (t1/2, 6 to 9 days) at 0 degrees C and a light intensity of 305 micromoles of photons per m2 per s, indicating that this was a good method to measure viable biomass. The biomass was 0.3 to 9.6% of the total carbon content of the biotic zone and was about 2 orders of magnitude smaller than the epilithic lichen dry weight at a location some 7 degrees north in latitude. The cryptoendolithic microbiota had a uniform density throughout the biotic zone under the rock surface. The results indicate that the cryptoendolithic microbial biomass is small but viable in this unique, extreme ecosystem.

NASA Discipline Exobiology↗

Biomass accumulation in hydroponically grown sweetpotato in a controlled environment: a preliminary study

In the development of a plant growth model, the assumptions made and the general equations representing an understanding of plant growth are gradually refined as more information is acquired through experimentation. One such experiment that contributed to sweetpotato model development consisted of measuring biomass accumulation of sweetpotato grown in hydroponic culture in a plant growth chamber. Plants were started from fifteen centimeter long 'TU-82-155' sweetpotato vine cuttings spaced 25 cm apart in each of 18 rectangular growing channels (0.15 by 0.15 by 1.2m) in a system designed to use the nutrient film technique (NFT). Each channel contained four plants. The 3.5m by 5.2m plant growth chamber environmental parameters included an 18h photoperiod, 500 micromoles m-2 s-1 of photosynthetic photon flux (PPF), and a diurnal light/dark temperature of 28 degrees C/22 degrees C. The relative humidity was 80 +/- 5% and the CO2 partial pressure was ambient (350 ppm). The nutrient solution contained in 30L reservoirs was a modified half Hoagland's solution with a 1:2.4 N:K ratio and a pH of 6.2. Solution replenishment occurred when the electrical conductivity (EC) level dropped below 1050. Plants were harvested at 15 days after planting (DAP) and weekly thereafter until day 134. By 57 DAP, stems and fibrous roots had acquired 90% of their total dry biomass, while leaves had reached 84% of their maximum dry biomass. Beginning at 64 DAP dry biomass accumulation in the storage roots dominated the increase in dry biomass for the plants. Dry weight of storage roots at 120 DAP was 165 g/plant or 1.1 kg/m2. Resulting growth curves were consistent with the physiological processes occurring in the plant. Results from this study will be incorporated in a plant growth model for use in conjunction with controlled life support systems for long-term manned space missions.

Biomass↗

Evaluation of Stratospheric Intrusions and Biomass Burning Plumes on the Vertical Distribution of Tropospheric Ozone Over the Midwestern U.S.

Naturally occurring ozone rich Stratosphere-to-Troposphere Transport (STT) intrusions and biomass burning plumes reaching the surface can contribute to exceedances of the U.S. National Ambient Air Quality Standards (NAAQS) for ground-level ozone (70 ppbv implemented in 2015). In addition, fires can inject significant pollution into the free troposphere where it can be transported long distances. The combined air quality impacts from these sources on ozone has only been analyzed in a few case studies for the Midwest U.S. Here we study ozone impacts in a major Midwestern city, for the first time in St. Louis, Missouri, using a series of ozonesonde profiles taken during the SEAC4RS (Studies of Emissions and Atmospheric Composition, Clouds and Climate Coupling by Regional Surveys) field campaign in August-September 2013. All ozonesondes showed some enhancements above the background profile levels (~55 ppbv) throughout each tropospheric column. Two models were used to estimate and quantify ozone origins within the columns. A chemical transport model identified enhancements from STT equivalent to 10to15 ppbv over the background with a 10 to15% contribution overall to the column. Two FLEXPART-WRF simulations, one with smoke in the boundary layer and the other with smoke above, identified enhancements from biomass burning equivalent to 10to 80 ppbv. Overall, the total biomass burning contribution is 15to 30% of the total column. Five ozonesondes showed signatures of mixed biomass burning plumes and STT intrusions. During this study period, biomass burning in the western U.S. contributed 70% to ozone enhancements in the total column compared to 3% from the central U.S and 27% from other areas.

National Ambient Air Quality Standards for ground‐↗

An Evaluation of Biomass Burning Aerosol Mass, Extinction, and Size Distribution in Geos Using Observations From Camp2EX

Biomass burning aerosol impacts aspects of the atmosphere and Earth system through radiative forcing, serving as cloud condensation nuclei, and air quality. Despite its importance, the representation of biomass burning aerosol is not always accurate in numerical weather prediction and climate models or reanalysis products. Using observations collected as part of the Cloud, Aerosol and Monsoon Processes Philippines Experiment (CAMP2Ex) in August through October of 2019, aerosol concentration and optical properties are evaluated within the Goddard Earth Observing System (GEOS) and its underlying aerosol module, GOCART. In the operational configuration, GEOS assimilates aerosol optical depth observations at 550 nm to constrain aerosol fields. Particularly for biomass burning aerosol, without the assimilation of aerosol optical depth, aerosol extinction is underestimated compared to observations collected in the Philippines region during the CAMP2Ex campaign. The assimilation process adds excessive amounts of carbon to account for the underestimated extinction, resulting in positive biases in the mass of black and organic carbon, especially within the boundary layer, relative to in situ observations from the Langley Aerosol Research Group Experiment. Counteracting this, GEOS is deficient in sulphate and nitrate aerosol just above the boundary layer. Aside from aerosol mass, extinction within GEOS is a function of ambient relative humidity and an assumed particle size distribution. The relationship between dry and ambient extinction in GEOS reveals that hygroscopic growth is too aggressive within the model for biomass burning aerosol. An additional concern lies in the assumed particle size distribution for GEOS, which has a mode radius that is too small for organic carbon. Variability in the observed particle size distribution for biomass burning aerosol within a single flight also illuminates the fact that a single assumed particle size distribution is not sufficient and that for a proper representation, a more advanced aerosol module with GEOS may be necessary.

Aerosol Mass↗

Biomass Harmonization and SAR Analysis with the Multi-mission Algorithm and Analysis Platform (MAAP)

The Multi‐mission Algorithm and Analysis Platform (MAAP) is a collaborative effort between NASA and the European Space Agency (ESA) to support above ground biomass (AGB) research in an open science framework. MAAP brings together relevant data, algorithms, and computing capabilities in a common cloud environment to address the challenges of sharing and processing data from field, airborne and satellite measurements. MAAP was publicly released in October 2021, providing computing capabilities co-located with the data, a collaborative coding and analysis environment, and a set of interoperable tools and algorithms developed to support the estimation and visualization of data. MAAP has allowed scientists from both North America and Europe to collaborate on the generation and analysis/visualization of data derived from multiple, discipline-adjacent missions in an open, collaborative environment that has reached beyond traditional scientific investigation. MAAP has been used to support multiple scientific activities. To date, existing LiDAR data from multiple platforms has been calibrated with field measurements and combined for more comprehensive and accurate estimates of above ground biomass AGB; these LiDAR platforms include airborne (e.g. LVIS), the International Space Station (NASA’s Global Ecosystem Dynamics Investigation (GEDI), and satellites (e.g. ICESat-2). The current challenge is to effectively and seamlessly combine the aforementioned LiDAR-based data with new data sources such as P-band RADAR from ESA’s upcoming BIOMASS mission, existing ESA Sentinel-1 C-band SAR, and the 30 PB/yr of high cadence global coverage L-band SAR data from the upcoming NASA-ISRO SAR (NISAR) mission. Recent analysis using MAAP merged ICESat-2 and optical data (Harmonized Landsat Sentinel) produced the most comprehensively precise estimate of boreal-wide AGB to date. Another effort using MAAP is the production and open distribution of global comparisons of AGB map estimates, including from ICESat-2 and GEDI, to bolster stakeholder uptake for policy applications. These map estimates will feed into the Intergovernmental Panel on Climate Change (IPCC) database, likely aiding the next Global Carbon Stocktake of the UNFCCC. Furthermore, the biomass retrieval intercomparison exercise BRIX-2 could benefit from the MAAP providing standardized test cases (based on airborne campaign and spaceborne data) allowing the community to develop and apply retrieval algorithms based on these test cases, while forthcoming SAR data training curricula could also use the MAAP as a teaching and learning platform. The MAAP is meeting the challenges inherent in international, open science collaboration and large scale computing with a platform that is entirely open source and cloud native, using open standards for data access, manipulation, protocols, and formats. The MAAP data system consists of a dedicated data store whose data is indexed in an online catalog conforming to established metadata, application programmatic interfaces (APIs), and service interface standards, using an implementation of the open sourced NASA Common Metadata Repository. Federation of user identities allows users from either NASA or ESA to access and consume services from the other using a unified metadata catalog for the data utilized across the ESA and NASA MAAP platforms. Similarly, we are exploring how to increase interoperability to achieve a common approach to packaging, orchestrating and executing algorithms, with interoperable access to data for subsetting, fast browse, and cloud-optimized access, all using interoperable standards such as those from the Open Geospatial Consortium (OGC). Designed for interoperability, ESA and NASA utilize a common architecture for the software platform. It provides a cloud-based algorithm development environment (ADE) that enables scientists to develop algorithms collaboratively with access to the MAAP data catalog as well as other data archives. MAAP provides an Eclipse Che-based ADE supporting both Python and R languages, popular in this biomass community. Algorithms developed and containerized within the ADE can be deployed to run to thousands of computational nodes in the MAAP’s data processing system (DPS), dramatically speeding up processing and giving scientists a rapid, iterative turnaround of results. NASA’s implementation of the DPS is based on the Hybrid Science Data System (HySDS) framework, used by NASA flight projects to produce Earth science standard products.

cloud computing↗

Estimation of old field ecosystem biomass using low altitude imagery

Color-infrared photography was used to evaluate the biomass of experimental plots in an old-field ecosystem that was treated with different levels of waste water from a sewage treatment facility. Cibachrome prints at a scale of approximately 1:1,600 produced from 35 mm color infrared slides were used to analyze density patterns using prepared tonal density scales and multicell grids registered to ground panels shown on the photograph. Correlation analyses between tonal density and vegetation biomass obtained from ground samples and harvests were carried out. Correlations between mean tonal density and harvest biomass data gave consistently high coefficients ranging from 0.530 to 0.896 at the 0.001 significance level. Corresponding multiple regression analysis resulted in higher correlation coefficients. The results of this study indicate that aerial infrared photography can be used to estimate standing crop biomass on waste water irrigated old field ecosystems. Combined with minimal ground truth data, this technique could enable managers of wastewater irrigation projects to precisely time harvest of such systems for maximal removal of nutrients in harvested biomass.

Nor, S. M.↗

Assessing soybean leaf area and leaf biomass by spectral measurements

Red and photographic infrared spectral radiances were correlated with soybean total leaf area index, green leaf area index, chlorotic leaf area index, green leaf biomass, chlorotic leaf biomass, and total biomass. The most significant correlations were found to exist between the IR/red radiance ratio data and green leaf area index and/or green leaf biomass (r squared equals 0.85 and 0.86, respectively). These findings demonstrate that remote sensing data can supply information basic to soybean canopy growth, development, and status by nondestructive determination of the green leaf area or green leaf biomass.

Holben, B. N.↗

Spectral assessment of soybean leaf area and leaf biomass

Red and photographic infrared spectral radiance have been correlated with soybean total leaf area index, green leaf area index, chlorotic area index, green leaf biomass, chlorotic leaf biomass, and total biomass. The most significant correlations were found to exist between the spectral data and green leaf area index and/or green leaf biomass. These findings demonstrate that ground based remote sensing data can supply information basic to soybean canopy growth, development, and status by non-destructive determination of the green leaf area or green leaf biomass.

Holben, B. N.↗

Chemicals from biomass - The U.S. prospects for the turn of the century

Historically, chemicals from biomass have been and are expected to be economical in three major areas: byproducts, specialty items and polymers. Assessments of producing major chemicals from biomass in a processing plant based on the available conversion techniques indicate that they are not economically attractive, with the possible exception of conversion to ammonia and ethanol. The deterrents are the heavy capital investments, dependability of raw material supply and transportation costs for large plants, lack of operation experience, inadaptability of market variations, and competition from petroleum and coal. More importantly, it is also shown that even if chemicals from biomass were economical today, the resultant savings in petroleum would be far less than those achieved through other options available for the utilization of biomass as fuel and structural material. Thus, it is concluded that near-term research and development must be toward improved conversion processes, recovery of valuable products from waste streams at existing plants, more efficient use of biomass of energy and more efficient production of superior material products.

Sarbolouki, M. N.↗

Estimating total standing herbaceous biomass production with LANDSAT MSS digital data

Rangeland biomass data were correlated with spectral vegetation indices, derived from LANDSAT MSS data. LANDSAT data from five range and three other land use sites in Willacv and Cameron Counties were collected on October 17 and December 10, 1975, and on July 31 and September 23, 1976. The overall linear correlation of total standing herbaceous biomass with the LANDSAT derived perpendicular vegetation index was highly significant (r = 0.90**) for these four dates. The standard error of estimate was 722 kg/ha. Biomass data were recorded for two of these range sites for 8 months (March through October) during the 1976 growing season. Standing green biomass accounted for most of the increase in herbage, starting in June and ending about September and October. These results indicate that satellite data may be useful for the estimation of total standing herbaceous biomass production that could aid range managers in assessing range condition and animal carrying capacities of large and inaccessible range holdings.

Richardson, A. J.↗

Development of visible/infrared/microwave agriculture classification and biomass estimation algorithms, volume 2

Agricultural crop classification models using two or more spectral regions (visible through microwave) were developed and tested and biomass was estimated by including microwave with visible and infrared data. The study was conducted at Guymon, Oklahoma and Dalhart, Texas utilizing aircraft multispectral data and ground truth soil moisture and biomass information. Results indicate that inclusion of C, L, and P band active microwave data from look angles greater than 35 deg from nadir with visible and infrared data improved crop discrimination and biomass estimates compared to results using only visible and infrared data. The active microwave frequencies were sensitive to different biomass levels. In addition, two indices, one using only active microwave data and the other using data from the middle and near infrared bands, were well correlated to total biomass.

Rosenthal, W. D.↗

Biomass recycle as a means to improve the energy efficiency of CELSS algal culture systems

Algal cultures can be very rapid and efficient means to generate biomass and regenerate the atmosphere for closed environmental life support systems. However, as in the case of most higher plants, a significant fraction of the biomass produced by most algae cannot be directly converted to a useful food product by standard food technology procedures. This waste biomass will serve as an energy drain on the overall system unless it can be efficiently recycled without a significant loss of its energy content. Experiments are reported in which cultures of the alga Scenedesmus obliquus were grown in the light and at the expense of an added carbon source, which either replaced or supplemented the actinic light. As part of these experiments, hydrolyzed waste biomass from these same algae were tested to determine whether the algae themselves could be made part of the biological recycling process. Results indicate that hydrolyzed algal (and plant) biomass can serve as carbon and energy sources for the growth of these algae, suggesting that the efficiency of the closed system could be significantly improved using this recycling process.

Radmer, R.↗

Remote sensing of biomass of salt marsh vegetation in France

Spectral data (gathered using a hand-held radiometer) and harvest data were collected from four salt marsh vegetation types in Brittany, France, to develop equations predicting live aerial biomass from spectral measurements. Remote sensing estimates of biomass of the general salt marsh community (GSM) and of Spartina alterniflora can be obtained throughout the growing season if separate biomass prediction equations are formulated for different species mixtures (for the GSM) and for different canopy types (for S. alterniflora). Results suggest that remote sensing will not be useful for predicting Halimione portulacoides biomass, but can be used to estimate Puccinellia maritima biomass early in the growing season.

Gross, M. F.↗

Biomass production and nitrogen dynamics in an integrated aquaculture/agriculture system

A combined aquaculture/agriculture system that brings together the three major components of a Controlled Ecological Life Support System (CELSS) - biomass production, biomass processing, and waste recycling - was developed to evaluate ecological processes and hardware requirements necessary to assess the feasibility of and define design criteria for integration into the Kennedy Space Center (KSC) Breadboard Project. The system consists of a 1 square meter plant growth area, a 500 liter fish culture tank, and computerized monitoring and control hardware. Nutrients in the hydrophonic solution were derived from fish metabolites and fish food leachate. In five months of continuous operation, 27.0 kg of lettuce tops, 39.9 kg of roots and biofilm, and 6.6 kg of fish (wet weights) were produced with 12.7 kg of fish food input. Based on dry weights, a biomass conversion index of 0.52 was achieved. A nitrogen budget was derived to determine partitioning of nitrogen within various compartments of the system. Accumulating nitrogen in the hypoponic solution indicated a need to enlarge the plant growth area, potentially increasing the biomass production and improving the biomass conversion index.

Owens, L. P.↗

Estimating slash pine biomass using radar backscatter

L-band HV multiple-incidence-angle aircraft synthetic aperture radar (SAR) data were analyzed in relation to average stand biomass, basal area, and tree height for 55 slash pine plantations located in northern Florida. This information was used to develop a system of equations to predict average stand biomass as a function of L-band (24.5-cm) radar backscatter. The system of equations developed in this study using three-stage least-squares and combinatorial screening accounted for 97 percent of the variability observed in average stand biomass per hectare. When applied to an independent data set, the biomass equations had an average bias of less than 1 percent with a standard error of approximately 3 percent. These results indicate that future Shuttle Imaging Radar Systems (e.g., SIR-C, which will have cross-polarized radar sensors) should be able to obtain better estimates of forest biomass than were obtained with previous satellite radar missions, which utilized only HH-polarized SAR data.

Hussin, Yousif Ali↗