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NASA NTRS1977

Microbiological profiles of the Viking spacecraft

Planetary quarantine requirements associated with the launch of two Viking spacecraft necessitated microbiological assessment during assembly and testing at Cape Canaveral and the Kennedy Space Center. Samples were collected from the Viking Lander Capsules (VLC), Orbiters (VO), and Shrouds at predetermined intervals during assembly and testing. Levels of bacterial spores per square meter on the VLC-1 and VLC-2 were 1.6 x 10 squared and 9.7, respectively, prior to dry-heat sterilization. The ranges of aerobic mesophilic microorganisms detected on the VO-1 and VO-2 were 4.2 x 10 squared to 4.3 x 10 cubed and 2.3 x 10 squared to 8.9 x 10 cubed/sq m, respectively. Approximately 1300 colonies were picked from culture plates, identified, lyophilized, and stored for future reference. About 75% of all isolates were microorganisms considered indigenous to humans; the remaining isolates were associated with soil and dust. The percentage of microorganisms of human origin was consistent with results obtained with previous automated spacecraft but slightly lower than those observed for manned (Apollo) spacecraft.

Puleo, J. R.↗
NASA NTRS1972

Microbiological sampling of returned Surveyor 3 electrical cabling

A piece of electrical wiring bundle running from the television camera to another part of the spacecraft was selected for microbiological examination. Sampling methods are discussed. The results presented show that no viable microorganisms were recovered from the part of the Surveyor 3 cable which was tested. Factors that could have contributed to the sterility of the cable are thermal vacuum testing, natural dieoff, change in pressure during launch, and lunar vacuum and temperature.

Knittel, M. D.↗
NASA NTRS2006

NASA Tech Briefs, July 2006

Topics covered include: Airport Remote Tower Sensor Systems; Implantable Wireless MEMS Sensors for Medical Uses; Embedded Sensors for Measuring Surface Regression; Coordinating an Autonomous Earth-Observing Sensorweb; Range-Measuring Video Sensors; Stability Enhancement of Polymeric Sensing Films Using Fillers; Sensors for Using Times of Flight to Measure Flow Velocities; Receiver Would Control Phasing of a Phased-Array Antenna; Modern Design of Resonant Edge-Slot Array Antennas; Carbon-Nanotube Schottky Diodes; Simplified Optics and Controls for Laser Communications; Coherent Detection of High-Rate Optical PPM Signals; Multichannel Phase and Power Detector; Using Satellite Data in Weather Forecasting: I; Using Dissimilarity Metrics to Identify Interesting Designs; X-Windows PVT Widget Class; Shuttle Data Center File-Processing Tool in Java; Statistical Evaluation of Utilization of the ISS; Nanotube Dispersions Made With Charged Surfactant; Aerogels for Thermal Insulation of Thermoelectric Devices; Low-Density, Creep-Resistant Single-Crystal Superalloys; Excitations for Rapidly Estimating Flight-Control Parameters; Estimation of Stability and Control Derivatives of an F-15; Tool for Coupling a Torque Wrench to a Round Cable Connector; Ultrasonically Actuated Tools for Abrading Rock Surfaces; Active Struts With Variable Spring Stiffness and Damping; Multiaxis, Lightweight, Computer-Controlled Exercise System; Dehydrating and Sterilizing Wastes Using Supercritical CO2; Alpha-Voltaic Sources Using Liquid Ga as Conversion Medium; Ice-Borehole Probe; Alpha-Voltaic Sources Using Diamond as Conversion Medium; White-Light Whispering-Gallery-Mode Optical Resonators; Controlling Attitude of a Solar-Sail Spacecraft Using Vanes; and Wire-Mesh-Based Sorber for Removing Contaminants from Air.

Source record↗
NASA NTRS2012

The Mars Sample Return Lab(s) - Lessons from the Past and Implications for the Future

It has been widely understood for many years that an essential component of a Mars Sample Return mission is a Sample Receiving Facility (SRF). The purpose of such a facility would be to take delivery of the flight hardware that lands on Earth, open the spacecraft and extract the sample container and samples, and conduct an agreed upon test protocol, while ensuring strict containment and contamination control of the samples while in the SRF. Any samples that are found to be non-hazardous (or are rendered non-hazardous by sterilization) would then be transferred to long-term curation. Although the general concept of an SRF is relatively straightforward, there has been considerable discussion about implementation planning.

Allen, Carlton↗
NASA NTRS2013

Characterization of Heat Melt Compactor (HMC) Product Water

The Heat Melt Compactor (HMC) is designed to sterilize and process wastes produced during space missions. Benefits of the HMC include reduction of biohazards to the crew, reduction in volume of wastes that would otherwise require storage, production of radiation shielding tiles, and recovery of water and other resources. Water reuse is critical onboard spacecrafts; it reduces the need for resupply missions and saves valuable storage space. The main sources of water in HMC batches are food, beverages, shampoo, disinfecting wipes, toothpaste, and diapers. Water reclaimed by the HMC was analyzed for concentrations of Na+, NH4+, K+, Mg2+, Ca2+, Cl-­‐, NO2-­‐, Br-­‐, NO3-­‐, PO43-­‐, SO42-­‐, total organic carbon (TOC), total inorganic carbon (TIC), % total solids, and pH. The data are discussed in relation to the current water input characteristics established for the International Space Station Water Processor Assembly system. Batches with higher than average amounts of food produced HMC product water with higher sulfate content, and batches with higher proportions of disinfectant wipes and food yielded HMC product water with higher ammonium concentration. We also compared theoretical chemical composition of HMC product water based on food labels and literature values to experimental results.

Heat Melt Compactor↗
NASA NTRS2012

Extreme Ionizing-Radiation-Resistant Bacterium

There is a growing concern that desiccation and extreme radiation-resistant, non-spore-forming microorganisms associated with spacecraft surfaces can withstand space environmental conditions and subsequent proliferation on another solar body. Such forward contamination would jeopardize future life detection or sample return technologies. The prime focus of NASA s planetary protection efforts is the development of strategies for inactivating resistance-bearing microorganisms. Eradification techniques can be designed to target resistance-conferring microbial populations by first identifying and understanding their physiologic and biochemical capabilities that confers its elevated tolerance (as is being studied in Deinococcus phoenicis, as a result of this description). Furthermore, hospitals, food, and government agencies frequently use biological indicators to ensure the efficacy of a wide range of radiation- based sterilization processes. Due to their resistance to a variety of perturbations, the non-spore forming D. phoenicis may be a more appropriate biological indicator than those currently in use. The high flux of cosmic rays during space travel and onto the unshielded surface of Mars poses a significant hazard to the survival of microbial life. Thus, radiation-resistant microorganisms are of particular concern that can survive extreme radiation, desiccation, and low temperatures experienced during space travel. Spore-forming bacteria, a common inhabitant of spacecraft assembly facilities, are known to tolerate these extreme conditions. Since the Viking era, spores have been utilized to assess the degree and level of microbiological contamination on spacecraft and their associated spacecraft assembly facilities. Members of the non-spore-forming bacterial community such as Deinococcus radiodurans can survive acute exposures to ionizing radiation (5 kGy), ultraviolet light (1 kJ/sq m), and desiccation (years). These resistive phenotypes of Deinococcus enhance the potential for transfer, and subsequent proliferation, on another solar body such as Mars and Europa. These organisms are more likely to escape planetary protection assays, which only take into account presence of spores. Hence, presences of extreme radiation-resistant Deinococcus in the cleanroom facility where spacecraft are assembled pose a serious risk for integrity of life-detection missions. The microorganism described herein was isolated from the surfaces of the cleanroom facility in which the Phoenix Lander was assembled. The isolated bacterial strain was subjected to a comprehensive polyphasic analysis to characterize its taxonomic position. This bacterium exhibits very low 16SrRNA similarity with any other environmental isolate reported to date. Both phenotypic and phylogenetic analyses clearly indicate that this isolate belongs to the genus Deinococcus and represents a novel species. The name Deinococcus phoenicis was proposed after the Phoenix spacecraft, which was undergoing assembly, testing, and launch operations in the spacecraft assembly facility at the time of isolation. D. phoenicis cells exhibited higher resistance to ionizing radiation (cobalt-60; 14 kGy) than the cells of the D. radiodurans (5 kGy). Thus, it is in the best interest of NASA to thoroughly characterize this organism, which will further assess in determining the potential for forward contamination. Upon the completion of genetic and physiological characteristics of D. phoenicis, it will be added to a planetary protection database to be able to further model and predict the probability of forward contamination.

Vaishampayan, Parag A.↗
NASA NTRS2013

Extreme Ionizing-Radiation-Resistant Bacterium

There is a growing concern that desiccation and extreme radiation-resistant, non-spore-forming microorganisms associated with spacecraft surfaces can withstand space environmental conditions and subsequent proliferation on another solar body. Such forward contamination would jeopardize future life detection or sample return technologies. The prime focus of NASA s planetary protection efforts is the development of strategies for inactivating resistance-bearing micro-organisms. Eradi cation techniques can be designed to target resistance-conferring microbial populations by first identifying and understanding their physiologic and biochemical capabilities that confers its elevated tolerance (as is being studied in Deinococcus phoenicis, as a result of this description). Furthermore, hospitals, food, and government agencies frequently use biological indicators to ensure the efficacy of a wide range of radiation-based sterilization processes. Due to their resistance to a variety of perturbations, the nonspore forming D. phoenicis may be a more appropriate biological indicator than those currently in use. The high flux of cosmic rays during space travel and onto the unshielded surface of Mars poses a significant hazard to the survival of microbial life. Thus, radiation-resistant microorganisms are of particular concern that can survive extreme radiation, desiccation, and low temperatures experienced during space travel. Spore-forming bacteria, a common inhabitant of spacecraft assembly facilities, are known to tolerate these extreme conditions. Since the Viking era, spores have been utilized to assess the degree and level of microbiological contamination on spacecraft and their associated spacecraft assembly facilities. Members of the non-sporeforming bacterial community such as Deinococcus radiodurans can survive acute exposures to ionizing radiation (5 kGy), ultraviolet light (1 kJ/m2), and desiccation (years). These resistive phenotypes of Deinococcus enhance the potential for transfer, and subsequent proliferation, on another solar body such as Mars and Europa. These organisms are more likely to escape planetary protection assays, which only take into account presence of spores. Hence, presences of extreme radiation-resistant Deinococcus in the cleanroom facility where spacecraft are assembled pose a serious risk for integrity of life-detection missions. The microorganism described herein was isolated from the surfaces of the cleanroom facility in which the Phoenix Lander was assembled. The isolated bacterial strain was subjected to a comprehensive polyphasic analysis to characterize its taxonomic position. This bacterium exhibits very low 16SrRNA similarity with any other environmental isolate reported to date. Both phenotypic and phylogenetic analyses clearly indicate that this isolate belongs to the genus Deinococcus and represents a novel species. The name Deinococcus phoenicis was proposed after the Phoenix spacecraft, which was undergoing assembly, testing, and launch operations in the spacecraft assembly facility at the time of isolation. D. phoenicis cells exhibited higher resistance to ionizing radiation (cobalt-60; 14 kGy) than the cells of the D. radiodurans (5 kGy). Thus, it is in the best interest of NASA to thoroughly characterize this organism, which will further assess in determining the potential for forward contamination. Upon the completion of genetic and physiological characteristics of D. phoenicis, it will be added to a planetary protection database to be able to further model and predict the probability of forward contamination.

Vaishampayan, Parag A.↗
NASA NTRS2009

NASA Tech Briefs, November 2009

Topics covered include: Cryogenic Chamber for Servo-Hydraulic Materials Testing; Apparatus Measures Thermal Conductance Through a Thin Sample from Cryogenic to Room Temperature; Rover Attitude and Pointing System Simulation Testbed; Desktop Application Program to Simulate Cargo-Air-Drop Tests; Multimodal Friction Ignition Tester; Small-Bolt Torque-Tension Tester; Integrated Spacesuit Audio System Enhances Speech Quality and Reduces Noise; Hardware Implementation of a Bilateral Subtraction Filter; Simple Optoelectronic Feedback in Microwave Oscillators; Small X-Band Oscillator Antennas; Free-Space Optical Interconnect Employing VCSEL Diodes; Discrete Fourier Transform Analysis in a Complex Vector Space; Miniature Scroll Pumps Fabricated by LIGA; Self-Assembling, Flexible, Pre-Ceramic Composite Preforms; Flight-speed Integral Image Analysis Toolkit; Work Coordination Engine; Multi-Mission Automated Task Invocation Subsystem; Autonomously Calibrating a Quadrupole Mass Spectrometer; Determining Spacecraft Reaction Wheel Friction Parameters; Composite Silica Aerogels Opacified with Titania; Multiplexed Colorimetric Solid-Phase Extraction; Detecting Airborne Mercury by Use of Polymer/Carbon Films; Lattice-Matched Semiconductor Layers on Single Crystalline Sapphire Substrate; Pressure-Energized Seal Rings to Better Withstand Flows; Rollerjaw Rock Crusher; Microwave Sterilization and Depyrogenation System; Quantifying Therapeutic and Diagnostic Efficacy in 2D Microvascular Images; NiF2/NaF:CaF2/Ca Solid-State High-Temperature Battery Cells; Critical Coupling Between Optical Fibers and WGM Resonators; Microwave Temperature Profiler Mounted in a Standard Airborne Research Canister; Alternative Determination of Density of the Titan Atmosphere; Solar Rejection Filter for Large Telescopes; Automated CFD for Generation of Airfoil Performance Tables; Progressive Classification Using Support Vector Machines; Active Learning with Irrelevant Examples; A Data Matrix Method for Improving the Quantification of Element Percentages of SEM/EDX Analysis; Deployable Shroud for the International X-Ray Observatory; Improved Model of a Mercury Ring Damper; Optoelectronic pH Meter: Further Details; X-38 Advanced Sublimator; and Solar Simulator Represents the Mars Surface Solar Environment.

Source record↗
NASA NTRSDate not supplied

Gas Effluent Analysis of the Heat Melt Compactor

The Heat Melt Compactor (HMC) reduces volume, heat sterilizes, stabilizes, and manages gas and water effluent of the International Space Station (ISS) trash. Processing the trash at high temperatures produces volatile gas compounds that need to be treated before venting to cabin and/or the Vacuum Exhaust System (VES) in the ISS. The release of gases may not meet the Spacecraft Maximum Allowable Concentrations (SMAC) requirement, if vented directly into cabin, and/or gases are incompatible for venting to the VES. In order to assess the HMC gas release and venting parameters, effluent gas analysis is conducted to determine contaminant load. This paper will analyze different collection and analytical methods of the effluent gas, and report corresponding data, such that these results can be used for process design and effluent management.

Heat Melt Compactor↗
NASA NTRS1976

Labeled Release - An experiment in radiorespirometry

The Labeled Release extraterrestrial life detection experiment onboard the Viking spacecraft is described as it will be implemented on the surface of Mars in 1976. This experiment is designed to detect heterotrophic life by supplying a dilute solution of radioactive organic substrates to a sample of Martian soil and monitoring for evolution of radioactive gas. A significantly attenuated response by a heat-sterilized control sample of the same soil would confirm a positive metabolic response. Experimental assumptions as well as criteria for the selection of organic substrates are presented. The Labeled Release nutrient has been widely tested, is versatile in eliciting terrestrial metabolic responses, and is stable to heat sterilization and to the long-term storage required before its use on Mars. A testing program has been conducted with flight-like instruments to acquire science data relevant to the interpretation of the Mars experiment. Factors involved in the delineation of a positive result are presented and the significance of the possible results discussed.

Levin, G. V.↗
NASA NTRS1999

Growing crops for space explorers on the moon, Mars, or in space

An option in the long-duration exploration of space, whether on the Moon or Mars or in a spacecraft on its way to Mars or the asteroids, is to utilize a bioregenerative life-support system in addition to the physicochemical systems that will always be necessary. Green plants can use the energy of light to remove carbon dioxide from the atmosphere and add oxygen to it while at the same time synthesizing food for the space travelers. The water that crop plants transpire can be condensed in pure form, contributing to the water purification system. An added bonus is that green plants provide a familiar environment for humans far from their home planet. The down side is that such a bioregenerative life-support system--called a controlled environment life-support system (CELSS) in this paper--must be highly complex and relatively massive to maintain a proper composition of the atmosphere while also providing food. Thus, launch costs will be high. Except for resupply and removal of nonrecycleable substances, such a system is nearly closed with respect to matter but open with respect to energy. Although a CELSS facility is small compared to the Earth's biosphere, it must be large enough to feed humans and provide a suitable atmosphere for them. A functioning CELSS can only be created with the help of today's advanced technology, especially computerized controls. Needed are energy for light, possibly from a nuclear power plant, and equipment to provide a suitable environment for plant growth, including a way to supply plants with the necessary mineral nutrients. All this constitutes the biomass production unit. There must also be food preparation facilities and a means to recycle or dispose of waste materials and there must be control equipment to keep the facility running. Humans are part of the system as well as plants and possibly animals. Human brain power will often be needed to keep the system functional in spite of the best computer-driven controls. The particulars of a CELSS facility depend strongly on where it is to be located. The presence of gravity on the Moon and Mars simplifies the design for a facility on those bodies, but a spacecraft in microgravity is a much more challenging environment. One problem is that plants, which are very sensitive to gravity, might not grow and produce food in the virtual absence of gravity. However, the experience with growing super-dwarf wheat in the Russian space station Mir, while not entirely successful because of the sterile wheat heads, was highly encouraging. The plants grew well for 123 days, producing more biomass than had been produced in space before. This was due to the high photon flux available to the plants and the careful control of substrate moisture. The sterile heads were probably due to the failure to remove the gaseous plant hormone, ethylene, from the Mir atmosphere. Since ethylene can easily be removed, it should be possible to grow wheat and other crops in microgravity with the production of viable seeds. On the ground Biosphere-2 taught us several lessons about the design and construction of a CELSS facility, but Bios-3 came much closer to achieving the goals of such a facility. Although stability was never completely reached, Bios-3 was much more stable than Biosphere-2 apparently because every effort was made to keep the system simple and to use the best technology available to maintain control. Wastes were not recycled in Bios-3 except for urine, and inedible plant materials were incinerated to restore CO2 to the atmosphere. Since much meat (about 20% of calories) was imported, closure in the Bios-3 experiments was well below 100%. But then, a practical CELSS on the Moon might also depend on regular resupply from Earth. Several important lessons have been learned from the CELSS research described in this review.

Review↗
NASA NTRS1973

Explosives and pyrotechnic propellants for use in long term deep space missions

Explosives and pyrotechnic propellant materials which will withstand heat sterilization cycling at 125 C and ten year deep space aging under 10 to the minus 6th power torr and 66 C have been selected. The selection was accomplished through a detailed literature survey and an analytical evaluation of the physicochemical properties of the materials. The chemical components of the electroexplosive devices used in U.S. missiles and spacecraft were categorized into primary explosives, secondary explosives, and propellant ingredients. Kinetic data on such parameters as thermal decomposition and sublimation were obtained for these materials and used as a basis for the ten year life prediction. From these experimental data and some analytical calculations, a listing of candidate materials for deep space missions was made.

Gorzynski, C. S., Jr.↗
NASA NTRSDate not supplied

Preparation for and Performance of a Pseudomonas aeruginosa Biofilm Experiment On Board the International Space Station

Biofilms are a problem on Earth given their ability to degrade the materials upon which they grow and due to their relevance to infections. Remarkably, 65% and 80% of infections and chronic diseases on Earth are associated with biofilms, respectively. In space, these problems' impact is higher because the crew's lives and mission success depend on nominal operation of mechanical systems. Furthermore, the isolated confined environment nature of spaceflight may increase the rates of disease transmission. In the case of the International Space Station (ISS), biofilms are an identified problem on the Environmental Control and Life Support System (ECLSS), namely on the water processor assembly (WPA). In late 2019, the Space Biofilms experiment launched towards ISS to (i) characterize the mass, thickness, morphology, and gene expression of biofilms formed in space with respect to matched Earth controls, (ii) interrogate the expression of antimicrobial resistance genes, and (iii) test novel materials as potential biofilm control strategies for future ECLSS components. For this, 288 bacterial samples were prepared prior to the launch of the Northrop Grumman CRS-12 mission from NASA's Wallops Flight Facility. The samples were integrated into the spaceflight hardware, BioServe's Fluid Processing Apparatus (FPA) packed in sets of eight in Group Activation Packs (GAP). Half of these samples were activated and terminated on orbit by NASA astronauts Jessica Meir and Christina Koch, while the remaining half were processed equivalently on Earth. The spaceflight bacterial samples of Space Biofilms returned on board SpaceX' CRS-19 Dragon spacecraft, in early 2020. We here describe the test campaign implemented to verify the experiment design and confirm it would enable us to achieve the project's scientific goals. This campaign ended with the Experiment Verification Test (EVT), from which we here present example morphology and transcriptomic results. We describe in detail the sample preparation prior to flight, including cleaning and sterilization of the coupons of six materials (SS316, passivated-SS316, lubricant impregnated surface, catheter-grade silicone with and without a nanotopography, and cellulose membrane), loading and integration of growth media, bacterial inoculum, fixative and preservative to enable experiment termination on orbit. Additionally, we describe the performance of the experiment on board the ISS, including crew activities, use of assets, temperature profile, and experiment timeline; all leading to a successful spaceflight experiment.

Pamela Flores↗
NASA NTRS2005

NASA Tech Briefs, October 2005

Topics covered include: Insect-Inspired Optical-Flow Navigation Sensors; Chemical Sensors Based on Optical Ring Resonators; A Broad-Band Phase-Contrast Wave-Front Sensor; Progress in Insect-Inspired Optical Navigation Sensors; Portable Airborne Laser System Measures Forest-Canopy Height; Deployable Wide-Aperture Array Antennas; Faster Evolution of More Multifunctional Logic Circuits; Video-Camera-Based Position-Measuring System; N-Type delta Doping of High-Purity Silicon Imaging Arrays; Avionics System Architecture Tool; Updated Chemical Kinetics and Sensitivity Analysis Code; Predicting Flutter and Forced Response in Turbomachinery; Upgrades of Two Computer Codes for Analysis of Turbomachinery; Program Facilitates CMMI Appraisals; Grid Visualization Tool; Program Computes Sound Pressures at Rocket Launches; Solar-System Ephemeris Toolbox; Data-Acquisition Software for PSP/TSP Wind-Tunnel Cameras; Corrosion-Prevention Capabilities of a Water-Borne, Silicone-Based, Primerless Coating; Sol-Gel Process for Making Pt-Ru Fuel-Cell Catalysts; Making Activated Carbon for Storing Gas; System Regulates the Water Contents of Fuel-Cell Streams; Five-Axis, Three-Magnetic-Bearing Dynamic Spin Rig; Modifications of Fabrication of Vibratory Microgyroscopes; Chamber for Growing and Observing Fungi; Electroporation System for Sterilizing Water; Thermoelectric Air/Soil Energy-Harvesting Device; Flexible Metal-Fabric Radiators; Actuated Hybrid Mirror Telescope; Optical Design of an Optical Communications Terminal; Algorithm for Identifying Erroneous Rain-Gauge Readings; Condition Assessment and End-of-Life Prediction System for Electric Machines and Their Loads; Lightweight Thermal Insulation for a Liquid-Oxygen Tank; Stellar Gyroscope for Determining Attitude of a Spacecraft; and Lifting Mechanism for the Mars Explorer Rover.

Source record↗
NASA NTRS1974

Development of the Viking Mars lander thermal control subsystem design

Two Viking spacecraft, each consisting of a lander capsule coupled to an orbiter, will be launched toward Mars during the summer of 1975. About a year later, the orbiters will go into orbit around Mars and the landers will descend to the surface for 90-day landed missions. The lander must withstand a wide variety of environmental and operational conditions during all phases of the mission, including prelaunch sterilization. On the surface of Mars, the lander internal temperatures must be controlled under widely varying thermal environments and atmospheric conditions. The lander thermal design is based on the maximum use of passive techniques and is integrated into the overall vehicle design and operation. The solutions to the unusual combinations of design problems and a summary of the results of full scale model testing under simulated mission conditions are presented.

Morey, T. F.↗
NASA NTRS2012

Survival of B. Horneckiae Spores Under Ground-simulated Space Conditions

To prevent forward contamination and maintain the scientific integrity of future life detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated habitats, spore-forming microbes are highly resistant to various physical and chemical conditions, which include ionizing and UV radiation, desiccation and oxidative stress, and the harsh environment of outer space or planetary surfaces. Recently a radiation resistant, spore forming bacterial isolate, Bacillus horneckiae, was isolated from a clean room of the Kennedy Space Center where the Phoenix spacecraft was assembled. The exceptionally high tolerance of extreme conditions demonstrated by sporeforming bacteria highlighted the need to assess the viability of these microbes in situ (in real) space. The proposed BOSS (Biofilm Organisms Surfing Space) project aims to understand the mechanisms by which biofilm forming organisms, such as B. horneckiae, will potentially be able to withstand harsh space conditions. As previously stated, the spore producing ability of these species gives them increased survivability to harsh conditions. Some of the spores will have the protective exosporium layer artificially removed before the test to determine if the existence of this layer significantly changes the survivability during the mission. In preparation for that experiment, we analyzed spores which were exposed during a ground simulation, the EXPOSE R2 Biofilm Organisms Surfing Space (BOSS). Previous to exposure, spores were deposited onto spacecraft grade aluminum coupons in a spore suspension calculated to contain between 10(exp 7) and 10(exp 8) spores. This precursor series will be used to establish a baseline survivability function for comparison with the future flight tests during EXPOSE-R. For each coupon, a 10% polyvinyl alcohol (PVA) film was applied and peeled from the coupon to recover the spores. One hundred μl of sterile 10% PVA was applied to the surface of the coupon and allowed to dry for 1 hour at 37 C. The films were then removed using sterile scalpel and forceps and placed into a glass test tube containing 2 milliliters of sterile deionized water. The PVA film process was then repeated on each coupon one additional time to ensure recovery of the majority of spores. The second PVA film was added in the same glass tube as in the previous round. If the spores remained 100% viable, the test tubes should now contain between 5 X 10(exp 6) and 5 X 10(exp 7) spores per millimeter; however, it is expected that some loss of viability has occurred. In order to assess this loss, the number of colony forming, viable spores was counted. To count the colony forming units (CFUs), the spore containing solution was diluted in a process of 10-fold serial dilution by mixing successive solutions in a 100 microliter spore suspension to 900 microliter deionized H2O ratio. A sample dilution series revealed that 10(exp -3) and 10(exp -4) concentrations would be necessary for an accurate CFU count to be taken. For those two concentrations, a spread on a TSA plate was prepared and incubated at 32 C. For the samples exposed to UV radiation, the cell survivability was too low to establish a count from 100 microliter spread plating. Instead, no dilutions were performed and the entire 2 milliliter spore suspension was plated and incubated at 32 C. The plate's CFU counts were taken at 24 hours and 48 hours from the time of plating. At the end of the CFU counting the total surviving spores in each sample were calculated based on the number of CFUs that were observed per 100 microliters, or per 2 milliliters for the UV irradiated samples. The results of these calculations are shown in Figures 1 and 2.

cleanroom↗
NASA NTRS2009

Efficacy of Various Chemical Disinfectants on Biofilms Formed in Spacecraft Potable Water System Component

As the provision of potable water is critical for successful habitation of the International Space Station (ISS), life support systems were installed in December 2008 to recycle both humidity from the atmosphere and urine to conserve available water in the vehicle. Pre-consumption testing from the dispensing needle at the Potable Water Dispenser (PWD) indicated that bacterial concentrations exceeded the current ISS specifications of 50 colony forming units (CFU) per ml. Subsequent investigations revealed that a corrugated stainless steel flex hose upstream of the dispensing needle in the PWD was filled with non-sterile water and left at room temperature for over one month before launch. To simulate biofilm formation that was suspected in the flight system, sterile flex hoses were seeded with a consortium of bacterial isolates previously recovered from other ISS water systems, which included Ralstonia pickettii, Burkholderia multivorans, Caulobacter vibrioides., and Cupriavidus pauculus. After 5 days of incubation, these hoses were challenged with various chemical disinfectants including hydrogen peroxide, colloidal silver, and buffered pH solutions to determine the ability of the disinfectants to decrease and maintain bacterial concentrations below ISS specifications. Disinfection efficacy over time was measured by collecting daily heterotrophic plate counts following exposure to the disinfectants. A single flush with either 6% hydrogen peroxide solution or a mixture of 3% hydrogen peroxide and 400 ppb colloidal silver effectively reduced the bacterial concentrations to less than 1 CFU/ml for a period of up to 2 months. Testing results indicated that hydrogen peroxide and mixtures of hydrogen peroxide and colloidal silver have tremendous potential as alternative disinfectants for ISS water systems.

Wong, Willy↗
NASA NTRS2018

A Prototype Tool for Assessing Forward Contamination from Space Suits

Crewed exploration missions dramatically increase the capability for large-scale sample collection and return activities, but they also increase the possibility and likelihood of forward contamination. Systematic research on forward contamination from uncrewed spacecraft has steadily progressed since the Viking missions, but parallel research on contamination from space suits has not. Current space suits have leakage rates as high as 100 cc gas/min., but it is unclear how many or what types of microbes are exiting the suits along with this gas. The Human Forward Contamination Assessment team at NASA's Johnson Space Center (JSC) has developed a prototype swab tool that is capable of maintaining sterility during pressure changes associated with entering and exiting vacuum. The primary objective of recent Extravehicular Activity (EVA) Swab Kit testing is to characterize the type of microorganisms typically found on spacesuit external surfaces under suit differential pressure conditions. Most human-associated microorganisms can fit through a 0.5 to 1.0 μm gap. Understanding potential leak paths will inform future hardware design decisions. Knowing which types of microorganisms may leak from EVA suits provides a basis for subsequent studies to characterize the viability of those organisms under destination conditions, as well as how far they might spread through natural or human-influenced processes. The results of EVA suit molecular microbial community analyses will inform NASA exploration mission operations and hardware design, and help close Strategic Knowledge Gap B5, Forward Contamination to Mars.

Rucker, M.↗