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NASA NTRS1993

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↗

NASA NTRSDate not supplied

In-Situ Electrochemical Generation and Utilization of Hydrogen Peroxide for Disinfection

Disinfection needs to meet personal hygiene requirements at the International Space Station (ISS) is currently accomplished through the use of pre-packaged, disposable, wetted wipes, which represent an appreciable carry-along mass and disposal burden. However, as human missions travel further into the solar system the availability of resources to resupply will be diminished. Therefore, next-generation system should use onboard utilities to create on demand disinfectants, thereby reducing the dependence on earth-based supplies and further eliminating storage and disposal problems. Accordingly, we are developing an in-situ approach to electrochemically generate hydrogen peroxide disinfecting solution utilizing onboard life support supplies (Air/Water) to neutralize surface microorganisms present in closed living systems. As discussed within our 2019 and 2021 ICES papers (ICES-2019-38 and ICES-2021-273), we have continued to improve our TRL by scaling the electrochemical generation production process and validating the system in a zero-gravity parabolic loop flight test. In this paper we will demonstrate a system that can achieve over 1 L of >2 w/w% peroxide per day with DI water and air feeds. The hydrogen peroxide solutions generated were sent to NASA for microbial control property characterization. Overall, the electrochemical peroxide generation system offers a more economical and practical alternative, with the disinfectant being generated on demand and in-situ (using available life support materials of Air/Water); and applied to reusable cloths. The specific application of interest to this program is crew contact surfaces in space vehicles, but this technology could be utilized for waste water disinfection, heat exchanger biofouling remediation, and laundry applications. The peroxide generation system will also be able to address Earth-based needs in various settings such as field hospitals, restaurants, military, warehouses, movie theatres, among many others.

hydrogen peroxide↗

NASA NTRS1973

New formaldehyde base disinfectants.

Preparations of formaldehyde in various organic liquids - ethylene glycol, glycerol, and propylene glycol - serve as effective disinfectants towards microbial vegetative cells and spores. This disinfection is a temperature-dependent process and is manifest when these formaldehyde base disinfectants are dissolved in water. The irritating vapors associated with formaldehyde disinfection are not present in either of these new formaldehyde base disinfectants or in aqueous solutions of them.

Trujillo, R.↗

NASA NTRS2009

Efficacy of Various Chemical Disinfectants on Biofilms Formed in Spacecraft Potable Water System Component

As the provision of potable water is critical for successful habitation of the International Space Station (ISS), life support systems were installed in December 2008 to recycle both humidity from the atmosphere and urine to conserve available water in the vehicle. Pre-consumption testing from the dispensing needle at the Potable Water Dispenser (PWD) indicated that bacterial concentrations exceeded the current ISS specifications of 50 colony forming units (CFU) per ml. Subsequent investigations revealed that a corrugated stainless steel flex hose upstream of the dispensing needle in the PWD was filled with non-sterile water and left at room temperature for over one month before launch. To simulate biofilm formation that was suspected in the flight system, sterile flex hoses were seeded with a consortium of bacterial isolates previously recovered from other ISS water systems, which included Ralstonia pickettii, Burkholderia multivorans, Caulobacter vibrioides., and Cupriavidus pauculus. After 5 days of incubation, these hoses were challenged with various chemical disinfectants including hydrogen peroxide, colloidal silver, and buffered pH solutions to determine the ability of the disinfectants to decrease and maintain bacterial concentrations below ISS specifications. Disinfection efficacy over time was measured by collecting daily heterotrophic plate counts following exposure to the disinfectants. A single flush with either 6% hydrogen peroxide solution or a mixture of 3% hydrogen peroxide and 400 ppb colloidal silver effectively reduced the bacterial concentrations to less than 1 CFU/ml for a period of up to 2 months. Testing results indicated that hydrogen peroxide and mixtures of hydrogen peroxide and colloidal silver have tremendous potential as alternative disinfectants for ISS water systems.

Wong, Willy↗

NASA NTRS2011

Evaluation of Electrochemically Generated Potable Water Disinfectants for Use on the International Space Station

Microbial contamination and subsequent growth in spacecraft water systems are constant concerns for missions involving human crews. The current potable water disinfectant is iodine; however, with the end of the Space Shuttle program, there is a need to develop redundant biocide systems which are less dependent on hardware that would need to be launched on a regular basis. Three systems for electrochemical production of potable water disinfectants are being assessed for use on the International Space Station (ISS). Since there is a wide variability in the literature with regards to efficacy in both concentration and exposure time of these disinfectants, there is a need to establish baseline efficacy values. This paper describes a series of tests performed in order to establish optimal concentrations and exposure times for four disinfectants against single and mixed species planktonic and biofilm bacteria and to determine whether these electrochemical disinfection devices are able to produce a sufficient amount of chemical in both concentration and volume to act as a biocide for potable water on ISS.

Vega, Leticia↗

NASA NTRS1974

Corrosion control and disinfection studies in spacecraft water systems

Disinfection and corrosion control in the water systems of the Saturn 5 Orbital Workshop Program are considered. Within this framework, the problem areas of concern are classified into four general areas: disinfection; corrosion; membrane-associated problems of disinfectant uptake and diffusion; and taste and odor problems arising from membrane-disinfectant interaction.

Shea, T. G.↗

NASA NTRS2002

Effects of Disinfectants in Water on Mir- and Earth-Grown Wheat

Iodine and silver fluoride are used to purify water onboard U. S. Shuttles and the Russian Space Station, Mir, respectively. In 1995, iodine-treated water, which ranged from 1.0-4.0 mg x kg(exp -1) with a mean of 2.9 mg x kg(exp -1), was applied to Super Dwarf wheat (Triticum aestivum L.) plants when Mir water (grey or tech grade) became scarce. The potential phytotoxicity of iodine on Super Dwarf wheat is an unknown. Since use of iodine-treated water was not part of the experiment, we sought to determine whether it accounted for the subsequent poor wheat seedling growth and floral development onboard the Mir. Super Dwarf wheat seeds were imbibed in iodine or silver fluoride concentrations of 0.0, 1.0, 2.0, 4.0, 8.0 or 16.0 mg x kg(exp -1) for 96 h at 4 C. Five seeds were then planted per 13.3 cm x 13.3 cm pots containing a granular clinoptilolite (Cp) zeolite (1 -2 mm dia.) and placed in Percival(TM) growth chambers programmed for 20/15 C and 18/6 h d/n regime. Plants were irrigated with distilled water, and Iodine- or silver fluoride-treated distilled water. In separate experiments, seeds were treated as above and germination and early seedling growth were determined by examining seedling responses to disinfectants in rolled paper towels. Silver fluoride had very little effect on wheat seed germination. By contrast, iodine reduced germination at all treatment levels. Seedlings exposed to 1.0, 2.0, and 4.0 mg x kg(exp -1) of iodine or silver fluoride levels exhibited a slight stimulation in shoot and root growth. Both disinfectants at 8 and 16 mg x kg(exp -1) showed significantly (p is less than or equal to 0.01) reduced seedling shoot and root lengths and fresh biomasses compared to the control and lower disinfectant levels. The number of spikelets per spike, florets per spikelet, seeds per spike and seed weight were also significantly reduced at the 8 and 16 mg x kg(exp -1) compared to the control and lower levels of disinfectant. Based on these ground-based post-flight analyses, the levels of iodine- and/or silver fluoride-treated water used on Mir-grown plants onboard the Mir did not cause the poor growth and development of the wheat plants.

Campbell, William .F.↗

NASA NTRS2010

Pilot Study for UVA-LED Disinfection of Escherichia coli in Water for Space and Earth Applications

To test the efficacy of UVA-LED disinfection, a solution of Escherichia coli was pumped through a modified drip flow reactor at a flow rate of 1 ml/min. The experiment was conducted in a controlled environment chamber to ensure that temperature did not cause disinfection. The reactor featured three wells with different treatments: UVA-LED irradiation, UVA-LEDs with Ti02, and UVA-LEDs with nanosilver. Samples from each well were taken throughout a 340 hour period, inactivated, assayed, and analyzed for E. coli disinfection. Results of the duplicate experiments indicated longer exposure times are needed for UVA-LED disinfection of E. coli in water. Further research would consider a longer sampling period and different test conditions, such as increased contact area and various flow rates.

Ragolta, Carolina↗

NASA NTRS2011

Disinfection of Spacecraft Potable Water Systems by Photocatalytic Oxidation Using UV-A Light Emitting Diodes

Ultraviolet (UV) light has long been used in terrestrial water treatment systems for photodisinfection and the removal of organic compounds by several processes including photoadsorption, photolysis, and photocatalytic oxidation/reduction. Despite its effectiveness for water treatment, UV has not been explored for spacecraft applications because of concerns about the safety and reliability of mercury-containing UV lamps. However, recent advances in ultraviolet light emitting diodes (UV LEDs) have enabled the utilization of nanomaterials that possess the appropriate optical properties for the manufacture of LEDs capable of producing monochromatic light at germicidal wavelengths. This report describes the testing of a commercial-off-the-shelf, high power Nichia UV-A LED (250mW A365nnJ for the excitation of titanium dioxide as a point-of-use (POD) disinfection device in a potable water system. The combination of an immobilized, high surface area photocatalyst with a UV-A LED is promising for potable water system disinfection since toxic chemicals and resupply requirements are reduced. No additional consumables like chemical biocides, absorption columns, or filters are required to disinfect and/or remove potentially toxic disinfectants from the potable water prior to use. Experiments were conducted in a static test stand consisting of a polypropylene microtiter plate containing 3mm glass balls coated with titanium dioxide. Wells filled with water were exposed to ultraviolet light from an actively-cooled UV-A LED positioned above each well and inoculated with six individual challenge microorganisms recovered from the International Space Station (ISS): Burkholderia cepacia, Cupriavidus metallidurans, Methylobacterium fujisawaense, Pseudomonas aeruginosa, Sphingomonas paucimobilis and Wautersia basilensis. Exposure to the Nichia UV-A LED with photocatalytic oxidation resulted in a complete (>7-log) reduction of each challenge bacteria population in <180 minutes of contact time. With continued advances in the design and manufacture of UV-A LEDs and semi-conducting photocatalysts, LED activated photochemical process technology promises to extend its application to spacecraft environmental systems.

Birmele, Michele N.↗

NASA NTRS2012

Evaluation of Electrochemically Generated Potable Water Disinfectants for Use on the International Space Station

Microbial contamination and subsequent growth in spacecraft water systems are constant concerns for missions involving human crews. The current potable water disinfectant for the International Space Station (ISS) is iodine; however, with the end of the Space Shuttle program, there is a need to develop redundant biocide systems that do not require regular up ]mass dependencies. Throughout the course of a year, four different electrochemical systems were investigated as a possible biocide for potable water on the ISS. Research has indicated that there is a wide variability with regards to efficacy in both concentration and exposure time of these disinfectants, therefore baseline efficacy values were established. This paper describes a series of tests performed in order to establish optimal concentrations and exposure times for four disinfectants against single and mixed species planktonic and biofilm bacteria. Results of the testing determined whether these electrochemical disinfection systems are able to produce a sufficient amount of chemical in both concentration and volume to act as a biocide for potable water on ISS.

Rodriquez, Branelle↗

NASA NTRS2012

Material Testing in Support of the ISS Electrochemical Disinfection Feasibility Study

Rodriquez, Branelle↗

NASA NTRS2013

Evaluation of Electrochemically Generated Potable Water Disinfectants for Use on the International Space Station

Microbial contamination and subsequent growth in spacecraft water systems are constant concerns for missions involving human crews. The current potable water disinfectant for the International Space Station (ISS) is iodine; however, with the end of the Space Shuttle Program, there is a need to develop redundant biocide systems that do not require regular up-mass dependencies. Throughout the course of a year, four different electrochemical systems were investigated as a possible biocide for potable water on the ISS. Research has indicated that a wide variability exists with regards to efficacy in both concentration and exposure time of these disinfectants; therefore, baseline efficacy values were established. This paper describes a series of tests performed to establish optimal concentrations and exposure times for four disinfectants against single and mixed species planktonic and biofilm bacteria. Results of the testing determined whether these electrochemical disinfection systems are able to produce a sufficient amount of chemical in both concentration and volume to act as a biocide for potable water on the ISS.

Rodriquez, Branelle↗

NASA NTRSDate not supplied

Literature Review of Disinfection Techniques For Water Treatment

Water treatment is a developing concern, both terrestrially and in spacecraft, as exploration missions extend in time and distance. Current biofilm control is limited for long-term applications. To optimize biocides for present and future space exploration vehicles, a thorough understanding of common and traditional disinfectant techniques is required. This review is focused on the three fundamental disinfection techniques: chemical, physical, and biological. Mechanisms, advantages, disadvantages, and specific properties of each major technique, as well as their studied effect on established biofilms, are also considered. This paper provides a general background on disinfectants and some information on effects on biofilms that can be useful to develop innovative ideas for state-of-the-art disinfection techniques for water treatment in specific environments, such as those currently posing mission risks as well as for future spacecraft water system development.

Nilab Azim↗

NASA NTRS1987

Review of water disinfection techniques

Throughout the history of manned space flight the supply of potable water to the astronauts has presented unique problems. Of particular concern has been the microbiological quality of the potable water. This has required the development of both preflight water system servicing procedures to disinfect the systems and inflight disinfectant addition and monitoring devices to ensure continuing microbiological control. The disinfectants successfully used to date have been aqueous chlorine or iodine. Because of special system limitations the use of iodine has been the most successful for inflight use and promises to be the agent most likely to be used in the future. Future spacecraft potable, hygiene, and experiment water systems will utilize recycled water. This will present special problems for water quality control. NASA is currently conducting research and development to solve these problems.

Colombo, Gerald V.↗

NASA NTRS1989

Effect of iodine disinfection products on higher plants

Iodine is used to disinfect potable water on United States spacecraft. Iodinated potable water will likely be used to grow plants in space. Little is known about the effects of iodine disinfection products on plants. Seeds of select higher plants were germinated in water iodinated using the Shuttle Microbial Check Valve, and water to which measured amounts of iodine was added. Percent germination was decreased in seeds of most species germinated in iodinated water. Beans were most affected. Germination rates, determined from germination half-times, were decreased for beans germinated in iodinated water, and water to which iodide was added. Development was retarded and rootlets were conspicuously absent in bean and several other plant species germinated in iodinated water. Iodide alone did not elicit these responses. Clearly iodine disinfection products can affect higher plants. These effects must be carefully considered for plant experimentation and cultivation in space, and in design and testing of closed environmental life support systems.

Janik, D.↗

NASA NTRS1990

Application of the pentaiodide strong base resin disinfectant to the U.S. space program

A pentaiodide resin is described which has 70 percent of its weight composed of elemental iodine, has a relatively low iodine residual, and may offer superior disinfection capability for applications on long-duration space vehicles. Such a disinfectant is crucial for use on spacecraft for long periods of time where water would be recycled through various systems. The pentaiodide resin is capable of devitalizing 1 x 10 to the 9th bacteria per ml in aqueous suspension within 10 seconds of contact with the resin bed. A number of organisms have already been tested and the resin continues to prove effective. Resin properties and composition are discussed and a detailed account of the first investigation of the pentaiodide resin as a disinfectant against the intestinal parasite Giardia lamblia is provided.

Marchin, George L.↗

NASA NTRS1993

Effect of iodine disinfectant source and water quality parameters on soluble iodine speciation

Investigations of iodine species distribution of various aqueous solutions of iodine disinfectants and water from equilibrated suspensions of triodide and pentaiodide resins were done at the University of Colorado for the Center for Space Environmental Health during 1992 and 1993. Direct measurements of three individual iodine species: I(-), I2 and I3(-), were made. In addition three measures of total titratable iodine species were used. It has been found that I2 and I3(-) solutions produce a significant fraction of the non-disinfecting species iodine I(-), ranging from 50 to 80% of added iodine, respectively, at pH values of approximately 5. Correspondingly, I2 solutions produce more than twice the concentration of disinfecting iodine species per mass iodine dose than I3(-) solutions. Both I(-) and I2 species were found in aqeuous extracts of pentaiodide resin, although no soluble species were detected with triiodide resin.

Silverstein, Joann↗

NASA NTRS1999

The use of multiple indices of physiological activity to access viability in chlorine disinfected Escherichia coli O157:H7

A suite of fluorescent intracellular stains and probes was used, in conjunction with viable plate counts, to assess the effect of chlorine disinfection on membrane potential (rhodamine 123; Rh123 and bis-(1,3-dibutylbarbituric acid) trimethine oxonol; DiBAC4(3)), membrane integrity (LIVE/DEAD BacLight kit), respiratory activity (5-cyano-2,3-ditolyl tetrazolium chloride; CTC) and substrate responsiveness (direct viable counts; DVC) in the commensal pathogen Escherichia coli O157:H7. After a 5 min exposure to the disinfectant, physiological indices were affected in the following order: viable plate counts > substrate responsiveness > membrane potential > respiratory activity > membrane integrity. In situ assessment of physiological activity by examining multiple targets, as demonstrated in this study, permits a more comprehensive determination of the site and extent of injury in bacterial cells following sublethal disinfection with chlorine. This approach to assessing altered bacterial physiology has application in various fields where detection of stressed bacteria is of interest.

NASA Discipline Environmental Health↗