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Microbial film formation throughout the water reclamation systems proposed for use in the NASA Space Station Freedom poses serious corrosion risks. Choice of materials for construction of these systems must include evaluation of the potential for microbially influenced corrosion. The development of an active and therefore potentially corrosive microbial biofilm on metal surfaces is influenced by the nature of the metal substratum. This has been shown by scanning electron microscopy, isolation and identification of attached bacteria and measurements of biomass and activity. However, these techniques do not allow direct ‘ real-time’ measurement of biofilm formation and subsequent materials degradation. This is necessary to assess the efficacy of biocides and alternative remedial measures. This paper presents potential fouling and corrosion problems associated with water reclamation system design for the NASA orbiting space station.

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 Six liters are withdrawn daily and the chemical and microbial quality of the water is monitored regularly.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.

Disinfection and maintenance of an acceptable level of asepsis in spacecraft potable water delivery systems is a formidable task. The major area of research for this project has been to monitor the formation and growth of biofilm, and biofilm attached microorganisms, on stainless steel surfaces (specifically coupons), and the use of ozone for the elimination of these species in a closed loop system. A number of different techniques have been utilized during the course of a typical run. Scraping and sonication of coupon surfaces with subsequent plating as well as epifluorescence microscopy have been utilized to enumerate biofilm protected Pseudomonas aeruginosa. In addition, scanning electron microscopy is the method of choice to examine the integrity of the biofilm. For ozone determinations, the indigo decolorization spectrophotometric method seems most reliable. Both high- and low-nutrient cultured P. aeruginosa organisms were the target species for the ozone disinfection experiments.

Microbial proliferation in the STS potable water system is prevented by maintaining a 2-5 ppm iodine residual. The iodine is added to fuel cell water by an iodinated ion exchange resin in the Microbial Check Valve (MCV). Crew comments indicated excessive iodine in the potable water. To better define the problem, a method of in-flight iodine analysis was developed. Inflight analysis during STS-30 and STS-28 indicated iodine residuals were generally in the 9-13 ppm range. It was determined that the high iodine residual was caused by MCV influent temperatures in excess of 120 °F. This is well above the MCV operating range of 65-90 °F. The solution to this problem was to develop a resin suitable for the higher temperatures. Since 8 months were required to formulate a MCV resin suitable for the higher temperatures, a temporary solution was necessary. Two additional MCV's were installed on the chilled and ambient water lines leading into the galley to remove the excess iodine.

The purpose of this research is to determine the survival of human pathogens within a water distribution system proposed for the orbiting space station. Initially we investigated the survival of opportunistic pathogenic microorganisms in water under nutrient limiting conditions. A strain of Pseudomonas aeruginosa and two strains of Staphylococcus aureus were grown to mid-log phase then transferred to a starvation regime of sterile deionized water. Cultures were incubated at 10°, 25° or 37° C and were sampled at 24 hr, 1 week, 4 weeks and 6 weeks. The viable cell density was determined by enumerating colony forming units and by directly counting cells stained with acridine orange. Neither of the Staphylococcus strains tested were detected after 1 week of starvation. Our data indicate that Pseudomonas aeruginosa can survive in deionized water at all three temperatures tested at levels exceeding 104 cells per ml.

The Federal Government is increasingly interested in potential commercial applications for technology developed under Government funding. This emphasis has been especially evident in the Small Business Innovative Research (SBIR) Program. Experiences in commercialization efforts are presented. Benefits as well as hindrances of Government policies and regulations are discussed. Suggestions are presented based on a Small Business perspective as well as that of a Government Technical Monitor. Experience has proven that advancement to Phase III commercialization is a slow tedious process and is dependent upon a number of variables. Some technologies, such as communications, microprocessing, and medical systems are directly marketable both inside and outside of the Government.

The primary source of potable water planned for the International Space Station will be generated from the reclamation of humidity condensate, urine, and hygiene waters. It is vital to crew health and performance that this reclaimed water be safe for human consumption, and that health risks associated with recycled water consumption be identified and quantified. Only recently has data been available on the chemical constituents in reclaimed waters generated in microgravity. Results for samples collected during Mir-21 reveal that both the reclaimed water and stored water are of potable quality, although the samples did not meet U.S. standards for total organic carbon (TOC), total phenols, and turbidity.

The Microbial Check Valve (MCV) is used on the Space Shuttle to impart an iodine residual to the drinking water to maintain microbial control. Approximately twenty MCV locations have been identified in the Space Station Freedom design, each with a 90 day life. This translates to 2400 replacement units in 30 years of operation. An in situ regeneration concept has been demonstrated that will reduce this replacement requirement to less than 300 units based on data to date and potentially fewer as further regenerations are accomplished. A totally automated system will result in significant savings in crew time, resupply requirements and replacement costs. An additional feature of the device is the ability to provide a concentrated biocide source (200 mg/liter of I2) that can be used to superiodinate systems routinely or after a microbial upset. This program was accomplished under NASA Contract Number NAS9-18113.

An important milestone in the ongoing effort by NASA to develop and refine closed-loop water recycling systems for human space flight was reached during the summer of 1996 with the successful completion of Phase II of the Lunar Mars Life Support Testing Program at Johnson Space Center. Part of Phase II involved testing a water-recycling system in a closed test chamber continuously occupied by four human subjects for thirty days. The Phase II crew began the test with a supply of water that had been processed and certified for human use. As the test progressed, humidity condensate, urine, and wastewater from personal hygiene and housekeeping activities were reclaimed and reused several times. Samples were collected from various points in the reclamation process during the thirty day test. The data verified the water-processing hardware can reliably remove wastewater contaminants and produce reclaimed water that meets NASA standards for hygiene- and potable-quality water.

The Microbial Check Valve (MCV) is a reloadable flow-through canister containing iodinated ion exchange resin, which is used aboard the Shuttle Orbiter as a disinfectant to maintain water potability. The MCV exhibits a significant contact kill and imparts a biocidal residual I2 concentration to the effluent. MCVs in current use have nominal 30 day lives. MCVs baselined for Space Station Freedom will have 90 day lives, and will require replacement 120 times over 30 years. Means to extend MCV life are desirable to minimize resupply penalties. New technology has been developed for fully autonomous in situ regeneration of an expended MCV canister. The Regenerative Microbial Check Valve (RMCV) consists of an MCV, a packed bed of crystalline I2, a flow diverter valve, an in-line iodine monitor and a microcontroller. During regeneration, flow is directed first through the packed I2 bed and then into the MCV where the resin is replenished.

When construction of Space Station Freedom reaches the Permanent Manned Capability stage, plans call for the Water Recovery and Management Subsystem to treat distilled urine, spent hygiene water, and humidity condensate in order to reclaim water of potable quality. The reclamation technologies currently baselined to process these wastewaters include adsorption, ion exchange, catalytic oxidation, and disinfection. To ensure that baselined technologies will be able to effectively remove those compounds that present health risks to the crew, the National Research Council has recommended that additional information be gathered on specific contaminants in wastewaters representative of those to be encountered on Space Station. This paper reports the efforts by the Water and Food Analytical Laboratory at the Johnson Space Center to enlarge the database of potential contaminants in humidity condensate.

The Microbial Check Valve (MCV) is a canister containing an iodinated ion exchange resin and is used on the Shuttle Orbiter to provide microbial control of potable water. The MCV provides a significant contact kill, and imparts a biocidal iodine residual to the water. The Orbiter MCV has a design life of 30 days. For longer duration applications, such as Space Station Freedom, an extended life is desirable to avoid resupply penalties. A method of in situ MCV regeneration with elemental iodine is being developed. During regeneration water en route to the MCV first passes through a crystalline iodine bed where a concentration between 200 - 300 mg/L I2 is attained. When introduced into the MCV, this high concentration causes an equilibrium shift towards iodine loading, effecting regeneration of the resin. After regeneration normal flow is re-established. Life cycle regeneration testing is currently in progress.

Biofilms form rapidly on surfaces in contact with the water reclamation systems proposed for long-term space travel. These microbial populations pose a potentially serious hazard to the astronauts, the engineering systems and to the function of plant growth chambers. Human pathogens present in grey water survive for long periods on surfaces, protected against disinfection by the presence of a biofilm matrix. In addition, some of the materials being proposed for use in water reclamation systems for the orbiting space station are highly susceptible to microbial degradation. Both blockage and corrosion can occur as a result of the interactions between the microorganisms and the materials. Chemicals produced by microbial processes during growth on surfaces include both metals and organic toxins. These by-products of metabolic activity are potential hazards to astronaut health and growth of plants being considered as part of the life support system.

A significant decline in molecular iodine concentration is associated with the iodination of heavily contaminant-laden process water streams such as humidity condensate and urine distillate. Iodine loss is attributable to the reaction of this biocide with organic constituents. This phenomenon has been investigated using time resolved molecular absorption spectrophotometry of iodinated ersatz humidity condensates and iodinated ersatz urine distillates across the ultraviolet and visible spectral regions. Rates of iodine loss have also been studied using single contaminant systems at equivalent concentrations. The predominant reactive species have been identified as thiourea and formic acid. Pseudo-first order rate constants have been determined for ersatz contaminant model mixtures and for individual reactive constituents. Second order rate constants have been determined for the bimolecular reaction of iodine and formic acid.

Two simulated spacecraft water systems are being used to evaluate the effectiveness of iodine for controlling microbial contamination within such systems. An iodine concentration of about 2.0 mg/L is maintained in one system by passing ultrapure water through an iodinated ion exchange resin. Stainless steel coupons with electropolished and mechanically-polished sides are being used to monitor biofilm formation. Results after three years of operation show a single episode of significant bacterial growth in the iodinated system when the iodine level dropped to 1.9 mg/L. This growth was apparently controlled by replacing the iodinated ion exchange resin, thereby increasing the iodine level. The second batch of resin has remained effective in controlling microbial growth down to an iodine level of 1.0 mg/L. Scanning electron microscopy indicates that the iodine has impeded but may have not completely eliminated the formation of biofilm.

Controlling microbial growth and biofilm formation in spacecraft water-distribution systems is necessary to protect the health of the crew. Methods to decontaminate the water system in flight may be needed to support long-term missions. We evaluated the ability of iodine and ozone to kill attached bacteria and remove biofilms formed on stainless steel coupons. The biofilms were developed by placing the coupons in a manifold attached to the effluent line of a simulated spacecraft water-distribution system. After biofilms were established, the coupons were removed and placed in a treatment manifold in a separate water treatment system where they were exposed to the chemical treatments for various periods. Disinfection efficiency over time was measured by counting the bacteria that could be recovered from the coupons using a sonication and plate count technique. Scanning electron microscopy was also used to determine whether the treatments actually removed the biofilm.

A computer interfaced instrument designed to quantify ammonia and other nitrogenous species that are important regenerative life support water quality parameters is described in this paper. Measurements can be made either discretely by Flow Injection Analysis (FIA) or continuously using a split stream from a process flow. The monitor exhibits a predictable quadratic response for ammonia injections between 10 μg/L to 20 mg/L with a response time less than six minutes. A full description of the ammonia monitor's response to a variety of challenge solutions including real sewage treatment plant effluent is given. In addition, the response time characteristics, the effects of temperature, and interferences are described.

One of the proposed methods for monitoring the microbial quality of the water supply aboard the International Space Station is membrane filtration. We adapted this method for space flight by using an off-the-shelf filter unit developed by Millipore. This sealed unit allows liquid to be filtered through a 0.45 μm cellulose acetate filter that sits atop an absorbent pad to which growth medium is added. We combined a tetrazolium dye with R2A medium to allow microbial colonies to be seen easily, and modified the medium to remain stable over 70 weeks at 25°C. This hardware was assembled and tested in the laboratory and during parabolic flight; a modified version was then flown on STS-66. After the STS-66 mission, a back-up plastic syringe and an all-metal syringe pump were added to the kit, and the hardware was used successfully to evaluate water quality aboard the Russian Mir space station.

The ability to aseptically remove samples and products, and the capability for addition of materials to sterile or otherwise microbially susceptible systems have always been compromised by the lack of a reliable means of sterilizing the mating fixtures. Cultures of mammalian cells are particularly vulnerable to microbial contamination due to the complexity of nutrient media and the lengthy periods required for cell growth. The Microwave Sterilizable Access Port has been developed to overcome this limitation. The system consists of three primary components: a microwave power source, a combined sterilization chamber/in-line valve port assembly, and a specimen transfer interface. Microwave energy is transmitted via coaxial cable to a small pressurized chamber that serves as a sterile transition between the surrounding environment and the system during transfer of materials.

A Total Organic Carbon (TOC) Analyzer has been developed for a Life Sciences Risk Mitigation Flight Experiment to be conducted on Spacehab and the Russian space station, Mir. Initial launch is scheduled for December 1996 (flight STS-81). The analyzer will be tested on the Orbiter in the Spacehab module, including when the Orbiter is docked at the Mir space station. The analyzer is scheduled to be launched again in May 1997 (STS-84) when it will be transferred to Mir. During both flights the analyzer will measure the quality of recycled and ground-supplied potable water on the space station. Samples will be archived for later return to the ground, where they will be analyzed for comparison to in-flight results. Water test samples of known composition, brought up with the analyzer, also will be used to test its performance in microgravity. Ground-based analyses of duplicates of those test samples will be conducted concurrently with the in-flight analyses.