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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.

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.

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 Six liters are withdrawn daily and the chemical and microbial quality of the water is monitored regularly.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