Results for “microbial growth system”

NASA NTRS2016

PowerCells Payload on EuCROPIS - Measuring Synthetic Biology in Space

NASA’s PowerCell payload on the DLR (Deutsches Zentrum fur Luft- und Raumfahrt, i.e. German Aerospace Center) Eu:CROPIS satellite will compare the effect of multiple simulated gravity regimes on basic processes required for synthetic biology in space including growth, protein production, and genetic transformation of the bacterium Bacillus subtilis. In addition, it will pioneer the use of a cyanobacterially-produced feedstock for microbial growth in space, a concept we call “PowerCell.” The PowerCell experiment system will be integrated using the Spaceflight Secondary Payload System with the German Space Agency's (DLR's) Euglena and Combined Regenerative Organic-food Production In Space (Eu:CROPIS) satellite, to be launched during the summer of 2017. In order to simulate the gravitational gradient of different celestial bodies, the Eu:CROPIS satellite will establish artificial microgravity, lunar, and Martian gravity levels prior to conducting each set of biological experiments, with experimental results compared to ground controls. Experiments will be carried out in microfluidics cards with experimental progress measured through absorbance as detected by the LED-based optical system. Here we describe the ground studies that led to these experiments, along with a description of the flight hardware and its performance. The results of this mission will provide foundational data for the use and production of genetically engineered organisms for extraterrestrial missions.

PowerCell experiment system↗

NASA NTRSDate not supplied

Remodeling BioSentinel’s Microbial Metabolism Measurement System

BioSentinel, the first biological CubeSat beyond low Earth orbit, will investigate how Saccharomyces cerevisiae responds to ionizing radiation using optical measurements of a colorimetric metabolic indicator dye (alamarBlue). To map optical data to more detailed metabolic measurements, a microbial culture system for ground testing was designed consisting of a 400 mL beaker and a rubber stopper with pass-through holes for its five sensor probes (electrical conductivity, pH, oxidation-reduction potential, dissolved oxygen, optical density). The initial design had several drawbacks. It was only partially reusable due to its one-time sealants: Teflon tape and parafilm. A steady increase in oxygen levels within the sensor’s testingdata indicated the design was not sufficiently airtight. Degradation of the rubber stopper’s elasticity due to sterilization and reinsertion of the probes led to poor robustness and eventual replacement. The vessel’s large headspace didn’t reproduce flight conditions and a large culture volume required more reagents. The system has been redesigned to contain a rigid, biocompatible polycarbonate disc lid that allows for threaded inserts. The culture vessel’s height will be reduced from 110 mm to 65 mm to decrease overhead space and material costs. A more reliable seal will be achieved with a gasket and clamps. The optical probe records absorbance at four wavelengths to characterize cell density and alamarBlue dye ratios. A pneumatically operated plunger draws media through the hollow interior of the probe. Currently, deformation of the silicone on the plunger causes asymmetrical movement and a drift from its initial state. Designs are being developed to limit the plunger’s travel for a more accurate measurement cycle. The remodeled system is under construction and will be tested for leaks and accuracy. The measurements collected will be used to provide additional insight into the optical data returned from the BioSentinel mission.

Astrobiology↗

NASA NTRS2010

International Space Station (ISS) Internal Active Thermal Control System (IATCS) New Biocide Selection, Qualification and Implementation

The Internal Active Thermal Control System (IATCS) aboard the International Space Station (ISS) is primarily responsible for the removal of heat loads from payload and system racks. The IATCS is a water based system which works in conjunction with the EATCS (External ATCS), an ammonia based system, which are interfaced through a heat exchanger to facilitate heat transfer. On-orbit issues associated with the aqueous coolant chemistry began to occur with unexpected increases in CO2 levels in the cabin. This caused an increase in total inorganic carbon (TIC), a reduction in coolant pH, increased corrosion, and precipitation of nickel phosphate. These chemical changes were also accompanied by the growth of heterotrophic bacteria that increased risk to the system and could potentially impact crew health and safety. Studies were conducted to select a biocide to control microbial growth in the system based on requirements for disinfection at low chemical concentration (effectiveness), solubility and stability, material compatibility, low toxicity to humans, compatibility with vehicle environmental control and life support systems (ECLSS), ease of application, rapid on-orbit measurement, and removal capability. Based on these requirements, ortho-phthalaldehyde (OPA), an aromatic dialdehyde compound, was selected for qualification testing. This paper presents the OPA qualification test results, development of hardware and methodology to safely apply OPA to the system, development of a means to remove OPA, development of a rapid colorimetric test for measurement of OPA, and the OPA on-orbit performance for controlling the growth of microorganisms in the ISS IATCS since November 3, 2007.

Wilson, Mark E.↗

NASA NTRS2011

International Space Station (ISS) Internal Active Thermal Control System (IATCS) New Biocide Selection, Qualification and Implementation

The Internal Active Thermal Control System (IATCS) aboard the International Space Station (ISS) is primarily responsible for the removal of heat loads from payload and system racks. The IATCS is a water based system which works in conjunction with the EATCS (External ATCS), an ammonia based system, which are interfaced through a heat exchanger to facilitate heat transfer. On-orbit issues associated with the aqueous coolant chemistry began to occur with unexpected increases in CO2 levels in the cabin. This caused an increase in total inorganic carbon (TIC), a reduction in coolant pH, increased corrosion, and precipitation of nickel phosphate. These chemical changes were also accompanied by the growth of heterotrophic bacteria that increased risk to the system and could potentially impact crew health and safety. Studies were conducted to select a biocide to control microbial growth in the system based on requirements for disinfection at low chemical concentration (effectiveness), solubility and stability, material compatibility, low toxicity to humans, compatibility with vehicle environmental control and life support systems (ECLSS), ease of application, rapid on-orbit measurement, and removal capability. Based on these requirements, ortho-phthalaldehyde (OPA), an aromatic dialdehyde compound, was selected for qualification testing. This paper presents the OPA qualification test results, development of hardware and methodology to safely apply OPA to the system, development of a means to remove OPA, development of a rapid colorimetric test for measurement of OPA, and the OPA on-orbit performance for controlling the growth of microorganisms in the ISS IATCS since November 3, 2007.

Wilson, Mark E.↗

DOE OSTI2020

An Arduino based automatic pressure evaluation system to quantify growth of non‐model anaerobes in culture

Abstract Measuring the growth rate of non‐model anaerobic microbes typically requires the use of time‐consuming and often destructive manual measurements. Here, an Arduino based automatic pressure evaluation system (A‐APES) was developed to automatically measure the rate of fermentation gas production as a proxy for microbial growth in anaerobic systems. The A‐APES system measures accumulated gas pressure in sealed cultures accurately at high‐resolution, while venting the system at programmed intervals to prevent over pressurization. The utility of A‐APES is demonstrated in this study by quantifying the growth rate and phases of a biomass‐degrading anaerobic gut fungus, which cannot be otherwise measured via conventional techniques due to its association with particulate substrates. Given the utility of the A‐APES approach, we provide a complete construction guide to fabricate the device, which is three times less expensive compared to existing commercial alternatives.

Wilken, St. Elmo↗

NASA NTRSDate not supplied

To Biocide or not to Biocide? Exploring the "No Biocide" Option in Spacecraft Potable Water Systems

Residual biocide has been used to protect against microbial growth in spacecraft potable water systems since the Gemini program. Iodine, the biocide currently used on the International Space Station (ISS) has a long history of use but presents a few challenges for exploration: it must be removed using a consumable absorbent before crew consumption and is known to lose its biocidal concentration due to interactions with wetted components, especially during periods of dormancy. Biocidal silver is being considered for exploration missions as it is safe for crew consumption, but it too has challenges with proper material design to maintain sufficient disinfection residuals. An idea proposed by some in the Environmental Control and Life Support System (ECLSS) community has been that the biocide does not provide a true barrier against microbial growth, and therefore, perhaps the challenges of maintaining a residual biocide in the system could be best overcome by removing the biocide altogether. In this argument, they point to the regenerative water treatment system on the ISS which produces high purity water that theoretically meets the microbiological requirements before the iodine is added. Exploration water processing systems are expected to have similar water processing capabilities. This paper will discuss the role that residual biocide may play in controlling microbial growth and presents perspectives on the potential risks associated with removing it from exploration water systems.

biocide↗

NASA NTRSDate not supplied

To Biocide or not to Biocide? Exploring the "No Biocide" Option in Spacecraft Potable Water Systems

Residual biocide has been used to protect against microbial growth in spacecraft potable water systems since the Gemini program. Iodine, the biocide currently used on the International Space Station (ISS) has a long history of use but presents a few challenges for exploration: it must be removed using a consumable absorbent before crew consumption and is known to lose its biocidal concentration due to interactions with wetted components, especially during periods of dormancy. Biocidal silver is being considered for exploration missions as it is safe for crew consumption, but it too has challenges with proper material design to maintain sufficient disinfection residuals. An idea proposed by some in the Environmental Control and Life Support System (ECLSS) community has been that the biocide does not provide a true barrier against microbial growth, and therefore, perhaps the challenges of maintaining a residual biocide in the system could be best overcome by removing the biocide altogether. In this argument, they point to the regenerative water treatment system on the ISS which produces high purity water that theoretically meets the microbiological requirements before the iodine is added. Exploration water processing systems are expected to have similar water processing capabilities. This paper will discuss the role that residual biocide may play in controlling microbial growth and presents perspectives on the potential risks associated with removing it from exploration water systems.

biocide↗

NASA NTRSDate not supplied

To Biocide or Not to Biocide? Exploring the "No Biocide" Option in Spacecraft Potable Water Systems

Residual biocide has been used to protect against microbial growth in spacecraft potable water systems since the Gemini program. Iodine, the biocide currently used on the International Space Station (ISS) has a long history of use but presents a few challenges for exploration: it must be removed using a consumable absorbent before crew consumption and is known to lose its biocidal concentration due to interactions with wetted components, especially during periods of dormancy. Biocidal silver is being considered for exploration missions as it is safe for crew consumption, but it too has challenges with proper material design to maintain sufficient disinfection residuals. An idea proposed by some in the Environmental Control and Life Support System (ECLSS) community has been that the biocide does not provide a true barrier against microbial growth, and therefore, perhaps the challenges of maintaining a residual biocide in the system could be best overcome by removing the biocide altogether. In this argument, they point to the regenerative water treatment system on the ISS which produces high purity water that theoretically meets the microbiological requirements before the iodine is added. Exploration water processing systems are expected to have similar water processing capabilities. This paper will discuss the role that residual biocide may play in controlling microbial growth and presents perspectives on the potential risks associated with removing it from exploration water systems.

biocide↗

NASA NTRSDate not supplied

To Biocide or Not to Biocide? Exploring the "No Biocide" Option in Spacecraft Potable Water Systems

Residual biocide has been used to protect against microbial growth in spacecraft potable water systems since the Gemini program. Iodine, the biocide currently used on the International Space Station (ISS) has a long history of use but presents a few challenges for exploration: it must be removed using a consumable absorbent before crew consumption and is known to lose its biocidal concentration due to interactions with wetted components, especially during periods of dormancy. Biocidal silver is being considered for exploration missions as it is safe for crew consumption, but it too has challenges with proper material design to maintain sufficient disinfection residuals. An idea proposed by some in the Environmental Control and Life Support System (ECLSS) community has been that the biocide does not provide a true barrier against microbial growth, and therefore, perhaps the challenges of maintaining a residual biocide in the system could be best overcome by removing the biocide altogether. In this argument, they point to the regenerative water treatment system on the ISS which produces high purity water that theoretically meets the microbiological requirements before the iodine is added. Exploration water processing systems are expected to have similar water processing capabilities. This paper will discuss the role that residual biocide may play in controlling microbial growth and presents perspectives on the potential risks associated with removing it from exploration water systems.

biocide↗

NASA NTRSDate not supplied

Spaceflight Autonomous Multigenerational Microbial Sequencer in Support of Plant-Growth Systems

The CubeSat platform has proven successful in obtaining meaningful life science information when biological payloads are incorporated. Examples include: 1) the first-ever CubeSat with a biological payload, GeneSat-1, which demonstrated decreased growth rates for flight samples of Escherichia coli in low Earth orbit (Parra et al. 2008); 2) PharmaSat, demonstrated that Saccharomyces cerevisiae in the microgravity environment exhibits a significant level of metabolic activity even at high doses of applied antifungal (Ricco et al. 2011); 3) O/OREOS, which used Bacillus subtilis(bacteria) to demonstrate for the first time that microorganisms can be loaded in a dried, dormant form and then rehydrated and grown in orbit months after launch (Nicholson et al. 2011; Ehrenfreund et al. 2014; 4) the SporeSat payload, which investigated Ceratopteris richardii(fern spores) using lab-on-a-chip devices (BioCDs) and minicentrifuges to produce artificial gravitational forces in ground studies (Park et al. 2017), with demonstration of the BioCD and minicentrifuge in space; 5) EcAMSat, the first CubeSat to be directly deployed from the ISS for an experiment assessing antibiotic resistance of E. coli in the microgravity environment (Padgen et al. 2020); 6) BioSentinel, exposed a culture of yeast to galactic cosmic radiation (GCR) and solar particle events while in heliocentric orbit to measure the rate of double-strand-break repair using DNA-repair-deficient mutants. This effort measures the metabolic parameters of yeast in a deep-space environment compared to Earth ambient conditions using a 3-color LED detection system (Ricco et al. 2020; Padgen et al. 2021). We aim to expand this list to include a Spaceflight Autonomous Multigenerational Microbial Sequencer (SAMMS). SAMMS will allow for the genome level understanding of changes in growth and metabolic activity for any organism. While microbes are suitable for early studies in our proposed platform because of their small size, small and relatively less-complicated genomes, fast generation times, and relevance to life support systems; multicellular organisms can similarly be evaluated for their genetic response to the spaceflight environment. The Spaceflight Autonomous Multigenerational Microbial Sequencer (SAMMS) will enable autonomous sequencing of biological samples in plant production units, cislunar orbit and on the lunar surface to examine spaceflight effects (ie. radiation, altered gravity, reduced pressures) on plant and microbial genomes.On this team a Kennedy Space Center (KSC) space crop production and water systems microbiologist/molecular biologist works with a Johnson Space Center (JSC) International Space Station (ISS) microbial sequencing expert and an Ames Research Center (ARC) CubeSat Engineering team to convert an automated Oxford Nanopore librarypreparation and sequencing method to a fluidic CubeSat payload system. The Oxford Nanopore MinION sequencing platform has proven successful in the spaceflight environment onboard the ISS (Stahl-Rommel et al. 2021). Further long-duration spaceflight and exposure to high levels of radiation will cause genotypic effects in biological organisms that may affect their function. Monitoring the adaption of a population to the spaceflight environment and any subsequent beneficial mutations will allow for the harnessing of organisms best suited for use in life support systems. This will ensure that the selected life support-essential microorganisms maintain their intended specified function over generations of culturing in the relevant spaceflight environment without becoming hazardous to crew or spacecraft systems.

Aubrie E Orourke↗

NASA NTRSDate not supplied

Testing and validation of the microbial environment of the NASA rodent spaceflight habitat water delivery system.

Sterilized, deionized water within a closed, self-sufficient system has been used in NASA spaceflight rodent studies for several decades. Within the specialized spaceflight rodent habitat, water is delivered through a compression spring-loaded bag system to maintain positive pressure. Refill of the drinking water occurs every 30 days by direct transfer of potable water from aboard the International Space Station (ISS). This enables long term use without a weekly water change out, which meets spaceflight requirements, but contrasts with the general guidelines for the sanitation of water delivery systems. Even though the water is iodinated to minimize microbial growth, rodents are fed a special diet of high moisture nutrient-rich food bars based on the AIN-93 diet that may contribute to microbial growth in this water system. We designed a ground study to assess the quality of drinking water that is given to rodents throughout a mission. We conducted the ground test using 20 female C57BL/6J mice housed in this specialized habitat to mimic the timeline of a 90-day mission as well as the environmental conditions (temperature, humidity, and pCO2) within the ISS. We used a novel sampling method to test water at 2-week intervals for the 90-days, and also after each 30-day refill of the water delivery system. Mice in standard vivarium cages with water bottles were also included for comparison to the habitat. The results showed that overall microbial load remained close to zero for the duration, while total organic compound concentrations increased from 1370g/L to 10650g/L over the course of 90 days but remained below the level of concern. Inorganic ions and pH were also found to be at acceptable levels. Overall, we conclude that this system is effective in delivering clean, potable water to rodents for the 90-day duration of current missions to the ISS.

water↗

NASA NTRS1996

Engineering strategies for the design of plant nutrient delivery systems for use in space: approaches to countering microbiological contamination

Microbiological contamination of crops within space-based plant growth research chambers has been postulated as a potentially significant problem. Microbial infestations; fouling of Nutrient Delivery System (NDS) fluid loops; and the formation of biofilms have been suggested as the most obvious and important manifestations of the problem. Strict sanitation and quarantine procedures will reduce, but not eliminate, microbial species introduced into plant growth systems in space habitats. Microorganisms transported into space most likely will occur as surface contaminants on spacecraft components, equipment, the crew, and plant-propagative materials. Illustrations of the potential magnitude of the microbiological contamination issue will be drawn from the literature and from documentation of laboratory and commercial field experience. Engineering strategies for limiting contamination and for the development of countermeasures will be described. Microbiological control technologies and NDS hardware will be discussed. Configurations appropriate for microgravity research facilities, as well as anticipated bio-regenerative life support system implementations, will be explored. An efficiently designed NDS, capable of adequately meeting the environmental needs of crop plants in space, is considered to be critical in both the research and operational domains. Recommended experiments, tests, and technology developments, structured to allow the development of prudent engineering solutions also will be presented.

NASA Discipline Number 61-10↗

DOE OSTI2020

Visualizing Microbial Community Dynamics via a Controllable Soil Environment

Visualizing microbial community dynamics within soil environments remains challenging. Here, we developed a custom-built SoilBox system that facilitates probing various changes occurring in microbial community development and molecular processes when exposed to environmental perturbations (e.g., temperature changes, drought, nutrient availability). Using confocal microscopy along with mass spectrometry imaging, we demonstrate the power of this system by correlating microbial growth and spatial metabolic information from soil communities under different moisture regimes.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS2019

Spacecraft Lighting to Mitigate Microbial Growth

ISS crew health research indicate risk for immune system degradation for long duration missions, necessitating the need to maximize countermeasures on microbial growth (1). This project will investigate applications using violet (400nm) LEDs to mitigate growth of bacteria on architectural surfaces. The application is primarily for architectures and surfaces where a higher amount of bacterial growth is expected. The project will build two different types of violet light sources that represent lamp types that could be installed in spacecraft applications. Under a controlled setting, using violet LEDs, the evaluation will test the effectiveness of a glowing work surface using LED Panel verses an LED array overhead lamp. The effectiveness of the lighting systems to attenuate bacterial growth will be compared to a control. The results of this study can be used to inform spacecraft system architects on novel ways to improve habitat health, and reduce reliance on manual cleaning methods.

Clark, Toni A.↗

NASA NTRS1992

Coupling plant growth and waste recycling systems in a controlled life support system (CELSS)

The development of bioregenerative systems as part of the Controlled Ecological Life Support System (CELSS) program depends, in large part, on the ability to recycle inorganic nutrients, contained in waste material, into plant growth systems. One significant waste (resource) stream is inedible plant material. This research compared wheat growth in hydroponic solutions based on inorganic salts (modified Hoagland's) with solutions based on the soluble fraction of inedible wheat biomass (leachate). Recycled nutrients in leachate solutions provided the majority of mineral nutrients for plant growth, although additions of inorganic nutrients to leachate solutions were necessary. Results indicate that plant growth and waste recyling systems can be effectively coupled within CELSS based on equivalent wheat yield in leachate and Hoagland solutions, and the rapid mineralization of waste organic material in the hydroponic systems. Selective enrichment for microbial communities able to mineralize organic material within the leachate was necessary to prevent accumulation of dissolved organic matter in leachate-based solutions. Extensive analysis of microbial abundance, growth, and activity in the hydroponic systems indicated that addition of soluble organic material from plants does not cause excessive microbial growth or 'biofouling', and helped define the microbially-mediated flux of carbon in hydroponic solutions.

Garland, Jay L.↗

NASA NTRSDate not supplied

Spaceflight Autonomous Multigenerational Microbial Sequencer (SAMMS) in Support of Plant-Growth Systems

Explore the source record for details and available documents.

Cubesat↗

NASA NTRS2023

Spaceflight Autonomous Multigenerational Microbial Sequencer (SAMMS) in Support of Plant-Growth Systems

As the National Aeronautics and Space Association (NASA) begins to pursue long-duration space flights, they will need to be able to provide astronauts with a nutritious and reliable food source. To meet the administration’s goal of traveling to the Moon and Mars, astronauts will need to begin to grow their own food in space. To protect their food source, extensive monitoring will occur to test for the effects of a space flight environment (e.g., radiation) as well as for early pathogen and disease detection. Genomic sequencing allows for both concerns to be tested on a regular basis. However, NASA’s current sequencer is unable to process plant tissues. Therefore, a novel method for plant DNA extraction using microneedle (MN) patches that will be able to feed into NASA’s existing system, but also require minimal human input is proposed. To support this, the design was broken down into four components (1) MN patch fabrication (2) MN patch extraction, (3) automated sampling motion control, and (4) a processing module. The MN patch is fabricated using a custom mold with conically shaped needles. The mold is filled with Polyvinyl alcohol (PVA) solution and placed in a vacuum desiccator. The mold is left in the vacuum overnight until the patch is dry and ready for use. The protocol was tested with varying pressures, drying times, volume amounts, and preparation methods to determine if highquality needles can be produced. A MN is a method of DNA extraction where the patch is applied to a leaf, the needles penetrate the leaf, breaking the rigid plant cell wall to isolate the DNA. A protocol for this method of extraction was tested to ensure the patch could produce the needed yield and purity. The tests varied by the number of patches, number of applications, and plant type. To automate the MN extraction method, motion control will utilize two separate axis tables which move in the x and y directions. The y-axis table will have an end effector that fits a MN patch and will have the ability to apply the patch to the leaf sample. This end effector will also act as a lid for a downstream processing module. The other axis will position the leaf sample and processing container so that the patch can be applied accurately. The Joint Comprehensive Sequencing System (JCSS) module integrates all the components together. The output of this module feeds into the NASA Charged Information-Storage Polymer Preparation System (CHIPPS) for genomic sequencing. The extraction module operates using a series of syringes and tubing to pump the varying reagents needed for the extraction protocol. The results of the study proved that MN patches are a viable method of DNA extraction. While fabrication of high-quality needles was unsuccessful, the protocol was able to be further developed using centrifugation. The integrated design between the motion control and the JCSS enabled the potential for automation with a complete conceptual design and prototype. Future research and development for this study would include (1) further testing for fabrication (2) expanding the range of plant species compatible with the MN patch, and (3) building a working prototype for the integrated system.

Peter Ling↗

NASA NTRS1994

Microbial biofilm formation and its consequences for the CELSS program

A major goal of the Controlled Ecology Life Support System (CELSS) program is to provide reliable and efficient life support systems for long-duration space flights. A principal focus of the program is on the growth of higher plants in growth chambers. These crops should be grown without the risk of damage from microbial contamination. While it is unlikely that plant pathogens will pose a risk, there are serious hazards associated with microorganisms carried in the nutrient delivery systems and in the atmosphere of the growth chamber. Our experience in surface microbiology showed that colonization of surfaces with microorganisms is extremely rapid even when the inoculum is small. After initial colonization extensive biofilms accumulate on moist surfaces. These microbial films metabolize actively and slough off continuously to the air and water. During plant growth in the CELSS program, microbial biofilms have the potential to foul sensors and to plug nutrient delivery systems. In addition both metabolic products of microbial growth and degradation products of materials being considered for use as nutrient reservoirs and for delivery are likely sources of chemicals known to adversly affect plant growth.

Mitchell, R.↗