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At least 37 records · Page 2

Titan Ice and Dust Experiment (TIDE): Detection and Analysis of Compounds of Interest to Astrobiology in the Lower Atmosphere and Surface of Titan

The Titan Orbiter Aerorover Mission (TOAM) is a proposed concept for the Solar System Exploration Visions Mission, Titan Explorer, a follow-on to the Cassini-Huygens mission. TOAM would use a Titan polar orbiter and a lighter-than-air aerorover to investigate the surface and atmosphere of Titan. Astrobiology issues will be addressed though TOAM investigations including, for example: Distribution and composition of organics (atmospheric, aerosol, surface); Organic chemical processes, their chemical context and energy sources; and Seasonal variations and interactions of the atmosphere and surface. The TIDE instrument will perform in-situ analyses to obtain comprehensive and sensitive molecular and elemental assays of volatile organics in the atmosphere, oceans and surface. TIDE chemical analyses are conducted by a Gas Chromatograph-Ion Mobility Spectrometer (GC-IMS). This TIDE GC-IMS was a component of the mini-Cometary Ice and Dust Experiment (mini-CIDEX) developed for the chemical analysis of a cometary environment. Both the GC and helium IMS of mini-CIDEX have been further developed to better meet the analytical and operational requirements of the TOAM. application. A Micro-ElectroMechanical System (MEMS) GC and Mini-Cell helium IMS are under development to replace their respective mini-CIDEX components, providing similar or advanced analytical capabilities.

Kojiro, Daniel R.↗

Standardization of Spore Inactivation Method for PMA-PhyloChip Analysis

In compliance with the Committee on Space Research (COSPAR) planetary protection policy, National Aeronautics and Space Administration (NASA) monitors the total microbial burden of spacecraft as a means for minimizing the inadvertent transfer of viable contaminant microorganisms to extraterrestrial environments (forward contamination). NASA standard assay-based counts are used both as a proxy for relative surface cleanliness and to estimate overall microbial burden as well as to assess whether forward planetary protection risk criteria are met for a given mission, which vary by the planetary body to be explored and whether or not life detection missions are present. Despite efforts to reduce presence of microorganisms from spacecraft prior to launch, microbes have been isolated from spacecraft and associated surfaces within the extreme conditions of clean room facilities using state of the art molecular technologies. Development of a more sensitive method that will better enumerate all viable microorganisms from spacecraft and associated surfaces could support future life detection missions. Current culture-based (NASA standard spore assay) and nucleic-acid-based polymerase chain reaction (PCR) methods have significant shortcomings in this type of analysis. The overall goal of this project is to evaluate and validate a new molecular method based on the use of a deoxyribonucleic acid (DNA) intercalating agent propidium monoazide (PMA). This is used in combination with DNA microarray (PhyloChip) which has been shown to identify very low levels of organisms on spacecraft associated surfaces. PMA can only penetrate the membrane of dead cells. Once penetrated, it intercalates the DNA and, upon photolysis using visible light it produces stable DNA monoadducts. This allows DNA to be unavailable for further PCR analysis. The specific aim of this study is to standardize the spore inactivation method for PMA-PhyloChip analysis. We have used the bacterial spores Bacillus subtilis 168 (standard laboratory isolate) as a test organism.

spore inactivation↗

Microbial Characterization Space Solid Wastes Treated with a Heat Melt Compactor

The on going purpose of the project efforts was to characterize and determine the fate of microorganisms in space-generated solid wastes before and after processing by candidate solid waste processing. For FY 11, the candidate technology that was assessed was the Heat Melt Compactor (HMC). The scope included five HMC. product disks produced at ARC from either simulated space-generated trash or from actual space trash, Volume F compartment wet waste, returned on STS 130. This project used conventional microbiological methods to detect and enumerate microorganisms in heat melt compaction (HMC) product disks as well as surface swab samples of the HMC hardware before and after operation. In addition, biological indicators were added to the STS trash prior to compaction in order to determine if these spore-forming bacteria could survive the HMC processing conditions, i.e., high temperature (160 C) over a long duration (3 hrs). To ensure that surface dwelling microbes did not contaminate HMC product disk interiors, the disk surfaces were sanitized with 70% alcohol. Microbiological assays were run before and after sanitization and found that sanitization greatly reduced the number of identified isolates but did not totally eliminate them. To characterize the interior of the disks, ten 1.25 cm diameter core samples were aseptically obtained for each disk. These were run through the microbial characterization analyses. Low counts of bacteria, on the order of 5 to 50 per core, were found, indicating that the HMC operating conditions might not be sufficient for waste sterilization. However, the direct counts were 6 to 8 orders of magnitude greater, indicating that the vast majority of microbes present in the wastes were dead or non-cultivable. An additional indication that the HMC was sterilizing the wastes was the results from the added commercial spore test strips to the wastes prior to HMC operation. Nearly all could be recovered from the HMC disks post-operation and all were showed negative growth when run through the manufacturer's protocol, meaning that the 106 or so spores impregnated into the strips were dead. Control test strips, i.e., not exposed to the HMC conditions were all strongly positive. One area of concern is that the identities of isolates from the cultivable counts included several human pathogens, namely Staphylococcus aureus. The project reported here provides microbial characterization support to the Waste Management Systems element of the Life Support and Habitation Systems program.

Strayer, Richard F.↗

Cytometer on a chip

An assay technique for label-free, highly parallel, qualitative and quantitative detection of specific cell populations in a sample and for assessing cell functional status, cell-cell interactions and cellular responses to drugs, environmental toxins, bacteria, viruses and other factors that may affect cell function. The technique includes a) creating a first array of binding regions in a predetermined spatial pattern on a sensor surface capable of specifically binding the cells to be assayed; b) creating a second set of binding regions in specific spatial patterns relative to the first set designed to efficiently capture potential secreted or released products from cells captured on the first set of binding regions; c) contacting the sensor surface with the sample, and d) simultaneously monitoring the optical properties of all the binding regions of the sensor surface to determine the presence and concentration of specific cell populations in the sample and their functional status by detecting released or secreted bioproducts.

Lynes, Michael A.↗

Organic Biomarker-Based Assays to Evaluate Total Bioburden and Organic Compounds on Space Flight Hardware

Meeting planetary protection (PP) requirements for space flight hardware may involve bioburden reduction by dry heat microbial reduction (DHMR). The NASA standard assay to demonstrate the reduction of organisms involves the swabbing of surfaces, heat shock of the extracted samples, plating of the samples on Trypticase Soy Agar (TSA), and counting colony forming units after an incubation period. The standard assay uses enumeration of heat tolerant spore-formers as a proxy for total bioburden and is generally expected to provide a lower limit. We suggest that a better estimate of the total bioburden could be obtained through sampling and analysis of organic biomarkers. As biological organisms are fundamentally organic in chemistry (i.e. carbon containing materials) it is important to characterize the biomarker compounds that are released from organisms that 1) exist on flight hardware before microbial reduction and 2) left behind from the killed organisms following microbial reduction.

Locke, Darren R.↗

Electrochemical Life Detection Methods for Ocean World Exploration

Ubiquitous across terrestrial life is cellular machinery that allows chemical energy flow by facilitating and regulating electron-transfer and chemical modification pathways. Key classes of energy transport molecules enable this movement of electrons for a variety of biological purposes. Additionally, biological enzymes function to add or remove functional groups such asphosphate moieties to redox biomolecules. Presumably, extraterrestrial life is likely to rely on similar energy transport mechanisms. With the search for life in our solar system focused on the icy satellites of Jupiter and Saturn, Europa and Enceladus, developing instrumentation capable of measuring electrochemical redox signatures representative of biomolecules or enzymatic activity in seawater appears a promising and novel means of life detection. Here, we report our adaptation of the Mars Phoenix Wet Chemistry Laboratory (WCL) electroanalytical voltammetry capabilities to assay life-critical redox molecules in synthetic seawater representative of a saline alkaline solution similar to what has been predicted from the Cassini mission data of Enceladus’ sub-surface ocean. In addition, we employ a well-established electrochemical assay that indicates phosphatase activity by comparing substrate and product redox signatures. Our study demonstrated a 10 nM limit of detection for biological redox molecules and a 3 aM limit of detection for alkaline phosphatase in seawater. Incorporation of these methods into next generation WCL payloads aimed at ocean world life detection will enable the search for biological redox-active species and enzymatic activity as indicators of life.

planetary instruments↗

Novel Confocal Microscopic and Flow Cytometric Based Assays to Visualize and Detect the (Beta)2-Adrenergic Receptor in Human Lymphocyte and Mononuclear Cell Populations

The data show that immunophenotyping of leukocyte populations with (beta)2AR is possible with the commercially available Ab, although the FC assay is limited to the IST as a result of the Ab binding site to the intracellular C-terminus of the 2AR. The FC assay has applications for measuring alterations in total (beta)2AR in human leukocyte populations as changes in fluorescence. In addition, CM confirms that both surface and intracellular compartments stain positively for the (beta)2AR and can be used for qualitative assays that screen for changes in receptor compartmentalization and localization.

Salicru, A. N.↗

Heat transfer effects of longitudinal vortices embedded in a turbulent boundary layer

The results of tests with five different vortex generators in an open circuit wind tunnel are reported. The tests were performed to enhance the data base on heat transfer effects of an isolated longitudinal vortex in a turbulent boundary layer and the effects of embedded vortex pairs. A heated plate was set in the boundary layer downstream of a tripping mechanism and measurements were made of the free stream and surface temperatures using thermocouples. A four-hole cobra probe assayed the velocity profiles. Trials revealed that vortex pairs with scales larger than the boundary layer thickness induce significant changes in the Stanton number. A 15 percent Stanton number increase was produced by isolated vortices, which swept cool free stream fluid into the near-wall region.

Eibeck, P. A.↗

Enhanced Characterization of Microorganisms in the Spacecraft Environment

Spacecraft such as the International Space Station (ISS) and the space shuttles are enclosed environments where crewmembers may spend long periods of time. Currently, crewmembers spend approximately a period of 6 months in the ISS. It is known that these prolonged stays in space may result in weakening of the immune system. Therefore, exposure to opportunistic pathogens or high concentrations of environmental microorganisms may compromise the health of the crew. The detection of biocontaminants in spacecraft environments utilizes culture-based methodology, omitting greater than 90% of all microorganisms including pathogens such as Legionella and Cryptosporidium. Culturable bacteria and fungi have been the only allergens studied; the more potent allergens, such as those from dust mites, have never been tested for in spacecraft environments. In addition, no attempts have been made to monitor microbial toxins in spacecrafts. The present study utilized quantitative polymerase chain reaction (QPCR) as a novel approach for monitoring microorganisms in the spacecraft environment. QPCR is a molecular biology technique that does not rely on the physiological state of the organisms for identification, thereby enabling detection of both culturable and non-culturable organisms. In this project, specific molecular primers and probes were utilized for the detection and quantitation of two fungi of concern in indoor environments, Aspergillus fumigatus and Stachybotrys chartarum. These organisms were selected because of the availability of PCR primers and probes, and to establish the sample processing and analysis methodology that may be employed with additional organisms. Purification methods and QPCR assays were optimized for the detection of these organisms in air, surface, and water; and sample processing and analysis protocols were developed. Preliminary validation of these protocols was conducted in the laboratory with air, surface, and water samples seeded with known concentrations of the target organisms. Additional studies were conducted with bulk materials (HEPA filter pleats and particulate found on the filter screen) obtained from the ISS.

Cruz, Patricia↗

Detection of Antimicrobial Resistance Genes Associated With the International Space Station Environmental Surfaces

Antimicrobial resistance (AMR) is a global health issue. In an effort to minimize this threat to astronauts, who may be immunocompromised and thus at a greater risk of infection from antimicrobial resistant pathogens, a comprehensive study of the ISS “resistome’ was conducted. Using whole genome sequencing (WGS) and disc diffusion antibiotic resistance assays, 9 biosafety level 2 organisms isolated from the ISS were assessed for their antibiotic resistance. Molecular analysis of AMR genes from 24 surface samples collected from the ISS during 3 different sampling events over a span of a year were analyzed with Ion AmpliSeq™ and metagenomics. Disc diffusion assays showed that Enterobacter bugandensis strains were resistant to all 9 antibiotics tested and Staphylococcus haemolyticus being resistant to none. Ion AmpliSeq™ revealed that 123 AMR genes were found, with those responsible for beta-lactam and trimethoprim resistance being the most abundant and widespread. Using a variety of methods, the genes involved in antimicrobial resistance have been examined for the first time from the ISS. This information could lead to mitigation strategies to maintain astronaut health during long duration space missions when return to Earth for treatment is not possible.

Antimicrobial resistance↗

Venus Orbital Imaging Radar mission analysis

Mission objectives for the Venus Orbital Imaging Radar (VOIR) project are outlined with attention to its scientific instrumentation. Design parameters of the SAR (Synthetic Aperture Radar) are described, including a high resolution capability of 200 m from a circular orbit of 375 km, which will be able to map the entire surface of the planet. Nineteen atmospheric experiments are foreseen, among them: CO2 stability assays, measurements of vertical mass transfer rates, observations of cloud circulation and composition, and an evaluation of solar wind effects on atmospheric dynamics. Navigation and orbital injection plans are reviewed, noting that the 1983 launch window (using STS) is optimum.

Nock, K. T.↗

Automated life-detection experiments for the Viking mission to Mars

As part of the Viking mission to Mars in 1975, an automated set of instruments is being built to test for the presence of metabolizing organisms on that planet. Three separate modules are combined in this instrument so that samples of the Martian surface can be subjected to a broad array of experimental conditions so as to measure biological activity. The first, the Pyrolytic Release Module, will expose surface samples to a mixture of C-14O and C-14O2 in the presence of Martian atmosphere and a light source that simulates the Martian visible spectrum. The assay system is designed to determine the extent of assimilation of CO or CO2 into organic compounds. The Gas Exchange Module will incubate surface samples in a humidified CO2 atmosphere. At specified times, portions of the incubation atmosphere will be analyzed by gas chromatography to detect the release or uptake of CO2 and several additional gases. The Label Release Module will incubate surface samples with a dilute aqueous solution of simple radioactive organic substrates in Martian atmosphere, and the gas phase will be monitored continuously for the release of labeled CO2.

Klein, H. P.↗

Zoledronate Prevents Simulated Weightlessness-Induced Bone Loss in the Cancellous Compartment While Blunting the Efficacy of a Mechanical Loading Countermeasure

Astronauts using high-force resistance training while weightless show a high-turnover remodeling state within the skeletal system, with resorption and formation biomarkers being elevated. One countermeasure for the skeletal health of astronauts includes an antiresorptive of the bisphosphonate (BP) drug class. We asked, does the combination of an anti-resorptive and high-force exercise during weightlessness have negative effects on bone remodeling and strength? In this study, we developed an integrated model to mimic the mechanical strain of exercise via cyclical loading (CL) in mice treated with the BP Zoledronate (ZOL) combined with hind limb unloading (HU) to simulate weightlessness. We hypothesized that ZOL prevents structural degradation from simulated weightlessness and that CL and ZOL interact to render CL less effective. Thirty-two C57BL/6 mice (male, 16 weeks old, n=8/group) were exposed to 3 weeks of either HU or normal ambulation (NA). Cohorts of mice received one subcutaneous injection of ZOL (45μg/kg), or saline vehicle (VEH), prior to the start of HU. The right tibia was axially compressed in vivo 60x/day to 9N (+1200μstrain on the periosteal surface) and repeated 3x/week during HU. Left tibiae served as a within subject, non-compressed control. Ex vivo μCT was performed on all subjects to determine cancellous and cortical architectural parameters. Static and dynamic histomorphometry were carried out for the left and right tibiae to determine osteoclast- and osteoblast relevant surfaces. Further, micro damage was assessed in select groups by basic-fuchsin staining to test whether CL had an effect. For all assays, a multivariate (2x2x2) ANCOVA model was used to account for body weight changes. Additionally, for the tibiae, we incorporated a random effect for the subject (hence, a mixed model) to account for observations of both left and right tibiae within each subject. P < 0.05 was considered significant. In the cancellous compartment of the proximal tibial metaphysis, we observed a main effect from each independent variable, as determined by structural and histomorphometric assays. Specifically, as expected, ZOL showed an increase in the cancellous bone volume to total volume fraction (BV/TV, +32%) and trabecular number (+18%) compared to the VEH. As expected, ZOL decreased osteoclast surface (OC/BS) by -45% compared to VEH. Surprisingly, ZOL reduced mineralizing surface (MS/BS) and bone formation rate (BFR), indicators of osteoblast activity, by -40% and -54%, respectively, compared to VEH. Altogether, ZOL-treated mice displayed a low turnover state of remodeling in the metaphysis. In the context of skeletal aging, we speculate that ZOL prevented age-related cancellous strut loss during the experiment. As a main effect, as expected, HU decreased BV/TV by - 31% via reductions in both trabecular thickness (-11%) and number (-22%) compared to NA controls. Additionally, HU decreased MS/BS by -38% and bone formation rate (BFR) by -50% compared to NA controls. Altogether, these data are consistent with structural degradation resulting from imbalanced remodeling that favors resorption. As a main effect, CL increased BV/TV by +15% via increased trabecular thickness (+12%) compared to the noncompressed limb. As expected, CL increased MS/BS (+20%) and BFR (+24%), indicating osteoblast mineralization contributed to bone gains. These data show that CL provided an anabolic stimulus to the cancellous tissue. We observed unique interactions in ZOL*CL and HU*CL. First, ZOL prevented CL-induced increases in BV/TV and trabecular number, as compared to VEH. In the context of skeletal aging, these data suggest no added benefit from CL in the ZOL-treated mice. Interestingly, no microdamage was observed in mice that were unloaded and treated with ZOL (independent of CL). Secondly, HU prevented CL-induced increases in BFR, as compared to NA controls. These data suggest that either exercise is less effective or the kinetics of formation are slower during simulated weightlessness. Osteoclast surface was unchanged by either treatment. Thus, in contrast to exercising astronauts, these data do not suggest a high-turnover state in the metaphysis. To assess mechanical properties as a function of HU or ZOL, we tested the left femur in three-point bending ex vivo. As expected, HU decreased stiffness (-30%) compared to NA, and ZOL increased stiffness compared to VEH (+28%). Interestingly, HU increased the post-yield displacement, related to collagenous tensile loading, compared to NA (+20%). ZOL increased yield force (+11%) and ultimate force (+17%), which seems to explain the significant effect of ZOL increasing total energy (work-to-fracture, +15%), while not affecting the post yield displacement. Taken together, ZOL did not have detrimental affect on mechanical properties. Our integrated model simulates the combination of weightlessness, exercise-induced mechanical strain, and anti-resorptive treatment that astronauts experience during space missions. We conclude that Zoledronate was an effective countermeasure against weightlessness-induced bone loss, though zoledronate, as well as weightlessness, rendered exercise-related mechanical loading less effective.

bone↗

Abiotic Versus Biotic Weathering Of Olivine As Possible Biosignatures

We are investigating the weathering of silicate minerals by both purely inorganic, and biologically mediated processes using field-emission scanning electron microscopy (FESEM) and energy dispersive x-ray spectroscopy (EDS). By resolving surface textures and chemical compositions of weathered surfaces at the sub-micron scale we hope to be able to distinguish abiotic from biotic weathering processes and so establish a new biosignature applicable to the study of astromaterials including but not limited to the Martian meteorites. Sterilized olivine grains (San Carlos, Arizona) no more than 1-2 mm in their longest dimension were optically assayed to be uniform in color and free of inclusions were selected as weathering subjects. Prior to all experiments surface morphologies and Fe/Mg ratios were determined for each grain using FE-SEM and EDS. Experiments were divided into two categories abiotic and biotic and were compared with "naturally" weathered samples. For the preliminary experiments, two trials (open and closed to the ambient laboratory environment) were performed under abiotic conditions, and three trials under biotic conditions (control, day 1 and day 2). The open system abiotic trials used sterile grains heated at 98 C and 200 C for both 24 and 48 hours in 1L double distilled de-ionized water. The closed system abiotic trials were conducted under the same conditions but in a sealed two layer steel/Teflon "bomb" apparatus. The biotic trials used sterile grains mounted in a flow-through device attached to a wellhead on the Columbia River aquifer. Several discolored, altered, grains were selected to document "natural" weathering surface textures for comparison with the experimental samples. Preliminary results indicate there are qualitative differences in weathered surface textures among all the designed experiments. The olivine grains in abiotic trials displayed etching, pitting, denticulate margins, dissolution and clay formation. The scale of the features ranged from tens to a few microns with textures that remained relatively sharp and were crystallographically controlled. These results were comparable to that observed in the "naturally" weathered comparison/reference grains. Chemical analysis by EDS indicates these textures correlated with the relative loss of Mg and Fe cations by diffusional processes. In contrast the biotic results indicated changes in the etching patterns on the scale of hundreds of nm, which are neither sharp nor crystallographically controlled (nanoetching). Organisms, organic debris and/or extracellular polymeric substances (biofilm) were often in close proximity or direct contact with the nanoetching. While there are many poorly constrained variables in natural weathering experiments to contend with, such as the time scale, the chemistry of the fluids and degree of biologic participation, some preliminary observations can be made: (1) certain distinct surface textures appear correlated with the specific processes giving rise to these textures; (2) the process of diffusing cations can produce many similar styles of surface textural changes; and (3) the main difference between abiotic and biotically produced weathering is the scale (microns versus nanometers) and the style (crystallographically versus noncrystallographically controlled) of the textural features. Further investigation into nanosize scale surface textures should attempt to quantify both textures and chemical changes of the role of microorganisms in the weathering of silicates. Additional experiments addressing nanoscale textures of shock features for comparison with the current data set.

Longazo, Teresa G.↗

Wipe-rinse technique for quantitating microbial contamination on large surfaces

The evaluation of an improved wipe-rinse technique for the bioassay of large areas was undertaken due to inherent inadequacies in the cotton swab-rinse technique to which assay of spacecraft is currently restricted. Four types of contamination control cloths were initially tested. A polyester-bonded cloth (PBC) was selected for further evaluation because of its superior efficiency and handling characteristics. Results from comparative tests with PBC and cotton swabs on simulated spacecraft surfaces indicated a significantly higher recovery efficiency for the PBC than for the cotton (90.4 versus 75.2%). Of the sampling area sites studied, PBC was found to be most effective on surface areas not exceeding 0.74 sq m (8.0 sq ft).

Kirschner, L. E.↗

Twisting integrin receptors increases endothelin-1 gene expression in endothelial cells

A magnetic twisting stimulator was developed based on the previously published technique of magnetic twisting cytometry. Using ligand-coated ferromagnetic microbeads, this device can apply mechanical stresses with varying amplitudes, duration, frequencies, and waveforms to specific cell surface receptors. Biochemical and biological responses of the cells to the mechanical stimulation can be assayed. Twisting integrin receptors with RGD (Arg-Gly-Asp)-containing peptide-coated beads increased endothelin-1 (ET-1) gene expression by >100%. In contrast, twisting scavenger receptors with acetylated low-density lipoprotein-coated beads or twisting HLA antigen with anti-HLA antibody-coated beads did not lead to alterations in ET-1 gene expression. In situ hybridization showed that the increase in ET-1 mRNA was localized in the cells that were stressed with the RGD-coated beads. Blocking stretch-activated ion channels with gadolinium, chelating Ca2+ with EGTA, or inhibiting tyrosine phosphorylation with genistein abolished twist-induced ET-1 mRNA elevation. Abolishing cytoskeletal tension with an inhibitor of the myosin ATPase, with an inhibitor of myosin light chain kinase, or with an actin microfilament disrupter blocked twisted-induced increases in ET-1 expression. Our results are consistent with the hypothesis that the molecular structural linkage of integrin-cytoskeleton is an important pathway for stress-induced ET-1 gene expression.

Non-NASA Center↗

Microbial Characterization of Solid-Wastes Treated with Heat Melt Compaction Technology

The research purpose of the project was to determine the fate of microorganisms in space-generated solid wastes after processing by a Heat Melt Compactor (HMC), which is a candidate solid waste treatment technology. Five HMC product disks were generated at Ames Research Center (ARC), Waste Management Systems element. The feed for two was simulated space-generated trash and feed for three was Volume F compartment wet waste returned on STS 130. Conventional microbiological methods were used to detect and enumerate microorganisms in HMC disks and in surface swab samples of HMC hardware before and after operation. Also, biological indicator test strips were added to the STS trash prior to compaction to test if HMC processing conditions, 150 C for approx 3 hr and dehydration, were sufficient to eliminate the test bacteria on the strips. During sample acquisition at KSC, the HMC disk surfaces were sanitized with 70% alcohol to prevent contamination of disk interiors. Results from microbiological assays indicated that numbers of microbes were greatly reduced but not eliminated by the 70% alcohol. Ten 1.25 cm diameter cores were aseptically cut from each disk to sample the disk interior. The core material was run through the microbial characterization analyses after dispersal in sterile diluent. Low counts of viable bacteria (5 to 50 per core) were found but total direct counts were 6 to 8 orders of magnitude greater. These results indicate that the HMC operating conditions might not be sufficient for complete waste sterilization, but the vast majority of microbes present in the wastes were dead or non-cultivable after HMC treatment. The results obtained from analyses of the commercial spore test strips that had been added fo the wastes prior to HMC operation further indicated that the HMC was sterilizing the wastes. Nearly all strips were recovered from the HMC disks and all of these were negative for spore growth when run through the manufacturer's protocol. The 10(exp 6) or so spores impregnated into the strips were no longer viable. Control test strips, i.e., not exposed to the HMC conditions, were all strongly positive. All isolates from the cultivable counts were identified, leading to one concern: several were identified as Staphylococcus aureus, a human pathogen. The project reported here provides microbial characterization support to the Waste Management Systems element of the Life Support and Habitation Systems program.

Strayer, Richard F.↗