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At least 145 records · Page 8

Preliminary Characterization of a Microbial Community of Rock Varnish from Death Valley, California

Rock varnish (also referred to as desert varnish in the literature because it is particularly noticeable in desert environments) is a dark, thin (typically 50-500 m thick), layered veneer composed of clay minerals cemented together by oxides and hydroxides of manganese and iron. Some scientists suggest that varnish may provide a historical record of environmental processes such as global warming and long-term climate change. However, despite more than 30 years of study using modern microanalytical and microbial culturing techniques, the nucleation and growth mechanisms of rock varnish remain a mystery. Rock varnish is of interest to the Mars science community because a varnish-like sheen has been reported on the rocks at the Viking Lander sites. It therefore important for us to understand the formation mechanisms of terrestrial varnish abiotic, biotic, or a combination of the two -- as this understanding may give us clues concerning the chemical and physical processes occurring on the surface of Mars. It is strongly believed by some in the biogeochemistry community that microbes have a role in forming rock varnish, and iron- and manganese-oxidation by microbes isolated from varnish has been extensively investigated. Only two of these studies have investigated the microbial genetics of varnish. These studies examined the morphological, physiological and molecular characteristics of microbes that had previously been cultured from various rock varnishes and identified the cultivars using 16S rDNA sequencing techniques. However, it is well known that most of organisms existing in nature are refractory to cultivation, so many important organisms would have been missed. The currently described work investigates the genetics of rock varnish microbial community from a site in the Whipple Mtns., south of Death Valley, CA, near Parker, Arizona. We employed both cultural and molecular techniques to characterize the microorganisms found within the varnish and surrounding soil with the objectives of (a) identifying microorganisms potentially involved in varnish formation, and (b) discovering microorganisms that simply use the varnish as an extreme habitat.

Kuhlman, K. R.↗

Ecology and life history of an amoebomastigote, Paratetramitus jugosus, from a microbial mat: new evidence for multiple fission

Five microbial habitats (gypsum crust, gypsum photosynthetic community, Microcoleus mat, Thiocapsa scum, and black mud) were sampled for the presence of the euryhaline, rapidly growing amoebomastigote, Paratetramitus jugosus. Field investigations of microbial mats from Baja California Norte, Mexico, and Salina Bido near Matanzas, Cuba, reveal that P. jugosus is most frequently found in the Thiocapsa layer of microbial mats. Various stages of the life history were studied using phase-contrast, differential-interference, and transmission electron microscopy. Mastigote stages were induced and studied by electron microscopy; mastigotes that actively feed on bacteria bear two or more undulipodia. A three-dimensional drawing of the kinetid ("basal apparatus") based on electron micrographs is presented. Although promitoses were occasionally observed, it is unlikely that they can account for the rapid growth of P. jugosus populations on culture media. Dense, refractile, spherical, and irregular-shaped bodies were seen at all times in all cultures along with small mononucleate (approximately 2-7 micrometers diameter) amoebae. Cytochemical studies employing two different fluorescent stains for DNA (DAPI, mithramycin) verified the presence of DNA in these small bodies. Chromatin-like material seen in electron micrographs within the cytoplasm and blebbing off nuclei were interpreted to the chromatin bodies. Our interpretation, consistent with the data but not proven, is that propagation by multiple fission of released chromatin bodies that become small amoebae may occur in Paratetramitus jugosus. These observations are consistent with descriptions of amoeba propagules in the early literature (Hogue, 1914).

Non-NASA Center↗

Fourier transform-infrared spectroscopic methods for microbial ecology: analysis of bacteria, bacteria-polymer mixtures and biofilms

Fourier transform-infrared (FT-IR) spectroscopy has been used to rapidly and nondestructively analyze bacteria, bacteria-polymer mixtures, digester samples and microbial biofilms. Diffuse reflectance FT-IR (DRIFT) analysis of freeze-dried, powdered samples offered a means of obtaining structural information. The bacteria examined were divided into two groups. The first group was characterized by a dominant amide I band and the second group of organisms displayed an additional strong carbonyl stretch at approximately 1740 cm-1. The differences illustrated by the subtraction spectra obtained for microbes of the two groups suggest that FT-IR spectroscopy can be utilized to recognize differences in microbial community structure. Calculation of specific band ratios has enabled the composition of bacteria and extracellular or intracellular storage product polymer mixtures to be determined for bacteria-gum arabic (amide I/carbohydrate C-O approximately 1150 cm-1) and bacteria-poly-beta-hydroxybutyrate (amide I/carbonyl approximately 1740 cm-1). The key band ratios correlate with the compositions of the material and provide useful information for the application of FT-IR spectroscopy to environmental biofilm samples and for distinguishing bacteria grown under differing nutrient conditions. DRIFT spectra have been obtained for biofilms produced by Vibrio natriegens on stainless steel disks. Between 48 and 144 h, an increase in bands at approximately 1440 and 1090 cm-1 was seen in FT-IR spectra of the V. natriegens biofilm. DRIFT spectra of mixed culture effluents of anaerobic digesters show differences induced by shifts in input feedstocks. The use of flow-through attenuated total reflectance has permitted in situ real-time changes in biofilm formation to be monitored and provides a powerful tool for understanding the interactions within adherent microbial consortia.

NASA Discipline Number 61-10↗

Microbial mats, stromatolites and the rise of oxygen in the Precambrian atmosphere

Microbial mats are stratified communities that develop within the environmental microgradients established at the interfaces of water and solid substrates (Cohen, 1989). Stromatolites, the lithified remains of layered accumulations of microbial mats, occur in rocks as old as 3.5 Ga (Lowe, 1980; Walter et al., 1980). These lithified microbial communities represent the most ancient, widespread ecosystems known, and it is useful to explore their role in the accumulation of free oxygen in the ancient atmosphere.

NASA Program Exobiology↗

Cosmopolitan distribution of the large composite microbial mat spirochete, Spirosymplokos deltaeiberi

Inocula from organic-rich black muds immediately underlying intertidal laminated microbial mats dominated by Microcoleus chthonoplastes yielded large, variable diameter spirochetes. These unusual spirochetes, previously reported only from the Alfacs Peninsula at the delta of the Ebro river in northeast Spain, contain striking arrays of cytoplasmic granules packed into their protoplasmic cylinders. On several occasions, both in summer and winter, the huge spirochetes were recognized in samples from mats growing in the Sippewissett salt marsh at Woods Hole Massachusetts. They were also seen in similar samples from microbial mats at North Pond, Laguna Figueroa, Baja California Norte, Mexico. The identity of these spirochetes was confirmed by electron microscopy: number and disposition of flagella, composite structure, measurements of their distinctive cytoplasmic granules. The granules, larger, more conspicuous and present in addition to ribosomes, are hypothesized to contain ATPases. As culture conditions worsen, these spirochetes retract into membrane-bounded round bodies in which they form refractile inclusions. From morphology and behavior we conclude the North American spirochetes from both Atlantic and Pacific intertidal microbial mats are indistinguishable from those at the delta of the Ebro river. We conclude a cosmopolitan distribution for Spirosymplokos deltaeiberi.

NASA Discipline Exobiology↗

A model for diurnal patterns of carbon fixation in a Precambrian microbial mat based on a modern analog

Microbial mat communities are one of the first and most prevalent biological communities known from the Precambrian fossil record. These fossil mat communities are found as laminated sedimentary rock structures called stromatolites. Using a modern microbial mat as an analog for Precambrian stromatolites, a study of carbon fixation during a diurnal cycle under ambient conditions was undertaken. The rate of carbon fixation depends primarily on the availability of light (consistent with photosynthetic carbon fixation) and inorganic carbon, and not nitrogen or phosphorus. Atmospheric PCO2 is thought to have decreased from 10 bars at 4 Ga (10(9) years before present) to approximately 10(-4) bars today, implying a change in the availability of inorganic carbon for carbon fixation. Experimental manipulation of levels of inorganic carbon to levels that may have been available to Precambrian mat communities resulted in increased levels of carbon fixation during daylight hours. Combining these data with models of daylength during the Precambrian, models are derived for diurnal patterns of photosynthetic carbon fixation in a Precambrian microbial mat community. The models suggest that, even in the face of shorter daylengths during the Precambrian, total daily carbon fixation has been declining over geological time, with most of the decrease having occurred during the Precambrian.

Review↗

Characterization of microbial and chemical composition of shuttle wet waste with permanent gas and volatile organic compound analyses

Solid-waste treatment in space for Advanced Life Support, ALS, applications requires that the material can be safely processed and stored in a confined environment. Many solid-wastes are not stable because they are wet (40-90% moisture) and contain levels of soluble organic compounds that can contribute to the growth of undesirable microorganisms with concomitant production of noxious odors. In the absence of integrated Advanced Life Support systems on orbit, permanent gas, trace volatile organic and microbiological analyses were performed on crew refuse returned from the volume F "wet" trash of three consecutive Shuttle missions (STS-105, 109, and 110). These analyses were designed to characterize the short-term biological stability of the material and assess potential crew risks resulting from microbial decay processes during storage. Waste samples were collected post-orbiter landing and sorted into packaging material, food waste, toilet waste, and bulk liquid fractions deposited during flight in the volume F container. Aerobic and anaerobic microbial loads were determined in each fraction by cultivation on R2A and by acridine orange direct count (AODC). Dry and ash weights were performed to determine both water and organic content of the materials. Experiments to determine the aerobic and anaerobic biostability of refuse stored for varying periods of time were performed by on-line monitoring of CO2 and laboratory analysis for production of hydrogen sulfide and methane. Volatile organic compounds and permanent gases were analyzed using EPA Method TO15 by USEPA et al. [EPA Method TO15, The Determination of Volatile Organic Compounds (VOCs) in Ambient Air using SUMMA, Passivated Canister Sampling and Gas Chromatographic Analysis,1999] with gas chromatography/mass spectrometry and by gas chromatography with selective detectors. These baseline measures of waste stream content, labile organics, and microbial load in the volume F Shuttle trash provide data for waste subsystem analysis and atmospheric management within the ALS Project. Published by Elsevier Ltd on behalf of COSPAR.

NASA Center KSC↗

Anaerobic decomposition of cellulose by allcaliphilic microbial Community of Owens Lake, California

The study of communities of microbial extremophiles from anaerobic sediments of Owens Lake and Mono Lake in California has established the presence of active microbial cellulolytic processes in both lakes. The prior study of the microbial diversity in Mono Lake showed that the trophic chain of organic decomposition includes secondary anaerobes that were found to be previously unknown species (Spirochaeta americana, Tindallia californiensis, and Desulfonatronum thiodismutans). And as we published earlier, the secondary anaerobes of Owens Lakes were morphologically very similar to those of Mono Lake. However, continuing comparison of the physiology and genetics has led to the conclusion that some links of organic decomposition in the trophic chain of Owens Lake are represented by different unknown species. A new isolate of a sugarlytic spirochete from Owens Lake, which was morphologically very similar to S. americana isolated from Mono Lake, was found to have different metabolic capacity such as the lack of capability to produce hydrogen during glucose fermentation. Furthermore, from the same bacterial community (from Owens Lake) another sugarlytic spore-forming alkaliphile (strain SCA) was isolated in pure culture. Here we discuss the geology and chemistry of Owens Lake as a unique ecosystem of Astrobiological significance. This paper also presents some of the characteristics for the novel isolates and describes their participation in the process of cellulose degradation.

Pikuta, Elena V.↗

Microbial Characterization and Comparison of Isolates During the Mir and ISS Missions

Spacecraft represent a semi-closed ecosystem that provides a unique model of microbial interaction with other microbes, potential hosts, and their environment. Environmental samples from the Mir Space Station (1995-1998) and the International Space Station (ISS) (2000-Present) were collected and processed to provide insight into the characterization of microbial diversity aboard spacecraft over time and assess any potential health risks to the crew. All microbiota were isolated using standard media-based methodologies. Isolates from Mir and ISS were processed using various methods of analysis, including VITEK biochemical analysis, 16s ribosomal identification, and fingerprinting using rep-PCR analysis. Over the first 41 months of habitation, the diversity of the microbiota from air and surface samples aboard ISS increased from an initial six to 53 different bacterial species. During the same period, fungal diversity increased from 2 to 24 species. Based upon rep-PCR analysis, the majority of isolates were unique suggesting the need for increased sampling frequency and a more thorough analysis of samples to properly characterize the ISS microbiota. This limited fungal and bacterial data from environmental samples acquired during monitoring currently do not indicate a microbial hazard to ISS or any trends suggesting potential health risks.

Fontenot, Sondra L.↗

Effect Of Spaceflight On Microbial Gene Expression And Virulence: Preliminary Results From Microbe Payload Flown On-Board STS-115

Human presence in space, whether permanent or temporary, is accompanied by the presence of microbes. However, the extent of microbial changes in response to spaceflight conditions and the corresponding changes to infectious disease risk is unclear. Previous studies have indicated that spaceflight weakens the immune system in humans and animals. In addition, preflight and in-flight monitoring of the International Space Station (ISS) and other spacecraft indicates the presence of opportunistic pathogens and the potential of obligate pathogens. Altered antibiotic resistance of microbes in flight has also been shown. As astronauts and cosmonauts live for longer periods in a closed environment, especially one using recycled water and air, there is an increased risk to crewmembers of infectious disease events occurring in-flight. Therefore, understanding how the space environment affects microorganisms and their disease potential is critically important for spaceflight missions and requires further study. The goal of this flight experiment, operationally called MICROBE, is to utilize three model microbial pathogens, Salmonella typhimurium, Pseudomonas aeruginosa, and Candida albicans to examine the global effects of spaceflight on microbial gene expression and virulence attributes. Specifically, the aims are (1) to perform microarray-mediated gene expression profiling of S. typhimurium, P. aeruginosa, and C. albicans, in response to spaceflight in comparison to ground controls and (2) to determine the effect of spaceflight on the virulence potential of these microorganisms immediately following their return from spaceflight using murine models. The model microorganisms were selected as they have been isolated from preflight or in-flight monitoring, represent different degrees of pathogenic behavior, are well characterized, and have sequenced genomes with available microarrays. In particular, extensive studies of S. typhimurium by the Principal Investigator, Dr. Nickerson, using ground-based analog systems demonstrate important changes in the genotypic, phenotypic, and virulence characteristics of this pathogen resulting from exposure to a flight-like environment (i.e. modeled microgravity).

Wilson, J. W.↗

The Validation of Vapor Phase Hydrogen Peroxide Microbial Reduction for Planetary Protection and a Proposed Vacuum Process Specification

The Jet Propulsion Laboratory, in conjunction with the NASA Planetary Protection Officer, has selected the vapor phase hydrogen peroxide sterilization process for continued development as a NASA approved sterilization technique for spacecraft subsystems and systems. The goal is to include this technique, with an appropriate specification, in NPR 8020.12C as a low temperature complementary technique to the dry heat sterilization process.To meet microbial reduction requirements for all Mars in-situ life detection and sample return missions, various planetary spacecraft subsystems will have to be exposed to a qualified sterilization process. This process could be the elevated temperature dry heat sterilization process (~115 C for 40 hours) which was used to sterilize the Viking lander spacecraft. However, with utilization of such elements as highly sophisticated electronics and sensors in modern spacecraft, this process presents significant materials challenges and is thus an undesirable bioburden reduction method to design engineers. The objective of this work is to introduce vapor hydrogen peroxide (VHP) as an alternative to dry heat microbial reduction to meet planetary protection requirements.The VHP process is widely used by the medical industry to sterilize surgical instruments and biomedical devices, but high doses of VHP may degrade the performance of flight hardware, or compromise material properties. Our goal for this study was to determine the minimum VHP process conditions to achieve microbial reduction levels acceptable for planetary protection.

planetary protection↗

Development of an Autofluorescence Spectral Database for the Identification and Classification of Microbial Extremophiles

Extremophiles are microorganisms that have adapted to severe conditions that were once considered devoid of life. The extreme settings in which these organisms flourish on earth resemble many extraterrestrial environments. Identification and classification of extremophiles in situ (without the requirement for excessive handling and processing) can provide a basis for designing remotely operated instruments for extraterrestrial life exploration. An important consideration when designing such experiments is to prevent contamination of the environments. We are developing a reference spectral database of autofluorescence from microbial extremophiles using long-UV excitation (405 nm). Aromatic compounds are essential components of living systems, and biological molecules such as aromatic amino acids, nucleotides, porphyrins and vitamins can also exhibit fluorescence under long-UV excitation conditions. Autofluorescence spectra were obtained from a confocal microscope that additionally allowed observations of microbial geometry and motility. It was observed that all extremophiles studied displayed an autofluorescence peak at around 470 nm, followed by a long decay that was species specific. The autofluorescence database can potentially be used as a reference to identify and classify past or present microbial life in our solar system.

Sabanayagam, Chandran↗

Characterization of Volume F Trash from Four Recent STS Missions: Microbial Occurrence, Numbers, and Identifications

The fate of space-generated solid wastes, including trash, for future missions is under consideration by NASA. Several potential treatment options are under active technology development. Potential fates for space-generated solid wastes: Storage without treatment; storage after treatment(s) including volume reduction, water recovery, sterilization, and recovery plus recycling of waste materials. For this study, a microbial characterization was made on trash returned from four recent STS missions. The material analyzed were 'Volume F' trash and other bags of accompanying trash. This is the second of two submitted papers on these wastes. This first one covered trash content, weight and water content. Upon receipt, usually within 2 days of landing, trash contents were catalogued and placed into categories: drink containers, food waste, personal hygiene items, and packaging materials, i.e., plastic film and duct tape. Microbial counts were obtained with cultivatable counts on agar media and direct counts using Acridine Orange fluorescent stain (AODC). Trash bag surfaces, 25 square cm , were also sampled. Direct counts were approximately 1 x 10(exp 6) microbes/square cm and cultivatable counts ranged from 1 x 10 to 1 X 10(exp 4) microbes/ square cm-2. Aerobic microbes, aerobic sporeformers, and yeasts plus molds were common for all four missions. Waste items from each category were placed into sterile ziplock bags and 1.5 L sterile DI water added. These were then dispersed by hand shaking for 2 min. prior to inoculation of count media or determining AODC. In general, cultivatable microbes were found in drinks, food wastes, and personal hygiene items. Direct counts were usually higher than cultivatable counts. Some pathogens were found: Staphylococcus auerus, Escherichia coli (fecal wastes). Count ranges: drink pouches - AODC 2 x 10(exp 6) to 1 X 10(exp 8) g(sub fw) (exp -1); cultivatable counts variable between missions; food wastes: Direct counts were close to aerobic plate counts. Counts ranged from 10(exp 6) to 10(exp 9) per g(sub fw). Identities of isolates from cultivation media were obtained using a Biolog Microbial ID System or microSEQ molecular ID methodology using an ABI3130 gene analyzer.

Strayer, Richard F.↗

Marine Microbial Mats and the Search for Evidence of Life in Deep Time and Space

Cyanobacterial mats in extensive seawater evaporation ponds at Guerrero Negro, Baja California, Mexico, have been excellent subjects for microbial ecology research. The studies reviewed here have documented the steep and rapidly changing environmental gradients experienced by mat microorganisms and the very high rates of biogeochemical processes that they maintained. Recent genetic studies have revealed an enormous diversity of bacteria as well as the spatial distribution of Bacteria, Archaea and Eukarya. These findings, together with emerging insights into the intimate interactions between these diverse populations, have contributed substantially to our understanding of the origins, environmental impacts, and biosignatures of photosynthetic microbial mats. The biosignatures (preservable cells, sedimentary fabrics, organic compounds, minerals, stable isotope patterns, etc.) potentially can serve as indicators of past life on early Earth. They also can inform our search for evidence of any life on Mars. Mars exploration has revealed evidence of evaporite deposits and thermal spring deposits; similar deposits on Earth once hosted ancient microbial mat ecosystems.

Des Marais, David J.↗

Microbial Monitoring Challenges and Needs for Mars Applications

The monitoring of microorganisms will be an important part of a mission to Mars. Microbial monitoring equipment will be needed to look for the presence of microorganisms on the planet, to confirm that planetary protection measures are working, to monitor the health of plants, bioreactors and humans living in a habitat and to monitor the performance of the life support systems that will keep them alive during their stay on Mars. Coordinating the different microbial monitoring needs during the early days of mission planning, can provide NASA with equipment that could meet more than one need while also providing complementary analysis options, which can enhance the research capabilities. The early coordination between the different NASA groups that will need microbial monitoring equipment on the surface of Mars, could also make the mission more affordable, as development of the needed equipment could be potentially cost shared.

Roman, M. C.↗

Development of a Centrifugal Technique for the Microbial Bioburden Analysis of Freon (CFC-11)

NASA Procedural Requirement 8020.12C entitled "Planetary Protection Provisions for Robotic Extraterrestrial Missions" states that the source-specific encapsulated microbial density for encapsulated organisms (div(0)) in nonmetallic materials ranges from 1-30 spores/cubic cm. The standard laboratory procedure, NASA Standard Procedures for the Microbial Examination of Space Hardware, NHB 5340.1B, does not provide any direction into the methodologies to understand the bioburden within such a fluid as CFC-11 (Freon). This general specification value for the Freon would be applicable to the Freon charged within the Mars Science Laboratory fs (MSL fs) Heat Rejection System. Due to the large volume required to fill this system, MSL could not afford to conservatively allocate 55.8% of the total spore budget of the entire laboratory system (rover, descent stage, cruise stage, and aeroshell) of 5.00 X 10(exp 5) spores at launch. A novel filtration approach was developed to analyze the Freon employing a 50 kDa molecular weight cutoff (MCO) filter, followed by 0.22-micron pore-size filter to establish a calculated microbial bioburden.

Benardini, James N.↗

Microbial Monitoring of Common Opportunistic Pathogens by Comparing Multiple Real-time PCR Platforms for Potential Space Applications

The International Space Station is a closed environment with rotation of astronauts and equipment that each introduce their own microbial flora, it is necessary to monitor the air, surfaces, and water for microbial contamination. Current microbial monitoring includes labor- and time-intensive methods to enumerate total bacterial and fungal cells, with limited characterization, during in-flight testing. Although this culture-based method is sufficient for monitoring the International Space Station, on future long-duration missions more characterization will need to be performed during flight, as sample return and ground characterization may not be available.

Real time↗

In-Flight Microbial Monitor

Previous research has shown that potential human pathogens have been detected on the International Space Station (ISS). New microorganisms are introduced with every exchange of crew and cargo. Microorganisms introduced to the ISS are readily transferred between crew and subsystems (i.e., ECLSS, environmental control and life support systems). Current microbial characterization methods require a culture-based enrichment of microorganisms and at least a 48-hour incubation time. This increases the microbial load while detecting only a limited number of microorganisms. The culture-based method detects approximately 1-10% of the total organisms present and provides no identification. To identify and enumerate ISS samples requires that the microbes be returned to Earth for complete analysis. Therefore, a more expedient, low-cost, inflight method of microbial detection, identification, and enumeration is needed. The RAZOR EX, a ruggedized, commercial off the shelf, real-time PCR field instrument was tested for its ability to detect microorganisms at low concentrations within one hour. Escherichia coli, Salmonella enterica Typhimurium, and Pseudomonas aeruginosa were detected at low levels using real-time DNA amplification. Total heterotrophic counts could also be detected using a 16S gene marker that can identify up to 98% of all bacteria. To reflect viable cells found in the samples, RNA was also detectable using a modified, single-step reverse transcription reaction.

Technology Portfolio System↗