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Technical Paper

Evaluation of Methods for Remediating Biofilms in Spacecraft Potable Water Systems

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.
Technical Paper

Biofilm Formation and Control in a Simulated Spacecraft Water System: Three Year Results

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.
Technical Paper

Viral Populations within the International Space Station's Internal Active Thermal Control System Ground Support and Potential Flight Hardware

The Internal Active Thermal Control System (IATCS) aboard the International Space Station (ISS) contains an aqueous, alkaline fluid (pH 9.5±0.5) that aids in maintaining a habitable environment for the crew. Because microbes have significant potential to cause disease, adverse effects on astronaut health, and microbe-induced corrosion, the presence of both bacteria and viruses within IATCS fluids is of concern. This study sought to detect and identify viral populations in IATCS samples obtained from the Kennedy Space Center as a first step towards characterizing and understanding potential risks associated with them. Samples were concentrated and viral nucleic acids (NA) extracted providing solutions containing 8.87-22.67 μg NA per mL of heat transfer fluid. After further amplification viral DNA and cDNA were then pooled, fluorescently labeled, and hybridized onto a Combimatrix panvira 12K microarray containing probes for ∼1,000 known human viruses.
Technical Paper

Microbial Burden of Commercial Aircraft Cabin Air

The microbial burdens of 69 cabin air samples collected in-flight aboard commercial airliners were assessed via culture-dependent and molecular-based microbial enumeration assays. Cabin air samples from each of four separate flights aboard two different carriers were collected via air-impingement. Microbial enumeration techniques targeting DNA, ATP, and endotoxin were employed to estimate total microbial burden. The total viable microbial population ranged from 0 to 3.6 × 104 cells per 100 liters of air, as assessed by the ATP-assay. When these same samples were plated on minimal medium, anywhere from 2 to 80% of the viable population was cultivable. Five of the 29 samples examined exhibited higher cultivable plate counts than ATP-derived viable counts, perhaps a consequence of the dormant nature (lower concentration of intracellular ATP) of cells inhabiting these air cabin samples.
Technical Paper

Assessment of Microbial Community Variability in Replicate Tubular Nitrifying Bioreactors using PCR and TRFLP Analysis

Bioregenerative life support systems (BLSS) may be necessary for long-term space missions due to the high costs of lifting supplies and equipment into orbit. Much of the recycling to be done in a BLSS involves microbial activity. Although most studies to date have used a culture-based approach to characterize bacteria in BLSS under development, recently work has begun utilizing non-culture-based, DNA approaches to elucidate which microbes are present. In this study, we investigated whether replicate reactors develop replicate microbial communities using a 16S rRNA gene approach and terminal restriction length polymorphism analysis for tubular, nitrifier reactors in use at JSC. Our result suggests that both individual reactor and temporal signals can be detected in the microbial populations. This information may lead to optimization of inoculation procedures and reactor operations conditions to increase predictability and reliability of biological systems.
Technical Paper

Evaluation of Sample Preservation Methods for Space Missions

This study of samples collected from Mars 01 Orbiter was conducted to gain a better understanding and practical experience in methods to process and preserve samples intended for planetary protection analysis. Samples were evaluated for the viable growth of microbes, the molecular biomarker adenosine triphosphate (ATP), and the presence of lipopolysaccharide, a bacterial cell wall component. Losses were observed in the number of viable microbes after freezing as well as in the detectable lipopolysaccharide. Two independent studies of pooled cleanroom samples demonstrate good ATP recovery and consistent values after freezing at −20 °C.
Technical Paper

Q-PCR Based Bioburden Assessment of Drinking Water Throughout Treatment and Delivery to the International Space Station

Previous studies indicated evidence of opportunistic pathogens in samples obtained during missions to the International Space Station (ISS). This study utilized TaqMan quantitative PCR to determine specific gene abundance in potable and non-potable ISS waters. Probe and primer sets specific to the small subunit rRNA genes were designed and used to elucidate overall bacterial rRNA gene numbers. In addition, primer-probe sets specific for Burkholderia cepacia and Stenotrophomonas maltophilia were optimized and genes of these two opportunistic pathogens quantified in the pre- and post-flight drinking water as well as coolant waters. This Q-PCR approach supports findings of previous culture-based studies however; the culture based studies may have underestimated the microbial burden of ISS drinking water.
Journal Article

ATCC 29669 Spores Show Substantial Dry Heat Survivability

Bacillus sp. ATCC 29669 was isolated from microbial fallout in clean rooms during the assembly of the Viking Spacecraft missions to Mars, making it a potential contamination concern for outbound space missions. Spores from this bacterial strain were found to be thirty times more resistant to dry heat than B. atrophaeus. Spore inactivation rates under vacuum controlled humidity were faster than rates obtained under ambient humidity. Inactivation rates for these heat resistant spores are important considerations for planetary protection implementation where temperature, time and humidity conditions are used to estimate the effectiveness of dry heat microbial reduction (DHMR) procedures.