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At least 73 records · Page 4
All sourcesNASA NTRSDOE OSTI
NASA NTRS2017

Stratosphere Conditions Inactivate Bacterial Endospores from a Mars Spacecraft Assembly Facility

Every spacecraft sent to Mars is allowed to land viable microbial bioburden, including hardy endospore-forming bacteria resistant to environmental extremes. Earth's stratosphere is severely cold, dry, irradiated, and oligotrophic; it can be used as a stand-in location for predicting how stowaway microbes might respond to the Martian surface. We launched E-MIST, a high-altitude NASA balloon payload on 10 October 2015 carrying known quantities of viable Bacillus pumilus SAFR-032 (4.07?×?10(7) spores per sample), a radiation-tolerant strain collected from a spacecraft assembly facility. The payload spent 8?h at ~31?km above sea level, exposing bacterial spores to the stratosphere. We found that within 120 and 240?min, spore viability was significantly reduced by 2 and 4 orders of magnitude, respectively. By 480?min, <0.001% of spores carried to the stratosphere remained viable. Our balloon flight results predict that most terrestrial bacteria would be inactivated within the first sol on Mars if contaminated spacecraft surfaces receive direct sunlight. Unfortunately, an instrument malfunction prevented the acquisition of UV light measurements during our balloon mission. To make up for the absence of radiometer data, we calculated a stratosphere UV model and conducted ground tests with a 271.1?nm UVC light source (0.5 W/m(2)), observing a similarly rapid inactivation rate when using a lower number of contaminants (640 spores per sample). The starting concentration of spores and microconfiguration on hardware surfaces appeared to influence survivability outcomes in both experiments. With the relatively few spores that survived the stratosphere, we performed a resequencing analysis and identified three single nucleotide polymorphisms compared to unexposed controls. It is therefore plausible that bacteria enduring radiation-rich environments (e.g., Earth's upper atmosphere, interplanetary space, or the surface of Mars) may be pushed in evolutionarily consequential directions.

Planetary protection↗
NASA NTRS2024

Lunar Soil Enrichment for Plant Production: WILD (Waste Improved Lunar Dirt)

To identify a process to generate a fertile soil for plant growth using lunar regolith and waste compost, several qualitative and quantitative investigations were performed. The primary focus of this project was to demonstrate the feasibility of microbe bioleaching to extract inorganic plant nutrients (P and K) from lunar regolith simulant. This was achieved by developing a screening test method to identify microbial candidates that would effectively bioleach P and K from JSC-1A. Quantitative bioleaching test using the downselected microbes (Pantoea Agglomerans (P. agglo) and a plant-relevant consortium) were then performed. The second focus of the project was to validate the approach of waste compost utilization to improve water retention and fertility of lunar regolith for food production. This was achieved by measuring the water retention property of the mixture of compost and regolith at different ratio and conducting microgreen growth experiments with compost/regolith mixtures. The team also discovered from this study that some native microbes associated with JSC-1A can form biofilm containing JSC-1A particles (a type of soil crust), which showed potential as an ISRU geomicrobiological dust mitigation method for Lunar or Martian applications.

Ray Pitts↗
NASA NTRS2010

Root Zone Respiration on Hydroponically Grown Wheat Plant Systems

Root respiration is a biological phenomenon that controls plant growth and physiological development during a plant's lifespan. This process is dependent on the availability of oxygen in the system where the plant is located. In hydroponic systems, where plants are submerged in a solution containing vital nutrients but no type of soil, the availability of oxygen arises from the dissolved oxygen concentration in the solution. This oxygen concentration is dependent on the , gas-liquid interface formed on the upper surface of the liquid, as given by Henry's Law, depending on pressure and temperature conditions. Respiration rates of the plants rise as biomass and root zone increase with age. The respiration rate of Apogee wheat plants (Triticum aestivum) was measured as a function of light intensity (catalytic for photosynthesis) and CO2 concentration to determine their effect on respiration rates. To determine their effects on respiration rate and plant growth microbial communities were introduced into the system, by Innoculum. Surfactants were introduced, simulating gray-water usage in space, as another factor to determine their effect on chemical oxygen demand of microbials and on respiration rates of the plants. It is expected to see small effects from changes in CO2 concentration or light levels, and to see root respiration decrease in an exponential manner with plant age and microbial activity.

Soler-Crespo, R. A.↗
NASA NTRS2016

Instrumentation for Examining Microbial Response to Changes In Environmental Pressures

The Automated Adaptive Directed Evolution Chamber (AADEC) is a device that allows operators to generate a micro-scale analog of real world systems that can be used to model the local-scale effects of climate change on microbial ecosystems. The AADEC uses an artificial environment to expose cultures of micro-organisms to environmental pressures, such as UV-C radiation, chemical toxins, and temperature. The AADEC autonomously exposes micro-organisms to selection pressures. This improves upon standard manual laboratory techniques: the process can take place over a longer period of time, involve more stressors, implement real-time adjustments based on the state of the population, and minimize the risk of contamination. We currently use UV-C radiation as the main selection pressure, UV-C is well studied both for its cell and DNA damaging effects as a type of selection pressure and for its related effectiveness as a mutagen; having these functions united makes it a good choice for a proof of concept. The AADEC roadmap includes expansion to different selection pressures, including heavy metal toxicity, temperature, and other forms of radiation.The AADEC uses closed-loop control to feedback the current state of the culture to the AADEC controller that modifies selection pressure intensity during experimentation, in this case culture density and growth rate. Culture density and growth rate are determined by measuring the optical density of the culture using 600 nm light. An array of 600 nm LEDs illuminate the culture and photodiodes are used to measure the shadow on the opposite side of the chamber.Previous experiments showed that we can produce a million fold increase to UV-C radiation over seven iterations. The most recent implements a microfluidic system that can expose cultures to multiple different selection pressures, perform non-survival based selection, and autonomously perform hundreds of exposure cycles. A scalable pump system gives the ability to pump in various different growth media to individual cultures and introduce chemical toxins during experimentation; AADEC can perform freeze and thaw cycles. We improved our baseline characterization by building a custom UV-C exposure hood, a shutter operates on a preset timer allowing the user to set exposure intensity consistently for multiple iterations.

Environment↗
NASA NTRS2002

Composition and physiological profiling of sprout-associated microbial communities

The native microfloras of various types of sprouts (alfalfa, clover, sunflower, mung bean, and broccoli sprouts) were examined to assess the relative effects of sprout type and inoculum factors (i.e., sprout-growing facility, seed lot, and inoculation with sprout-derived inocula) on the microbial community structure of sprouts. Sprouts were sonicated for 7 min or hand shaken with glass beads for 2 min to recover native microfloras from the surface, and the resulting suspensions were diluted and plated. The culturable fraction was characterized by the density (log CFU/g), richness (e.g., number of types of bacteria), and diversity (e.g., microbial richness and evenness) of colonies on tryptic soy agar plates incubated for 48 h at 30 degrees C. The relative similarity between sprout-associated microbial communities was assessed with the use of community-level physiological profiles (CLPPs) based on patterns of utilization of 95 separate carbon sources. Aerobic plate counts of 7.96 +/- 0.91 log CFU/g of sprout tissue (fresh weight) were observed, with no statistically significant differences in microbial cell density, richness, or diversity due to sprout type, sprout-growing facility, or seed lot. CLPP analyses revealed that the microbial communities associated with alfalfa and clover sprouts are more similar than those associated with the other sprout types tested. Variability among sprout types was more extensive than any differences between microbial communities associated with alfalfa and clover sprouts from different sprout-growing facilities and seed lots. These results indicate that the subsequent testing of biocontrol agents should focus on similar organisms for alfalfa and clover, but alternative types may be most suitable for the other sprout types tested. The inoculation of alfalfa sprouts with communities derived from various sprout types had a significant, source-independent effect on microbial community structure, indicating that the process of inoculation alters the dynamics of community development regardless of the types of organisms involved.

NASA Center KSC↗
NASA NTRSDate not supplied

SPACEFLIGHT-INDUCED CHANGES IN MICROBIAL VIRULENCE AND THE IMPACT TO THE HOST IMMUNE RESPONSE

INTRODUCTION Over the past 50 years, microorganisms have displayed 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, and virulence. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system; consisting of altered leukocyte distribution, reductions in T and NK cell function, altered cytokine profiles, and reactivation of latent herpesviruses. Further, astronauts have some degree of clinical incidence, primarily infectious disease episodes and atopic dermatitis. The impact of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. SPECIFIC AIMS In the Host-Microorganism, Microbial Response aspect of this study, the following Specific Aims are being investigated, using the microbial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli. Aim 1: Characterize the effect of spaceflight analogue culture on microbial pathogenesis-related stress responses and in vitro host-pathogen interactions. Analysis will include microbial stress responses as well as colonization and viability following pathogen challenge of three-dimensional (3-D) tissue co-culture models containing immune cells. We specifically selected obligate and opportunistic pathogens that are medically important and have been or are likely to be found aboard spacecraft. Aim 2: Characterize the effect of spaceflight analogue culture on the virulence potential of pathogenic microorganisms. Changes in virulence will be assessed using a mouse model of infection. SIGNIFICANCE The goal of this study is to use spaceflight analogue conditions to gain insight into the breadth of medically-significant International Space Station microorganisms that have altered virulence and the impact of those changes on the immune response of the host, including astronaut immune cells. This information will provide critical understanding into the impact of microgravity on potential alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.

C. M. Ott↗
NASA NTRSDate not supplied

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

INTRODUCTION Over the past 50 years, many microorganisms have displayed unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in stress resistance, biofilm production, antibiotic sensitivity, final cell concentration, gene expression, enhanced host-pathogen interaction, and virulence. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system; consisting of altered leukocyte distribution, reductions in T and NK cell function, altered cytokine profiles, and reactivation of latent herpesviruses. Further, astronauts have some degree of clinical incidence, primarily infectious disease episodes and atopic dermatitis. The impact of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. SPECIFIC AIMS In this study, the following Specific Aims are being investigated, using the microbial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli (EHEC). Aim 1: Characterize the effect of spaceflight analogue culture on microbial pathogenesis-related stress responses and in vitro host-pathogen interactions. Analyses include microbial stress responses as well as colonization and viability following pathogen challenge of three-dimensional (3-D) tissue co-culture models containing immune cells. We specifically selected obligate and opportunistic pathogens that are medically important and have been or are likely to be found aboard spacecraft. Aim 2: Characterize the effect of spaceflight analogue culture on the virulence potential of pathogenic microorganisms. Virulence will be assessed using a mouse model of infection. SIGNIFICANCE The goal of this study is to use spaceflight analogue conditions to gain insight into the breadth of medically-significant International Space Station microorganisms that have altered virulence and the impact of those changes on the immune response of the host. This information will provide critical understanding into the impact of microgravity on potential alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.

C M Ott↗
NASA NTRS2017

A PCR Based Microbial Monitoring Alternative Method of Detection and Identification of Microbes Aboard ISS

Previous research has shown that microorganisms and potential human pathogens have been detected on the International Space Station (ISS) with additional introduction of new microflora occurring with every exchange of crew or addition of equipment and supplies. These microbes are readily transferred between crew and subsystems (i.e. ECLSS, environmental control and life support systems). As this can be detrimental to astronaut health and optimal performance of ISS systems, monitoring of systems such as ECLSS to include identification of microbial contaminants could prevent adverse effects on human health and life support systems. Current monitoring on ISS is laborious and utilizes culture based methods followed by sample return to Earth for complete analysis. Future, long-distance spaceflight missions will require real-time monitoring capabilities that enable efficient and rapid assessments of the microbial environment allowing for expedited decisions and more targeted response to cope with anomalies. Polymerase chain reaction (PCR), a molecular microbial monitoring method was chosen and numerous PCR instruments investigated for their potential to perform in microgravity conditions. Using ISS as a test bed for PCR verification in microgravity will enable NASA to assess whether molecular based microbiological sensors may be components of reliable, closed-loop life support and habitation systems in spacecraft, enhancing infrastructure capabilities through increased efficiency, reliability, and time savings by enabling sample analysis on orbit. NASA selected the Water Monitoring Suite as one of the rapid spaceflight hardware demonstration activities utilizing a streamlined process to minimize the time required to fly experimental flight hardware. The RAZOR EX (BioFire Defense, Salt Lake City, UT) system was part of the water monitoring suite and is a commercial off-the-shelf (COTS) real-time PCR instrument designed for field work. The RAZOR EX was originally designed for Department of Defense (DoD) under a small business innovative research (SBIR) grant and is ruggedized, compact and provides a rapid, sample to answer in less than an hour. PCR assays using a fluorescent probe were optimized and spiked with known concentrations of DNA (Pseudomonas aeruginosa) ranging from 0.002 to 20 ng. PCR reagents were lyophilized and configured in customized pouches and tested for flight readiness. Three types of water were used to rehydrate the reagents and demonstrate the fidelity of the PCR reaction in microgravity. Molecular grade deionized water served as a control while filtered and unfiltered ISS potable water served to test for chemical or biological inhibitors. All three types were compared to parallel ground test results. Nine tests were run on ISS (3 of each water type) and the critical threshold cycle (Ct) was compared to parallel ground tests completed at Kennedy Space Center, FL and Johnson Space Center, TX. All concentrations of Pseudomonas aeruginosa DNA were detected. A comparison of the Ct produced in real time PCR indicated similarity between flight and ground samples. There appeared to be no significant difference between flight or ground PCR reactions or between any of the three water types. This testing demonstrated the ability to perform molecular testing during spaceflight operations with similar sensitivity. It will allow for future ground development of molecular protocols and minimize the need for spaceflight testing. Future testing will include development of additional targets including environmental and health related organisms.

Christina Khodadad↗
NASA NTRS2002

Long Term Manipulations of Intact Microbial Mat Communities in a Greenhouse Collaboratory: Simulating Earth's Present and Past Field Environments

Photosynthetic microbial mat communities were obtained from marine hypersaline saltern ponds, maintained in a greenhouse facility, and examined for the effects of salinity variations. Because these microbial mats are considered to be useful analogs of equivalent ancient marine communities, they offer insights about evolutionary events during the greater than 3 billion year time interval wherein mats co-evolved with Earth's geosphere and atmosphere. Although photosynthetic mats can be highly dynamic and exhibit extremely high activity, the mats in the present study have been maintained for more than one year with relatively minor changes. The major groups of microorganisms, as assayed using microscopic, genetic, and biomarker methodologies, are essentially the same as those in the original field samples. Field and greenhouse mats were similar with respect to rates of exchange of oxygen and dissolved inorganic carbon across the mat-water interface, both during the day and at night. Field and greenhouse mats exhibited similar rates of efflux of methane and hydrogen. Manipulations of salinity in the water overlying the mats produced changes in the community that strongly resemble those observed in the field. A collaboratory testbed and an array of automated features are being developed to support remote scientific experimentation with the assistance of intelligent software agents. This facility will permit teams of investigators to explore ancient environmental conditions that are rare or absent today but might have influenced the early evolution of these photosynthetic ecosystems.

Bebout, Brad↗
NASA NTRSDate not supplied

Microbial Methods for Testing a Novel Spacecraft Bioburden Reduction Method

In order to circumvent the contamination of microorganisms in both extraterrestrial and terrestrial environments, spacecraft hardware must often be sterilized. Current sterilization practices are both time-consuming and expensive. Techniques such as dry heat sterilization and scattered/localized electric field enhancements are incompatible with a variety of hardware materials as well as complex surface shapes. Our group is testing a novel femtosecond pulsed laser for the rapid sterilization of spacecraft hardware using high photon fluxes, which is both an efficient and effective technique to ensure a reduced microbial burden aboard spacecraft. This method is likely to be effective because microorganisms have not encountered high photon fluxes in their evolutionary history and thus contain no protective mechanisms against them. To assess the effectiveness of pulsed laser sterilization, we used a combination of scanning electron microscopy and elemental analysis for the examination of damaged spores on metal coupons, as well as microbiology methods for spore assays. The surfaces of metal coupons inoculated with Bacillus subtilis were illuminated with femtosecond pulses at various wavelengths below the damage threshold for the aluminum. After laser treatment, metal coupon samples were sonicated to dislodge spores, serially diluted, and finally plated and incubated. Results show that sterilization efficiency is favorable in some conditions. The NASA standards for spore assays are a preliminary demonstration in establishing the trend in ability to kill spores. The results show potential for the dual pulse illumination method as a sterilization technique for planetary protection applications.

Maha S Ulhaq↗
NASA NTRS1975

The Apollo 16 microbial response to space environment experiment

The effect was evaluated of a particular space flight on the survival rate of nine different species. Although a reasonable variety of organisms (viruses, yeasts, filamentous fungi, bacteria, and an invertebrate) were tested under several different conditions, no statistically valid differences could be detected in the survival of flight samples when compared to corresponding ground-based controls. In general, these evaluations were based on multiple observations of from ten to thirty replicates of up to one million cells each. While the results conflict with those of certain other space flight investigations, it is observed that the conditions of a particular space flight cannot be exactly duplicated, and therefore results from different flights are not directly comparable.

Gerald R. Taylor↗
NASA NTRSDate not supplied

NASA SmallSat LEARN Forum Briefing: BioSentinel

We will focus on the technologies developed for BioSentinel, primarily the science payloads. One of them, the BioSensor, is a microfluidics-based instrument that houses a series of fluidic cards and manifolds. The fluidic cards can be loaded with microbial organisms to study the effect of the space environment on biology. For BioSentinel, we used the budding yeast as our model organism to study the response to the radiation environment in both low Earth orbit (ISS) and in deep space. The second instrument is a radiation spectrometer. This sensor allows the quantification of ionizing radiation as well as the characterization of the different particles based on the linear energy transfer or LET. Given that BioSentinel launched this past November (Artemis I), part of the presentation will include lessons learned and how we will apply them to future missions, including LEIA. LEIA was recently selected for a mission to the lunar surface on a CLPS lander vehicle, and is primarily based at NASA Ames.

BioSentinel↗
NASA NTRS2024

Space Environmental Effects on Cleanroom Microbes: A Pathfinder Study

Microbes have an extraordinary ability to adapt to their environment, but little is known about the effects of space irradiation on microbial survival and adaptation. Microbial responses are relevant to both Planetary Protection (PP), where microbes could contaminate planetary bodies and inhibit scientific investigation, and in crew wellbeing where microbes may develop unknown, dangerous resistance mechanisms that impact human health. This project aimed to determine survivability of a collection of isolates collected from NASA Marshall Space Flight Center (MSFC) cleanrooms. In this study, we developed a standard process for exposing dried microbes to various levels of proton irradiation, followed by evaluation of survival. Through multiple exposures at various intensities, we were able to narrow down a shortlist of cleanroom microbes deemed of highest threat to both PP and crew health. These findings could help to more clearly and accurately define risks associated with radiation impacts on microbes.

Planetary Protection↗
NASA NTRSDate not supplied

Harnessing Synthetic Communities and Microbial Recycling of Space Waste Streams for Biomanufacturing Applications

The long-term habitation of extraterrestrial environments such as the Moon or Mars presents significant challenges including supplying materials to sustain life. Off-world recycling of waste materials into biomanufacturer products may ameliorate this. Current literature highlights the need for efficient waste recycling systems to support the bioproduction of essential materials including foods, pharmaceuticals, and biomaterials. The study herein concerns itself with the investigation of three key aspects: 1) formulating an optimal wastewater media on which to grow recombinant microbes for bioproduction in space, 2) examining the potential for constructing stable and metabolically synergistic synthetic microbial communities for largescale and multi-tiered biomanufacturing, and 3) testing the efficacy of one such bioengineered enzyme, cutinase, on the degradation of PET plastics characteristic of those found in ISS waste as a model for recombinant recycling-based biomanufacturing of mission-critical substrates. Formulation of an optimal wastewater media involved growing several microorganisms on mixtures of synthetic planetary wastes representative of those found in space waste systems, combined with simple carbon sources derived from a physio-chemical CO conversion system to determine their growth potential. Potential synthetic microbial communities were conceptually designed, and their stability and metabolic synergism was evaluated within the context of co-cultures. Cutinase activity assays were utilized to determine the efficacy of bioengineered cutinase on PET plastic degradation. Findings will contribute to optimization of wastewater-based media formulations, data on stable synthetic microbial communities for bioproduction, and effective methods for measuring cutinase-based PET plastic degradation. These outcomes support the development of sustainable waste recycling systems for space habitation and aim to fill gaps in the current literature and proposing innovative solutions for waste recycling in space environments. By leveraging synthetic biology this study seeks to enhance the feasibility of long-term extraterrestrial habitation through sustainable resource management.

Bioprocessing↗
NASA NTRS1973

Weightlessness

Weightlessness effects on physiological systems are summarized, including cardiovascular responses, hematological effects, work capacity (exercise tolerance), respiratory responses, skeletal responses, neuromuscular changes, vestibular responses, metabolism, endocrine and electrolyte responses, and weight loss. The nature of the adaptive response to space flight and microbial changes are also discussed. The effects of zero gravity upon performance are ascertained, and the physiological effects of weightlessness are counteracted.

Charles Berry↗
NASA NTRS1991

Microbial growth and physiology in space - A review

An overview of microbial behavior in closed environments is given with attention to data related to simulated microgravity and actual space flight. Microbes are described in terms of antibiotic sensitivity, subcellular structure, and physiology, and the combined effects are considered of weightlessness and cosmic radiation on human immunity to such microorganisms. Space flight results report such effects as increased phage induction, accelerated microbial growth rates, and the increased risk of disease communication and microbial exchange aboard confining spacecraft. Ultrastructural changes are also noted in the nuclei, cell membranes, and cytoplasmic streaming, and it appears that antibiotic sensitivity is reduced under both actual and simulated conditions of spaceflight.

Cioletti, Louis A.↗