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NASA NTRS2011

Legacy of Environmental Research During the Space Shuttle Program

The Space Shuttle Program provided many opportunities to study the role of spaceflight on human life for over the last 30 years and represents the longest and largest U.S. human spaceflight program. Risks to crewmembers were included in the research areas of nutrition, microbiology, toxicology, radiation, and sleep quality. To better understand the Shuttle environment, Crew Health Care System was developed. As part of this system, the Environmental Health Subsystem was developed to monitor the atmosphere for gaseous contaminants and microbial contamination levels and to monitor water quality and radiation. This program expended a great deal of effort in studying and mitigating risks related to contaminations due to food, water, air, surfaces, crewmembers, and payloads including those with animals. As the Shuttle had limited stowage space and food selection, the development of nutritional requirements for crewmembers was imperative. As the Shuttle was a reusable vehicle, microbial contamination was of great concern. The development of monitoring instruments that could withstand the space environment took several years and many variations to come up with a suitable instrument. Research with space radiation provided an improved understanding of the various sources of ionizing radiation and the development of monitoring instrumentation for space weather and the human exposure within the orbiter's cabin. Space toxicology matured to include the management of offgassing products that could pollute the crewmembers air quality. The Shuttle Program implemented a 5-level toxicity rating system and developed new monitoring instrumentation to detect toxic compounds. The environment of space caused circadian desynchrony, sleep deficiency, and fatigue leading to much research and major emphasis on countermeasures. Outcomes of the research in these areas were countermeasures, operational protocols, and hardware. Learning Objectives: This symposium will provide an overview of the major environmental lessons learned and the development of countermeasures, monitoring hardware, and procedures.

Lane, Helen W.↗

NASA NTRSDate not supplied

Strategies for Determining Safety of Fermented Foods Produced in Space

The BioNutrients experiments examine the feasibility of using microbially produced foods – yogurt, kefir, fermented food products– to supplement the current prepackaged food system to provide freshly synthesized nutrients and genetically engineered protein therapeutics to support crew health for sustainable and long duration crewed exploration. Safety of the products must be established as direct consumption of such products is a risk due to potential contamination with pathogenic organisms. Identifying pathogen contamination is particularly challenging as the fermenting cultures can mask the presence of contaminating organisms. Here we consider current methods of detecting coliforms, aerobic colonies, non-lactic acid bacteria, molds (yeasts), and specific pathogens including Staphylococcus aureus and salmonella. We also consider alternate and additional testing including sequencing and quantitative PCR. We investigate the use of the E-Nose developed by NASA Ames Research Center. This portable device senses volatile organic compounds in real time and can be trained to recognize “good” uncontaminated products from contaminated ones as a means of predicting food safety. Finally, we consider methods of in-flight pasteurization to increase safety and decrease overall microbial load of these products. This presentation will summarize testing status and outline the strategy that will be employed in the BioNutrients-3 ISS fermented foods experiment.

Natalie Ball↗

NASA NTRSDate not supplied

Strategies for Determining Safety of Fermented Foods Produced in Space

Food↗

DOE OSTI2025

Phosphate-modulated transformation of Sb(V)-bearing ferrihydrite under microbial iron- and sulfate-reducing conditions

Antimony (Sb) is a toxic metalloid that poses environmental risks in terrestrial and aquatic systems. The fate of the Sb(V) oxyanion, Sb(OH) 6 − , is governed by complex biogeochemical processes, including immobilization by ferric (Fe(III)) oxides, reduction by sulfide, and the less-explored competitive adsorption with other anions such as phosphate (PO 4 3− ). Here, this study investigates the interplay between such mechanisms in controlling the behavior of Sb(V) associated with ferrihydrite (Fh) under Fe(III)- and sulfate-reducing conditions, with a particular emphasis on the role of phosphate in influencing Sb(V) mobility and transformation. Anoxic reactors that contained Sb(V)-coprecipitated Fh, varying PO 4 3− concentrations (0, 0.2, and 2 mM), and sulfate, were inoculated with a microbial community sourced from Sb-contaminated soil. Additionally, abiotic reactors with either Sb(V)-adsorbed or Sb(V)-coprecipitated Fh and different PO 4 3− loadings (0–100 mM) were created to investigate the competitive adsorption mechanisms in the absence of microbial activity. Results from the abiotic reactors suggest that Sb(V) is likely incorporated into the Fh structure, with only minor amounts remaining surface-bound and extractable by PO 4 3− . In the biotic reactors, microbial Fe(III) and sulfate reduction were more extensive in the presence of PO 4 3− . At 0.2 mM PO 4 3− , microbial activity transformed Fh into siderite and led to the complete reduction of Sb(V) to Sb(III) as stibnite (Sb 2 S 3 ). At 2 mM PO 4 3− , the greater coverage by PO 4 3− stabilized Fh and decreased the extent of both Fe(III) and Sb(V) reduction, and shifted the reduced products to mackinawite and Sb(III) adsorbed onto Fh, in addition to stibnite formation. This study demonstrates that while PO 4 3− may not directly compete with Sb(V) for sorption sites, it can influence Sb mobility in Fe(III)- and sulfate-reducing environments by enhancing microbial activity and altering the mineralization pathways.

Microbial Fe(III) and sulfate reduction↗

DOE OSTI2025

Upstream considerations for gas fermentation processes

Gas fermentation enables the production of fuels, chemicals, and foods from gaseous carbon sources and could serve as a technology for valorizing carbon that may otherwise be emitted to the atmosphere. In this review, we focus on upstream feedstock considerations: the supply of carbon and the supply of electrical power. Electrical power serves a dual role, providing both process energy and biochemical redox potential (via hydrogen or reduced intermediates). We define gas fermentation as bioprocesses involving gaseous feedstocks metabolized by microbes, distinct from microbial electrosynthesis. Trends in CO 2 point sources and low-carbon electricity systems are analyzed, highlighting opportunities and challenges for future deployment. This review synthesizes current knowledge and identifies key R&D priorities for process integration at industrial scale.

09 BIOMASS FUELS↗

DOE OSTI2021

Microbial Taxonomy Run Amok

DNA sequencing has led to an explosion in discovery of microbial phylogenetic novelty, especially that represented by uncultivated taxa, to which the traditional system of prokaryotic taxonomy has not adapted. A lack of expansion of the International Code of Nomenclature of Prokaryotes (ICNP, 'the Code') to effectively capture this information has created a 'wild west' situation where names are published or appear in popular reference databases without further verification or validation. The rapid propagation of variant and questionable naming methods has led to widespread confusion and undermines prior accomplishments. Furthermore, we exemplify inconsistencies that have arisen from this practice and endanger the interoperability of scientific findings. The immediate solution to this problem is to develop and adopt universal best practices that are accepted by expert researchers, major publishers, the International Committee on Systematics of Prokaryotes (ICSP), and international microbiological societies.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Cyanobacteria from marine oxygen-deficient zones encode both form I and form II Rubiscos

Cyanobacteria are highly abundant in the marine photic zone and primary drivers of the conversion of inorganic carbon into biomass. To date, all studied cyanobacterial lineages encode carbon fixation machinery relying upon form I Rubiscos within a CO 2 -concentrating carboxysome. Here, we report that the uncultivated anoxic marine zone (AMZ) IB lineage ofProchlorococcusfrom pelagic oxygen-deficient zones (ODZs) harbors both form I and form II Rubiscos, the latter of which are typically noncarboxysomal and possess biochemical properties tuned toward low-oxygen environments. We demonstrate that these cyanobacterial form II enzymes are functional in vitro and were likely acquired from proteobacteria. Metagenomic analysis reveals that AMZ IB are essentially restricted to ODZs in the Eastern Pacific, suggesting that form II acquisition may confer an advantage under low-O 2 conditions. AMZ IB populations express both forms of Rubisco in situ, with the highest form II expression at depths where oxygen and light are low, possibly as a mechanism to increase the efficiency of photoautotrophy under energy limitation. Our findings expand the diversity of carbon fixation configurations in the microbial world and may have implications for carbon sequestration in natural and engineered systems.

Science & Technology - Other Topics↗

DOE OSTI2021

Plant-microbe interactions: from genes to ecosystems using Populus as a model system

Plant-microbe symbioses span a continuum from pathogenic to mutualistic with functional consequences for both organisms in the symbiosis. In order to increase sustainable food and fuel production in the future, it is imperative that we harness these symbioses. The tree genus Populus is an excellent model system for studies examining plant-microbe interactions due to the wealth of genomic information available and the molecular tools that have been developed to manipulate Populus-microbe symbioses. In this review, we highlight how Populus can serve as a model system to explore plant-microbe interactions. Specifically, highlighting research linking Populus-microbe interactions from the gene to the ecosystem level. We explore why Populus is an excellent model for perennial plant systems, the molecular underpinnings of Populus-microbe interactions, how host genetics influence microbial community composition, and how microbial communities vary at fine spatial scales and between Populus species. Further, we explore how the patterns of the microbiome may affect ecosystem level functions in managed and natural ecosystems. Understanding and manipulating these interactions in Populus has the potential to improve plant health and impact ecosystem sustainability and processes as Populus trees function as foundational species in many natural ecosystems and are also deployed in managed ecosystems for various agroforestry applications.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2022

Thermodynamic Constraints on Electromicrobial Protein Production

Global consumption of protein is projected to double by the middle of the 21st century. However, protein production is one of the most energy intensive and environmentally damaging parts of the food supply system today. Electromicrobial production technologies that combine renewable electricity and CO 2 -fixing microbial metabolism could dramatically increase the energy efficiency of commodity chemical production. Here we present a molecular-scale model that sets an upper limit on the performance of any organism performing electromicrobial protein production. We show that engineered microbes that fix CO 2 and N 2 using reducing equivalents produced by H 2 -oxidation or extracellular electron uptake could produce amino acids with energy inputs as low as 64 MJ kg −1 , approximately one order of magnitude higher than any previous estimate of the efficiency of electromicrobial protein production. This work provides a roadmap for development of engineered microbes that could significantly expand access to proteins produced with a low environmental footprint.

60 APPLIED LIFE SCIENCES↗

NASA NTRS1970

Biological handbook for engineers

Microbiological background information is compiled in handbook for engineers and scientists working on bio-related projects. It is intended as aid in - /1/ evaluating effects of engineering procedures on microbial life, /2/ determining effects of decontamination and sterilization on performance of overall systems, and /3/ understanding language of microbiologists.

Source record↗

NASA NTRS1973

Cuvette loading and unloading procedures

The cuvettes for the flight hardware used in the Microbial Response to Space Environment Experiment (M191) were loaded with the biological test systems according to the methods described. After the flight, the experiment package was returned and dismantled. Then, the individual cuvettes were removed from the hardware and unloaded according to the procedures described.

Carmichael, C.↗

NASA NTRS1990

Spacelab 3 mission

Spacelab-3 (SL-3) was the first microgravity mission of extended duration involving crew interaction with animal experiments. This interaction involved sharing the Spacelab environmental system, changing animal food, and changing animal waste trays by the crew. Extensive microbial testing was conducted on the animal specimens and crew and on their ground and flight facilities during all phases of the mission to determine the potential for cross contamination. Macroparticulate sampling was attempted but was unsuccessful due to the unforseen particulate contamination occurring during the flight. Particulate debris of varying size (250 micron to several inches) and composition was recovered post flight from the Spacelab floor, end cones, overhead areas, avionics fan filter, cabin fan filters, tunnel adaptor, and from the crew module. These data are discussed along with solutions, which were implemented, for particulate and microbial containment for future flight facilities.

Dalton, Bonnie P.↗

NASA NTRS1992

Aquatic biofilms and their responses to disinfection and invading species

The control of microbial contamination is a primary concern in the development of a water reclamation system for long-duration manned space flights. This paper describes bench-scale experiments, using both static and recycling water systems, investigating the interaction of bacterial species in the development of a biofilm and their response to the introduction of a disinfectant or of additional species. The results showed that iodine concentrations as high as 15 to 20 mg/l I2 are necessary to completely disinfect a stable biofilm. When S. aueus and E. coli were introduced into a system containing natural mixed culture biofilms, their colonization in the biofilms increased their survival time, from 3 to 5 days as unattached cells to over 60 days when protected in the biofilms. While iodine concentrations of 0.5 to 1 mg/l were enough to eliminate these organisms from the bulk water, concentrations higher than 4.0 mg/l were necessary to completely eliminate these organisms from the biofilm.

Smithers, G. A.↗

NASA NTRS1998

Beating Back Bacteria

Under a NASA-Johnson Space Center contract, Umpqua Research developed the MCV (Trademark) (Microbial Check Valve) which uses iodinated ion exchange resin used for water purification systems aboard space missions. Using this resin, MRLB International, Inc., developed and commercialized the Dentapure purification cartridge used by dentists nationwide.

Source record↗

NASA NTRS2010

Automated, Miniaturized Instrument for Measuring Gene Expression in Space

To facilitate astrobiological studies on the survival and adaptation of microorganisms and mixed microbial cultures to space environment, we have been developing a fully automated, miniaturized system for measuring their gene expression on small spacecraft. This low-cost, multi-purpose instrument represents a major scientific and technological advancement in our ability to study the impact of the space environment on biological systems by providing data on cellular metabolism and regulation orders of magnitude richer than what is currently available. The system supports growth of the organism, lyse it to release the expressed RNA, label the RNA, read the expression levels of a large number of genes by microarray analysis of labeled RNA and transmit the measurements to Earth. To measure gene expression we use microarray technology developed by CombiMatrix, which is based on electrochemical reactions on arrays of electrodes on a semiconductor substrate. Since the electrical integrity of the microarray remains intact after probe synthesis, the circuitry can be employed to sense nucleic acid binding at each electrode. CombiMatrix arrays can be sectored to allow multiple samples per chip. In addition, a single array can be used for several assays. The array has been integrated into an automated microfluidic cartridge that uses flexible reagent blisters and pinch pumping to move liquid reagents between chambers. The proposed instrument will help to understand adaptation of terrestrial life to conditions beyond the planet of origin, identify deleterious effects of the space environment, develop effective countermeasures against these effects, and test our ability to sustain and grow in space organisms that can be used for life support and in situ resource utilization during long-duration space exploration. The instrument is suitable for small satellite platforms, which provide frequent, low cost access to space. It can be also used on any other platform in space, including the ISS. It can be replicated and used with only small modifications in multiple biological experiments with a broad range of goals in mind.

Pohorille, A.↗

NASA NTRSDate not supplied

Operation of the Urine Collection and Pretreatment System in the ISS US Segment Waste & Hygiene Compartment (WHC)

The US Segment Waste & Hygiene Compartment (WHC)provides the capability to collect crew urine for subsequent delivery to the Urine Processor Assembly (UPA). Urine collection in the WHC is accomplished with the urine collection and pretreatment system designed by RSC “Energia” and NIICHIMMASH and integrated into the WHC by Boeing and NASA. The WHC was installed in the US Laboratory Module in November 2008 and later transferred to the Node 3 in February 2010.As of February 1st 2021, the WHC has collected 25855 liters of pretreated urine. The system collects urine from astronauts, adds required pretreatment chemicals to maintain chemical and microbial control of the urine, and delivers the pretreated urine to the Urine Processor Assembly. This paper will summarize the twelve years of operation of the WHC on ISS, significant anomalies, and lessons learned from its operation.

D Layne Carter↗

DOE OSTI2020

Increased microbial growth, biomass, and turnover drive soil organic carbon accumulation at higher plant diversity

Species-rich plant communities have been shown to be more productive and to exhibit increased long-term soil organic carbon (SOC) storage. Soil microorganisms are central to the conversion of plant organic matter into SOC, yet the relationship between plant diversity, soil microbial growth, turnover as well as carbon use efficiency (CUE) and SOC accumulation is unknown. As heterotrophic soil microbes are primarily carbon limited, it is important to understand how they respond to increased plant-derived carbon inputs at higher plant species richness (PSR). We used the long-term grassland biodiversity experiment in Jena, Germany, to examine how microbial physiology responds to changes in plant diversity and how this affects SOC content. The Jena Experiment considers different numbers of species (1–60), functional groups (1–4) as well as functional identity (small herbs, tall herbs, grasses, and legumes). We found that PSR accelerated microbial growth and turnover and increased microbial biomass and necromass. PSR also accelerated microbial respiration, but this effect was less strong than for microbial growth. In contrast, PSR did not affect microbial CUE or biomass-specific respiration. Structural equation models revealed that PSR had direct positive effects on root biomass, and thereby on microbial growth and microbial biomass carbon. Finally, PSR increased SOC content via its positive influence on microbial biomass carbon. We suggest that PSR favors faster rates of microbial growth and turnover, likely due to greater plant productivity, resulting in higher amounts of microbial biomass and necromass that translate into the observed increase in SOC. We thus identify the microbial mechanism linking species-rich plant communities to a carbon cycle process of importance to Earth's climate system.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRSDate not supplied

SERFE Water Quality Results

NASA has been developing a new spacesuit, called the Exploration Extravehicular Mobility Unit (xEMU) for over a decade. This spacesuit is underdevelopment to support missions to the International Space Station (ISS) and also to the Moon. Improvements in the life and robustness of the Portable Life Support System (PLSS) has been a major objective of these efforts. Water quality, and how that impacts the performance of the life support system hardware, are areas where improvements could greatly help to achieve these goals. The Suit Water Membrane Evaporator (SWME) was chosen as the technology to provide cooling to the xEMU and has undergone several iterations of development over this period. An ISS flight experiment centered around the SWME and other thermal loop technologies was developed and has been under test in an ISS EXpedite PRocessing of Experiments to the Space Station (EXPRESS) rack since November of 2020. In addition to the SWME, The SWME EXPRESS Rack Flight Experiment (SERFE) contains several technology demonstrations from the xEMU project and has been demonstrating their performance in micro-gravity and over an extended duration. This paper will summarize the results of the water quality analysis performed to date throughout the SERFE testing program. Special care was taken to select materials, process hardware, and control the quality of the feedwater used in SERFE testing so that it would be representative of a thermal control system that could be used on a spacesuit for a lunar mission. In addition, the testing schedule, including periods of quiescence in between tests, was also selected to simulate a real mission profile in ways that would be critical to water quality. Results include data and analysis on microbial activity, biocidal iodine, metals, organic carbon, and several other customary indicators of water quality in an operational system. Testing spans over a year and a half of wetted life of the SERFE Flight Unit and the SERFE Ground Unit.

Colin Campbell↗