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At least 127 records · Page 7

Exploring Sustained Food Production in the Veggie Vegetable Production System

Since 2014, the Veggie vegetable production system on the International Space Station has intermittently supplied astronaut crews with fresh produce. To assess the potential for continuous crop production in Veggie and develop a baseline for future space crop production systems, a 120-day study was conducted to determine methodology for inputs, optimal yield, food safety standards, and crew involvement. ‘Amara’ mustard and ‘Red Russian’ kale were grown as initial crops, followed by ‘Extra Dwarf’ pakchoi and shungiku as final crops. Previous grow-outs in Veggie have included harvests at 28-35 days after initiation. In this study, a 56-day grow-out with multiple harvests from the same plants was compared to the conventional, single harvest Veggie schedule. Unlike previous Veggie studies which grew all plants simultaneously, this test staggered initiation and harvest, aiming for consistent and increased production. Plant pillows were initiated in pairs weekly and positioned to reduce shading. Completed pillows were immediately replaced with fresh ones. The multi-harvest scheme used fewer pillows, totaling 46% less pillow mass than the single harvest method. Yield varied by crop and harvest scheme, when compared as fresh edible biomass production across 56-day time increments. ‘Red Russian’ kale yielded similarly across harvest schemes. In the multiple-harvest schedule, ‘Amara’ mustard and shungiku yielded 23% to 25% higher, respectively, while ‘Extra Dwarf’ pakchoi had 43% lower yield. Microbial analysis of the plants indicated no culturable human pathogens. Microbial load of a given plant appeared to depend more on system age than plant age; across harvest methods, aerobic plate counts from final crops were higher than those of initial crops. This project also considered the complexity of crew involvement in a continuous production scenario. New crew procedures that periodically remove plant material from the Veggie root mat would be needed under continuous production to prevent potential pathogens and unpleasant odors. This study supports future space crop production scenarios and was funded by NASA’s Human Research Program.

Jess M Bunchek↗

Spaceflight-Induced Changes in Microbial Virulence and the Impact to the Host Immune Response

Over the past 50 years, microorganisms have displayed unexpected responses relevant to infectious disease when grown in spaceflight and spaceflight analogues, including changes in final cell concentration, biofilm production, stress resistance, antibiotic sensitivity, gene expression, and virulence. •Seminal studies demonstrated that the foodborne pathogen, Salmonella enterica serovar Typhimurium, increased its virulence and pathogenesis-related characteristics in response to both spaceflight and spaceflight analogue culture. Since those experiments, alterations in the pathogenesis-related characteristics of other pathogens have been documented in response to growth in these environments4,5,6, and a recent study has demonstrated an increase in virulence of Serratia marcescens cultured during spaceflight7. However, our overall knowledge of which microorganisms will alter their virulence in response to spaceflight and spaceflight analogue culture remains very limited

C. M. Ott↗

Microbial Characteristics of ISS Environmental Surfaces

The microbiome of environmental surfaces from the International Space Station were characterized in order to examine the relationship to crew and hardware maintenance. The Microbial Observatory (ISS-MO) experiment generated a microbial census of ISS environments using advanced molecular microbial community analyses along with traditional culture-based methods. Since the “omics” methodologies generated an extensive microbial census, significant insights into spaceflight-induced changes in the populations of beneficial and/or potentially harmful microbes were gained. Surface samples were collected from several ISS surface locations from three flight opportunities, and were returned to Earth via the Soyuz TMA-14M or the Space X Dragon capsule. In addition to cultivation methods, viable microbial burden, iTag-based sequencing, and metagenome analyses were carried out. The cultivable microbial bioburden differed by location and sampling event. Exploring the ISS environmental microbiome revealed presence of opportunistic pathogens and antibiotic resistant microbes. Genes involved in ATP binding cassette transporters, two component systems, and beta-lactam resistance were among a diverse set of metabolic and genetic information processing pathways. Whole genome sequencing (WGS) of 50 ISS strains exhibiting resistance to various antibiotics was carried out. The antibiotic resistant genes deduced from the WGS were compared with the resistomes generated directly from the gene pool of the environmental samples. Two unique Aspergillus fumigatus strains isolated from the ISS were characterized and compared to the experimentally established clinical isolates Af293 and CEA10. A virulence assessment in a neutrophil-deficient larval zebrafish model of invasive aspergillosis indicated that both ISSFT-021 and IF1SW-F4 were significantly more lethal compared to Af293 and CEA10. The findings from this Environmental “Omics” project should be exploited to enhance human health and well-being of a closed system. In other words, the ISS-MO research aims to "translate" findings in fundamental research into medical practice (pathogen detection) and meaningful health outcomes (countermeasure development).

Perry, Jay↗

Sustained Veggie: A Continuous Food Production Comparison

The International Space Station’s Veggie system intermittently supplies the crew with fresh produce. To assess the potential for continuous crop production in Veggie and develop a baseline for future space crop production systems, a 120-day study was conducted to determine methodology for inputs, optimal yield, and crew involvement. ‘Amara’ mustard and ‘Red Russian’ kale were grown as initial crops, followed by ‘Extra Dwarf’ pak choi and shungiku as final crops. Previous grow-outs in Veggie have included harvests at 28-35days after initiation. In this study, a 56-day grow-out with multiple harvests from the same plants was compared to the conventional, single harvest Veggie schedule. Unlike previous Veggie studies which grew all plants simultaneously, this test staggered initiation and harvest, aiming for consistent and increased production. Plant pillows were initiated in pairs weekly and positioned to reduce shading. Completed pillows were immediately replaced with fresh ones. The multi-harvest scheme used fewer pillows, totaling 46% less pillow mass than the single harvest method. Scaled to 56-day increments, yield varied by crop and harvest scheme. ‘Red Russian’ kale yielded similarly across harvest schemes. In the multiple-harvest schedule, ‘Amara’ mustard and shungiku yielded 23% to 25% higher, respectively, while ‘Extra Dwarf’ pak choi had43% lower yield. Microbial analysis of the plants indicated no culturable human pathogens. Microbial load of a given plant appears to depend more on system age than plant age. Across harvest methods, aerobic plate counts from final crops were higher than those of initial crops. This project also considered the complexity of crew involvement in a continuous production scenario. New crew procedures that periodically remove plant material from the Veggie root mat are needed under continuous production to prevent potential pathogens and unpleasant odors. This study supports future space crop production scenarios and was funded by NASA’s Human Research Program.

Jess M Bunchek↗

Enterococci in Space: Adaptation, Antibiotic Resistance, and Clinical Implications

Enterococci are gram-positive bacteria that originated when our ancient animal ancestors emerged from the oceans to live on land, and brought their gut flora with them. Enterococcus faecalis (EF) and Enterococcus faecium, are common human commensals and can harbor multidrug resistance. Both have been previously isolated from the International Space Station (ISS). Likely as a consequence of their evolutionary origins, enterococci show remarkable stress resistance within, but also outside, their human hosts. Their antibiotic resistance, coupled with tolerance to desiccation, starvation, and disinfection, make some EF strains potent pathogens in the built environment (e.g., hospitals), and a potential risk to crew health during space missions. Here we describe our planned flight studies, currently in development. Genomic Enumeration of Antibiotic Resistance in Space (GEARS) will characterize the frequency and genomic identity of antibiotic resistant organisms, including enterococci, on the ISS, and expand our future in-space sequencing-based diagnostic capabilities. Enterococcus Growth Advantage on ISS via Tn-seq (EnteroGAIT) will assess the evolutionary selective pressure of the space environment (microgravity, space radiation) using EF as a model system during a long-duration culture and persistence experiment. Adaptation & Evolution of Resilient Enterococcus in Space (AERES) will study existing and newly identified isolates to characterize the “natural” evolutionary history of EF on Earth and in space to reveal mechanisms of microbial adaption including, possibly, natural selection. Our work to date has identified that EF ISS isolates share virulence factors found in clinical strains and commensals, and revealed limitations of current pathogenicity assessment tools. While ISS isolates were, in some cases, multi-drug resistant, current data suggests these are largely commensal strains. Our work will further refine potential crew health risks and improve understanding of EF adaptation to the built environment, of great relevance on Earth where EF is the second leading cause of hospital acquired infection.

Jordan McKaig↗

Effects of Spaceflight Relevant Carbon Dioxide Levels on Pathogenesis Related Microbial Characteristics

BACKGROUND Multiple stressors in the spaceflight environment have the potential to alter microbial pathogenesis and virulence characteristics, including microgravity, radiation, and hostile/confined conditions. This combination of factors may lead to a stacking of risks, potentially creating unexpected increase in the risk of infectious disease. One understudied variable is the higher CO2 atmospheric concentration. The current Spaceflight Maximum Allowance Concentration (SMAC) for 24-hour average CO2 is 0.4% (3 mm Hg). These elevated ambient CO2 levels aboard the ISS could potentially influence the diversity and phenotypic responses of the resident microbial communities from both the spacecraft environment (air, surface, water) and crewmembers (gut, nasal, skin microbiomes). METHODS For these studies, we will evaluate the response of Staphylococcus aureus, Streptococcus pneumoniae and Enterobacter aerogenes, which are opportunistic pathogens that are of medical significance and have been or are likely to be found aboard spacecraft. These microorganisms will be cultured in both spaceflight analog and control conditions at CO2levels representing terrestrial atmospheric concentration of 0.04% (0.3 mmHg), ISS elevated concentration of 0.4% (3 mm Hg), and an unexpected elevated concentration of 1 % (7.6 mmHg) to represent a worst case scenario. DISCUSSION Overwhelming evidence from studies performed in food microbiology, marine biology and terrestrial environmental biology suggest altered gene expression, selective bacterial inhibition, increased growth and diversity, and increased antibiotic resistance of bacterial communities individually and in biofilm formation when exposed to increased levels of CO2. While many of these findings are based on levels of CO2 higher than would be found during spaceflight exploration missions, the potential for subtle changes in CO2 levels to exacerbate infectious disease risks warrants an evaluation of pathogen responses to spaceflight CO2 conditions. Our study will provide answers to spaceflight relevant atmospheric conditions, as described in the Risk of Adverse Health Effects Due to Host-Microorganism Interaction (GapMicro 103).

A A Medina-Colorado↗

Effects of Spaceflight Relevant Carbon Dioxide Levels on Pathogenesis Related Microbial Characteristics

Multiple stressors in the spaceflight environment, including microgravity and radiation, have the potential to alter microbial pathogenesis and virulence characteristics. Understanding how these and other factors may exacerbate the risk of infectious disease for astronauts is important to ensure the success of the mission. One understudied question in our assessment of infectious disease risk is the contribution carbon dioxide (CO2) at atmospheric concentrations higher than would be observed in terrestrial settings. Previous terrestrial studies have shown altered gene expression, selective bacterial inhibition, increased growth and diversity, and increased antibiotic resistance of bacterial communities individually and in biofilms when exposed to increased levels of CO2. While many of these findings are based on levels of CO2 higher than would be found during spaceflight exploration missions, the potential for subtle changes in CO2 levels to exacerbate infectious disease risks warrants an evaluation of pathogen responses to spaceflight CO2 conditions. To address this question, we are evaluating the effects of elevated atmospheric CO2 on microbial concentration, diversity, and pathogenesis-related characteristics using medically significant microorganisms cultured at CO2 levels representing terrestrial atmospheric concentration (≈0.04 %), ISS elevated concentration (0.4 %), and an unexpectedly elevated concentration (1 %) to represent a worst-case scenario. To determine if microbial responses to these CO2 concentrations may be exacerbated in response to the spaceflight environment, microorganisms are being cultured under static conditions and both spaceflight analogue and control conditions using the Rotating Wall Vessel (RWV) bioreactor (Synthecon, Houston, TX). For these studies, we will evaluate the response of Staphylococcus aureus, Streptococcus pneumoniae, and Enterobacter aerogenes, which are opportunistic bacterial pathogens that are of medical significance and have been or are likely to be found aboard human-inhabited spacecraft. Bacteria will be evaluated individually and together as a consortium. The following Specific Aims are be investigated: Aim 1. Characterize the growth, diversity, and pathogenesis-related stress responses of medically significant microorganisms grown under multiple CO2 concentrations under static growth in a liquid medium. Stress responses include acid stress, oxidative stress, and thermal stress. Aim 2. Characterize the growth, diversity, and pathogenesis-related stress responses of medically significant microorganisms grown under multiple CO2 concentrations in both spaceflight analogue and control conditions. Stress responses include acid stress, oxidative stress, and thermal stress.

S G Thornhill↗

Rational Design of Nanoplasmonic Array Geometries for Biosensing

Background: Molecular diagnostics provide early and accurate diagnosis, which is essential for the prevention and treatment of infectious as well as chronic diseases. These tests are designed to detect disease-specific bioanalytes such as nucleic acid (DNA or RNA) or protein (antigens, antibodies) biomarkers. In the context of infectious disease diagnosis, nucleic acid-based detection methods are known to provide more specific and sensitive results. Here, the presence of a unique sequence belonging to the pathogenic genomic material is targeted to identify species, organism, genera and/or antimicrobial resistant gene markers. The majority of the common nucleic acid based diagnostic techniques require amplification (polymerase chain reaction, isothermal amplification etc.) of the pathogenic genetic material prior to detection impacting diagnostic speed, complexity, and cost thereby limiting ease of use. Thus, the development of simplified nucleic acid-based diagnostics that can be even used in resource-poor settings may hugely benefit patients across the globe. Nanopath is a molecular diagnostics company utilizing a solid-state nanosensor to enable sequence-specific detection of target nucleic acids without the need of amplification. These nanostructures enable ultra-sensitive biomarker detection using geometric, feature-dependent properties highly dependent on the local dielectric environment, allowing them to be sensitive to low concentration binding events. This paper describes an application of this approach to provide highly relevant clinical information within a single doctor’s office visit. Intro: The Nanopath team is in collaboration with NASA (National Aeronautics and Space Administration) and NIST (National Institute of Standards and Technology) to push the bounds of the fundamental physics associated with their biosensing platform. The ability of metals to support electromagnetic surface waves gives rise to surface plasmons when optically illuminated. This property, and its strong sensitivity to changes in the local refractive index, allows for the use of metal nanoparticles as ultra-sensitive transducers. In prior work by members of this team, ensembles of randomly oriented nanoparticles (i.e., colloidal nanorods dispersed on chip) were employed for sequence-specific nucleic acid sensing (1-3). While these particle sensors have the advantage of rapid fabrication, they suffer from low sensitivity and quality factor due to the random particle dispersity. In contrast, in this study we employ ordered array nanoparticle ensembles which can be used to improve sensor sensitivity and figure-of-merit. Study Methods Overview: In this talk, we detail the results of sensing experiments and computational simulations to outline a rational design of the structure of these plasmonic nanoparticle arrays for biomolecular sensing. Through simulation and experiment, we iteratively tailor nanostructure dimension to provide high quality signal and large resonance shifts upon modeled nucleic acid binding. In particular, full-wave electromagnetic simulations were conducted using Lumerical photonic simulation software in which periodic boundary conditions were applied in the x- and y- dimensions for each of the nanoplasmonic sensor geometries. To simulate the resonance response to changes in the bulk solution in contact with the sensor surface, the refractive index of the surrounding media was changed appropriately. Nucleic acid hybridization events were modeled using either using spherical structures approximating the relevant radius of genomic material as estimated by polymer models, or as conformal layers with the known refractive indices for nucleic acids. On the basis of initial simulations, nanosensors were fabricated using traditional electron-beam lithography protocols at NIST. To evaluate consensus between simulations and experiments, bulk sensing experiments were carried out in which the resonance peaks were obtained by submerging the sensors in refractive index standards. Key nanosensor characteristics including resonance peak locations, resonance peak shifts as a function of refractive index, and figure of merit (FOM) of extinction curves were examined between the experimental and simulation results prior to proceeding with simulations on additional geometries and more complex solution conditions, and further device fabrication. This iterative process is repeated toward a rational design of nanoplasmonic array geometries for biosensing optimizing response for targeted disease detection. In summary, this study puts forth a methodology for rational design and characterization of regularly spaced nanoparticle arrays for optics-based biosensing. The results of this study will allow for more informed design of nanostructure geometries towards sequence-specific nucleic acid detection. These improved designs have the potential to improve clinical sensitivity and limit-of-detection across disease indication.

sensor↗

Development of Machine Learning-Derived Microbiological and Immune Signatures: Applications in Adaptive Risk Assessment of Infectious Disease During Spaceflight

Infectious diseases represent an urgent risk for spaceflight with consequences ranging from loss in crew performance to crew incapacitation or loss of life should an outbreak occur. The resident environmental microbiome on the International Space Station has been monitored through routine surveillance over almost twenty years, beginning with culture-based microbial detection which has advanced to molecular methods in recent years. This has created a wealth of data that we have begun mining to define the microbial ecology of the ISS. Summarized here is our analysis of data from the historical microbial population defined by culture-based monitoring from the past two decades, organized by their likelihood to cause disease into clinical categories. As expected, many residents of the normal microflora in environments where people work and live were detected. However, some known pathogens were also detected. As the spaceflight environment can predispose humans to infection, crew health records were used to source additional data for the set to uncover clinical relevance. Data mining was performed on crew health records to capture adverse health events that may be related to infectious disease. Machine learning, specifically Random Forest analysis, was used to analyze the microbial and crew health datasets. The symptom categories were not explained by the ranked bacteria, due to lack of sufficient data for some categories and due to poor ranking of the pathogens for others. Poor ranking of the bacteria could be due to the clinical symptoms being linked to other disease-causing factors, such as allergy or viral infection. These findings suggest a lack of relationship between bacteria detected on surfaces in the ISS and historical health events experienced by astronauts.

Kristyn Hoffman↗

The Microbiome of A Tomato Crop Grown Under Different Lighting Regimes on the International Space Station

The VEG-05 experiment presented here investigated the effect of red-rich and blue-rich light recipes in Veggie on the microbiome of the Veggie facility and the plant tissues of a dwarf tomato variety, Solanum lycopersicum cv. Red Robin. For food safety, the plants were screened using culture-based methods for potential human pathogens that may cause infection by consumption of the fruit. The microbiome was investigated using bacterial 16S and fungal ITS sequencing methods to enumerate and identify bacterial and fungal communities on tomato fruit, roots, leaves, rooting substrate, and Veggie facility surfaces grown under blue-rich or red-rich lighting. Comparisons of microbial communities were made between lighting treatments, as well as for flight and ground controls. This analysis determined the core microbiome and microbiological composition for tomato plants grown under a blue-rich or red-rich lighting treatment and microgravity conditions. Culture-based pathogen screening, corroborated by 16S and ITS sequencing, yielded negative results. Bacterial and fungal counts were lower for ground controls than in-flight samples. However, there were no differences in microbial counts between lighting treatments. Regardless of lighting treatment, plant components shared a core microbiome, although some differences were observed in genera between lighting treatments.

Veggie↗

Rift Valley Fever Prediction and Risk Mapping: 2014-2015 Season

Extremes in either direction (+-) of precipitation temperature have significant implications for disease vectors and pathogen emergence and spread Magnitude of ENSO influence on precipitation temperature cannot be currently predicted rely on average history and patterns. Timing of event and emergence disease can be exploited (GAP) in to undertake vector control and preparedness measures. Currently - no risk for ecologically-coupled RVFV activity however we need to be vigilant during the coming fall season due the ongoing buildup of energy in the central Pacific Ocean. Potential for the dual-use of the RVF Monitor system for other VBDs Need to invest in early ground surveillance and the use of rapid field diagnostic capabilities for vector identification and virus isolation.

RVF↗

Surveyor 3: Bacterium isolated from lunar retrieved television camera

Microbial analysis was the first of several studies of the retrieved camera and was performed immediately after the camera was opened. The emphasis of the analysis was placed upon isolating microorganisms that could be potentially pathogenic for man. Every step in the retrieval of the Surveyor 3 television camera was analyzed for possible contamination sources, including camera contact by the astronauts, ingassing in the lunar and command module during the mission or at splashdown, and handling during quarantine, disassembly, and analysis at the Lunar Receiving Laboratory

Mitchell, F. J.↗

Analysis of fungal type isolates taken from a 90-day manned test of an advanced regenerative life support system

Fungal-like cultures isolated before, during, and after the 90-day test from samples of space station simulator (SSS) atmosphere, surfaces, subsystem componets, and crew dermal sites were identified to genus. Out of the original 525 isolates, approximately 80% were classified as bacteria. Laboratory methods (culture media, moisturization, and incubation temperatures) favored the recovery of medically significant bacteria rather than fungi. Therefore, fungal isolates were mostly, nonfastidious types which are ubiquitous in soil and air and commonly contaminate laboratory cultures of pathogens. Predominant isolates were species of Aspergillus, Penicillium, Pullularia, Rhodotorula, and various yeasts. No instances of fungal proliferation were observed; test data reflect the survival of environmental types indigenous to the SSS pretests.

Sofios, M.↗

Microbial ecology measurement system

The sensitivity and potential rapidity of the PIA test that was demonstrated during the feasibility study warranted continuing the effort to examine the possibility of adapting this test to an automated procedure that could be used during manned missions. The effort during this program has optimized the test conditions for two important respiratory pathogens, influenza virus and Mycoplasma pneumoniae, developed a laboratory model automated detection system, and investigated a group antigen concept for virus detection. Preliminary tests on the handling of oropharygeal clinical samples for PIA testing were performed using the adenovirus system. The results obtained indicated that the PIA signal is reduced in positive samples and is increased in negative samples. Treatment with cysteine appeared to reduce nonspecific agglutination in negative samples but did not maintain the signal in positive samples.

Source record↗

Response of tobacco tissue cultures growing in contact with lunar fines.

During the quarantine periods following each Apollo mission to the moon, various biological systems were placed in the presence of lunar material to determine if pathogenic agents were present. Although no detrimental effects resulted, various responses by the several plant systems tested were noted. One such response was the increased pigmentation observed in the callus tissue cultures of tobacco. Further investigations revealed that these tissues grown in the presence of lunar material resulted in as much as a 35% increase in total pigments while differences in fatty acid and sterol concentrations were also noted when compared to the controls. It is believed that these changes brought about by the lunar material can be attributed to a change in the nutritional environment caused by its dissolution.

Weete, J. D.↗

Evaluation of proposed Skylab and SSP soap products.

Four candidate cleansing agents evaluated in terms of potential hazards to crew members included two soaps (Neutrogena bar soap and Olive Leaf Liquid), one nonfoaming surfactant (Miranol JEM), and one laundry detergent (sodium dodecylbenzene sulfonate). None of the four exhibited adverse dermatological effects from skin patch tests or supported growth of potentially pathogenic microorganisms. Aqueous solutions of Neutrogena did support a mold species. Neutrogena and Miranol JEM were used in a simulated Skylab personal hygiene regimen with no adverse effects on skin or skin microflora. Based on our results, each of these agents appear to be a promising candidate material for the use intended.

Durfee, R. L.↗

Storage stability and improvement of intermediate moisture foods, phase 2

Methods for improvement of shelf-life stability of intermediate moisture foods are considered. It was found that vitamin C is the most limiting vitamin from a nutritional standpoint with its rate of destruction increasing with a sub w. Techniques for microbial challenge studies were developed. It was shown that organisms have a higher growth a sub w limit if the IMF is prepared by the adsorption process and long times are needed for challenge studies. Several alternative antimycotic systems were found. It was also found that the vegetative cells of pathogens have a maximum heat resistance in the IMF a sub w range. If glycols are in the formula, the IMF should have as high an a sub w as possible. The reverse is true if lipid oxidation occurs. In addition, to prevent rancidity, antioxidants and a low O2 atmosphere are necessary. The package also must be a good moisture barrier.

Labuza, T. P.↗