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Solubilization studies were carried out on various cured silicone resins. A solvent spectrum was prepared. It was found that complete dissolution of cured silicone resins could be achieved without extensive physical degradation of samples. Based on the solubilization results, amine solvents were selected for spore viability studies.
Review of the knowledge available on the extent to which microorganisms (mainly microbial spores, vegetative cells, and fungi) are capable of surviving the environment of deep space, based on recent simulation experiments of deep space. A description of the experimental procedures used is followed by a discussion of deep space ecology, the behavior of microorganisms in ultrahigh vacuum, and factors influencing microbial survival. It is concluded that, so far, simulation experiments have proved far less lethal to microorganisms than to other forms of life. There are, however, wide gaps in the knowledge available, and no accurate predictions can as yet be made on the degree of lethality that might be incurred by a microbial population on a given mission. Therefore, sterilization of spacecraft surfaces is deemed necessary if induced panspermia (i.e., interplanetary life propagation) is to be avoided.
The sterilization parameters for the Viking lander of D sub 125 C = 30 minutes and z = 21 C for the exposed bioburden were derived from the experimental findings of several laboratories conducting thermal inactivation studies on Bacillus subtilis var. niger by dry heat. The moisture constraint, that the sterilizing gas shall be less than 25 percent relative humidity at standard conditions of 0 C and 760 mm Hg pressure, was added in recognition of the profound influence of water vapor on the time and temperature required for thermal inactivation of these spores. Data is presented demonstrating that the application of the moisture parameter does not significantly change the D sub 125 C and z values of 30 minutes and 21 C, respectively. Data are presented also to show the maximum influence that could be expected by decreasing the humidity to near zero percent relative humidity at 105, 113, and 125 C.
The project to evaluate thermal sterilization for unmanned landers is reported. A temperature controlled oven with a nitrogen gas supply containing a known concentration of water is discussed. The studies show that bacillus lentus, bacillus brevis, bacillus coagulans, atypical bacillus spp., and actinomycete are isolated heat survivors. The thermal resistance is given for naturally occurring airborne bacterial spores collected on exposed teflon ribbons.
Spacecraft sterilization is a vital factor in projects for the successful biological exploration of other planets. The microorganisms of major concern are the fungi and bacteria. Sterilization procedures are oriented toward the destruction of bacterial spores. Gaseous sterilants are examined, giving attention to formaldehyde, beta-propiolactone, ethylene oxide, and the chemistry of the bactericidal action of sterilants. Radiation has been seriously considered as another method for spacecraft sterilization. Dry heat sterilization is discussed together with the effects of ethylene oxide decontamination and dry heat sterilization on materials.
A determination was made of the kinds and numbers of bacteria surviving when two soils were maintained in the laboratory under dry conditions for more than half a year. Certain non-spore-forming bacteria were found to survive in the dry condition for long periods. A higher percentage of drought-tolerant than drought-sensitive bacteria was able to grow at low water activities. When they were grown in media with high salt concentrations, bacteria generally became more tolerant of prolonged drought and they persisted longer. The percent of cells in a bacterial population that remained viable when exposed to drought stress varied with the stage of growth.
A study was initiated to determine whether viable atmospheric particles such as plant pollens and fungal spores containing unsaturated lipids can interact with singlet oxygen to give oxygenated products that are potentially toxic. The results obtained confirm that surface and near surface components of common viable particulate matter in the atmosphere may be subject to rapid oxidation by singlet oxygen, leading to products which are probably allylic hydroperoxides. In connection with increasing atmospheric pollution, it is important to note that materials toxic to mammalian lung tissue may be oxidatively produced on the surfaces of viable particulate matter.
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.
Spore recovery form cured silicone potting compounds using amine solvents to degrade the cured polymers was investigated. A complete list of solvents and a description of the effect of each on two different silicone polymers is provided.
Microorganisms capable of growth at 7 C were enumerated and isolated from soil samples from the manufacture area (Denver, Colorado) and assembly area (Cape Kennedy, Florida) of the Viking spacecraft. Temperature requirements were determined for these isolates, and those growing at 3 C, but not at 32 C were designated as obligate psychrophiles in this investigation. These were identified to major generic groups, and the population density of obligate psychrophiles from the various groups was determined. Dry heat D-values were found for those spores that demonstrated growth or survival under a simulated Martian environment.
An investigation was conducted to demonstrate whether airborne microbes could propagate. The procedure consisted of: (1) looking for dilution of a labelled base in DNA; (2) looking for labelling of DNA by mixing aerosols of the label and the cells; (3) examining changes in cell size; (4) testing the possibility of spore germination; and (5) seeking evidence of an increase in cell number. Results indicate that growth and propagation can occur under special conditions, principally at temperatures of approximately 30 C (87 F) and water activity equivalents of 0.95 to 0.98.
Efforts to show growth activity of anaerobic bacteria in aerosolized particles suspended in anaerobic gas (N2) are described. Evidence of spore generation in nitrogen was obtained. Results are discussed.
Statistical techniques which have relevance to studies on the thermal inactivation on bacterial spores are discussed.
Two joint inflight biomedical experiments were conducted during the unique Apollo-Soyuz Test Project (ASTP) spaceflight. One experiment evaluated rhythmicity of spore production of Streptomyces levoris. The other evaluated components of the infectious disease process by measuring alteration in: (1) the composition of the microbial population inhabiting USA and USSR crewmembers and spacecraft; (2) the ability of each crewmember's defense mechanism to resist infection; and (3) the ability of certain microorganisms to originate infections. These two experiments are described and the major results discussed.
Results are presented for an experimental study designed to assess the effect of hydrogen peroxide on both sporeforming and nonsporeforming spacecraft isolates as an initial step in determining its suitability for microbiological decontamination of certain United States spacecraft. Survivor data were obtained for eight bacterial isolates (six sporeformers and two nonsporeformers) recovered before launch Mariner 9 and exposed to concentrations of 3, 10, and 15% hydrogen peroxide. The effects of various concentrations of hydrogen peroxide on the spores are presented in tabular form, along with the percentage of survival of nonsporeformers exposed to hydrogen peroxide. No viable vegetative cells were recovered after a 10-min exposure time to any of the three concentration of hydrogen peroxide.
Past space microbiology studies have evaluated three general areas: microbe detection in extraterrestrial materials; monitoring of autoflora and medically important species on crewmembers, equipment, and cabin air; and in vitro evaluations of isolated terrestrial species carried on manned and unmanned spaceflights. These areas are briefly reviewed to establish a basis for presenting probable experiment subjects applicable to the Space Shuttle era. Most extraterrestrial life detection studies involve visitations to other heavenly bodies. Although this is not applicable to the first series of Shuttle flights, attempts to capture meteors and spores in space could be important. Human pathogen and autoflora monitoring will become more important with increased variety among crewmembers. Inclusion of contaminated animal and plant specimens in the space lab will necessitate inflight evaluation of cross-contamination and infection potentials. The majority of Shuttle microbiology studies will doubtless fall into the third study area. Presence of a space lab will permit a whole range of experimentation under conditions similar to these experienced in earth-based laboratories. The recommendations of various study groups are analyzed, and probable inflight microbiological experiment areas are identified for the Life Sciences Shuttle Laboratory.
Planetary quarantine requirements associated with the launch of two Viking spacecraft necessitated microbiological assessment during assembly and testing at Cape Canaveral and the Kennedy Space Center. Samples were collected from the Viking Lander Capsules (VLC), Orbiters (VO), and Shrouds at predetermined intervals during assembly and testing. Levels of bacterial spores per square meter on the VLC-1 and VLC-2 were 1.6 x 10 squared and 9.7, respectively, prior to dry-heat sterilization. The ranges of aerobic mesophilic microorganisms detected on the VO-1 and VO-2 were 4.2 x 10 squared to 4.3 x 10 cubed and 2.3 x 10 squared to 8.9 x 10 cubed/sq m, respectively. Approximately 1300 colonies were picked from culture plates, identified, lyophilized, and stored for future reference. About 75% of all isolates were microorganisms considered indigenous to humans; the remaining isolates were associated with soil and dust. The percentage of microorganisms of human origin was consistent with results obtained with previous automated spacecraft but slightly lower than those observed for manned (Apollo) spacecraft.
A series of experiments at several levels of relative humidity and radiation dose rates was carried out using spores of Bacillus subtilis var. niger to evaluate the effect of heat alone, radiation alone, and a combination of heat and radiation. Combined heat and radiation treatment of microorganisms yields a destruction rate greater than the additive rates of the independent agents. The synergistic mechanism shows a proportional dependency on radiation dose rate, an Arrhenius dependence on temperature, and a dependency on relative humidity. Maximum synergism occurs under conditions where heat and radiation individually destroy microorganisms at approximately equal rates. Larger synergistic advantage is possible at low relative humidities rather than at high relative humidities.