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At least 181 records · Page 10

Effects of aeolian erosion on microbial release from solids.

This study was initiated to determine the percentage of spores that would be expected to be released from the interior of solid materials by aeolian erosion on a planetary surface. Methyl methacrylate and Eccobond disks were fabricated so that each disk contained approximately 40,000 Bacillus subtilis var. niger spores. The disks were placed in a specially designed sandblasting device and eroded. Exposure periods of 0.5, 2 and 24 hours were investigated using filtered air to accelerate the sand. A series of tests was also conducted for a 0.5 hour period using carbon dioxide. Examination of the erosion products showed that less than 1% of the spores originally contained in the solids was released by aeolian erosion.

Gustan, E. A.↗

Quantitation of buried contamination by use of solvents

An investigation was made to determine (1) sporicidal properties of amine solvents that solubilize silicon resins, (2) recovery properties of a silicon potting compound (RTV 41) used in spacecraft, and (3) viability of spores during chemical curing of the potting compound. Results show that: (1) spores do remain viable during RTV 41 silicon potting chemical curing, and (2) spore recovery from cured silicon potting compound RTV 41 is very high when silicon rubber is dissolved in butylamine and series dilution with benzene prior to plate curing.

Pappas, S. P.↗

Quantitation of buried contamination by use of solvents

A method for determining the quantity of buried contamination using solvents is presented. A nonsporocidal method with which high spore recoveries are achievable from silicone coatings and potting compounds was developed. An extension of the method to silicon potting compound RTV 60 is reported. It is stated that spores remain viable during chemical curing of silicone potting compounds and more than ninety percent of the spore population is recoverable by amine dissolution and proper plating techniques.

Pappas, S. P.↗

Environmental microbiology as related to planetary quarantine

The survival of Bacillus subtilis var. niger spores suspended in solutions of sucrose and glycerol at calculated water activities and varying temperatures was studied. The overall results indicated that as the water activity of the liquid decreased from .99 to .85, the heat resistance of the spores increased. The nature of the substance controlling the water activity, and the history of the spores prior to treatment also had an affect on their heat resistance.

Pflug, I. J.↗

Ecology and thermal inactivation of microbes in and on interplanetary space vehicle components

Dry heat inactivation characteristics were compared for 4-6 (B. brevis) spores and microbes from the Cincinnati soil samples at 105, 112, and 125. Characterized were the survival curves of 4-6 (B. brevis) spores at 112, 115, 118, 120, and 125 C, and 1.2 microgram of water per ml of headspace air (closed tin-can system), and the morphological characteristics of 4-6 (B. brevis), 6-12 (B. lentus), 7-11 (B. coagulans), and B. subtilis var. niger spores by scanning electron microscopy.

Reyes, A. L.↗

Growth-rate periodicity of Streptomyces levoris during space flight

Streptomyces levoris provides a suitable biological test system to investigate the effects of space flight on the rhythms of vegetative and spore phase characteristics of both growth-rate periodicity and culture morphology during the pre-, in-, and post-flight periods of the Apollo-Soyuz Test Project. The objectives of the American participation were to study the effects of space flight on the biorhythms of Streptomyces levoris based on a comparison of the growth-rate periodicity of the vegetative and spore phase within each culture, to examine the possible alteration of spore morphology and development by SEM, and to compare the effects of a 12-hr phase shift on the periodic growth characteristics of this microorganism in cultures which were exchanged during the joint activities of the space flight. No uniform differences in the biorhythm of Streptomyces levoris during space flight were observed. It appears that the single most variable factor related to the experiment was the lack of temperature control for the space-flight specimens.

Rogers, T. D.↗

Dry-heat resistance of selected psychrophiles

The dry-heat resistance characteristics of spores of psychrophilic organisms isolated from soil samples from the Viking spacecraft assembly areas at Cape Kennedy Space Flight Center, Cape Canaveral, Fla., were studied. Spore suspensions were produced, and dry-heat D values were determined for the microorganisms that demonstrated growth or survival under a simulated Martian environment. The dry-heat tests were carried out by using the planchet-boat-hot plate system at 110 and 125 C with an ambient relative humidity of 50% at 22 C. The spores evaluated had a relatively low resistance to dry heat. D (110 C) values ranged from 7.5 to 122 min, whereas the D (125 C) values ranged from less than 1.0 to 9.8 min.

Winans, L.↗

Photoabsorption Study of Bacillus megaterium, DNA and Related Biological Materials in the Phosphorus K-edge Region

We have measured the x-ray transmission spectra of several biologically related samples in the phosphorus K-edge absorption region. These include elemental red phosphorus, hydrated sodium phosphate (Na3PO4.12H2O), deoxyribonucleic acid (DNA), adenosinetriphosphate (ATP), diolylphosphatidyl choline (DOPC), and Bacillus megaterium spores. Elemental red phosphorus essentially displays an edge-jump. All other spectra are similar in form and energy position. Each spectrum for these substances is dominated by a narrower, more intense first peak and a broader but less intense second peak. The corresponding K-edge absorption thresholds are shifted towards higher energy relative to that for elemental red phosphorus, as expected for increasing degrees of phosphorus oxidation. The B. megaterium spectrum has aspects common to both the phosphate and DNA spectra and is therefore interpreted as a composite of spectra arising from DNA/RNA and phosphates within the spore. The B. megaterium spore spectrum provides needed information for resonant radiation damage studies in the phosphorus K-edge absorption region by identifying candidate photoexcitations. In addition,the absorption spectra will be useful in macromolecular crystallography studies employing anomalous dispersion effects at the phosphorus K-edge.

Frigo, Sean P.↗

Photoabsorption study of Bacillus megaterium, DNA and Related Biological Materials in the Phosphorus K-edge Region

We have measured the x-ray transmission spectra of several biologically related samples in the phosphorus K-edge absorption region. These include red phosphorus, hydrated sodium phosphate (Na3PO4 12 H2O), deoxyribonucleic acid (DNA), adenosinetriphosphate (ATP), diolylphosphatidyl choline (DOPC), and Bacillus megaterium spores. Red phosphorus essentially displays an edge-jump. All other spectra are similar in form and energy position, where each is dominated by a narrower, more intense first peak and a broader but less intense second peak. The corresponding K-edge absorption thresholds are shifted towards higher energy relative to that for red phosphorus, as expected for increasing degrees of phosphorus oxidation. The B.meguterium spectrum has aspects common to both the phosphate and DNA spectra and is therefore interpreted as a composite of spectra arising from DNA/RNA and phosphates within the spore. The B. megaterium spore spectrum provides needed information for resonant radiation damage studies in the phosphorus K-edge absorption region by identifying candidate photoexcitations. In addition, the absorption spectra will be useful in macromolecular crystallography studies employing anomalous dispersion effects at the phosphorus K-edge.

Frigo, Sean P.↗

Arthromitus (Bacillus cereus) symbionts in the cockroach Blaberus giganteus: dietary influences on bacterial development and population density

The filamentous spore-forming bacterium Arthromitus, discovered in termites, millipedes, sow bugs and other soil-dwelling arthropods by Leidy (1850), is the intestinal stage of Bacillus cereus. We extend the range of Arthromitus habitats to include the hindgut of Blaberus giganteus, the large tropical American cockroach. The occurrence and morphology of the intestinal form of the bacillus were compared in individual cockroaches (n=24) placed on four different diet regimes: diurnally maintained insects fed (1) dog food, (2) soy protein only, (3)purified cellulose only, and (4) a dog food-fed group maintained in continuous darkness. Food quality exerted strong influence on population densities and developmental stages of the filamentous bacterium and on fecal pellet composition. The most dramatic rise in Arthromitus populations, defined as the spore-forming filament intestinal stage, occurred in adult cockroaches kept in the dark on a dog food diet. Limited intake of cellulose or protein alone reduced both the frequency of Arthromitus filaments and the rate of weight gain of the insects. Spores isolated from termites, sow bugs, cockroaches and moths, grown on various hard surfaces display a branching mobility and resistance to antibiotics characteristic to group I Bacilli whose members include B. cereus, B. circulans, B. alvei and B. macerans. DNA isolated from pure cultures of these bacilli taken from the guts of Blaberus giganteus (cockroach), Junonia coenia (moth), Porcellio scaber (sow bug) and Cryptotermes brevis (termite) and subjected to Southern hybridization with a 23S-5S B. subtilis ribosomal sequence probe verified that they are indistinguishable from laboratory strains of Bacillus cereus.

NASA Discipline Exobiology↗

Polar distribution of annexin-like proteins during phytochrome-mediated initiation and growth of rhizoids in the ferns Dryopteris and Anemia

Although the calcium requirement of phytochrome-mediated fern spore germination and early rhizoid growth is well established, the calcium-binding proteins that serve as transducers for these responses are not known. Here we report the presence of annexin-like proteins in germinating spores of Dryopteris filix-mas (L.) Schott and Anemia phyllitidis (L.) Sw. and evidence that they may be important participants in early photomorphogenic changes in gametophytes. Immunolocalization and immunoblot assays of these proteins were carried out using polyclonal antibodies raised either against a 35-kDa annexin-like protein from pea or against anchorin CII from chicken. Western-blot analysis showed that crude protein extracts obtained from both species after red-light treatment contained two cross-reactive protein bands with molecular weights around 70 kDa. These proteins were annexin-like in that they bound to a phosphatidylserine affinity column in a calcium-dependent fashion. Using this column, two protein bands around 70 kDa, i.e. 67 and 73 kDa, were partially purified together with proteins at 36 kDa and a doublet at 54 kDa. Proteins of these latter molecular weights are suggested to be members of the annexin family, but no cross-reactivity could be found between these and the two antibodies used in our investigations. Immunodetectable levels of these proteins were observed only after light-mediated induction of spore germination. Imaging of the immuno-localization patterns observed with both antibodies showed that the annexin-like proteins are concentrated at the extreme tips of the rhizoids in D. filix-mas and A. phyllitidis during rhizoid initiation and all stages of elongation. We suggest that these proteins may play a major role in the tip-oriented exocytosis events that are critical for the initiation and growth of fern rhizoids.

NASA Discipline Number 40-50↗

Phylogeny of marine Bacillus isolates from the Gulf of Mexico

The phylogeny of 11 pigmented, aerobic, spore-forming isolates from marine sources was studied. Forty-two biochemical characteristics were examined, and a 16S rDNA sequence was obtained for each isolate. In a phylogenetic tree based on 16S sequencing, four isolates (NRRL B-14850, NRRL B-14904, NRRL B-14907, and NRRL B-14908) clustered with B. subtilis and related organisms; NRRL B-14907 was closely related to B. amyloliquefaciens. NRRL B-14907 and NRRL B-14908 were phenotypically similar to B. amyloliquefaciens and B. pumilus, respectively. Three strains (NRRL B-14906, NRRL B-14910, and NRRL B-14911) clustered in a clade that included B. firmus, B. lentus, and B. megaterium. NRRL B-14910 was closely related phenotypically and phylogenetically to B. megaterium. NRRL B-14905 clustered with the mesophilic round spore-producing species, B. fusiformis and B. sphaericus; the isolate was more closely related to B. fusiformis. NRRL B-14905 displayed characteristics typical of the B. sphaericus-like organisms. NRRL B-14909 and NRRL B-14912 clustered with the Paenibacillus species and displayed characteristics typical of the genus. Only NRRL B-14851, an unusually thin rod that forms very small spores, may represent a new Bacillus species.

NASA Discipline Exobiology↗

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

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

Strayer, Richard F.↗

Microbial Characterization Space Solid Wastes Treated with a Heat Melt Compactor

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

Strayer, Richard F.↗

Analysis of CO2 Composite Spray Cleaning System Results

In a previous study, the cleaning efficiency of a CO2 composite cleaning system for particulate removal was tested. The study covered particles from spores to fluorescent particles of different sizes as well as a variety of substrate surfaces, including aluminum, titanium, stainless steel, and nitinol. Particles were deposited using aerosol (dry) and droplet (wet) deposition. Results from the previous study show that the CO2 composite spray system is capable of cleaning to sterility for aerosol deposited spores and is capable of cleaning a minimum of a 4-log reduction for droplet deposited spores. This minimum 4-log reduction matches current Planetary Protection dry heat microbial reduction requirements. In this paper we will present new data to further correlate the cleaning efficiency with how contamination was introduced to the surface, the surface roughness, and particle size. Possible causes for such correlations will be discussed.

Chung, Shirley↗

Microbial Characterization of Heat Melt Compaction for Treatment of Space Generated Solid Wastes

One treatment process in development for solid waste management in space has been the Trash Compaction Processing System (TCPS). Human space mission wastes typically contain large percentages of contaminated wet solid waste. The Heat Melt Compactor (HMC) is being developed to be a multi-function means of water recovery, volume reduction, and the making safe of contaminant-rich trash with the potential for waste stabilization and/or sterilization. To determine the efficacy of the HMC treatment to kill microorganisms in solid waste and remain biologically stable, testing was conducted on three tiles produced by HMC-Gen 2 at Ames Research Center. Samples were shipped to Kennedy Space Center to test for microbial viability after compaction, determine the bio-stability of the HMC disks during storage (43 days), and assess potential airborne contaminate microbial growth on surfaces at low and high humidity conditions. In addition to the products of solid waste processing technologies, there is a concern that the crew might come into contact with hardware surfaces that have been contaminated by microorganisms during waste processing. The extent of microbial surface contamination of waste processing hardware was determined by surface sample swabbing and analysis for total bacterial and yeast counts and cultivable counts of aerobic and anaerobic bacteria, spore-forming bacteria, and fungi. Results indicate that trash processing increased bacterial counts on the surfaces of the compacter. All but one biological indicator spore strip imbedded in the HMC produced tiles were negative for growth after incubation for five days indicating effective sterilization through the heat melt compaction process. Analysis of core samples as well as surface growth of tiles inoculated with Aspergillus niger fungal spores incubated at three levels of humidity indicate that HMC created tiles did not support the proliferation of bacterial and fungal growth.

Mary E Hummerick↗

Unlocking the Spacecraft and Human Habitat Microbiome to Enable the Next Generation of Space Exploration

Planetary protection is the discipline that prevents harmful contamination of the solar system during exploration activities. The current international guidelines and NASA policy addressing biological contamination on spacecraft surfaces contains prescriptive guidelines of spore requirements (e.g., 300 spores/m2, 5×105 spores per spacecraft) applicable to spacecraft bound for Mars. To verify these requirements spacecraft engineers sample spacecraft surfaces throughout the assembly, test and launch operations phase of the mission using damp water cotton swabs and polyester wipes. After sampling, the potential biological contamination is enumerated using a series of traditional microbiology techniques to include sonication, heat shocking at 80°C for 15min to select for spores, and growth on tryptic soy agar at 32°C for 72 hours. To enable crewed missions to Mars and robotic exploration of the Ocean Worlds a risk informed decision making / performance-based approach to assess biological contamination offers a promising solution in the trade space. Recognizing the need for a performance-based approach, NASA’s new Planetary Protection policies now incorporate the agility for missions to be able to leverage a performance or prescriptive approach. One of top contenders in the option space is a coupled quantitative, descriptive and functional based approach to be able to assess the quantity, types and capabilities of the biological contamination present on spacecraft surfaces. A tailored, mission by mission assurance case could then be formulated by building an argument around the target body, projected capabilities surrounding the types of organisms their potential for survival and proliferation, and ability to be transported on the target body to contaminate an area of biological interest. A performance-based requirement would then be used to demonstrate the mission’s compliance in protecting the planetary environment safety objectives. This symposium talk will showcase the background and need case for NASA to develop such a capability as well as provide an update on the efforts underway in developing a transparent and responsible performance-based approach to biological contamination assessments on spacecraft surfaces.

Habitat Microbiome↗

Molecular microbial diversity of a spacecraft assembly facility

In ongoing investigations to map and archive the microbial footprints in various components of the spacecraft and its accessories, we have examined the microbial populations of the Jet Propulsion Laboratory's Spacecraft Assembly Facility (JPL-SAF). Witness plates made up of spacecraft materials, some painted with spacecraft qualified paints, were exposed for approximately 7 to 9 months at JPL-SAF and examined the particulate materials collected for the incidence of total cultivable aerobic heterotrophs and heat-tolerant (80 degrees C for 15-min.) spore-formers. The results showed that the witness plates coated with spacecraft qualified paints attracted more dust particles than the non-coated stainless steel witness plates. Among the four paints tested, witness plates coated with NS43G accumulated the highest number of particles, and hence attracted more cultivable microbes. The conventional microbiological examination revealed that the JPL-SAF harbors mainly Gram-positive microbes and mostly spore-forming Bacillus species. Most of the isolated microbes were heat resistant to 80 degrees C and proliferate at 60 degrees C. The phylogenetic relationships among 23 cultivable heat-tolerant microbes were examined using a battery of morphological, physiological, molecular and chemotaxonomic characterizations. By 16S rDNA sequence analysis, the isolates fell into seven clades: Bacillus licheniformis, B. pumilus, B. cereus, B. circulans, Staphylococcus capitis, Planococcus sp. and Micrococcus lylae. In contrast to the cultivable approach, direct DNA isolation, cloning and 16S rDNA sequencing analysis revealed equal representation of both Gram-positive and Gram-negative microorganisms.

Industry↗