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At least 199 records · Page 11

IMPACT-ing Exploration Spaceflight Risk Prediction and Medical System Design

Human exploration spaceflight missions to the Moon and Mars present unprecedented challenges for in-mission medical care. Compared with the ISS, the greater distance from Earth will mean increased mission durations, communication delays, limited to no resupply opportunities, and significant limitations on the evacuation of ill or injured crew. Spacecraft mass, volume, and power will be curtailed while higher demands will be placed on the crew’s knowledge, skills, and abilities. In this higher risk environment, it is important to: a) quantitatively estimate human system risk attributable to medical conditions, a process known as Probabilistic Risk Analysis, and b) use these estimates to inform medical system design. IMPACT (Informing Mission Planning via Analysis of Complex Tradespaces) is a PRA and medical trade space analysis tool developed by NASA to advance exploration mission medical system design. IMPACT improves upon and will soon replace NASA’s existing tool, the Integrated Medical Model, with: a novel evidence base baselined to exploration environments; an expanded list of 120 medical conditions; a significant increase in the number of medical resources that can be utilized and in the flexibility of their use; and the modelling of time lost performing mission-specific tasks due to medical conditions. IMPACT provides evidence-based, mission-specific PRA estimates of in-flight medical risk and an initial list of clinical capabilities and medical resources/hardware to be considered. In addition, sophisticated trade space capabilities estimate how human system risk varies with changes to the mission architecture or medical capability set (e.g., if the system mass constraint decreases by 10% or ultrasound is removed). This panel will provide an overview of IMPACT, its intended use cases, and future development plans. The panel will be the first public presentation of IMPACT results, including medical risk estimates for extended duration Artemis missions, the medical conditions most influencing medical risk metrics, and the clinical capabilities that have the largest effect on medical risk.

Ben Easter↗

Preliminary Medical Risk Estimates and Clinical Capability Needs for Artemis IV

BACKGROUND Human exploration spaceflight missions to the Moon and Mars present unprecedented challenges for in- mission medical care. Compared with the ISS, the greater distance from Earth will mean increased mission durations, communication delays, limited to no resupply opportunities, and significant limitations on the evacuation of ill or injured crew. Spacecraft mass, volume, and power will be curtailed while higher demands will be placed on the crew’s knowledge, skills, and abilities. In this higher risk environment, it is important to: a) quantitatively estimate human system risk attributable to medical conditions, a process known as Probabilistic Risk Analysis, and b) use these estimates to inform medical system design. IMPACT (Informing Mission Planning via Analysis of Complex Tradespaces) is a PRA and medical trade space analysis tool developed by NASA to advance exploration mission medical system design. IMPACT v1.0 improves upon and will soon replace NASA’s existing tool, the Integrated Medical Model, with: a novel evidence base baselined to exploration environments; an expanded list of 119 medical conditions; a significant increase in the number of medical resources that can be utilized and in the flexibility of their use; and the modelling of time lost performing mission-specific tasks due to medical conditions. METHODOLOGY: This abstract will present IMPACT estimates of medical system risk and clinical capability needs for the Artemis IV mission. Artemis IV is currently scheduled for 2026 and will visit the Gateway space station in lunar orbit prior to the second lunar landing of the Artemis program. The baseline Artemis IV mission that was modeled was 28 days in duration with phases including Orion outbound, 4 days on the Gateway space station in lunar orbit, 2 crew on the surface of the Moon for approximately one week, an additional 5 days on Gateway, and then return to Earth. This baseline was compared to two alternative 34-day design reference missions (DRMs) that shifted the lunar sortie earlier or later in the mission profile. Assumptions included 2 female and 2 male crew and a notional medical system mass of 25 kg. Medical system risk estimates include loss of crew life (LOCL), consideration of medical evacuation (known as return to definitive care – RTDC), and an estimate of crew time lost due to medical conditions (Task Time Lost – TTL). The presentation will also describe the medical conditions that are the greatest drivers of risk as well as the clinical capabilities and resources that have the largest effect on risk. RESULTS: All three DRMs had very low probability of LOCL from medical conditions, primarily due to short duration. RTDC was also similar across the DRMs. In contrast, TTL was higher in the early lunar sortie DRM due to earlier occurrence of EVA-related medical conditions. Taken as a whole, there was no clinically significant difference in medical risk across the three missions. Results for clinical capabilities and an example medical equipment list will be discussed but were similar across DRMs.

D Hilmers↗

Medical Risk Estimates and Clinical Capability Needs for A Long Duration Artemis Mission

BACKGROUND Human exploration spaceflight missions to the Moon and Mars present unprecedented challenges for in-mission medical care. Compared with the ISS, the greater distance from Earth will mean increased mission durations, communication delays, limited to no resupply opportunities, and significant limitations on the evacuation of ill or injured crew. Spacecraft mass, volume, and power will be curtailed while higher demands will be placed on the crew’s knowledge, skills, and abilities. In this higher risk environment, it is important to: a) quantitatively estimate human system risk attributable to medical conditions, a process known as Probabilistic Risk Analysis, and b) use these estimates to inform medical system design. IMPACT (Informing Mission Planning via Analysis of Complex Tradespaces) is a PRA and medical trade space analysis tool developed by NASA to advance exploration mission medical system design. IMPACT v1.0 improves upon and will soon replace NASA’s existing tool, the Integrated Medical Model, with: a novel evidence base baselined to exploration environments; an expanded list of 119 medical conditions; a significant increase in the number of medical resources that can be utilized and in the flexibility of their use; and the modelling of time lost performing mission-specific tasks due to medical conditions. DISCUSSION: This abstract will present IMPACT estimates of medical system risk and clinical capability needs for an extended duration Artemis mission of approximately 9 months, with phases including outbound transit, 3 months on the Lunar Gateway space station, 3 months on the Lunar surface with EVAs, 3 months on Gateway (simulating the return phase of a Mars mission), and transit back to Earth. Medical system risk estimates include loss of crew life (LOCL), consideration of medical evacuation (known as return to definitive care – RTDC), and an estimate of crew time lost due to medical conditions (Task Time Lost – TTL). The presentation will also describe the medical conditions that are the greatest drivers of risk as well as the clinical capabilities and resources that have the largest effect on risk.

W Thompson↗

Medical Risk Estimates and Clinical Capability Needs for a Late Artemis Mission

BACKGROUND Human exploration spaceflight missions to the Moon and Mars present unprecedented challenges for in-mission medical care. Compared with the ISS, the greater distance from Earth will mean increased mission durations, communication delays, limited to no resupply opportunities, and significant limitations on the evacuation of ill or injured crew. Spacecraft mass, volume, and power will be curtailed while higher demands will be placed on the crew’s knowledge, skills, and abilities. In this higher risk environment, it is important to: a) quantitatively estimate human system risk attributable to medical conditions, a process known as Probabilistic Risk Analysis, and b) use these estimates to inform medical system design. IMPACT (Informing Mission Planning via Analysis of Complex Tradespaces) is a PRA and medical trade space analysis tool developed by NASA to advance exploration mission medical system design. IMPACT v1.0 improves upon and will soon replace NASA’s existing tool, the Integrated Medical Model, with: a novel evidence base baselined to exploration environments; an expanded list of 119 medical conditions; a significant increase in the number of medical resources that can be utilized and in the flexibility of their use; and the modelling of time lost performing mission-specific tasks due to medical conditions. DISCUSSION: This abstract will present IMPACT estimates of medical system risk and clinical capability needs for an extended duration Artemis mission of approximately 9 months, with phases including outbound transit, 3 months on the Lunar Gateway space station, 3 months on the Lunar surface with EVAs, 3 months on Gateway (simulating the return phase of a Mars mission), and transit back to Earth. Medical system risk estimates include loss of crew life (LOCL), consideration of medical evacuation (known as return to definitive care –RTDC), and an estimate of crew time list due to medical conditions (Task Time Lost –TTL). The presentation will also describe the medical conditions that are the greatest drivers of risk as well as the clinical capabilities and resources that have the largest effect on risk.

A Anderson↗

Preliminary Medical Risk Estimates and Clinical Capability Needs for Late Artemis Missions

BACKGROUND Human exploration spaceflight missions to the Moon and Mars present unprecedented challenges for in- mission medical care. Compared with the ISS, the greater distance from Earth will mean increased mission durations, communication delays, limited to no resupply opportunities, and significant limitations on the evacuation of ill or injured crew. Spacecraft mass, volume, and power will be curtailed while higher demands will be placed on the crew’s knowledge, skills, and abilities. In this higher risk environment, it is important to: a) quantitatively estimate human system risk attributable to medical conditions, a process known as Probabilistic Risk Analysis, and b) use these estimates to inform medical system design. IMPACT (Informing Mission Planning via Analysis of Complex Tradespaces) is a PRA and medical trade space analysis tool developed by NASA to advance exploration mission medical system design. IMPACT v1.0 improves upon and will soon replace NASA’s existing tool, the Integrated Medical Model, with: a novel evidence base baselined to exploration environments; an expanded list of 119 medical conditions; a significant increase in the number of medical resources that can be utilized and in the flexibility of their use; and the modelling of time lost performing mission-specific tasks due to medical conditions. METHODOLOGY: This abstract will present IMPACT estimates of medical system risk and clinical capability needs for the Artemis IV mission. Artemis IV is currently scheduled for 2026 and will visit the Gateway space station in lunar orbit prior to the second lunar landing of the Artemis program. The baseline Artemis IV mission that was modeled was 28 days in duration with phases including Orion outbound, 4 days on the Gateway space station in lunar orbit, 2 crew on the surface of the Moon for approximately one week, an additional 5 days on Gateway, and then return to Earth. This baseline was compared to two alternative 34-day design reference missions (DRMs) that shifted the lunar sortie earlier or later in the mission profile. Assumptions included 2 female and 2 male crew and a notional medical system mass of 25 kg. Medical system risk estimates include loss of crew life (LOCL), consideration of medical evacuation (known as return to definitive care – RTDC), and an estimate of crew time lost due to medical conditions (Task Time Lost – TTL). The presentation will also describe the medical conditions that are the greatest drivers of risk as well as the clinical capabilities and resources that have the largest effect on risk. RESULTS: All three DRMs had very low probability of LOCL from medical conditions, primarily due to short duration. RTDC was also similar across the DRMs. In contrast, TTL was higher in the early lunar sortie DRM due to earlier occurrence of EVA-related medical conditions. Taken as a whole, there was no clinically significant difference in medical risk across the three missions. Results for clinical capabilities and an example medical equipment list will be discussed but were similar across DRMs.

D. Hilmers↗

Agent Based Software for the Autonomous Control of Formation Flying Spacecraft

Distributed satellite systems is an enabling technology for many future NASA/DoD earth and space science missions, such as MMS, MAXIM, Leonardo, and LISA [1, 2, 3]. While formation flying offers significant science benefits, to reduce the operating costs for these missions it will be essential that these multiple vehicles effectively act as a single spacecraft by performing coordinated observations. Autonomous guidance, navigation, and control as part of a coordinated fleet-autonomy is a key technology that will help accomplish this complex goal. This is no small task, as most current space missions require significant input from the ground for even relatively simple decisions such as thruster burns. Work for the NMP DS1 mission focused on the development of the New Millennium Remote Agent (NMRA) architecture for autonomous spacecraft control systems. NMRA integrates traditional real-time monitoring and control with components for constraint-based planning, robust multi-threaded execution, and model-based diagnosis and reconfiguration. The complexity of using an autonomous approach for space flight software was evident when most of its capabilities were stripped off prior to launch (although more capability was uplinked subsequently, and the resulting demonstration was very successful).

How, Jonathan P.↗

Advanced Integration Matrix Education Outreach

The Advanced Integration Matrix (AIM) will design a ground-based test facility for developing revolutionary integrated systems for joint human-robotic missions in order to study and solve systems-level integration issues for exploration missions beyond Low Earth Orbit (LEO). This paper describes development plans for educational outreach activities related to technological and operational integration scenarios similar to the challenges that will be encountered through this project. The education outreach activities will provide hands-on, interactive exercises to allow students of all levels to experience design and operational challenges similar to what NASA deals with everyday in performing the integration of complex missions. These experiences will relate to and impact students everyday lives by demonstrating how their interests in science and engineering can develop into future careers, and reinforcing the concepts of teamwork and conflict resolution. Allowing students to experience and contribute to real-world development, research, and scientific studies of ground-based simulations for complex exploration missions will stimulate interest in the space program, and bring NASA's challenges to the student level. By enhancing existing educational programs and developing innovative activities and presentations, AIM will support NASA s endeavor to "inspire the next generation of explorers.. .as only NASA can."

Paul Heather L.↗

Tool Use Within NASA Software Quality Assurance

As space mission software systems become larger and more complex, it is increasingly important for the software assurance effort to have the ability to effectively assess both the artifacts produced during software system development and the development process itself. Conceptually, assurance is a straightforward idea - it is the result of activities carried out by an organization independent of the software developers to better inform project management of potential technical and programmatic risks, and thus increase management's confidence in the decisions they ultimately make. In practice, effective assurance for large, complex systems often entails assessing large, complex software artifacts (e.g., requirements specifications, architectural descriptions) as well as substantial amounts of unstructured information (e.g., anomaly reports resulting from testing activities during development). In such an environment, assurance engineers can benefit greatly from appropriate tool support. In order to do so, an assurance organization will need accurate and timely information on the tool support available for various types of assurance activities. In this paper, we investigate the current use of tool support for assurance organizations within NASA, and describe on-going work at JPL for providing assurance organizations with the information about tools they need to use them effectively.

software assurance↗

Mind the Gap: Addressing Data Gaps and Assessing Noise Mismodeling in LISA

Due to the sheer complexity of the Laser Interferometer Space Antenna (LISA) space mission, data gaps arising from instrumental irregularities and/or scheduled maintenance are unavoidable. Focusing on merger-dominated massive black hole binary signals, we test the appropriateness of the Whittle-likelihood on gapped data in a variety of cases. From first principles, we derive the likelihood valid for gapped data in both the time and frequency domains. Cheap-to-evaluate proxies to p-p plots are derived based on a Fisher-based formalism, and verified through Bayesian techniques. Our tools allow to predict the altered variance in the parameter estimates that arises from noise mismodeling, as well as the information loss represented by the broadening of the posteriors. The result of noise mismodeling with gaps is sensitive to the characteristics of the noise model, with strong low-frequency (red) noise and strong high-frequency (blue) noise giving statistically significant fluctuations in recovered parameters. We demonstrate that the introduction of a tapering window reduces statistical inconsistency errors, at the cost of less precise parameter estimates. We also show that the assumption of independence between inter-gap segments appears to be a fair approximation even if the data set is inherently coherent. However, if one instead assumes fictitious correlations in the data stream, when the data segments are actually independent, then the resultant parameter recoveries could be inconsistent with the true parameters. The theoretical and numerical practices that are presented in this work could readily be incorporated into global-fit pipelines operating on gapped data.

LISA↗

Technical Reference Suite Addressing Challenges of Providing Assurance for Fault Management Architectural Design

Research into complexities of software systems Fault Management (FM) and how architectural design decisions affect safety, preservation of assets, and maintenance of desired system functionality has coalesced into a technical reference (TR) suite that advances the provision of safety and mission assurance. The NASA Independent Verification and Validation (IVV) Program, with Software Assurance Research Program support, extracted FM architectures across the IVV portfolio to evaluate robustness, assess visibility for validation and test, and define software assurance methods applied to the architectures and designs. This investigation spanned IVV projects with seven different primary developers, a wide range of sizes and complexities, and encompassed Deep Space Robotic, Human Spaceflight, and Earth Orbiter mission FM architectures. The initiative continues with an expansion of the TR suite to include Launch Vehicles, adding the benefit of investigating differences intrinsic to model-based FM architectures and insight into complexities of FM within an Agile software development environment, in order to improve awareness of how nontraditional processes affect FM architectural design and system health management.

Fitz, Rhonda↗

Autonomy and the human element in space

NASA is contemplating the next logical step in the U.S. space program - the permanent presence of humans in space. As currently envisioned, the initial system, planned for the early 1990's, will consist of manned and unmanned platforms situated primarily in low Earth orbit. The manned component will most likely be inhabited by 6-8 crew members performing a variety of tasks such as materials processing, satellite servicing, and life science experiments. The station thus has utility in scientific and commercial enterprises, in national security, and in the development of advanced space technology. The technical foundations for this next step have been firmly established as a result of unmanned spacecraft missions to other planets, the Apollo program, and Skylab. With the shuttle, NASA inaugurates a new era of frequent flights and more routine space operations supporting a larger variety of missions. A permanently manned space system will enable NASA to expand the scope of its activities still further. Since NASA' s inception there has been an intense debate over the relative merits of manned and unmanned space systems. Despite the generally higher costs associated with manned components, astronauts have accomplished numerous essential, complex tasks in space. The unique human talent to evaluate and respond inventively to unanticipated events has been crucial in many missions, and the presence of crews has helped arouse and sustain public interest in the space program. On the other hand, the hostile orbital environment affects astronaut physiology and productivity, is dangerous, and mandates extensive support systems. Safety and cost factors require the entire station complex, both space and ground components, to be highly automated to free people from mundane operational chores. Recent advances in computer technology, artificial intelligence (AI), and robotics have the potential to greatly extend space station operations, offering lower costs and superior productivity. Extended operations can in turn enhance critical technologies and contribute to the competitive economic abilities of the United States. A high degree of automation and autonomy may be required to reduce dependence on ground systems, reduce mission costs, diminish complexity as perceived by the crew, increase mission lifetime and expand mission versatility. However, technologies dealing with heavily automated, long duration habitable spacecraft have not yet been thoroughly investigated by NASA. A highly automated station must amalgamate the diverse capabilities of people, machines, and computers to yield an efficient system which capitalizes on unique human characteristics. The station also must have an initial design which allows evolution to a larger and more sophisticated space presence. In the early years it is likely that AI-based subsystems will be used primarily in an advisory or planning capacity. As human confidence in automated systems grows and as technology advances, machines will take on more critical and interdependent roles. The question is whether, and how much, system autonomy will lead to improved station effectiveness.

Source record↗

Multimission Software Reuse in an Environment of Large Paradigm Shifts

The ground data systems provided for NASA space mission support are discussed. As space missions expand, the ground systems requirements become more complex. Current ground data systems provide for telemetry, command, and uplink and downlink processing capabilities. The new millennium project (NMP) technology testbed for 21st century NASA missions is discussed. The program demonstrates spacecraft and ground system technologies. The paradigm shift from detailed ground sequencing to a goal oriented planning approach is considered. The work carried out to meet this paradigm for the Deep Space-1 (DS-1) mission is outlined.

Wilson, Robert K.↗

Technical Reference Suite Addressing Challenges of Providing Assurance for Fault Management Architectural Design

Research into complexities of software systems Fault Management (FM) and how architectural design decisions affect safety, preservation of assets, and maintenance of desired system functionality has coalesced into a technical reference (TR) suite that advances the provision of safety and mission assurance. The NASA Independent Verification and Validation (IV&V) Program, with Software Assurance Research Program support, extracted FM architectures across the IV&V portfolio to evaluate robustness, assess visibility for validation and test, and define software assurance methods applied to the architectures and designs. This investigation spanned IV&V projects with seven different primary developers, a wide range of sizes and complexities, and encompassed Deep Space Robotic, Human Spaceflight, and Earth Orbiter mission FM architectures. The initiative continues with an expansion of the TR suite to include Launch Vehicles, adding the benefit of investigating differences intrinsic to model-based FM architectures and insight into complexities of FM within an Agile software development environment, in order to improve awareness of how nontraditional processes affect FM architectural design and system health management. The identification of particular FM architectures, visibility, and associated IV&V techniques provides a TR suite that enables greater assurance that critical software systems will adequately protect against faults and respond to adverse conditions. Additionally, the role FM has with regard to strengthened security requirements, with potential to advance overall asset protection of flight software systems, is being addressed with the development of an adverse conditions database encompassing flight software vulnerabilities. Capitalizing on the established framework, this TR suite provides assurance capability for a variety of FM architectures and varied development approaches. Research results are being disseminated across NASA, other agencies, and the software community. This paper discusses the findings and TR suite informing the FM domain in best practices for FM architectural design, visibility observations, and methods employed for IV&V and mission assurance.

Fitz, Rhonda↗

The Importance of Technology Readiness in NASA Earth Venture Missions

The first set of Venture-class investigations share the characteristic that the technology should be mature and all investigations must use mature technology that has been modeled or demonstrated in a relevant environment (Technology Readiness Level (TRL) >5). Technology Readiness Levels are a systematic metric/measurement system that supports assessments of the maturity of a particular technology and the consistent comparison of maturity between different types of technology. The TRL is used in NASA technology planning. A major step in the level of fidelity of the technology demonstration follows the completion of TRL 5. At TRL 6, a system or subsystem model or prototype must be demonstrated in a relevant environment (ground or space) representative model or prototype system or system, which would go well beyond ad hoc, "patch-cord," or discrete component level breadboarding. These TRL levels are chosen as target objectives for the Program. The challenge for offerors is that they must identify key aspects (uncertainty, multi subsystem complexity, etc) of the TRL estimate that should be properly explained in a submitted proposal. Risk minimization is a key component of the Earth Venture missions. Experiences of prior airborne missions will be shared. The discussion will address aspects of uncertainty and issues surrounding three areas of airborne earth science missions: (1) Aircraft or proposed flight platform -- Expressing the capability of the aircraft in terms of the supporting mission requirements. These issues include airplane performance characteristics (duration, range, altitude, among others) and multiship complexities. (2) Instruments -- Establishing that the instruments have been demonstrated in a relevant environment. Instruments with heritage in prior space missions meet this requirement, as do instruments tested on the ground. Evidence that the instruments have demonstrated the ability to collect data as advertised will be described. The complexity of the integration of multiple subsystems will also be addressed. Issues associated with tailoring the instrument to meet the specific Venture mission objectives must be thoroughly explained and justified. (3) Aircraft/Instrument Integration -- Explicitly defining what development may be required to harden the instrument and integrate the instrument. The challenges associated with this key aspect of major airborne earth science investigations will be presented.

Wells, James E.↗

Architecture Modeling and Performance Characterization of Space Communications and Navigation (SCaN) Network Using MACHETE

As future space exploration missions will involve larger number of spacecraft and more complex systems, theoretical analysis alone may have limitations on characterizing system performance and interactions among the systems. Simulation tools can be useful for system performance characterization through detailed modeling and simulation of the systems and its environment...This paper reports the simulation of the Orion (Crew Exploration Vehicle) to the International Space Station (ISS) mission where Orion is launched by Ares into orbit on a 14-day mission to rendezvous with the ISS. Communications services for the mission are provided by the Space Communication and Navigation (SCaN) network infrastructure which includes the NASA Space Network (SN), Ground Network (GN) and NASA Integrated Services Network (NISN). The objectives of the simulation are to determine whether SCaN can meet the communications needs of the mission, to demonstrate the benefit of using QoS prioritization, and to evaluate network-key parameters of interest such as delay and throughout.

space based networking↗

Toward intelligent robot systems in aerospace

The incorporation of progressively more autonomous capabilities in spacecraft has been made possible by advancements in electronics technologies for sensors, communication, and computing equipment; as a result, space missions have been able to cope with ever-increasing complexity and data throughputs, as demonstrated by the six-order-of-magnitude increase in planetary mission data rates. In order to continue this pace of development into the Space Station era, NASA has accelerated its R&D in automation and robotics, with emphasis on autonomous, knowledge-based and expert system-employing technologies and AI.

Heer, Ewald↗

Shuttle/Centaur Upper Stage Capability

A joint project to design, develop, procure, and produce Centaur upper stages for use with the Space Shuttle is discussed. A common Centaur G stage 6 meters (19.5 feet) in length is being jointly developed. A longer version designated Centaur G Prime is being developed by NASA to accomplish the Galileo and International Solar-Polar Mission flights in 1986. The Centaur G and G Prime will have the capability to place, respectively, approximately 4540 kilograms (10,000 pounds) and 5910 kilograms (13,000 pounds) into geosynchronous orbit from a standard Shuttle parking orbit of 278 kilometers (150 nautical miles) and Shuttle performance (lift) capability 29,500 kilograms (65,000 pounds). The advent of high energy upper stage capability in 1986 will permit space users and spacecraft developers to utilize spacecraft growth, stage combination concepts with storage modules, teleoperator systems, and other mission peculiar devices to satisfy complex mission demands. These capabilities should greatly enhance the usefulness of the space environment and stimulate mission planners toward conception of innovative means to meet ever increasing mission requirements.

Clark, H. J.↗

Benchmarking Computational Tools for Calling SNPs and Indels in Complex Microbial Populations

The NASA BioNutrients missions seek to understand the suitability of microorganisms for bioproduction during space flight. One topic of interest is the stability of microbial genomes during long-term ambient storage and subsequent rehydration and growth. To address these questions, samples from 8 species were flown to ISS for 5 years of desiccated storage at ambient temperature (Stasis Packs) and 2 species were packaged along with powdered media inside a bioreactor system to allow hydration and growth in microgravity (Production Packs). For both systems, Whole Genome Sequencing (WGS) of the DNA extracted from the returned samples and paired ground controls will be conducted to identify changes in genome stability due to time, storage conditions and growth in space. Across the technical replicates, ground controls, 10 timepoints, and multiple experimental conditions, ~300 samples have been selected for initial analysis with WGS sequencing to 100x coverage. A flexible and resource efficient mutation calling pipeline is needed to process this large dataset and allow for comparisons between species. Many bioinformatics tools for calling Indels and Single Nucleotide Variants (SNVs) are designed for use with pure isolates, where true variations from the reference genome are expected to dominate the reads aligning to the location of mutation. In contrast, DNA from the Stasis Pack (SP) samples was collected directly after recovery from desiccated storage and the Production Pack (PP) samples were collected after fermentation. In this context, reads with mutations are expected to be less frequent than reads that align with the reference genome, as each sample will include multiple lines of cells. Thus, BioNutrients samples are expected to be similar to samples from cancer cell or “pooled” sequencing approaches. In preparation for the analysis of the BioNutrients samples, we have tested three mutation calling tools (GATK for Microbes, BreSeq and DiscoSNP) designed for complex samples. A challenge of validating mutation identification pipelines is a lack of “Ground Truth” datasets, especially for complex samples. To compare these three tools, we sought to identify mutations in pre-existing WGS data collected from populations of Chlamydomonas reinhardtii that were exposed to UV mutagenesis and growth in LEO as part of the Space Algae-1 mission. Here we present a summary of these tools against the analysis originally conducted using the CRISP tool. Critical metrics are compared such as runtime, the number of SNPs, the number and size of Indels, and patterns of transversion and transitions identified by each tool are reported. By sharing these benchmarking results collected in support of the BioNutrients mission, we aim to guide others seeking to identify SNVs in similarly complex microbial samples.

Biology↗