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All sourcesNASA NTRSDOE OSTI
NASA NTRS2016

Hollow Fiber Membrane Bioreactor Systems for Wastewater Processing: Effects of Environmental Stresses Including Dormancy Cycling and Antibiotic Dosing

Membrane-aerated biofilm reactors (MABRs) have been studied for a number of years as an alternate approach for treating wastewater streams during space exploration. While the technology provides a promising pre-treatment for lowering organic carbon and nitrogen content without the need for harsh stabilization chemicals, several challenges must be addressed before adoption of the technology in future missions. One challenge is the transportation of bioreactors containing intact, active biofilms as a means for rapid start-up on the International Space Station or beyond. Similarly, there could be a need for placing these biological systems into a dormant state for extended periods when the system is not in use, along with the ability for rapid restart. Previous studies indicated that there was little influence of storage condition (4 or 25 C, with or without bulk fluid) on recovery of bioreactors with immature biofilms (48 days old), but that an extensive recovery time was required (20+ days). Bioreactors with fully established biofilms (13 months) were able to recover from a 7-month dormancy within 4 days (approximately 1 residence). Further dormancy and recovery testing is presented here that examines the role of biofilm age on recovery requirements, repeated dormancy cycle capabilities, and effects of long-duration dormancy cycles (8-9 months) on HFMB systems. Another challenge that must be addressed is the possibility of antibiotics entering the wastewater stream. Currently, for most laboratory tests of biological water processors, donors providing urine may not contribute to the study when taking antibiotics because the effects on the system are yet uncharacterized. A simulated urinary tract infection event, where an opportunistic, pathogenic organism, E. coli, was introduced to the HFMBs followed by dosing with an antibiotic, ciprofloxacin, was completed to study the effect of the antibiotic on reactor performance and to also examine the development of antibiotic-resistant communities within the system.

Bioreactor Systems↗
NASA NTRSDate not supplied

Characterization of Microbe Resistant Coatings for Use in the ISS Water System

The ISS is a highly controlled environment; however, there are still microbes on it that astronauts bring from their own microbiome. Once within the ISS water treatment system, these microbes not only pose a health risk for astronauts, but they can form biofilms, causing material degradation and system failure. Due to their protective structure, biofilms are notoriously resistant to disinfectants and antibiotics. One proposed solution is applying polymer coatings to the metal parts of the water system to prevent biofilms from forming. Two types of polyampholyte polymers with previously demonstrated nonfouling properties were sent to the ISS in 2021 to test their viability for preventing microbial adhesion in space. However, the only microbial assays performed were microscopy. To enhance this ongoing research project, I developed procedures for culturing, applying, and quantifying Ralstonia pickettii on these polymers. R. pickettii was chosen because it is one of the most prevalent microbes found in the ISS water treatment system. Optical density data was recorded to obtain a growth curve demonstrating a doubling time between 2.83 and 4.88 hours in 30°C, not shaken conditions. Optical density was also correlated to colony forming units in a plating experiment. Another result was design (and pilot testing) of a surface colonization experiment. These results provided vital knowledge about R. pickettii, which will be used for a 2023 payload of antimicrobial polymer coatings on stainless steel, and improved methods for biofilm analysis upon its return to evaluate the efficacy of the coatings.

microbiology↗
NASA NTRSDate not supplied

CDC Bioreactor Configuration Method for Volume Level Control with Controlled Inlet and Outlet Flow

Environmental Control Life Support Systems and other microbiological biofilm studies often utilize small scale bioreactors. Among these options are the popular BioSurface CDC bioreactors, currently being utilized by Marshall Space Flight Center groups researching the impact of biofilms on life support systems. After a recent experimental regime, it was determined additional equipment could be added to augment the capabilities of the bioreactors. Previous research configurations such as ASTM E3161 – 18 relied on an outlet stream gravity draining from a side drain port located above sampling coupons. This limited applications to experimental conditions with a controlled inlet and uncontrolled outlet flow. With the introduction of a small single pole single throw (SPST) reed float switch, a peristaltic pump connected to the outlet drain and a chassis controller, the bioreactor is able to maintain a set level. The modification allows additional variables to be tested, including highly adjustable fill and flush cycles, bioreactor volume, draining and filling control. Once the configuration modifications were implemented through the installation of the new equipment, data was collected to ensure the stability of the level measurements. As the level control switch is a float switch, consideration was taken into account for effects of internal stirring speed, along with effects of inlet and outlet flow rate. Data presented in this study will illustrate the stability and effectiveness of the configuration changes in equipment made to the bioreactor. These configuration changes are proving an effective control method to conduct biofilm mitigation techniques for ECLSS hardware research. The controlled level capabilities allow for a constant drip feed flow rate into the bioreactors, a key aspect of the biofilm mitigation testing.

CDC Bioreactor↗
NASA NTRSDate not supplied

CDC Bioreactor Configuration Method for Volume Level Control with Controlled Inlet and Outlet Flow

Environmental Control Life Support Systems and other microbiological biofilm studies often utilize small scale bioreactors. Among these options are the popular BioSurface CDC bioreactors, currently being utilized by Marshall Space Flight Center groups researching the impact of biofilms on life support systems. After a recent experimental regime, it was determined additional equipment could be added to augment the capabilities of the bioreactors. Previous research configurations such as ASTM E3161-21 relied on an outlet stream gravity draining from a side drain port located above sampling coupons. This limited applications to experimental conditions with a controlled inlet and uncontrolled outlet flow. With the introduction of a small single pole single throw (SPST) reed float switch and a peristaltic pump connected to the outlet drain and a chassis controller, the bioreactor is able to maintain a set level. The modification allows additional variables to be tested, including highly adjustable fill and flush cycles, bioreactor volume, draining and filling control. Once the configuration modifications were implemented through the installation of the new equipment, data was collected to ensure the stability of the level measurements. As the level control switch is a float switch, consideration was taken into account for effects of internal stirring speed, along with effects of inlet and outlet flow rate. Data presented in this study will illustrate the stability and effectiveness of the configuration changes in equipment made to the bioreactor. These configuration changes have demonstrated control method to conduct biofilm mitigation techniques for ECLSS hardware research. The controlled level capabilities allow for a constant drip feed flow rate into the bioreactors, a key aspect of the biofilm mitigation testing.

CDC Bioreactor↗
NASA NTRSDate not supplied

CDC Bioreactor Configuration Method for Volume Level Control with Controlled Inlet and Outlet Flow

Environmental Control Life Support Systems and other microbiological biofilm studies often utilize small scale bioreactors. Among these options are the popular BioSurface CDC bioreactors, currently being utilized by Marshall Space Flight Center groups researching the impact of biofilms on life support systems. After a recent experimental regime, it was determined additional equipment could be added to augment the capabilities of the bioreactors. Previous research configurations such as ASTM E3161-21 relied on an outlet stream gravity draining from a side drain port located above sampling coupons. This limited applications to experimental conditions with a controlled inlet and uncontrolled outlet flow. With the introduction of a small single pole single throw (SPST) reed float switch and a peristaltic pump connected to the outlet drain and a chassis controller, the bioreactor is able to maintain a set level. The modification allows additional variables to be tested, including highly adjustable fill and flush cycles, bioreactor volume, draining and filling control. Once the configuration modifications were implemented through the installation of the new equipment, data was collected to ensure the stability of the level measurements. As the level control switch is a float switch, consideration was taken into account for effects of internal stirring speed, along with effects of inlet and outlet flow rate. Data presented in this study will illustrate the stability and effectiveness of the configuration changes in equipment made to the bioreactor. These configuration changes have demonstrated control method to conduct biofilm mitigation techniques for ECLSS hardware research. The controlled level capabilities allow for a constant drip feed flow rate into the bioreactors, a key aspect of the biofilm mitigation testing.

CDC Bioreactor↗
NASA NTRSDate not supplied

CDC Bioreactor Configuration Method for Volume Level Control with Controlled Inlet and Outlet Flow

Environmental Control Life Support Systems and other microbiological biofilm studies often utilize small scale bioreactors. Among these options are the popular BioSurface CDC bioreactors, currently being utilized by Marshall Space Flight Center groups researching the impact of biofilms on life support systems. After a recent experimental regime, it was determined additional equipment could be added to augment the capabilities of the bioreactors. Previous research configurations such as ASTM E3161-21 relied on an outlet stream gravity draining from a side drain port located above sampling coupons. This limited applications to experimental conditions with a controlled inlet and uncontrolled outlet flow. With the introduction of a small single pole single throw (SPST) reed float switch and a peristaltic pump connected to the outlet drain and a chassis controller, the bioreactor is able to maintain a set level. The modification allows additional variables to be tested, including highly adjustable fill and flush cycles, bioreactor volume, draining and filling control. Once the configuration modifications were implemented through the installation of the new equipment, data was collected to ensure the stability of the level measurements. As the level control switch is a float switch, consideration was taken into account for effects of internal stirring speed, along with effects of inlet and outlet flow rate. Data presented in this study will illustrate the stability and effectiveness of the configuration changes in equipment made to the bioreactor. These configuration changes have demonstrated control method to conduct biofilm mitigation techniques for ECLSS hardware research. The controlled level capabilities allow for a constant drip feed flow rate into the bioreactors, a key aspect of the biofilm mitigation testing.

CDC Bioreactor↗
NASA NTRS1994

Continuous monitoring of bacterial attachment

A major concern with the Space Station Freedom (SSF) water supply system is the control of longterm microbial contamination and biofilm development in the water storage and distribution systems. These biofilms have the potential for harboring pathogens as well as microbial strains containing resistance factors that could negatively influence crew health. The proposed means for disinfecting the water system on SSF (iodine) may encourage the selection of resistant strains. In fact, biofilm bacteria were observed in water lines from the Space Shuttle Columbia (OV-102); therefore, an alternative remediation method is required to disinfect spacecraft water lines. A thorough understanding of colonization events and the physiological parameters that will influence bacteria adhesion is required. The limiting factor for development of this technology is the ability to continuously monitor adhesion events and the effects of biocides on sessile bacteria. Methods were developed to allow bacterial adhesion and subsequent biocidal treatment to be monitored continuously. This technique couples automated image analysis with a continuous flow of a bacterial suspension through an optical flow cell. A strain of Pseudomonas cepacia isolated from the water supply of the Space Shuttle Discovery (OV-103) during STS-39 was grown in a nitrogen-limited continuous culture. This culture was challenged continuously with iodine during growth, and the adhesion characteristics of this strain was measure with regard to flow rate. Various biocides (ozone, hypochlorite, and iodine) were added to the flow stream to evaluate how well each chemical removed the bacteria. After biocide treatment, a fresh bacterial suspension was introduced into the flow cell, and the attachment rate was evaluated on the previously treated surface. This secondary fouling was again treated with biocide to determine the efficacy of multiple batch chemical treatments in removing biofilm.

Koeing, D. W.↗
NASA NTRS2012

Engineered Multifunctional Surfaces for Fluid Handling

Designs incorporating variations in capillary geometry and hydrophilic and/or antibacterial surface properties have been developed that are capable of passive gas/liquid separation and passive water flow. These designs can incorporate capillary grooves and/or surfaces arranged to create linear and circumferential capillary geometry at the micro and macro scale, radial fin configurations, micro holes and patterns, and combinations of the above. The antibacterial property of this design inhibits the growth of bacteria or the development of biofilm. The hydrophilic property reduces the water contact angle with a treated substrate such that water spreads into a thin layer atop the treated surface. These antibacterial and hydrophilic properties applied to a thermally conductive surface, combined with capillary geometry, create a novel heat exchanger capable of condensing water from a humid, two-phase water and gas flow onto the treated heat exchanger surfaces, and passively separating the condensed water from the gas flow in a reduced gravity application. The overall process to generate the antibacterial and hydrophilic properties includes multiple steps to generate the two different surface properties, and can be divided into two major steps. Step 1 uses a magnetron-based sputtering technique to implant the silver atoms into the base material. A layer of silver is built up on top of the base material. Completion of this step provides the antibacterial property. Step 2 uses a cold-plasma technique to generate the hydrophilic surface property on top of the silver layer generated in Step 1. Completion of this step provides the hydrophilic property in addition to the antibacterial property. Thermally conductive materials are fabricated and then treated to create the antibacterial and hydrophilic surface properties. The individual parts are assembled to create a condensing heat exchanger with antibacterial and hydrophilic surface properties and capillary geometry, which is capable of passive phase separation in a reduced gravity application. The plasma processes for creating antibacterial and hydrophilic surface properties are suitable for applications where water is present on an exposed surface for an extended time, such that bacteria or biofilms could form, and where there is a need to manage the water on the surface. The processes are also suitable for applications where only the hydrophilic property is needed. In particular, the processes are applicable to condensing heat exchangers (CHXs), which benefit from the antibacterial properties as well as the hydrophilic properties. Water condensing onto the control surfaces of the CHX will provide the moist conditions necessary for the growth of bacteria and the formation of biofilms. The antibacterial properties of the base layer (silver) will mitigate and prevent the growth of bacteria and formation of biofilms that would otherwise reduce the CHX performance. In addition, the hydrophilic properties reduce the water contact angle and prevent water droplets from bridging between control surfaces. Overall, the hydrophilic properties reduce the pressure drop across the CHX.

Thomas, Chris↗
NASA NTRS2013

Fiber Treatment Effects on Bioreactor Bulk Fluid Trends

In order to facilitate the exploration of worlds beyond the borders of our planet, it is necessary to maintain sustainable levels of clean water. The remediation of water via Membrane Aerated Bioreactors (MABRs) is one such method, and the focus of this study. MARRs rely on healthy biofilms grown on hollow fiber membranes to clean non-potable water. These biofilms can take weeks to months to establish. Therefore, various fiber treatments and two inoculums were evaluated for their effect on rapid biofilm formation. Fiber treatments are as follows: sanding of the fibers with 1500 and 8000 grit sandpaper, immersion of the fibers in a 1% hydrofluoric acid solution for 12 seconds and 15 minutes, and the immersion of the fibers in a Fluoroetch® solution for 18 seconds and 5 minutes. The two inoculums utilized were sourced from healthy, established MARRs; Texas Tech University (TTU) MABR "TRL5" and Kennedy Space Center (KSC) MABR "R3". Data attained from direct bacterial cell counts of the reactor bulk fluids via fluorescent microscopy, suggests that the fluoroetching treatment combined with the TTU inoculum show the greatest biofilm creation.

Ellis, Ronald II↗
NASA NTRS2001

Signature lipids and stable carbon isotope analyses of Octopus Spring hyperthermophilic communities compared with those of Aquificales representatives

The molecular and isotopic compositions of lipid biomarkers of cultured Aquificales genera have been used to study the community and trophic structure of the hyperthermophilic pink streamers and vent biofilm from Octopus Spring. Thermocrinis ruber, Thermocrinis sp. strain HI 11/12, Hydrogenobacter thermophilus TK-6, Aquifex pyrophilus, and Aquifex aeolicus all contained glycerol-ether phospholipids as well as acyl glycerides. The n-C(20:1) and cy-C(21) fatty acids dominated all of the Aquificales, while the alkyl glycerol ethers were mainly C(18:0). These Aquificales biomarkers were major constituents of the lipid extracts of two Octopus Spring samples, a biofilm associated with the siliceous vent walls, and the well-known pink streamer community (PSC). Both the biofilm and the PSC contained mono- and dialkyl glycerol ethers in which C(18) and C(20) alkyl groups were prevalent. Phospholipid fatty acids included both the Aquificales n-C(20:1) and cy-C(21), plus a series of iso-branched fatty acids (i-C(15:0) to i-C(21:0)), indicating an additional bacterial component. Biomass and lipids from the PSC were depleted in (13)C relative to source water CO(2) by 10.9 and 17.2 per thousand, respectively. The C(20-21) fatty acids of the PSC were less depleted than the iso-branched fatty acids, 18.4 and 22.6 per thousand, respectively. The biomass of T. ruber grown on CO(2) was depleted in (13)C by only 3.3 per thousand relative to C source. In contrast, biomass was depleted by 19.7 per thousand when formate was the C source. Independent of carbon source, T. ruber lipids were heavier than biomass (+1.3 per thousand). The depletion in the C(20-21) fatty acids from the PSC indicates that Thermocrinis biomass must be similarly depleted and too light to be explained by growth on CO(2). Accordingly, Thermocrinis in the PSC is likely to have utilized formate, presumably generated in the spring source region.

Carbon Isotopes/analysis↗
NASA NTRSDate not supplied

Slippery Lubricant-Infused Silica Nanoparticulate Film Processing for Anti-Biofouling Applications

Microbial biofilm build-up in water distribution systems can pose a risk to human health and pipe material integrity. The impact is more devastating in space stations and to astronauts due to the isolation from necessary replacement parts and medical resources. As a result, there is a need for coatings to be implemented onto the inner region of the pipe to minimize the adherence and growth of biofilms. Lubricant-infused surfaces has been one such interesting material for anti-biofouling applications in which their slippery property promotes repellence to many liquids and thus prevents bacterial adherence. Textured and porous films are suitable substrate candidates to infuse and contain the lubricant. However, there is little investigation in utilizing a nanoporous thin film as the substrate material for lubricant infusion. A nanoporous film has high porosity within the structure which can promote greater lubricant infusion and retention. The implementation as a thin film structure aids to reduce material consumption and cost. In our study, we utilized a well-studied nanoporous thin film fabricated via layer-by-layer assembly of polycations and colloid silica and calcination for greater stability. The film was further functionalized to promote fluorinated groups upon the surface and pores of the film, which then could interact with a fluorinated lubricant. The pristine nanoporous film was characterized to determine its morphology, thickness, wettability, and porosity. The lubricant-infused film was then tested for its lubricant layer stability upon various washing conditions and its performance against bacterial biofilm adherence as a result of its slippery and thus anti-biofouling property.

biofilm↗
NASA NTRSDate not supplied

Analysis of CDC Bioreactor Internal Thermal Measurements and Sample Coupon Temperatures

The Center for Disease Control and Prevention (CDC) bioreactor is an integral laboratory tool for the Environmental Control and Life Support Systems (ECLSS) biofilm formation and growth research program. Critical to this research is the need to adjust and maintain various surface temperatures of the coupons housed within the CDC Bioreactor. The purpose of this study was to provide quantitative temperature gradient information when the CDC Bioreactor was operating according to several process scenarios. Two primary process parameters were evaluated. For the first set of test parameters, the liquid level was maintained at 350 mL, with an inlet flowrate of 0.1 mL/min, 1 mL/min, 10 mL/min. The liquid was allowed to gravity drain out of the outlet spout. For the second set of test parameters, the liquid level within the reactor was maintained at 550 mL, with an inlet flow of 0.1 mL/min, 1 mL/min, 10 mL/min and draining intermittently controlled to 0.8 mL/min to maintain the 550 mL level. Due to the placement of the thermocouple in the reactor, a difference in temperature occurred between the coupon surfaces and target Bioreactor temperature when operated according to the first set of test parameters. When the reactor was operated according to the second set of parameters, which resulted in the thermocouple being submerged, the temperature gradient was eliminated. The results demonstrated minimal temperature gradient between the top and bottom coupon surfaces for coupons placed in a single rod within the CDC Bioreactor for both sets of test parameters evaluated. The collection of this information helped to explain previous ECLSS biofilm formation test runs, along with providing guidance on best operating practices for future ECLSS experiments. The placement of the thermocouple also helps to explain the challenge of achieving and maintaining bulk liquid temperatures when biofilm is grown according to the standardized test methods.

CDC Bioreactor↗
NASA NTRS1992

Biodeterioration of materials in water reclamation systems

The chemicals produced by the microbial processes involved in the 'biofilms' which form on the surfaces of manned spacecraft water reclamation systems encompass both metals and organic poisons; both are potential hazards to astronaut health and the growth of the plants envisioned for closed-cycle life support systems. Image analysis is here shown to be a very useful technique for the study of biofilm formation on candidate water-processor materials for Space Station Freedom. The biodeterioration of materials exposed to biofilms can be swiftly evaluated by means of electrochemical impedance spectroscopy.

Ford, Tim↗
NASA NTRS2001

Composition of Hydrothermal Vent Microbial Communities as Revealed by Analyses of Signature Lipids, Stable Carbon Isotopes and Aquificales Cultures

Extremely thermophilic microbial communities associated with the siliceous vent walls and outflow channel of Octopus Spring, Yellowstone National Park, have been examined for lipid biomarkers and carbon isotopic signatures. These data were compared with that obtained from representatives of three Aquificales genera. Thermocrinis ruber. "Thermocrinis sp. HI", Hydrogenobacter thermophilus TK-6, Aquifex pyrophilus and Aquifex aeolicus all contained phospholipids composed not only of the usual ester-linked fatty acids, but also ether-linked alkyls. The fatty acids of all cultured organisms were dominated by a very distinct pattern of n-C-20:1 and cy-C-21 compounds. The alkyl glycerol ethers were present primarily as CIS() monoethers with the expection of the Aquifex spp. in which dialkyl glycerol ethers with a boarder carbon-number distribution were also present. These Aquificales biomarker lipids were the major constituents in the lipid extracts of the Octopus Spring microbial samples. Two natural samples, a microbial biofilm growing in association with deposition of amorphous silica on the vent walls at 92 C, and the well-known 'pink-streamers community' (PSC), siliceous filaments of a microbial consortia growing in the upper outflow channel at 87 C were analyzed. Both the biofilm and PSC samples contained mono and dialkyl glycerol ethers with a prevalence of C-18 and C-20 alkyls. Phospholipid fatty acids were comprised of both the characteristic Aquificales n-C-20:1 and cy-C-21, and in addition, a series of iso-branched fatty acids from i-C-15:0 to i-C-21:0, With i-C-17:0 dominant in the PSC and i-C-19:0 in the biofilm, suggesting the presence of two major bacterial groups. Bacteriohopanepolyols were absent and the minute quantities of archaeol detected showed that Archaea were only minor constituents. Carbon isotopic compositions of the PSC yielded information about community structure and likely physiology. Biomass was C-13-depleted (10.9%) relative to available CO2 from the source water inorganic carbon pool with lipids further depleted by 6.3% relative to biomass The C-20-21 Aquificales fatty acids of the PSC were somewhat heavier than the iso-branched fatty acids. The carbon isotopic signatures of lipid biomarkers were also explored using a pure culture, T ruber, previously isolated from the PSC. Cells grown on C02 with O2 and both H2 and thiosulfate as electron donors were only slightly depleted (3.3%) relative to the C-source while cells grown on formate with O2 showed a major discrimination (19.7%), possibly the result of a metabolic branch point involving the assimilation of C-formate to biomass and the dissimilation to CO2 associated with energy production. T. ruber lipids were slightly heavier than biomass (+1.3%) whether cells were grown using CO2 or formate. Fatty acids from CO2 grown T. ruber cells were a so slightly heavier (average +2.1%) than biomass. The relatively depleted PSC C-20-21 fatty acids suggest that any associated Thermocrinis biomass would also be similarly depleted and much too light to be explained by growth on CO2. The C-fractionations determined with the pure culture suggest that growth of Thermocrinis in the PSC is more likely to occur on formate, presumably generated by geothermal activity. This study points to the value of the analysis of the structural and isotopic composition of lipid blomarkers both in pure culture studies, and in establishing community structure and physiology, as a complement to genomic profiles of microbial diversity. This is especially so when the members of the microbial community are novel and difficult to cultivate in the laboratory.

Jahnke, Linda L.↗
NASA NTRS2011

Novel Materials for Prosthetic Liners

Existing materials for prosthetic liners tend to be thick and airtight, causing perspiration to accumulate inside the liner and potentially causing infection and injury that reduce quality of life. The purpose of this project was to examine the suitability of aerogel for prosthetic liner applications. Three tests were performed on several types of aerogel to assess the properties of each material. Moisture vapor permeability was tested by incubating four aerogel varieties with an artificial sweat solution at 37.0 C and less than 20% relative humidity for 24 hours. Two aerogel varieties were eliminated from the study due to difficulties in handling the material, and further testing proceeded with Pyrogel in 2.0 and 6.0 mm thicknesses. Force distribution was tested by compressing samples under a load of 4448 N at a rate of 2.5 mm/min. Biofilm formation was tested in a high-shear CDC Biofilm Reactor. Results showed that 2.0 mm Pyrogel blanket allowed 55.7 plus or minus 28.7% of an artificial sweat solution to transpire, and 35.5 plus or minus 27.8% transpired through 6.0 mm Pyrogel blanket. Samples also outperformed the load-bearing capabilities of existing liner materials. No statistically significant difference was found between the two Pyrogel thicknesses for either moisture vapor permeability or force distribution. In addition, biofilm formation results showed no change between the two Pyrogel thicknesses. The breathability and load bearing properties of aerogel make it a suitable material for application to prosthetic liners.

Ragolta, Carolina I.↗
NASA NTRS2012

Survival of B. Horneckiae Spores Under Ground-simulated Space Conditions

To prevent forward contamination and maintain the scientific integrity of future life detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated habitats, spore-forming microbes are highly resistant to various physical and chemical conditions, which include ionizing and UV radiation, desiccation and oxidative stress, and the harsh environment of outer space or planetary surfaces. Recently a radiation resistant, spore forming bacterial isolate, Bacillus horneckiae, was isolated from a clean room of the Kennedy Space Center where the Phoenix spacecraft was assembled. The exceptionally high tolerance of extreme conditions demonstrated by sporeforming bacteria highlighted the need to assess the viability of these microbes in situ (in real) space. The proposed BOSS (Biofilm Organisms Surfing Space) project aims to understand the mechanisms by which biofilm forming organisms, such as B. horneckiae, will potentially be able to withstand harsh space conditions. As previously stated, the spore producing ability of these species gives them increased survivability to harsh conditions. Some of the spores will have the protective exosporium layer artificially removed before the test to determine if the existence of this layer significantly changes the survivability during the mission. In preparation for that experiment, we analyzed spores which were exposed during a ground simulation, the EXPOSE R2 Biofilm Organisms Surfing Space (BOSS). Previous to exposure, spores were deposited onto spacecraft grade aluminum coupons in a spore suspension calculated to contain between 10(exp 7) and 10(exp 8) spores. This precursor series will be used to establish a baseline survivability function for comparison with the future flight tests during EXPOSE-R. For each coupon, a 10% polyvinyl alcohol (PVA) film was applied and peeled from the coupon to recover the spores. One hundred μl of sterile 10% PVA was applied to the surface of the coupon and allowed to dry for 1 hour at 37 C. The films were then removed using sterile scalpel and forceps and placed into a glass test tube containing 2 milliliters of sterile deionized water. The PVA film process was then repeated on each coupon one additional time to ensure recovery of the majority of spores. The second PVA film was added in the same glass tube as in the previous round. If the spores remained 100% viable, the test tubes should now contain between 5 X 10(exp 6) and 5 X 10(exp 7) spores per millimeter; however, it is expected that some loss of viability has occurred. In order to assess this loss, the number of colony forming, viable spores was counted. To count the colony forming units (CFUs), the spore containing solution was diluted in a process of 10-fold serial dilution by mixing successive solutions in a 100 microliter spore suspension to 900 microliter deionized H2O ratio. A sample dilution series revealed that 10(exp -3) and 10(exp -4) concentrations would be necessary for an accurate CFU count to be taken. For those two concentrations, a spread on a TSA plate was prepared and incubated at 32 C. For the samples exposed to UV radiation, the cell survivability was too low to establish a count from 100 microliter spread plating. Instead, no dilutions were performed and the entire 2 milliliter spore suspension was plated and incubated at 32 C. The plate's CFU counts were taken at 24 hours and 48 hours from the time of plating. At the end of the CFU counting the total surviving spores in each sample were calculated based on the number of CFUs that were observed per 100 microliters, or per 2 milliliters for the UV irradiated samples. The results of these calculations are shown in Figures 1 and 2.

cleanroom↗
NASA NTRS2016

Silver Ion Biocide Delivery System for Water Disinfection

U.S. space exploration missions have long considered returning to the Moon and exploration of Mars that challenge life support systems. For these long duration missions, there is interest in replacing the iodine water treatment system with ionic silver, a proven biocide. For long duration exploration missions, it is imperative that an effective biocide be used that prevents microbial growth, biofilm formation, and microbially induced corrosion in the water storage and distribution systems while minimizing logistical supply requirements associated with the biocide delivery system. Two biocide delivery systems have been developed that electrochemically produce silver ions for disinfecting water throughout the water storage and distribution system. One system uses a newly developed hybrid micro-filtration and ion-exchange membrane to produce an abundance of silver ions at the 1000 ppb level upstream in the water distribution system to prevent biofilm growth. This is followed by a downstream collection module that electrochemically removes these silver ions before the water is discharged. Another approach uses a membraneless reactor to produce a 1000 ppb silver ion concentration level that also has a mechanically driven electrode cleaning mechanism that removes oxide films ensuring long life operation. By maintaining a sufficiently high level of silver ions throughout the water storage and distribution system, biofilm formation is suppressed. This approach overcomes present concerns where spurious silver deposition occurs on the container and flow line surfaces thus lowering the silver ion concentration to unsatisfactory disinfection levels.

Slote, Benjamin M.↗
NASA NTRS2018

Microorganism Cultivation Platform for Human Life Support

A life support system for providing a growth medium for at least one photosynthetic micro-organism and for converting CO2 to O2, with reduced water use that is as low as about 4 percent of the corresponding amount of water normally required for conventional micro-organism growth. The system includes a liquid transport capillary channel, a mixed culture photosynthetic biofilm and a liquid transport substrate that is positioned between and contiguous to the capillary channel and the biofilm, where the liquid transport rate is adjustable by adjustment of the local humidity. Approximately uniform radiation is received by the biofilm and contributes to microorganism growth.

Berberoglu, Halil↗