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Does the International Space Station Leak DNA? Preliminary Results from the ISS External Microorganisms Payload

Existing crewed spacecraft like the ISS (International Space Station) leak by design. The ISS routinely releases gas to maintain life support systems and when astronauts exit the station to perform space walks. The chemical component of this leakage is well characterized, but the biological components are not. The ISS is not subject to planetary protection requirements, but planned missions to Mars will use similar systems and will be subject to planetary protection requirements. If detectable microorganisms are escaping through vents and or airlocks we may need to redesign our crewed habitats to minimize this type of contamination. To test the hypothesis that microorganisms from inside ISS are detectable on exterior surfaces an astronaut used the ISS External Microorganisms sampling kit (Rucker et al. 2018) to sample exterior surfaces of the ISS during an EVA (Extra Vehicular Activity) in January of 2025. These samples were returned to Earth for DNA extraction and sequencing. We successfully, extracted and sequenced bacterial, fungal and viral DNA from these samples that was not present in the negative controls. These results should help NASA refine the planetary protection requirements for crewed missions. Methods: The samples were collected using sterile, DNA free, buccal swabs (23 mm. diameter) housed in custom canisters. Each canister uses a 0.2 μm Teflon filter to maintain sterility as the caddy, holding 8 swabs moves in and out of vacuum. The astronaut sampled 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 Carbon Dioxide Removal Assembly vent, and 6) the Vacuum Exhaust System vent. The seventh swab was exposed to vacuum during the EVA without touching it to a surface. The eighth swab, a negative control, was not opened until the caddy returned to Earth. DNA was extracted from the swabs using a QIamp UCP Pathogen kit and prepared for sequencing on an Aviti (Element Biosciences) sequencer (Arslan et al. 2024). The resulting sequences were analyzed using the EDGE Bioinformatics platform (Li et al. 2017). The sequences were analyzed individually using tools like BLAST, GOTTCHA2, Kraken2, and PanGIA. The data were also assembled into metagenome assembled genomes) using tools like CONCOCT, MaxBin2 and MetaBAT2. Results: We successfully extracted and sequenced bacterial, archaeal, fungal and viral DNA from all seven samples. The handrail swab had the lowest number of reads (768,651) and the airlock thermal cover had the highest number of reads (8,819,230). These samples contain DNA from human associated bacteria (e.g. Crynebacterium riegelii ), fungi (.e.g. Penicillium rubens ), and viruses (e.g Alphapapillomavirus ). Conclusion: Preliminary interpretation suggest that the airlock and the space suits themselves are the largest sources of contaminant DNA. Most if not all of the DNA is from organisms known to be present inside the ISS. Vents attached to life support systems may be a lesser source of biological contamination. Further analysis should help NASA address planetary protection knowledge gaps for crewed missions.

Aaron B Regberg

Trajectory Engineering with Modular Patched Conics for Entry Systems and TPS (TEMPEST)

Brief Presenter Biography (35 word limit): Bohdan Wesely is an Aerospace Engineer in the Entry Systems and Technology Division at Ames. He has worked on a variety of projects for NASA including integrated TPS (thermal protection system) flight hardware deliveries and testing services for commercial partners. Introduction: TEMPEST is a new trajectory analysis framework that is designed to fill the gap between dedicated flight mechanics tools and aerothermal and TPS sizing tools. The project started as an SJSU master’s thesis and has since evolved into a general conceptual design tool capable of studying a wide variety of entry problems. Development is ongoing in the Entry Systems and Technology Division at NASA ARC. Why TEMPEST: Space missions involving entry into a planetary atmosphere involve a series of unique requirements across multiple disciplines. Whether it is traditional entry descent and landing (EDL), or aerocapture, the vehicle must navigate to its target landing location or orbit state, and the TPS must protect the payload during entry. The design process typically involves iterative handoffs between various flight mechanics, flow solver, and material response level tools. During the early conceptual phase, a wide variety of feasible trajectories are simulated in a Monte Carlo scenario which broadly satisfy the mission or landing requirements. Next, computational fluid dynamics (CFD), direct simulation Monte Carlo (DSMC), and other flow solver analyses are performed at various key trajectory points to generate an aero-database, heating and TPS design requirements also emerge at this stage. At this point, with updated aerodynamics from the various flow solvers, trajectories can be re-run, this in turn can change the required freestream conditions for the CFD tools, and as a project progresses, these analyses converge, and uncertainty is reduced. However, there is always a “hand-off” occurring between two inherently coupled phenomena. Analysis Description: One of the goals with TEMPEST is to use a variety of first principles estimation methods coupled with an atmosphere model to predict vehicle aerothermodynamics across the entire flight regime while propagating a 3 or 6 degree of freedom (DoF) trajectory. Aerodynamics methods include modified Newtonian, Maxwell and Cercignani- Lampis-Lord (CLL) for continuum, transitional, and free molecular flow regimes. Aerothermodynamics include boundary layer and reference enthalpy methods, and Mutation++ for non-equilibrium chemistry modeling. TEMPEST is also capable of stitching multiple trajectory segments together to study mission scenarios like multi-pass aerocapture and aero-gravity assists. Most of the program is implemented in MATLAB using modern system objects, it relies on several C++ shared libraries for supporting tools like Gmsh, the Global Reference Atmospheric Model (GRAM), and Mutation++. The various first principles aerothermal estimation methods are discretized across either a structured axisymmetric panel mesh or an unstructured tri-mesh generated from an open-source tool such as Gmsh, this allows solutions on the same mesh to be compared across tools such as CB-Aero. CFD Coupling. A physics-aware, gaussian process CFD anchoring scheme is proposed to adjust the various first principles methods as a CFD database is populated. One goal for this anchoring module is to inform the project where CFD should be run. Full knowledge of the entire trajectory, atmosphere, and aerothermodynamics allows for easier identification of high sensitivity areas and uncertainty quantification. While the first principles effects are well known and proven accurate in existing tools such as CB- Aero and Cart3D, a physics aware CFD anchoring scheme increases tool credibility across a project lifecycle. Material Response Modeling. Correct TPS sizing is critical for optimizing mass for science payloads and ensuring mission success. The process typically involves a thermal analysis along the trajectory with surface heating environments as a boundary condition. Several design constraints are maximum bondline temperature and maximum recession with various margining techniques. The material response tool FIAT, developed out of NASA Ames, is currently being integrated into the TEMPEST environment. TPS recession, shape change, mass loss, and mass property alteration are all factors that can perturb an entry trajectory. For missions like Mars 2020, recession was minimal and was safely handled separately as a post process. For missions such as Jupiter Galileo with a high TPS mass fraction or asteroid entries, recession plays a major role. The proposed fully coupled scheme is to use an epoch-based approach where the trajectory integration is halted after a recession threshold, the energy balance and FIAT are solved at each panel, the mesh, aerodynamics, and mass properties are updated, and the trajectory continues. Several computational tradeoffs have been made during the development of TEMPEST to limit the cost of a single trajectory and preserve its utility as a conceptual, rapid iteration tool. Conclusion: Development of TEMPEST is ongoing and the project is still in its infancy. This talk aims to showcase its unique capabilities to support future NASA entry systems missions.

Bohdan O Wesely