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Synthetic Biology and Microbial Fuel Cells: Towards Self-Sustaining Life Support Systems

NASA ARC and the J. Craig Venter Institute (JCVI) collaborated to investigate the development of advanced microbial fuels cells (MFCs) for biological wastewater treatment and electricity production (electrogenesis). Synthetic biology techniques and integrated hardware advances were investigated to increase system efficiency and robustness, with the intent of increasing power self-sufficiency and potential product formation from carbon dioxide. MFCs possess numerous advantages for space missions, including rapid processing, reduced biomass and effective removal of organics, nitrogen and phosphorus. Project efforts include developing space-based MFC concepts, integration analyses, increasing energy efficiency, and investigating novel bioelectrochemical system applications

bioelectrochemical systems↗

Interaction and Synergism of Microbial Fuel Cell Bacteria within Methanogenesis

Biological hydrogen production from waste biomass has both terrestrial and Martian advanced life support applications. On earth, biological hydrogen production is being explored as a greenhouse neutral form of clean and efficient energy. In a permanently enclosed space habitat, carbon loop closure is required to reduce mission costs. Plants are grown to revitalize oxygen supply and are consumed by habitat inhabitants. Unharvested portions must then be recycled for reuse in the habitat. Several biological degradation techniques exist, but one process, biophotolysis, can be used to produce hydrogen from inedible plant biomass. This process is two-stage, with one stage using dark fermentation to convert plant wastes into organic acids. The second stage, photofermentation, uses photoheterotrophic purple non-sulfur bacteria with the addition of light to turn the organic acids into hydrogen and carbon dioxide. Such a system can prove useful as a co-generation scheme, providing some of the energy needed to power a larger primary carbon recovery system, such as composting. Since butyrate is expected as one of the major inputs into photofermentation, a characterization study was conducted with the bacterium Rhodobacter sphaeroides SCJ, a novel photoheterotrophic non-sulfur purple bacteria, to examine hydrogen production performance at 10 mM-100 mM butyrate concentrations. As butyrate levels increased, hydrogen production increased up to 25 mM, and then decreased and ceased by 100 mM. Additionally, lag phase increased with butyrate concentration, possibly indicating some product inhibition. Maximal substrate conversion efficiency was 8.0%; maximal light efficiency was 0.89%; and maximal hydrogen production rate was 7.7 Umol/mg/cdw/hr (173 ul/mg cdw/hr). These values were either consistent or lower than expected from literature.

Klaus, David↗

Low Power Microrobotics Utilizing Biologically Inspired Energy Generation

The Low Power Microrobotics (LPM) project is a NASA NIAC-funded multi-disciplinary research effort involving the research and advancement of low power electronics, low power mobility and their integration with state of the art microbial fuel cell for power generation. The goal of this project was to advance the capabilities of each of these systems with a 10-year future target of a near one kilogram space exploration vehicle that contains each of these systems. In order to accomplish this task, each system was investigated independently in order to establish the power requirements/limitations while considering how each system affects the other systems. This was performed through numerous trade studies and experimental work, primarily focused around the power capacity of the microbial fuel cell base-lined for this project and how its strengths and limitations affected the design of the remaining onboard systems. Finally an end-to-end system test utilized a microbial fuel cell to charge a super capacitor through an energy harvesting circuit, which was discharged to activate a robotic locomotion system.

Microrobotics↗

Cosmo Cassette: A Microfluidic Microgravity Microbial System For Synthetic Biology Unit Tests and Satellite Missions

Although methods in the design-build-test life cycle of the synthetic biology field have grown rapidly, the expansion has been non-uniform. The design and build stages in development have seen innovations in the form of biological CAD and more efficient means for building DNA, RNA, and other biological constructs. The testing phase of the cycle remains in need of innovation. Presented will be both a theoretical abstraction of biological measurement and a practical demonstration of a microfluidics-based platform for characterizing synthetic biological phenomena. Such a platform demonstrates a design of additive manufacturing (3D printing) for construction of a microbial fuel cell (MFC) to be used in experiments carried out in space. First, the biocompatibility of the polypropylene chassis will be demonstrated. The novel MFCs will be cheaper, and faster to make and iterate through designs. The novel design will contain a manifold switchingdistribution system and an integrated in-chip set of reagent reservoirs fabricated via 3D printing. The automated nature of the 3D printing yields itself to higher resolution switching valves and leads to smaller sized payloads, lower cost, reduced power and a standardized platform for synthetic biology unit tests on Earth and in space. It will be demonstrated that the application of unit testing in synthetic biology will lead to the automatic construction and validation of desired constructs. Unit testing methodologies offer benefits of preemptive problem identification, change of facility, simplicity of integration, ease of documentation, and separation of interface from implementation, and automated design.

Bioelectrochemical System↗

Recent experiences with iodine water disinfection in Shuttle

Microbial proliferation in the STS potable water system is prevented by maintaining a 2-5 ppm iodine residual. The iodine is added to fuel cell water by an iodinated ion exchange resin in the Microbial Check Valve (MCV). Crew comments indicated excessive iodine in the potable water. To better define the problem, a method of in-flight iodine analysis was developed. Inflight analysis during STS-30 and STS-28 indicated iodine residuals were generally in the 9-13 ppm range. It was determined that the high iodine residual was caused by MCV influent temperatures in excess of 120 F. This is well above the MCV operating range of 65-90 F. The solution to this problem was to develop a resin suitable for the higher temperatures. Since 8 months were required to formulate a MCV resin suitable for the higher temperatures, a temporary solution was necessary. Two additional MCV's were installed on the chilled and ambient water lines leading into the galley to remove the excess iodine. These reduced the iodine residual to 3-4 ppm during STS-33, STS-34, STS-36 and STS-32. A high-temperature resin was formulated and initially flown on STS-31.

Gibbons, Randall E.↗

Microbial Surveillance of Potable Water Sources of the International Space Station

To mitigate risk to the crew, the microbial surveillance of the quality of potable water sources of the International Space Station (ISS) has been ongoing since before the arrival of the first permanent crew. These water sources have included stored ground-supplied water, water produced by the shuttle fuel cells during flight, and ISS humidity condensate that is reclaimed and processed. Monitoring was accomplished using a self-contained filter designed to allow bacterial growth and enumeration during flight. Upon return to earth, microbial isolates were identified using 16S ribosomal gene sequencing. While the predominant isolates were common Gramnegative bacteria including Ralstonia eutropha, Methylobacterium fujisawaense, and Spingomonas paucimobilis, opportunistic pathogens such as Stenotrophomonas maltophilia and Pseudomonas aeruginosa were also isolated. Results of in-flight enumeration have indicated a fluctuation of bacterial counts above system design specifications. Additional in-flight monitoring capability for the specific detection of coliforms was added in 2004; no coliforms have been detected from any potable water source. Neither the bacterial concentrations nor the identification of the isolates recovered from these samples has suggested a threat to crew health.

Bruce, Rebekah J.↗

Constructing COMSOL Models of a Bacteriological Fuel Cell

We show very initial work on a specific bioelectrochemical system (BES), a bacteriologically driven 'fuel cell' (BFS), that is intended to process waste products, such as CO2 and brine. (1) Processing is the priority, not power generation (2) Really a Microbial Electrolysis Cell (MEC)

Coker, Robert↗

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.↗

Apollo experience report: Potable water system

A description of the design and function of the Apollo potable water system is presented. The command module potable water is supplied as a byproduct of the fuel cells. The cells, located in the service module, function primarily to supply electrical energy to the spacecraft. The source of the lunar module potable water is three tanks, which are filled before lift-off. The technique of supplying the water in each of these cases and the problems associated with materials compatibility are described. The chemical and microbiological quality of the water is reviewed, as are efforts to maintain the water in a microbially safe condition for drinking and food mixing.

Sauer, R. L.↗

Analysis of MIR Condensate and Potable Water

Approximately fifty percent of the potable water supplied to the Russian cosmonauts, American astronauts, and other occupants of the current Russian Mir Space Station is produced by the direct recycle of water from humidity condensate. The remainder comes from ground supplied potable water that is delivered on a Progress resupply spacecraft, or processed fuel cell water transferred from the Shuttle. Reclamation of water for potable and hygiene purposes is considered essential for extended duration missions in order to avoid massive costs associated with resupplying water from the ground. The Joint U.S/Russian Phase 1 program provided the U.S. the first opportunity to evaluate the performance of water reclamation hardware in microgravity. During the Phase I program, the U.S. collected recycled water, stored water, and humidity condensate samples for chemical and microbial evaluation. This experiment was conducted to determine the potability of the water supplied on Mir, to assess the reliability of the water reclamation and distribution systems, and to aid in developing water quality monitoring standards for International Space Station.

Pierre, L. M.↗

NASA Tech Briefs, May 2013

Topics include: Test Waveform Applications for JPL STRS Operating Environment; Pneumatic Proboscis Heat-Flow Probe; Method to Measure Total Noise Temperature of a Wireless Receiver During Operation; Cursor Control Device Test Battery; Functional Near-Infrared Spectroscopy Signals Measure Neuronal Activity in the Cortex; ESD Test Apparatus for Soldering Irons; FPGA-Based X-Ray Detection and Measurement for an X-Ray Polarimeter; Sequential Probability Ratio Test for Spacecraft Collision Avoidance Maneuver Decisions; Silicon/Carbon Nanotube Photocathode for Splitting Water; Advanced Materials and Fabrication Techniques for the Orion Attitude Control Motor; Flight Hardware Packaging Design for Stringent EMC Radiated Emission Requirements; RF Reference Switch for Spaceflight Radiometer Calibration; An Offload NIC for NASA, NLR, and Grid Computing; Multi-Scale CNT-Based Reinforcing Polymer Matrix Composites for Lightweight Structures; Ceramic Adhesive and Methods for On-Orbit Repair of Re-Entry Vehicles; Self-Healing Nanocomposites for Reusable Composite Cryotanks; Pt-Ni and Pt-Co Catalyst Synthesis Route for Fuel Cell Applications; Aerogel-Based Multilayer Insulation with Micrometeoroid Protection; Manufacturing of Nanocomposite Carbon Fibers and Composite Cylinders; Optimized Radiator Geometries for Hot Lunar Thermal Environments; A Mission Concept: Re-Entry Hopper-Aero-Space-Craft System on-Mars (REARM-Mars); New Class of Flow Batteries for Terrestrial and Aerospace Energy Storage Applications; Reliability of CCGA 1152 and CCGA 1272 Interconnect Packages for Extreme Thermal Environments; Using a Blender to Assess the Microbial Density of Encapsulated Organisms; Mixed Integer Programming and Heuristic Scheduling for Space Communication; Video Altimeter and Obstruction Detector for an Aircraft; Control Software for Piezo Stepping Actuators; Galactic Cosmic Ray Event-Based Risk Model (GERM) Code; Sasquatch Footprint Tool; and Multi-User Space Link Extension (SLE) System.

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Advanced Microbial Check Valve development

The Microbial Check Valve (MCV) is a flight qualified assembly that provides bacteriologically safe drinking water for the Space Shuttle. The 1-lb unit is basically a canister packed with an iodinated ion-exchange resin. The device is used to destroy organisms in a water stream as the water passes through it. It is equally effective for fluid flow in either direction and its primary method of disinfection is killing rather than filtering. The MCV was developed to disinfect the fuel cell water and to prevent back contamination of stored potable water on the Space Shuttle. This paper reports its potential for space applications beyond the basic Shuttle mission. Data are presented that indicate the MCV is suitable for use in advanced systems that NASA has under development for the reclamation of humidity condensate, wash water and human urine.

Colombo, G. V.↗

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.↗

Constraining Venus Cloud Habitability: Earth's Aerobiosphere as an Analogue Environment

The potential habitability of Venus's cloud and haze layers has been debated for decades, with interest fueled by observations including disequilibria in atmospheric chemistry, strongly UV-absorbing particles, and (most recently) a controversial phosphine detection. Venus cloud temperature and pressure are clement by Earth standards; however, the calculated high acid activity and low water activity may exceed the limits of Earth biochemistry, based on observed life in extreme environments. A significant challenge in understanding Venus's habitability is the lack of an appropriate terrestrial analogue environment. Life has not been found in hydrothermal systems approaching Venus aerosol acidities, but these systems also contain high levels of other harsh solutes. Earth's stratospheric sulfate layer is a partial match in terms of acidity, desiccation, size, and isolation from the surface, but the few samples returned from these altitudes have yielded only sparse, inactive cells. Earth's tropospheric cloud droplets have an active microbial presence, but are larger and far more water-rich than Venus aerosols; tropospheric times aloft are also similar to microbial generation times, making Earth's aerobiosphere dependent on continuous surface replenishment. Direct detection of potential biosignatures from a single transect is challenging; Earth's cloud microbiota yields 102 - 108 viable cells per mL, equivalent to one per 103 - 109 particles, with highly dynamic and heterogeneous distribution. The most important in situ measurement for Venus cloud habitability is detailed aerosol composition: confirmation of acid and water activity, presence of potentially bioavailable nutrients and energy, and potential presence of organic matter. Modeling Venus cloud aerosol residence time in comparison to periodic influxes of water and photochemical energy, and thus the ability of an ecosystem to maintain itself over time, would also address a significant habitability constraint particular to aerobiospheres.

Venus↗

PowerCell aboard the EuCROPIS satellite - Results from the first synthetic biology experiment in space

Synthetic biology is the design and construction of new biological parts and systems not found in nature, a technology that we believe will be a game-changing technology for space exploration. The PowerCell concept is an ecology based on photosynthetic microbes taking advantage of in situ materials and energy to generate, on-demand, useful products (food, fuel, cloths, drugs, etc.) that satisfy specific needs of long-term human presence away from Earth. This first PowerCell experiment is testing this concept as well as several key mechanisms of engineering biology in a 48-well plate microfluidics system. NASA's PowerCell experiment was loaded in the fluidics system at NASA Ames in May 2016, and subsequently integrated on the DLR's compact satellite Eu:CROPIS (Euglena Combined Regenerative Organic food Production In Space), as a secondary payload. Launched into Earth’s orbit from the Vandenberg Air Force Base, California, USA, in December 3, 2019, the Eu:CROPIS platform has provided an artificial gravity through rotational acceleration, simulating gravity ranges compatible to those found on Mars and on the Moon, as well as microgravity in the absence of acceleration. Each set of biological experiments performed at Earth's orbit was compared to ground controls. Over 400 days' worth of mission data showed that all hardware and software worked as planned, and microbial growth was observed in all four gravity regimes, including growth on cyanobacterial cell extract in three out of four gravity regimes. The genetic transformation experiment was successful in microgravity and provided valuable long-term reagent stability data, including antibiotic efficacy. The demonstrated ability of hardy bacterial spores to recover from a long-term stasis period will be fundamental for deep human space missions. Lessons learned from hardware design to experimental operations are leading to improved designs and capabilities for future missions.

Ivan Glaucio Paulino Lima↗

NASA Tech Briefs, September 2012

Topics covered include: Beat-to-Beat Blood Pressure Monitor; Measurement Techniques for Clock Jitter; Lightweight, Miniature Inertial Measurement System; Optical Density Analysis of X-Rays Utilizing Calibration Tooling to Estimate Thickness of Parts; Fuel Cell/Electrochemical Cell Voltage Monitor; Anomaly Detection Techniques with Real Test Data from a Spinning Turbine Engine-Like Rotor; Measuring Air Leaks into the Vacuum Space of Large Liquid Hydrogen Tanks; Antenna Calibration and Measurement Equipment; Glass Solder Approach for Robust, Low-Loss, Fiber-to-Waveguide Coupling; Lightweight Metal Matrix Composite Segmented for Manufacturing High-Precision Mirrors; Plasma Treatment to Remove Carbon from Indium UV Filters; Telerobotics Workstation (TRWS) for Deep Space Habitats; Single-Pole Double-Throw MMIC Switches for a Microwave Radiometer; On Shaft Data Acquisition System (OSDAS); ASIC Readout Circuit Architecture for Large Geiger Photodiode Arrays; Flexible Architecture for FPGAs in Embedded Systems; Polyurea-Based Aerogel Monoliths and Composites; Resin-Impregnated Carbon Ablator: A New Ablative Material for Hyperbolic Entry Speeds; Self-Cleaning Particulate Prefilter Media; Modular, Rapid Propellant Loading System/Cryogenic Testbed; Compact, Low-Force, Low-Noise Linear Actuator; Loop Heat Pipe with Thermal Control Valve as a Variable Thermal Link; Process for Measuring Over-Center Distances; Hands-Free Transcranial Color Doppler Probe; Improving Balance Function Using Low Levels of Electrical Stimulation of the Balance Organs; Developing Physiologic Models for Emergency Medical Procedures Under Microgravity; PMA-Linked Fluorescence for Rapid Detection of Viable Bacterial Endospores; Portable Intravenous Fluid Production Device for Ground Use; Adaptation of a Filter Assembly to Assess Microbial Bioburden of Pressurant Within a Propulsion System; Multiplexed Force and Deflection Sensing Shell Membranes for Robotic Manipulators; Whispering Gallery Mode Optomechanical Resonator; Vision-Aided Autonomous Landing and Ingress of Micro Aerial Vehicles; Self-Sealing Wet Chemistry Cell for Field Analysis; General MACOS Interface for Modeling and Analysis for Controlled Optical Systems; Mars Technology Rover with Arm-Mounted Percussive Coring Tool, Microimager, and Sample-Handling Encapsulation Containerization Subsystem; Fault-Tolerant, Real-Time, Multi-Core Computer System; Water Detection Based on Object Reflections; SATPLOT for Analysis of SECCHI Heliospheric Imager Data; Plug-in Plan Tool v3.0.3.1; Frequency Correction for MIRO Chirp Transformation Spectroscopy Spectrum; Nonlinear Estimation Approach to Real-Time Georegistration from Aerial Images; Optimal Force Control of Vibro-Impact Systems for Autonomous Drilling Applications; Low-Cost Telemetry System for Small/Micro Satellites; Operator Interface and Control Software for the Reconfigurable Surface System Tri-ATHLETE; and Algorithms for Determining Physical Responses of Structures Under Load.

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NASA Tech Briefs, January 2013

Topics include: Single-Photon-Sensitive HgCdTe Avalanche Photodiode Detector; Surface-Enhanced Raman Scattering Using Silica Whispering-Gallery Mode Resonators; 3D Hail Size Distribution Interpolation/Extrapolation Algorithm; Color-Changing Sensors for Detecting the Presence of Hypergolic Fuels; Artificial Intelligence Software for Assessing Postural Stability; Transformers: Shape-Changing Space Systems Built with Robotic Textiles; Fibrillar Adhesive for Climbing Robots; Using Pre-Melted Phase Change Material to Keep Payloads in Space Warm for Hours without Power; Development of a Centrifugal Technique for the Microbial Bioburden Analysis of Freon (CFC-11); Microwave Sinterator Freeform Additive Construction System (MS-FACS); DSP/FPGA Design for a High-Speed Programmable S-Band Space Transceiver; On-Chip Power-Combining for High-Power Schottky Diode-Based Frequency Multipliers; FPGA Vision Data Architecture; Memory Circuit Fault Simulator; Ultra-Compact Transputer-Based Controller for High-Level, Multi-Axis Coordination; Regolith Advanced Surface Systems Operations Robot Excavator; Magnetically Actuated Seal; Hybrid Electrostatic/Flextensional Mirror for Lightweight, Large-Aperture, and Cryogenic Space Telescopes; System for Contributing and Discovering Derived Mission and Science Data; Remote Viewer for Maritime Robotics Software; Stackfile Database; Reachability Maps for In Situ Operations; JPL Space Telecommunications Radio System Operating Environment; RFI-SIM: RFI Simulation Package; ION Configuration Editor; Dtest Testing Software; IMPaCT - Integration of Missions, Programs, and Core Technologies; Integrated Systems Health Management (ISHM) Toolkit; Wind-Driven Wireless Networked System of Mobile Sensors for Mars Exploration; In Situ Solid Particle Generator; Analysis of the Effects of Streamwise Lift Distribution on Sonic Boom Signature; Rad-Tolerant, Thermally Stable, High-Speed Fiber-Optic Network for Harsh Environments; Towed Subsurface Optical Communications Buoy; High-Collection-Efficiency Fluorescence Detection Cell; Ultra-Compact, Superconducting Spectrometer-on-a-Chip at Submillimeter Wavelengths; UV Resonant Raman Spectrometer with Multi-Line Laser Excitation; Medicine Delivery Device with Integrated Sterilization and Detection; Ionospheric Simulation System for Satellite Observations and Global Assimilative Model Experiments - ISOGAME; Airborne Tomographic Swath Ice Sounding Processing System; flexplan: Mission Planning System for the Lunar Reconnaissance Orbiter; Estimating Torque Imparted on Spacecraft Using Telemetry; PowderSim: Lagrangian Discrete and Mesh-Free Continuum Simulation Code for Cohesive Soils; Multiple-Frame Detection of Subpixel Targets in Thermal Image Sequences; Metric Learning to Enhance Hyperspectral Image Segmentation; Basic Operational Robotics Instructional System; Sheet Membrane Spacesuit Water Membrane Evaporator; Advanced Materials and Manufacturing for Low-Cost, High-Performance Liquid Rocket Combustion Chambers; Motor Qualification for Long-Duration Mars Missions.

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