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At least 145 records · Page 8

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NASA NTRS1989

Peroxides and the survivability of microorganisms on the surface of Mars

This paper discusses the possibility that any terrestrial microorganisms brought to Mars might survive the unhospitable environment of that planet, with special attention given to the effects of highly oxidizing material that is now known to cover the Martian surface. Data obtained by the gas exchange experiment on Viking indicate that, if all of the released oxygen is assumed to come from H2O2, the concentrations of H2O2 on Mars range from 25 to 250 ppm. Laboratory data indicate that certain soil bacteria are able to survive and grow to stationary phase at H2O2 concentrations as high as 30,000, indicating that, if there is H2O2 at the level of 250 ppm or even an order of magnitude greater on the Martian surface, this fact alone would not make the surface of Mars self-sterilizing.

Mancinelli, Rocco L.↗

NASA NTRS1992

Autonomous support for microorganism research in space

A preliminary design for performing on orbit, autonomous research on microorganisms and cultured cells/tissues is presented. An understanding of gravity and its effects on cells is crucial for space exploration as well as for terrestrial applications. The payload is designed to be compatible with the Commercial Experiment Transporter (COMET) launch vehicle, an orbiter middeck locker interface, and with Space Station Freedom. Uplink/downlink capabilities and sample return through controlled reentry are available for all carriers. Autonomous testing activities are preprogrammed with in-flight reprogrammability. Sensors for monitoring temperature, pH, light, gravity levels, vibrations, and radiation are provided for environmental regulation and experimental data collection. Additional experimental data acquisition includes optical density measurement, microscopy, video, and film photography. On-board full data storage capabilities are provided. A fluid transfer mechanism is utilized for inoculation, sampling, and nutrient replenishment of experiment cultures. In addition to payload design, representative experiments were developed to ensure scientific objectives remained compatible with hardware capabilities. The project is defined to provide biological data pertinent to extended duration crewed space flight including crew health issues and development of a Controlled Ecological Life Support System (CELSS). In addition, opportunities are opened for investigations leading to commercial applications of space, such as pharmaceutical development, modeling of terrestrial diseases, and material processing.

Fleet, Mary L.↗

NASA NTRS1992

Autonomous support for microorganism research in space

A preliminary design for performing on-orbit, autonomous research on microorganisms and cultured cells/tissues is presented. An understanding of gravity and its effects on cells is crucial for space exploration as well as for terrestrial applications. The payload is designed to be compatible with the COMmercial Experiment Transported (COMET) launch vehicle, an orbiter middeck locker interface, and with Space Station Freedom. Uplink/downlink capabilities and sample return through controlled reentry are available for all carriers. Autonomous testing activities are preprogrammed with inflight reprogrammability. Sensors for monitoring temperature, pH, light, gravity levels, vibration, and radiation are provided for environmental regulation and experimental data collection. Additional experiment data acquisition includes optical density measurement, microscopy, video, and file photography. Onboard full data storage capabilities are provided. A fluid transfer mechanism is utilized for inoculation, sampling, and nutrient replenishment of experiment cultures. In addition to payload design, representative experiments were developed to ensure scientific objectives remained compatible with hardware capabilities. The project is defined to provide biological data pertinent to extended duration crewed space flight including crew health issues and development of a Controlled Ecological Life Support System (CELSS). In addition, opportunities are opened for investigations leading to commercial applications of space, such as pharmaceutical development, modeling of terrestrial diseases, and material processing.

Luttges, M. W.↗

NASA NTRS1994

Dynamics of microorganism populations in recirculating nutrient solutions

This overview covers the basic microbial ecology of recirculating hydroponic solutions. Examples from NASA and Soviet Controlled Ecological Life Support Systems (CELSS) tests and the commercial hydroponic industry will be used. The sources of microorganisms in nutrient solutions include air, water, seeds, plant containers and plumbing, biological vectors, and personnel. Microbial fates include growth, death, and emigration. Important microbial habitats within nutrient delivery systems are root surfaces, hardware surfaces (biofilms), and solution suspension. Numbers of bacteria on root surfaces usually exceed those from the other habitats by several orders of magnitude. Gram negative bacteria dominate the microflora with fungal counts usually much lower. Trends typically show a decrease in counts with increasing time unless stressed plants increase root exudates. Important microbial activities include carbon mineralization and nitrogen transformations. Important detrimental interactions include competition with plants, and human and plant pathogenesis.

Strayer, R. F.↗

NASA NTRS1994

Dynamics of microorganism populations in recirculating nutrient solutions

This overview covers the basic microbial ecology of recirculating hydroponic solutions. Examples from NASA and Soviet CELSS tests and the commercial hydroponic industry will be used. The sources of microorganisms in nutrient solutions include air, water, seeds, plant containers and plumbing, biological vectors, and personnel. Microbial fates include growth, death, and emigration. Important microbial habitats within nutrient delivery systems are root surfaces, hardware surfaces (biofilms), and solution suspension. Numbers of bacteria on root surfaces usually exceed those from the other habitats by several orders of magnitude. Gram negative bacteria dominate the microflora with fungal counts usually much lower. Trends typically show a decrease in counts with increasing time unless stressed plants increase root exudates. Important microbial activities include carbon mineralization and nitrogen transformations. Important detrimental interactions include competition with plants, and human and plant pathogenesis.

Strayer, R. F.↗

NASA NTRS1999

Microorganisms from Permafrost Viable and Detectable by 16SRNA Analysis: A Model for Mars

Preliminary studies of Arctic and Antarctic permafrost have shown that this environment harbors microorganisms which can be isolated in pure culture, and that these organisms can survive for a long period of time (up to 20 Ma) in permafrost. It is believed that the permanent subzero temperatures in permafrost and ice environments are the main parameters ensuring the longevity of microbes. In this project we studied permafrost cores from different areas of the Siberian Arctic and Antarctic, with ages from several thousand years up to several millions years (Ma). In general, Antarctic permafrost has a higher sand content, while Siberian permafrost has a texture more characteristic of clay or normal soil. Additional information is contained in the original extended abstract.

Tsapin, A. I.↗

NASA NTRS1997

Density and composition of microorganisms during long-term (418 day) growth of potato using biologically reclaimed nutrients from inedible plant biomass

A study evaluating alternative methods for long term operation of biomass production systems was recently completed at the Kennedy Space Center (KSC). The 418-day study evaluated repeated batch versus mixed-aged production of potato grown on either standard 1/2-strength Hoagland's nutrient solution or solutions including nutrients recycled from inedible plant material. The long term effects of closure and recycling on microbial dynamics were evaluated by monitoring the microbial communities associated with various habitats within the plant growth system (i.e., plant roots, nutrient solution, biofilms within the hydroponic systems, atmosphere, and atmospheric condensate). Plate count methods were used to enumerate and characterize microorganisms. Microscopic staining methods were used to estunate total cell densities. The primary finding was that the density and composition of microbial communities associated with controlled environmental plant growth systems are stable during long term operation. Continuous production resulted in slightly greater stability. Nutrient recycling, despite the addition of soluble organic material from the waste processing system, did not significantly increase microbial density in any of the habitats.

NASA Center KSC↗

NASA NTRS1987

The evolution of glutathione metabolism in phototrophic microorganisms

Of the many roles ascribed to glutathione (GSH) the one most clearly established is its role in the protection of higher eucaryotes against oxygen toxicity through destruction of thiol-reactive oxygen byproducts. If this is the primary function of GSH then GSH metabolism should have evolved during or after the evolution of oxygenic photosynthesis. That many bacteria do not produce GSH is consistent with this view. In the present study we have examined the low-molecular-weight thiol composition of a variety of phototrophic microorganisms to ascertain how evolution of GSH production is related to evolution of oxygenic photosynthesis. Cells were extracted in the presence of monobromobimane (mBBr) to convert thiols to fluorescent derivatives, which were analyzed by high-pressure liquid chromatography. Significant levels of GSH were not found in the green bacteria (Chlorobium thiosulfatophilum and Chloroflexus aurantiacus). Substantial levels of GSH were present in the purple bacteria (Chromatium vinosum, Rhodospirillum rubrum, Rhodobacter sphaeroides, and Rhodocyclus gelatinosa), the cyanobacteria [Anacystis nidulans, Microcoleus chthonoplastes S.G., Nostoc muscorum, Oscillatoria amphigranulata, Oscillatoria limnetica, Oscillatoria sp. (Stinky Spring, Utah), Oscillatoria terebriformis, Plectonema boryanum, and Synechococcus lividus], and eucaryotic algae (Chlorella pyrenoidsa, Chlorella vulgaris, Euglena gracilis, Scenedesmus obliquus, and Chlamydomonas reinhardtii). Other thiols measured included cysteine, gamma-glutamylcysteine, thiosulfate, coenzyme A, and sulfide; several unidentified thiols were also detected. Many of the organisms examined also exhibited a marked ability to reduce mBBr to syn-(methyl,methyl)bimane, an ability that was quenched by treatment with 2-pyridyl disulfide or 5,5'-bisdithio-(2-nitrobenzoic acid) prior to reaction with mBBr. These observations indicate the presence of a reducing system capable of electron transfer to mBBr and reduction of reactive disulfides. The distribution of GSH in phototrophic eubacteria indicates that GSH synthesis evolved at or around the time that oxygenic photosynthesis evolved.

NASA Discipline Exobiology↗

NASA NTRS1988

Temperature response of Antarctic cryptoendolithic photosynthetic microorganisms

Growth responses to temperatures between 12.5 [degrees] C and 25 degrees C were determined for five photosynthetic microorganisms isolated from the Ross Desert cryptoendolithic community. Among eukaryotic algae, two strains of Trebouxia sp. have an upper temperature limit of 20 degrees C, and two strains of Hemichloris antarctica of 25 degrees C. The cyanobacterium Chroococcidiopsis sp., in contrast, grows at temperatures above 25 degrees C. These and earlier studies suggest that the eukaryotic algae of the Antarctic cryptoendolithic community have an upper temperature limit near 25 degrees C.

NASA Discipline Exobiology↗

NASA NTRS2004

Phylogenetic and physiological diversity of microorganisms isolated from a deep greenland glacier ice core

We studied a sample from the GISP 2 (Greenland Ice Sheet Project) ice core to determine the diversity and survival of microorganisms trapped in the ice at least 120,000 years ago. Previously, we examined the phylogenetic relationships among 16S ribosomal DNA (rDNA) sequences in a clone library obtained by PCR amplification from genomic DNA extracted from anaerobic enrichments. Here we report the isolation of nearly 800 aerobic organisms that were grouped by morphology and amplified rDNA restriction analysis patterns to select isolates for further study. The phylogenetic analyses of 56 representative rDNA sequences showed that the isolates belonged to four major phylogenetic groups: the high-G+C gram-positives, low-G+C gram-positives, Proteobacteria, and the Cytophaga-Flavobacterium-Bacteroides group. The most abundant and diverse isolates were within the high-G+C gram-positive cluster that had not been represented in the clone library. The Jukes-Cantor evolutionary distance matrix results suggested that at least 7 isolates represent new species within characterized genera and that 49 are different strains of known species. The isolates were further categorized based on the isolation conditions, temperature range for growth, enzyme activity, antibiotic resistance, presence of plasmids, and strain-specific genomic variations. A significant observation with implications for the development of novel and more effective cultivation methods was that preliminary incubation in anaerobic and aerobic liquid prior to plating on agar media greatly increased the recovery of CFU from the ice core sample.

Bacteria/classification/genetics/isolation & purif↗

NASA NTRS1999

Syntrophus aciditrophicus sp. nov., a new anaerobic bacterium that degrades fatty acids and benzoate in syntrophic association with hydrogen-using microorganisms

Strain SBT is a new, strictly anaerobic, gram-negative, nonmotile, non-sporeforming, rod-shaped bacterium that degrades benzoate and certain fatty acids in syntrophic association with hydrogen/formate-using microorganisms. Strain SBT produced approximately 3 mol of acetate and 0.6 mol of methane per mol of benzoate in coculture with Methanospirillum hungatei strain JF1. Saturated fatty acids, some unsaturated fatty acids, and methyl esters of butyrate and hexanoate also supported growth of strain SBT in coculture with Desulfovibrio strain G11. Strain SBT grew in pure culture with crotonate, producing acetate, butyrate, caproate, and hydrogen. The molar growth yield was 17 +/- 1 g cell dry mass per mol of crotonate. Strain SBT did not grow with fumarate, iron(III), polysulfide, or oxyanions of sulfur or nitrogen as electron acceptors with benzoate as the electron donor. The DNA base composition of strain SBT was 43.1 mol% G+C. Analysis of the 16 S rRNA gene sequence placed strain SBT in the delta-subdivision of the Proteobacteria, with sulfate-reducing bacteria. Strain SBT was most closely related to members of the genus Syntrophus. The clear phenotypic and genotypic differences between strain SBT and the two described species in the genus Syntrophus justify the formation of a new species, Syntrophus aciditrophicus.

Non-NASA Center↗

NASA NTRS1996

Diversity of eukaryotic microorganisms: computer-based resources, "The Handbook of Protoctista" and its "Glossary"

The kingdom Protoctista comprises some 30 phyla, including the eukaryotic anaerobes that permanently lack mitochondria, the Phylum Archaeprotista, with its three classes: (i) Archamoebae, e.g., Pelomyxa, Mastigina, (ii) Metamonada, e.g., Giardia, Pyrsonympha, and (iii) Parabasalia, e.g., Trichomonas, Calonympha, and the Phylum Microspora (Microsporidia), e.g., Vairimorpha. These and all algae, protozoa, labyrinthulids, "water molds" (oomycota, plasmodiophorans, hyphochytrids, chytrids, etc.) and other eukaryotes excluded from plants, animals and fungi are detailed in the Handbook of Protoctista. The Illustrated Glossary of Protoctista contains descriptions of the morphology and taxonomy of these microorganisms, including the many equivalent and homologous structures with different names. The Glossary has also been made into a Macintosh-compatible CD-ROM disk.

NASA Discipline Number 52-30↗

NASA NTRS1995

A novel approach for monitoring genetically engineered microorganisms by using artificial, stable RNAs

Further improvements in technology for efficient monitoring of genetically engineered microorganisms (GEMs) in the environment are needed. Technology for monitoring rRNA is well established but has not generally been applicable to GEMs because of the lack of unique rRNA target sequences. In the work described herein, it is demonstrated that a deletion mutant of a plasmid-borne Vibrio proteolyticus 5S rRNA gene continues to accumulate to high levels in Escherichia coli although it is no longer incorporated into 70S ribosomes. This deletion construct was subsequently modified by mutagenesis to create a unique recognition site for the restriction endonuclease BstEII, into which new sequences could be readily inserted. Finally, a novel 17-nucleotide identifier sequence from Pennisetum purpureum was embedded into the construct to create an RNA identification cassette. The artificial identifier RNA, expressed from this cassette in vivo, accumulated in E. coli to levels comparable to those of wild-type 5S rRNA without being seriously detrimental to cell survival in laboratory experiments and without entering the ribosomes. These results demonstrate that artificial, stable RNAs containing sequence segments remarkably different from those present in any known rRNA can be designed and that neither the deleted sequence segment nor ribosome incorporation is essential for accumulation of an RNA product.

Non-NASA Center↗

NASA NTRS2005

Fossil Microorganisms in Archaean deposits of Northern Karelia

Newly found biomorphic microstructures from the Upper Archaean (lopian) rocks from Northern Karelia are described. The presence of various microorganisms of bacterial nature and even cyanobacteria (and possibly eukaryotic forms) is suggested. The necessity of employing methods of electron microscopy, as well as traditional methods, while studying the very early manifestations of life in Archaean and Early Proterozoic is noted.

Astafieva, M. M.↗

NASA NTRS2006

Fossils of Prokaryotic Microorganisms in the Orgueil Meteorite

The Orgueil CII meteorite, which fell in southern France on the evening of May 14, 1864, has been one of the most extensively studied of all known carbonaceous meteorites. Field Emission Scanning Electron Microscopy (FESEM) studies of freshly fractured interior surfaces of the Orgueil meteorite have resulted in the detection of the fossilized remains of a large and diverse population of filamentous prokaryotic microorganisms. The taphonomy and the diverse modes of the preservation of these remains ,are diverse. Some of the remains exhibit carbonization of a hollow sheath and in other cases the remains are permineralized with water-soluble evaporite minerals, such as magnesium sulfate or ammonium salts. After the sample is fractured and the interior surfaces are exposed to the atmospheric moisture, some of these friable remains have been observed to exhibit significant alterations in appearance with time. Images are presented to document the changes that have been observed in some forms within the past two years. Images and EDS spectral data will also be presented to document the studies carried out on abiotic forms to search for possible nonbiological interpretations of the indigenous filamentous microstructures that have been found in the Orgueil meteorite. Images and EDS data will be presented showing the size, size range, morphology and chemical compositions of abiotic microstructures found in native crystalline and fibrous Epsomites from Poison Lake, Washington, USA and Catalayud, Zaragoza, Aragon, Spain. Many of these embedded forms are consistent in size and microstructure with cyanobacteria morphotypes. Some of the forms are exhibit known characteristics differentiation of cells, and reproductive structures of filamentous trichomic prokaryotes (bacteria and cyanobacteria) and the degraded remains of microfibrils associated with sheaths of cyanobacteria. In this paper, recently obtained comparative images and EDS data will be presented for the mineralized remains found in the Orgueil meteorite and these forms will be compared with known cyanobacteria of the family Oscillatoriaceae. Many of the indigenous filamentous microstructures found in Orgueil filaments are dramatically different from the abiotic microstructures found in the fibrous epsomites and these forms are interpreted as the molds, casts and the well preserved permineralized remains (microfossils) of filamentous prokaryotes that are embedded in and indigenous to the matrix of the Orgueil CI1 carbonaceous meteorite.

Hoover, Richard B.↗

NASA NTRS2007

Super-long Anabiosis of Ancient Microorganisms in Ice and Terrestrial Models for Development of Methods to Search for Life on Mars, Europa and other Planetary Bodies

Successful missions to Mars, Europe and other bodies of the Solar system have created a prerequisite to search for extraterrestrial life. The first attempts of microbial life detection on the Martian surface by the Viking landed missions gave no biological results. Microbiological investigations of the Martian subsurface ground ice layers seem to be more promising. It is well substantiated to consider the Antarctic ice sheet and the Antarctic and Arctic permafrost as terrestrial analogues of Martian habitats. The results of our long-standing microbiological studies of the Antarctic ice would provide the basis for detection of viable microbial cells on Mars. Our microbiological investigations of the deepest and thus most ancient strata of the Antarctic ice sheet for the first time gave evidence for the natural phenomenon of long-term anabiosis (preservation of viability and vitality for millennia years). A combination of classical microbiological methods, epifluorescence microscopy, SEM, TEM, molecular diagnostics, radioisotope labeling and other techniques made it possible for us to obtain convincing proof of the presence of pro- and eukaryotes in the Antarctic ice sheet. In this communication, we will review and discuss some critical issues related to the detection of viable microorganisms in cold terrestrial environments with regard to future searches for microbial life and/or its biological signatures on extraterrestrial objects.

Abyzov, S. S.↗

NASA NTRS2012

Methods for determining the genetic affinity of microorganisms and viruses

Selecting which sub-sequences in a database of nucleic acid such as 16S rRNA are highly characteristic of particular groupings of bacteria, microorganisms, fungi, etc. on a substantially phylogenetic tree. Also applicable to viruses comprising viral genomic RNA or DNA. A catalogue of highly characteristic sequences identified by this method is assembled to establish the genetic identity of an unknown organism. The characteristic sequences are used to design nucleic acid hybridization probes that include the characteristic sequence or its complement, or are derived from one or more characteristic sequences. A plurality of these characteristic sequences is used in hybridization to determine the phylogenetic tree position of the organism(s) in a sample. Those target organisms represented in the original sequence database and sufficient characteristic sequences can identify to the species or subspecies level. Oligonucleotide arrays of many probes are especially preferred. A hybridization signal can comprise fluorescence, chemiluminescence, or isotopic labeling, etc.; or sequences in a sample can be detected by direct means, e.g. mass spectrometry. The method's characteristic sequences can also be used to design specific PCR primers. The method uniquely identifies the phylogenetic affinity of an unknown organism without requiring prior knowledge of what is present in the sample. Even if the organism has not been previously encountered, the method still provides useful information about which phylogenetic tree bifurcation nodes encompass the organism.

Fox, George E.↗

NASA NTRS2014

Microorganisms in the Stratosphere (MIST): In-flight Sterilization with UVC Leds

The stratosphere (10 km to 50 km above sea level) is a unique place on Earth for astrobiological studies of microbes in extreme environments due to the combination of harsh conditions (high ultraviolet radiation, low pressure, desiccation, and low temperatures). Microorganisms in the Stratosphere (MIST) will attempt to characterize the diversity of microbes at these altitudes using a balloon collection device on a meteorological weather balloon. A major challenge of such an aerobiology study is the potential for ground contamination that makes it difficult to distinguish between collected microbes and contaminants. One solution is to use germicidal ultraviolet light emitting diodes (UV LEDs) to sterilize the collection strip. To use this solution, an optimal spatial arrangement of the lights had to be determined to ensure the greatest chance of complete sterilization within the 30 to 60 minute time of balloon ascent. A novel, 3D-printed test stand was developed to experimentally determine viable Bacillus pumilus SAFR-032 spore reduction after exposure to ultraviolet radiation at various times, angles, and distances. Taken together, the experimental simulations suggested that the UV LEDs on the MIST flight hardware should be active for at least 15 minutes and mounted within 4 cm of the illuminated surface at any angle to achieve optimal sterilization. These findings will aid in the production of the balloon collection device to ensure pristine stratospheric microbial samples are collected. Flight hardware capable of in-flight self-sterilization will enable future life detection missions to minimize both forward contamination and false positives.

Exobiology↗