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Reference cultures of 16 microorganisms obtained from the American Type Culture Collection and four clinical isolates were used in standardized solutions to inoculate 60 cards for each test strain. A set of three ID and three susceptibility cards was processed in the Vitek AutoMicrobic System (AMS) immediately after inoculation. The remaining cards were refrigerated at 4°C, and sets of six cards were removed and processed periodically for up to 17 days. The preinoculated AMS cards were evaluated for microorganism identification, percent probability of correct identification, length of time required for final result, individual substrate reactions, and antibiotic minimal inhibitory concentration (MIC) values. Results indicate that 11 of the 20 microbes tested withstood refrigerated storage up to 17 days without detectable changes in delineating characteristics. MIC results appear variable, but certain antibiotics proved to be more stable than others.

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.

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.

With the restructure and funding changes for Space Station Freedom, the Environmental Health System (EHS)/Microbiology Subsystem revised its scheduling and operational requirements for component hardware. The function of the Microbiology Subsystem is to monitor the environmental quality of air, water, and internal surfaces and, in part, crew health on board Space Station. Its critical role shall be the identification of microbial contaminants in the environment that may cause system degradation, produce unsanitary or pathogenic conditions, or reduce crew and mission effectiveness. EHS/Microbiology operations and equipment shall be introduced in concert with a phased assembly sequence, from Man Tended Capability (MTC) through Permanently Manned Capability (PMC). Effective Microbiology operations and subsystem components will assure a safe, habitable, and useful spacecraft environment for life sciences research and long-term manned exploration.