Microbial sterilization in ultra-high vacuum and outer space - A kinetic comparison
Reaction kinetics of microbial sterilization in ultrahigh vacuum and in outer space
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Reaction kinetics of microbial sterilization in ultrahigh vacuum and in outer space
Automated microbial metabolism life detection experiments for exobiological studies
Microbial assay of lunar core and surface samples obtained from Apollo 11 and 12 flights
Work undertaken to improve the estimation of the probability of release of microorganisms from unmanned Martian landing spacecraft is summarized. An analytical model is described for the development of numerical values for release parameters and release mechanisms applicable to flight missions are defined. Laboratory test data are used to evolve parameter values for use by flight projects in estimating numerical values for release probabilities. The analysis treats microbial burden located on spacecraft surfaces, between mated surfaces, and encapsulated within materials.
Experiments were performed to determine the influence closed microbial ecologies have on modification or simplification of natural intestinal flora of ferrets in a closed environmental system. On the basis of previous tests in which certain species (Salmonella and Bacteroides) were decreased at 90 days of enclosure, a second trial was constructed for 180-day tests. In this trial there was little difference in the 8 major classes of intestinal flora between animals in the Open and Closed environmental groups except for the level of Lactobacillus. It is of extreme importance to note that when both Open and Closed groups contracted hemorrhagic gastritis, the interrelationship of this agent with other intestinal flora produced a more profound effect on animals from the Closed Group, particularly with reference to Lactobacillus levels.
A single reproducible area of the preparation in a fixed focal plane is photographically scanned at intervals during incubation. The procedure can be used for evaluating the aerobic or anaerobic growth of many microbial cells simultaneously within a population. In addition, the microscope is not restricted to the viewing of any one microculture preparation, since the slide cultures are incubated separately from the microscope.
The Automated Microbial Metabolism Laboratory (AMML) 1971-1972 program involved the investigation of three separate life detection schemes. The first was a continued further development of the labeled release experiment. The possibility of chamber reuse without inbetween sterilization, to provide comparative biochemical information was tested. Findings show that individual substrates or concentrations of antimetabolites may be sequentially added to a single test chamber. The second detection system which was investigated for possible inclusion in the AMML package of assays, was nitrogen fixation as detected by acetylene reduction. Thirdly, a series of preliminary steps were taken to investigate the feasibility of detecting biopolymers in soil. A strategy for the safe return to Earth of a Mars sample prior to manned landings on Mars is outlined. The program assumes that the probability of indigenous life on Mars is unity and then broadly presents the procedures for acquisition and analysis of the Mars sample in a manner to satisfy the scientific community and the public that adequate safeguards are being taken.
Current U.S. planetary quarantine standards based on international agreements require consideration of the probability of contamination (Pc) of the outer planets, Venus, Jupiter, Saturn, etc. One of the key parameters in estimation of the Pc of these planets is the probability of growth (Pg) of terrestrial microorganisms on or near these planets. For example, Jupiter and Saturn appear to have an atmosphere in which some microbial species could metabolize and propagate. This study includes investigation of the likelihood of metabolism and propagation of microbes suspended in dynamic atmospheres. It is directed toward providing experimental information needed to aid in rational estimation of Pg for these outer plants.
Conference on Apollo 16 experiment for measuring microbial response to space environment
Exposure of test systems in space required the fabrication of specialized hardware termed a Microbial Ecology Evaluation Device that had individual test chambers and a complex optical filter system. The characteristics of this device and the manner in which it was deployed in space are described.
The fluid actinometry portion of the Microbial Response to Space Environment Experiment (M191) was designed for measurement of the solar energy that penetrates certain optical filter systems during exposure in space. Potassium ferrioxalate was used to measure energy at peak wavelengths of 254, 280, and 300 nanometers because of its high degree of sensitivity and its linear response to the middle ultraviolet regions.
Lithium fluoride thermoluminescent chips were used to provide an integrated dose from the broad spectrum of ionizing radiation to the Microbial Response to Space Environment Experiment (M191). The chips were positioned in the flight hardware to provide data on ionizing radiation within specific volume segments. A uniform radiation dose of 4.8 x 0.001 plus or minus 2 x 0.0001 joule/kg resulted.
Planetary quarantine analyses performed for recent unmanned Mars and Venus missions assumed that the probability of contamination by a spacecraft given accidental impact was equivalent to one. However, in the case of the gaseous outer planets, the heat generated during the inadvertent entry of a spacecraft into the planetary atmosphere might be sufficient to cause significant microbial burden reduction. This could affect navigation strategy by reducing the necessity for biasing the aim point away from the planets. An effort has been underway to develop the tools necessary to predict temperature histories for a typical spacecraft during inadvertent entry. In order that the results have general applicability, parametric analyses were performed. The thermal response of the spacecraft components and debris resulting from disintegration was determined. The temperature histories of small particles and composite materials, such as thermal blankets and an antenna, were given special attention. Guidelines are given to indicate the types of components and debris most likely to contain viable organisms, which could contaminate the lower layers of the Jovian atmosphere (approximately one atmosphere of pressure).
One of the key parameters in estimation of the probability of contamintion of the outer planets (Jupiter, Saturn, Uranus, etc.) is the probability of growth (Pg) of terrestrial microorganisms on or near these planets. For example, Jupiter appears to have an atmosphere in which some microbial species could metabolize and propagate. This study includes investigation of the likelihood of metabolism and propagation of microbes suspended in dynamic atmospheres. It is directed toward providing experimental information needed to aid in rational estimation of Pg for these outer planets. Current work is directed at demonstration of aerial metabolism under near optimal conditions and tests of propagation in simulated Jovian atmospheres.
Development of the automated microbial metabolism laboratory (AMML) concept is reported. The focus of effort of AMML was on the advanced labeled release experiment. Labeled substrates, inhibitors, and temperatures were investigated to establish a comparative biochemical profile. Profiles at three time intervals on soil and pure cultures of bacteria isolated from soil were prepared to establish a complete library. The development of a strategy for the return of a soil sample from Mars is also reported.
Luciferase assay for adenosine triphosphate (ATP) was optimized for pure bacteria in broth in order to evaluate if changes in bacterial ATP content could be used as a rapid measure of antibiotic effect on microorganisms. Broth cultures of log phase bacteria were incubated at 310 K (37 C) for 2.5 hours at antimicrobial concentrations which resulted in the best discrimination between sensitive and resistant strains. Eighty-seven strains of 11 bacterial species were studied for their susceptibility to 12 commonly used antimicrobial agents: ampicillin, Penicillin G, nafcillin, carbenicillin, cephalothin, tetracycline, erythromycin, clindamycin, gentamicin, nitrofurantoin, colistin, and chloramplenicol. The major advantage of the ATP system over existing methods of rapid microbial susceptibility testing is that the assay can be made specific for bacterial ATP.
Operational aspects associated with the experiment and the activities of medically important microorganisms recovered from the Apollo crewmen are evaluated. A variety of potential pathogens was recovered from each of the prime and backup crew members before and after flight. However, no disease events were reported. Candida albicans and Staphylococcus aureus were shown to be transferred from one crewmember to another during the flight. No other medically significant changes in the microbial population were observed.
Design analysis of a microbial load monitor system flight engineering model was presented. Checkout of the card taper and media pump system was fabricated as well as the final two incubating reading heads, the sample receiving and card loading device assembly, related sterility testing, and software. Progress in these areas was summarized.