Isolation and Monitoring of Cleanroom-Associated Microbial Contaminates From Geological Collections

Microbial contamination is of particular interest to geological curation as many microorganisms can change mineral composition and produce compounds used as biosignatures used for the detection of life. Microbial cells can change the mineral composition of rocks through organic acid production and direct enzymatic oxidation/reduction of transition metals. Enzymatic oxidation of iron and manganese can occur at a rate several orders of magnitude faster than under abiotic conditions and produce highly reactive nanoparticle- sized oxides that can react and sorb other metals and organic compounds. Many fungi can also produce organic acids that dissolve and chelate mineral matrices chemically reducing and dissolving rock surfaces. Finally, several common soil-associated bacteria and fungi produce secondary metabolites that contain unusual amino acid analogs and non-ribosomal peptides containing both L- and D- chirality used in characterizing carbonaceous chondrites and the detection of extraterrestrial life

Mobile/Modular BSL-4 Containment Facilities Integrated into a Curation Receiving Laboratory for Restricted Earth Return Missions

NASA robotic sample return missions designated Category V Restricted Earth Return by the NASA Planetary Protection (PP) Office require sample containment and biohazard testing upon return to Earth. Since the 1960s, sample containment from an unknown extraterrestrial biohazard have been related to the highest containment standards and protocols known to modern science. Today, this is Biosafety Level (BSL) 4 containment. In the U.S., the Biosafety in Microbiological and Biomedical Laboratories publication authored by the U.S. Department of Health and Human Services (HHS): Public Health Service, Centers for Disease Control and Prevention, and the National Institutes of Health houses the primary recommendations, standards, and design requirements for all BSL labs. Past mission concept studies for constructing a NASA Curation Receiving Laboratory with an integrated BSL-4 quarantine and biohazard testing facility have been estimated in the hundreds of millions of dollars (USD). As an alternative option, we have conducted a trade study for constructing a mobile and/or modular sample containment laboratory that would meet all BSL-4 and planetary protection standards and protocols at a fraction of the cost. Mobile and modular BSL-2 and 3 facilities have been successfully constructed and deployed world-wide for government testing of pathogens and pharmaceutical production. Our study showed that a modular BSL-4 construction could result in ~ 90% cost reduction when compared to traditional BSL-4 construction methods without compromising the preservation of the samples or Earth. For the design/construction requirements of a mobile/modular BSL-4 containment, we used the established HHS document standards and protocols for manipulation of agents in Class III Biosafety Cabinets (BSC; i.e., negative pressure gloveboxes) that are currently followed in operational BSL-4 facilities in the U.S

Organic Biomarker-Based Assays to Evaluate Total Bioburden and Organic Compounds on Space Flight Hardware

Meeting planetary protection (PP) requirements for space flight hardware may involve bioburden reduction by dry heat microbial reduction (DHMR). The NASA standard assay to demonstrate the reduction of organisms involves the swabbing of surfaces, heat shock of the extracted samples, plating of the samples on Trypticase Soy Agar (TSA), and counting colony forming units after an incubation period. The standard assay uses enumeration of heat tolerant spore-formers as a proxy for total bioburden and is generally expected to provide a lower limit. We suggest that a better estimate of the total bioburden could be obtained through sampling and analysis of organic biomarkers. As biological organisms are fundamentally organic in chemistry (i.e. carbon containing materials) it is important to characterize the biomarker compounds that are released from organisms that 1) exist on flight hardware before microbial reduction and 2) left behind from the killed organisms following microbial reduction

Curating Nasa's Future Extraterrestrial Sample Collections: the Role of Advanced Curation

The Astromaterials Acquisition and Curation Office at NASA Johnson Space Center (JSC) (henceforth referred to herein as NASA Curation Office) is responsible for curating all of NASA's extraterrestrial samples. Under the governing document, NASA Policy Directive (NPD) 7100.10F "Curation of Extraterrestrial Materials," JSC is charged with "The curation of all extraterrestrial material under NASA control, including future NASA missions." The Directive goes on to define Curation as including "...documentation, preservation, preparation, and distribution of samples for re-search, education, and public outreach." Here we describe some of the ongoing efforts to ensure that the future activities of the NASA Curation Office are working towards a state of maximum proficiency

In-situ Optimized Substrate Witness Plates: Ground Truth for Key Processes on the Moon and Other Planets

Future exploration efforts of the Moon, Mars and other bodies are poised to focus heavily on persistent and sustainable survey and research efforts, especially given the recent interest in a long-term sustainable human presence at the Moon. Key to these efforts is understanding a number of important processes on the lunar surface for both scientific and operational purposes. We discuss the potential value of in-situ artificial substrate witness plates, powerful tools that can supplement familiar remote sensing and sample acquisition techniques and provide a sustainable way of monitoring processes in key locations on planetary surfaces while maintaining a low environmental footprint. These tools, which we call Biscuits, can use customized materials as wide ranging as zircon-based spray coatings to metals potentially usable for surface structures, to target specific processes/questions as part of a small, passive witness plate that can be flexibly placed with respect to location and total time duration. We examine and discuss unique case studies to show how processes such as water presence/transport, presence and contamination of biologically relevant molecules, solar activity related effects, and other processes can be measured using Biscuits. Biscuits can yield key location sensitive, time integrated measurements on these processes to inform scientific understanding of the Moon and enable operational goals in lunar exploration. While we specifically demonstrate this on a simulated traverse and for selected examples, we stress all groups interested in planetary surfaces should consider these adaptable, low footprint and highly informative tools for future exploration.Comment: Accepted to Earth and Space Science, Will be updated upon publicatio

Preliminary Planning for Mars Sample Return (MSR) Curation Activities in a Sample Receiving Facility

The Mars Sample Return Planning Group 2 (MSPG2) was tasked with identifying the steps that encompass all the curation activities that would happen within the MSR Sample Receiving Facility (SRF) and any anticipated curation-related requirements. An area of specific interest is the necessary analytical instrumentation. The SRF would be a Biosafety Level-4 facility where the returned MSR flight hardware would be opened, the sample tubes accessed, and the martian sample material extracted from the tubes. Characterization of the essential attributes of each sample would be required to provide enough information to prepare a sample catalog used in guiding the preparation of sample-related proposals by the world’s research community and informing decisions by the sample allocation committee. The sample catalog would be populated with data and information generated during all phases of activity, including data derived concurrent with Mars 2020 sample-collecting rover activity, sample transport to Earth, and initial sample characterization within the SRF. We conclude that initial sample characterization can best be planned as a set of three sequential phases, which we have called Pre-Basic Characterization (Pre-BC), Basic Characterization (BC), and Preliminary Examination (PE), each of which requires a certain amount of instrumentation. Data on specific samples and subsamples obtained during sample safety assessments and time-sensitive scientific investigations would also be added to the catalog. There are several areas where future work would be beneficial to prepare for the receipt of samples, which would include the design of a sample tube isolation chamber and a strategy for opening the sample tubes and removing dust from the tube exteriors

Planning Implications Related to Sterilization-Sensitive Science Investigations Associated with Mars Sample Return (MSR)

The NASA/ESA Mars Sample Return (MSR) Campaign seeks to establish whether life on Mars existed where and when environmental conditions allowed. Laboratory measurements on the returned samples are useful if what is measured is evidence of phenomena on Mars rather than of the effects of sterilization conditions. This report establishes that there are categories of measurements that can be fruitful despite sample sterilization and other categories that cannot. Sterilization kills living microorganisms and inactivates complex biological structures by breaking chemical bonds. Sterilization has similar effects on chemical bonds in non-biological compounds, including abiotic or pre-biotic reduced carbon compounds, hydrous minerals, and hydrous amorphous solids. We considered the sterilization effects of applying dry heat under two specific temperature-time regimes and the effects of γ-irradiation. Many measurements of volatile-rich materials are sterilization sensitive—they will be compromised by either dehydration or radiolysis upon sterilization. Dry-heat sterilization and γ-irradiation differ somewhat in their effects but affect the same chemical elements. Sterilization-sensitive measurements include the abundances and oxidation-reduction (redox) states of redox-sensitive elements, and isotope abundances and ratios of most of them. All organic molecules, and most minerals and naturally occurring amorphous materials that formed under habitable conditions, contain at least one redox-sensitive element. Thus, sterilization-sensitive evidence about ancient life on Mars and its relationship to its ancient environment will be severely compromised if the samples collected by Mars 2020 rover Perseverance cannot be analyzed in an unsterilized condition. To ensure that sterilization-sensitive measurements can be made even on samples deemed unsafe for unsterilized release from containment, contingency instruments in addition to those required for curation, time-sensitive science, and the Sample Safety Assessment Protocol would need to be added to the Sample Receiving Facility (SRF). Targeted investigations using analogs of MSR Campaign-relevant returned-sample types should be undertaken to fill knowledge gaps about sterilization effects on important scientific measurements, especially if the sterilization regimens eventually chosen are different from those considered in this report

Science and Curation Considerations for the Design of a Mars Sample Return (MSR) Sample Receiving Facility

The most important single element of the “ground system” portion of a Mars Sample Return (MSR) Campaign is a facility referred to as the Sample Receiving Facility (SRF), which would need to be designed and equipped to receive the returned spacecraft, extract and open the sealed sample container, extract the samples from the sample tubes, and implement a set of evaluations and analyses of the samples. One of the main findings of the first MSR Sample Planning Group (MSPG, 2019a) states that “The scientific community, for reasons of scientific quality, cost, and timeliness, strongly prefers that as many sample-related investigations as possible be performed in PI-led laboratories outside containment.” There are many scientific and technical reasons for this preference, including the ability to utilize advanced and customized instrumentation that may be difficult to reproduce inside in a biocontained facility, and the ability to allow multiple science investigators in different labs to perform similar or complementary analyses to confirm the reproducibility and accuracy of results. It is also reasonable to assume that there will be a desire for the SRF to be as efficient and economical as possible, while still enabling the objectives of MSR to be achieved. For these reasons, MSPG concluded, and MSPG2 agrees, that the SRF should be designed to accommodate only those analytical activities that could not reasonably be done in outside laboratories because they are time- or sterilization-sensitive, are necessary for the Sample Safety Assessment Protocol (SSAP), or are necessary parts of the initial sample characterization process that would allow subsamples to be effectively allocated for investigation. All of this must be accommodated in an SRF, while preserving the scientific value of the samples through maintenance of strict environmental and contamination control standards