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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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Removal of Burkholderia cepacia biofilms with oxidants

Iodine is used to disinfect the water system aboard US space shuttles and is the anticipated biocide for the international space station. Water quality on spacecraft must be maintained at the highest possible levels for the safety of the crew. Furthermore, the treatment process used to maintain the quality of water on research must be robust and operate for long periods with minimal crew intervention. Biofilms are recalcitrant and pose a major threat with regard to chronic contamination of spacecraft water systems. We measured the effectiveness of oxidizing biocides on the removal and regrowth of Burkholderia (Pseudomonas) cepacia biofilms. B. cepacia, isolated from the water distribution system of the space shuttle Discovery, was grown in continuous culture to produce a bacterial contamination source for biofilm formation and removal studies. A 10(7) CFU ml-1 B. cepacia suspension, in distilled water, was used to form biofilms on 3000 micrometers2 glass surfaces. Rates of attachment were measured directly with image analysis and were found to be 7.8, 15.2, and 22.8 attachment events h-1 for flow rates of 20.7, 15.2, and 9.8 ml min-1, respectively. After 18 h of formation, the B. cepacia biofilms were challenged with oxidants (ozone, chlorine, and iodine) and the rates of biofilm removal determined by image analysis. Fifty percent of the biofilm material was removed in the first hour of continous treatment with 24 mg l-1 chlorine or 2 mg l-1 ozone. Iodine (48 mg l-1) did not remove any measurable cellular material after 6 h continuous contact. After this first removal of biofilms by the oxidants, the surface was allowed to refoul and was again treated with the biocide. Iodine was the only compound that was unable to remove cellular debris from either primary or secondary biofilms. Moreover, treating primary biofilms with iodine increased the rate of formation of secondary biofilms, from 4.4 to 5.8 attachment events h-1. All the oxidants tested inactivated the B. cepacia associated with both primary and secondary biofilms. The amount of biocide needed to inactivate 50% of planktonic B. cepacia in 10 min at 25 degrees C was 8.4, 0.5, and 0.2 mg l-1 for iodine, chlorine, and ozone, respectively. The data suggest that iodine maynot be the best chemical for treating of biofilms when removal of cellular material is required.

NASA Discipline Environmental Health↗

Ground-based studies with Super-Dwarf wheat in preparation for space flight

Several experiments were carried out to test responses of a Super-Dwarf cultivar of wheat (Triticum aestivum L.) to various environmental parameters that were anticipated to be present in our attempts to grow the wheat in a small growth chamber on the Russian Space Station, Mir, or that proved to be present in a 1995 trial space experiment. Under low photosynthetic photon flux (40-400 micromoles m-2 s-1 PPF), development (e.g. anthesis) was retarded, but heads (often sterile) always formed, even if light was so low that plants died before the heads could mature. Longer photoperiods promoted flowering, but night interruptions combined with short days did not provoke a long-day response as occurs with true long-day plants. The long-day effect could prove to be a summation of photosynthetic products. Heat stress (40 degrees C for 1-24 h) did not influence flowering but killed plants that were 13-16-day-old (no effect on younger plants). Concentrations of iodine or silver-fluoride disinfectants present in the water used for plants on Mir (1.0-4.0 mg L-1) did not affect plant growth although higher concentrations (8.0-1.6 mg L-1) were inhibitory. GA3 or indoleacetic acid applied every other day at concentrations from 1.0 x 10(-6) mg L-1 to 3.162 x 10(-4) mg L-1 did not change the height of Super-Dwarf wheat, suggesting that this cultivar is not a gibberellin mutant.

Flight Experiment↗

Health significance and occurrence of injured bacteria in drinking water

Enteropathogenic and indicator bacteria become injured in drinking water with exposure to sublethal levels of various biological, chemical and physical factors. One manifestation of this injury is the inability to grow and form colonies on selective media containing surfactants. The resulting underestimation of indicator bacteria can lead to a false estimation of water potability. m-T7 medium was developed specifically for the recovery of injured coliforms (both "total" and fecal) in drinking water. The m-T7 method was used to survey operating drinking water treatment and distribution systems for the presence of injured coliforms that were undetected with currently used media. The mean recovery with m-Endo LES medium was less than 1/100 ml while it ranged between 6 and 68/100ml with m-T7 agar. The majority of samples giving positive results with m-T7 medium yielded no detectable coliforms with m-Endo LES agar. Over 95% of the coliform bacteria in these samples were injured. Laboratory experiments were also done to ascribe the virulence of injured waterborne pathogens. Enteropathogens including Salmonella typhimurium, Yersinia enterocolitica and Shigella spp. required up to 20 times the chlorine levels to produce the same injury in enterotoxigenic Escherichia coli (ETEC) and nonpathogenic coliforms. Similar results were seen with Y. enterocolitica exposed to copper. The recovery of ETEC was followed by delayed enterotoxin production, both in vitro and in the gut of experimental animals. This indicates that injured waterborne enteropathogenic bacteria can be virulent.

NASA Discipline Number 13-40↗

Iodine susceptibility of pseudomonads grown attached to stainless steel surfaces

Pseudomonads were adapted to grow in phosphate-buffered water and on stainless steel surfaces to study the iodine sensitivity of attached and planktonic cells. Cultures adapted to low nutrient growth were incubated at room temperature in a circulating reactor system with stainless steel coupons to allow biofilm formation on the metal surfaces. In some experiments, the reactor was partially emptied and refilled with buffer at each sampling time to simulate a "fill-and-draw" water system. Biofilms of attached bacteria, resuspended biofilm bacteria, and reactor suspension, were exposed to 1 mg l-1 iodine for 2 min. Attached bacterial populations which established on coupons within 3 to 5 days displayed a significant increase in resistance to iodine. Increased resistance was also observed for resuspended cells from the biofilm and planktonic bacteria in the system suspension. Generally, intact biofilms and resuspended biofilm cells were most resistant, followed by planktonic bacteria and phosphate buffer cultures. Thus, biofilm formation on stainless steel surfaces within water systems can result in significantly increased disinfection resistance of commonly-occurring water-borne bacteria that may enhance their ability to colonise water treatment and distribution systems.

NASA Program Environmental Health↗

A direct viable count method for the enumeration of attached bacteria and assessment of biofilm disinfection

This report describes the adaptation of an in situ direct viable count (in situ DVC) method in biofilm disinfection studies. The results obtained with this technique were compared to two other enumeration methods, the plate count (PC) and conventional direct viable count (c-DVC). An environmental isolate (Klebsiella pneumoniae Kp1) was used to form biofilms on stainless steel coupons in a stirred batch reactor. The in situ DVC method was applied to directly assess the viability of bacteria in biofilms without disturbing the integrity of the interfacial community. As additional advantages, the results were observed after 4 h instead of the 24 h incubation time required for colony formation and total cell numbers that remained on the substratum were enumerated. Chlorine and monochloramine were used to determine the susceptibilities of attached and planktonic bacteria to disinfection treatment using this novel analytical approach. The planktonic cells in the reactor showed no significant change in susceptibility to disinfectants during the period of biofilm formation. In addition, the attached cells did not reveal any more resistance to disinfection than planktonic cells. The disinfection studies of young biofilms indicated that 0.25 mg/l free chlorine (at pH 7.2) and 1 mg/l monochloramine (at pH 9.0) have comparable disinfection efficiencies at 25 degrees C. Although being a weaker disinfectant, monochloramine was more effective in removing attached bacteria from the substratum than free chlorine. The in situ DVC method always showed at least one log higher viable cell densities than the PC method, suggesting that the in situ DVC method is more efficient in the enumeration of biofilm bacteria. The results also indicated that the in situ DVC method can provide more accurate information regarding the cell numbers and viability of bacteria within biofilms following disinfection.

NASA Discipline Number 04-10↗

Effects of starvation on physiological activity and chlorine disinfection resistance in Escherichia coli O157:H7

Escherichia coli O157:H7 can persist for days to weeks in microcosms simulating natural conditions. In this study, we used a suite of fluorescent, in situ stains and probes to assess the influence of starvation on physiological activity based on membrane potential (rhodamine 123 assay), membrane integrity (LIVE/DEAD BacLight kit), respiratory activity (5-cyano-2,3-di-4-tolyl-tetrazolium chloride assay), intracellular esterase activity (ScanRDI assay), and 16S rRNA content. Growth-dependent assays were also used to assess substrate responsiveness (direct viable count [DVC] assay), ATP activity (MicroStar assay), and culturability (R2A agar assay). In addition, resistance to chlorine disinfection was assessed. After 14 days of starvation, the DVC values decreased, while the values in all other assays remained relatively constant and equivalent to each other. Chlorine resistance progressively increased through the starvation period. After 29 days of starvation, there was no significant difference in chlorine resistance between control cultures that had not been exposed to the disinfectant and cultures that had been exposed. This study demonstrates that E. coli O157:H7 adapts to starvation conditions by developing a chlorine resistance phenotype.

NASA Discipline Environmental Health↗

Effects of culture conditions and biofilm formation on the iodine susceptibility of Legionella pneumophila

The susceptibility of Legionella pneumophila to iodination was studied with cultures grown in well water, on rich agar media, and attached to stainless-steel surfaces. Legionella pneumophila grown in water cultures in association with other microorganisms were less sensitive to disinfection by chlorine and iodine than were agar-passaged cultures. Differences in sensitivity to disinfection between water-cultured and agar-grown legionellae were determined by comparing C x T values (concentration in milligrams per litre multiplied by time in minutes to achieve 99% decrease in viability) and CM x T values (concentration in molarity). Iodine (1500x) gave a greater difference in CM x T values than did chlorine (68x). Iodine was 50 times more effective than chlorine when used with agar-grown cultures but was only twice as effective when tested against water-grown Legionella cultures. C x T x S values (C x T multiplied by percent survivors), which take into consideration the percent surviving bacteria, were used to compare sensitivities in very resistant populations, such as those in biofilms. Water cultures of legionellae associated with stainless-steel surfaces were 135 times more resistant to iodination than were unattached legionellae, and they were 210,000 times more resistant than were agar-grown cultures. These results indicate that the conditions under which legionellae are grown can dramatically affect their susceptibility to some disinfectants and must be considered when evaluating the efficacy of a disinfecting agent.

Non-NASA Center↗