Search NASA⌕ Search

SEARCH · Search NASA

Results for “planetary protection sterilization”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

103 records · Page 6
All sourcesNASA NTRSDOE OSTI
NASA NTRS2016

EVA-Compatible Microbial Swab Tool

When we send humans to search for life on Mars, we'll need to know what we brought with us versus what may already be there. To ensure our crewed spacecraft meet planetary protection requirements—and to protect our science from human contamination—we'll need to know whether micro-organisms are leaking/venting from our ships and spacesuits. This is easily done by swabbing external vents and suit surfaces for analysis, but requires a specialized tool for the job. Engineers at the National Aeronautics and Space Administration (NASA) recently developed an Extravehicular Activity (EVA)-compatible swab tool that can be used to sample current space suits and life support systems. Data collected now will influence Mars life support and EVA hardware early in the planning process, before design changes become difficult and expensive.NASA’s EVA swab tool pairs a Space Shuttle-era tool handle with a commercially available swab tip mounted into a custom-designed end effector. A glove-compatible release mechanism allows the handle to quickly switch between swab tips, much like a shaving razor handle can snap onto a disposable blade cartridge. Swab tips are stowed inside individual sterile containers, each fitted with a microbial filter that allows the container to equalize atmospheric pressure, but prevents cabin contaminants from rushing into the container when passing from the EVA environment into a pressurized cabin. A bank of containers arrayed inside a tool caddy allows up to six individual samples to be collected during a given spacewalk.NASA plans to use the tool in 2016 to collect samples from various spacesuits during ground testing to determine what (if any) human-borne microbial contamination leaks from the suit under simulated thermal vacuum conditions. Next, the tool will be used on board the International Space Station to assess the types of microbial contaminants found on external environmental control and life support system vents. Data will support advanced EVA and life support system maturation studies, helping to answer questions such as “how close can an EVA-suited crew member approach an area of scientific interest without compromising the science?”

Rucker, Michelle A.↗
NASA NTRSDate not supplied

Overview of the Capture, Containment, and Return System (CCRS)

The Mars Sample Return (MSR) campaign is one of the most ambitious and complex planetary exploration missions currently underway. With the participation of NASA, ESA, and a large number of industry partners, MSR aims to bring Martian soil, rock, and atmospheric samples back to Earth, in order to answer key questions about Mars’ biological evolution. To accomplish this goal the campaign relies on four coordinated missions, each fulfilling a fundamental role to bring the samples to Earth. The Mars Perseverance rover, the first of the four missions, landed safely on Mars on February 18, 2021 and has already acquired candidate samples for Earth return. A selection of the samples of Martian soil and atmosphere that Perseverance has captured during its mission will be recovered, launched into Mars orbit, and transported back to Earth. The Sample Fetch Rover and Mars Ascent System, both parts of the Sample Return Lander project, perform the Mars surface missions to retrieve the collected samples and launch them into Mars orbit. NASA’s Capture, Containment, and Return System (CCRS), hosted on ESA’s Earth Return Orbiter (ERO), brings the samples back to Earth from Mars orbit. These retrieval and return missions are currently in the planning and design stages of development. The NASA-provided CCRS is the payload of the ESA ERO and is the focus of this presentation. ERO will enter Mars orbit and provide communication relay to Earth for the other MSR elements. The Sample Return Lander systems will fetch the sample tubes and integrate them into a protective vessel – the Orbiting Sample (OS) system – which is then launched into low Mars orbit. ERO will perform rendezvous maneuvers, allowing its CCRS payload to capture the OS, contain it, and perform the first automated in-space assembly of a spacecraft, the Earth Entry System (EES), while in Mars orbit. ERO will then begin its journey back to Earth, with CCRS and its assembled EES spacecraft. Three days prior to arrival, CCRS will release the EES on an Earth entry trajectory from a distance beyond the orbit of the Moon. The passive EES spacecraft will then enter Earth’s atmosphere, flying on a ballistic trajectory, followed by a terminal descent (without a parachute) and landing at the Utah Test and Training Range (UTTR). This presentation will show the current design of the CCRS system and its concept of operations. ERO and CCRS will perform several firsts in planetary exploration: (a) orbital rendezvous and capture in Mars orbit, (b) in-space sterilization and containment, (c) on-orbit spacecraft assembly at Mars, and (d) fully-passive entry, descent, and landing sequence for sample return.

Carlie H. Zumwalt↗
NASA NTRS2002

Biomolecular Effects of Cold Plasma Exposure

The effects of cold plasma exposure on Deinococcus radiodurans, plasmid DNA and model proteins were assessed using microbiological, spectrometric and biochemical techniques. Exposure of D. radiodurans, an extremely radiation resistant microbe, to O2 plasma (less than or equal to 25 W, approx. 45 mTorr, 90 min) yielded a approx. 99.999 % sterilization and the sterilization rate was increased approx. 10-fold at 100 W and 500 mTorr. AFM images shows that the exposed cells are significantly deformed and possess 50-70 nm concavities. IR analysis indicates the chemical degradation of lipids, proteins and carotenoids of the cell wall and membrane. Intracellular damage was indicated by major absorbance loss at 1245, 1651 and 1538/cm corresponding to degradation of DNA and proteins, respectively. Biochemical experiments demonstrate that plasmas induce strand scissions and crosslinking of plasmid DNA, and reduction of enzyme activity; the degradation is power dependent with total sample loss occurring in 60 s at 200 W and 500 mTorr. Emission spectroscopy shows that D. radiodurans is volatilized into CO2, CO, N2 and H2O confirming the removal of biological matter from contaminated surfaces. The O2 plasma impacts several cellular components predominantly through chemical degradation by atomic oxygen. A CO2, plasma, however, was not effective at degrading D. radiodurans, revealing the importance of plasma composition, which has implications for planetary protection and the contamination of Mars.

Mogul, Rakesh↗
NASA NTRSDate not supplied

Cleanroom Microbes Survive Drying, Vacuum, and Proton Irradiation

Introduction : The goal of planetary protection at NASA is to mitigate the risk of contaminating sensitive target bodies with biological life. While many cleaning procedures have been put in place to reduce bioburden on spacecraft, microbes are experts at evolving to survive harsh conditions. Specifically, the dry, low-nutrient environment of a cleanroom (commonly used for assembly of spacecraft) can represent an environment where extremophiles can survive. Methods : Scientists at NASA MSFC wished to gather a snapshot of the microbial population within a variety of cleanrooms on site. A study was undertaken to collect air, surface, and floor samples from clean-rooms and isolate unique morphologies. From this study, 95 isolates were collected and saved in a microbial library. About 86% of these were identified at least to a genus level. Following identification, 24 microbes were selected, based on a literature review, as potential extremophiles. These were grown in liquid cultures, diluted to a set optical density, washed with water, and then applied to a sterilized Kapton coupon. Droplets were allowed to dry overnight in a biosafety cabinet. Coupons were then installed in a pelletron and pumped down to high vacuum (~1E-6 Torr). Samples were then subjected 100 keV protons at a fluence of 2x10 15 p+/cm 2 up to 4x10 15 p+/cm 2 . Following exposure, samples were returned to the microbiology lab where they were pro-cessed by submerging in water, vortexing, and then plating either droplets or spread plates. Recovery data collected was qualitative with a ranking or +, minor, or – for growth. Some selected radiotolerant strains were sequenced using the Illumina sequencing platform. The resulting genomes were annotated with the Rapid Annotations using Subsystems Technology (RAST) server and analyzed for conserved and unique stress response relevant genomic signatures to identify clues related to specific tolerances. Results and Discussion : After five rounds of proton radiation, we narrowed our isolates to five, non-spore forming bacteria that demonstrated survival: Arthrobacter koreensis, Paenarthrobacter nitroguajacolicus, Mycetocola manganoxydans , and an Erwinia sp. Furthermore, we exposed these four microbes to 254 nm wavelength light at an intensity of 80 W/m 2 at a distance of ~18 cm for 10 minutes. Only A. koreensis demonstrated survival following UV exposure. Finally, we performed whole genome sequencing on the four strains to look for genetic markers of stress resistance. When we compared the genomes of the four strains, we found that genes coding for GGDEF and EAL domains with PAS/PAC sensors were only found in A. koreensis . These domains, modulated by PAS/PAC sensors, are hypothesized to facilitate survival under drying, desiccation, and proton irradiation. Drying and Desiccation : PAS domains sense hydration changes and modulate GGDEF and EAL domain activity to adjust c-di-GMP levels, enhancing resistance to desiccation. For instance, in Pseudomonas aeruginosa , the PAS domain of RbdA modulates activity under varying hydration conditions, affecting stress responses [1]. Proton Irradiation : Proton irradiation causes oxidative stress, leading to ROS generation. PAS domains detect this stress and modulate GGDEF and EAL domains to manage oxidative stress responses. In Shewanella , EAL domain proteins modulated by PAS sensors help bacteria adapt to extreme conditions [2]. These genes upregulate other stress response genes, protecting membrane function, protein stability, DNA repair, and antioxidant defenses. The modulation of c-di-GMP by PAS domains is crucial for bacterial adaptation to stress conditions, enabling dynamic physio-logical adjustments [3]. Understanding these mechanisms provides insights into bacterial stress responses and strategies for controlling bacterial growth [4]. Conclusions : These findings indicate that clean-rooms harbor extremophile microbes that may be able to survive conditions in deep space. Furthermore, while we identified certain stress-response genes that may be at least partly responsible for the phenotypes observed in this study, there are likely unidentified genes or characteristics about A. koreensis , and other bacteria, that may allow them to survive in harsh environments. Future studies will focus on identifying these unknown genes and characteristics, further elucidating the mechanisms of extremophile survival and potentially informing the development of new biotechnologies for space exploration and other extreme environments.

Chelsi Cassilly↗
NASA NTRS2018

UV Shielding of Bacillus pumilus SAFR-032 Endospores by Martian Regolith Simulants

As exploration of the solar system advances with life detection missions on the horizon, the concern for planetary protection has grown considerably. When attempting to detect extraterrestrial life, the likelihood of false positives from terrestrial contamination must be minimized. The Exposing Microorganisms in the Stratosphere (E-MIST) balloon project aims to evaluate whether resilient terrestrial bacteria can survive stressors in a Mars-like environment. This is accomplished by sending Bacillus pumilus SAFR-032, an endospore-forming bacterial isolate from a spacecraft assembly facility, to the Earth's middle stratosphere (30-38 kilometers), where low temperature and pressure and high radiation and dryness conditions are similar to the surface of Mars. Previous ground and flight tests showed that the vast majority of SAFR-032 spores (99.99 percent) were inactivated by direct sunlight due to ultraviolet (UV) radiation. This observation led us to explore the role of dust shielding in changing microbial survivorship outcomes. To determine the dust particle distributions and density for potentially shielding microbes from UV radiation, samples of a Martian dust simulant were mixed with SAFR-032 spores. The dry heat sterilized simulant used was JSC MARS-1, weathered volcanic ash from Hawaii that displays many chemical and physical properties similar to the Martian soil as characterized by the Viking Lander 1, including reflectance spectrum, chemical composition, mineralogy, grain size, specific gravity, and magnetic properties. First, scanning electron microscopy was undertaken to visualize the aggregation of the spores with dust particles (i.e., shading effects), and samples of varying dust concentrations were subsequently irradiated with UVC light to test survivorship outcomes. After a relationship between dust concentration and spore survivorship was determined, a solar simulator capable of irradiating samples with a fuller UV spectrum (less than 280-400 nanometers) was used to perform a more robust middle stratosphere simulation. Taken together, we will use results from the ground-based irradiation studies to feed into experimental designs for the next E-MIST ultra-long duration polar balloon flight launched by NASA.

Stratosphere↗
NASA NTRS2014

Mars Sample Return Using Commercial Capabilities: Mission Architecture Overview

Mars Sample Return (MSR) is the highest priority science mission for the next decade as recommended by the recent Decadal Survey of Planetary Science. This paper presents an overview of a feasibility study for an MSR mission. The objective of the study was to determine whether emerging commercial capabilities can be used to reduce the number of mission systems and launches required to return the samples, with the goal of reducing mission cost. We report the feasibility of a complete and closed MSR mission design using the following scenario that covers three synodic launch opportunities, beginning with the 2022 opportunity: A Falcon Heavy injects a SpaceX Red Dragon capsule and trunk onto a Trans Mars Injection (TMI) trajectory. The capsule is modified to carry all the hardware needed to return samples collected on Mars including a Mars Ascent Vehicle (MAV), an Earth Return Vehicle (ERV), and hardware to transfer a sample collected in a previously landed rover mission to the ERV. The Red Dragon descends to land on the surface of Mars using Super Sonic Retro Propulsion (SSRP). After previously collected samples are transferred to the ERV, the single-stage MAV launches the ERV from the surface of Mars. The MAV uses a storable liquid bi-propellant propulsion system to deliver the ERV to a Mars phasing orbit. After a brief phasing period, the ERV, which also uses a storable bi-propellant system, performs a Trans Earth Injection (TEI) burn. Upon arrival at Earth, the ERV performs Earth and lunar swing-bys and is placed into a lunar trailing circular orbit - an Earth orbit, at lunar distance. A later mission, using Dragon and launched by a Falcon Heavy, performs a rendezvous with the ERV in the lunar trailing orbit, retrieves the sample container and breaks the chain of contact with Mars by transferring the sample into a sterile and secure container. With the sample contained, the retrieving spacecraft makes a controlled Earth re-entry preventing any unintended release of pristine martian materials into the Earth's biosphere. The analysis methods employed standard and specialized aerospace engineering tools. Mission system elements were analyzed with either direct techniques or by using parametric mass estimating relationships (MERs). The architecture was iterated until overall mission convergence was achieved on at least one path. Subsystems analyzed in this study include support structures, power system, nose fairing, thermal insulation, actuation devices, MAV exhaust venting, and GN&C. Best practice application of loads, mass growth contingencies, and resource margins were used. For Falcon Heavy capabilities and Dragon subsystems we utilized publically available data from SpaceX, published analyses from other sources, as well as our own engineering and aerodynamic estimates. Earth Launch mass is under 11 mt, which is within the estimated capability of a Falcon Heavy, with margin. Total entry masses between 7 and 10 mt were considered with closure occurring between 9 and 10 mt. Propellant mass fractions for each major phase of the EDL - Entry, Terminal Descent, and Hazard Avoidance - have been derived. An assessment of the effect of the entry conditions on the thermal protection system (TPS), currently in use for Dragon missions, shows no significant stressors. A useful payload mass of 2.0 mt is provided and includes mass growth allowances for the MAV, the ERV, and mission unique equipment. We also report options for the MAV and ERV, including propulsion systems, crewed versus robotic retrieval mission, as well as direct Earth entry. International planetary protection policies as well as verifiable means of compliance will have a large impact on any MSR mission design. We identify areas within our architecture where such impacts occur. We also describe preliminary compliance measures that will be the subject of future work. This work shows that emerging commercial capabilities as well as new methodologies can be used to efficiently support an important planetary science objective. The work also has applications for human exploration missions that use propulsive EDL techniques

Red Dragon↗
NASA NTRS2013

Mars Sample Return Using Commercial Capabilities: Mission Architecture Overview

Mars Sample Return (MSR) is the highest priority science mission for the next decade as recommended by the recent Decadal Survey of Planetary Science. This paper presents an overview of a feasibility study for a MSR mission. The objective of the study was to determine whether emerging commercial capabilities can be used to reduce the number of mission systems and launches required to return the samples, with the goal of reducing mission cost. The major element required for the MSR mission are described and include an integration of the emerging commercial capabilities with small spacecraft design techniques; new utilizations of traditional aerospace technologies; and recent technological developments. We report the feasibility of a complete and closed MSR mission design using the following scenario that covers three synodic launch opportunities, beginning with the 2022 opportunity: A Falcon Heavy injects a SpaceX Red Dragon capsule and trunk onto a Trans Mars Injection (TMI) trajectory. The capsule is modified to carry all the hardware needed to return samples collected on Mars including a Mars Ascent Vehicle (MAV); an Earth Return Vehicle (ERV); and hardware to transfer a sample collected in a previously landed rover mission to the ERV. The Red Dragon descends to land on the surface of Mars using Supersonic Retro Propulsion (SRP). After previously collected samples are transferred to the ERV, the single-stage MAV launches the ERV from the surface of Mars to a Mars phasing orbit. The MAV uses a storable liquid, pump fed bi-propellant propulsion system. After a brief phasing period, the ERV, which also uses a storable bi-propellant system, performs a Trans Earth Injection (TEI) burn. Once near Earth the ERV performs Earth and lunar swing-bys and is placed into a Lunar Trailing Orbit (LTO0 - an Earth orbit, at lunar distance. A later mission, using a Dragon and launched by a Falcon Heavy, performs a rendezvous with the ERV in the lunar trailing orbit, retrieves the sample container and breaks the chain of contact with Mars by transferring the sample into a sterile and secure container. With the sample contained, the retrieving spacecraft, makes a controlled Earth re-entry preventing any unintended release of pristine Martian materials into the Earth's biosphere. Other capsule type vehicles and associated launchers may be applicable. The analysis methods employed standard and specialized aerospace engineering tools. Mission system elements were analyzed with either direct techniques or by using parametric mass estimating relationships (MERs). The architecture was iterated until overall mission convergence was achieved on at least one path. Subsystems analyzed in this study include support structures, power system, nose fairing, thermal insulation, actuation devices, MAV exhaust venting, and GN&C. Best practice application of loads, mass growth contingencies, and resource margins were used. For Falcon Heavy capabilities and Dragon subsystems we utilized publically available data from SpaceX; published analyses from other sources; as well as our own engineering and aerodynamic estimates. Earth Launch mass is under 11 mt, which is within the estimated capability of a Falcon Heavy, with margin. Total entry masses between 7 and 10 mt were considered with closure occurring between 9 and 10 mt. Propellant mass fractions for each major phase of the EDL - Entry, Terminal Descent, and Hazard Avoidance - have been derived. An assessment of the entry conditions on the thermal protection system (TPS), currently in use for Dragon missions, has been made. And shows no significant stressors. A useful mass of 2.0 mt is provided and includes mass growth allowances for the MAV, the ERV, and mission unique equipment. We also report on alternate propellant options for the MAV and options for the ERV, including propulsion systems; crewed versus robotic retrieval mission; as well as direct Earth entry. International Planetary Protection Policies as well as verifiable means of compliance will have a large impact on any MSR mission design. We identify areas within our architecture where such impacts occur. This work shows that emerging commercial capabilities can be used to effectively integrated into a mission to achieve an important planetary science objective.

Mars Sample Return↗
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 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 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

The Importance of Contamination Knowledge - Insights into Mars Sample Return

The Astromaterials Acquisition and Curation Office at NASA Johnson Space Center (JSC), in Houston, TX (henceforth Curation Office) manages the curation of all past, present, and future extraterrestrial samples returned by NASA missions and shared collections from international partners, preserving their integrity for future scientific study while providing the samples to the international community in a fair and unbiased way. The Curation Office also curates all reference and witness materials for each mission (e.g., flight and non-flight hardware coupons; lubricants; non-flight, flight-like, and flown witness plates). These reference and witness materials provide the scientific community with the fundamental ability to reconstruct the contamination/alteration history of the sample collection through the course of the mission, with the overall goal of strengthening the scientific conclusions drawn from the study of returned materials. The information gained from characterizing the physical, biological, inorganic, and organic chemical properties of reference and witness materials is defined as the Contamination Knowledge (CK) of the sample collection. Unlike the data collected for Contamination Control (CC) and Planetary Protection (PP), CK is exclusively concerned with preserving reference and witness materials for study by future scientists upon sample return. Although CC and PP data collected for sample integrity and forward contamination purposes can be complementary to CK, they are two separate data sets with distinct objectives. A robust collection of samples for CK is necessary to allow the extraterrestrial material in a returned sample to be distinguished from terrestrial contamination. Traditionally CK is utilized by sample scientists in order to accomplish the mission’s scientific objectives, however this information can also be utilized by the Office of Planetary Protection to help evaluate the presence of any back contamination. Mars 2020, the first phase of a potential multipart Mars Sample Return (MSR) campaign, is expected to contribute to NASA’s Mars Exploration Program Science Goals by filling in knowledge gaps concerning: 1) the existence of past or present life on Mars, 2) the past and present climate of Mars, 3) the geology of Mars, and 4) hazards associated with human exploration of Mars. Although there is debate concerning which samples will best answer these questions, the necessity for proper sample blanks is well-understood. The CC and PP requirements, driven by the restricted Class V mission designation, are the most stringent of any sample return mission in recent history. The extremely low levels of allowable terrestrial contamination on the spacecraft and rover can complicate these analyses given the detection limits of current analytical instrumentation, especially in the case of biological contamination. By collecting and curating unanalyzed samples specifically for CK, future sample scientists will not be relegated to: 1) relying on data collected using possibly obsolete tools and techniques for return sample blanks, or 2) using remnants of extracted and/or cultured samples from ATLO (Assembly, Test, and Launch Operations), which could be incompatible with the desired experimental endpoints or state-of-the-art techniques available at the time of sample return.The addition of biological experimental endpoints to a sample return campaign’s objectives broadens the requisite range in preservation environments (e.g. inert ultra-pure nitrogen gaseous environment at 18 degrees Centigrade versus less than or equal to minus 80 degrees Centigrade) and types of CK samples. As a result, the Curation Office will also curate the following CK samples at less than or equal to minus 80 degrees Centigrade for the Mars 2020 mission: 1) unanalyzed swabs and wipes in sterile containers, 2) all recirculation filters from the clean rooms used for sample and caching subsystem assembly and all filters from the laminar flow benches used to assemble sample intimate hardware, and 3) witness plates collecting airborne contamination within the assembly clean rooms. It has been Curation Office policy since the Apollo missions to preserve as many pristine samples as possible for future scientific research. Although CK is required to be collected for all stages of the MSR campaign, the CK for the Mars 2020 mission is the most critical for understanding contamination in the returned samples given the intimacy between the Martian samples and the Mars 2020 flight hardware. This presentation highlights the importance of CK for sample return missions as well as the traditional and novel types of CK samples required for a successful MSR campaign.

Harrington, A. D.↗
NASA NTRSDate not supplied

Planetary Protection at Marshall Space Flight Center

Introduction: NASA Marshall Space Flight Center (MSFC) is historically known for its role in propulsion. While this is still the mainstay of MSFC’s expertise, many unique capabilities exist at MSFC which pertain to Planetary Protection (PP), including 1) identifying PP threats, and 2) developing novel methods to neutralize those threats. Furthermore, because these capabilities exist among diverse groups at MSFC, this work promotes collaboration both within and outside MSFC to expand and develop PP studies related to a full spectrum of NASA research, design, manufacture, and test interests. This abstract describes the PP research ongoing at MSFC and describes how it contributes to NASA’s overall PP objectives. Microbial Identification in Cleanrooms: One of the greatest threats to successful implementation of PP requirements is recontamination post bioburden reduction. One method to prevent recontamination is to keep the spacecraft in clean environments (i.e. cleanrooms) as much as possible during assembly and integration. However, cleanrooms are not without their own sources of contamination, which is why NASA is interested in monitoring the cleanliness of cleanrooms and characterizing the microbial species present. Such information allows a greater understanding of the resistance of these microbes to cleaning methods, as well as the risk of their contaminating the targeted planetary body of a given mission. MSFC has multiple cleanrooms of various ISO cleanliness levels onsite. We sampled the air and surfaces of three of these rooms, isolated microbes, and then sequenced the 16S rRNA gene or ITS region of the 18S rRNA gene for bacterial and fungal isolates, respectively. This has resulted in a microbial library which currently includes nearly 100 isolates. Microbial Enumeration of Spacecraft Materials: Currently, there are only a couple bioburden reduction methods approved by NASA, and often the harshness of these methods presents additional concerns or risks related to material properties. The goal of this research is to assess the microbial content of solid rocket motor (SRM) materials potentially used for lander missions. This work aims to more accurately define the risk of planetary contamination by providing empirical data associated with commonly used SRM raw materials. In this study, we pulverized nonmetallic SRM materials using a cryogenic grinder, then analyzed the resulting substrate for microbial colony forming units (CFU). We found that many SRM nonmetallic materials do not harbor detectable bioburden, though a range existed depending on the material. The results from this work provide quantitative data to potentially reduce concerns of contamination, while also providing a foundation for follow up studies into additional sterilization methods and molecular identification of contaminating microbes. Space Environmental Effects on Microbial Survival: One potential area of microbial reduction is the space environment. Understanding the survivability of hardy microbes in space-like conditions is a crucial first step in answering how space may reduce bioburden and if it can be relied upon for adherence to PP requirements. This work studied the effects of ultraviolet (UV) and ionizing radiation on survival of Bacillus atrophaeus spores. Microbes were dried on relevant polymeric materials then exposed to space environmental stressors. Coupons were submerged in water, diluted, and plated to determine survival compared with controls. We found that both UV and ionizing radiation were capable of reducing viability by nearly 99%, but there were still survivors, some with changed morphology indicating resistance mechanisms within certain cells. Manufacturing credit: Finally, given the above-mentioned limitations of the NASA-approved bioburden reduction methods, there is interest in understanding if manufacturing processes may provide enough bioburden reduction without additional PP-specific bakeouts. For instance, some material additives may be antimicrobial. Given this, we investigated the effects of several commonly used rubber additives on the growth of B. atrophaeus spores. We found that some of the materials inhibited growth of the spores, possibly supporting the use of these additives on missions with PP constraints. Future work into manufacturing credit for bioburden reduction includes inoculation of green insulation with B. atrophaeus spores, followed by a typical cure. Thermal profiles will be verified for appropriate temperature and durations to meet PP requirements, and cured samples will be analyzed using a cryogenic grinder to determine survivability of spores.

Chelsi D. Cassilly↗
NASA NTRSDate not supplied

EVA Swab Kit: Tools and Techniques for Collecting Aseptic Samples from Crewed Space Missions

Introduction: 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 spacecraft meet planetary protection requirements—and to protect our science from human contamination—we'll need to assess and verify whether micro-organisms may be leaking/venting from our spacesuits. This requires collecting samples under Extravehicular Activity (EVA) conditions. Detailed, systematic research on forward contamination from robotic spacecraft has been steadily progressing since the Viking missions, but systematic studies of contamination from space suits has not been conducted in many years. The modern EMU (Extravehicular Mobility Unit) suit used by NASA is designed to leak at rates as high as 100 cc/min. Before humans land on Mars there is a critical need to understand the types and quantities of microbes that could be introduced via space suits. The Human Forward Contamination Assessment team at NASA’s Johnson Space Center (JSC) has developed a prototype EVA swab tool [1,2,3,4] designed for use in space to sample cleaned and uncleaned space suits to determine the present day microbial load and eventually the rate of leakage. The ability to assess microbial leakage early in advanced space suit and life support system design cycles will help avoid costly hardware redesign later. Test Objectives: The primary objective of EMU testing was to characterize the type of micro-organisms typically found on or near selected suit pressure joints under suit differential pressure conditions. Most human-borne microbes can fit through a 0.5 to 1.0 µm gap. Knowing which joints are more likely to leak will inform hardware design decisions. Knowing which types of micro-organisms 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. That data, in turn, will inform exploration mission operations and hardware design. The secondary objective of testing was to evaluate the interface between a fully suited test subject and the EVA swab tool at vacuum. Bulky EVA suits can restrict movement and limit visibility through the helmet visor. Fully suited testing is important for identifying tool design issues prior to flight. At exploration destinations, such as Mars, suited crew may be required to periodically sample their suits as part of an environmental monitoring protocol. Suit Microbial Sampling Results: This report details results of microbial swabs collected from current flight suit configurations worn by crew members assigned to upcoming ISS expedition missions as well as swabs collected from prototype suits intended for use on the Orion spacecraft. These tests were intended to characterize the types of contaminants found on flight suits under current, typical handling conditions. No attempt was made to change suit handling procedures, provide additional sterilization, or to limit typical potential contaminant sources. Using culture based techniques, we cultivated 235 CFU (colony forming units) comprised of 26 bacterial species and one fungal species on the outside of the suits. The fungal species and 14 of the bacterial species were unique to the suit surfaces and were not detected in any of the background samples collected within the chambers. We sequenced 755,434 ribosomal fragments on all of the suit surfaces from swab samples. 557,016 of these sequences represent DNA that survived at least 4 hours at vacuum. These sequences formed 2,464 OTU's (Operational Taxonomic Units, 97% similarity) showing low diversity in the samples. The most abundant sequences that survived vacuum belong to the genera Staphyloccocus, Ralstona, Bacillus and Rhodobacter all of which are common to the human microbiome. [5] See Danko et al., (2021) for more complete details of these first analyses. Further analysis of EVA suit materials with respect to the efficacy of various cleaning protocols and engineered containment solutions is planned to inform suit design for NASA’s Artemis Moon to Mars program crew testing. Swab Tool Function Results: The kit was demonstrated for fit and function in suited subject vacuum tests to determine how well the tool worked as an aseptic microbial sampling device as well as to identify any design elements that could be upgraded for EVA task specific improvement. It was found that sample acquisition efficacy could be enhanced by redesign of the sample canister to end-effector interface. Several modifications of the sample caddy assemblies to optimize EVA safety and functionality were also identified. Consequently, fabrication of the redesigned sample canister to end-effector assembly interfaces and and the sample caddy assemblies are required. Fabrication of sixteen flight sample canister assemblies (8 per each of two EVA Swab Kits) and two sample caddy assemblies are in process to be followed by hardware testing and certification to produce two flight-certified EVA Swab Kits for transport to ISS no earlier than summer of 2022. Sampling Strategy: The International Space Station is an ideal testbed for systematic studies of contamination from crewed vehicles since it has been continuously occupied for 20 years and exposed to non-terrestrial conditions. We will sample the exterior of the ISS during EVA using a purpose-built swab tool capable of maintaining sterility while undergoing temperature changes from -151 to +121°C under hard vacuum. Prior to each EVA, the project team will work with ISS mission managers to identify precise sampling locations, which will vary by EVA based on the translation paths and worksites scheduled for that particular EVA. Ideally, translation path handrails and areas near ECLSS (Environmental Control and Life Support System) external vent openings on a spacecraft would be assessed. There are currently more than a dozen ECLSS external vents on the ISS. Some are connected to systems that vent waste products, while others are intended to equalize cabin pressure. As EVA opportunity allows, microbial samples from any of these external vents would provide a valuable data point, though some will be more useful than others. Four criteria have been identified to help prioritize sampling sites near vents: • EVA Accessibility: To minimize cost, it is desired to piggy-back onto a planned EVA. Therefore, the sampling location must be readily accessible by an EVA crew • Type of Vented Products: Vent products that have been in direct contact with crew, such as cabin air, are more likely to contain microorganisms than vent products associated with isolated systems, such as experiment module combustion products. • Mass of Vented Products: Higher-flow vents are more likely to contain detectible levels of microbial contaminants than lower-flow vents. • Local Environment: Sample locations with relatively benign local conditions, such as warm surfaces shielded from direct ultraviolet (UV) radiation exposure, may be more likely to support microbial growth than locations with harsher local environmental conditions. Because EVA accessibility is the most important criteria, the proposal team worked with an astronaut and flight controllers using the Dynamic Onboard Ubiquitous Graphics (DOUG) tool. The DOUG virtual environment allows an operator to “fly” around the current ISS vehicle configuration to assess EVA translation paths, attach points, and keep-out zones. While analysis on station or rapid return to Earth would be preferable, samples collected from the exterior of the ISS have already been exposed to temperature variations between -157 and +121 °C as well as hard vacuum. Therefore, they should be fairly stable and robust. We hypothesize that samples collected from the ISS exterior could be stored for up to 6 months at -80°C without degradation. Sample canisters will be returned to Earth while frozen at -80°C for analysis, and sterilized canisters can be re-flown back to ISS to support additional sampling opportunities Relevance to NASA Exploration Objectives: These data will allow us to identify new or improved methods, technologies, and procedures for spacecraft sterilization and leakage mitigation to minimize the amount of contamination introduced to the environment by human explorers. This work is funded by NASA research grant: NNH18ZDA001N-PPR References: [1] Bell, M.S. et al. (2015) LPS XLVI, Abst. #1832 [2] Rucker et al. (2018) 42nd COSPAR (PPP.3) [3] Bell, M.S. et al. (2019) Mars Extant Life Conference, Abst. #5096.[4] Bell, M.S. et al., (2020) 43rd COSPAR (BO.2).[5] Danko D, et.al.,(2021)Front.Microbiol.12:608478.

Mary Suzanne Bell↗