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At least 613 records · Page 34

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NASA NTRS2015

Analysis of Process Gases and Trace Contaminants in Membrane-Aerated Gaseous Effluent Streams.

In membrane-aerated biofilm reactors (MABRs), hollow fibers are used to supply oxygen to the biofilms and bulk fluid. A pressure and concentration gradient between the inner volume of the fibers and the reactor reservoir drives oxygen mass transport across the fibers toward the bulk solution, providing the fiber-adhered biofilm with oxygen. Conversely, bacterial metabolic gases from the bulk liquid, as well as from the biofilm, move opposite to the flow of oxygen, entering the hollow fiber and out of the reactor. Metabolic gases are excellent indicators of biofilm vitality, and can aid in microbial identification. Certain gases can be indicative of system perturbations and control anomalies, or potentially unwanted biological processes occurring within the reactor. In confined environments, such as those found during spaceflight, it is important to understand what compounds are being stripped from the reactor and potentially released into the crew cabin to determine the appropriateness or the requirement for additional mitigation factors. Reactor effluent gas analysis focused on samples provided from Kennedy Space Center's sub-scale MABRs, as well as Johnson Space Center's full-scale MABRs, using infrared spectroscopy and gas chromatography techniques. Process gases, such as carbon dioxide, oxygen, nitrogen, nitrogen dioxide, and nitrous oxide, were quantified to monitor reactor operations. Solid Phase Microextraction (SPME) GC-MS analysis was used to identify trace volatile compounds. Compounds of interest were subsequently quantified. Reactor supply air was examined to establish target compound baseline concentrations. Concentration levels were compared to average ISS concentration values and/or Spacecraft Maximum Allowable Concentration (SMAC) levels where appropriate. Based on a review of to-date results, current trace contaminant control systems (TCCS) currently on board the ISS should be able to handle the added load from bioreactor systems without the need for secondary mitigation.

Gaseous Effluent Streams↗

NASA NTRS2016

Results of the Alternative Water Processor Test, A Novel Technology for Exploration Wastewater Remediation

Biologically-based water recovery systems are a regenerative, low energy alternative to physiochemical processes to reclaim water from wastewater. This report summarizes the results of the Alternative Water Processor (AWP) Integrated Test, conducted from June 2013 until April 2014. The system was comprised of four (4) membrane aerated bioreactors (MABRs) to remove carbon and nitrogen from an exploration mission wastewater and a coupled forward and reverse osmosis system to remove large organic and inorganic salts from the biological system effluent. The system exceeded the overall objectives of the test by recovering 90% of the influent wastewater processed into a near potable state and a 64% reduction of consumables from the current state of the art water recovery system on the International Space Station (ISS). However, the biological system fell short of its test goals, failing to remove 75% and 90% of the influent ammonium and organic carbon, respectively. Despite not meeting its test goals, the BWP demonstrated the feasibility of an attached-growth biological system for simultaneous nitrification and denitrification, an innovative, volume- and consumable-saving design that does not require toxic pretreatment.

Meyer, Caitlin E.↗

NASA NTRS2015

Plant Biomass Leaching for Nutrient Recovery in Closed Loop Systems Project

Plants will be important for food and O2 production during long term human habitation in space. Recycling of nutrients (e.g., from waste materials) could reduce the resupply costs of fertilizers for growing these plants. Work at NASA's Kennedy Space Center has shown that ion exchange resins can extract fertilizer (plant essential nutrients) from human waste water, after which the residual brine could be treated with electrodialysis to recover more water and produce high value chemicals (e.g., acids and bases). In habitats with significant plant production, inedible biomass becomes a major source of solid waste. To "close the loop" we also need to recover useful nutrients and fertilizer from inedible biomass. We are investigating different approaches to retrieve nutrients from inedible plant biomass, including physical leaching with water, processing the biomass in bioreactors, changing the pH of leaching processing, and/or conducting multiple leaches of biomass residues.

Technology Portfolio System↗

NASA NTRS2016

FOST 2 Upgrade with Hollow-Fiber CTA FO Module and Generation of Osmotic Agent for Microorganism Growth Studies

FOST 2 is an integrated membrane system that incorporates a forward osmosis subsystem and a reverse osmosis subsystem working in series. It has been designed as a post treatment system to process the effluent from the Membrane Aerated Biological Reactor developed at NASA Johnson Space Center and Texas Tech University. Its function is to remove dissolved solids residual such as ammonia and suspended solids, as well as to provide a physical barrier to microbial and viral contamination. A tubular CTA membrane module from HTI and a flat-sheet lipid-base membrane module from Porifera were integrated and tested on FOST 2 in the past, using both a bioreactor's effluent and greywater as the feed solution. This paper documents the performance of FOST 2 after its upgrade with a hollow-fiber CTA membrane module from Toyobo, treating real black-water to generate the osmotic agent solution necessary to conduct growth studies of genetically engineered microorganism for the Synthetic Biological Membrane project.

Water treatment↗

NASA NTRS2016

Development of the Two Phase Flow Separator Experiment for a Reduced Gravity Aircraft Flight

The recent hardware development and testing of a reduced gravity aircraft flight experiment has provided valuable insights for the future design of the Two Phase Flow Separator Experiment (TPFSE). The TPFSE is scheduled to fly within the Fluids Integration Rack (FIR) aboard the International Space Station (ISS) in 2020. The TPFSE studies the operational limits of gas and liquid separation of passive cyclonic separators. A passive cyclonic separator utilizes only the inertia of the incoming flow to accomplish the liquid-gas separation. Efficient phase separation is critical for environmental control and life support systems, such as recovery of clean water from bioreactors, for long duration human spaceflight missions. The final low gravity aircraft flight took place in December 2015 aboard NASA's C9 airplane.

fluid mechanics↗

NASA NTRS2016

Results of the Alternative Water Processor Test, A Novel Technology for Exploration Wastewater Remediation

Biologically-based water recovery systems are a regenerative, low energy alternative to physiochemical processes to reclaim water from wastewater. This paper summarizes the results of the Alternative Water Processor (AWP) test conducted over one year. The AWP recovered 90% of water from four crewmembers using (4) membrane aerated bioreactors (MABRs) to remove carbon and nitrogen from an exploration mission wastewater, including urine, hygiene, laundry and humidity condensate. Downstream, a coupled forward and reverse osmosis system removed large organics and inorganic salts from the biological system effluent. The system exceeded the overall objectives of the test by recovering 90% of the influent wastewater processed and a 29% reduction of consumables from the current state of the art water recovery system on the International Space Station (ISS). However the biological system fell short of its test goals, failing to remove 75% and 90% of the influent ammonium and organic carbon, respectively. Despite not meeting its test goals, the BWP demonstrated the feasibility of an attached-growth biological system for simultaneous nitrification and denitrification, an innovative, volume and consumable-saving design that doesn't require toxic pretreatment. This paper will explain the reasons for this and will discuss steps to optimize each subsystem to increase effluent quality from the MABRs and the FOST to advance the system.

Leticia Vega↗

NASA NTRS2018

Closed Loop Water Purification System Utilizing an Algae Membrane Photobioreactor for the International Space Station

The International Space Station (ISS) currently operates on an open-loop water purification and distribution system. This system is only 75% efficient with a 25% water loss that must be resupplied on a quarterly basis through cargo resupply missions (Pickett, M.). This system is not feasible for long-duration spaceflight missions. A closed-loop water purification system aboard the ISS can benefit water reclamation and become the water purification system for long-term space flight. The primary sub-systems integrated within the proposed closed loop system include an anaerobic digestion membrane bioreactor and an algae membrane photobioreactor. The processes mimics the abilities of natural biological systems naturally. The water that flows from the algal membrane photobioreactor is rich in nutrients providing a feed source for plants aboard the ISS or can be further processed for potable water.

photobioreactor↗

NASA NTRS2019

Planetary Water Recycling Systems Trade Study

Long-term planetary space missions present new and unique challenges in life-support systems. Water constitutes the majority of the mass required to sustain human life in space and it follows that efficient water recycling has the potential to lower mission costs. The effect of partial gravity in planetary missions mean that terrestrial systems could be applicable. This trade study evaluates terrestrial and NASA developed water recycling technologies on the basis of applicability as a planetary base water recycling systems. Various bioreactors, membrane reactors, filtration, and district water reclamation systems are investigated and rated based on several standardized parameters. A customer-oriented Quality Function Deployment (QFD) is utilized to analyze the ratings of the technologies for the tasks required. The trade study aims to rank the various systems based on their Equivalent System Mass (ESM), Technology Readiness Level (TRL), scalability, crew time, and overall logistics requirements, among others. The results of the study can serve as a basis for future inquiries and studies by NASA and other interested parties. The results of this study provide a down selection from 24 systems to 5 systems that trade very close to each other. The results provide a context and justification for a future comparative hardware test program to determine which of these systems offer the best solution.

Flynn, Michael↗

NASA NTRSDate not supplied

BioNutrients: Microbial on-demand production of short shelf-life micronutrients in space

Long duration deep space missions pose a significant challenge when considering adequate nutrition for human spaceflight. Many micronutrients supplied in pre-packaged foods for low Earth orbit flights have a short shelf life and will degrade over multi-year missions. One strategy to mitigate the risk of micronutrient malnutrition is to grow fresh foods to supplement a pre-packaged diet. The BioNutrients project aims to develop systems for growing edible microorganisms that are genetically engineered to produce short shelf life micronutrients. BioNutrients-1 (BN-1) is a five-year spaceflight project currently underway on the International Space Station (ISS) that tests long-term storage and production of two carotenoids, β-carotene and zeaxanthin, in genetically engineered yeast strains. BN-1 on-orbit and ground control activities indicate that the nutrient production system will be stable for at least two years in ambient storage conditions. BN-1 is also testing the effects of long-term ISS storage on viability and gene expression using “stasis packs” containing a series of microorganisms useful in food and biomanufacturing processes. Stasis pack analysis is distinguishing microorganisms with full viability after two years of storage from other species that lose viability rapidly. In parallel, BioNutrients-2 (BN-2) is being developed as a six-month flight experiment that will expand the range of foods produced on ISS to include yogurt, kefir, and probiotic yeast. BN-2 will improve upon hardware design by transitioning from a hard-shell polycarbonate bioreactor to a flexible fluorinated ethylene propylene (FEP) bag. This change reduces the mass and volume required to store production packs. The long-term goal of BioNutrients is to develop the capability for rapid and safe delivery of multiple micronutrients in single-use production packs. We expect synthetic biology approaches similar to BioNutrients will play an important role for astronaut health in future NASA missions to the Moon and Mars.

Synthetic Biology↗

NASA NTRSDate not supplied

Tracking Metabolic Changes in Microbial Culture using Redox Measurements

During long-term space missions, microbial cultures accumulate the effects of low-dose radiation, microgravity, and other factors; altered growth and metabolic activity may occur before viability effects. This could affect functionality of bioreactors or other bio-enabled mission systems, as well as shed light on human health. Spaceflight microbiology studies beyond the low Earth orbit exposure afforded by the ISS have been limited. Nanosatellites offer an increasingly popular alternative for deep space missions. However, the communications delay requires biofluidic automation of a pre-defined experimental protocol, and the lack of sample return (reliance on sensors in flight) can significantly limit feasible investigations. Previous biological CubeSats (PharmaSat, O/OREOS, EcAMSat) have used alamarBlue, an off-the-shelf formulation of the redox indicator dye resazurin, to track metabolic activity, as will BioSentinel, the upcoming interplanetary microbiology experiment. A series of ground experiments (see abstracts by Liddell, Santa Maria, and A. Kim) were conducted using a microbial culture system outfitted with an electrochemical sensor array (electrical conductivity, pH, oxidation-reduction potential, and dissolved oxygen) with alamarBlue and the same strain of Saccharomyces cerevisiae as BioSentinel. By improving mapping of measured changes in alamarBlue kinetics to physicochemical changes, and ultimately to biological alterations such as shifted metabolic pathways, this work supplements data analyses from past missions and planning for future missions using alamarBlue to characterize space radiation effects. Initial results indicate that alamarBlue acts like a redox buffer; its presence significantly changes redox kinetics in otherwise identical cultures. The initial color change (blue resazurin reduced to red/pink resorufin) appears as a redox plateau. A second plateau, likely corresponding to the second color transition (resorufin to the colorless hydroresorufin), occurs at a lower redox value. The relationship to carbon source exhaustion, dissolved oxygen depletion, cell death, and measured redox potential is complex and still under study.

Tracking↗

NASA NTRSDate not supplied

Investigation into Stimulated Microgravity Techniques used to Study Biofilm Growth

Bacterial growth in liquid media in microgravity conditions is not well understood. Trends such as a shortened lag phase, longer log phase, slower growth rate, and a higher final population concentration have been noted but the underlying cause remains unclear. At the single cell level, it is predicted that bacteria are less gravity-sensitive than larger species. The effects on their immediate environment, including the lack of cell settlement and slower mass transfer of nutrients due to lack of density driven convection, could help explain the trends. Ground-based spaceflight analogs, or simulated microgravity devices, are often employed to achieve different attributes of weightlessness to study effects on bacterial growth. Though these technologies could isolate gravity’s role in various biological processes, they cannot replicate all its effects and underlying mechanisms. Hence, interpretation of results could be misleading, even if similar to spaceflight. In this study two common simulated microgravity devices were investigated to determine whether they could simulate relevant microgravity conditions for bacterial growth. A bioreactor, the high aspect ratio vessel (HARV), was used with dyes of different density mounted on a random positioning machine (RP machine) or a rotating wall vessel (RWV). The RP machine displayed higher mixing rates than the RWV. The RWV was further tested at different rotations per minute (RPM). The range to minimize effects of density driven convection (low speeds) or centrifugal forces (high speeds) was between a range of 15-20 RPM. These results will help inform the selection of simulated microgravity device as well as interpretation of subsequent biofilm growth results.

Angie M. Diaz↗

NASA NTRSDate not supplied

Effects of Spaceflight Relevant Carbon Dioxide Levels on Pathogenesis Related Microbial Characteristics

Multiple stressors in the spaceflight environment, including microgravity and radiation, have the potential to alter microbial pathogenesis and virulence characteristics. Understanding how these and other factors may exacerbate the risk of infectious disease for astronauts is important to ensure the success of the mission. One understudied question in our assessment of infectious disease risk is the contribution carbon dioxide (CO2) at atmospheric concentrations higher than would be observed in terrestrial settings. Previous terrestrial studies have shown altered gene expression, selective bacterial inhibition, increased growth and diversity, and increased antibiotic resistance of bacterial communities individually and in biofilms when exposed to increased levels of CO2. While many of these findings are based on levels of CO2 higher than would be found during spaceflight exploration missions, the potential for subtle changes in CO2 levels to exacerbate infectious disease risks warrants an evaluation of pathogen responses to spaceflight CO2 conditions. To address this question, we are evaluating the effects of elevated atmospheric CO2 on microbial concentration, diversity, and pathogenesis-related characteristics using medically significant microorganisms cultured at CO2 levels representing terrestrial atmospheric concentration (≈0.04 %), ISS elevated concentration (0.4 %), and an unexpectedly elevated concentration (1 %) to represent a worst-case scenario. To determine if microbial responses to these CO2 concentrations may be exacerbated in response to the spaceflight environment, microorganisms are being cultured under static conditions and both spaceflight analogue and control conditions using the Rotating Wall Vessel (RWV) bioreactor (Synthecon, Houston, TX). For these studies, we will evaluate the response of Staphylococcus aureus, Streptococcus pneumoniae, and Enterobacter aerogenes, which are opportunistic bacterial pathogens that are of medical significance and have been or are likely to be found aboard human-inhabited spacecraft. Bacteria will be evaluated individually and together as a consortium. The following Specific Aims are be investigated: Aim 1. Characterize the growth, diversity, and pathogenesis-related stress responses of medically significant microorganisms grown under multiple CO2 concentrations under static growth in a liquid medium. Stress responses include acid stress, oxidative stress, and thermal stress. Aim 2. Characterize the growth, diversity, and pathogenesis-related stress responses of medically significant microorganisms grown under multiple CO2 concentrations in both spaceflight analogue and control conditions. Stress responses include acid stress, oxidative stress, and thermal stress.

S G Thornhill↗

NASA NTRSDate not supplied

Space Algae-2: Preflight Testing for A Long-Duration, Multi-Omics Analysis of Arthrospira Platensis

The cyanobacteria Arthrospira platensis NIES-39, commonly known as spirulina, could provide a fresh supply of nutrients for crew on long-duration spaceflight missions. Spirulina is a readily digestible food that is high in protein with all essential amino acids as well as significant levels of B vitamins, antioxidants, and anti-inflammatory metabolites. Spaceflight has multiple abiotic stressors such as increased ionizing radiation and microgravity, which causes a lack of convective mixing. These environmental conditions may impact productivity, nutritional composition, and in long-duration propagation, spaceflight stress may impact the genetic stability of spirulina cultures. We are developing an International Space Station experiment to continuously culture A. platensis for six months. Multi-omics profiling will be used to monitor for changes in the genome, transcriptome, proteome, and metabolome to determine if A. platensis is a suitable nutritional supplement on long-duration missions. During preflight testing we developed a protocol for inoculated liquid cultures to survive a 10-week storage period prior to photo-incubation. The bioreactor bag, temperature, and lighting conditions that support a 14-day growth cycle between passages were also determined. Media testing identified minimal salts supporting robust growth that can be stored in liquid or dry form. A simple filtration method was developed to dewater cultures and harvest biomass for frozen sample return. We optimized a cryopreservation method to enable return of live cells for isolation of individual A. platensis clones. The concept of operations for Space Algae-2 developed from these test results as well as progress on multi-omics analysis methods will be presented.

Algae↗

NASA NTRS2024

Impact of Simulated Microgravity on Mesenchymal Stem Cell Proliferation

Background Long term exposure to microgravity has adverse effects on human body such as bone loss. Due to the vital role of mesenchymal stem cell (MSC) in bone regeneration, studying MSC under microgravity conditions has been of interest during the past decade. However, high cost and limited access to space flight can dampen enthusiasm of researchers. Microgravity simulators provide a way to investigate the effects of microgravity in human physiology. Methods In this study, we compared MSC cultures under simulated microgravity using random positioning machine (RPM) and Gravite 3D simulators. Two culture durations, 1 week and 2 weeks, were selected for MSC cultures without media change. To evaluate impact of simulated microgravity on cell proliferation, cell counting was performed using trypan blue exclusion assay. Additionally, RNA samples and conditioned media were collected for further analysis. Results After 1 week and 2 weeks, no cell morphology change was observed from RPM and Gravite cultures compared to control. Notably, distinct lower cell confluence at the edge of the flasks on the RPM was observed, which could be caused by fluid sheer forces. MSCs cultured on both microgravity simulators showed decreased cell proliferation after 1-week culture. After 2-week culture, cells grown on the RPM showed no significant difference in cell proliferation compared with control while cells cultured on the Gravite bioreactor showed significantly enhanced MSC proliferation. Conclusion Type of microgravity simulator and duration of culture significantly affect MSC proliferation. Further investigation into how simulated microgravity influences MSC gene expression and cytokine secretion related to bone loss is currently on going in our lab.

Cuiping Zhang↗

NASA NTRSDate not supplied

Space Algae-2: Preflight Testing for A Long-Duration, Multi-Omics Analysis of Arthrospira Platensis

Algae↗

NASA NTRS2024

Space Algae-2 Ground and Lunar Analog Studies in Preparation for Long-Duration Propagation of Cyanobacteria in Spaceflight

There are numerous applications for microalgae in spaceflight missions and on Earth, such as oxygen production, carbon dioxide removal, nutrition, wastewater processing, and biofuel production. Space Algae-2 aims to test the genetic stability of Arthrospira platensis, commonly known as spirulina, during six-months of continuous culture on the International Space Station. Long-duration exposure to ionizing radiation and microgravity may impact growth, nutrient composition, and genetic stability. The high protein, vitamin, antioxidant content, and radiation resistance make spirulina a promising candidate for bioregenerative life support systems. A concept of operations was developed to grow and harvest algal biomass in space. Preflight testing experiments optimized conditions for an extended growth period in a gas permeable bioreactor bag. Preflight and post-harvest storage methods were developed in addition to a novel cryopreservation method. After sample return, multi-omics analysis will be conducted to determine the mutation rate, gene expression, and the protein and metabolite profile. The concept of operations for Space Algae-2 was tested during a lunar mission simulation within a semi-controlled environment. During a six-day lunar analog mission at the Hawai’i Space Exploration Analog and Simulation (HI-SEAS) A. platensis was successfully grown using flight-like hardware. The cyanobacteria were harvested and used to supplement bread as an example of spirulina biomass utilization. Overall, the data collected from Space Algae-2 will inform potential bioengineering of spirulina for space and terrestrial applications.

Algae↗

NASA NTRSDate not supplied

Space Algae-2 Ground and Lunar Analog Studies in Preparation for Long-Duration Propagation of Cyanobacteria in Spaceflight

There are numerous applications for microalgae in spaceflight missions and on Earth, such as, oxygen production, carbon dioxide removal, nutrition, wastewater processing, and biofuel production. Space Algae-2 aims to test the genetic stability of Arthrospira platensis, commonly known as spirulina, during six-months of continuous culture in spaceflight on the International Space Station. Long-duration exposure to ionizing radiation and microgravity may impact growth, nutrient composition, and genetic stability. The high protein, vitamin, antioxidant content, and radiation resistance make spirulina a promising candidate for bioregenerative life support systems during long-duration missions. A concept of operations was developed to grow and harvest algal biomass in space. Preflight testing experiments were conducted to optimize conditions for an extended growth period in a gas permeable bioreactor bag. Preflight and post-harvest storage methods were developed in addition to a novel cryopreservation method. After sample return, multi-omics analysis will be conducted to determine the mutation rate, gene expression, and protein and metabolite profile. The concept of operations for Space Algae-2 was tested at HI-SEAS (Hawai’i Space Exploration Analog and Simulation) during a six-day lunar analog mission (EMMIHS23, EuroMoonMars, International MoonBase Alliance, HI-SEAS, 2023). A. platensis was grown in the semi-controlled environment using flight-like hardware and solar powered LED lights. Then, the biomass was harvested and used to supplement bread as an example of A. platensis utilization. Overall, the data collected from Space Algae-2 will inform potential bioengineering of spirulina for space and terrestrial applications.

Algae↗

NASA NTRSDate not supplied

CO2-Based Manufacturing System for Recombinant Protein Production

Space biomanufacturing is a potential In Situ Resource Utilization (ISRU) strategy to provide critical consumables and products while minimizing the launched mass for long-duration, deep space missions. On Earth, the primary biological conversion of CO 2 to biomass is through photosynthesis, and sugars from photosynthetic organisms are used as feedstocks for microbial biomanufacturing. The efficiency of non-biological reduction of CO 2 to organic molecules, such as acetate or ethanol, has greatly increased in recent years. We are designing a biomanufacturing system to rely on electrochemical CO 2 conversion products for carbon substrates to support microbial growth and production of recombinant proteins. The preliminary design includes a gas-permeable membrane bioreactor with dry salts that are rehydrated and mixed with the carbon source to support growth of bacteria or yeast. The system architecture has a partially automated bioprocessing system to concentrate biomass and purify recombinant protein. This system is designed to operate semi-autonomously with minimal crew intervention. The specific use-case scenario is to produce a thermal stable carbonic anhydrase to increase the efficiency of a proposed liquid amine CO 2 removal subsystem of an environmental control and life-support system (ECLSS) on Mars.

Biomanufacturing↗