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At least 109 records · Page 6

Spaceflight Microbiology

Microbiological Areas of Concern: Astronaut Health; Vehicle integrity; Life support and other systems failure -Biofilm formation/biofouling -Bio-corrosion and biodegradation -Trash and human waste containment -Risk of condensation -Astronaut hygiene areas; Spaceflight foods -Impact of “pick and eat” foods on the environment -Impact of the environment on “pick and eat” foods -Rinsing food with potable water is impractical; Planetary protection -How do we track what we are leaving versus what we are finding in our search for life on other planets?

Ott, C. Mark

Microbiological Analysis of Mizuna Grown in the Veggie Hardware to Define Critical Control Points and Ensure the Safety of Space Grown Crops

The Veggie facility on the International Space Station has been utilized as a “pick and eat” plant growth system to provide fresh produce for crew consumption. The VEG-04 experiments completed in 2019 examined the effect of red-rich and blue-rich light treatments on the growth of mizuna as well as harvest method and resulting yield. Analysis was performed on plant tissues and associated hardware to evaluate the influence of these experimental variables on the microbial population. VEG-04A plant pillows with pre-planted Mizuna mustard seeds were launched on SpaceX-16 in December 2018. The pillows were initiated, and a single 35-day harvest was performed. Veg 04B pillows were pre-planted with Mizuna seeds and launched on SpaceX-18 in July 2019, initiated and subsequently harvested at days 29, 43 and 58. The crew consumed approximately half of the produce, and the remainder was frozen and returned for analysis. Leaves, swabs, wicking material, substrate, and roots were processed and plated on media for the enumeration and isolation of bacteria and fungi. Isolated bacterial colonies were identified using Biolog Micro ID system or MicroSEQ16S rDNA sequencing technique. Fungal colonies were identified using the MicroSEQ D2 rDNA kit. Sample extracts were plated onto specialized media to identify Escherichia coli/coliforms, Staphylococcus aureus and Salmonella sp. Results indicate that bacterial and fungal counts were higher in plants grown in red-rich lightin VEG-04A, while the opposite was true in the Veg-04B third harvest. Microbial counts increased with the repeated harvest method used in Veg-04B. These data support the understanding of environmental and horticultural practices that can affect the microbiological quality of space-grown produce grown and aid in identification of critical control points for the development of a hazard analysis critical control point plan for ISS-grown crops. This research was co-funded by the NASA’s Human Research Program and Space Biology.

Mary E Hummerick

The Microbiology of Microgreens Grown in Controlled Environment Chambers under ISS Conditions

Microgreens have been identified as a new type of pick-and-eat salad crop that can be utilized in space crop production systems. The majority of traditionally grown leafy green crops can be grown as microgreens, in addition to crops such as legumes, sunflower, buckwheat, most herbs, and corn, presenting hundreds of microgreen crop options. Notably, microgreens are nutrient dense, high in beneficial compounds like antioxidants, Vitamins C and K, and exhibit a variety of desirable flavors and textures.The short growth cycles (7-14 days), low water requirements and volume optimization potential make them a viable option for sustainable production of nutritious and flavorful crops in space.The crop production team at Kennedy Space Center is investigating the food safety aspects of microgreens grown under spaceflight relevant conditions for crew consumption. Microbiological analysis and screening for potential foodborne pathogens was performed on over 20 varieties of microgreens that have demonstrated positive horticultural attributes.Additionally, a comparison of microgreens grown hydroponically under ISS environmental conditions and similar varieties from local markets was completed to collect baseline data on the microbial load on microgreens. In aneffort to improve microgreen quality, strategies to reduce the microbial load were tested, including bulk seed sanitization, harvest age, exposure to high blue light, and post-harvest chemical disinfection. The efficacy of a citric acid-based produce wash currently used for ISS grown produce and 1% H2O2were investigated at different exposure times for reduction in bacterial and fungal counts on a variety of microgreens. Limited log reduction was achieved depending on exposure time.Our testing also demonstrated that seed sanitization impacted microbial load on microgreens and systems.

Mary E. Hummerick

Intrabasin Comparison of the Microbiology and Geochemistry of Produced Fluid From Hydraulically Fractured Wells in the Permian Region

The Permian Basin is the highest producing oil reservoir in the United States. Hydrocarbon extraction methods in this region are often associated with frac hits, or interwell communication events where an established well is affected by the pumping of fracture fluid into a new well. Our previous work revealed a geochemical signal indicating the presence of frac hits in the Permian Basin. We returned to this area with the goal of expanding our understanding of subsurface interactions common in this region. To do so, we collected produced water from 25 unique sites across the Permian Basin, 10 of which had previously been characterized during an active frac hit. For each sample, we measured the pH, alkalinity, geochemistry, microbial load, and microbial community composition. Permian Basin produced water is characterized by higher sulfate and lower total dissolved solids (TDS) concentrations compared to other regions. Interestingly, wells impacted by frac hits have a geochemical profile that resembles that of fracture fluid, with both lowered sulfate and lowered TDS concentrations compared to unaffected wells. Due to the year-long recovery window between sample collection periods, we anticipate that all our data will be characterized by the typical high sulfate, low TDS concentrations.

geochemistry

Post-Viking microbiology: new approaches, new data, new insights

In the 20 years since the Viking experiments, major advances have been made in the areas of microbial systematics, microbial metabolism, microbial survival capacity, and the definition of environments on earth, suggesting that life is more versatile and tenacious than was previously appreciated. Almost all niches on earth which have available energy, and which are compatible with the chemistry of carbon-carbon bonds, are known to be inhabited by bacteria. The oldest known bacteria on earth apparently evolved soon after the formation of the planet, and are heat loving, hydrogen and/or sulfur metabolizing forms. Among the two microbial domains (kingdoms) is a great deal of metabolic diversity, with members of these forms being able to grow on almost any known energy source, organic or inorganic, and to utilize an impressive array of electron acceptors for anaerobic respiration. Both hydrothermal environments and the deep subsurface environments have been shown to support large populations of bacteria, growing on energy supplied by geothermal energy, thus isolating these ecosystems from the rest of the global biogeochemical cycles. This knowledge, coupled with new insights into the history of the solar system, allow one to speculate on possible evolution and survival of life forms on Mars.

Review

Ecosystems and Networks Integrated with Genes and Molecular Assemblies (ENIGMA): Molecular and Computational Technologies for Environmental Microbiology (Final Scientific/Technical Report)

The ENIGMA science focus area (SFA) is a multi-disciplinary, multi-institutional research effort focused on addressing foundational knowledge gaps in environmental microbial communities by studying groundwater and sediment microbiomes in the shallow subsurface at the contaminated Oak Ridge Reservation (ORR). We seek to discover and characterize the reciprocal interactions between the microbial communities and the geochemical and geophysical parameters of the shallow subsurface within the contamination plume. The primary goal of this subcontract was to develop experimental and computational tools to advance our understanding of microbial adaptation and community assembly in contaminated environments, with specific efforts in high-throughput genomic methods, microbial ecology tools, and studies of heavy metal contamination impacts.

54 ENVIRONMENTAL SCIENCES

Rapid Design and Engineering of Smart and Secure Microbiological Systems (Final Report)

The design and application of successfully engineered biosystems requires an understanding of how engineered microbes will interact with other organisms – either as one-on-one competitors or in the context of microbial consortia. Engineering microorganisms from first principles for non-laboratory, environmental applications is inherently challenging because: (1) engineered systems tend to quickly revert back to their wild-type behaviors; and (2) these systems typically pay a price in reduced fitness, making them uncompetitive against invasive contaminating species (i.e., metabolic burden). For this project, we used a synthetic biology-based strategy to investigate the organization, control, stabilization, and destabilization of natural and engineered microbes. This approach enabled development of (1) single-strain systems capable of detecting and responding to target organisms in the environment; (2) a pipeline for refining and engineering biological constructs in new, non-model host organisms; and (3) improved systems for rapidly designing, engineering, and assaying new biological modules. This coupled approach to safeguard system design is predictable and portable across bacterial species and is focused on microbes that are part of the beneficial plant microbiome. A long-term goal beyond the proposed research is to enable the rational engineering of microbial communities based on first principles of biological design that mimic the smart performance of microorganisms observed in natural systems.

59 BASIC BIOLOGICAL SCIENCES

Characterization of Coal Seam Produced Water from the Appalachian Basin: Geochemistry, Microbiology, and Biocatalyst Development for Biological CO 2 Conversion

To enhance domestic energy repositories, it is necessary to develop carbon utilization technologies to repurpose unused carbon sources, and/or enhance the economic viability and carbon conversion efficiency of current technologies. To this end we have developed a biocatalyst, enriched from coal seam produced water, to convert gaseous and liquid C1 compounds (e.g. CO 2 , bicarbonate, formate, and/or methanol) into acetate, a C2 organic acid with a projected global market size of $\$$32.4 billion USD by 2030.

01 COAL, LIGNITE, AND PEAT