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At least 73 records · Page 4

Spaceflight-Induced Changes in Microbial Virulence and the Impact to the Host Immune Response

Many microbial pathogen shave repeatedly exhibited unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in final cell concentration, biofilm production, stress resistance, antibiotic sensitivity, gene expression, host-pathogen interactions, and virulence. Notably, the classic foodborne pathogen Salmonella enterica serovar Typhimurium displayed increased virulence in animals when cultured in either the spaceflight analogue or true spaceflight environment. Recently, Serratia marcescens also was shown to increase virulence when cultured in the spaceflight environment. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system at multiple levels, which suggests an increased risk of infectious diseases. Moreover, astronauts have some degree of clinical infectious disease incidence. However, the contribution of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. The goal of this study is to gain insight into the breadth of other medically significant microbial pathogens that may exhibit altered virulence and pathogenesis-related responses when cultured in space flight analogue conditions. Specifically, we are characterizing the effect of spaceflight analogue culture (Low Shear Modeled Microgravity/LSMMG) on microbial pathogenesis-related stress responses, in vitro host-pathogen interactions, gene expression, and virulence potential in animals using five important model bacterial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli. Herein, we present data from one of these pathogens, the foodborne bacterium, S. enterica Enteritidis, which is closely related to S. enterica Typhimurium. Phenotypes evaluated included growth profiles, environmental stress responses(acid, oxidative, bile, and thermal stresses),and in vitro colonization of3-D biomimetic cultures of human intestinal tissue containing immune cells. Transcriptomic profiling and virulence studies are ongoing. We show that S. Enteritidis exhibited key alterations in pathogenic responses to LSMMG culture that suggest increased infection risk, including several responses which were different from those observed in the closely related pathovar S. Typhimurium. This information will provide critical mechanistic insight into the potential impact of microgravity on alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.

C.M. Ott↗

Tissuelike 3D Assemblies of Human Broncho-Epithelial Cells

Three-dimensional (3D) tissuelike assemblies (TLAs) of human broncho-epithelial (HBE) cells have been developed for use in in vitro research on infection of humans by respiratory viruses. The 2D monolayer HBE cell cultures heretofore used in such research lack the complex cell structures and interactions characteristic of in vivo tissues and, consequently, do not adequately emulate the infection dynamics of in-vivo microbial adhesion and invasion. In contrast, the 3D HBE TLAs are characterized by more-realistic reproductions of the geometrical and functional complexity, differentiation of cells, cell-to-cell interactions, and cell-to-matrix interactions characteristic of human respiratory epithelia. Hence, the 3D HBE TLAs are expected to make it possible to perform at least some of the research in vitro under more-realistic conditions, without need to infect human subjects. The TLAs are grown on collagen-coated cyclodextran microbeads under controlled conditions in a nutrient liquid in the simulated microgravitational environment of a bioreactor of the rotating- wall-vessel type. Primary human mesenchymal bronchial-tracheal cells are used as a foundation matrix, while adult human bronchial epithelial immortalized cells are used as the overlying component. The beads become coated with cells, and cells on adjacent beads coalesce into 3D masses. The resulting TLAs have been found to share significant characteristics with in vivo human respiratory epithelia including polarization, tight junctions, desmosomes, and microvilli. The differentiation of the cells in these TLAs into tissues functionally similar to in vivo tissues is confirmed by the presence of compounds, including villin, keratins, and specific lung epithelium marker compounds, and by the production of tissue mucin. In a series of initial infection tests, TLA cultures were inoculated with human respiratory syncytial viruses and parainfluenza type 3 viruses. Infection was confirmed by photomicrographs that showed signs of damage by viruses and virus titers (see figure) that indicated large increases in the populations of viruses during the days following inoculation.

Goodwin, Thomas J.↗

Responses of Microbes to Modeled Space Radiation

The built environment of spaceships is host to a microbial community that affects crew and craft alike. While the static composition of this community has been characterized and its temporal dynamics examined, the mechanisms controlling its make-up and evolutionary trajectory are not understood. Systematic analyses of microbial diversity show consistent patterns in community composition and function. Understanding these patterns' ecological origins remains a significant challenge, as it requires connecting processes at varying temporal and spatial scales. However, it is clear that the state and trajectories of microbial communities are in-part determined by their physical environment. In this regard, the spaceflight environment includes numerous interacting factors that differentiate it from Earth environments, including an altered atmospheric composition, reduced gravity (and thus altered fluid dynamics), and increased ionizing radiation. These factors impart selective pressures on microbial communities that affect their evolutionary trajectories and thus the risks and benefits these communities represent to crew and craft. The radiation environment of space leads to chronic exposure to low doses and is difficult to mimic on Earth. Thus, little is known about how microbial communities in spacecraft will respond and evolve. Therefore, given the limitations of existing studies, we aim to empirically determine how exposure to low doses of ionizing radiation for thousands of cell divisions affects rates of mutation accumulation in bacteria and the trajectory of their evolution. In this way, we will provide a critical set of data for designing safe and robust space missions. Here we discuss our progress towards this aim, including the construction of exposure facilities, our culturing and analysis approach, and preliminary data.

radiation↗

VEG-04: the Effects of Light Quality on Mizuna Mustard Growth, Nutritional Composition, and Organoleptic Acceptability for a Space Diet

Space crop production will be important in future long duration exploration missions to supplement the packaged diet with fresh bioactive nutrients. Plant care and the addition of fresh veggies to the diet may also have a role in astronaut well-being. Pick-and-eat salad crops are the best candidates for this near-term supplementation since they require minimal processing or preparation to add to meals. While light quality can strongly influence plant responses on Earth, the impacts of light quality on plant growth and composition in spaceflight remain unclear. The VEG-04 experiment uses two Veggie plant growth chambers on the International Space Station to simultaneously test different red: blue light ratios on the growth of Mizuna mustard, a leafy green salad crop. In addition to plant health and yield, the composition of key nutrients is assessed. Astronauts conduct on-board organoleptic evaluation of the fresh produce. Microbial food safety of returned produce is examined, and a Hazard Analysis Critical Control Point (HACCP) plan has been developed for this crop. VEG-04 consists of two experiments, one lasting 28 days with a single harvest, and the second lasting 56 days, with three cut-and-come-again harvests. These different scenarios provide an opportunity to test two production concepts, examine different fertilizers, monitor microbial changes over time for this crop, and assess potential impacts of interacting with plants on crew behavioral health and performance in spaceflight operations. In ground testing, plant growth was not significantly different across the different light treatments, however nutrient composition did differ significantly. Flight test results will be compared with ground data. This research was co-funded by NASA's Human Research Program and Space Biology in the ILSRA 2015 NRA call.

Massa, Gioia D.↗

EnviroNET

EnviroNET is a service/facility that provides users with on-line, dial-up technical information concerning environmental conditions likely to be encountered by instruments and experimental arrangements carried aboard spacecraft. EnviroNET incorporates at present a combination of expository text and numerical tables amounting to about two million characters (bytes), plus FORTRAN programs that model the space environment. The topics covered are presented in viewgraph form and include the following: advantages of EnviroNET; thermal and humidity; vibration and acoustics; electromagnetic interference; loads and low frequency dynamics; microbial and toxic contaminants; molecular contamination; natural environment; orbiter motion; particulate environment; surface interactions; spacecraft anomalies; and interactive graphics facility.

Vansant, Tim↗

Squeezing Every Last 'Bit' of Information from Enceladus Mass Spectrometry

Potential opportunities to return to Enceladus in Discovery and Flagship class missions inspire development of next-generation instruments and creative approaches to sample collection, sample analysis, and data analysis and transmission strategies. Mass spectrometers (MS) are ideally suited to future Enceladus missions due to their analytical power in identifying a range of molecular and ionic compositions – including complex organics – and potentially astrobiologically-important features such as isotope ratios, chirality, and enantiomeric excess. However, long communication delays from Enceladus and limited bandwidth limits the data transmission from these higher-data-volume instruments, likely delaying mission-related response to new data. We explore the utility of data science and machine learning (ML) on isotope ratio (IR)MS data collected from laboratory analogs of Enceladus to: 1) process data quickly for rapid ground-based analyses, 2) understand if compositional and biosignature information could be extracted from IRMS data, and 3) evaluate whether onboard ML techniques could improve sample analysis, cadence, and transmission prioritization. Laboratory analogs analyzed isotopes of volatile CO2 that interacted with seawaters of varying composition, and include both abiotic and biotic (microbially-influenced) experiments. Enceladus’s alkaline oceans promote speciation of carbon into multiple forms (e.g., H2CO3 / CO2, HCO3-, and CO32-), each of which could be isotopically fractionated by abiotic or biotic reactions. Large (>2‰) changes in carbon isotopes (δ13C) are observed from some biotic experiments inoculated with complex microbial ecosystems relative to the abiotic seawaters. ML training and classification suggests that microbial samples can be distinguished from abiotic samples, yet that a broad range of microbial experiments are necessary to train ML models to cover a range of complexities including disequilibria, and isotopic and compositional fractionation.

geochemistry↗

Automating the Study of Microbial Adaptation Dynamics on and off the ISS

The International Space Station (ISS) not only serves as a unique environment for humans, but also the microorganisms that join alongside. Many microbes present on the spacecraft arrive via humans, and as they interact with different surfaces they begin to inhabit those locations. Much like how human health has shown to be impacted by these extreme environments, microbial viability and response to stress also changes. Experimental evolution (EE) can aid in studying how microbes’ growth and activity changes within the ISS environments by applying controlled stressors to microbial cultures and monitoring their response over generations. EE studies are commonly done manually in laboratories, but, with multiple environmental variables to measure and adjust, it becomes highly labor-intensive, prone to human error, and challenging to scale. A multipurpose automated EE system named the AADEC has been developed to address these problems. This system integrates multiple sensors into a single fluidic chamber using UV-C flux, temperature, and media composition as stressors. AADEC contains five sensors: oxidation-reduction potential, electrical conductivity, pH, dissolved oxygen, and optical density. On their own, each is able to provide certain information on growth rate or metabolism; together, they show in detail how stressors affect life. AADEC studies can be conducted on Earth and repeated aboard the ISS to see how behavior changes when exposed to space mission stressors such as microgravity and radiation. AADEC’s auxiliary systems include peristaltic pumps for media exchange, magnetic rods for agitation, and a Raspberry Pi microprocessor to monitor, store, and adjust stressor levels real-time. This allows researchers to gather information within rapid generations, data and accuracy which is challenging to achieve through manual studies. With further miniaturization and automation, such as a more robust single-piece fluidics card, AADEC has the potential to be developed as a spacecraft payload. Support: NASA Ames CIF Award

Automating↗

Astrobiology, Mars Exploration and Lassen Volcanic National Park

The search for evidence of life beyond Earth illustrates how the charters of NASA and the National Park Service share common ground. The mission of NPS is to preserve unimpaired the natural and cultural resources of the National Park System for the enjoyment, education and inspiration of this and future generations. NASA's Astrobiology program seeks to understand the origins, evolution and distribution of life in the universe, and it abides by the principles of planetary stewardship, public outreach, and education. We cannot subject planetary exploration destinations to Earthly biological contamination both for ethical reasons and to preserve their scientific value for astrobiology. We respond to the public's interest in the mysteries of life and the cosmos by honoring their desire to participate in the process of discovery. We involve youth in order to motivate career choices in science and technology and to perpetuate space exploration. The search for evidence of past life on Mars illustrates how the missions of NASA and NPS can become synergistic. Volcanic activity occurs on all rocky planets in our Solar System and beyond, and it frequently interacts with water to create hydrothermal systems. On Earth these systems are oases for microbial life. The Mars Exploration Rover Spirit has found evidence of extinct hydrothermal system in Gusev crater, Mars. Lassen Volcanic National Park provides a pristine laboratory for investigating how microorganisms can both thrive and leave evidence of their former presence in hydrothermal systems. NASA scientists, NPS interpretation personnel and teachers can collaborate on field-oriented programs that enhance Mars mission planning, engage students and the public in science and technology, and emphasize the ethics of responsible exploration.

Lassen Volcanic National Park↗

Astrobiology Investigations at a Martian Hematite Site

Christensen et al, using data from the Mars Global Surveyor Thermal Emission Spectrometer (TES), have identified gray crystalline hematite in a 350 km by 750 km region near Sinus Meridiani. The deposit corresponds closely to the low-albedo highlands unit 'sm', mapped as a wind-eroded, ancient, subaqueous sedimentary deposit. Christensen et al interpreted the Sinus Meridiani deposit to be 'an in-place, rock-stratigraphic sedimentary unit characterized by smooth, friable layers composed primarily of basaltic sediments with approximately 10 to 15 % crystalline gray hematite.' Christensen et al discussed five possible mechanisms for the formation of this deposit: direct precipitation from standing, oxygenated, Fe-rich water; precipitation from Fe-rich hydrothermal fluids; low-temperature dissolution and precipitation through mobile groundwater leaching; surface weathering and coatings; thermal oxidation of magnetite-rich lavas. Four of these mechanisms involve the interactions of rock with water, and thus have implications in the search for evidence of microbial life.

Allen, Carlton, C.↗

Meridiani Planum Hematite Deposit: Potential for Preservation of Microfossils

Christensen et al., using data from the Mars Global Surveyor Thermal Emission Spectrometer (TES), have identified gray crystalline hematite in a 350 km by 750 km region near Meridiani Planum. The deposit corresponds closely to the low-albedo highlands unit sm, mapped as a wind-eroded, ancient, subaqueous sedimentary deposit. Christensen et al. interpreted the Meridiani Planum deposit to be an in-place, rock-stratigraphic sedimentary unit characterized by smooth, friable layers composed primarily of basaltic sediments with approximately 10 to 15% crystalline gray hematite. The Meridiani Planum hematite deposit has recently been designated as the prime landing site for one of the two Mars Exploration Rover (MER) spacecraft. The MER landings are scheduled for January, 2004. Christensen et al. discussed five possible mechanisms for the formation of this deposit: direct precipitation from standing, oxygenated, Fe-rich water; precipitation from Fe-rich hydrothermal fluids; low-temperature dissolution and precipitation through mobile groundwater leaching; surface weathering and coatings; thermal oxidation of magnetite-rich lavas. Four of these mechanisms involve the interactions of rock with water, and thus have implications in the search for evidence of microbial life.

C C Allen↗

Mitigation of Silver Ion Loss from Solution by Polymer Coating of Metal Surfaces, Part IV

Ionic silver (Ag+) biocide is a leading candidate to provide residual microbial control in spacecraft potable water systems, but suffers from rapid concentration loss due to interactions with the metallic storage containers and tubing. One approach to resolve this problem is the coating of metal surfaces with an inert barrier film. In previous reports, we have described our investigations addressing Ag+ loss mitigation and adhesion performance of parylene barrier coatings on coupons of several metal alloys and 316Ltubing under static immersion. In such conditions, parylene-C and -AF4 coatings have shown excellent Ag+loss mitigation and mixed long-term adhesion performance, depending on parylene species and substrate surface chemistry/structure. In Part IV of this series, we report on our work to investigate the performance of parylene-C barrier coatings, under more challenging and realistic conditions, in order to evaluate potential suitability for use. The resilience and associated Ag+ loss with Parylene-C coating on½” (12.7 mm)316L tubing and fittings under medium-term immersion and repeated fitting dis/reassembly were investigated. Potential mechanical challenges to barrier coatings in spacecraft potable water systems were investigated, with two main focuses being identified: liquid flow/pressure cycling in tubing and cyclic operation of bellows in positive-expulsion storage tanks. Two corresponding testbeds for experimental characterization of coating performance were developed. In the Flow/Pressure Testbed System, Ag+ solution will be pumped through internally coated 316L tubing, with optional pressure cycling. In the Bellows Tank Testbed System, an internally coated edge-welded 316L bellows containing Ag+ solution will be cyclically extended and compressed, analogous to the operation of bellows tanks used for potable water storage and delivery on the International Space Station. The design and operation of these testbeds are described. Finally, the Ag+ adsorption and saturation behaviors of Kalrez and Viton, for potential use as seal materials in fittings and other connections, were characterized in limited experiments.

mitigation↗

Marine Microbial Mats and the Search for Evidence of Life in Deep Time and Space

Cyanobacterial mats in extensive seawater evaporation ponds at Guerrero Negro, Baja California, Mexico, have been excellent subjects for microbial ecology research. The studies reviewed here have documented the steep and rapidly changing environmental gradients experienced by mat microorganisms and the very high rates of biogeochemical processes that they maintained. Recent genetic studies have revealed an enormous diversity of bacteria as well as the spatial distribution of Bacteria, Archaea and Eukarya. These findings, together with emerging insights into the intimate interactions between these diverse populations, have contributed substantially to our understanding of the origins, environmental impacts, and biosignatures of photosynthetic microbial mats. The biosignatures (preservable cells, sedimentary fabrics, organic compounds, minerals, stable isotope patterns, etc.) potentially can serve as indicators of past life on early Earth. They also can inform our search for evidence of any life on Mars. Mars exploration has revealed evidence of evaporite deposits and thermal spring deposits; similar deposits on Earth once hosted ancient microbial mat ecosystems.

Des Marais, David J.↗

Deep Space Gateway Science Opportunities

The NASA Life Sciences Research Capabilities Team (LSRCT) has been discussing deep space research needs for the last two years. NASA's programs conducting life sciences studies - the Human Research Program, Space Biology, Astrobiology, and Planetary Protection - see the Deep Space Gateway (DSG) as affording enormous opportunities to investigate biological organisms in a unique environment that cannot be replicated in Earth-based laboratories or on Low Earth Orbit science platforms. These investigations may provide in many cases the definitive answers to risks associated with exploration and living outside Earth's protective magnetic field. Unlike Low Earth Orbit or terrestrial locations, the Gateway location will be subjected to the true deep space spectrum and influence of both galactic cosmic and solar particle radiation and thus presents an opportunity to investigate their long-term exposure effects. The question of how a community of biological organisms change over time within the harsh environment of space flight outside of the magnetic field protection can be investigated. The biological response to the absence of Earth's geomagnetic field can be studied for the first time. Will organisms change in new and unique ways under these new conditions? This may be specifically true on investigations of microbial communities. The Gateway provides a platform for microbiology experiments both inside, to improve understanding of interactions between microbes and human habitats, and outside, to improve understanding of microbe-hardware interactions exposed to the space environment.

Quincy, C. D.↗

Planetary Evolution and Microbe-Environment Interactions

During the three-year period of this Cooperative Agreement my laboratory conducted physiological and ecological studies regarding the abiotic/biotic relationship in microbial ecosystems. The overall objective of the research performed in the three years of this cooperative agreement is to determine how living systems respond to environmental stress. To accomplish this objective microbes were subjected to a variety of different environmental stresses including decreases in water activity, changes in radiation flux, and nutrient limitation. This research addressees two subsets of the astrobiology questions including: If life arose on early Mars what would have become of it once Mars lost most of its atmosphere and water? and if Earth organisms traveled to Mars what would become of them. To help answer this question we used microbes as model systems not only because they were the first living systems to arise, but also because they are the most likely to be able to withstand a broad range of environmental factors. To meet this objective we conducted laboratory and field work.

Mancinelli, Rocco L.↗

Nearest-Neighbor Machine Learning Feature Selection for Interpretation of Microbial Molecular Signatures from Isotope Ratio Mass Spectrometry Data

Mass spectrometry (MS) promises to be a powerful tool for potential biosignature detection during astrobiological missions on ocean worlds in our solar system. Accurate and generalizable machine learning methods could enhance science return on investment by predicting seawater chemistry and classifying isotopic biosignatures, either as a signature consistent with microbial life (biotic) or as a novelty (unclassified/unique). However, machine learning models are likely to be complex and involve interactions between MS features, making biosignatures difficult to interpret. Feature selection methods provide biological and chemical context that help interpret the mechanisms of machine learning models, but these methods also need the ability to detect complex interactions. Previously, we developed a machine learning feature selection algorithm called nearest-neighbor projected distance regression (NPDR) that has the ability to identify important model features that involve complex interactions and automatically reduce correlation and the dimensionality in a high-dimensional variable space. The standard distance metrics used in NPDR – Manhattan and Euclidean – assume the multivariate data are isotropic, which is often violated in real data due to differences in the covariance between variables. Thus, we extend NPDR to include a random forest distance, and other anisotropic distance metrics, for computing nearest neighbors. We also augment the isotope-ratio MS data with time-series features from the raw MS signal to improve biotic classification. We test NPDR on our novel experimental ocean world seawater analog MS data. We measure isotope fractionations of volatile CO 2 that could be measured in exospheres or plumes. Samples include baseline abiotic conditions using a range of possible seawater chemistry consistent with Europa and Enceladus, and biotic samples that include microbes in these seawaters. We use penalized NPDR with random forest proximity to identify interpretable microbial molecular signatures. We compare features with random forest importance, and we train a classifier that discriminates between biotic and abiotic samples with high accuracy. These ML-trained ocean-world analog MS data could be used to assist in identifying biosignatures during future missions.

geochemistry↗

Microfossils in the Antarctic cold desert: Possible implications for Mars

In the Ross Desert of Antarctica, the principal life form is the cryptoendolithic microbial community in the near-surface layers of porous sandstone rocks. Biological, geological, and climatic factors interact in a complex and precarious balance, making life possible in an otherwise hostile environment. Once this balance is tipped, fossilization sets in. In the reverse case, new colonization of the rock surface may be initiated. As a result, fossilization is contemporary with modern life and both may be simultaneously present in a mosaic pattern. Also, different stages of fossilization are present. The process of fossilization takes place in a nonaquatic environment. If primitive life ever appeared on Mars, it is possible that with increasing aridity, life withdrew into an endolithic niche similar to that in the Antarctic desert. Fossilization in a nonaquatic environment may have set in with the result that traces of past life could be preserved. If such was the case, the study of the fossilization process in Antarctica may hold useful information for the analysis of Martian samples for microfossils.

Friedmann, E. I.↗

Development of a preprototype hyperfiltration wash water recovery subsystem

The use of hyperfiltration as a mode of reclamation of waste water on board an extended mission spacecraft was investigated. Two basic approaches are considered with respect to hyperfiltration of wash water recovery. The initial approach involves the use of a hollow fiber permeator and a tubular module, operating at ambient temperature. In this system, relatively large doses of biocides are used to control microbial activity. Since biocides require a long contact time, and many have adverse dematological effects as well as many interact with membrane material, a second approach is considered which involves operating at pasturization temperature.

Source record↗

Spacelab 3 mission

Spacelab-3 (SL-3) was the first microgravity mission of extended duration involving crew interaction with animal experiments. This interaction involved sharing the Spacelab environmental system, changing animal food, and changing animal waste trays by the crew. Extensive microbial testing was conducted on the animal specimens and crew and on their ground and flight facilities during all phases of the mission to determine the potential for cross contamination. Macroparticulate sampling was attempted but was unsuccessful due to the unforseen particulate contamination occurring during the flight. Particulate debris of varying size (250 micron to several inches) and composition was recovered post flight from the Spacelab floor, end cones, overhead areas, avionics fan filter, cabin fan filters, tunnel adaptor, and from the crew module. These data are discussed along with solutions, which were implemented, for particulate and microbial containment for future flight facilities.

Dalton, Bonnie P.↗