An investigation of a sono-chemical approach in sterilization problems Final semiannual progress report no. 6, 1 Jan. - 30 Jun. 1967
Ultrasound used in combination with propylene oxide for sterilization of bacterial spores
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Ultrasound used in combination with propylene oxide for sterilization of bacterial spores
Dry heat resistance of bacterial spores from sterilization and assembly laboratory
Dry heat inactivation of bacterial spores and influence of spore moisture content on z values as related to spacecraft sanitation
Dry heat resistances of bacterial spores, and levels of microbial contamination on Apollo spacecraft
Heat shock effects on bacterial spore recovery and Apollo 9, 10, and 11 microbial contamination data
Survival of bacterial spores under various temperature and humidity conditions related to planetary quarantine
Techniques for planetary quarantine and bacterial spore inactivation
Characteristics of bacterial spore population from Cape Kennedy soil
Design, development, and applications of quartz crystal microbalance are discussed. Two types of crystals are used. One serves as reference and other senses changes in mass. Specific application to study of bacterial spores is described.
Tests were conducted to determine the dry heat resistance at 125 C of a naturally occurring bacterial spore population in a mixture of sieved vacuum cleaner dusts from Cape Kennedy. The dust was aerosolized in a special chamber and was allowed to settle on 32 Teflon ribbons to provide approximately 500,000 spores per ribbon.
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.
The mechanism for thermal inactivation of bacterial spores under moist or dry heat was studied. Experimental conditions were established relating to spore loss of heat resistance and loss of optical density as a measure of the rate and extent of germination in spore suspensions. Events occurring during germination were correlated with phase darkening (refractility and non-refractility of spores), stainability characteristics of heat and non-heat treated spores, morphological characteristics, and studies on swelling of spores by an increase in packed cell volume.
The mechanistic basis of the synergetic effect of combined heat and radiation on microbial destruction was analyzed and results show that radiation intensity, temperature, and relative humidity are the determining factors. Dry heat resistance evaluation for selected bacterial spore crops indicates that different strains of Bacillus stearothermophilus demonstrate marked differences in resistance. Preliminary work to determine the effects of storage time, suspending medium, storage temperature and spore crop cleaning procedures on dry heat survival characteristics of Bacillus subtilis var. Niger, and dry heat resistance of natural microflora in soil particles is also reported.
Statistical techniques which have relevance to studies on the thermal inactivation on bacterial spores are discussed.
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.
The radiation hazard inside spacecraft is discussed with emphasis on its effects on the crew, biological specimens, and spacecraft instruments. The problem of light flash sensations in the eyes of astronauts is addressed and experiment MA-106 is described. In this experiment, light flashes seen by blindfolded astronauts were counted and high energy cosmic ray intensity in the command module cabin were measured. The damage caused by cosmic ray hits on small living organisms was investigated in the Biostack 3 experiment (MA-107). Individual cosmic rays were tracked through layers of bacterial spores, small seeds, and eggs interleaved with layers of AgCl-crystal wafers, special plastic, and special photographic film that registered each cosmic ray particle passed.
The main objectives of the Biostack Experiment are to study the effectiveness of the structured components of the cosmic radiation to bacterial spores, plant seeds, and animal cysts for a long duration spaceflight and to get dosimetric data such as particle fluences and spectra and total doses for the Long Duration Exposure Facility orbit. The configuration of the experiment packages allows the localization of the trajectory of the particles in each biological layer and to correlate the potential biological impairment or injury with the physical characteristics of the responsible particle. Although the Biostack Experiment was designed for a long duration flight of only nine months, most of the biological systems show a high hatching or germination rate. Some of the first observations are an increase of the mutation rate of embryonic lethals in the second generation of Arabidopsis seeds, somatic mutations, and a reduction of growth rates of corn plants and a reduction of life span of Artemia salina shrimps. The different passive detector systems are also in a good shape and give access to a proper dosimetric analysis. The results are summarized, and some aspects of future analysis are shown.