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At least 19 records

Microbial biomass and productivity in seagrass beds

Different methods for measuring the rates of processes mediated by bacteria in sediments and the rates of bacterial cell production have been compared. In addition, net production of the seagrass Zostera capricorni and bacterial production have been compared and some interrelationships with the nitrogen cycle discussed. Seagrass productivity was estimated by measuring the plastochrone interval using a leaf stapling technique. The average productivity over four seasons was 1.28 +/- 0.28 g C m-2 day-1 (mean +/- standard deviation, n = 4). Bacterial productivity was measured five times throughout a year using the rate of tritiated thymidine incorporated into DNA. Average values were 33 +/- 12 mg C m-2 day-1 for sediment and 23 +/- 4 for water column (n = 5). Spatial variability between samples was greater than seasonal variation for both seagrass productivity and bacterial productivity. On one occasion, bacterial productivity was measured using the rate of 32P incorporated into phospholipid. The values were comparable to those obtained with tritiated thymidine. The rate of sulfate reduction was 10 mmol SO4(-2) m-2 day-1. The rate of methanogenesis was low, being 5.6 mg CH4 produced m-2 day-1. A comparison of C flux measured using rates of sulfate reduction and DNA synthesis indicated that anaerobic processes were predominant in these sediments. An analysis of microbial biomass and community structure, using techniques of phospholipid analysis, showed that bacteria were predominant members of the microbial biomass and that of these, strictly anaerobic bacteria were the main components. Ammonia concentration in interstitial water varied from 23 to 71 micromoles. Estimates of the amount of ammonia required by seagrass showed that the ammonia would turn over about once per day. Rapid recycling of nitrogen by bacteria and bacterial grazers is probably important.

NASA Program CELSS↗

Electron Microscopy of Microbial Biomass

As humans move to long term stays in space their microbes move with them. This poses problems as the microbes can colonize various life support systems, creating biofilms that can grow and spread, which in turn causes obstructions and ultimately degrades function of these systems. MSFC currently has no protocol in place for the Scanning Electron Microscope (SEM) imaging of microbes.

Samantha Marcella↗

Transitory Microbial Habitat in the Hyperarid Atacama Desert

Traces of life are nearly ubiquitous on Earth. However, a central unresolved question is whether these traces always indicate an active microbial community or whether, in extreme environments, such as hyperarid deserts, they instead reflect just dormant or dead cells. Although microbial biomass and diversity decrease with increasing aridity in the Atacama Desert, we provide multiple lines of evidence for the presence of an at times metabolically active, microbial community in one of the driest places on Earth. We base this observation on four major lines of evidence: (i) a physico-chemical characterization of the soil habitability after an exceptional rain event, (ii) identified biomolecules indicative of potentially active cells [e.g., presence of ATP, phospholipid fatty acids (PLFAs), metabolites, and enzymatic activity], (iii) measurements of in situ replication rates of genomes of uncultivated bacteria reconstructed from selected samples, and (iv) microbial community patterns specific to soil parameters and depths. We infer that the microbial populations have undergone selection and adaptation in response to their specific soil microenvironment and in particular to the degree of aridity. Collectively, our results highlight that even the hyperarid Atacama Desert can provide a habitable environment for microorganisms that allows them to become metabolically active following an episodic increase in moisture and that once it decreases, so does the activity of the microbiota. These results have implications for the prospect of life on other planets such as Mars, which has transitioned from an earlier wetter environment to today’s extreme hyperaridity.

Schulze-Makuch, Dirk↗

Microbial Habitability in Gale Crater: Sample Analysis at Mars (SAM) Instrument Detection of Microbial Essential Carbon and Nitrogen

Chemical analyses of Mars soils and sediments from previous landed missions have demonstrated that Mars surface materials possessed major (e.g., P, K, Ca, Mg, S) and minor (e.g., Fe, Mn, Zn, Ni, Cl) elements essential to support microbial life. However, the detection of microbial essential organic-carbon (C) and nitrate have been more elusive until the Mars Science Laboratory (MSL) rover mission. Nitrate and organic-C in Gale Crater, Mars have been detected by the Sample Analysis at Mars (SAM) instrument onboard the MSL Curiosity rover. Eolian fines and drilled sedimentary rock samples were heated in the SAM oven from approximately 30 to 860 degrees Centigrade where evolved gases (e.g., nitrous oxide (NO) and CO2) were released and analyzed by SAM’s quadrupole mass spectrometer (MS). The temperatures of evolved NO was assigned to nitrate while evolved CO2 was assigned to organic-C and carbonate. The CO2 releases in several samples occurred below 450 degrees Centigrade suggesting organic-C dominated in those samples. As much as 7 micromoles NO3-N per gram and 200 micromoles CO2-C per gram have been detected in the Gale Crater materials. These N and C levels coupled with assumed microbial biomass (9 x 10 (sup -7) micrograms per cell) C (0.5 micrograms C per micrograms cell) and N (0.14 micrograms N per micrograms cell) requirements, suggests that less than 1 percent and less than 10 percent of Gale Crater C and N, respectively, would be required if available, to accommodate biomass requirements of 1 by 10 (sup 5) cells per gram sediment. While nitrogen is the limiting nutrient, the potential exists that sufficient N and organic-C were present to support limited heterotrophic microbial populations that may have existed on ancient Mars.

Sutter, B.↗

Distinguishing Indigenous from Contaminating Microorganisms in Rock Samples from a Deep Au Mine in South Africa

The concentration and distribution of microbial biomass within deep subsurface rock strata is not well known To date, most analyses are from water samples and a few cores. Hand samples, block samples and cores from an actively mined Carbon Leader ore zone at 3.2 kilometers depth were collected for microbial analyses. The Carbon Leader was comprised of quartz, S-bearing aromatic hydrocarbons, Fe(III) oxyhydroxides, sulfides, uraninite, Au and minor amounts of sulfate. The porosity of the ore was 1% and the maximum pore throat diameter was less than 0.1 microns; whereas, the porosity of the adjacent quartzite was .02 to .9% with a maximum pore throat diameter of 0.9 microns. Rhodamine dye, fluorescent microspheres, microbial enrichments, autoradiography, phospholipid fatty acid (PLEA) and 16S rDNA analyses were performed on these rock samples and the mining water. The date indicate that the levels of solute contamination less than 0.01% for pared rock samples. Despite this low level of contamination, PLEA, microbial enrichment, DNA and tracer analyses and calculations indicate that most of the viable microorganisms in the Carbon Leader represent gram negative aerobic heterotrophs and ammonia oxidizers that are phylogenetically identical or closely related to service water microorganisms. These microbial contaminants probably infiltrated the low permeability rock through mining-induced microfractures. Geochemical data also detected drilling water in a fault zone approx. 1 meter behind the rock face encountered during coring. The mining induced macrofractures that are common at these great depths act as pathways for the drilling water borne microorganisms into the lower temperature zone that extends several meters into rock strata from the rock face. Combined PLEA and T- RFLP analyses of the service water and Carbon Leader samples indicate that the concentration of indigenous microorganisms was less than 10(exp 2) cells/gram. Such a low concentrations result from the submicron pore throat diameters. PLFA. SO4-35 autoradiography and tracer analyses indicate that the bounding quartzite contains thermophilic sulfate reducing bacteria at 10(exp 3) cells/gram that are not attributable to drilling water contamination. The microorganisms may be surviving on sulfate generated by oxidation of sulfide by radiolytic reactions resulting from the high U concentration in the ore zone. The presence of up to 8,000 ppm of Fe(III) oxyhydroxides in the host rock will also act to recycle sulfide generated by the sulfate reducing bacteria into sulfate. The activity of these sulfate-reducing bacteria may be enhanced by mining induced fracturing which can propagate up to 40 meters into virgin rock where the temperatures are ca. 50 C, and decrepitate of sulfate rich fluid inclusions. In ultra deep mines, judicious application of tracers and multiple microbial characterization techniques can distinguish microbial contamination caused by the near field fracturing and drilling water migration from the indigenous microbial communities in rock strata. The importance of far field fracturing on indigenous microbial communities, however, remains unknown.

Onstott, T. C.↗

Nitrogen transformations following tropical forest felling and burning on a volcanic soil

Nitrogen transformations and loss were measured following forest clearing in a relatively fertile tropical forest site. Nitrogen mineralization, nitrification, and amounts of ammonium and nitrate increased substantially in surface soils during the 6 mo following burning, then returned to background levels. The nitrogen content of microbial biomass declined to half its original value 6 mo after clearing and remained low in the cleared sites. Plant uptake of nitrogen was substantial on cleared plots (50 g/sq m), but it accounted for only 18 percent of N-15 label added to field plots. MIcrobial immobilization of N-15 was small relative to that in a cleared temperate site, and measurements of denitrification potentials suggested that relatively little mineralized nitrogen was lost to the atmosphere. Substantial amounts of nitrogen (40-70 g/sq m) were retained as exchangeably bound nitrate deep in the soils of a cleared plot on which revegetation was prevented; this process accounted for 12 percent of the N-15 label added to field plots.

Matson, Pamela A.↗

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↗

Applications of mutant yeast strains with low glycogen storage capability

Several strains of Hansenula polymorpha were selected for possible low glycogen storage characteristics based on a selective I2 staining procedure. The levels of storage carbohydrates in the mutant strains were found to be 44-70% of the levels in the parent strain for cultures harvested in stationary phase. Similar differences generally were not found for cells harvested in exponential phase. Yeast strains deficient in glycogen storage capability are valuable in increasing the relative protein value of microbial biomass and also may provide significant cost savings in substrate utilization in fermentative processes.

Petersen, G. R.↗

Soil nitrogen cycling and nitrous oxide flux in a Rocky Mountain Douglas-fir forest - Effects of fertilization, irrigation and carbon addition

Nitrous oxide fluxes and soil nitrogen transformations were measured in experimentally-treated high elevation Douglas-fir forests in northwestern New Mexico. On an annual basis, forests that were fertilized with 200 kg N/ha emitted an average of 0.66 kg/ha of N2O-N, with highest fluxes occurring in July and August when soils were both warm and wet. Control, irrigated, and woodchip treated plots were not different from each other, and annual average fluxes ranged from 0.03 to 0.23 kg/ha. Fertilized soil mineralized 277 kg/ha per year in contrast to 18 kg/ha per year in control plots. Relative recovery of (N-15)H4-N applied to soil in laboratory incubations was principally in the form of NO3-N in the fertilized soils, while recovery was mostly in microbial biomass-N in the other treatments. Fertilization apparently added nitrogen that exceeded the heterotrophic microbial demand, resulting in higher rates of nitrate production and higher nitrous oxide fluxes. Global inputs of nitrogen into forests are not currently contributing significantly to the increasing concentrations of nitrous oxide in the atmosphere.

Matson, Pamela A.↗

Soil nitrogen cycling and nitrous oxide flux in a Rocky Mountain Douglas-fir forest - Effects of fertilization, irrigation and carbon addition

Nitrous oxide fluxes and soil nitrogen transformations were measured in experimentally-treated high elevation Douglas-fir forests in northwestern New Mexico, USA. On an annual basis, forests that were fertilized with 200 kg N/ha emitted an average of 0.66 kg/ha of N2O-N, with highest fluxes occurring in July and August when soils were both warm and wet. Control, irrigated, and woodchip treated plots did not differ, and annual average fluxes ranged from 0.03 to 0.23 kg/ha. Annual net nitrogen mineralization and nitrate production were estimated in soil and forest floor using in situ incubations; fertilized soil mineralized 277 kg/ha/y in contrast to 18 kg/ha/y in control plots. Relative recovery of 15NH4-N applied to soil in laboratory incubations was principally in the form of NO3-N in the fertilized soils, while recovery was mostly in microbial biomass-N in the other treatments. Fertilization apparently added nitrogen that exceeded the heterotrophic microbial demand, resulting in higher rates of nitrate production and higher nitrous oxide fluxes. Despite the elevated nitrous oxide emission resulting from fertilization, we estimate that global inputs of nitrogen into forests are not currently contributing significantly to the increasing concentrations of nitrous oxide in the atmosphere.

Matson, Pamela A.↗

Biomass Burning and the Production of Greenhouse Gases

Biomass burning is a source of greenhouse gases, carbon dioxide, methane, and nitrous oxide. In addition, biomass burning is a source of chemically active gases, including carbon monoxide, nonmethane hydrocarbons, and nitric oxide. These gases, along with methane, lead to the chemical production of tropospheric ozone (another greenhouse gas) as well as control the concentration of the hydroxyl radical, which regulates the lifetime of almost every atmospheric gas. Following biomass burning, biogenic emissions of nitrous oxide, nitric oxide, and methane are significantly enhanced. It is hypothesized that enhanced postburn biogenic emissions of these gases are related to fire-induced changes in soil chemistry and/or microbial ecology. Biomass burning, once believed to be a tropical phenomenon, has been demonstrated by satellite imagery to also be a regular feature of the world's boreal forests. One example of biomass burning is the extensive 1987 fire that destroyed more than 12 million acres of boreal forest in the People's Republic of China and across its border in the Soviet Union. Recent estimates indicate that almost all biomass burning is human-initiated and that it is increasing with time. With the formation of greenhouse and chemically active gases as direct combustion products and a longer-term enhancement of biogenic emissions of gases, biomass burning may be a significant driver for global change.

Levine, Joel S.↗

Recycling crop residues for use in recirculating hydroponic crop production

As part of bioregenerative life support feasibility testing by NASA, crop residues are being used to resupply elemental nutrients to recirculating hydroponic crop production systems. Methods for recovering nutrients from crop residues have evolved from water soaking (leaching) to rapid aerobic bioreactor processing. Leaching residues recovered the majority of elements but it also recovered significant amounts of soluble organics. The high organic content of leachates was detrimental to plant growth. Aerobic bioreactor processing reduced the organic content ten-fold, which reduced or eliminated phytotoxic effects. Wheat and potato production studies were successful using effluents from reactors having with 8- to 1-day retention times. Aerobic bioreactor effluents supplied at least half of the crops elemental mass needs in these studies. Descriptions of leachate and effluent mineral content, biomass productivity, microbial activity, and nutrient budgets for potato and wheat are presented.

NASA Center KSC↗

Pick-and-Eat Salad-Crop Productivity, Nutritional Value, and Acceptability to Supplement the ISS Food System

The capability to grow nutritious, palatable food for crew consumption during spaceflight has the potential to provide health-promoting, bioavailable nutrients, enhance the dietary experience, and reduce launch mass as we move toward longer-duration missions. However, studies of edible produce during spaceflight have been limited, leaving a significant knowledge gap in the methods required to grow safe, acceptable, nutritious crops for consumption in space. Researchers from Kennedy Space Center, Johnson Space Center, Purdue University and ORBITEC have teamed up to explore the potential for plant growth and food production on the International Space Station (ISS) and future exploration missions. KSC, Purdue, and ORBITEC bring a history of plant and plant-microbial interaction research for ISS and for future bioregenerative life support systems. JSC brings expertise in Advanced Food Technology (AFT), Behavioral Health and Performance (BHP), and statistics. The Veggie vegetable-production system on the ISS offers an opportunity to develop a pick-and-eat fresh vegetable component to the ISS food system as a first step to bioregenerative supplemental food production. We propose growing salad plants in the Veggie unit during spaceflight, focusing on the impact of light quality and fertilizer formulation on crop morphology, edible biomass yield, microbial food safety, organoleptic acceptability, nutritional value, and behavioral health benefits of the fresh produce. The first phase of the project will involve flight tests using leafy greens, with a small Chinese cabbage variety, Tokyo bekana, previously down selected through a series of research tests as a suitable candidate. The second phase will focus on dwarf tomato. Down selection of candidate varieties have been performed, and the dwarf cultivar Red Robin has been selected as the test crop. Four light treatments and three fertilizer treatments will be tested for each crop on the ground, to down select to two light treatments and one fertilizer treatment to test on ISS. Our work will help define light colors, levels, and horticultural best practices to achieve high yields of safe, nutritious leafy greens and tomatoes to supplement a space diet of prepackaged food. Our final deliverable will be the development of growth protocols for these crops in a spaceflight vegetable production system. With this work, and potentially with other pending joint projects, we will continue the synergistic research to help close gaps in the human research roadmap, and enable humans to venture to Mars and beyond. This research was co-funded by the Human Research Program and Space Biology (MTL1075) in the ILSRA 2015 NRA call.

Behavioral Health↗

Forearc Carbon Sink Reduces Long-Term Volatile Recycling Into the Mantle

Carbon and other volatiles are transported from Earth’s surface into the mantle at subduction margins. The efficiency of this transfer has profound implications for the nature and scale of geochemical heterogeneities in Earth’s deep (mantle) and shallow (crustal) reservoirs, as well as Earth’s oxidation state. However, the proportion of volatiles released in the forearc and backarc are not well constrained compared to fluxes from the volcanic front. Here, we use helium and carbon isotope data from deeply sourced springs along two cross-arc transects to show that ~91% of carbon released from the slab/mantle beneath the Costa Rica forearc is sequestered within the crust by calcite deposition, and an additional ~3% is incorporated into biomass through microbial chemolithoautotrophy. We estimate that ~1.2 × 10(exp 8) to 1.3 × 10(exp 10) mol CO2/yr are released from the slab beneath the forearc, resulting in up to ~19% less carbon being transferred to Earth’s deep mantle than previously estimated.

Carbon cycle↗

Rapid System to Quantitatively Characterize the Airborne Microbial Community

Bioaerosols have been linked to a wide range of different allergies and respiratory illnesses. Currently, microorganism culture is the most commonly used method for exposure assessment. Such culture techniques, however, generally fail to detect between 90-99% of the actual viable biomass. Consequently, an unbiased technique for detecting airborne microorganisms is essential. In this Phase II proposal, a portable air sampling device his been developed for the collection of airborne microbial biomass from indoor (and outdoor) environments. Methods were evaluated for extracting and identifying lipids that provide information on indoor air microbial biomass, and automation of these procedures was investigated. Also, techniques to automate the extraction of DNA were explored.

Macnaughton, Sarah J.↗

Deep-Sea Hydrothermal-Vent Sampler

An apparatus is being developed for sampling water for signs of microbial life in an ocean hydrothermal vent at a depth of as much as 6.5 km. Heretofore, evidence of microbial life in deep-sea hydrothermal vents has been elusive and difficult to validate. Because of the extreme conditions in these environments (high pressures and temperatures often in excess of 300 C), deep-sea hydrothermal- vent samplers must be robust. Because of the presumed low density of biomass of these environments, samplers must be capable of collecting water samples of significant volume. It is also essential to prevent contamination of samples by microbes entrained from surrounding waters. Prior to the development of the present apparatus, no sampling device was capable of satisfying these requirements. The apparatus (see figure) includes an intake equipped with a temperature probe, plus several other temperature probes located away from the intake. The readings from the temperature probes are utilized in conjunction with readings from flowmeters to determine the position of the intake relative to the hydrothermal plume and, thereby, to position the intake to sample directly from the plume. Because it is necessary to collect large samples of water in order to obtain sufficient microbial biomass but it is not practical to retain all the water from the samples, four filter arrays are used to concentrate the microbial biomass (which is assumed to consist of particles larger than 0.2 m) into smaller volumes. The apparatus can collect multiple samples per dive and is designed to process a total volume of 10 L of vent fluid, of which most passes through the filters, leaving a total possibly-microbe-containing sample volume of 200 mL remaining in filters. A rigid titanium nose at the intake is used for cooling the sample water before it enters a flexible inlet hose connected to a pump. As the water passes through the titanium nose, it must be cooled to a temperature that is above a mineral-precipitation temperature of 100 C but below the upper working temperature (230 C) of switching valves and tubes in the apparatus. The sample water then passes into a manifold tube, from whence the switching valves can direct the water through either a bypass tube or any one of the filter arrays, without contamination from a previous sample. Each filter array consists of series of filters having pore sizes decreasing in the direction of flow: 90-, 60-, 15-, and 7-micron prefilters and a large-surface-area 0.2-micron collection filter. All the filter taps are located between the intake and the bypass tube so that each time the bypass tube is used, the entire manifold tube is flushed as well.

Behar, Alberto E.↗

Microgreens Root Zone/Shoot Zone Partitioned Planting Box

To enable sustainable food production in future human exploration missions, plant growth is being studied by the Space Crop Production Team at KSC. Microgreens are good candidates for food supplements and contain specific nutrients that are lacking in the prepackaged diet, including vitamin C and vitamin K. Because they are densely sown, typical growth methods do not allow the ability to distinguish between the levels of evaporation from the rooting substrate and transpiration from the leaves. With larger plants, the root and shoot zone can be separated to distinguish these fluxes and accurate transpiration measurements of plant canopies are feasible. Furthermore, separation of the root and shoot zone may also be beneficial when harvesting microgreens in microgravity as it may reduce microbial contamination of the edible biomass by the roots, which have high microbial loads. Using a root and shoot separator box when harvesting may help with microbial contamination, but these tiny plants are challenging to handle in microgravity, so harvest management remains an open question. The innovation proposed here is a microgreen root/shoot partitioned planting box, which offers a solution to these challenges with accurate gas exchange measurements and a safe microgreen harvest in low gravity environments. Being able to measure transpiration of a microgreen canopy will be important for modeling plant growth in reduced gravity environments, so the first objective was to develop a planting unit with a seal between the root zone and the shoot zone. With an unsuitable harvesting technique, freshly harvested microgreens may add debris to the cabin, so the second objective was to test different harvesting techniques and management approaches associated with this innovation. These two objectives were pursued in parallel since many goals were the same: develop a planting unit that 1) separates the shoots from the roots, 2) allows acceptable germination rate, and 3) allows for seedlings to emerge and develop. What differed was the need to have a seal, which was only applicable for our gas exchange goal, and the need to have an embedded harvesting mechanism and bagging method which only applied to our harvesting goal. Testing of the various harvesting mechanisms and bagging methods was performed during a series of parabolic flights. All parabolic flight procedures took place inside a rented secondary containment chamber (e.g., glovebox) that was developed by the University of Louisville specifically for experiments involving fluids and other materials that may become airborne during reduced gravity flight. Three different harvesting methods and two different bagging collection methods were tested for microgreens. A third bagging method was initially tested but found to be unsuitable. Human factors were also taken into consideration, to identify which harvest and bagging collection methods would be easiest to use with favorable results in microgravity. Three parabolic flight tests were performed in total, one in November 2021 and two in December 2021.

Gioia Massa↗

Psychrophilic Biomass Producers in the Trophic Chain of the Microbial Community of Lake Untersee, Antarctica

The study of photosynthetic microorganisms from the Lake Untersee samples showed dispersed distribution of phototrophs within ~80 m water column. Lake Untersee represents a unique ecosystem that experienced complete isolation: sealed by the Anuchin Glacier for many millennia. Consequently, its biocenosis has evolved over a significant period of time without exchange or external interaction with species from other environments. The major producers of organic matter in Lake Untersee are represented by phototrophic and chemolithotrophic microorganisms. This is the traditional trophic scheme for lacustrine ecosystems on Earth. Among the phototrophs, diatoms were not found, which differentiates this lake from other known ecosystems. The dominant species among phototrophs was Chlamydomonas sp. with typical morphostructure: green chloroplasts, bright red round spot, and two polar flagella near the opening. As expected, the physiology of studied phototrophs was limited by low temperature, which defined them as obligate psychrophilic microorganisms. By the quantity estimation of methanogenesis in this lake, the litho-autotrophic production of organic matter is competitive with phototrophic production. However, pure cultures of methanogens have not yet been obtained. We discuss the primary producers of organic matter and the participation of our novel psychrophilic homoacetogen into the litho-autotrophic link of biomass production in Lake Untersee.

Pikuta, Elena V.↗