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NASA NTRSDate not supplied

Assessing Risk Due to Small Sample Size in Probability of Detection Analysis Using Tolerance Intervals

Small sample size (e.g.6-30) poses risk in results of probability of detection (POD) analysis using tolerance intervals. This method is also called as the limited sample or LS POD. The analysis is performed either during NDE procedure qualification or for assessment of reliability of an NDE procedure. The risk is primarily due to sampling error. Smaller samples are not likely to be random to the population or representative of the population. The small samples are likely to be biased. Biased samples have smaller standard deviation compared to the population. POD analysis with small biased sample can lead to overestimation of POD. Many sampling schemes are available in statistics to mitigate sampling risk. Primary objective of POD analysis is to determine a decision threshold from signal response measurements of a sample such that it is less than or equal to population decision threshold for 90% POD. Sampling error implies that this NDE reliability condition is violated. One of sampling types is called a representative sample. Representative samples reduce variance in POD estimates but also reduce magnitude of the error. Sampling sensitivity analysis for some sampling types is performed here using repetitive random sampling or Monte Carlo method. Six sampling types are considered for comparison. Some of the sampling types are similar to drawing a representative sample. LS POD model assumes random sampling. Therefore, random sampling is used as a basis for comparison with each sampling type. The sampling types used in the analysis are, A. Nominal and worst-case sampling, B. Worst-case sampling, C. Nominal case sampling, D. Random sampling, E. Random target, and sub-target sampling. F. Nominal target and sub-target sampling. Results of Monte Carlo simulation indicate that type F sampling can mitigate sampling risk and is also more practical to implement. Type A sampling may also mitigate the sampling risk, but it may be less practical to implement.

Ajay M Koshti↗
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

The Effect of Temperature on the Preservation of Volatile-Rich Lunar Samples

Introduction. The Moon’s south pole is a high-priority target for human exploration and scientific study. This interest is, in part, due to the presence of Permanently Shadowed Regions (PSRs), which could contain high concentrations of unique volatiles at cryogenic temperatures [1]. Returned samples from PSRs may include a unique combination of rocks, regolith, and volatile species, providing unprecedented insights into the history of the Solar System and the potential for resource utilization on the Moon. However, because PSR samples are cryogenic up-on collection, lunar polar sample return will eventually require cold stowage for the journey from the Moon to Earth. Without cold stowage, PSR sample return will likely result in phase changes and chemical reactions within the volatile component of the sample, which would negatively impact the resulting scientific studies of those samples. This abstract summarizes the initial results from an ongoing characterization of analog PSR samples at a range of temperatures, with the goal of defining the temperatures needed for a flight cold stowage freezer. Background. Based on remote sensing observations of the Moon [2], south polar PSRs range in temperature from ~120K for small and/or shallow PSRs to ~20K at the most extreme locations in large, deep PSRs. A range of volatiles have been hypothesized to exist at the surface or subsurface of the lunar poles [3-5 and others]. This hypothesis was verified when the LCROSS mission impacted the <50-K PSR in the crater Cabeus, detecting a range of volatiles from water to low condensation temperature species such as H2S and methane [6]. Species such as H2S and ammonia (also detected by LCROSS) are also highly reactive, and increase the likelihood of chemical reactions at elevated (non-cryogenic) temperatures. At the Johnson Space Center’s Planetary Exploration and Astromaterials Research Laboratory (JSC-PEARL), we have developed a volatile-bearing lunar simulant that incorporates several of the species detected by LCROSS [Table 1] mixed cryogenically with the USGS Lunar Highlands Type (LHT) regolith simulant. The new volatile-regolith simulant will be used to assess the degree of sample alteration at room temperature, -20°C, -80°C, and -196°C (liquid nitro-gen), over a two-week period. Room temperature samples represent those likely to be returned during initial missions without cold stowage, -20°C provides an analog to Apollo cold curated samples, -80°C is the temperature of multiple flight payload freezers (e.g., MELFI), and -196°C is analogous to lunar PSRs. Two weeks is an approximation of the time between sample collection and Earth return for initial Artemis missions. Over this period of time, sample head-space gases will be analyzed using a Universal Gas Analyzer (UGA, a type of mass spectrometer) coupled with a Baratron pressure sensor. After testing, the regolith component of the simulant will be purged of volatiles and preserved for future electron beam and/or FTIR analysis. Experimental Procedure. Volatile-regolith simulants will be produced as an initial homogenous batch; this batch will then be distributed into aliquots (gas chromatography/GC vials or cryo vials), ensuring that each sample has the same starting composition and conditions [Fig. 1]. In addition to the “full” simulant shown in Table 1, less complex simulant compositions will be used as baseline and control samples [Table 2]. Aliquots will be produced in triplicate for each simulant composition, storage temperature, and date of sampling. Headspace gases in all Day 0 samples will be analyzed by the UGA immediately. Cold storage samples for future analytical days will be placed in freezers appropriate to their target temperatures (-20°C, -80°C, -196°C). For ambient-temperature samples, regolith and regolith-water samples will be stored in a fume hood, while the full simulant will be stored in a sealed Parr vessel for safety; no other simulants (RWCM/ RWCM+) will be stored at ambient temperature for this test. Samples will be analyzed by UGA in this manner on each Analysis Day outlined in Table 2. Analytical Data. The UGA measures the partial pressures in a single sample aliquot over a set mass range of 0-105 atomic mass units (AMU) [Figure 2]; this set range was selected to slightly exceed the mass of the highest-mass expected reaction product (H2SO4). The Baratron complements the UGA by measuring the total pressure in the headspace of a sample vial. Coupled together, the quantitative abundances of gases will be monitored throughout the test. UGA analyses of the triplicate samples for each storage temperature, day, and simulant composition will be averaged, and standard deviations for each will be calculated. The compositions of starting species (shown in Table 1) will be characterized as a function of time, and the presence of any new compounds (reaction products) will be monitored as well. Total pressures will be recorded for each sample analysis, and any samples that show signs of leakage (e.g., a significant reduction in pressure or simulant volatiles) will be discarded. Anticipated Results. Testing is planned to begin in January 2022. The resulting data will allow compositional and phase changes in the volatile component of the simulants to be determined. Both the reduction in initial compounds and the addition of reaction products are expected to be observed. In addition, the relative efficacy of the different temperatures at pre-serving the initial composition of the simulants will be quantified. Finally, the regolith component of each sample will be argon-purged and stored in a controlled environment for future laboratory analysis. Compositional and morphological changes in the regolith are expected for samples above 0°C. This test will be repeated three times over the course of 2022. Understanding the effect of temperature on both the volatile and regolith components of analog lunar materials will allow requirements for a cold stowage freezer to be developed. The implementation of cold stowage for lunar polar missions will maximize the preservation of returned samples, enabling ground-breaking lunar and Solar System volatiles science for decades to come.

J L Mitchell↗
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↗
NASA NTRSDate not supplied

The Collection, Usage, and Preliminary Examination of the Apollo Sample Suite: Lessons for Artemis

Apollo Sample Collection and Usage: From 1969 to 1972 there were six Apollo missions to the surface of the Moon during which the astronauts collected 382 kg of rock and regolith (~2200 samples). The samples collected fall into these general categories: rocks (~66% by mass), rake samples (~4%), bulk regolith (~24%), and specialty regolith (deep drill cores, drive tubes, sealed bulk regolith) samples (~6%). In each category there are a variety of different subtypes available for study, e.g., among the bulk regolith samples there are also skim, trench, and (partially) shaded regolith samples each sampling unique types or depths of regolith. This variety of subsamples has enabled a multitude of different studies over the past 55 years (>3400 individual requests). We are still averaging ~50 unique requests and have allocated >500 individual Apollo samples annually for the past 10 years (2020 excepted). Looking at the 4,675 non-ANGSA (Apollo Next Generation Sample Analysis) samples allocated over the past 10 years, the proportions of allocated samples do not precisely align with the abundance (by mass) of those samples withing the collection: Rock (69.4 %); Rake (10.8 %); Bulk Regolith (15.5 %); Drive Tube (3.3 %); Core/Specialty (1.0 %). Apollo Preliminary Examination (PE): The PE process differs significantly for the various sample types enumerated above; we focus on regolith and rock samples here. During the Apollo mission era, the PE process evolved over the course of the missions; below is what was done for the Apollo 17 mission. For regolith samples, the PE process was: (1) documented bags containing regolith are opened, photographed, and described; (2) large rocks are removed and treated separately; (3) 25% to 33% of the bulk soil is scooped out, weighed, and stored in reserve; (4) the remaining sample is sieved to produce the size fractions <1, 1- 2, 2-4, and 4-10 mm, all of which are weighed. For rock samples, the process is: (1) removing rocks from the container(s) it came back from the Moon in; (2) rematching any materials that spalled off the rock to their original location; (3) numbering, weighing, and basic photographic documentation; (4) dusting with a gentle N2 gas jet; (5) Orthogonal photography; (6) detailed description of the textures and features of the rock; (7) rock modelling and measurement; (8) stereophotography; (9) determination of the orientation of the rock on the lunar surface. Drive tubes and deep drill cores were not characterized during PE beyond an initial weight and a sketch of the interior tube materials derived from 2D medical X-ray images. Catalogs: The ongoing utility of the Apollo samples is enabled by the robust cataloguing process for the samples [3-5], which allows the scientific community to accurately request samples uniquely suited to their proposed studies. A common misconception, however, is the amount of detail that goes into the initial catalog (e.g., [6]) for a collection from the preliminary examination (PE) period, versus what goes into the catalogs that come later in the life cycle of the samples from that mission (e.g., [7]). The only required data for a PE catalog is a weight, a basic photograph, and a description of the nature of the sample. Artemis PE: Over the past few years, the ongoing ANGSA project studied previously unopened Apollo 17 double drive tube samples 73001/2 [1], and a PE of the drive tubes was done. The PE took the existing core dissection process (developed during PE of Apollo cores in the 1970s, 1980s), and modernized it [7]. The main lesson from the ANGSA PE relative to future missions was that the physical work done during PE of lunar samples has not changed much over the past 5 decades. The use of “modern” technology during PE (e.g., XCT; multispectral analyses) resulted in an enhanced initial understanding of the 73001 and 73002 drive tubes, but greatly increased the time required. The lessons learned from recent astromaterial PEs (e.g., ANGSA and OREx) are important to consider when planning for Artemis, but the unique nature of the Artemis Campaign means many lessons learned from these mission will not be applicable. Given the time constraints (6 months) and likely number of samples that will be returned by Artemis (>200), the Artemis PE catalog will necessarily look much more like [4] than [6].

J Gross↗
NASA NTRSDate not supplied

OSIRIS-REx Curation: The First Year with Bennu Samples on Earth

The OSIRIS-REx (Origins, Spectral Interpretation, Resource Identification, and Security – Regolith Explorer) sample return capsule landed on the Utah Test and Training Range (UTTR) on September 24, 2023. Preparations for the curation of the sample from asteroid Bennu inside of this capsule spanned more than a decade. On the morning of landing, the sample return capsule was brought to a temporary cleanroom at UTTR for initial de-integration and connection of the sample canister to a nitrogen gas purge to protect the sample from modification by the terrestrial atmosphere. In the week following landing, the sample canister was flown to Houston while on nitrogen purge, transferred to a nitrogen atmosphere glovebox at NASA Johnson Space Center in which the canister lid was removed revealing small amounts of sample on the inside of the canister lid and avionics deck, some of this sample was allocated rapidly to the mission sample analysis team as “quick look” samples. Next, the Touch-and-Go sample acquisition mechanism (TAGSAM) head was disassembled inside of a specially designed glovebox. A challenge arose during one of the last stages of disassembly when two fasteners resisted being loosened. The curation team was able to remove 70.3 grams of asteroid sample from the TAGSAM head with those fasteners in-place and began initial sample processing which supported long-term curation of hermetically sealed and frozen samples, transfer of some samples to remote storage, and allocation of aggregate samples to the mission sample analysis team. Meanwhile, curation engineers developed a tool to remove the stubborn fasteners without contaminating the remaining sample, enabling the remaining bulk sample to be transferred to long-term storage trays in January 2024. With this second part of the bulk sample, the returned bulk asteroid sample reached 121.6 grams. During disassembly and prior to sealing the bulk sample trays into long-term storage containers, archival photographs were collected. Select particles are gradually being XCT scanned while inside of containers that hold a nitrogen atmosphere. The initial OSIRIS-REx sample catalog was released in spring 2024 with an update in fall 2024. The curation team will be busy working to allocate samples to scientists around the world for the foreseeable future.

Curation↗
NASA NTRSDate not supplied

The Genesis Non-flight Implant Sample Archive

Introduction: For the past 20 years the Genesis science community has been analyzing solar wind captured in sample collectors now curated at Johnson Space Center (JSC). Non-flown flight like collector materials (non-flight) considered reference material was always allocated by the JSC Genesis team for testing experimental procedures. However, to expedite Genesis science, distribution of non-flown reference materials were also provided by, D. S. Burnett (Genesis PI, Caltech) and A. J. G. Jurewicz (ASU) who were funded to aid all Genesis PIs by providing engineering test materials archived at Caltech pre-flight as well as ion implants into these materials [1]. Implanted materials (non-flight reference, flight-like semiconductor materials, and NIST SRM glass) received from the Caltech collection during a pilot project are being used to develop an accession procedure for the purpose of making these materials available to researchers in the future. These implant samples are important because they can be utilized as measurement standards [2]. This is a work in progress and this work reports progress in documentation and imaging. The pilot program received 63 implanted reference materials with known implanted ions. Genesis Non-Flight Database: The existing non-flight material database was modified to allow for the description of implant sessions and to allow correlation of the implant information to individual specimens. An implant session has three main components: a known ion, a known dose (ion/cm2), and a known energy (keV). Procedure: Laboratory Procedure. Upon receiving non-flight analytical standards (independently calibrated implants), or implant samples (nominally calibrated implants), they will be imaged in their current containers to capture any information written before being brought into the Genesis Sample Laboratory. Documentation is provided by the donor and used for correlating the data between the implant sessions and implant samples. This information is not verified by Genesis Curation. The microscope used to image samples in the laboratory has an automated stage with a stainless-steel plate to hold samples and a camera that utilizes the software Surveyor to take photomosaic images of the implant samples. The camera attached to the microscope is a QICAM High Performance IEEE 1394 FireWire Digital CCD Camera. This setup, shown in Fig. 1, allows for photomosaics of small-scale samples with a 10x objective or 25x objective. Examples of images taken with this microscope setup are shown in Fig.2 and Fig. 3. After imaging, samples get stored in a clean fluoroware container or polypropylene vial depending on sample size for long term storage. Database Procedure. The Genesis Non-Flight Database currently allows for the addition of implant sessions. The required fields for adding an implant session are the implant session ion, the implant session dose, and the implant session energy. There are also fields for the date and vendor of the sessions as well as a field for if the session was calibrated or not. Once an implant session has been inserted into the database, additional documentation from the session can be attached. The database will automatically assign an Implant Session Number. After inserting an implant session, the implanted samples can be inserted. The fields required for an implant sample are the material of the sample and the size of the sample. The Generic name for the implant sample with be automatically generated by the database once it has been inserted. Implant sample numbers begin with 3X to distinguish from flown Genesis sample numbers. There are also fields for processors to add the location (room, cabinet, tray) and container type (fluoroware, polypropylene vial). Once an implant session and an associated sample(s) have been inserted into the database the sample can be then tied to all associated implant sessions. An implant sample can be tied to multiple implant sessions. Future work: For curation, future work consists of creating an online catalogue for implant samples as well as updating the database with more samples as we receive them. For the scientific community these implant samples have many potential uses. For Genesis research, they are directly applicable for testing and, in some cases, as analytical standards. For non-Genesis research, implants (especially calibrated implants) could be used for calibrating other, unique planetary materials [1].

C D Calva↗
NASA NTRSDate not supplied

Initial analysis of “stone” size Ryugu samples: current status

As a part of the initial analysis of the Ryugu samples, we perform a variety of analyses of millimeter-sized "stones". Our goals are to elucidate the entire formation process of C-type asteroid Ryugu from the viewpoint of petrology and mineralogy ande obtain necessary information by sample analysis, and then simulate the formation of Ryugu based on the evidencef obtained rom sample analysis. Eighteen stones (8 from Room A and 10 from Room C) were received from the ISAS curation facility on June 1, 2021, and brought into a fully nitrogen-displaced glove box at Tohoku University. At the same time, we also received the powder samples from Room A and Room C. To completely block the atmosphere from leaking into the container, all samples were put in the sample transport containers prepared by ISAS, transferred from the main chamber to the glovebox at ISAS, and then all containers were completely sealed in plastic bags with moisture and oxygen absorbers. No moisture or oxygen was detected when the bags were opened in the glove box at Tohoku University, so it was confirmed that there was no exposure to the atmosphere during transport. To date, a number of analyses have been carried out successfully and almost on schedule. The analysis started with the measurement of reflectance spectra, which are sensitive to atmospheric oxidation, hydroxylation, and adsorbed water. The visible, near-infrared, and mid-infrared reflectance spectra were measured while the samples were kept airtight. The spectra of powder samples and stone samples (as aggregates and as single stone) were successfully obtained. A major feature of the stone team's analysis is the use of synchrotron radiation facilities around the world. Since this analysis is non-destructive, stone samples whose reflectance spectra were measured were sent to KEK, SPring-8, ESRF (France), SOLEIL (France), DESY (Germany), and APS (USA). Using these synchrotron radiation facilities, high spatial resolution and sensitivity XRD, STXM, XANES, CT [1], IR-CT, FT-IR [2, 3], XRF, and Mössbauer analyses were performed. Most of the analyses were carried out under air-tight conditions on the stone samples and the particulates separated from the stone samples. These analyses allowed us to determine the three-dimensional distribution of minerals and elements, redox state, density and porosity of the stone samples. Furthermore, as a characteristic analysis of the stone team, light elemental analysis using negative Muon was performed at the MLF facility of J-PARC with an exceptionally long allocation of machine time. This is the only non-destructive method to measure the concentration of light elements in the whole (not the surface) of stone samples. Because the characteristic X-rays produced by muon irradiation are much higher in energy than the fluorescent X-rays produced by X-ray irradiation, there is little effect of self-absorption by the sample, and therefore, the concentration of light elements such as carbon, oxygen, and Na in the entire "stone" sample can be determined. Some stone samples are currently being measured for heat and strength physical properties in order to understand the physical properties of asteroid Ryugu. The data obtained from these measurements are useful for interpreting the remote-sensing data data taken from the surface layer of the asteroid Ryugu [4-7]. It is also important for understanding the behavior of the Ryugu material during impact events. The surfaces of many stone samples were observed by electron microscopy and other techniques, especially on natural “flat” surfaces formed on 5 stomes. As a result, characteristric mineral aggregates formed by the reactions with water and characteiristic impact features were observed on some of the samples. Based on the observations of surfaces and the synchrotron measurements of the whole stones, important objects such as characteristic structures and specific crystal aggregates for understanding the formation history of the asteroid were identified, and these parts were separated from the stone samples using an Xe beam (pFIB) and analyzed by various methods including transmission electron microscopy and synchrotron radiation analysis. Many stone samples, from which important objects have been separated, are embedded in epoxy resin and cut to produce many polished sections. Electron microscopy and spectroscopic measurements of the polished surfaces are being carried out to reveal the detailed mineralogical properties and elemental distribution inside the stone samples. In the fall, machine time for synchrotron radiation will begin, and we plan to analyze single crystals and characteristic objects separated from the stone samples.

Tomoki Nakamura↗
NASA NTRS2002

Technology Development and Advanced Planning for Curation of Returned Mars Samples

NASA Johnson Space Center (JSC) curates extraterrestrial samples, providing the international science community with lunar rock and soil returned by the Apollo astronauts, meteorites collected in Antarctica, cosmic dust collected in the stratosphere, and hardware exposed to the space environment. Curation comprises initial characterization of new samples, preparation and allocation of samples for research, and clean, secure long-term storage. The foundations of this effort are the specialized cleanrooms (class 10 to 10,000) for each of the four types of materials, the supporting facilities, and the people, many of whom have been doing detailed work in clean environments for decades. JSC is also preparing to curate the next generation of extraterrestrial samples. These include samples collected from the solar wind, a comet, and an asteroid. Early planning and R\&D are underway to support post-mission sample handling and curation of samples returned from Mars. One of the strong scientific reasons for returning samples from Mars is to search for evidence of current or past life in the samples. Because of the remote possibility that the samples may contain life forms that are hazardous to the terrestrial biosphere, the National Research Council has recommended that all samples returned from Mars be kept under strict biological containment until tests show that they can safely be released to other laboratories. It is possible that Mars samples may contain only scarce or subtle traces of life or prebiotic chemistry that could readily be overwhelmed by terrestrial contamination . Thus, the facilities used to contain, process, and analyze samples from Mars must have a combination of high-level biocontainment and organic / inorganic chemical cleanliness that is unprecedented. JSC has been conducting feasibility studies and developing designs for a sample receiving facility that would offer biocontainment at least the equivalent of current maximum containment BSL-4 (BioSafety Level 4) laboratories, while simultaneously maintaining cleanliness levels equaling those of state-of-the-art cleanrooms. Unique requirements for the processing of Mars samples have inspired a program to develop handling techniques that are much more precise and reliable than the approach (currently used for lunar samples) of employing gloved human hands in nitrogen-filled gloveboxes. Individual samples from Mars are expected to be much smaller than lunar samples, the total mass of samples returned by each mission being 0.5- 1 kg, compared with many tens of kg of lunar samples returned by each of the six Apollo missions. Smaller samples require much more of the processing to be done under microscopic observation. In addition, the requirements for cleanliness and high-level containment would be difficult to satisfy while using traditional gloveboxes. JSC has constructed a laboratory to test concepts and technologies important to future sample curation. The Advanced Curation Laboratory includes a new-generation glovebox equipped with a robotic arm to evaluate the usability of robotic and teleoperated systems to perform curatorial tasks. The laboratory also contains equipment for precision cleaning and the measurement of trace organic contamination.

Lindstrom, David J.↗
NASA NTRSDate not supplied

The Comet Astrobiology Exploration Sample Return (CAESAR) Mission

The Comet Astrobiology Exploration Sample Return (CAESAR) mission will acquire and return to Earth for laboratory analysis a minimum of 80 grams of surface material from the nucleus of comet 67P/Chur-yumov-Gerasimenko (67P). CAESAR will characterize the surface region sampled, preserve the collected sample in a pristine state, and return evolved volatiles by capturing them in a separate gas reservoir. NASA Goddard Space Flight Center provides project management, systems engineering, safety and mission assurance, contamination control, mission operations, and many other important functions. Northrop Grumman Space Systems will build the spacecraft, based on Dawn mission heritage, which like CAESAR, uses solar electric propulsion. CAESAR was selected by for Phase A study in the New Frontiers 4 Competition and will be proposed to New Frontiers 5.Collection of a sample from the surface of comet 67P is facilitated by a set of cameras that together provide images to support sample site selection, perform optical navigation, and document the sample before, during, and after col-lection. The sample is collected at the end of an arm during a 5-second touch-and-go (TAG) maneuver with the Sample Acquisition System (SAS)designed by Honeybee Robotics for the surface properties of comet 67P observed by the Rosetta mission. After sample collection, and while the sample is still cold (< -80°C), the TAG Arm inserts the sample container into the Sample Containment System (SCS) mounted inside the Sample Return Capsule (SRC). The SCS is sealed, preventing the sample from escaping into space. The sample is slowly warmed inside the SCS to enable sublimation of volatiles, which are collected in the Gas Containment System (GCS), a passively cooled gas reservoir. Separating the volatiles from the solid sample protects the solid sample from alteration. Once all sublimated H2O is transferred to the GCS, the GCS is sealed to capture the volatile sit contains, and the SCS is vented to space to maintain the solid sample under vacuum. The SCS vent is closed before Earth entry to prevent atmospheric contamination. Detailed laboratory analyses of the sample from 67P will trace the history of volatile reservoirs, delineate the chemical pathways that led from simple interstellar species to complex molecules, constrain the evolution of the comet, and evaluate the role of comets in delivering water and prebiotic organics to the early Earth. CAESAR will achieve these goals by carrying out coordinated sample analyses that will link macroscopic properties of the comet with microscale mineralogy, chemistry, and isotopic studies of volatiles and solids. Most of the sample (≥75%) will be set aside for analyses by generations of scientists using continually advancing tools and methods, yielding an enduring scientific treasure that only sample return can provide. This presentation will review development conducted during NF4 Phase A and discuss the NF5 mission concept.

A G Hayes↗
NASA NTRSDate not supplied

Microbial Monitoring of New Cleanrooms Used to Curate Astrobiologically Relevant Asteroid Samples from Bennu and Ryugu

Introduction: NASA has constructed two new cleanrooms to house materials from the OSRIS-REx and Hayabusa2 missions to the asteroids Ryugu (162173) and Bennu (101955), respectively. In accordance with standard astromaterials curation practices, these cleanrooms will be monitored for particulate contamination and maintained to ISO 5 equivalent standards1. Since the samples in these collections are expected to contain prebiotic organic compounds that may help explain the origin of life on Earth, these labs will also be monitored for organic and biological contamination2. Samples from Ryugu arrived on Earth in December, 2020. After basic characterization in Japan, NASA received a subset of these samples at the astromaterials curation facility in Houston in December of 2021. OSIRIS-REx is expected to return samples in September, 2023. Here we present preliminary microbial monitoring results from monthly monitoring of these new labs and the connected microtomy and staging areas that support them, as they are being commissioned. We also compare these results to baseline values for other astromaterials curation labs. We will also briefly describe additional cleaning efforts employed to reduce the bioburden in these new cleanrooms. Methods: Microbial samples were collected from surfaces using a dry macrofoam swab (Puritan Brand 2518051PFRNDFD). Swabs were also opened in the lab but not touched to any surfaces to function as negative controls. Samples and controls were processed inside a class II biosafety cabinet to avoid inadvertent cross contamination. The swabs were suspended in 15 ml of PBS (Phosphate Buffered Saline) and vortexed for 20 seconds to remove cells from the swab surface. The PBS was used to inoculate Petri dishes filled with TSA (Tryptic Soy Agar), Blood Agar, or Reasoners 2 agar to check for microbial growth. Each plate was inoculated with 0.1 ml of PBS. The TSA and blood agar plates were incubated at 35˚C and the Reasoners 2 agar plates were incubated at 25˚C for seven days. Petri dishes filled with Potato dextrose agar, Saboraud dextrose agar, or Saboraud dextrose agar with 0.1 mg/ml of chloramphenicol, an antibiotic, were used to check for fungal growth. These plates were inoculated with 0.3 ml of PBS and incubated at 30˚C. The remaining PBS was frozen at -80 ˚C for DNA sequencing. After incubation, isolates were counted and reisolated for identification. Isolates were identified using the VITEK23 system or by sequencing a portion of the 16S rRNA gene for bacteria or the ribosomal internal transcribed spacer (ITS) for fungi. Sequencing was performed with an ABI 3500 Sanger sequencer. Results: During our initial sampling, six of the seven sites sampled (86%) displayed bacterial or fungal growth. Samples collected from the staging areas and microtomy labs are not included in this calculation since those areas are maintained at a lower ISO 7 equivalent cleanliness standard. A month later, only three of the seven sites (43%) displayed bacterial growth. No fungal growth was detected in the second sampling. Since new equipment had been introduced to the Hayabusa2 lab since the first round of sampling, an additional three sampling sites were included in the second round of sampling. None of these sites displayed microbial growth. These sites will be included in all future sampling efforts. Bacterial isolates have been identified from the following genera at multiple time points: Micrococcus, Staphylococcus, and Bacillus. Isolates from the genera: Microbacterium, Nocardioides, Methylocystis, and Microvirga were identified in the initial sampling, but were not present at later time points. Identification of fungal isolates is in progress. Results are summarized in Table 1. Discussion: The recovery rate or percentage of positive samples4 was initially 86%, which is higher than the median recovery rate for comparable ISO 5 equivalent curation labs like Stardust (33%), Hayabusa (33%), and Cosmic Dust (50%). However, after a month of operation, the recovery rate for these same sites decreased to 43%, which is similar to what we observe in comparable curation cleanrooms with no microbial control requirements. Adding in the new sampling sites further decreases the recovery rate to 30%. With the reduction in recovery rate, we also observed a decrease in microbial diversity. At the first time point, we observed at least 10 different bacterial species and at least two different fungi. This is a higher diversity than the median values for comparable ISO 5 equivalent labs (2-4 isolates per sampling event). After the second sampling, we observed at least 4 bacterial species and no fungi, which is more consistent with comparable labs. We expect the recovery rate and diversity in both labs to continue to decrease as routine operation continues. We will use ultrapure hydrogen peroxide to disinfect equipment and work areas prior to opening any sample containers. Most of the bacterial and fungal isolates were detected on samples from the cleanroom floors. This is consistent with baseline results from other curation labs. Organisms from the genera Bacillus, Staphylococcus, and Micrococcus that were repeatedly detected are common in cleanrooms and on human skin5,6. These organisms are generally thought to be introduced when people enter the cleanroom. Microbacterium, Nocardioides, and Microvirga have also previously been identified in astromaterials cleanrooms, but not as frequently as Bacillus, Staphylococcus, and Micrococcus. Methylocystis is a novel genus in the astromaterials cleanrooms, but it was identified with low accuracy (93% match in the sequenced region of the 16S rRNA gene) and further work is needed to confirm this identification. Microbacterium is a diverse genus with isolates identified from terrestrial and aquatic sediments. Some species of Microbacterium are capable of degrading complex organic compounds found in crude oil. The presence of these bacteria in the OSIRIS REx and Hayabusa2 cleanrooms should be closely monitored. Methylocystis is a genus of methanotrophic bacteria capable of oxidizing methane. If this identification proves to be correct and it is detected again, it should be closely monitored as well. Under nominal operating conditions, samples should not ever encounter the cleanroom floor or other high traffic areas. If we observe an increase in the bioburden in sensitive work areas that appears to be influenced by organism transfer from high traffic areas like the floors, we can employ additional hydrogen peroxide treatments to disinfect high traffic areas. Routine microbial monitoring of these labs will ensure that NASA’s astromaterials collections remain pristine and useful for scientific study. Table 1. Sampling Locations and Colony Counts Bacterial CFUa Fungal CFU Bacterial CFU Fungal CFU Lab - Location 11/2/2021 11/2/2021 12/13/2021 12/13/2021 H2b-Floor 4 8 1 0 H2-staging pass through 3 0 0 0 H2-microtomy pass through TNTCc 0 0 0 H2 Microscope 1 NA NA 0 0 H2 Microscope 2 NA NA 0 0 H2-Table NA NA 0 0 OREXd- microtomy pass through 0 0 6 0 OREX – Anteroom pass through 0 0 0 0 OREX – Floor 1 2 0 0 OREX Witness Foil Table 3 0 1 0 Staging-Floor 16 0 15 0 Microtomy-Floor 3 0 2 0 a: CFU = Colony Forming Unit b: H2 = Hayabusa2 Lab c: TNTC = too numerous to count d: OREX = OSIRIS-REx Lab References: 1. ISO 14644-1:2015 - Cleanrooms and associated controlled environments -- Part 1: Classification of air cleanliness by particle concentration. 37 (2015). 2. McCubbin, F. M. et al. Space Sci Rev 215, (2019). 3. Pincus, D. H. Encyclopedia of Rapid Microbiological Methods (2005). 4. The United States Pharmacopeial Convention. USP General Chapter <1116> 17, 784–794 (2013). 5. Sheraba, N. S., Yassin, A. S. & Amin, M. BMC Research Notes 3, 278 (2010). 6. Utescher, C. L. de A., Franzolin, M. R., Trabulsi, L. R. & Gambale, V. Brazilian Journal of Microbiology 38, 710–716 (2007).

A B Regberg↗
NASA NTRS2013

Advanced Curation of Current and Future Extraterrestrial Samples

Curation of extraterrestrial samples is the critical interface between sample return missions and the international research community. Curation includes documentation, preservation, preparation, and distribution of samples. The current collections of extraterrestrial samples include: Lunar rocks / soils collected by the Apollo astronauts Meteorites, including samples of asteroids, the Moon, and Mars "Cosmic dust" (asteroid and comet particles) collected by high-altitude aircraft Solar wind atoms collected by the Genesis spacecraft Comet particles collected by the Stardust spacecraft Interstellar dust collected by the Stardust spacecraft Asteroid particles collected by the Hayabusa spacecraft These samples were formed in environments strikingly different from that on Earth. Terrestrial contamination can destroy much of the scientific significance of many extraterrestrial materials. In order to preserve the research value of these precious samples, contamination must be minimized, understood, and documented. In addition the samples must be preserved - as far as possible - from physical and chemical alteration. In 2011 NASA selected the OSIRIS-REx mission, designed to return samples from the primitive asteroid 1999 RQ36 (Bennu). JAXA will sample C-class asteroid 1999 JU3 with the Hayabusa-2 mission. ESA is considering the near-Earth asteroid sample return mission Marco Polo-R. The Decadal Survey listed the first lander in a Mars sample return campaign as its highest priority flagship-class mission, with sample return from the South Pole-Aitken basin and the surface of a comet among additional top priorities. The latest NASA budget proposal includes a mission to capture a 5-10 m asteroid and return it to the vicinity of the Moon as a target for future sampling. Samples, tools, containers, and contamination witness materials from any of these missions carry unique requirements for acquisition and curation. Some of these requirements represent significant advances over methods currently used. New analytical and screening techniques will increase the value of current sample collections. Improved web-based tools will make information on all samples more accessible to researchers and the public. Advanced curation of current and future extraterrestrial samples includes: Contamination Control - inorganic / organic Temperature of preservation - subfreezing / cryogenic Non-destructive preliminary examination - X-ray tomography / XRF mapping / Raman mapping Microscopic samples - handling / sectioning / transport Special samples - unopened lunar cores Informatics - online catalogs / community-based characterization.

Allen, Carlton C.↗
NASA NTRS2001

Mars Sample Handling Protocol Workshop Series: Workshop 2

Numerous NASA reports and studies have identified Planetary Protection (PP) as an important part of any Mars sample return mission. The mission architecture, hardware, on-board experiments, and related activities must be designed in ways that prevent both forward- and back-contamination and also ensure maximal return of scientific information. A key element of any PP effort for sample return missions is the development of guidelines for containment and analysis of returned sample(s). As part of that effort, NASA and the Space Studies Board (SSB) of the National Research Council (NRC) have each assembled experts from a wide range of scientific fields to identify and discuss issues pertinent to sample return. In 1997, the SSB released its report on recommendations for handling and testing of returned Mars samples. In particular, the NRC recommended that: a) samples returned from Mars by spacecraft should be contained and treated as potentially hazardous until proven otherwise, and b) rigorous physical, chemical, and biological analyses [should] confirm that there is no indication of the presence of any exogenous biological entity. Also in 1997, a Mars Sample Quarantine Protocol workshop was convened at NASA Ames Research Center to deal with three specific aspects of the initial handling of a returned Mars sample: 1) biocontainment, to prevent 'uncontrolled release' of sample material into the terrestrial environment; 2) life detection, to examine the sample for evidence of organisms; and 3) biohazard testing, to determine if the sample poses any threat to terrestrial life forms and the Earth's biosphere. In 1999, a study by NASA's Mars Sample Handling and Requirements Panel (MSHARP) addressed three other specific areas in anticipation of returning samples from Mars: 1) sample collection and transport back to Earth; 2) certification of the samples as non-hazardous; and 3) sample receiving, curation, and distribution. To further refine the requirements for sample hazard testing and the criteria for subsequent release of sample materials from quarantine, the NASA Planetary Protection Officer convened an additional series of workshops beginning in March 2000. The overall objective of these workshops was to develop comprehensive protocols to assess whether the returned materials contain any biological hazards, and to safeguard the purity of the samples from possible terrestrial contamination. This document is the report of the second Workshop in the Series. The information herein will ultimately be integrated into a final document reporting the proceedings of the entire Workshop Series along with additional information and recommendations.

Rummel, John D.↗
NASA NTRS2016

Preserving, Enhancing, and Continuing the Scientific Legacy of the Apollo Sample Suite

From 1969 to 1972, Apollo astronauts collected 382 kg of rocks, soils, and core samples from six geologically diverse locations on the Moon. In the nearly 50 years since the samples were collected, over 3000 different studies have been conducted using the nearly 2200 different Apollo samples. Despite the maturity of the sample collection, many new studies of lunar samples are undertaken each year, with an average of more than 55 requests and more than 600 distinct subsamples allocated annually over the past five years. The Apollo samples are a finite resource, however. Although new studies are encouraged, it is important that new studies do not duplicate previous studies, and where possible, leverage previous results to inform and enhance the current studies. This helps to preserve the samples and scientific funding, both of which are precious resources. We have initiated several new efforts to rescue some of the early analyses from these samples, including unpublished analytical data. We are actively scanning NASA documentation in paper form that is related to the Apollo missions and sample processing, and we are collaborating with IEDA to establish a geochemical data base called MoonDB. To populate this database, we are actively working with about a dozen prominent lunar PIs to organize and transcribe years of both published and unpublished data, making it available to all researchers. This effort will also take advantage of new online analytical tools like PetDB. There have already been tangible results from the MoonDB data rescue effort. A pilot project involving the rescue of geochemical data of John Delano on Apollo pyroclastic glasses has already been referenced in multiple Apollo sample requests, and in fact, the compiled data was used as part of one of the new studies. Similarly, scanned sample handling reports have been utilized to find previously analyzed samples that were appropriate to fulfill new sample requests. We have also begun to image the Apollo samples using (1) micro‐CT scanning to document the interior structure of samples, and (2) comprehensive high resolution photography of samples, enabling high resolution 3D reconstructions of the samples. Both efforts will provide comprehensive access to these samples and allow for more targeted requests, and thus better curation of the samples for decades to come.

Zeigler, Ryan↗
NASA NTRS2016

Spatial Sampling of Weather Data for Regional Crop Yield Simulations

Field-scale crop models are increasingly applied at spatio-temporal scales that range from regions to the globe and from decades up to 100 years. Sufficiently detailed data to capture the prevailing spatio-temporal heterogeneity in weather, soil, and management conditions as needed by crop models are rarely available. Effective sampling may overcome the problem of missing data but has rarely been investigated. In this study the effect of sampling weather data has been evaluated for simulating yields of winter wheat in a region in Germany over a 30-year period (1982-2011) using 12 process-based crop models. A stratified sampling was applied to compare the effect of different sizes of spatially sampled weather data (10, 30, 50, 100, 500, 1000 and full coverage of 34,078 sampling points) on simulated wheat yields. Stratified sampling was further compared with random sampling. Possible interactions between sample size and crop model were evaluated. The results showed differences in simulated yields among crop models but all models reproduced well the pattern of the stratification. Importantly, the regional mean of simulated yields based on full coverage could already be reproduced by a small sample of 10 points. This was also true for reproducing the temporal variability in simulated yields but more sampling points (about 100) were required to accurately reproduce spatial yield variability. The number of sampling points can be smaller when a stratified sampling is applied as compared to a random sampling. However, differences between crop models were observed including some interaction between the effect of sampling on simulated yields and the model used. We concluded that stratified sampling can considerably reduce the number of required simulations. But, differences between crop models must be considered as the choice for a specific model can have larger effects on simulated yields than the sampling strategy. Assessing the impact of sampling soil and crop management data for regional simulations of crop yields is still needed.

upscaling↗
NASA NTRS2022

Phase II and III Solid Waste Management Unit Assessment and Confirmatory Sampling Report for Per- and Polyfluorinated Alkyl Substances, Potential Release Location 237, Kennedy Space Center, Florida

Per- and polyfluoroalkyl substances (PFAS) Phase II and III Solid Waste Management Unit (SWMU) Assessment (SA) and Confirmatory Sampling (CS) were conducted at the John F. Kennedy Space Center (KSC), Potential Release Location (PRL) 237, located in Merritt Island, Brevard County, Florida for the National Aeronautics and Space Administration (NASA) under Contract 80KSC019D0010/80KSC019F0289. A Phase I SA and CS Report was previously submitted that identified 33 locations of concern (LOCs) at KSC associated with the storage, use, or release of materials containing PFAS. The objectives of the Phase II and III SA and CS were to: - Complete site and source characterization of identified LOCs; - Identify areas of potential concern (AOPCs) where past or present activities may have resulted in a release of PFAS to the environment; - Qualitatively characterize migration potential of released PFAS to environmental media (primarily soil, groundwater, surface water, and sediment); and - Identify potential receptors for exposure to PFAS-impacted environmental media. Surface water samples were collected from LOCs/AOPCs and locations on the KSC periphery during PFAS CS Phases I, II and III. Including duplicate samples, 143 environmental samples were analyzed for 14 PFAS compounds and 104 environmental samples were analyzed for 18 PFAS compounds. Ten of the analytes on the 14 compound list were detected at least once in surface water at KSC, with a wide range in magnitude of detections. The State of Florida has surface water screening levels (SWSLs) for perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) in surface water, both for human and ecological receptors. The human health screening levels are lower than the ecological ones and are used in this report. PFOA was not detected in surface water at concentrations exceeding the SWSL at any LOC or Area of Potential Concern (AOPC). PFOS was detected in surface water at concentrations exceeding the SWSL at 28 LOCs/AOPCs, plus 42 locations on the KSC periphery. The data suggest a wide distribution of PFAS, particularly PFOS, in surface water but the sources of surface water contamination are not yet adequately understood. Sediment samples were collected during Phases II and III but not Phase I. Including duplicate samples, 104 environmental samples were analyzed for 14 PFAS compounds and 10 environmental samples were analyzed for 18 PFAS compounds. Twelve compounds were detected in sediment at KSC, with a wide range in magnitude of detections. There are currently no screening levels or cleanup target levels (CTLs) for sediment. The need to delineate and further characterize PFAS in sediment will be evaluated for those sites that move forward into Site Assessment. Groundwater samples were collected via DPT from LOCs/AOPCs and locations on the KSC periphery during PFAS CS Phases I, II and III. Including duplicate samples, 812 environmental DPT groundwater samples were analyzed for 14 PFAS compounds and 247 environmental samples were analyzed for 18 PFAS compounds. Twelve of the 14 compounds were detected in groundwater at KSC, with a wide range in magnitude of detections. The State of Florida has provisional groundwater cleanup target levels (GCTLs) for PFOA, PFOS and the additive concentration of PFOA+PFOS. PFOA was detected in groundwater at concentrations exceeding its provisional GCTL at 16 LOCs/AOPCs, with no exceedances on the KSC periphery. PFOS was detected in groundwater at concentrations exceeding its provisional GCTL at 30 LOCs/AOPCs, plus two locations on the KSC periphery. PFOA+PFOS was detected in groundwater at concentrations exceeding the provisional GCTL at 31 LOCs/AOPCs, plus two locations on the KSC periphery. Soil samples were collected during Phases II and III but not Phase I. Including duplicate samples, 292 environmental samples were analyzed for 14 PFAS compounds and 80 environmental samples were analyzed for 18 PFAS compounds. Fourteen compounds were detected in soil at KSC, with a wide range in magnitude of detections. The State of Florida has provisional soil cleanup target levels (SCTLs) for PFOA and PFOS. There were no PFOA results above its provisional SCTL. There was one LOC with a result above the provisional SCTL for residential settings. Recommendations are presented for each LOC and AOPC.

PFAS↗
NASA NTRSDate not supplied

ISS External Microorganisms: Collecting Planetary Protection Samples During Extravehicular Activity

We have developed, tested, and flown a caddy capable of collecting aseptic samples from external surfaces of the ISS (International Space Station). The sampling caddy is certified for use during US EVA (extra vehicular activity) and was launched to ISS in the summer of 2023. We are scheduled to collect samples from 6 locations outside ISS during an EVA in May of 2024. We will freeze these samples at -80°C on orbit and return them to Earth. We will then extract and sequence any DNA collected during the EVA using next generation sequencing technologies to characterize the community composition and function of each sample. Measuring the type and quantity of microbes present on the exterior of ISS will allow us to address knowledge gap 2B, “Acceptable levels of microbial/organic releases from humans and support systems” described in a 2019 COSPAR report. Collecting data about microbial release from current crewed vehicles will inform requirements for acceptable leak rates for future crewed missions to Mars. The sampling kit consists of eight commercially available, sterile, DNA free, macrofoam swabs ( 23 mm. diameter ) installed in custom aluminum end effectors. Each end effector is housed in and individual aluminum canister. Each canister contains a 0.2 μm Teflon filter to allow the interior volume to accommodate pressure changes without permitting microbial contaminants to enter the sterile interior volume. A handle repurposed from the space shuttle tile repair kit is used to remove the end effector from the sample canister, collect a sample by swabbing a surface and then replace the end effector in its canister. The canisters and end effectors were cleaned and assembled on Earth. Prior to installing the sterile swab the canisters and end effectors were sterilized in an autoclave at 134°C, 215 kPA, for 7 min.. The final assembly occurred in a sterilized class II biosafety cabinet. We will collect six samples from the 1) airlock vestibule, 2) airlock thermal cover, 3) a gap in the micrometeorite shielding near the airlock, 4) a handrail near the airlock, 5) the CDRA (Carbon Dioxide Removal Assembly) vent, and 6) the VES (Vacuum Exhaust System) vent. The remaining two swabs will be reserved as controls. One swab will be exposed during the EVA without touching any surfaces to act as a blank. The final swab will remain sealed until the entire sampling kit is returned to earth. Based on previously published results from the Russian segment, we hypothesize that there will be detectable microbes at some or all of these locations. Ground-based testing of this sampling caddy confirms that the swabs remain sterile as the canisters transition in and out of vacuum. We were able to retrieve, viable bacterial and fungal cells as well as DNA from samples collected from US space suits during vacuum chamber tests lasting as long as seven hours. Based on these results and feedback from the test subjects the sampling caddy was modified to improve ergonomics and meet US EVA safety requirements. Bayonet probes were added to the sides of the sample kit as alternate mounting points. Additional locking features were added to the end effector and the filter stack to prevent inadvertent release during use. The opening mechanism was changed from one where the end effector was rocked laterally to defeat a ball detent to a twist-to-open threaded closure for similar reasons. Demonstrating, this sampling caddy’s effectiveness during a US EVA will allow us to address knowledge gaps identified in COSPAR reports and begin to define planetary protection requirements for life support systems on crewed missions to mars. This kit could also be used to collect contamination control samples during Artemis missions to verify requirements and could be easily modified for robotic sample collection.

Planetary Protection↗
NASA NTRS1988

Sampling Mars: Analytical requirements and work to do in advance

Sending a mission to Mars to collect samples and return them to the Earth for analysis is without doubt one of the most exciting and important tasks for planetary science in the near future. Many scientifically important questions are associated with the knowledge of the composition and structure of Martian samples. Amongst the most exciting questions is the clarification of the SNC problem- to prove or disprove a possible Martian origin of these meteorites. Since SNC meteorites have been used to infer the chemistry of the planet Mars, and its evolution (including the accretion history), it would be important to know if the whole story is true. But before addressing possible scientific results, we have to deal with the analytical requirements, and with possible pre-return work. It is unlikely to expect that a possible Mars sample return mission will bring back anything close to the amount returned by the Apollo missions. It will be more like the amount returned by the Luna missions, or at least in that order of magnitude. This requires very careful sample selection, and very precise analytical techniques. These techniques should be able to use minimal sample sizes and on the other hand optimize the scientific output. The possibility to work with extremely small samples should not obstruct another problem: possible sampling errors. As we know from terrestrial geochemical studies, sampling procedures are quite complicated and elaborate to ensure avoiding sampling errors. The significance of analyzing a milligram or submilligram sized sample and putting that in relationship with the genesis of whole planetary crusts has to be viewed with care. This leaves a dilemma on one hand, to minimize the sample size as far as possible in order to have the possibility of returning as many different samples as possible, and on the other hand to take a sample large enough to be representative. Whole rock samples are very useful, but should not exceed the 20 to 50 g range, except in cases of extreme inhomogeneity, because for larger samples the information tends to become redundant. Soil samples should be in the 2 to 10 g range, permitting the splitting of the returned samples for studies in different laboratories with variety of techniques.

Koeberl, Christian↗