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All sourcesNASA NTRSDOE OSTI
NASA NTRS1999

Sensitive detection of Escherichia coli O157:H7 in food and water by immunomagnetic separation and solid-phase laser cytometry

Rapid, direct methods are needed to assess active bacterial populations in water and foods. Our objective was to determine the efficiency of bacterial detection by immunomagnetic separation (IMS) and the compatibility of IMS with cyanoditolyl tetrazolium chloride (CTC) incubation to determine respiratory activity, using the pathogen Escherichia coli O157:H7. Counterstaining with a specific fluorescein-conjugated anti-O157 antibody (FAb) following CTC incubation was used to allow confirmation and visualization of bacteria by epifluorescence microscopy. Broth-grown E. coli O157:H7 was used to inoculate fresh ground beef (<17% fat), sterile 0.1% peptone, or water. Inoculated meat was diluted and homogenized in a stomacher and then incubated with paramagnetic beads coated with anti-O157 specific antibody. After IMS, cells with magnetic beads attached were stained with CTC and then an anti-O157 antibody-fluorescein isothiocyanate conjugate and filtered for microscopic enumeration or solid-phase laser cytometry. Enumeration by laser scanning permitted detection of ca. 10 CFU/g of ground beef or <10 CFU/ml of liquid sample. With inoculated meat, the regression results for log-transformed respiring FAb-positive counts of cells recovered on beads versus sorbitol-negative plate counts in the inoculum were as follows: intercept = 1.06, slope = 0.89, and r2 = 0. 95 (n = 13). The corresponding results for inoculated peptone were as follows: intercept = 0.67, slope = 0.88, and r2 = 0.98 (n = 24). Recovery of target bacteria on beads by the IMS-CTC-FAb method, compared with recovery by sorbitol MacConkey agar plating, yielded greater numbers (beef, 6.0 times; peptone, 3.0 times; water, 2.4 times). Thus, within 5 to 7 h, the IMS-CTC-FAb method detected greater numbers of E. coli O157 cells than were detected by plating. The results show that the IMS-CTC-FAb technique with enumeration by either fluorescence microscopy or solid-phase laser scanning cytometry gave results that compared favorably with plating following IMS.

Non-NASA Center↗
NASA NTRS2004

Extracellular mass transport considerations for space flight research concerning suspended and adherent in vitro cell cultures

Conducting biological research in space requires consideration be given to isolating appropriate control parameters. For in vitro cell cultures, numerous environmental factors can adversely affect data interpretation. A biological response attributed to microgravity can, in theory, be explicitly correlated to a specific lack of weight or gravity-driven motion occurring to, within or around a cell. Weight can be broken down to include the formation of hydrostatic gradients, structural load (stress) or physical deformation (strain). Gravitationally induced motion within or near individual cells in a fluid includes sedimentation (or buoyancy) of the cell and associated shear forces, displacement of cytoskeleton or organelles, and factors associated with intra- or extracellular mass transport. Finally, and of particular importance for cell culture experiments, the collective effects of gravity must be considered for the overall system consisting of the cells, their environment and the device in which they are contained. This does not, however, rule out other confounding variables such as launch acceleration, on orbit vibration, transient acceleration impulses or radiation, which can be isolated using onboard centrifuges or vibration isolation techniques. A framework is offered for characterizing specific cause-and-effect relationships for gravity-dependent responses as a function of the above parameters.

Non-NASA Center↗
NASA NTRS1966

Biology and the Exploration of Mars

Until recent years the origin of life and its possible occurrence elsewhere in the universe have been matters for speculation only. The rapid growth of molecular biology since 1940 has, to be sure, made it possible to discuss life's origins in far more precise and explicit terms than was possible earlier; and the subject entered a new experimental phase in the 1950's with successful abiogenic synthesis of important biochemical substances in conditions simulating the presumptive environment of the primitive Earth. But the real transformation that the subject has undergone stems from the spectacular growth of space technology in the last decade. The possibility of life's origin and occurrence on planets other than ours is no longer limited to idle speculation: it has entered the realm of the testable, of science in the strict sense. Given the rockets now available, and especially those available by 1969, it has become fully realistic to consider plans for the biological exploration of Mars. The study that this report seeks to interpret was initiated in June, 1964, by the Space Science Board of the National Academy of Sciences to examine this possibility. The working group comprised 36 people representing a broad spectrum of scientific interests: evolutionary biology, genetics, microbiology, biochemistry and molecular biology, animal physiology, soil chemistry, organic chemistry, planetary astronomy, geochemistry, and theoretical physics. The participants included some with considerable prior involvement in problems of space exploration and others with none. Advice was also sought outside the group of immediate participants on the potentialities of selected analytical methods for the experimental study of extraterrestrial life and its environment. More than 30 individuals contributed in this fashion written assessments of techniques in which they were particularly well versed. Our task was to examine the scientific foundations and merits of the proposal to undertake a biological exploration of Mars. What are the potential scientific yields? How valuable, if attained, would they be? What, in fact, is the possibility of life occurring on Mars? And of our detecting it with available and foreseeable technology? What could be achieved by further astronomical work from Earth? by Martian fly-by missions? by Martian orbiters? and Martian Landers? What payloads would we recommend for planetary missions? What timing and over-all strategy would we recommend for Martian exploration were we to consider it worthwhile at all? In brief, the over-all purpose was to recommend to. the government, through the Academy's Space Science Board, whether or not a biological exploration of Mars should be included in the nation's space program over the next few decades; and, further, to outline what that program, if any, should be.

CONFERENCE↗
NASA NTRS1994

Paleolimnology of the McMurdo Dry Valleys, Antarctica

The McMurdo Dry Valleys presently contain more than 20 permanent lakes and ponds, which vary markedly in character. All, with the exception of a hypersaline pond, have a perennial ice-cover. The dry valley lakes, and lakes in other ice-free regions of continental Antarctica, are unique on this planet in that they consistently maintain a thick year-round ice cover (2.8-6.0 m) over liquid water. The persistent ice covers minimize wind-generated currents and reduce light penetration, as well as restricting sediment deposition into a lake and the exchange of atmospheric gases between the water column and the atmosphere. From a paleolimnological perspective, the dry valley lakes offer an important record of catchment and environmental changes. These lakes are also modern-day equivalents of periglacial lakes that were common during glacial periods at temperate latitudes. The present lakes are mostly remnants of larger glacial lakes that occupied the valleys in the past, perhaps up to 4.6 Ma ago. Two of the valleys contain evidence of being filled with large glacial lakes within the last 10000 years. Repeated drying and filling events since then have left a characteristic impression on the salt profiles of some lakes creating a unique paleo-indicator within the water column. These events are also marked in the sediments by the concentration and dilution of certain chemical constituents, particularly salts, and are also corroborated by carbonate speciation and oxygen isotope analysis. Stratigraphic analysis of dry valley lake sediments is made difficult by the occurrence of an 'old carbon' reservoir creating spurious radiocarbon dates, and by the high degree of spatial variability in lake sedimentation. From a biological perspective, the lakes are relatively simple, containing various taxa of planktonic and benthic microorganisms, but no higher forms of life, which is an advantage to paleolimnologists because there is no bioturbation in the sediments. Useful biological paleo-indicators found in the sediments include cyanobacterial filament sheaths, diatom frustules and other eukaryotic algal cells, protozoan cysts, photosynthetic pigments, and minerals (e.g. carbonates) associated with microbial activity. Future work will benefit from fully characterizing the connection between the ice covers, environmental conditions, and paleo-indicators, thereby allowing refinement of inferences made concerning the paleoenvironment. New dating techniques need to be tested in this environment to overcome the problems associated with radiocarbon dating. The establishment of a detailed and focused paleolimnological campaign is proposed.

Non-NASA Center↗
NASA NTRS1991

Characterization of the corrosion resistance of biologically active solutions: The effects of anodizing and welding

An understanding of fabrication processes, metallurgy, electrochemistry, and microbiology is crucial to the resolution of microbiologically influenced corrosion (MIC) problems. The object of this effort was to use AC impedance spectroscopy to characterize the corrosion resistance of Type II anodized aluminum alloy 2219-T87 in sterile and biologically active media and to examine the corrosion resistance of 316L, alloy 2219-T87, and titanium alloy 6-4 in the welded and unwelded conditions. The latter materials were immersed in sterile and biologically active media and corrosion currents were measured using the polarization resistance (DC) technique.

Walsh, Daniel W.↗
NASA NTRS1991

Microbiology on Space Station Freedom

This panel discussion convened in Houston, Texas, at the Lunar and Planetary Institute, on November 6 to 8, 1989, to review NASA's plans for microbiology on Space Station Freedom. A panel of distinguished scientists reviewed, validated, and recommended revisions to NASA's proposed acceptability standards for air, water, and internal surfaces on board Freedom. Also reviewed were the proposed microbiology capabilities and monitoring plan, disinfection procedures, waste management, and clinical issues. In the opinion of this advisory panel, ensuring the health of the Freedom's crews requires a strong goal-oriented research effort to determine the potential effects of microorganisms on the crewmembers and on the physical environment of the station. Because there are very few data addressing the fundamental question of how microgravity influences microbial function, the panel recommended establishing a ground-based microbial model of Freedom, with subsequent evaluation using in-flight shuttle data. Sampling techniques and standards will be affected by both technological advances in microgravity-compatible instrumentation, and by changes in the microbial population over the life of the station.

Pierson, Duane L.↗
NASA NTRS2020

Silver Foam A Long-Term Passive Biocide for Potable Water Systems: 2020 Update

A spacecraft water disinfection system suitable for extended length space exploration missions should prevent or control the growth of microbes, prevent or limit biofilm formation, and prevent microbiologically-influenced corrosion. In addition, the system should have minimal maintenance requirements, the effluent should be chemically compatible with all materials in contact with the water, be safe for human consumption, and suitable to be shared across international spacecraft platforms and mission architectures. Silver ions are a proven broad spectrum biocide and the chosen potable water biocide for future exploration missions. The leading technology for actively dosing silver in future water systems is based on electrolytic production. Several challenges remain with this approach which have prompted additional investigations into alternative dosing techniques. Control-release technology is an attractive option for developing a high-reliability passive silver dosing device. This paper describes the development of a nanoparticle (NP)/polyurethane (PU) composite foam for the controlled release of silver ions, and is intended to build upon the 2019 International Conference on Environmental Systems (ICES) paper of the same name. In this paper, the technical background and results from the updated silver chloride (AgCl) NP/PU composite foam synthesis and property testing is provided. The ultimate goal of the project is to develop a stable and reliable passive dosing silver ion release device for use in future spacecraft potable water systems.

Irwin, Tesia D.↗
NASA NTRSDate not supplied

Silver Foam: A Novel Approach for Long-Term Passive Dosing of Biocide in Spacecraft Potable Water Systems

A spacecraft water disinfection system suitable for extended length space exploration should prevent or control the growth of microbes, prevent or limit biofilm formation, and prevent microbiologically influenced corrosion. In addition, the system should have minimal maintenance requirements,should be chemically compatible with all materials in contact with the water, be safe for human consumption, and suitable to be shared across international spacecraft platforms and mission architectures. Silver ions are a proven broad-spectrum biocide under consideration as a potable water biocide for future exploration missions. The leading technology for actively dosing silver in future water systems is based on electrolytic production. Several challenges remain with this approach that have prompted additional investigations into alternative dosing techniques. Control-release technology is an attractive option for developing a high-reliability passive silver dosing device. This paper describes the development of a silver chloride (AgCl) nanoparticle (NP)/polyurethane (PU) composite foam for the controlled release of silver ions, and is intended to build upon the 2019 International Conference on Environmental Systems (ICES) paper number 272.This paper provides the technical background and results from the updated AgClNP/PU composite foam (AgFoam) synthesis and property testing. The ultimate goal of the project is to develop a stable and reliable passive dosing silver ion release device for use in future spacecraft potable water systems.

Tesia Dawn Irwin↗
NASA NTRSDate not supplied

Silver Foam: A Novel Approach for Long-Term Passive Dosing of Biocide in Spacecraft Potable Water Systems

A spacecraft water disinfection system, suitable for extended length space exploration, should prevent or control the growth of microbes, prevent or limit biofilm formation, and prevent microbiologically influenced corrosion. In addition, the system should have minimal maintenance requirements, be chemically compatible with all materials in contact with the water, be safe for human consumption, and be suitable to be shared across international spacecraft platforms and mission architectures. Silver ions are a proven broad-spectrum potable water biocide under investigation for future exploration missions. The competing technology for dosing silver ions in future water systems is based on actively dosing the ions via electrolytic production. Several challenges with this approach have prompted additional investigations into alternative dosing techniques. Control-release technology is an attractive option for developing a high-reliability passive silver dosing device. This paper describes the development of a nanoparticle/polyurethane (NP/PU) composite foam for the controlled release of silver ions, and is intended to build upon the 2020 International Conference onEnvironmental Systems (ICES) paper number 128. This paper provides the technical background and performance results from the updated silver chloride (AgCl) NP/PU. The ultimate goal of the project is to develop a stable and reliable passive dosing silver ion release device for use in future spacecraft potable water systems.

Tesia D. Irwin↗
NASA NTRSDate not supplied

Silver Foam: A Novel Approach for Long-Term Passive-Dosing of Biocide in Spacecraft Potable Water Systems – Update 2022

A spacecraft water disinfection system suitable for extended length space exploration should meet a list of requirements. It should prevent or control the growth of microbes, prevent or limit biofilm formation, and prevent microbiologically influenced corrosion. In addition, the system should have minimal maintenance requirements, be chemically compatible with all materials in contact with the water, be safe for human consumption, and be suitable to be shared across international spacecraft platforms and mission architectures. Silver ions are a proven broad-spectrum potable water biocide under investigation for future exploration missions. The competing technology for dosing silver ions in future water systems is based on actively dosing the ions via electrolytic production. Several challenges with this approach have prompted additional investigations into alternative dosing techniques. Control-release technology is an attractive option for developing a high-reliability passive silver dosing device. This paper describes the continued development of a nanoparticle/polyurethane (NP/PU) composite foam for the controlled release of silver ions and is intended to build upon previously published work. This paper provides the technical background and performance results (wash testing and ongoing long-term silver ion release testing) from the silver chloride (AgCl) NP/PU composite foams. The goal of the project is to develop a stable and reliable passive-dosing silver ion release device for use in future spacecraft potable water systems.

Tesia D Irwin↗
NASA NTRSDate not supplied

Silver Foam: A Novel Approach for Long-Term Passive Dosing of Biocide in Spacecraft Potable Water Systems – Update 2023

A spacecraft water disinfection system, suitable for extended length space exploration, should prevent or control the growth of microbes, prevent or limit biofilm formation, and prevent microbiologically influenced corrosion. In addition, the system should have minimal maintenance requirements, be chemically compatible with all materials in contact with the water, be safe for human consumption, and be suitable to be shared across international spacecraft platforms and mission architectures. Silver ions are a proven broad-spectrum potable water biocide under investigation for future exploration missions. The competing technology for dosing silver ions in future water systems is based on actively dosing the ions via electrolytic production. Several challenges with this approach have prompted additional investigations into alternative dosing techniques. Control-release technology is an attractive option for developing a high-reliability passive silver dosing device. This paper describes the continued development of a nanoparticle/polyurethane (NP/PU) composite foam for the controlled release of silver ions and is intended to build upon the 2022 International Conference on Environmental Systems (ICES) paper number 97. This paper provides the technical background and performance test results (ongoing long-term silver ion release testing and product variability testing) from the silver chloride (AgCl) NP/PU composite foams. The ultimate goal of the project is to develop a stable and reliable passive dosing silver ion release device for use in future spacecraft potable water systems.

Tesia D. Irwin↗
NASA NTRSDate not supplied

Silver Foam: A Novel Approach for Long-Term Passive Dosing of Biocide in Spacecraft Potable Water Systems – Update 2024

A spacecraft water disinfection system, suitable for extended length space exploration, should prevent or control the growth of microbes, prevent or limit biofilm formation, and prevent microbiologically influenced corrosion. In addition, the system should have minimal maintenance requirements, be chemically compatible with all materials in contact with the water, be safe for human consumption, and be suitable to be shared across international spacecraft platforms and mission architectures. Silver ions are a proven broad-spectrum potable water biocide under investigation for future exploration missions. The competing technology for dosing silver ions in future water systems is based on actively dosing the ions via electrolytic production. Several challenges with this approach have prompted additional investigations into alternative dosing techniques. Control-release technology is an attractive option for developing a high-reliability passive silver dosing device. This paper describes the continued development of a nanoparticle/polyurethane (NP/PU) composite foam for the controlled release of silver ions and is intended to build upon the 2023 International Conference on Environmental Systems (ICES) paper number 251. This paper provides the technical background and performance results for the product variability testing and microbial check valve (MCV) testing of the silver chloride (AgCl) NP/PU composite foams, referred to as AgFoams. The ultimate goal of the project is to develop a stable and reliable passive dosing silver ion release device for use in future spacecraft potable water systems.

Tesia D Irwin↗
NASA NTRS2009

Microbial Risk Assessment

Historically, microbiological spaceflight requirements have been established in a subjective manner based upon expert opinion of both environmental and clinical monitoring results and the incidence of disease. The limited amount of data, especially from long-duration missions, has created very conservative requirements based primarily on the concentration of microorganisms. Periodic reevaluations of new data from later missions have allowed some relaxation of these stringent requirements. However, the requirements remain very conservative and subjective in nature, and the risk of crew illness due to infectious microorganisms is not well defined. The use of modeling techniques for microbial risk has been applied in the food and potable water industries and has exceptional potential for spaceflight applications. From a productivity standpoint, this type of modeling can (1) decrease unnecessary costs and resource usage and (2) prevent inadequate or inappropriate data for health assessment. In addition, a quantitative model has several advantages for risk management and communication. By identifying the variable components of the model and the knowledge associated with each component, this type of modeling can: (1) Systematically identify and close knowledge gaps, (2) Systematically identify acceptable and unacceptable risks, (3) Improve communication with stakeholders as to the reasons for resource use, and (4) Facilitate external scientific approval of the NASA requirements. The modeling of microbial risk involves the evaluation of several key factors including hazard identification, crew exposure assessment, dose-response assessment, and risk characterization. Many of these factors are similar to conditions found on Earth; however, the spaceflight environment is very specialized as the inhabitants live in a small, semi-closed environment that is often dependent on regenerative life support systems. To further complicate modeling efforts, microbial dose-response characteristics may be affected by a potentially dysfunctional crew immune system during a mission. In addition, microbial virulence has been shown to change under certain conditions during spaceflight, further complicating dose-response characterization. An initial study of the applicability of microbial risk assessment techniques was performed using Crew Health Care System (CHeCS) operational data from the International Space Station potable water systems. The risk of infection from potable water was selected as the flight systems and microbial ecology are well defined. This initial study confirmed the feasibility of using microbial risk assessment modeling for spaceflight systems. While no immediate threat was detected, the study identified several medically significant microorganisms that could pose a health risk if uncontrolled. The study also identified several specific knowledge gaps in making a risk assessment and noted that filling these knowledge gaps is essential as the risk estimates may change by orders of magnitude depending on the answers. The current phase of the microbial risk assessment studies focuses on the dose-response relationship of specific infectious agents, focusing on Salmonella enterica Typhimurium, Pseudomonas spp., and Escherichia coli, as their evaluation will provide a better baseline for determining the overall hazard characterization. The organisms were chosen as they either have been isolated on spacecraft or have an identified route of infection during a mission. The characterization will utilize dose-response models selected either from the peer-reviewed literature and/or by using statistical approaches. Development of these modeling and risk assessment techniques will help to optimize flight requirements and to protect the safety, health, and performance of the crew.

Ott, C. M.↗
NASA NTRS1971

Microbiologic assay of space hardware.

Review of the procedures used in the microbiological examination of space hardware. The general procedure for enumerating aerobic and anaerobic microorganisms and spores is outlined. Culture media and temperature-time cycles used for incubation are reviewed, along with assay systems designed for the enumeration of aerobic and anaerobic spores. The special problems which are discussed are involved in the precise and accurate enumeration of microorganisms on surfaces and in the neutralization of viable organisms buried inside solid materials that could be released to a planet's surface if the solid should be fractured. Special attention is given to sampling procedures including also the indirect techniques of surface assays of space hardware such as those using detachable or fallout strips. Some data on comparative levels of microbial contamination on lunar and planetary spacecraft are presented.

Favero, M. S.↗
NASA NTRS1991

Diversity of micro-fungi in an Antarctic dry valley

The fungal microflora of a dry valley in Southern Victoria Land near McMurdo Sound, Antarctica, was investigated. Samples were collected from introduced objects such as a mummified penguin and spent chewing tobacco in addition to the sparse soil found in rock fissures, isolated moss colonies, shoreline deposit materials, CaCO3 precipitates, and microbial mat debris obtained from the frozen surface of the lake in the basin of Taylor Valley. Using conventional media and techniques, all collection sites yielded populations of yeasts and filamentous fungi. Water samples and live microbial mats from beneath the lake ice yielded species of fungi along with an abundance of bacteria.

NASA Discipline Exobiology↗
NASA NTRS2017

A Ratio of Spore to Viable Organisms: A Case Study of the JPL-SAF Cleanroom

Spacecraft surfaces that are destined to land on potential life-harboring celestial bodies are required to be rigorously cleaned and continuously monitored for spore bioburden as a proxy for spacecraft cleanliness. The NASA standard assay (NSA), used for spacecraft bioburden estimates, specifically measures spores that are cultivable, aerobic, resistant to heat shock, and grow at 30 C in a nutrient-rich medium. Since the vast majority of microorganisms cannot be cultivated using the NSA, it is necessary to utilize state-of-the art molecular techniques to better understand the presence of all viable microorganisms, not just those measured with the NSA. In this study, the nutrient-deprived low biomass cleanrooms, where spacecraft are assembled, were used as a surrogate for spacecraft surfaces to measure the ratio of NSA spores in relation to the total viable microorganism population in order to make comparisons with the 2006 Space Studies Board (SSB) estimate of 1 spore per approximately 50,000 viable organisms. Ninety-eight surface wipe samples were collected from the Spacecraft Assembly Facility (SAF) cleanroom at the Jet Propulsion Laboratory (JPL) over a 6-month period. The samples were processed and analyzed using classical microbiology along with molecular methodology. Traditional microbiology plating methods were used to determine the cultivable bacterial, fungal, and spore populations. Molecular assays were used to determine the total organisms (TO, dead and live) and the viable organisms (VO, live). The TO was measured using adenosine triphosphate (ATP) and quantitative polymerase chain reaction (qPCR) assays. The VO was measured using internal ATP, propidium monoazide (PMA)-qPCR, and flow cytometry (after staining for viable microorganisms) assays. Based on the results, it was possible to establish a ratio between spore counts and VO for each viability assay. The ATP-based spore to VO ratio ranged from 149-746, and the bacterial PMA-qPCR assay-based ratio ranged from 314-1,491 VO, per spore. The most conservative estimate came from fluorescent-assisted cell sorting (FACS), which estimated the ratio to be 12,091 VO per 1 NSA spore. Since archaeal (less than 1%) and fungal (approximately 2%) populations were negligible, the spore to VO ratios were based on bacterial population estimates. The most conservative ratio from this study can be used as a replacement for the SSB estimate on nutrient-deprived (oligotrophic) desiccated spacecraft surfaces, to estimate the VO from NSA measurements without utilizing state-of-the art molecular methods that are costly and require more biomass than is typically found on spacecraft surfaces.

Hendrickson, Ryan↗
NASA NTRS2017

Comprehensive Measurement of Microbial Burden in Nutrient-Deprived Cleanrooms

Spacecraft surfaces that are destined to land on potential life-harboring celestial bodies are required to be rigorously cleaned and continuously monitored for spore bioburden as a proxy for spacecraft cleanliness. The NASA standard spore assay (NSA), used for spacecraft bioburden estimates, specifically measures spores that are cultivable, aerobic, resistant to heat shock, and grow at 30˚C in a nutrient-rich medium. Since the vast majority of microorganisms cannot be cultivated using the NSA assay, it is necessary to utilize state-of-the art molecular techniques to better understand the presence of all viable microorganisms, not just those measured with the NSA. In this study, the nutrient-deprived low biomass cleanrooms, where spacecraft are assembled, were used as a surrogate to spacecraft surfaces to measure the ratio of NSA spores in relation to the total viable microorganism population to compare with a 2006 space studies report that estimates that for every 1 spore there is approximately 50,000 viable organisms. Ninety-eight surface wipe samples were collected from the spacecraft assembly facility (SAF) cleanroom at the Jet Propulsion Laboratory (JPL) over a 6-month period. The samples were processed and analyzed using classical microbiology along with molecular assays. Traditional microbiology plating methods were used to determine the cultivable bacterial, fungal, and spore populations. Molecular assays were used to determine the total organisms (TO, dead and live) and the viable organisms (VO, live). The TO was measured using adenine triphosphate (ATP) and quantitative polymerase chain reaction (qPCR) assays. The VO was measured using internal ATP, propidium monoazide (PMA)-qPCR, and flow cytometry (after staining for viable microorganisms) assays. Based on the results, it was possible to establish a ratio between spore counts and VO for each viability assay. The ATP based spore to VO ratio ranged from 149 – 746 and the bacterial PMA-qPCR assay based ratio ranged from 314 – 1491 VO. The most conservative estimate came from FACS, which estimated the ratio to be 12,091 VO per 1 NSA spore. Since archaeal (<1%) and fungal (~2%) populations were negligible, the spore to VO ratios were based on bacterial population estimates. The most conservative ratio from this study can be used as a replacement for the SSB estimate on nutrient-deprived (oligotrophic) desiccated spacecraft surfaces, to estimate the VO from NSA measurements without utilizing state-of-the art molecular methods that are costly and require more biomass than is typically found of spacecraft surfaces.

Venkateswaran, Kasthuri↗
NASA NTRSDate not supplied

KSC to EDEN ISS: Outredgeous Lettuce Growth on the Passive Porous Tube Nutrient Delivery System (PPTNDS)

EDEN ISS is a controlled environment greenhouse established near the Neumayer Station III in Antarcticawith the mission to enable fresh crop production for the crew who are working in this remote location. EDEN provides a space to conduct plant biology and related research in a unique and isolated controlled environment (CE), while providing a platform to test developing methods relevant to long duration manned space missions. In 2021, Kennedy Space Center (KSC) scientist Jess Bunchek fulfilled a one-year mission at EDEN. During that time a variety of fresh crops were grown to record horticultural and microbiological data as well as provide the crew with fresh food to eat. One method for crop growth included an experimental irrigation system called the Passive Porous Tube Nutrient Delivery System (PPTNDS) which was developed at KSC as a microgravity-capable plant irrigation system designed to support Space Plant Biology investigations and/or crop growth during space missions. The PPTNDS was employed to grow Outredgeous Lettuce (OL) during two growouts at EDEN, (harvesting at Day After Planting (DAP) 61 and 63) and two growouts at KSC where plant growth and micro sample data were captured. This paper presents the data collected from the PPTNDS and compares it to growth on a Nutrient Film Technique (NFT) hydroponic system operated in the same environment. The result shows a system that grows OL at a reduced rate compared to the NFT system but uses less water and crew interaction to operate.

Jacob Torres↗