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NASA NTRS2006

Use of Atmospheric Glow Discharge Plasma to Modify Spaceport Materials

Numerous materials used in spaceport operations require stringent evaluation before they can be utilized. It is critical for insulative polymeric materials that any surface charge be dissipated as rapidly as possible to avoid Electrostatic Discharges (ESD) that could present a danger. All materials must pass the Kennedy Space Center (KSC) standard electrostatic test [1]; however several materials that are considered favorable for Space Shuttle and International Space Station use have failed. Moreover, to minimize contamination of Mars spacecraft, spacecraft are assembled under cleanroom conditions and specific cleaning and sterilizing procedures are required for all materials. However, surface characteristics of these materials may allow microbes to survive by protecting them from sterilization and cleaning techniques. In this study, an Atmospheric Pressure Glow Discharge Plasma (APGD) [2] was used to modify the surface of several materials. This allowed the materials surface to be modified in terms of hydrophilicity, roughness, and conductivity without affecting the bulk properties. The objectives of this study were to alter the surface properties of polymers for improved electrostatic dissipation characteristics, and to determine whether the consequent surface modification on spaceport materials enhanced or diminished microbial survival.

Trigwell, S.↗

NASA NTRS2013

Intravenous Fluid Generation System

The ability to stabilize and treat patients on exploration missions will depend on access to needed consumables. Intravenous (IV) fluids have been identified as required consumables. A review of the Space Medicine Exploration Medical Condition List (SMEMCL) lists over 400 medical conditions that could present and require treatment during ISS missions. The Intravenous Fluid Generation System (IVGEN) technology provides the scalable capability to generate IV fluids from indigenous water supplies. It meets USP (U.S. Pharmacopeia) standards. This capability was performed using potable water from the ISS; water from more extreme environments would need preconditioning. The key advantage is the ability to filter mass and volume, providing the equivalent amount of IV fluid: this is critical for remote operations or resource- poor environments. The IVGEN technology purifies drinking water, mixes it with salt, and transfers it to a suitable bag to deliver a sterile normal saline solution. Operational constraints such as mass limitations and lack of refrigeration may limit the type and volume of such fluids that can be carried onboard the spacecraft. In addition, most medical fluids have a shelf life that is shorter than some mission durations. Consequently, the objective of the IVGEN experiment was to develop, design, and validate the necessary methodology to purify spacecraft potable water into a normal saline solution, thus reducing the amount of IV fluids that are included in the launch manifest. As currently conceived, an IVGEN system for a space exploration mission would consist of an accumulator, a purifier, a mixing assembly, a salt bag, and a sterile bag. The accumulator is used to transfer a measured amount of drinking water from the spacecraft to the purifier. The purifier uses filters to separate any air bubbles that may have gotten trapped during the drinking water transfer from flowing through a high-quality deionizing cartridge that removes the impurities in the water before entering the salt bag and mixing with the salt to create a normal saline solution.

McQuillen, John↗

NASA NTRS2013

Microbiological Sampling Methods and Sanitation of Edible Plants Grown on ISS

Pathogenic microbes on the surfaces of salad crops and growth chambers pose a threat to the health of crew on International Space Station. For astronauts to safely consume spacegrown vegetables produced in NASA's new vegetable production unit, VEGGIE, three technical challenges must be overcome: real-time sampling, microbiological analysis, and sanitation. Raphanus sativus cultivar Cherry Bomb II and Latuca sativa cultivar Outredgeous, two saled crops to be grown in VEGGIE, were inoculated with Salmonella enterica serovar Typhimurium (S. Typhimurium), a bacterium known to cause food-borne illness~ Tape- and swab-based sampling techniques were optimized for use in microgravity and assessed for effectiveness in recovery of bacteria from crop surfaces: Rapid pathogen detection and molecular analyses were performed via quantitative real-time polymerase chain reactiop using LightCycler® 480 and RAZOR® EX, a scaled-down instrument that is undergoing evaluation and testing for future flight hardware. These methods were compared with conventional, culture-based methods for the recovery of S. Typhimurium colonies. A sterile wipe saturated with a citric acid-based, food-grade sanitizer was applied to two different surface materials used in VEGGIE flight hardware that had been contaminated with the bacterium Pseudomonas aeruginosa,. another known human pathogen. To sanitize surfaces, wipes were saturated with either the sanitizer or sterile deionized water and applied to each surface. Colony forming units of P. aeruginosa grown on tryptic soy agar plates were enumerated from surface samples after sanitization treatments. Depending on the VEGGIE hardware material, 2- to 4.5-log10 reductions in colony-forming units were observed after sanitization. The difference in recovery of S. Typhimurium between tape- and swab- based sampling techniques was insignificant. RAZOR® EX rapidly detected S. Typhimurium present in both raw culture and extracted DNA samples.

Parrish, Charles H. II↗

NASA NTRS2014

IVGEN Post Flight Analysis

The Intravenous Fluid Generation (IVGEN) Experiment was a technology demonstration experiment that purified ISS potable water, mixed it with salt, and transferred it through a sterilizing filter. On-orbit performance was verified as appropriate and two 1.5 l bags of normal saline solution were returned to earth for post-flight testing by a FDA certified laboratory for compliance with United States Pharmacopiea (USP) standards. Salt concentration deviated from required values and an analysis identified probable causes. Current efforts are focused on Total Organic Content (TOC) testing, and shelf life.The Intravenous Fluid Generation (IVGEN) Experiment demonstrated the purification of ISS potable water, the mixing of the purified water with sodium chloride, and sterilization of the solution via membrane filtration. On-orbit performance was monitored where feasible and two 1.5-liter bags of normal saline solution were returned to earth for post-flight testing by a FDA-registered laboratory for compliance with United States Pharmacopeia (USP)standards [1]. Current efforts have been focused on challenge testing with identified [2] impurities (total organic-carbon), and shelf life testing. The challenge testing flowed known concentrations of contaminants through the IVGEN deionizing cartridge and membrane filters to test their effectiveness. One finding was that the filters and DI-resin themselves contribute to the contaminant load during initial startup, suggesting that the first 100 ml of fluid be discarded. Shelf life testing is ongoing and involves periodic testing of stored DI cartridges and membrane filters that are capped and sealed in hermetic packages. The testing is conducted at six month intervals measuring conductivity and endotoxins in the effluent. Currently, the packaging technique has been successfully demonstrated for one year of storage testing. The USP standards specifies that the TOC be conducted at point of generation as opposed to point of consumption. Samples were generated and shipped to an FDA facility however, testing determined that the samples failed the TOC specification with most likely due to leaching from the sample container. Shelf life testing is examining packaging techniques and periodic testing of samples of DI cartridges that are capped and sealed in hermetic packages. Periodic testing of the purified water output will be conducted at six month intervals.

microgravity applications↗

NASA NTRS2015

Alternate Protocol for Detecting Biological Contamination on Sensitive Hardware

The purpose of this project is to develop a sterile water based rapid bioburden test. Contamination engineers use two tests to assess the level of biological contamination on hardware: the rapid five minute bioburden test, which is a molecular screening for Adenosine triphosphate (ATP), a molecule found in all cells on the hardware, and a slower colony growth test, which is used to give a more accurate representation of the amount of microbes on the hardware. However, the rapid bioburden test has limited application because it leaves a residue that can be detrimental to sensitive hardware. This can cause project delays while waiting for the results from the three day colony growth test. We address this problem by adapting the commercial germicide based ATP system to a sterile water based system. The test works by reacting ATP with D-Luciferin and Luciferase protein to yield light. The light is then detected by a luminometer that outputs a Relative Light Unit (RLU) amount depending on how much ATP is present. To analyze the effectiveness of the new test, we developed a correlation between amounts of ATP and the RLU produced using the germicide based system. From these experiments, we've generated a consistent relationship between the two in the form of a power curve. From there, we developed a correlation curve between the amount of colonies and the RLU they produced. Initial tests of the new protocol have shown that the water based system isn't as sensitive as the germicide based test.

planetary protection↗

NASA NTRS2016

A Carbonaceous Chondrite Based Simulant of Phobos

In support of an ESA-funded concept study considering a sample return mission, a simulant of the Martian moon Phobos was needed. There are no samples of the Phobos regolith, therefore none of the four characteristics normally used to design a simulant are explicitly known for Phobos. Because of this, specifications for a Phobos simulant were based on spectroscopy, other remote measurements, and judgment. A composition based on the Tagish Lake meteorite was assumed. The requirement that sterility be achieved, especially given the required organic content, was unusual and problematic. The final design mixed JSC-1A, antigorite, pseudo-agglutinates and gilsonite. Sterility was achieved by radiation in a commercial facility.

simulant↗

NASA NTRS2018

EVA Swab Tool to Support Planetary Protection and Astrobiology Evaluations

When we send humans to search for life on other planets, we'll need to know what we brought with us versus what may already be there. To ensure our crewed systems meet planetary protection requirements-and to protect our science from human contamination-we'll need to assess whether microorganisms may be leaking or venting from our spacecraft. Microbial sample collection outside of a pressurized spacecraft is complicated by temperature extremes, low pressures that preclude the use of laboratory standard (wetted) swabs, and operation either in bulky spacesuits or with robotic assistance. Engineers at the National Aeronautics and Space Administration (NASA) recently developed a swab kit for use in collecting microbial samples from the external surfaces of crewed spacecraft, including spacesuits. The Extravehicular Activity (EVA) Swab Kit consists of a single swab tool handle and an eight-canister sample caddy. The design team minimized development cost by re-purposing a heritage Space Shuttle tile repair handle that was designed to quickly snap into different tool attachments by engaging a mating device in each attachment. This allowed the tool handle to snap onto a fresh swab attachment much like popular shaving razor handles can snap onto a disposable blade cartridge. To disengage the handle from a swab, the user performs two independent functions, which can be done with a single hand. This dual operation mitigates the risk that a swab will be inadvertently released and lost in microgravity. Each swab attachment is fitted with commercially available foam swab tips, vendor-certified to be sterile for Deoxyribonucleic Acid (DNA). A microbial filter installed in the bottom of each sample container allows the container to outgas and repressurize without introducing microbial contaminants to internal void spaces. Extensive ground testing, post-test handling, and sample analysis confirmed the design is able to maintain sterile conditions as the canister moves between various pressure environments. To further minimize cost, the design team acquired extensive ground test experience in a relevant flight environment by piggy-backing onto suited crew training runs. These training runs allowed the project to validate tool interfaces with pressurized EVA gloves and collect user feedback on the tool design and function, as well as characterize baseline microbial data for different types of spacesuits. In general, test subjects found the EVA Swab Kit relatively straightforward to operate, but identified a number of design improvements that will be incorporated into the final design. Although originally intended to help characterize human forward contaminants, this tool has other potential applications, such as for collecting and preserving space-exposed materials to support astrobiology experiments.

Rucker, Michelle A.↗

NASA NTRS2017

Evaluation of Low-Pressure Cold Plasma for Disinfection for ISS Grown Produce and Metallic Instrumentation

Cold plasma (CP) cleaning is a dry, non-thermal process, which can provide broad-spectrum antimicrobial activity yet reportedly causes little to no damage to the object being sanitized. Since cold plasma uses no liquids, it has the distinct advantage when used in microgravity of not having to separate liquids from the item being cleaned. This paper will present results on an effort to use low pressure CP to disinfect or sterilize materials for in space applications. Exposure times from 0 to 60 minutes and pressures ranging from 10 to 100 Pa were used to optimize plasma parameters to achieve acceptable kill rates for 3 bacteria, Bacillus cereus, E. coli and Bacillus pumulis SAFR-32 and one fungi, Aspergillus niger. These tests were done on produce and metal coupons to simulate medical equipment. Produce testing was not successful, with unacceptable kill rates and the produce being negatively impacted by exposure to the plasma. The plasma caused a 5 log reduction in the number of viable bacteria on metal coupon tests, which placed the number of viable bacteria below the detection limit. This is a very promising result showing that sterilization of medical equipment with cold plasma is feasible. Scanning Electron Microscope images were taken before and after exposure. The images after plasma exposure shows that the bacteria spores have been physically affected, as their size has gotten smaller and the appearance has changed.

Hintze, Paul E.↗

NASA NTRS2017

Evaluation of Low-Pressure Cold Plasma for Disinfection of ISS Grown Produce and Metallic Instrumentation

Cold plasma (CP) cleaning is a dry, non-thermal process, which can provide broad-spectrum antimicrobial activity yet reportedly causes little to no damage to the object being sanitized. Since cold plasma uses no liquids, it has the distinct advantage when used in microgravity of not having to separate liquids from the item being cleaned. This paper will present results on an effort to use low pressure CP to disinfect or sterilize materials for in space applications. Exposure times from 0 to 60 minutes and pressures ranging from 0.10 to 1.0 mbar were used to optimize plasma parameters. Tests were done on produce and metal coupons to simulate medical equipment. Escherichia coli was used as the challenge organism on produce and Bacillus pumilus SAFR-32 was used on metal surfaces. Produce testing was not successful, with unacceptable kill rates and the produce being negatively impacted by exposure to the plasma. The plasma caused a 5 log reduction in the number of viable bacteria on metal coupon tests, which placed the number of viable bacteria below the detection limit. This is a very promising result showing that sterilization of medical equipment with cold plasma is feasible. Scanning Electron Microscope images were taken before and after exposure. The images after plasma exposure show that the bacteria spores have been physically affected, as their size has gotten smaller and their appearance has changed.

Hintze, Paul E.↗

NASA NTRS2018

EVA Swab Tool to Support Planetary Protection and Astrobiology Evaluations

When we send humans to search for life on other planets, we'll need to know what we brought with us versus what may already be there. To ensure our crewed systems meet planetary protection requirements-and to protect our science from human contamination-we'll need to assess whether microorganisms may be leaking or venting from our spacecraft. Microbial sample collection outside of a pressurized spacecraft is complicated by temperature extremes, low pressures that preclude the use of laboratory standard (wetted) swabs, and operation either in bulky spacesuits or with robotic assistance. A team at the National Aeronautics and Space Administration (NASA) recently developed a swab kit for use in collecting microbial samples from the external surfaces of crewed spacecraft, including spacesuits. The Extravehicular Activity (EVA) Swab Kit consists of a single swab tool handle and an eight-canister sample caddy. The design team minimized development cost by re-purposing a heritage Space Shuttle tile repair handle that was designed to quickly snap into different tool attachments by engaging a mating device in each end effector. This allowed the tool handle to snap onto a fresh swab end effector much like popular shaving razor handles can snap onto a disposable blade cartridge. To disengage the handle from a swab, the user performs two independent functions, which can be done with a single hand. This dual operation mitigates the risk that a swab will be inadvertently released and lost in microgravity. Each swab end effector is fitted with commercially available foam swab tips, vendor-certified to be sterile for Deoxyribonucleic Acid (DNA). A microbial filter installed in the bottom of each sample container allows the container to outgas and re-pressurize without introducing microbial contaminants to internal void spaces. Extensive ground testing, post-test handling, and sample analysis confirmed the design is able to maintain sterile conditions as the canister moves between various pressure environments. To further minimize cost, the design team acquired extensive ground test experience in a relevant flight environment by piggy-backing onto suited crew training runs. These training runs allowed the project to validate tool interfaces with pressurized EVA gloves and collect user feedback on the tool design and function, as well as characterize baseline microbial data for different types of spacesuits. In general, test subjects found the EVA Swab Kit relatively straightforward to operate, but identified a number of design improvements that will be incorporated into the final design. Although originally intended to help characterize human forward contaminants, this tool has other potential applications, such as for collecting and preserving space-exposed materials to support astrobiology experiments.

Rucker, Michelle A.↗

NASA NTRS2018

Effect of Ex Vivo Ionizing Radiation on Static and Fatigue Properties of Mouse Vertebral Bodies

For a variety of medical and scientific reasons, human bones can be exposed to a wide range of ionizing radiation levels. In vivo radiation therapy (0.05 kGy) is used in cancer treatment, and ex vivo irradiation (25-35 kGy) is used to sterilize bone allografts. Ionizing radiation in these applications has been shown to increase risk of fracture, decrease bone quality and degrade collagen integrity. Past studies have investigated the deleterious effects of radiation on cortical or trabecular bone specimens individually, but to date no studies have examined whole bones containing both cortical and trabecular tissue. Furthermore, a clear relationship between the dose and the mechanical and biochemical response of bone's extracellular matrix has yet to be established for doses ranging from cancer therapy to allograft sterilization (0.05-35 kGy). To gain insight into these issues, we conducted an ex vivo radiation study to investigate non-cellular (i.e. matrix) effects of ionizing radiation dose on vertebral whole bone mechanical properties, over a range of radiation doses (0.05-35 kGy), with a focus on any radiation-induced changes in collagen. With underlying mechanisms of action in mind, we hypothesized that any induced reductions in mechanical properties would be associated with changes in collagen integrity. METHODS: 20-week old female mice were euthanized and the lumbar spine was dissected using IACUC approved protocols. The lumbar vertebrae (L1- S1) were extracted from the spine via cuts through adjacent intervertebral discs, and the endplates, posterior processes, surrounding musculature, and soft tissues were removed (approx. 1.5mm diameter, approx. 2mm height). Specimens were randomly assigned to one of five groups for ex vivo radiation exposure: x-ray irradiation at 0.05, 1, 17, or 35 kGy, or a 0 kGy control. Following irradiation, the vertebrae were imaged using microcomputed tomography (micro-CT) and then subjected to either monotonic compressive loading to failure or uniform cyclic compressive loading. During cyclic testing, samples were loaded in force control to a force level that corresponded to a strain of 0.46%, as determined in advance by a linearly elastic micro-CT-based finite element analysis for each specimen. Tests were stopped at imminent fracture, defined as a rapid increase in strain. The main outcome for the monotonic test was the strength (maximum force); for cyclic testing it was the fatigue life (log of the number of cycles of loading at imminent failure). A fluorometric assay was used on the S1 vertebrae to measure the number of non-enzymatic collagen crosslinks[4]. A one-way ANOVA was performed on mechanical properties and collagen crosslinks; means were compared with controls using Dunnett's method, with a Tukey-Kramer post-hoc analysis when significance was found (p < or = 0.05). RESULTS: Compared to the unirradiated control group, the concentration of non-enzymatic collagen crosslinks was significantly increased for all irradiated groups (p < 0.0001), and being higher by at least 50% (Figure 1a). By contrast, the radiation effects on the collagen were only evident at the higher doses. For irradiation exposures of 17 kGy or more, strength decreased substantially as the radiation level was increased, but no effect was evident below 17 kGy (Figure 1b). There was no significant change in the stiffness or maximum displacement for any radiation dose (p>0.05). The finite element analysis prescribed force level for cyclic loading exceeded the measured (monotonic) strength of the 17 and 35 kGy irradiated groups (mean +/- SD, 20.6 +/- 5.6 N; 13.2 +/- 3.7 N, respectively) and therefore these groups were eliminated from the fatigue study. The fatigue life for the 0.05 and 1 kGy groups were similar to each other and were not statistically significantly different from the control group (Figure 1c).

radiation↗

NASA NTRS2018

EVA Swab Tool to Support Planetary Protection and Astrobiology Evaluations

When we send humans to search for life on other planets, we'll need to know what we brought with us versus what may already be there. To ensure our crewed systems meet planetary protection requirements?and to protect our science from human contamination?we'll need to assess whether microorganisms may be leaking or venting from our spacecraft. Microbial sample collection outside of a pressurized spacecraft is complicated by temperature extremes, low pressures that preclude the use of laboratory standard (wetted) swabs, and operation either in bulky spacesuits or with robotic assistance. A team at the National Aeronautics and Space Administration (NASA) recently developed a swab kit for use in collecting microbial samples from the external surfaces of crewed spacecraft, including spacesuits. The Extravehicular Activity (EVA) Swab Kit consists of a single swab tool handle and an eight-canister sample caddy. The design team minimized development cost by re-purposing a heritage Space Shuttle tile repair handle that was designed to quickly snap into different tool attachments by engaging a mating device in each end effector. This allowed the tool handle to snap onto a fresh swab end effector much like popular shaving razor handles can snap onto a disposable blade cartridge. To disengage the handle from a swab, the user performs two independent functions, which can be done with a single hand. This dual operation mitigates the risk that a swab will be inadvertently released and lost in microgravity. Each swab end effector is fitted with commercially available foam swab tips, vendor-certified to be sterile for Deoxyribonucleic Acid (DNA). A microbial filter installed in the bottom of each sample container allows the container to outgas and re-pressurize without introducing microbial contaminants to internal void spaces. Extensive ground testing, post-test handling, and sample analysis confirmed the design is able to maintain sterile conditions as the canister moves between various pressure environments. To further minimize cost, the design team acquired extensive ground test experience in a relevant flight environment by piggy-backing onto suited crew training runs. These training runs allowed the project to validate tool interfaces with pressurized EVA gloves and collect user feedback on the tool design and function, as well as characterize baseline microbial data for different types of spacesuits. In general, test subjects found the EVA Swab Kit relatively straightforward to operate, but identified a number of design improvements that will be incorporated into the final design. Although originally intended to help characterize human forward contaminants, this tool has other potential applications, such as for collecting and preserving space-exposed materials to support astrobiology experiments.

planetary protection↗

NASA NTRS2018

Microbial Ecology of NASA Curation Clean Rooms

Clean room standards like ISO 14644 used for facilities that construct spacecraft and store returned samples do not explicitly account for microbial contamination. While there are associated ISO standards for monitoring and controlling bio-contamination in clean rooms it is not always standard practice to do so. The NASA Astromaterials Acquisition and Curation Office maintains seven separate clean labs for storing extraterrestrial samples from the Moon, meteorites, cosmic dust, asteroids, comets, solar wind particles, and microparticle impact samples. These labs are routinely monitored for particulate and trace metal contamination. However, the sample collections are either non-sterile at the time of collection (e.g., meteorites) or are no longer being used to address scientific questions that could be affected by non-sterile conditions (e.g., Lunar samples). Outside of isolated studies there has not been a systematic, longitudinal characterization of the microbial ecology of NASA curation clean rooms. In accordance with the advanced curation initiative, and to prepare for future sample return missions, we have initiated a routine microbiological monitoring program in the Antarctic Meteorite Lab. This monitoring program will be used to determine what microbes are capable of surviving in these oligotrophic environments and whether or not they are capable of altering the sample collections in any significant manner. Repeat sampling will allow us to understand how routine use of these labs affects the microbial ecology over time.

Regberg, A. B.↗

NASA NTRS2017

Heating and Thermal Control of Brazing Technique to Break Contamination Path for Potential Mars Sample Return

The potential return of Mars sample material is of great interest to the planetary science community, as it would enable extensive analysis of samples with highly sensitive laboratory instruments. It is important to make sure such a mission concept would not bring any living microbes, which may possibly exist on Mars, back to Earth’s environment. In order to ensure the isolation of Mars microbes from Earth’s Atmosphere, a brazing sealing and sterilizing technique was proposed to break the Mars-to-Earth contamination path. Effectively, heating the brazing zone in high vacuum space and controlling the sample temperature for integrity are key challenges to the implementation of this technique. The break-the-chain procedures for container configurations, which are being considered, were simulated by multi-physics finite element models. Different heating methods including induction and resistive/radiation were evaluated. The temperature profiles of Martian samples in a proposed container structure were predicted. The results show that the sealing and sterilizing process can be controlled such that the samples temperature is maintained below the level that may cause damage, and that the brazing technique is a feasible approach to breaking the contamination path.

Bao, Xiaoqi↗

NASA NTRSDate not supplied

ISS External Microorganisms: A Payload to Close Planetary Protection Knowledge Gaps for Crewed Missions

Before NASA or COSPAR is able to set planetary protection requirements for crewed missions to locations like Mars there are a number of critical knowledge gaps that must be addressed (1). One of the most important knowledge gaps is an understanding of microbial leakage from crewed habitats and space suits. Current ECLSS (Environmental Control and Life Support System) and PLSS (Portable Life Support System) requirements do not include any provisions to control microbes that may escape along with vented or leaked gasses. The current generation of NASA space suits can leak at rates as high as 100 cm2 /min. during nominal operation (2). ISS (International Space Station) intentionally vents atmospheric gases like CO2 to maintain habitable conditions for the crew. Furthermore, every time an airlock is used for EVA (extravehicular activity)there is an accompanying release of internal atmosphere. Since it is not possible to sterilize a crewed mission, it is important that we understand what if any microbes are entrained in these vented and leaked products. It is also important to understand if these microbes can survive on exterior surfaces. Recent sampling of the Russian segments of ISS suggest that bacteria and fungi from inside ISS may be capable of surviving on external surfaces(3). NASA is developing an aseptic sampling tool for use during EVA and plans to collect samples from vents on ISS to build on these results. The results of this work will be used to develop planetary protection requirements for vented and leaked gasses from crewed volumes. NASA has developed and tested a tool kit for collecting microbiological samples during EVA(4). This tool kit contains eight commercially available, 23 mm. diameter, foam swabs that can be used to aseptically collect samples while at vacuum. The swabs are individually housed in aluminum canisters that are equipped with 0.2 μm Teflon filters. These filters allow the canisters to equilibrate to pressure changes while preventing microbiological contamination. The canisters will be cleaned and sterilized before flight. Results from ground-based testing indicate that this tool kit is capable of aseptically collecting microbes while at vacuum without becoming contaminated during pressure changes(5). Based on the results of this ground testing we have modified the tool kit to meet NASA safety requirements and improve the ergonomics. We added additional mounting points to the tool kit to give astronauts more options for securing it during use. We also changed the opening mechanism to improve the precision with which swabs can be extracted from the tool kit. We plan to use this kit on an upcoming EVA to collect samples from non-propulsive vents and areas near the U.S. airlock on ISS. These samples will be frozen at -80 ̊C and stored on station until they can be returned to Earth. We will analyze these returned samples using next generation DNA sequencing to determine the community composition and function of external ISS environments. The results of this study will close planetary protection knowledge gaps for crewed missions and will help NASA determine appropriate planetary protection requirements for life support systems. The tool kit will also be useful for collecting aseptic samples on upcoming crewed or robotic missions and could easily be modified to collect samples with organic contamination control requirements as well.

A B Regberg↗

NASA NTRSDate not supplied

Mars Sample Receiving Facility Research and Development to Enable Preservation, Safe Containment, and Scientific Research of Martian Samples on Earth

NASA and ESA are working together to plan a joint campaign to potentially bring back the first Martian samples to Earth in the early 2030s. On the surface of Mars, the Mars 2020Perseverance Rover is selecting and packaging samples that could be returned to Earth for careful examination by an international team of scientists. In preparation for this historic sample return, advance planning is underway to design and build a Mars Sample Receiving Facility (SRF) in the mid-2020s. In 2022, a Mars Sample Receiving Facility Assessment Study(MSAS) will be conducted to further define construction modality and capability options for a facility in the conterminous United States. Since Mars Sample Return (MSR) is currently categorized as a planetary protection restricted Earth return, this new facility would feature biosafety level 4 (BSL-4)-like high containment to protect Earth’s biosphere from any potential hazard. In addition, the facility also requires integration of cleanroom technologies to mitigate against terrestrial contamination and preserve sample integrity for science investigations. The integration of both clean handling and high containment requires significant technology research and development (R&D) by the mid-2020s to support the definition and planning of a SRF before commencing the site specific design phase. The SRF project will need to solidify the inorganic, organic, and biological contamination control (CC) requirements for the facility. This could entail a translation of Mars 2020 mission CC requirements for geologic material to engineering requirements for surfaces and airborne molecular contamination inside isolators and cleanrooms. In tandem, the project will need to determine precision cleaning and sterilization approaches for cleanrooms, isolators, equipment, and tools. Initial material selections for the facility construction materials, cleanrooms, isolators, and BSL-4 suits/garments are an important activity. The recent construction of the OSIRIS-REx and Hayabusa2 curation cleanroom laboratories greatly benefited from early materials testing, before the design phase, to reduced organic outgassing and particulate shedding inside the laboratory. While the early biohazard investigations in the SRF may deem the samples safe to release to laboratories throughout the world, sterilization methods need to be defined to safely release Martian samples. One key aspect of enabling simultaneous sample containment and cleanliness is the concept of a Double-Walled Isolator (DWI) that ESA has been pursuing. Further R&D is needed about DWIs, sample handling, and instrumentation interfaces before beginning the SRF site specific design phase. A working isolator/DWI engineering model should be developed and fabricated. Isolator sizes, connections, pass-through antechambers, and configurations and basic interfaces with instruments and tools as well as the use of robotic, mechanical, and/or human manipulation need to be considered. Inert high purity gas supply (one-pass or recirculation) and other isolator utilities must be determined. Special accommodations for large instruments (e.g., XCT,SEM, etc.) need to be developed. Techniques for opening the sample tubes packaged by M2020and extracting the headspace gas while keeping samples pristine will be challenging, requiring additional R&D, and engagement of the M2020 project. In addition, careful micromanipulation for subdivision and packaging of samples will also need to be included. These combined R&D activities are important before commencing the SRF design phase to ensure safe biohazard containment, sample preservation, and science integrity for the samples that would be studied in the SRF and in laboratories around the world.

mars sample return↗

NASA NTRSDate not supplied

Tours of High-containment and Pristine Facilities in Support of Mars Sample Return (MSR) Sample Receiving Facility (SRF) Definition Studies

During 2019 and 2020, the NASA Tiger Team RAMA (acronym of the authors) toured several high-containment biosafety laboratories and pristine space-mission facilities worldwide to better understand their practices, capabilities, and lessons-learned to aid in planning a Sample Receiving Facility (SRF) in support of Mars Sample Return (MSR). The team also included tours of a manufacturer of mobile and modular high-containment facilities as well as manufacturers of isolators and gloveboxes. In addition, the team visited the European Space Agency (ESA)ultraclean and sterile ISO 3 / airborne molecular contamination -9 (AMC-9) isolator line to clean and assemble the most critical hardware for ESA’s ExoMars Mars Lander System, and researchers developing a novel double-walled isolator (DWI) and robotic handling techniques in support of an MSR SRF. The RAMA team visits covered several construction modalities for an MSR SRF: (1) a new traditional fixed facility; (2) use of an existing fixed Biosafety Level 4(BSL-4) facility; (3) a novel modular BSL-4 approach; and (4) a hybrid combination of fixed, modular, and existing facilities. A new fixed facility approach can be tailored to MSR’s needs and is the approach used by all U.S. BSL-4 laboratories constructed to date. However, this approach could be the most expensive modality, take the longest to implement (8-12 years), and have significant programmatic risk of delay. The utilization of an existing BSL-4 facility may be possible depending on the final contamination control and science requirements for the MSR SRF. Due to the internal dimensions of the labs visited and facility structural requirements, it is unlikely that any modification can be made to the facility to meet cleanliness requirements. Furthermore, due to possible construction delays, possible capacity issues, and potential cross contamination vectors from in-house select agents, there may also be significant programmatic risks for sharing an existing facility. Another approach is building a contemporary modular facility. This is a novel approach that has recently been used for a BSL-3/3Ag facilities. The modular elements would be installed in a traditional building or shell structure. A modular facility has many advantages over a traditional fixed facility with lower costs, shorter design/construction/ commissioning schedule, and flexibility for easier retrofits and future expansion. Lastly, a hybrid approach of combining the use of either: (1) a modular facility inside a new fixed facility or (2) a modular and/or fixed BSL-4 annex in conjunction with an existing BSL-4 space should be considered. The advantage of a hybrid approach is that the facility could leverage the strengths of other approaches. Beyond facility construction approaches, the RAMA team investigated technologies and techniques for isolating and handling Martian samples in pristine environments. For example, ESA has been studying and developing a DWI breadboard along with other sample-handling technologies. The research and development investment for clean, remote manipulation and robotics at the start of the facility design phase would be beneficial to the SRF. Additionally, under-standing the lessons learned from Thales Alenia Space during the construction and operation of the most advanced state-of-the-art precision cleaning, sterilization, and assembly glovebox isolators ever developed for spacecraft hardware are also critical for the SRF. The RAMA team lays out a summary of the 18 facilities toured, and includes 43 observations,18 findings, and 22 areas of possible follow-up that the RAMA team and others could pursue to enable further findings. The observations and findings illustrate that constructing an MSR SRF would combine the complexity of both high-containment and pristine facilities, and merging these technologies would be challenging, but achievable.

Mars Sample Return↗

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Mars Sample Return (MSR) Sample Receiving Facility (SRF) Assessment Study (MSAS)

NASA, in partnership with the European Space Agency (ESA), is seeking to return Martian geological and atmospheric samples to Earth for scientific study in the early 2030s. Due to the possibility that the samples could contain extraterrestrial life, Mars Sample Return (MSR) is classified as a Category V: Restricted Earth Return mission by the NASA Planetary Protection Office. As a result of this classification, a MSR Sample Receiving Facility (SRF) must not only provide a pristine environment to ensure samples are protected from terrestrial contamination for scientific investigations, it must also provide high-containment (biosafety level 4 [BSL-4]-equivalence) to isolate the samples from Earth’s biosphere until the samples are deemed safe for release and/or sterilized. The nominal utilization period for a SRF is anticipated to be 2-5 years and is intended to enable curation activities, biohazard assessment, select early science activities, and the rapid release of samples to the scientific community. However, to account for possible delays in schedule or the identification of extant life, this anticipated period of time must be flexible to accommodate schedule extensions and contingency plans. Due to requirements for high-level biological containment and cleanliness, a traditional receiving/curation facility cannot be utilized for MSR. Therefore, beginning in 2022, NASA Johnson Space Center is performing a MSR SRF Assessment Study (MSAS) to investigate the most optimal facility modality for a MSR SRF, as well as start to define programmatic early estimate of costs and schedules before the initial design phase begins. NASA is partnering with industry contractors (architectural and engineering firms with BSL-4 and cleanroom technology experience, as well as other contracted infrastructure and construction specialists) along with selected experts from NASA, ESA, existing U.S. BSL-4 facilities, and other U.S. government agencies, to carry out the assessment study. The MSAS should also aid in the future refinement of the science requirements (e.g., contamination control, equipment accommodations) before site-specific design would commence. As part of the MSAS, NASA is planning to assess an array of possibilities for a MSR SRF. One of the main considerations is the facility modality and whether an existing BSL-4 facility can be utilized (for some or all functions); or, if new constructure would be required, would a traditional fixed facility or a modular facility the best choice. MSAS will also investigate the ability of the modalities to accommodate two different facility capability endmembers: 1) a minimal facility focusing on biohazard assessment and curation tasks with a small footprint, and 2) an enhanced facility with additional capabilities to enable expedited processing and the completion of time-sensitive and (some) sterilization-sensitive science. The assessment is intended to generate information that will inform the selection of facility modalities for high-level conceptual design development. While the assessment study will focus on SRF requirements for accommodating curation, science, and sample safety assessment infrastructure, it will also consider an array of other factors, such as ease of access for international users, decommissioning, repurposing, future sale or lease following MSR’s use of the facility, and uncontained preparatory laboratory spaces. Upon completion of the study, the preferred modality and refined requirements would be utilized for site-specific design but will not be finalized until NASA’s completion of the National Environmental Policy Act (NEPA) process.

A.D. Harrington↗