A study toward development of an automated microbial metabolism laboratory Monthly progress report
Life detection systems based on phosphate and sulfur uptake for automated microbial metabolism laboratory
LIFE DETECTOR↗
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Microbe Detector
The AutoMicrobic System (AMS) represents years of intensive research and development by McDonnell Douglas Corp. that originated with a NASA study aimed at development of a fully automated microbial detection and identification system for spacecraft use. A urine specimen is placed into the system, where it is subjected to different freeze-dried microbe nutrients for the nine most common pathogens. An electro-optical scanner studies each specimen once an hour through a 4-to-13 hour cycle, operating automatically. Changes in cell growths on each culture are monitored by computer. The presence of pathogens is indicated when growth reaches a predetermined level. The system also enumerates the pathogens and specifies the type. Developed initially to handle urine testing, AMS soon is expected to allow analyses of blood, spinal fluid, and other body fluids. An additional capability under development is "susceptibility testing," or the determination of which microbe-killing agents-such as penicillin or other antibiotics-would be most effective in eliminating the pathogens. The whole process of detecting, identifying, and enumerating the pathogens and determining susceptibility is accomplished in less half the time required for the manual procedure. The AMS minimizes human error, reduces technician time , and increases laboratory output.
Source record↗
DirectINJECT: Dosing Systems for Concentrated Liquid Biocides
Reliable and mass-efficient techniques for biocide dosing are necessary to enable residual microbial control in spacecraft potable water systems. DirectINJECT is an approach to provide residual microbial control by injection of concentrated aqueous biocide solution (e.g., highly soluble silver salt), and associated dosing systems technology developments. This would allow for well-controlled liquid biocide injection that requires low consumable mass and is insensitive to the chemistry of the target water. With a 40 g Ag+/l solution, required dosing rates would be ~1 μl/minute during water processing (for a system similar to the ISS WPA), requiring only approximately 20 ml/crew-year of concentrate. We identified and characterized the performance of three dosing systems: pressure-driven flow through a robust micro-capillary tube, a miniature peristaltic pump, and a multi-piston pump with integrated valving. We began to address several potential system reliability concerns including prevention or segregation of gas bubbles in the concentrate reservoirs as well as long-term materials compatibility and mechanical life.
Lance Dean Delzeit↗
Material Compatibility Study of Coated Metals to Maintain Biocidal Silver in a Spacecraft Potable Water System
Ionic silver-based biocide remains an option of choice for microbial control in spacecraft potable water systems. However, depletion of silver ions by wetted materials readily reduces ionic silver to nonbiocidal levels. This phenomenon occurs via various transport mechanisms at the liquid-solid interfaces between the silver-containing water and adjacent surfaces. Consequently, challenges remain in the design and/or selection of materials that can be used for the wetted system components that will help ensure silver is maintained at biocidal levels. Since silver has been baselined, and/or remains, as a potable water biocide option for several future spacecraft potable water systems, a multitiered approach is being investigated, looking at heritage materials, new processes, and/or alterative materials. This study conducted the application of eleven chemically-resistant coatings onto coupons that were cut out from three types of spacecraft-grade metals (Titanium Grade 2, Inconel 718, and 316L Stainless Steel) to investigate the performance of this material strategy as a barrier against silver depletion. In this investigation, the coated metals were immersed in a static 400 parts per billion silver biocide solution at a surface-area-to-volume ratio of 2.0 cm^(-1) and left to soak for predefined time periods. The concentration of the solution in contact with the coated samples was analyzed at various points via inductively coupled plasma mass spectrometry, and the measurements were interpreted as silver retention percentages over time. This paper summarizes the coating selection process, the configuration of the test, and the performance of each coating-metal combination at diminishing silver depletion. The preliminary results demonstrate that some of the selected coatings succeeded at maintaining biocidal silver concentrations far beyond the corresponding base metal in a similar test. The knowledge acquired through this investigation supports the deploying of coatings on plumbing lines for the integration of biocidal silver in the water system architecture of a spacecraft. Nevertheless, further testing must be continued to assess the performance of these coatings on other wetted geometries and any implications of unifying this material strategy with traditional and developmental engineering design processes for spacecraft potable water systems.
water↗
Effects of drying on nitrification activity in zeoponic medium used for long-term space missions
One component of a proposed life support system is the use of zeoponic substrates, which slowly release NH4+ into "soil" solution, for the production of plants. Nitrifying bacteria that convert NH4+ to NO3- are among the important microbial components of these systems. Survival of nitrifying bacteria in dry zeoponic substrates is needed, because the substrate would likely be stored in an air-dry state between croppings. Substrate was enriched for nitrifying bacteria and allowed to air-dry in a laminar flow hood. Stored substrate was analyzed for nitrifier survivability by measuring nitrifier activity at the beginning, 3 days, 1, 2, and 3 weeks. After rewetting, activity was approximately 9 micrograms N g-1 h-1 regardless of storage time. Nitrification rates did not decrease during storage. It seems unlikely that drying between plantings would result in practical reductions in nitrification, and reinoculation with nitrifying bacteria would not be necessary.
Non-NASA Center↗
Biofilm formation and control in a simulated spacecraft water system - Two-year results
The ability of iodine to maintain microbial water quality in a simulated spacecraft water system is being studied. An iodine level of about 2.0 mg/L is maintained by passing ultrapure influent water through an iodinated ion exchange resin. Six liters are withdrawn daily and the chemical and microbial quality of the water is monitored regularly. Stainless steel coupons used to monitor biofilm formation are being analyzed by culture methods, epifluorescence microscopy, and scanning electron microscopy. Results from the first two years of operation show a single episode of high bacterial colony counts in the iodinated system. This growth was apparently controlled by replacing the iodinated ion exchange resin. Scanning electron microscopy indicates that the iodine has limited but not completely eliminated the formation of biofilm during the first two years of operation. Significant microbial contamination has been present continuously in a parallel noniodinated system since the third week of operation.
Schultz, John R.↗
Update on Feasibility of UV LEDs in a Spacecraft Wastewater Application
As the National Aeronautics and Space Administration (NASA) expands its scope and begins to venture into long-duration, manned space flights, the function and maintenance of spacecraft water systems becomes increasingly critical and difficult. New mission requirements limit opportunities for resupply and demand extended periods of uncrewed operations. Based on lessons learned from the International Space Station (ISS), one particular challenge of future spacecraft water systems will be maintaining adequate microbial control, especially in water subsystems and component-level elements, where effective long-duration biocontrol strategies do not currently exist. To ensure the reliability and redundancy in these systems, innovative technologies will be needed to ensure mission success. After proving feasibility of commercial-off-the-shelf (COTS) devices in flow-through applications in 2018, work focused on utilizing ultraviolet (UV) light emitting diodes (LEDs) in a wastewater tank application. This paper gives an update on the tank testing that has been performed to date. Using this technology may reduce the need for consumable resupply, such as filters or biocides, and minimize crew time to make the repairs on exhausted and/or compromised systems. The ultimate rationale behind developing a UV disinfection system is to increase the stability of water systems as requirements for sterility and microbial control become more stringent for deep space missions. The resulting data from this study will be used to narrow down possible technology demonstrations for selected ISS locations to assess the use of UV technology on future exploration-class spacecraft systems.
Susan N. Gilbert↗
Post-Flight Microbial Analysis of Samples from the International Space Station Water Recovery System and Oxygen Generation System
The Regenerative, Environmental Control and Life Support System (ECLSS) on the International Space Station (ISS) includes the the Water Recovery System (WRS) and the Oxygen Generation System (OGS). The WRS consists of a Urine Processor Assembly (UPA) and Water Processor Assembly (WPA). This report describes microbial characterization of wastewater and surface samples collected from the WRS and OGS subsystems, returned to KSC, JSC, and MSFC on consecutive shuttle flights (STS-129 and STS-130) in 2009-10. STS-129 returned two filters that contained fluid samples from the WPA Waste Tank Orbital Recovery Unit (ORU), one from the waste tank and the other from the ISS humidity condensate. Direct count by microscopic enumeration revealed 8.38 x 104 cells per mL in the humidity condensate sample, but none of those cells were recoverable on solid agar media. In contrast, 3.32 x lOs cells per mL were measured from a surface swab of the WRS waste tank, including viable bacteria and fungi recovered after S12 days of incubation on solid agar media. Based on rDNA sequencing and phenotypic characterization, a fungus recovered from the filter was determined to be Lecythophora mutabilis. The bacterial isolate was identified by rDNA sequence data to be Methylobacterium radiotolerans. Additional UPA subsystem samples were returned on STS-130 for analysis. Both liquid and solid samples were collected from the Russian urine container (EDV), Distillation Assembly (DA) and Recycle Filter Tank Assembly (RFTA) for post-flight analysis. The bacterium Pseudomonas aeruginosa and fungus Chaetomium brasiliense were isolated from the EDV samples. No viable bacteria or fungi were recovered from RFTA brine samples (N= 6), but multiple samples (N = 11) from the DA and RFTA were found to contain fungal and bacterial cells. Many recovered cells have been identified to genus by rDNA sequencing and carbon source utilization profiling (BiOLOG Gen III). The presence of viable bacteria and fungi from WRS and OGS subsystems demonstrates the need for continued monitoring of ECLSS during future ISS operations and investigation of advanced antimicrobial controls.
Birmele, Michele N.↗
Submarine hydrothermal environments as sites for the origin and evolution of life
That life formed and evolved in hydrothermal environments is proposed. This hypothesis is plausible in terms of the tectonic, paleontological, and degassing history of the Earth. Submarine hydrothermal vents are the only contemporary geological environment which may truly be called primeval and which today continue to be a major source of gases and dissolved elements to the ocean. The microbial assemblages in present day hydrothermal systems therefore could be living analogues of the earliest microbial communities to develop on Earth. The evidence for the hypothesis is reviewed.
Hoffman, S.↗
Spaceflight Autonomous Multigenerational Microbial Sequencer (SAMMS) in Support of Plant-Growth Systems
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Cubesat↗
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↗
Biofilm formation and control in a simulated spacecraft water system - Interim results
The ability of iodine to control microbial contamination and biofilm formation in spacecraft water distribution systems is studied using two stainless steel water subsystems. One subsystem has an iodine level of 2.5 mg/L maintained by an iodinated ion-exchange resin. The other subsystem has no iodine added. Stainless steel coupons are removed from each system to monitor biofilm formation. Results from the first six months of operation indicate that 2.5 mg/L of iodine has limited the number of viable bacteria that can be recovered from the iodinated subsystem. Epifluorescence microscopy of the coupons taken from this subsystem, however, indicates some evidence of microbial colonization after 15 weeks of operation. Numerous bacteria have been continually removed from both the water samples and the coupons taken from the noniodinated subsystem after only 3 weeks of operation.
Schultz, John R.↗
Biogeochemical Processes in Microbial Ecosystems
The hierarchical organization of microbial ecosystems determines process rates that shape Earth's environment, create the biomarker sedimentary and atmospheric signatures of life and define the stage upon which major evolutionary events occurred. In order to understand how microorganisms have shaped the global environment of Earth and potentially, other worlds, we must develop an experimental paradigm that links biogeochemical processes with ever-changing temporal and spatial distributions of microbial population, and their metabolic properties. Photosynthetic microbial mats offer an opportunity to define holistic functionality at the millimeter scale. At the same time, their Biogeochemistry contributes to environmental processes on a planetary scale. These mats are possibly direct descendents of the most ancient biological communities; communities in which oxygenic photosynthesis might have been invented. Mats provide one of the best natural systems to study how microbial populations associate to control dynamic biogeochemical gradients. These are self-sustaining, complete ecosystems in which light energy absorbed over a diel (24 hour) cycle drives the synthesis of spatially-organized, diverse biomass. Tightly-coupled microorganisms in the mat have specialized metabolisms that catalyze transformations of carbon, nitrogen. sulfur, and a host of other elements.
DesMarais, David J.↗
BIOGEOCHEMICAL STUDIES OF PHOTOSYNTHETIC MICROBIAL MATS AND THEIR BIOTA
Photosynthetic microbial mats offer an opportunity to define holistic functionality at the millimeter scale. At the same time. their biogeochemistry contributes to environmental processes on a planetary scale. These mats are possibly direct descendents of the most ancient biological communities; communities in which oxygenic photosynthesis might have been invented. Mats provide one of the best natural systems to study how microbial populations associate to control dynamic biogeochemical gradients. These are self- sustaining, complete ecosystems in which light energy absorbed over a dial (24 hour) cycle drives the synthesis of spatially-organized, diverse biomass. Tightly-coupled microorganisms in the mat have specialized metabolisms that catalyze transformations of carbon, nitrogen, sulfur, and a host of other elements.
DesMarais, David↗
Testing of the ISS’s Charcoal HEPA Integrated Particle Scrubber (CHIPS) Filters to Ascertain an Efficiency Curve and Most Penetrating Particle Size
High air quality is a top priority in hermetically sealed environments, such as the International Space Station (ISS). Aerosol particulate filtration on the ISS is achieved via 21 HEPA filters. Traditional HEPA filters have an efficiency of 99.97%, but HEPA filters aboard the ISS have been found to achieve 99.99% efficiency. This may be due to the flight-grade HEPA filters chosen or, less likely, that they are combined with activated charcoal filters for siloxane removal in a combination referred to as Charcoal HEPA Integrated Particle Scrubbers (CHIPS) filters. All filters have a Most Penetrating Particle Size (MPPS), but, until now, the MPPS and efficiency curve of the CHIPS filters was not known. The efficiency curve of the CHIPS filters was ascertained at NASA Glenn Research Center through standard salt crystal aerosolization. This method was selected over aerosolized oil droplets due to a desire to preserve the filter’s function for further testing. The CHIPS filters were tested as they are oriented on the ISS (activated charcoal filter followed by a flight-grade HEPA filter), with the HEPA portion alone, and with the activated charcoal portion alone. The results of this experiment are generally useful but can be especially useful for the microbiology and Environmental Control and Life Support Systems (ECLSS). This is due to microbial contamination being a potential concern in the air revitalization systems on the ISS. Knowing the MMPS of the CHIPS filters, in conjunction with levels of airborne microbes, is useful for predicting which and how many microbes are likely to get through the HEPA filters that sit in the beginning of the ISS airstream. It may also be useful to other groups at NASA to know how many particulates are able to pass through high-efficiency filtration, and even for healthcare cleanrooms and other air purification pursuits.
A Nicolas Whitlock↗
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 crop 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 growing systems in space habitats. Microorganisms transported into space will most likely occur as 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, will also be presented.
Gonzales, A. A.↗
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↗
Corrosion consequences of microfouling in water reclamation systems
This paper examines the potential fouling and corrosion problems associated with microbial film formation throughout the water reclamation system (WRS) designed for the Space Station Freedom. It is shown that the use of advanced metal sputtering techiques combined with image analysis and FTIR spectroscopy will present realistic solutions for investigating the formation and function of biofilm on different alloys, the subsequent corrosion, and the efficiency of different treatments. These techniques, used in combination with electrochemical measurements of corrosion, will provide a powerful approach to examinations of materials considered for use in the WRS.
Ford, Tim↗