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In-flight contamination control has been an important concern of NASA since the first manned missions. Previous experience has shown that uncontrolled growth of bacteria and fungi can have a detrimental effect on both the health of the crew and the proper operation of flight hardware. It is therefore imperative to develop a safe, effective method of microbial control. Spacecraft application dictates a more stringent set of requirements for biocide selection than is usually necessary for terrestrial situations. Toxicity of the biocide is the driving factor for disinfectant choice in spacecraft. This concern greatly reduces the number and types of chemical agents that can be used as disinfectants. Currently, four biocide candidates (hydrogen peroxide, quaternary ammonium compounds, iodine, glutaraldehyde) are being evaluated as potential surface disinfectants for Space Station Freedom.

Weightlessness, cosmic radiation and other space flight related conditions may adversely impact the physiology and immune status of the crew. Since microorganisms will surely be present in space habitats, the effects of space on microbial metabolic and physiologic functions will depend upon environmental conditions, types of organisms, and the duration of the flight. Because humans will conduct long-duration space missions, space microbiology must address the effect of alterations in microbial function during space flight. Even innocuous microorganisms and endogenous flora may become etiologic agents for disease during long missions. The microbial population in the closed environments of spacecraft may also become a source of toxic metabolites or the biodegradation of materials. This paper reviews studies concerning microbial behavior in closed environments, simulated microgravity, and actual space flight.

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 Six liters are withdrawn daily and the chemical and microbial quality of the water is monitored regularly.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

One of the unique aspects of the Space Station is that it will be a totally encapsulated environment and the air and water supplies will be reclaimed for reuse. The Environmental Health System, a subsystem of CHeCS (Crew Health Care System), must monitor the air and water on board the Space Station Freedom to verify that the quality is adequate for crew safety. Specifically, the Water Quality Subsystem will analyze the potable and hygiene water supplies regularly for organic, inorganic, particulate, and microbial contamination. The equipment selected to perform these analyses will be commercially available instruments which will be converted for use on board the Space Station Freedom. Therefore, the commercial hardware will be analyzed to identify the gravity dependent functions and modified to eliminate them.

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 Space Station Freedom (SSF), with a 30-year projected lifetime and a completely closed-loop Environmental Control and Life Support System (ECLSS), is perhaps the ultimate “Tight Building.” Recognizing the potential for the development of “Tight Building Syndrome” (TBS), and initiating actions to minimize possible TBS occurrences on SSF, requires a multidisciplinary approach that begins with appropriate design concerns and ends with detection and control measures on board SSF. This paper will present a brief summary of current experience with TBS on Earth. While many of the circumstances and methodologies garnered from investigating tight buildings on Earth are similar to those that might be encountered aboard SSF, the Station also presents a unique environment and a special set of constraints which will require an adaptation of previous protocols. Air contamination, including volatile organic compounds and microorganisms, will be the focus of the discussion.

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.

To satisfy a requirement to supply water to Mir station, a process for treating iodinated water on the Shuttle was developed and implemented. The treatment system consists of packed columns for removing iodine and a syringe-based injection system for adding ionic silver, the biocide used in Mir water. Technical and potable grade water is produced and transferred in batches using collapsible 44-liter contingency water containers (CWCs). Silver is added to the water via injection of a solution from preloaded syringes. Minerals are also added to water destined for drinking. During the previous four Shuttle-Mir docking missions a total of 2781 liters (735 gallons) of water produced by the Shuttle fuel cells was processed using this method and transferred to Mir. To verify the quality of the processed water, samples were collected during flight and returned for chemical analysis.

The microgravity environment of space introduces a major new variable for consideration that will affect the design and operation of bioreactors. Adequate aeration for aerobic bioreactors will be a challenge as will gas/liquid separation, removal of carbon dioxide and other bacterial metabolic waste products, control algorithms, and overall performance assessment. These challenges must be addressed in order to fully assess the efficacy of biological approaches to the recovery of potable water from wastewater in microgravity. The first step in this process is to define the fermentation parameters of the organism or consortia that will be used in these space bioreactors. This study was designed to investigate the ability of bacteria to degrade the space station detergent IGEPON.

Archival sampling of potable water and condensate for ground laboratory analysis has been an important part of the Shuttle-Mir program because of coolant leaks and other events on Mir that have affected water quality. We report here the development of and preliminary results from a novel device for single phase humidity condensate collection at system pressures. The sampler consists of a commercial-off-the-shelf Teflon® bladder and a custom reinforced Nomex® restraint that is sized properly to absorb the stress of applied pressures. A plastic Luer-Lock disconnect, with poppet actuated by a mating Luer-Lock fitting, prevents the contents from being spilled during transport. In principle, a sampler of any volume can be designed. The empty mass of the reusable one-liter sampler is only 63 grams. Several designs were pressure tested and found to withstand more than 3 atmospheres well in excess of typical spacecraft water or wastewater system pressures.

The active control of problematic microbial populations aboard spacecraft, and within future lunar and planetary habitats is a fundamental Advanced Life Support (ALS) requirement to ensure the long-term protection of crewmembers from infectious disease, and to shield materials and equipment from biofouling and biodegradation. The development of effective antimicrobial coatings and materials is an important first step towards achieving this goal and was the focus of our research. A variety of materials were coated with antibacterial and antifungal agents using covalent linkages. Substrates included both granular media and materials of construction. Granular media may be employed to reduce the number of viable microorganisms within flowing aqueous streams, to inhibit the colonization and formation of biofilms within piping, tubing and instrumentation, and to amplify the biocidal activity of low aqueous iodine concentrations.

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.

Disinfection of water and wastewater was proven to be feasible using a Breadboard Electrochemical Ozone Generator (EOG). A static gas/liquid separator, containing a microporous, hydrophobic membrane, was tested with the Breadboard EOG, and was found to increase the concentration of the ozone (O3) dissolved in the water. Distilled water and selected wastewaters were disinfected, achieving dissolved O3 concentrations up to 3 mg/L. The hardware is capable of operating in 0-g and 1-g environments. An end-item Electrochemical Ozonator (EO), sized to disinfect 116 kg of potable water per day, was projected to weigh 1.2 kg and consume only 18.5 W.

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.

Clinical-microbiological investigations are an important aspect of the crew health stabilization program. To ensure that space crews have neither active nor latent infections, clinical specimens, including throat and nasal swabs and urine samples, are collected at 10 days (L-10) and 2 days (L-2) before launch, and immediately after landing (L+0). All samples are examined for the presence of bacteria and fungi. In addition, fecal samples are collected at L-10 and examined for bacteria, fungi and parasites. This paper describes clinical-microbiological findings from 144 astronauts participating in 25 Space Shuttle missions spanning STS-26 to STS-50. The spectrum of microbiological findings from the specimens included 25 bacterial and 11 fungal species. Among the bacteria isolated most frequently were Staphylococcus aureus, Enterobacter aerogenes, Enterococcus faecalis, Escherichia coli, Proteus mirabilis and Streptococcus agalactiae.

Space flight and related factors such as stress appear to have an adverse effect on astronauts' immune systems. The presence of potentially pathogenic microbes including several genera of fungi reported from spacecraft environment may be a cause of concern in such situations. In order to study the role of such organisms in causing opportunistic or allergic diseases in crewmembers, we have tried to develop an animal model. BALB/c mice were suspended upside down for varying periods of time to induce stress, and their lymphocyte functions were evaluated. These studies indicate that the stress resulted in lowered mitogen induced lymphocyte stimulation as represented by 3H-thymidine uptake. We have also studied the ability of these animals to respond to Aspergillus fumigatus spores. The results of the study clearly demonstrate a definite down-regulation in T-cell proliferation and a higher incidence of infection with A. fumigatus.