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NASA NTRS1995

PCR Based Microbial Monitor for Analysis of Recycled Water Aboard the ISSA: Issues and Prospects

The monitoring of spacecraft life support systems for the presence of health threatening microorganisms is paramount for crew well being and successful completion of missions. Development of technology to monitor spacecraft recycled water based on detection and identification of the genetic material of contaminating microorganisms and viruses would be a substantial improvement over current NASA plans to monitor recycled water samples that call for the use of conventional microbiology techniques which are slow, insensitive, and labor intensive. The union of the molecular biology techniques of DNA probe hybridization and polymerase chain reaction (PCR) offers a powerful method for the detection, identification, and quantification of microorganisms and viruses. This technology is theoretically capable of assaying samples in as little as two hours with specificity and sensitivity unmatched by any other method. A major advance in probe-hybridization/PCR has come about in a technology called TaqMan(TM), which was invented by Perkin Elmer. Instrumentation using TaqMan concepts is evolving towards devices that could meet NASA's needs of size, low power use, and simplicity of operation. The chemistry and molecular biology needed to utilize these probe-hybridization/PCR instruments must evolve in parallel with the hardware. The following issues of chemistry and biology must be addressed in developing a monitor: Early in the development of a PCR-based microbial monitor it will be necessary to decide how many and which organisms does the system need the capacity to detect. We propose a set of 17 different tests that would detect groups of bacteria and fungus, as well as specific eukaryotic parasites and viruses; In order to use the great sensitivity of PCR it will be necessary to concentrate water samples using filtration. If a lower limit of detection of 1 microorganism per 100 ml is required then the microbes in a 100 ml sample must be concentrated into a volume that can be added to a PCR assay; There are not likely to be contaminants in ISSA recycled water that would inhibit PCR resulting in false-negative results; The TaqMan PCR product detection system is the most promising method for developing a rapid, highly automated gene-based microbial monitoring system. The method is inherently quantitative. NASA and other government agencies have invested in other technologies that, although potentially could lead to revolutionary advances, are not likely to mature in the next 5 years into working systems; PCR-based methods cannot distinguish between DNA or RNA of a viable microorganism and that of a non-viable organism. This may or may not be an important issue with reclaimed water on the ISSA. The recycling system probably damages the capacity of the genetic material of any bacteria or viruses killed during processing to serve as a template in a PCR desinged to amplify a large segment of DNA (less than 650 base pairs). If necessary, vital dye staining could be used in addition to PCR, to enumerate the viable cells in a water sample; The quality control methods have been developed to insure that PCR's are working properly, and that reactions are not contaminated with PCR carryover products which could lead to the generation of false-positive results; and The sequences of the small rRNA subunit gene for a large number of microorganisms are known, and they consititue the best database for rational development of the oligonucleotide reagents that give PCR its great specificity. From those gene sequences, sets of oligonucleotide primers for PCR and Taqman detection that could be used in a NASA microbial monitor were constructed using computer based methods. In addition to space utilization, a microbial monitior will have tremendous terrestrial applications. Analysis of patient samples for microbial pathogens, testing industrial effluent for biofouling bacteria, and detection biological warfare agents on the battlefield are but a few of the diverse potential uses for this technology. Once fully developed, gene-based microbial monitors will become the fundamental tool in every lab that tests for microbial contaminants, and serve as a powerful weapon in mankind's war with the germ world.

Cassell, Gail H.↗

NASA NTRS1997

Development of an Antimicrobial Susceptibility Testing Method Suitable for Performing During Space Flight

Very little is known regarding the affects of the microgravity environment of space flight upon the action of antimicrobial agents on bacterial pathogens. This study was undertaken to develop a simple method for conducting antibacterial susceptibility tests during a Space Shuttle mission. Specially prepared susceptibility test research cards (bioMerieux Vitek, Hazelwood, MO) were designed to include 6-11 serial two-fold dilutions of 14 antimicrobial agents, including penicillins, cephalosporins, a Beta-lactamase inhibitor, vancomycin, erythromycin, tetracycline, gentamicin, ciprofloxacin, and trimethoprim/sulfamethoxazole. Minimal inhibitory concentrations (MICS) of the drugs were determined by visual reading of color endpoints in the Vitek research cards made possible by incorporation of a colorimetric growth indicator (alamarBlue(Trademark), Accumed International, Westlake, OH). This study has demonstrated reproducible susceptibility results when testing isolates of Staphylococcus aurezis, Group A Streptococcus, Enterococcusfaecalis, Escherichia coli (beta-lactamase positive and negative strains), Klebsiella pneumoniae, Enterobacter cloacae, and Pseudomoiias aeruginosa. In some instances, the MICs were comparable to those determined using a standard broth microdilution method, while in some cases the unique test media and format yielded slightly different values, that were themselves reproducible. The proposed in-flight experiment will include inoculation of the Vitek cards on the ground prior to launch of the Space Shuttle, storage of inoculated cards at refrigeration temperature aboard the Space Shuttle until experiment initiation, then incubation of the cards for 18-48 h prior to visual interpretation of MICs by the mission's astronauts. Ground-based studies have shown reproducible MICs following storage of inoculated cards for 7 days at 4-8 C to accommodate the mission's time schedule and the astronauts' activities. For comparison, ground-based control (normal gravity) MIC values will be generated by simultaneous inoculation and incubation of a second set of test cards in a laboratory at the launch site. This procedure can provide a safe and compact experiment that should yield new information on the affects of microgravity on the biological activities of various classes of antibiotics.

Jorgensen, James H.↗

NASA NTRS1994

Land Use Patterns and Fecal Contamination of Coastal Waters in Western Puerto Rico

The Department of Environmental Health of the Graduate School of Public Health of the Medical Sciences Campus, University of Puerto Rico (UPR-RCM) conducted this research project on how different patterns of land use affect the microbiological quality of rivers flowing into Mayaguez Bay in Western Puerto Rico. Coastal shellfish growing areas, stream and ocean bathing beaches, and pristine marine sites in the Bay are affected by the discharge of the three study rivers. Satellite imagery was used to study watershed land uses which serve as point and nonpoint sources of pathogens affecting stream and coastal water users. The study rivers drain watersheds of different size and type of human activity (including different human waste treatment and disposal facilities). Land use and land cover in the study watersheds were interpreted, classified and mapped using remotely sensed images from NASA's Landsat Thematic Mapper (TM). This study found there is a significant relationship between watershed land cover and microbiological water quality of rivers flowing into Mayaguez Bay in Western Puerto Rico. Land covers in the Guanajibo, Anasco, and Yaguez watersheds were classified into forested areas, pastures, agricultural zones and urban areas so as to determine relative contributions to fecal water contamination. The land cover classification was made processing TM images with IDRISI and ERDAS software.

Norat, Jose↗

NASA NTRS1999

Starch Metabolism in Space-Grown Soybean Seedlings

The research conducted during this grant is described. There were three major areas of study: These were: (1) the interaction of gravity and sugar metabolism in soybean; (2) the effects of gravity on the photosynthetic activity of Brassica rapa; (3) investigation as to the effects of microgravity on the interaction of a fungal root pathogen with soybean.

Guikema, James A.↗

NASA NTRS1999

Latent Viruses: A Space Travel Hazard??

A major issue associated with long-duration space flight is the possibility of infectious disease causing an unacceptable medical risk to crew members. Our proposal is designed to gain information that addresses several issues outlined in the Immunology/Infectious disease critical path. The major hypothesis addressed is that space flight causes alterations in the immune system that may allow latent viruses which are endogenous in the human population to reactivate and shed to higher levels than normal which can affect the health of crew members during a long term space-flight mission. We will initially focus our studies on the human herpesviruses and human polyomaviruses which are important pathogens known to establish latent infections in the human population. Both primary infection and reactivation from latent infection with this group of viruses can cause a variety of illnesses that result in morbidity and occasionally mortality of infected individuals. Effective vaccines exist for only one of the eight known human herpesviruses and the vaccine itself can still reactivate from latent infection. Available antivirals are of limited use and are effective against only a few of the human herpesviruses. Although most individuals display little if any clinical consequences from latent infection, events which alter immune function such as immunosuppressive therapy following solid organ transplantation are known to increase the risk of developing complications as a result of latent virus reactivation. This proposal will measure both the frequency and magnitude of viral shedding and genome loads in the blood from humans participating in activities that serve as ground based models of space flight conditions. Our initial goal is to develop sensitive quantitative competitive PCR- based assays (QC-PCR) to detect the herpesvirus Epstein-Barr virus (EBV), and the polyomaviruses SV40, BKV, and JCV. Using these assays we will establish baseline patterns of viral genome load in the blood and viral shedding from normal volunteers in a longitudinal study over I year in length. As a comparison, we will measure patterns of viral genome loads and shedding from individuals who are severely immunosuppressed, in whom herpesvirus reactivation or primary infection with a herpesvirus is known to cause complications. In addition, we will proceed to testing ground based analogs in collaboration with Dr. Duane Pierson (Lyndon B. Johnson Space Center). This will include measuring samples obtained from individuals living and working in the extreme environment of Antarctica. We expect to detect viral shedding or reactivation from most of the test groups, although the magnitude of shedding or reactivation cannot be predicted. The data accumulated from studies in this proposal should allow us to evaluate whether events that simulate certain aspects of space flight reactivate viral infections severe enough in nature that they may compromise the success of long-term space flight missions. These studies will also provide a foundation to monitor viral reactivation and shedding from crew members participating in actual space flight missions. We will present data showing the establishment of our QC-PCR assay for detection of EBV.

Ling, P. D.↗

NASA NTRS1999

Mars Environmental Compatibility Assessment (MECA): Identifying the Hazards of the Martian Soil

Sometime in the next decade NASA will decide whether to send a human expedition to explore the planet Mars. The Mars Environmental Compatibility Assessment (MECA) has been selected by NASA to evaluate the Martian environment for soil and dust hazards to human exploration. The integrated MECA payload contains three elements: a wet-chemistry laboratory, a microscopy station, and enhancements to a lander robot-arm system incorporating arrays of material patches and an electrometer to identify triboelectric charging during soil excavation. The wet-chemistry laboratory will evaluate samples of Martian soil in water to determine the total dissolved solids, redox potential, pH, and quantify the concentration of many soluble ions using ion-selective electrodes. These electrodes can detect potentially dangerous heavy-metal ions, emitted pathogenic gases, and the soil's corrosive potential. MECA's microscopy station combines optical and atomic-force microscopy with a robot-arm camera to provide imaging over nine orders of magnitude, from meters to nanometers. Soil particle properties including size, shape, color, hardness, adhesive potential (electrostatic and magnetic), will be determined on the microscope stage using an ar-ray of sample receptacles and collection substrates, and an abrasion tool,. The simple, rugged atomic-force microscope will image in the submicron size range and has the capability of performing a particle-by-particle analysis of the dust and soil. Although selected by NASA's Human Exploration and Development of Space Enterprise, the MECA instrument suite also has the capability to address basic geology, paleoclimate, and exobiology issues. To understand both contemporaneous and ancient processes on Mars, the mineralogical, petrological, and reactivity of Martian surface materials should be constrained: the NMCA experiment will shed light on these quantities through its combination of chemistry and microscopy. On Earth, the earliest forms of life are preserved as microfossils. The atomic-force microscope will have the required resolution to image down to the scale of terrestrial microfossils and beyond.

Meloy, T. P.↗

NASA NTRS1999

The MSP 2001 Mars Environmental Compatibility Assessment (MECA)

The Mars Environmental Compatibility Assessment (MECA) will evaluate the Martian environment for soil and dust-related hazards to human exploration as part of the Mars Surveyor Program 2001 Lander. Sponsored by the Human Exploration and Development of Space (HEDS) enterprise, MECA's goal is to evaluate potential geochemical and environmental hazards that may confront future Martian explorers, and to guide HEDS scientists in the development of high fidelity Mars soil simulants. The integrated MECA payload contains a wet-chemistry laboratory, a microscopy station, an electrometer to characterize the electrostatics of the soil and its environment, and arrays of material patches to study the abrasive and adhesive properties of soil grains. The instrument will acquire soil samples with a robotic arm equipped with a camera. MECA will examine surface and subsurface soil and dust in order to characterize particle size, shape, hardness, and also physical characteristics that may provide clues to mineralogy. MECA will characterize soil/water mixtures with respect to pH, redox potential, total dissolved ions, and trace toxins. MECA will determine the nature of electrostatic charging associated with excavation of soil, and the influence of ionizing radiation on material properties. It will also observe natural dust accumulation on engineering materials. To accomplish these objectives, MECA is allocated a mass of 10 kg within an enclosure of 35 x 25 x 15 cm. The Wet Chemistry Laboratory (WCL) consists of four identical cells that will accept samples from surface and subsurface regions accessible to the Lander's robotic arm, mix them with water, and perform extensive analysis of the solution. Ion-selective electrodes and related sensors will evaluate total dissolved solids, redox potential, pH, and the concentration of many soluble ions and gases in wet Martian soil. These electrodes can detect potentially dangerous heavy-metal ions, emitted pathogenic gases, and the soil's corrosive potential. Experiments will include cyclic voltammetry and anodic stripping voltammetry. Complementary to the Viking experiments, the chemical laboratory will characterize the water-soil solution rather than emitted gases. Nonetheless, through analysis of dissolved gases it will be able to replicate many of the Viking observations related to oxidants. MECA's microscopy station combines optical and atomic-force microscopy (AFM) in an actively focused, controlled illumination environment to image particles from millimeters to nanometers in size. Careful selection of substrates allows controlled experiments in adhesion, abrasion, hardness, aggregation, magnetic and other properties. Special tools allow primitive manipulation (brushing and scraping) of samples. Soil particle properties including size, shape, color, hardness, adhesive potential (electrostatic and magnetic), will be determined using an array of sample receptacles and collection substrates. The simple, rugged atomic-force microscope will image in the submicron size range and has the capability of performing a particle-by-particle analysis of the dust and soil. On Earth, the earliest forms of life are preserved as microfossils. The atomic-force microscope will have the required resolution to image down to the scale of terrestrial microfossils and beyond. Mounted on the end of the robot arm, MECA's electrometer actually consists of four types of sensors: an electric field meter, several triboelectricity monitors, an ion gauge, and a thermometer. Tempered only by ultraviolet-light-induced ions and a low-voltage breakdown threshold, the dry, cold, dusty martian environment presents an imposing electrostatic hazard to both robots and humans. The field meter will measure the ambient field on nearby objects while the triboelectric sensors, using identical circuitry, will measure the charge accumulated on test substances as they are dragged through the soil by the arm. The ion chamber, open to the environment, will sense both charged dust and free ions in the air. Over and above the potential threat to electronics, the electrostatic environment holds one of the keys to transport of dust and, consequently, Martian meteorology. Viewed with the robot arm camera, the abrasion and adhesion plates are strategically placed to allow direct observation of the interaction between materials and soils on a macroscopic scale. Materials of graded hardness are placed directly under the robot arm scoop to sense wear and soil hardness. A second array, placed on the lander deck, is deployed after the dust plume of landing has settled. It can be manipulated in a primitive fashion by the arm, first having dirt deposited on it from the scoop and subsequently shaken clean. A third array will passively collect dust from the atmosphere. In addition to objectives related to human exploration, the MECA data set will be rich in information relevant to basic geology, paleoclimate, and exobiology issues. To understand both contemporaneous and ancient processes on Mars, the mineralogy, petrology, and reactivity of Martian surface materials should be constrained. The MECA experiment will shed light on these quantities through its combination of chemistry and microscopy. MECA will be capable of measuring the composition of ancient surface water environments, observing microscopic evidence of geological (and biological?) processes, inferring soil and dust transport, comminution and weathering mechanisms, and characterizing soil horizons that might be encountered during excavation.

Hecht, M. H.↗

NASA NTRS1999

Soil Analysis Micro-Mission Concepts Derived from the MSP 2001 Mars Environmental Compatibility Assessment (MECA)

The Mars Environmental Compatibility Assessment (MECA) will evaluate the Martian environment for soil and dust-related hazards to human exploration as part of the Mars Surveyor Program 2001 Lander. The integrated MECA payload contains a wet-chemistry laboratory, a microscopy station, an electrometer to characterize the electrostatic environment, and arrays of material patches to study abrasion and adhesion. Heritage will be all-important for low cost micro-missions, and adaptations of instruments developed for the Pathfinder, '98 and '01 Landers should be strong contenders for '03 flights. This talk has three objectives: (1) Familiarize the audience with MECA instrument capabilities; (2) present concepts for stand-alone and/or mobile versions of MECA instruments; and (3) broaden the context of the MECA instruments from human exploration to a comprehensive scientific survey of Mars. Due to time limitations, emphasis will be on the chemistry and microscopy experiments. Ion-selective electrodes and related sensors in MECA's wet-chemistry laboratory will evaluate total dissolved solids, redox potential, pH, and the concentration of many soluble ions and gases in wet Martian soil. These electrodes can detect potentially dangerous heavy-metal ions, emitted pathogenic gases, and the soil's corrosive potential, and experiments will include cyclic voltammetry and anodic stripping. For experiments beyond 2001, enhancements could allow multiple use of the cells (for mobile experiments) and reagent addition (for quantitative mineralogical and exobiological analysis). MECA's microscopy station combines optical and atomic-force microscopy (AFM) in an actively focused, controlled illumination environment to image particles from millimeters to nanometers in size. Careful selection of substrates allows controlled experiments in adhesion, abrasion, hardness, aggregation, magnetic and other properties. Special tools allow primitive manipulation (brushing and scraping) of samples. Soil particle properties including size, shape, color, hardness, adhesive potential (electrostatic and magnetic), will be determined using an array of sample receptacles and collection substrates. The simple, rugged atomic-force microscope will image in the submicron size range and has the capability of performing a particle-by-particle analysis of the dust and soil. Future implementations might enhance the optical microscopy with spectroscopy, or incorporate advanced AFM techniques for thermogravimetric and chemical analysis.

Hecht, M. H.↗

NASA NTRS1999

Immune Function and Reactivation of Latent Viruses

A major concern associated with long-duration space flight is the possibility of infectious diseases posing an unacceptable medical risk to crew members. One major hypothesis addressed in this project is that space flight will cause alterations in the immune system that will allow latent viruses that are endogenous in the human population to reactivate and shed to higher levels than normal, which may affect the health of crew members. The second major hypothesis being examined is that the effects of space flight will alter the mucosal immune system, the first line of defense against many microbial infections, including herpesviruses, polyomaviruses, and gastroenteritis viruses, rendering crew members more susceptible to virus infections across the mucosa. We are focusing the virus studies on the human herpesviruses and polyomaviruses, important pathogens known to establish latent infections in most of the human population. Both primary infection and reactivation from latent infection with these groups of viruses (especially certain herpesviruses) can cause a variety of illnesses that result in morbidity and, occasionally, mortality. Both herpesviruses and polyomaviruses have been associated with human cancer, as well. Effective vaccines exist for only one of the eight known human herpesviruses and available antivirals are of limited use. Whereas normal individuals display minimal consequences from latent viral infections, events which alter immune function (such as immunosuppressive therapy following solid organ transplantation) are known to increase the risk of complications as a result of viral reactivations.

Butel, Janet S.↗

NASA NTRS1999

DNA Probe Design for Preflight and Inflight Microbial Monitoring

Crew health is a dominant issue in manned space flight. Microbiological concerns, in particular, have repeatedly emerged as determinants of flight readiness. For example, in at least one case, suspected contamination of the potable water supply nearly forced a launch delay. In another instance, a crew member's urinary tract infection nearly led to early termination of the mission, in part due to the difficulty of accurately diagnosing the nature of the infection in-flight. Microbial problems are an increasing concern with the trend towards longer-duration missions. It is essential to the success of such missions that systems that deliver acceptable quality of air and water during the anticipated lifetime of the spacecraft be available. As mission duration and resupply intervals increase, it will be necessary to rely on advanced life support systems which incorporate both biological and physical-chemical recycling methods for air and water as well as provide food for the crew. It therefore is necessary to develop real-time, robust, in-flight monitoring procedures that are sensitive enough to detect less than 100 CFU (colony forming units) of bacteria per 100 milliliters of water. It would be desirable if the monitoring system could be readily "reprogrammed" to identify specific pathogens if an in-flight incident were to occur. Thus, the monitoring technology must simultaneously detect many organisms of interest, be subject to miniaturization and be highly automated The long range goal of project is to develop such monitoring systems.

Fox, George E.↗

NASA NTRS1999

Starch Metabolism in Space-Grown Soybean Seedlings

The focus of this research was the study of sugar metabolism in soybean plants that had been in a clinorotation condition. The scope of activities was broadened greatly after the onset of the award. This broadening added two major research foci: 1) B-PAC: Photosynthetic activity of Brassica rapa; and 2) SOYPAT: The effects of microgravity on the interaction of a fungal root pathogen with soybean. Substantial investment and activity was also focused on the training of the astronaut team to conduct these experiments during orbital spaceflight.

Guikema, James A.↗

NASA NTRS2000

Options in Extraterrestrial Sample Handling and Study

This presentation mentions important service functions such as: sample preservation, hazard assessment, and handling. It also discuss how preliminary examination of samples is necessary for sample hazard assessment and for sample allocations. Clean facilities and clean sample handling are required. Conflicts, cross contamination issues will be present and need to be resolved. Extensive experience is available for extraterrestrial samples and must be sought and applied. Extensive experience is available in studies of pathogenicity and must be sought and applied as necessary. Advisory and oversight structures must also be in place

Papanastassiou, Dimitri A.↗

NASA NTRS1999

Bring'em Back Alive or at Least Carefully

Within the next decade, the world's space agencies plan to launch a variety of robotic spacecraft that will return samples from the surface of Mars, the tail of a comet, the nucleus of a comet, the surface of an asteroid, and the solar wind. Most of these places are not considered likely spots for life, but any mission returning from a location with the potential for harboring life will require special containment and handling because of the possible inclusion of living entities within returned samples. In its 1997 report on sample return from Mars, the Space Studies Board of the National Research Council (NRC) noted that the only risk of significant adverse effects would be from returning a replicating organism. Furthermore, the report noted: 'While the probability of returning a replicating biological entity in a sample from Mars' is judged to be low and the risk of pathogenic or ecological effects is lower still, the risk is not zero. Therefore, it is reasonable that NASA adopt a prudent approach, erring on the side of caution and safety when dealing with returned samples. More recently, a 1998 NRC report on small solar system bodies (asteroids, comets, planetary satellites, and interplanetary dust) recommended a similarly cautious approach for samples returned from anywhere else within the solar system that could have environmental conditions conducive for harboring life. We have not detected life elsewhere in the solar system, at least not yet. Nonetheless, the rationale behind the conservative approach to sample handling is similar to the environmental, health, and safety measures taken on Earth when transporting or handling infectious agents or importing non-native organisms to a new area. Better safe than sorry.

Race, Margaret S.↗

NASA NTRS2001

Remote Sensing, GIS, and Vector-Borne Disease

The concept of global climate change encompasses more than merely an alteration in temperature; it also includes spatial and temporal covariations in precipitation and humidity, and more frequent occurrence of extreme weather events. The impact of these variations, which can occur at a variety of temporal and spatial scales, could have a direct impact on disease transmission through their environmental consequences for pathogen, vector, and host survival, as well as indirectly through human demographic and behavioral responses. New and future sensor systems will allow scientists to investigate the relationships between climate change and environmental risk factors at multiple spatial, temporal and spectral scales. Higher spatial resolution will provide better opportunities for mapping urban features previously only possible with high resolution aerial photography. These opportunities include housing quality (e.g., Chagas'disease, leishmaniasis) and urban mosquito habitats (e.g., dengue fever, filariasis, LaCrosse encephalitis). There are or will be many new sensors that have higher spectral resolution, enabling scientists to acquire more information about parameters such as soil moisture, soil type, better vegetation discrimination, and ocean color, to name a few. Although soil moisture content is now detectable using Landsat, the new thermal, shortwave infrared, and radar sensors will be able to provide this information at a variety of scales not achievable using Landsat. Soil moisture could become a key component in transmission risk models for Lyme disease (tick survival), helminthiases (worm habitat), malaria (vector-breeding habitat), and schistosomiasis (snail habitat).

Beck, Louisa R.↗

NASA NTRS2001

Visualization of Microbial Biomarkers by Scanning Electron Microscopy

We are developing tools to link the biochemical structure of selected biomarkers with putative biogenic structures observed in mineralized samples. The detection of evidence of life on Mars and other planets will rely on methods that can discriminate compounds formed exclusively by living organisms. While biogenic compounds, such as amino acids and nucleotides have been discovered in extraterrestrial sources, such as meteorites and comets, their formation can be explained by abiotic means. The formation of cellular structures, or more elaborate organic molecules, such as complex lipids, proteins or nucleic acids, however, is strongly correlated to the presence of even the most primitive life processes. Recent evidence lends support to the hypothesis that life may have once existed on Mars. Carbonate globules and ppm concentrations of polycyclic aromatic hydrocarbons (PAHs) have been described in ALH84001, a meteorite originating from Mars ejecta captured by Earth over 13,000 years ago. The localized high concentration of PAHs that follow an increasing gradient from the intact fusion crust towards the interior corresponds to microgram quantities of hydrocarbon. Even though ALH84001 and other similar meteorites have withstood the forces capable of ejecting rock through Mars' escape velocity, upon entering Earth's atmosphere, their core temperatures are likely not to have been raised significantly, as evidenced by the survival of remanent magnetic signatures. Ideal biomarkers of ancient or modern biological life would include molecules that are (or were) pervasive and highly resistant to degradation. Also, requisite methods of detection should be simple, extremely sensitive and broadly inclusive (NASA SP-530). Lipopolysaccharide (LPS), peptidoglycan or pseudopeptidoglycan and beta-glucan are microbial cell wall components which together cover the entire microbial spectrum of eubacteria, archea and fungi. They are all remarkably resistant to thermal degradation. Fortunately, many antimicrobial defense systems of higher organisms require sensitive detection to combat microbial pathogens. We employ here the primitive immune system of the evolutionarily ancient horseshoe crab, Limulus polyphemus. This species relies on multi-enzyme signal amplification detection of cell wall molecules and they can be applied to the development of useful detectors of life. An extension of this work includes the visualization of microbial signatures by labeling LAL components with chromogenic or electron dense markers. The protein Limulus Anti-LPS Factor (LALF) has an extremely high affinity for LPS. By coupling LALF binding with colloidal gold labels we demonstrate a correlation of the structures visible by electron microscopy with biochemical evidence of microbial cell wall materials. Pure silica particles were mixed with cultures of E. coli (10(exp 6) cfu/mL). Samples were washed sequentially with buffered saline, LALF, antibody to LALF and finally colloidal gold-labeled Protein A. Negative controls were not exposed to E. coli but received identical treatment otherwise. Samples were coated with carbon and imaged on a JEOL JSM-840 scanning electron microscope with LaB6 source in the back scatter mode with the JEOL annular back scatter detector. 20 nm-scale black spots in this contrast-reversed image originate from electrons back-scattered by gold atoms. Negative controls did not give any signal. Future work will expand application of this technique to soil simulants and mineralized rock samples.

Wainwright, Norman R.↗

NASA NTRS2002

Satellites as Shared Resources for Caribbean Climate and Health Studies

Remotely-sensed data and observations are providing powerful new tools for addressing climate and environment-related human health problems through increased capabilities for monitoring, risk mapping, and surveillance of parameters useful to such problems as vector-borne and infectious diseases, air and water quality, harmful algal blooms, UV (ultraviolet) radiation, contaminant and pathogen transport in air and water, and thermal stress. Remote sensing, geographic information systems (GIS), global positioning systems (GPS), improved computational capabilities, and interdisciplinary research between the Earth and health science communities are being combined in rich collaborative efforts resulting in more rapid problem-solving, early warning, and prevention in global health issues. Collaborative efforts among scientists from health and Earth sciences together with local decision-makers are enabling increased understanding of the relationships between changes in temperature, rainfall, wind, soil moisture, solar radiation, vegetation, and the patterns of extreme weather events and the occurrence and patterns of diseases (especially, infectious and vector-borne diseases) and other health problems. This increased understanding through improved information and data sharing, in turn, empowers local health and environmental officials to better predict health problems, take preventive measure, and improve response actions. This paper summarizes the remote sensing systems most useful for climate, environment and health studies of the Caribbean region and provides several examples of interdisciplinary research projects in the Caribbean currently using remote sensing technologies. These summaries include the use of remote sensing of algal blooms, pollution transport, coral reef monitoring, vectorborne disease studies, and potential health effects of African dust on Trinidad and Barbados.

Maynard, Nancy G.↗

NASA NTRS2002

Deciphering the Role of Desert Dust in the Climate Puzzle: The Mediterranean Israeli Dust Experiment (MEIDEX)

Numerous studies have shown that aerosol particles may be one of the primary agents that can offset the climate warming induced by the increase in the amount of atmospheric greenhouse gases. Desert aerosols are probably the most abundant and massive type of aerosol particles that are present in the atmosphere worldwide. These aerosols are carried over large distances and have various global impacts. They interact with clouds, impact the efficiency of their rain production and change their optical properties. They constitute one of the primary sources of minerals for oceanic life and influence the health of coral reefs. They have direct effects on human health, especially by inducing breathing difficulties in children. It was lately discovered that desert particles carry pathogens from the Sahara desert over the Atlantic Ocean, a fact that may explain the migration of certain types of diseases. Aerosols not only absorb solar radiation but also scatter it, so that their climatic effect is influenced not only by their physical properties and height distribution but also by the reflectivity of the underlying surface. This latter property changes greatly over land and is low over ocean surfaces. Aerosol plumes are emitted from discrete, sporadic sources in the desert areas of the world and are transported worldwide by the atmosphere's wind systems. For example, Saharan dust reaches Mexico City, Florida, Ireland, Switzerland and the Mediterranean region, while Asian dust reaches Alaska, Hawaii and the continental United States. This means that in order to assess its global effects, one must observe dust from space. The Space Shuttle is a unique platform, because it flies over the major deserts of our planet, enabling measurements and remote sensing of the aerosols as they travel from source to sink regions. Such efforts must always be accompanied by in-situ data for validation and calibration, with direct sampling of the airborne particles. MEIDEX is a joint project of the Israel Space Agency (ISA) and NASA, under a cooperation agreement between the two agencies.

Levin, Zev↗

NASA NTRS2002

In-Vessel Composting of Simulated Long-Term Missions Space-Related Solid Wastes

Reduction and stabilization of solid wastes generated during space missions is a major concern for the Advanced Life Support - Resource Recovery program at the NASA, Kennedy Space Center. Solid wastes provide substrates for pathogen proliferation, produce strong odor, and increase storage requirements during space missions. A five periods experiment was conducted to evaluate the Space Operation Bioconverter (SOB), an in vessel composting system, as a biological processing technology to reduce and stabilize simulated long-term missions space related solid-wastes (SRSW). For all periods, SRSW were sorted into components with fast (FBD) and slow (SBD) biodegradability. Uneaten food and plastic were used as a major FBD and SBD components, respectively. Compost temperature (C), CO2 production (%), mass reduction (%), and final pH were utilized as criteria to determine compost quality. In period 1, SOB was loaded with a 55% FBD: 45% SBD mixture and was allowed to compost for 7 days. An eleven day second composting period was conducted loading the SOB with 45% pre-composted SRSW and 55% FBD. Period 3 and 4 evaluated the use of styrofoam as a bulking agent and the substitution of regular by degradable plastic on the composting characteristics of SRSW, respectively. The use of ceramic as a bulking agent and the relationship between initial FBD mass and heat production was investigated in period 5. Composting SRSW resulted in an acidic fermentation with a minor increase in compost temperature, low CO2 production, and slightly mass reduction. Addition of styrofoam as a bulking agent and substitution of regular by biodegradable plastic improved the composting characteristics of SRSW, as evidenced by higher pH, CO2 production, compost temperature and mass reduction. Ceramic as a bulking agent and increase the initial FBD mass (4.4 kg) did not improve the composting process. In summary, the SOB is a potential biological technology for reduction and stabilization of mission space-related solid wastes. However, the success of the composting process may depend of the physical characteristics (particle size, porosity, structure, texture) of the SBD components which would require pre-processing of solid wastes before placing them in the SOB.

Rodriguez-Carias, Abner A.↗