Results for “Pathogen” — Page 88

NASA NTRS2017

Micromechanical Devices for Control of Cell-Cell Interaction, and Methods of Use Thereof

The development and function of living tissues depends largely on interactions between cells that can vary in both time and space; however, temporal control of cell-cell interaction is experimentally challenging. By employing a micromachined silicon substrate with moving parts, herein is disclosed the dynamic regulation of cell-cell interactions via direct manipulation of adherent cells with micron-scale precision. The inventive devices and methods allow mechanical control of both tissue composition and spatial organization. The inventive device and methods enable the investigation of dynamic cell-cell interaction in a multitude of applications, such as intercellular communication, spanning embryogenesis, homeostasis, and pathogenic processes.

Bhatia, Sangeeta N.↗

NASA NTRS2017

Microbial Monitoring from the Frontlines to Space: Department of Defense Small Business Innovation Research Technology Aboard the International Space Station

The RAZOR (trademark) EX, a quantitative Polymerase Chain Reaction (qPCR) instrument, is a portable, ruggedized unit that was designed for the Department of Defense (DoD) with its reagent chemistries traceable to a Small Business Innovation Research (SBIR) contract beginning in 2002. The PCR instrument's primary function post 9/11 was to enable frontline soldiers and first responders to detect biological threat agents and bioterrorism activities in remote locations to include field environments. With its success for DoD, the instrument has also been employed by other governmental agencies including Department of Homeland Security (DHS). The RAZOR (Trademark) EX underwent stringent testing by the vendor, as well as through the DoD, and was certified in 2005. In addition, the RAZOR (trademark) EX passed DHS security sponsored Stakeholder Panel on Agent Detection Assays (SPADA) rigorous evaluation in 2011. The identification and quantitation of microbial pathogens is necessary both on the ground as well as during spaceflight to maintain the health of astronauts and to prevent biofouling of equipment. Currently, culture-based monitoring technology has been adequate for short-term spaceflight missions but may not be robust enough to meet the requirements for long-duration missions. During a NASA-sponsored workshop in 2011, it was determined that the more traditional culture-based method should be replaced or supplemented with more robust technologies. NASA scientists began investigating innovative molecular technologies for future space exploration and as a result, PCR was recommended. Shortly after, NASA sponsored market research in 2012 to identify and review current, commercial, cutting edge PCR technologies for potential applicability to spaceflight operations. Scientists identified and extensively evaluated three candidate technologies with the potential to function in microgravity. After a thorough voice-of-the-customer trade study and extensive functional and safety evaluations, the RAZOR (trademark) EX PCR instrument(Bio-Fire Defense, Salt Lake City, UT) was selected as the most promising current technology for spaceflight monitoring applications.

Cherie M Oubre↗

NASA NTRS2017

Expansion of Microbial Monitoring Capabilities on the International Space Station (ISS)

Microbial monitoring is one of the tools that the National Aeronautics and Space Administration (NASA) uses on the International Space Station (ISS) to help maintain crew health and safety. In combination with regular housekeeping and disinfection when needed, microbial monitoring provides important information to the crew about the quality of the environment. Rotation of astronauts, equipment, and cargo on the ISS can affect the microbial load in the air, surfaces, and water. The current ISS microbial monitoring methods are focused on culture-based enumeration during flight and require a significant amount of crew time as well as long incubation periods of up to 5 days there by proliferating potential pathogens. In addition, the samples require return to Earth for complete identification of the microorganisms cultivated. Although the current approach assess the quality of the ISS environment, molecular technology offers faster turn-around of information particularly beneficial in an off-nominal situation. In 2011, subject matter experts from industry and academia recommended implementation of molecular-based technologies such as quantitative real-time polymerase chain reaction (qPCR) for evaluation to replace current, culture-based technologies. The RAZOR EX (BioFire Defense, Inc, Salt Lake City, UT) a ruggedized, compact, COTS (commercial off the shelf) qPCR instrument was tested, evaluated and selected in the 2 X 2015 JSC rapid flight hardware demonstration initiative as part of the Water Monitoring Suite. RAZOR EX was launched to ISS on SpaceX-9 in July 2016 to evaluate the precision and accuracy of the hardware by testing various concentrations of DNA in microgravity compared to ground controls. Flight testing was completed between September 2016 and March 2017. Data presented will detail the hardware performance of flight testing results compared to ground controls. Future goals include additional operational ground-based testing and assay development to determine if this technology can meet spaceflight microbial monitoring requirements.

Khodadad, Christina L.↗

NASA NTRS2017

Future Food Production System Development Pulling from Space Biology Crop Growth Testing in Veggie

Preliminary crop testing using Veggie indicates the environmental conditions provided by the ISS are generally suitable for food crop production. When plant samples were returned to Earth for analysis, their levels of nutrients were comparable to Earth-grown ground controls. Veggie-grown produce food safety microbiology analysis indicated that space-grown crops are safe to consume. Produce sanitizing wipes were used on-orbit to further reduce risk of foodborne illness. Validation growth tests indicated abiotic challenges of insufficient or excess fluid delivery, potentially reduced air flow leading to excess water, elevated CO2 leading to physiological responses, and microorganisms that became opportunistic pathogens. As NASA works to develop future space food production, several areas of research to define these systems pull from the Veggie technology validation tests. Research into effective, reusable water delivery and water recovery methods for future food production systems arises from abiotic challenges observed. Additionally, impacts of elevated CO2 and refinement of fertilizer and light recipes for crops needs to be assessed. Biotic pulls include methods or technologies to effectively sanitize produce with few consumables and low inputs; work to understand the phytomicrobiome and potentially use it to protect crops or enhance growth; selection of crops with high harvest index and desirable flavors for supplemental nutrition; crops that provide psychosocial benefits, and custom space crop development. Planning for future food production in a deep space gateway or a deep space transit vehicle requires methods of handling and storing seeds, and ensuring space seeds are free of contaminants and long-lived. Space food production systems may require mechanization and autonomous operation, with preliminary testing initiated to identify operations and capabilities that are candidates for automation. Food production design is also pulling from Veggie logistics lessons, as we learn about growing at different scales and move toward developing systems that require less launch mass. Veggie will be used as a test bed for novel food production technologies. Veggie is a relatively simple precursor food production system but the knowledge gained from space biology validation tests in Veggie will have far reaching repercussions on future exploration food production.

Veggie↗

NASA NTRS2017

Future Food Production System Development Pulling From Space Biology Crop Growth Testing in Veggie

Preliminary crop testing using Veggie indicates the environmental conditions provided by the ISS are generally suitable for food crop production. When plant samples were returned to Earth for analysis, their levels of nutrients were comparable to Earth-grown ground controls. Veggie-grown produce food safety microbiology analysis indicated that space-grown crops are safe to consume. Produce sanitizing wipes were used on-orbit to further reduce risk of foodborne illness. Validation growth tests indicated abiotic challenges of insufficient or excess fluid delivery, potentially reduced air flow leading to excess water, elevated CO2 leading to physiological responses, and microorganisms that became opportunistic pathogens. As NASA works to develop future space food production, several areas of research to define these systems pull from the Veggie technology validation tests. Research into effective, reusable water delivery and water recovery methods for future food production systems arises from abiotic challenges observed. Additionally, impacts of elevated CO2 and refinement of fertilizer and light recipes for crops needs to be assessed. Biotic pulls include methods or technologies to effectively sanitize produce with few consumables and low inputs; work to understand the phytomicrobiome and potentially use it to protect crops or enhance growth; selection of crops with high harvest index and desirable flavors for supplemental nutrition; crops that provide psychosocial benefits, and custom space crop development. Planning for future food production in a deep space gateway or a deep space transit vehicle requires methods of handling and storing seeds, and ensuring space seeds are free of contaminants and long-lived. Space food production systems may require mechanization and autonomous operation, with preliminary testing initiated to identify operations and capabilities that are candidates for automation. Food production design is also pulling from Veggie logistics lessons, as we learn about growing at different scales and move toward developing systems that require less launch mass. Veggie will be used as a test bed for novel food production technologies. Veggie is a relatively simple precursor food production system but the knowledge gained from space biology validation tests in Veggie will have far reaching repercussions on future exploration food production. This work is supported by NASA.

Plants↗

NASA NTRS2018

Baseline Microbial Assessment of Fresh Produce

Currently no standards or requirements exist for microbial food safety for space grown produce (fresh plant foods). Without standards it is difficult to assess produce handling and sanitization options for the ISS and future exploration missions. We are conducting a literature review of microbial levels on fresh food and then carrying out measurements (microbial counts) of grocery store purchased and controlled environment-grown crops. Testing will include lettuce, mizuna, cherry tomato, pepper, and radish, all candidate crops for pick-and-eat testing on ISS and near term exploration missions. Growth chamber conditions will be set to mimic an ISS or spacecraft environment. Assays will include specific pathogens (Enterobacteriacea, Salmonella sp., and Aspergillus flavus) and total culturable microorganisms using aerobic plate counts, and total yeast and mold counts. Analyses will follow the FDA Bacteriological Analytical Manual methods. The goal of the project is to establish a baseline for expected microbial levels found on fresh plant foods that might be grown on ISS and near term missions, and develop risk assessment and microbial safety recommendations for these types of fresh foods.

Hummerick, M. E.↗

NASA NTRS2018

Antimicrobials for Water Systems in Manned Spaceflight - Past, Present, and Future Applications and Challenges

The use of antimicrobials to control microbiological growth in manned spaceflight water-based systems has and will continue to have a unique set of challenges and needs. The challenges are varied, and include antimicrobial effectiveness, crew health and safety, materials compatibility, optimal system functionality, antimicrobial shelf life, means to monitor antimicrobial concentration, and means to re-introduce biocides periodically in the case of depletion. Needs vary from application to application, and include control of pathogens for crew health, control of biofilm formation for optimal system functionality, inhibition and prevention of microbiologically influenced corrosion, optimization of wetted metallic material life, and general living quarter and consumable aesthetics with respect to odor and taste. This paper outlines and discusses the various antimicrobials used in prior and current manned spaceflight water-based applications with focus on pros, cons and lessons learned. Design factors such as minimum inhibitory concentration, minimum lethal concentration, required circulated concentrations, materials selection, means to introduce, means to monitor real-time, and concentration maintenance are discussed. The challenges associated with longer term missions, as well as long-term system dormancy as envisioned for exploration missions, lunar habitats, and a manned Mars mission are outlined with respect to anticipated needs and potential design solutions.

potable water↗

NASA NTRS2018

Mobile/Modular BSL-4 Containment Facilities Integrated into a Curation Receiving Laboratory for Restricted Earth Return Missions

NASA robotic sample return missions designated Category V Restricted Earth Return by the NASA Planetary Protection (PP) Office require sample containment and biohazard testing upon return to Earth. Since the 1960s, sample containment from an unknown extraterrestrial biohazard have been related to the highest containment standards and protocols known to modern science. Today, this is Biosafety Level (BSL) 4 containment. In the U.S., the Biosafety in Microbiological and Biomedical Laboratories publication authored by the U.S. Department of Health and Human Services (HHS): Public Health Service, Centers for Disease Control and Prevention, and the National Institutes of Health houses the primary recommendations, standards, and design requirements for all BSL labs. Past mission concept studies for constructing a NASA Curation Receiving Laboratory with an integrated BSL-4 quarantine and biohazard testing facility have been estimated in the hundreds of millions of dollars (USD). As an alternative option, we have conducted a trade study for constructing a mobile and/or modular sample containment laboratory that would meet all BSL-4 and planetary protection standards and protocols at a fraction of the cost. Mobile and modular BSL-2 and 3 facilities have been successfully constructed and deployed world-wide for government testing of pathogens and pharmaceutical production. Our study showed that a modular BSL-4 construction could result in ~ 90% cost reduction when compared to traditional BSL-4 construction methods without compromising the preservation of the samples or Earth. For the design/construction requirements of a mobile/modular BSL-4 containment, we used the established HHS document standards and protocols for manipulation of agents in Class III Biosafety Cabinets (BSC; i.e., negative pressure gloveboxes) that are currently followed in operational BSL-4 facilities in the U.S.

Calaway, Michael J.↗

NASA NTRS2014

Innate Immune Responses of Drosophila Melanogaster are Altered by Spaceflight

Alterations and impairment of immune responses in humans present a health risk for space exploration missions. The molecular mechanisms under pinning innate immune defense can be confounded by the complexity of the acquired immune system of humans. Drosophila (fruit fly) innate immunity is simpler, and shares many similarities with human innate immunity at the level of molecular and genetic pathways. The goals of this study were to elucidate fundamental immune processes in Drosophila affected by spaceflight and to measure host-pathogen responses post-flight. Five containers, each containing ten female and five male fruit flies, were housed and bred on the space shuttle (average orbit altitude of330.35 km) for 12 days and 18.5 hours. A new generation of flies was reared in microgravity. In larvae, the immune system was examined by analyzing plasmatocyte number and activity in culture. In adults, the induced immune responses were analyzed by bacterial clearance and quantitative real-time polymerase chain reaction (qPCR) of selected genes following infection with E. coli. The RNA levels of relevant immune pathway genes were determined in both larvae and adults by microarray analysis. The ability of larval plasmatocytes to phagocytose E. coli in culture was attenuated following spaceflight, and in parallel, the expression of genes involved in cell maturation was down regulated. In addition, the level of constitutive expression of pattern recognition receptors and opsonins that specifically recognize bacteria, and of lysozymes, antimicrobial peptide (AMP) pathway and immune stress genes, hallmarks of humoral immunity, were also reduced in larvae. In adults, the efficiency of bacterial clearance measured in vivo following a systemic infection with E. coli post-flight, remained robust. We show that spaceflight altered both cellular and humoral immune responses in Drosophila and that the disruption occurs at multiple interacting pathways.

Marcu, Oana↗

NASA NTRS2018

Stress-Induced Heat Shock Protein 40 and Immune Function in Altered Gravity

In space, astronauts are more susceptible to pathogens, viral reactivation and immunosuppression, which poses limits to their health and the mission. Interestingly, during space flight, stress-inducible heat shock proteins (HSP) are robustly induced, and the overexpression of HSPs have been implicated in immune dysregulation, therefore HSPs may be critically involved in regulating immune homeostasis. HSP40/DNAJ1 plays a major role in proper protein translation and folding. Its loss of function has been implicated in susceptibility to microbial infection, while its overexpression has been implicated in autoimmunity, collectively suggesting its complicated, but necessary, role in maintaining immunological function. To determine the role of HSP40 during stress-induced altered gravity conditions, wild-type and Hsp40 mutant Drosophila melanogaster were exposed to ground-based chronic hypergravity conditions, followed by quantitative PCR (qPCR) analysis of immune gene expression. In addition, larval hemocytes were collected to determine the functional output in response to E. coli bioparticle phagocytosis. Preliminary data indicates a required role for Hsp40 in strengthening immune function during stress-induced spaceflight in flies. In short, a critical need to evaluate the relationship between HSPs and immune suppression during space flight is necessary. Since space travel may become available to the general public in the not too distant future, and for the possibility of long-term space missions, a more comprehensive evaluation of the molecules responsible for immune dysfunction observed during space flight is required.

hypergravity↗

NASA NTRS2014

E. coli AntiMicrobial Satellite (EcAMSat)

The E. coli AntiMicrobial Satellite(EcAMSat) mission will investigate space microgravity affects on the antibiotic resistance of E. coli, a bacterial pathogen responsible for urinary tract infection in humans and animals. EcAMSat is being developed through a partnership between NASA Ames Research Center and the Stanford University School of Medicine. Scientists believe that the results of this experiment could help design effective countermeasures to protect astronauts health during long duration human space missions.

e. Coli↗

NASA NTRS2019

Global Disease Outbreaks Associated with the 2015-2016 El Niño Event

Interannual climate variability patterns associated with the El Niño-Southern Oscillation phenomenon result in climate and environmental anomaly conditions in specific regions worldwide that directly favor outbreaks and/or amplification of variety of diseases of public health concern including chikungunya, hantavirus, Rift Valley fever, cholera, plague, and Zika. We analyzed patterns of some disease outbreaks during the strong 2015–2016 El Niño event in relation to climate anomalies derived from satellite measurements. Disease outbreaks in multiple El Niño-connected regions worldwide (including Southeast Asia, Tanzania, western US, and Brazil) followed shifts in rainfall, temperature, and vegetation in which both drought and flooding occurred in excess (14–81% precipitation departures from normal). These shifts favored ecological conditions appropriate for pathogens and their vectors to emerge and propagate clusters of diseases activity in these regions. Our analysis indicates that intensity of disease activity in some ENSO-teleconnected regions were approximately 2.5–28% higher during years with El Niño events than those without. Plague in Colorado and New Mexico as well as cholera in Tanzania were significantly associated with above normal rainfall (p < 0.05); while dengue in Brazil and southeast Asia were significantly associated with above normal land surface temperature (p < 0.05). Routine and ongoing global satellite monitoring of key climate variable anomalies calibrated to specific regions could identify regions at risk for emergence and propagation of disease vectors. Such information can provide sufficient lead-time for outbreak prevention and potentially reduce the burden and spread of ecologically coupled diseases.

El Nino↗

NASA NTRS2018

Heat Shock Protein 40 and Immune Function in Altered Gravity

In space, astronauts are more susceptible to pathogens, viral reactivation and immunosuppression, which poses limits to their health and the mission. Interestingly, during space flight, stress-inducible heat shock proteins (HSP) are robustly induced, and the overexpression of HSPs have been implicated in immune dysregulation, therefore HSPs may be critically involved in regulating immune homeostasis. HSP40/DNAJ1 plays a major role in proper protein translation and folding. Its loss of function has been implicated in susceptibility to microbial infection, while its overexpression has been implicated in autoimmunity, collectively suggesting its complicated, but necessary, role in maintaining immunological function. To determine the role of HSP40 during stress-induced altered gravity conditions, wild-type and Hsp40 mutant Drosophila melanogaster were exposed to ground-based chronic hypergravity conditions, followed by quantitative PCR (qPCR) analysis of immune gene expression. In addition, larval hemocytes were collected to determine the functional output in response to E. coli bioparticle phagocytosis. Preliminary data indicates a required role for Hsp40 in strengthening immune function during stress-induced spaceflight in flies. In short, a critical need to evaluate the relationship between HSPs and immune suppression during space flight is necessary. Since space travel may become available to the general public in the not too distant future, and for the possibility of long-term space missions, a more comprehensive evaluation of the molecules responsible for immune dysfunction observed during space flight is required.

hypergravity↗

NASA NTRS2019

Survival of E. Coli in the Rhizosphere and Phyllosphere of Leafy Greens Grown in Controlled Environment Chambers Under International Space Station Conditions

NASA's mission for manned long- duration space exploration drives the research for crop selection to provide a nutritious and safe supplement to an astronaut's diet. Understanding plant growth, health, and the associated microbial communities in closed environments will be critical to the success of this mission. Cultivation of crops in closed controlled environment agricultural systems may limit microbial colonization and reduce diversity of the microbial communities. Furthermore, practices like seed and growth medium sanitization may impact microbial communities in the mature plant and the capacity to limit the growth of food borne pathogens through competition.

Hummerick, Mary E.↗

NASA NTRS2019

Investigation of Biofilm Formation and Control for Spacecraft-An Early Literature Review

Bacterial biofilms are an important and often problematic aspect of life on earth and inspace. Biofilms of opportunistic pathogenic bacteria can lead to severe and costlycontamination problems that directly affect human health and long-term mission planning.Microbial contamination on board the International Space Station (ISS) continues to posemission risks, both to crew health and hardware reliability. In order to optimize the design offuture space exploration vehicles, a thorough understanding of biofilm formation and controltechnologies is needed to control the habitat's microbial environment. This paper provides aliterature review on microbial behavior, biofilm formation in spacecraft or simulatedspacecraft environments, and the state of the art of biofilm prevention mechanisms

Diaz, Angie M.↗

NASA NTRS2019

The Effects of Low-Shear Simulated Microgravity on Soil Bacterium Pantoea Agglomerans

As NASA's effort to establish a permanent residence in space continues, research on the effects of microgravity onbiological microorganisms is vital to protect or promote the health of plants and their astronaut counterparts. The purpose of this study is to determine the effects of microgravity on Pantoea agglomerans (P. agglomerans), using an analog microgravity simulator; the Rotary Cell Culture System (RCCS) developed at Johnson Space Center (JSC) in Houston, TX. P. agglomerans has been shown to be a plant growth promoter (PGPR) in ground based studies, but has also been shown to be a pathogen in both plants and immunocompromised patients. In this study, we will determine changes in the growth rate and antibiotic susceptibility of P. agglomerans when exposed to simulated microgravity.

HARV↗

NASA NTRS2019

Biological CubeSats: What Have We Learned so Far and What Is Next?

Since Apollo 17 in 1972, NASA has sent no humans or other biological organisms outside of Earth's protective magnetosphere. Recently, NASA has set its sights on human exploration in deep space, with an ambitous plan to put astronauts back on the Moon by 2024 and to eventually land human missions on Mars. Such missions will require significant countermeasures, likely both technological and biomedical, to protect biology from chronic radiation exposure. CubeSats can inform these countermeasures by querying relevant space environments with model organisms.NASA has launched five biological CubeSat missions into low-Earth orbit (LEO). GeneSat-1 was launched in 2006 to study gene expression and increase our knowledge of how spaceflight affects microbes. Similar life-support technologies were then used in PharmaSat and O/OREOS, which launched in 2009 and 2010, respectively. PharmaSat contained optical systems to examine how yeast cells responded to an antifungal treatment. One of O/OREOS payloads, SESLO (Space Environment Survivability of Living Organisms), housed dormant microorganisms, which were rehydrated on orbit to track alterations to growth and metabolism induced by microgravity and radiation. In 2014, NASA launched SporeSat to study the mechanisms of plant cell gravity sensing using lab-on-a-chip devices. Most recently, in 2017, NASA launched EcAMSat (E. coli AntiMicrobial Satellite), which investigated the effects of microgravity on antibiotic resistance of a pathogenic bacterium. Each one of these missions increased our understanding of the biological effects of spaceflight in LEO, while refining technologies and imparting valuable lessons to the next generation of CubeSats.CubeSats housing translational biological models are therefore ideal for defining the hazards of deep space travel, as they can provide critical data over relevant durations. BioSentinel, a next-generation deep-space CubeSat, is planned to launch as a secondary payload on Artemis 1 in 2020. BioSentinel will study the DNA damage response to deep space radiation in yeast.

Santa Maria, Sergio R.↗

NASA NTRS2016

On-orbit RNA Purification and qRT-PCR Capabilities of the WetLab-2 System

NASA Ames Research Center’s WetLab-2 system brings new capabilities to the ISS for researchers. The system can lyse cells and extract RNA on-orbit from different sample types ranging from microbial cultures to animal tissues. Our purification method has the advantage of using non-toxic chemicals and does not require alcohols or other organics. The purified RNA can then either be stabilized for return to Earth or can be used to conduct on-orbit quantitative Reverse Transcriptase PCR (qRT-PCR) analysis without the need for sample return. qRT-PCR reactions are performed by dispensing the RNA into reaction tubes that contain all lyophilized reagents needed to perform the analysis. The system uses a Cepheid® SmartCycler that allows for multiplexing of assays, this can be used to normalize for RNA concentration and integrity and to study multiple genes of interest in each tube. There are a total of 16 independent PCR modules each capable of detecting up to four fluorescent channels. The WetLab- 2 system can downlink data from the ISS to the ground after a completed run and uplink new thermal cycling programs. The ability to purify and stabilize RNA on-orbit can eliminate the confounding effects of reentry stresses and shock acting on live cells and organisms or the concern of RNA degradation of some samples. It also has the benefit of minimizing the needed downmass. Conducting qRT-PCR and generating results on-orbit is also an important step towards utilizing the ISS as a National Laboratory facility. Specifically, the ability to get on-orbit data will provide investigators with the opportunity to adjust experimental parameters in real time without the need for sample return and re-flight. On orbit gene expression analysis can also provide benchmarking prior to sample return. The system can also be used for analysis of air, surface, water, and clinical samples to monitor environmental pathogens and crew health. The validation flight of the WetLab-2 system using E. coli bacteria and mouse liver is scheduled to launch on SpaceX-8 this spring. Pending operations, the preliminary results from the validation flight will be presented. To support the needs of future researchers, we are adapting our system to purify RNA from two additional sample types: fibrous tissue such as muscle and mammalian adherent cells grown on alginate beads. Progress of this work will also be presented. The WetLab-2 Project is supported by the Research Integration Office in the ISS Program.

Parra, Macarena↗