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At least 1,621 records · Page 90

The microbiologist's guide to metaproteomics

Metaproteomics is an emerging approach for studying microbiomes, offering the ability to characterize proteins that underpin microbial functionality within diverse ecosystems. As the primary catalytic and structural components of microbiomes, proteins provide unique insights into the active processes and ecological roles of microbial communities. By integrating metaproteomics with other omics disciplines, researchers can gain a comprehensive understanding of microbial ecology, interactions, and functional dynamics. This review, developed by the Metaproteomics Initiative (www.metaproteomics.org), serves as a practical guide for both microbiome and proteomics researchers, presenting key principles, state-of-the-art methodologies, and analytical workflows essential to metaproteomics. Topics covered include experimental design, sample preparation, mass spectrometry techniques, data analysis strategies, and statistical approaches.

bioinformatics↗

Elucidating algal-bacterial community interactions by tracking volatile biomarkers (Final Report)

This feasibility study project aimed to lay further groundwork for studying and understanding how microorganisms interact with each other at the molecular level using model algal-bacterial co-cultures. A better understanding of such interactions is critical to optimizing and operating biotic platforms for a number of applications, including bioenergy, algal bioproducts and agriculture, and carbon capture. Here we explored the feasibility of characterizing and following these interactions by identifying and tracking volatile metabolites and biomarkers in the complex gaseous headspace of such cultures. We set up an experimental platform using a co-culture of Phaeodactylum tricornutum and Marinobacter subspecies 3-2 as a model system, used solid-phase microextraction fibers for volatile collection at various time points throughout culture growth, and employed gas chromatography mass spectrometry-based instrumentation available at LLNL for compound identification and quantification. We found that it is feasible to determine volatile profiles from the growth of Phaeodactylum tricornutum and Marinobacter subspecies 3-2 using our methodology and monitor profile changes over time. We also observed that volatiles distinguishing between cultures of the individual species and the co-cultures could be detected. We expect that results of this project will feed into the ongoing LLNL’s DOE BER funded Biofuels Scientific Focus Area (SFA) on algal-bacterial interactions and pave the way for future expanded volatile studies supported by DOE. The approach and technologies developed here would also be applicable and transferable to other microbial communities under study, such as cyanobacteria, rhizosphere communities and biofilms.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Point-scale organic-matter decomposition in streambeds is weakly associated with reach-scale respiration

Stream and river ecosystems play a central role in the movement and decomposition of particulate organic matter, serving as a conduit between terrestrial hillslopes and coastal environments. Microbial-catalyzed decomposition generates simpler organic molecules that fuel respiration, often in the sediments of these ecosystems. However, the degree of connection between sediment-associated respiration (ER??d) and organic-matter decomposition remains poorly understood. How that relationship compares to decomposition’s relationship with whole ecosystem (ERtot) and water column (ERwc) respiration is also not clear. We examined the link between particulate organic matter decomposition—using cellulose-based cotton strips as a standardized substrate—and all three components of respiration across 48 sites in the environmentally diverse Yakima River Basin (Washington State, USA). We hypothesized that decomposition within sediments would be most strongly related to ERsed, but decomposition rates were more closely associated with ERtot, with little connection to ERsed or ERwc. This suggests that particulate organic matter decomposition within stream/river sediments reflects integrated system respiration rather than processes confined to sediments or the water column alone. Further, across the basin, decomposition rates nearly spanned the previously reported global range for streams and rivers and were best explained by total dissolved nitrogen (TDN), sediment grain size, and aridity of the upstream drainage area. These results highlight the strong influence of land cover and basin-scale biophysical variation on sediment-associated decomposition processes and indicate that mechanistic models of organic matter decomposition in streams/rivers should account for coupled sediment–water–land interactions.

Stegen, James C. (ORCID:0000000191357424)↗

International Space Station USOS Potable Water Dispenser On-Orbit Functionality Versus Design

The International Space Station (ISS) currently provides potable water dispensing for rehydrating crewmember food and drinking packages. There is one system located in the United States On-orbit Segment (USOS) and one system in the Russian Segment. Shuttle mission STS-126 delivered the USOS Potable Water Dispenser (PWD) to ISS on ULF2; subsequent activation occurred on November 2008. The PWD is capable of supporting an ISS crew of six, but nominally supplies only half this crew size. The PWD design provides incremental quantities of hot and ambient temperature potable water to US food and beverage packages. PWD receives iodinated water from the US Water Recovery System (WRS) Fuel Cell Water Bus, which feeds from the Water Processing Assembly (WPA). The PWD removes the biocidal iodine to make the water potable prior to dispensing. A heater assembly contained within the unit supplies up to 2.0 L of hot water (65 to 93 ?C) every 30 min. During a single meal, this quantity of water supports three to four crewmembers? food rehydration and beverages. The unit design has a functional life expectancy of 10 years, with replacement of limited life items, such as filters. To date, the PWD on-orbit performance is acceptable. Since activation of the PWD, there were several differences between on-orbit functionality and expected performance of hardware design. The comparison of on-orbit functionality to performance of hardware design is discussed for the following key areas: 1) microbial contamination, 2) no-dispense and water leakage scenarios, and 3) under-dispense scenarios.

Toon, Katherine P.↗

Impact of UV-light and pH on the Fate of Tc, I, and U in Wetlands at Savannah River Site - 20230

The Savannah River Site (SRS) is one of the major nuclear facilities owned by the U.S Department of Energy. During the Cold War, these facilities produced large amounts of radioactive and hazardous waste. On site, three unlined seepage basins in the F-Area received approximately 1.8 billion gallons of low-level waste containing nitric acid, radionuclides, and dissolved metals due to plutonium and tritium production operations. The acidic nature of the waste created a source of relatively mobile radionuclides below the basins. Radionuclides previously disposed of within the F-Area, including uranium isotopes (U), technetium-99 (Tc-99), and iodine-129 (I-129), are moving with groundwater towards Four Mile Branch Wetland, where they are subsequently upwelling and interacting with natural organic matter (NOM). Many environmental factors including redox conditions, porewater composition, pH, light, and temperature affect the degradation of organic matter as well as interactions with Tc-99, U, and I-129. In particular, the high concentrations of nitrate from the acidic waste may increase the formation of reactive oxygen species (ROS) that impact both degradation of NOM and behavior of contaminants. In the presence of sunlight, I-129, Tc-99, and U speciation and complexation behavior may be affected by ROS and NOM degradation products in addition to the factors generally considered in subsurface systems in the absence of light. This research aims to determine whether the interactions between radionuclides, NOM, and nitrate affect the fate of I-129, Tc-99, and U and which processes are controlling their behavior. Experiments were conducted at variable pH in the presence of NaNO{sub 3} in order to determine the impact of light and pH on NOM degradation and to evaluate the impact on the fate of contaminants of concern. Soil samples high in NOM collected from two sites in the Southeastern United States (Savannah River Site and the Everglades) were studied. Batch experiments were conducted with NOM and the following aqueous contaminants, U, I-129, Tc-99 with exposure to ultraviolet (UV) light in an environmental chamber. This study was conducted in a sterile environment in order to exclude the potential for microbial degradation of organic matter. Results indicated photodegradation of NOM and significant interaction of radionuclides with NOM. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Near‐Surface Hydrology and Soil Properties Drive Heterogeneity in Permafrost Distribution, Vegetation Dynamics, and Carbon Cycling in a Sub‐Arctic Watershed

Abstract Discontinuous permafrost environments exhibit strong spatial heterogeneity at scales too small to be driven by weather forcing or captured by Earth System Models. Here we analyze effects of observed spatial heterogeneity in soil and vegetation properties, hydrology, and thermal dynamics on ecosystem carbon dynamics in a watershed on the Seward Peninsula in Alaska. We apply a Morris global sensitivity analysis to a process‐rich, successfully tested terrestrial ecosystem model (TEM), ecosys , varying soil properties, boundary conditions, and weather forcing. We show that landscape heterogeneity strongly impacts soil temperatures and vegetation composition. Snow depth, O‐horizon thickness, and near‐surface water content, which vary at scales of O(m), control the soil thermal regime more than an air temperature gradient corresponding to a 140 km north–south distance. High shrub productivity is simulated only in talik (perennially unfrozen) soils with high nitrogen availability. Through these effects on plant and permafrost dynamics, landscape heterogeneity impacts ecosystem productivity. Simulations with near‐surface taliks have higher microbial respiration (by 78.0 gC m −2 yr −1 ) and higher net primary productivity (by 104.9 gC m −2 yr −1 ) compared to runs with near‐surface permafrost, and simulations with high shrub productivity have outlying values of net carbon uptake. We explored the prediction uncertainty associated with ignoring observed landscape heterogeneity, and found that watershed net carbon uptake is 60% larger when heterogeneity is accounted for. Our results highlight the complexity inherent in discontinuous permafrost environments and demonstrate that missing representation of subgrid heterogeneity in TEMs could bias predictions of high‐latitude carbon budget.

54 ENVIRONMENTAL SCIENCES↗

Improved chemical and isotopic labeling of biomembranes in Bacillus subtilis by leveraging CRISPRi inhibition of beta-ketoacyl-ACP synthase (fabF)

Assessing the structure of living microbial cell membranes is a challenging analytical goal. The cell membrane is defined by its transverse structure, an approximately 5 nm-thick selectively permeable bilayer that serves many important cellular functions. Compositionally complex, dynamic, and organized in both the transverse and lateral dimensions, understanding the cell membrane structure—and the role that structure plays in cellular function, communication, and environmental sensing is an active scientific effort. Previously, we have devised a novel isotopic labeling approach for membrane lipids to enable direct in vivo structural studies of the cell membrane in the Gram-positive bacterium, Bacillus subtilis , using small-angle neutron scattering. This was accomplished through a genetic inhibition of fatty acid (FA) degradation (Δ fadN ) and a chemical inhibition of FA biosynthesis using cerulenin, an irreversible inhibitor of type II fatty acid synthases. Here, we improve upon the previous system by introducing a dCas9/sgRNA- fabF complex that blocks transcription of the essential fabF gene when under xylose induction. This leads to greater sensitivity to cerulenin in the mutant strain (JEBS102) and more robust cell growth when supplementary FAs are introduced to the culture medium. A subtle change in FA uptake is noted when compared to the prior labeling strategy. This is seen in the gas chromatography/mass spectrometry (GC/MS) data as a higher ratio of n 16:0 to a 15:0, and manifests in an apparent increase in the membrane thickness determined via neutron scattering. This represents an improved method of isotopic labeling for the cell membrane of Bacillus subtilis; enabling improved investigations of cellular uptake and utilization of FAs, cell membrane structure and organization as a phenotypic response to metabolic and environmental changes.

59 BASIC BIOLOGICAL SCIENCES↗

Exploring the determinants of organic matter bioavailability through substrate-explicit thermodynamic modeling

Microbial decomposition of organic matter (OM) in river corridors is a major driver of nutrient and energy cycles in natural ecosystems. Recent advances in omics technologies enabled high-throughput generation of molecular data that could be used to inform biogeochemical models. With ultrahigh-resolution OM data becoming more readily available, in particular, the substrate-explicit thermodynamic modeling (SXTM) has emerged as a promising approach due to its ability to predict OM degradation and respiration rates from chemical formulae of compounds. This model implicitly assumes that all detected organic compounds are bioavailable, and that aerobic respiration is driven solely by thermodynamics. Despite promising demonstrations in previous studies, these assumptions may not be universally valid because OM degradation is a complex process governed by multiple factors. To identify key drivers of OM respiration, we performed a comprehensive analysis of diverse river systems using Fourier-transform ion cyclotron resonance mass spectrometry OM data and associated respiration measurements collected by the Worldwide Hydrobiogeochemistry Observation Network for Dynamic River Systems (WHONDRS) consortium. In support of our argument, we found that the incorporation of all compounds detected in the samples into the SXTM resulted in a poor correlation between the predicted and measured respiration rates. The data-model consistency was significantly improved by the selective use of a small subset (i.e., only about 5%) of organic compounds identified using an optimization method. Through a subsequent comparative analysis of the subset of compounds (which we presume as bioavailable) against the full set of compounds, we identified three major traits that potentially determine OM bioavailability, including: (1) thermodynamic favorability of aerobic respiration, (2) the number of C atoms contained in compounds, and (2) carbon/nitrogen (C/N) ratio. We found that all three factors serve as “filters” in that the compounds with undesirable properties in any of these traits are strictly excluded from the bioavailable fraction. This work highlights the importance of accounting for the complex interplay among multiple key traits to increase the predictive power of biogeochemical and ecosystem models.

59 BASIC BIOLOGICAL SCIENCES↗

Controlling circuitry underlies the growth optimization of Saccharomyces cerevisiae

Microbial growth emerges from coordinated synthesis of various cellular components from limited resources. In Saccharomyces cerevisiae, cyclic AMP (cAMP)-mediated signaling is shown to orchestrate cellular metabolism; however, it remains unclear quantitatively how the controlling circuit drives resource partition and subsequently shapes biomass growth. Here we combined experiment with mathematical modeling to dissect the signaling-mediated growth optimization of S. cerevisiae. We showed that, through cAMP-mediated control, the organism achieves maximal or nearly maximal steady-state growth during the utilization of multiple tested substrates as well as under perturbations impairing glucose uptake. However, the optimal cAMP concentration varies across cases, suggesting that different modes of resource allocation are adopted for varied conditions. Under settings with nutrient alterations, S. cerevisiae tunes its cAMP level to dynamically reprogram itself to realize rapid adaptation. Moreover, to achieve growth maximization, cells employ additional regulatory systems such as the GCN2-mediated amino acid control. This study establishes a systematic understanding of global resource allocation in S. cerevisiae, providing insights into quantitative yeast physiology as well as metabolic strain engineering for biotechnological applications.

59 BASIC BIOLOGICAL SCIENCES↗

Data for Controlling Circuitry Underlies the Growth Optimization of Saccharomyces cerevisiae

Microbial growth emerges from coordinated synthesis of various cellular components from limited resources. In Saccharomyces cerevisiae , cyclic AMP (cAMP)-mediated signaling is shown to orchestrate cellular metabolism; however, it remains unclear quantitatively how the controlling circuit drives resource partition and subsequently shapes biomass growth. Here we combined experiment with mathematical modeling to dissect the signaling-mediated growth optimization of S. cerevisiae . We showed that, through cAMP-mediated control, the organism achieves maximal or nearly maximal steady-state growth during the utilization of multiple tested substrates as well as under perturbations impairing glucose uptake. However, the optimal cAMP concentration varies across cases, suggesting that different modes of resource allocation are adopted for varied conditions. Under settings with nutrient alterations, S. cerevisiae tunes its cAMP level to dynamically reprogram itself to realize rapid adaptation. Moreover, to achieve growth maximization, cells employ additional regulatory systems such as the GCN2-mediated amino acid control. This study establishes a systematic understanding of global resource allocation in S. cerevisiae , providing insights into quantitative yeast physiology as well as metabolic strain engineering for biotechnological applications.

Conversion↗

Ocean Biological Pump Sensitivities and Implications for Climate Change Impacts

The ocean is one of the principal reservoirs of CO2, a greenhouse gas, and therefore plays a crucial role in regulating Earth's climate. Currently, the ocean sequesters about a third of anthropogenic CO2 emissions, mitigating the human impact on climate. At the same time, the deeper ocean represents the largest carbon pool in the Earth System and processes that describe the transfer of carbon from the surface of the ocean to depth are intimately linked to the effectiveness of carbon sequestration.The ocean biological pump (OBP), which involves several biogeochemical processes, is a major pathway for transfer of carbon from the surface mixed layer into the ocean interior. About 75 of the carbon vertical gradient is due to the carbon pump with only 25 attributed to the solubility pump. However, the relative importance and role of the two pumps is poorly constrained. OBP is further divided to the organic carbon pump (soft tissue pump) and the carbonate pump, with the former exporting about 10 times more carbon than the latter through processes like remineralization.Major uncertainties about OBP, and hence in the carbon uptake and sequestration, stem from uncertainties in processes involved in OBP such as particulate organicinorganic carbon sinkingsettling, remineralization, microbial degradation of DOC and uptakegrowth rate changes of the ocean biology. The deep ocean is a major sink of atmospheric CO2 in scales of hundreds to thousands of years, but how the export efficiency (i.e. the fraction of total carbon fixation at the surface that is transported at depth) is affected by climate change remains largely undetermined. These processes affect the ocean chemistry (alkalinity, pH, DIC, particulate and dissolved organic carbon) as well as the ecology (biodiversity, functional groups and their interactions) in the ocean. It is important to have a rigorous, quantitative understanding of the uncertainties involved in the observational measurements, the models and the projections of future changes.

particulates↗

Predicting Biological Cleanliness: An Empirical Bayes Approach for Spacecraft Bioburden Accounting

To comply with the international planetary protection policy set forth by the Committee on Space Research and NASA Agency level requirements, spacecraft destined to biologically sensitive planetary bodies have to minimize terrestrial biological contamination. Analysis, testing and inspection are the standard forward verification activities that are used to demonstrate compliance with the biological contamination requirements. For testing of spacecraft surface areas, a swab or wipe sample is collected from surfaces prior to last access and subsequently processed in the lab using NASA Approved Planetary Protection Methods for Culture Based Assays. Raw data resulting from this assay is then statistically treated employing a mathematical paradigm stemming from the 1970’s Viking Lander Project to generate the bioburden density and total microbial bioburden present. This standard approach arbitrarily accounts for error and provides an upper conservative bound as it reports the maximum number of spores estimated to be present on flight hardware surfaces. A bioburden density estimate factors in the following variables: the observed bioburden count, representative volume processed, sampling efficiencies. Notably, to account for error in the approach, a 0 observed count is arbitrarily changed to a count of 1 for each hardware grouping. The data generated by spacecraft bioburden verification campaigns in the past have resulted in <80% of wipes and <90% of swabs containing a bioburden count of 0. As such, having a robust and well documented statistical approach for dealing with the probability of low incident rates is necessary to be able to estimate spacecraft bioburden. Being able to statistically describe the bioburden distribution and associated confidence level is a gamechanger for the development of bioburden allocations during mission design and will allow for tighter management of risk throughout spacecraft build. Thus, Empirical Bayes statistical approach was evaluated to estimate the microbial bioburden on spacecraft to mitigate the aforementioned mathematical concerns and provide a probabilistic bioburden distribution of the flight hardware surface. For application of this approach to performing bioburden calculations, a range of non-informative prior assumptions on hardware surfaces are explored for Bayesian analyses while informative priors using posterior distributions from prior assays are utilized for Empirical Bayes analyses. Several non-informative priors are currently under investigation to assess fitness including use of these priors to serve as a foundation to build off of NASA specification values or a basis of risk to account for unknowns during the integration and testing process. Informative priors under consideration are generated using sampled bioburden values from hardware originating within like processing environments (e.g. vendor cleaning process or similar assembly process), temporal spacecraft status events as a prediction for hardware cleanliness of future samples, and heritage system bioburden actuals to predict allocation for subsequent missions. Informative priors and probabilistic bioburden distributions are then validated using data sets from the Mars Exploration Rover, Mars Science Laboratory, and InSight missions. Using Empirical Bayes approach to generate a probabilistic bioburden distribution as demonstrated through mission use cases provides a valid approach for use in the end-to-end requirements verification process.

97 MATHEMATICS AND COMPUTING↗

Bayesian Framework for Bioburden Density Estimation in Planetary Protection

To comply with the international planetary protection policy set forth by the Committee on Space Research and NASA Agency level requirements, spacecraft destined to biologically sensitive planetary bodies have to minimize terrestrial biological contamination. Analysis, testing and inspection are the standard forward verification activities that are used to demonstrate compliance with the biological contamination requirements. For testing of spacecraft surface areas, a swab or wipe sample is collected from surfaces prior to last access and subsequently processed in the lab using NASA Approved Planetary Protection Methods for Culture Based Assays. Raw data resulting from this assay is then statistically treated employing a mathematical paradigm stemming from the 1970’s Viking Lander Project to generate the bioburden density and total microbial bioburden present. This standard approach arbitrarily accounts for error and provides an upper conservative bound as it reports the maximum number of spores estimated to be present on flight hardware surfaces. A bioburden density estimate factors in the following variables: the observed bioburden count, representative volume processed, sampling efficiencies. Notably, to account for error in the approach, a 0 observed count is arbitrarily changed to a count of 1 for each hardware grouping. The data generated by spacecraft bioburden verification campaigns in the past have resulted in <80% of wipes and <90% of swabs containing a bioburden count of 0. As such, having a robust and well documented statistical approach for dealing with the probability of low incident rates is necessary to be able to estimate spacecraft bioburden. Being able to statistically describe the bioburden distribution and associated confidence level is a gamechanger for the development of bioburden allocations during mission design and will allow for tighter management of risk throughout spacecraft build. Thus, Empirical Bayes statistical approach was evaluated to estimate the microbial bioburden on spacecraft to mitigate the aforementioned mathematical concerns and provide a probabilistic bioburden distribution of the flight hardware surface. For application of this approach to performing bioburden calculations, a range of non-informative prior assumptions on hardware surfaces are explored for Bayesian analyses while informative priors using posterior distributions from prior assays are utilized for Empirical Bayes analyses. Several non-informative priors are currently under investigation to assess fitness including use of these priors to serve as a foundation to build off of NASA specification values or a basis of risk to account for unknowns during the integration and testing process. Informative priors under consideration are generated using sampled bioburden values from hardware originating within like processing environments (e.g. vendor cleaning process or similar assembly process), temporal spacecraft status events as a prediction for hardware cleanliness of future samples, and heritage system bioburden actuals to predict allocation for subsequent missions. Informative priors and probabilistic bioburden distributions are then validated using data sets from the Mars Exploration Rover, Mars Science Laboratory, and InSight missions. Using Empirical Bayes approach to generate a probabilistic bioburden distribution as demonstrated through mission use cases provides a valid approach for use in the end-to-end requirements verification process.

97 - MATHEMATICS AND COMPUTING↗

Engineering Anti-Microbial Peptide Production in Fungi and Bioprocess Development (CRADA 502)

Crop Protection, specifically controlling agricultural diseases and pests, is essential for crop production at every agricultural scale. Biologics hold enormous potential in this equation to decrease use of more toxic pesticides. Currently, the widespread use of biologics in crop protection is severely hampered by the lack of available tools for their mass production. To ensure biologics are regulated under EPA Biopesticide regulations, the following must be true: The biopesticide is a naturally occurring substance or structurally similar and functionally identical to a naturally occurring substance with a history of exposure to humans and the environment demonstrating minimal toxicity. Since this pertains to the marketed product it is also essential that fermentation hosts have a history of exposure and safe use by humans This research focuses on developing a novel platform for the production and delivery of the cghSAMP peptide using the filamentous fungus Aspergillus niger. A. niger is a well-established industrial microorganism, widely recognized for its strong protein secretion capabilities, high-yield production of enzymes, and classification as a generally recognized as safe (GRAS) organism. By genetically modifying A. niger to express the cghSAMP peptide transgene, we aim to leverage the fungus's robust fermentation system to produce a high volume of the antimicrobial agent. This engineered system represents a potential breakthrough in sustainable disease management, offering an environmentally friendly method for localized application or large-scale production of a biopesticide designed to mitigate the effects of citrus greening disease. The main objectives of this project were: 1. Developing a high-yield and cost-efficient bioprocess to produce an AMP in a current ABF host or readily onboarded hosts. 2. Scale up the process in stirred tank reactors and produce at least 100 grams of AMP at 1- 5g/L

60 APPLIED LIFE SCIENCES↗

UV Shielding of Bacillus pumilus SAFR-032 Endospores by Martian Regolith Simulants

As exploration of the solar system advances with life detection missions on the horizon, the concern for planetary protection has grown considerably. When attempting to detect extraterrestrial life, the likelihood of false positives from terrestrial contamination must be minimized. The Exposing Microorganisms in the Stratosphere (E-MIST) balloon project aims to evaluate whether resilient terrestrial bacteria can survive stressors in a Mars-like environment. This is accomplished by sending Bacillus pumilus SAFR-032, an endospore-forming bacterial isolate from a spacecraft assembly facility, to the Earth's middle stratosphere (30-38 kilometers), where low temperature and pressure and high radiation and dryness conditions are similar to the surface of Mars. Previous ground and flight tests showed that the vast majority of SAFR-032 spores (99.99 percent) were inactivated by direct sunlight due to ultraviolet (UV) radiation. This observation led us to explore the role of dust shielding in changing microbial survivorship outcomes. To determine the dust particle distributions and density for potentially shielding microbes from UV radiation, samples of a Martian dust simulant were mixed with SAFR-032 spores. The dry heat sterilized simulant used was JSC MARS-1, weathered volcanic ash from Hawaii that displays many chemical and physical properties similar to the Martian soil as characterized by the Viking Lander 1, including reflectance spectrum, chemical composition, mineralogy, grain size, specific gravity, and magnetic properties. First, scanning electron microscopy was undertaken to visualize the aggregation of the spores with dust particles (i.e., shading effects), and samples of varying dust concentrations were subsequently irradiated with UVC light to test survivorship outcomes. After a relationship between dust concentration and spore survivorship was determined, a solar simulator capable of irradiating samples with a fuller UV spectrum (less than 280-400 nanometers) was used to perform a more robust middle stratosphere simulation. Taken together, we will use results from the ground-based irradiation studies to feed into experimental designs for the next E-MIST ultra-long duration polar balloon flight launched by NASA.

Stratosphere↗

Advanced microbial check valve development

A flight certified assembly identified as a Microbial Check Valve (MCV) was developed and tested. The MCV is a canister packed with an iodinated anionic exchange resin. The device is used to destroy organisms in a water stream as the water passes through the device. The device is equally effective for fluid flow in either direction and its primary method of organism removal is killing rather than filtering. The MCV was successfully developed for the space shuttle to: disinfect fuel cell water; and prevent back contamination of the stored potable water supply. One version of the device consists of a high residual iodinated resin bed that imparts approximately 2 ppm of iodine to the fuel cell water as it flows to the potable water tanks. A second version of the device consists of a low residual iodinated resin bed. One of these low residual beds is located at each use port in the potable water system for the dual purpose of removing some iodine from the potable water as it is dispensed and also to prevent back contamination of the potable supply.

Colombo, G. V.↗

An Improved UAV-Based Fog Collector for Local-Scale Aerobiological Sampling

The use of unmanned air vehicles (UAVs) to passively collect fog for microbiological analysis is being investigated at NASA Ames, contributing to our understanding of the dynamics and limits of the biosphere. Current improvements to a preliminary payload design include physically modifying the mesh of the passive impactor collection unit (PICU) to increase fog-capture efficiency and a quick- mounting system to improve sample turnaround and contamination control. Increasing the water-capture efficiency of the PICU involved changing the surface roughness of the polypropylene mesh by abrasive sanding. Four scale models of the PICU were created, with three of those models sanded with 80-, 150-, and 240-grit sand paper for a minimum of thirty seconds on each side. Each mesh was individually tested in a fog-simulation chamber to measure collection efficiency. Tests showed that sanding the mesh will increase the water-capture efficiency by at least 3-fold for the 150- and 240-grit. Next steps are to determine the scaling laws and minimum amount of exposure time as a function of the fog density, to acquire enough microorganisms for analysis; a light-weight ground module for the PICU has been developed to allow rapid, easy field testing. Developing a mounting system that will ensure sterile deployment, as well as prompt changes of PICUs, for collection of fog water, utilized computational fluid dynamics (CFD) for the UAV to ensure that the location of the impactors will not be contaminated by particulates in the UAV downwash. The original design allowed the impactors to remain within sterile packaging until mounted on the UAV but did not address the concerns of contamination from aerodynamic ground effects and particulate lofting during takeoff. Design for a raised launch pad would allow for the sterile PICU to be attached to UAV and launched from outside the region of projected particulate lofting. California coastal sites have been scouted for appropriate testing locations. The improvements to the PICU design and operations will allow for multiple sampling points within a single fog event. The returned fog samples will undergo ion chromatography, qPCR, ATP quantification, and other tests to characterize microbial species and relevant biochemistry.

Wong, Amber↗

Effects of hurricane disturbance and increased temperature on carbon cycling and storage of a Puerto Rican forest: a mechanistic investigation of above- and belowground processes (Final Technical Report)

The overall goal of the Tropical Responses for Altered Climate Experiment (TRACE) is to assess the effects of increasing temperature on tropical plant and soil carbon fluxes and storage as the forest recovers from major hurricane disturbance that occurred in September 2017. Ultimately, we aimed to reduce uncertainty and increase confidence with which tropical forests are represented in Earth System models to make more accurate global forecasts of future climate. We focused on both above- and belowground processes and explored temperature controls over critical aspects of carbon and nutrient cycling for tropical plants, soil, and microbes. TRACE is located in a wet tropical forest in the Luquillo Experimental Forest close to the USDA Forest Service Sabana Field Research Station in Luquillo, Puerto Rico. The warming treatment consists of six 4.7 m diameter plots. Three of the plots receive infrared warming and three have the same infrastructure but are not warmed (using ‘dummy’ heaters). Each plot was monitored from 2018-2023 to investigate two major questions: 1) Are there legacy effects of prior warming on forest recovery following hurricane disturbance? 2) Will the trajectory of forest recovery following disturbance be affected by warmer temperatures? Concurrent soil incubation experiments were conducted to enable more controlled mechanistic investigations of temperature response on microbial function. In sum, our goal was to use this novel climate manipulation experiment (the only of its kind in any tropical forest) and once-in-a-lifetime chance to assess how temperature and hurricane disturbance interact to affect coupled biogeochemical cycling in a tropical forest.

54 ENVIRONMENTAL SCIENCES↗