scholarly journals Design and Validation of a Device for Mitigating Fluid Microgravity Effects in Biological Research in Canister Spaceflight Hardware

2021 ◽  
Vol 2 ◽  
Author(s):  
Wayne L. Nicholson ◽  
Patricia Fajardo-Cavazos ◽  
Caleb Turner ◽  
Taylor M. Currie ◽  
Geoffrey Gregory ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Space Station ◽  
Petri Dish ◽  
Microgravity Environment ◽  
Direct Result ◽  
Ground Control ◽  
Biological Research ◽  
Transcriptomic Response ◽  
Liquid Cultures ◽  
Control Samples

The major factor influencing the behavior of microbes growing in liquids in space is microgravity. We recently measured the transcriptomic response of the Gram-positive bacterium Bacillus subtilis to the microgravity environment inside the International Space Station (ISS) in spaceflight hardware called Biological Research in Canisters-Petri Dish Fixation Units (BRIC-PDFUs). In two separate experiments in the ISS, dubbed BRIC-21 and BRIC-23, we grew multiple replicates of the same B. subtilis strain in the same hardware, growth medium, and temperature with matching ground control samples (npj Micrograv. 5:1.2019, doi: 10.1038/s41526-018-0061-0). In both experiments we observed similar responses of the transcriptome to spaceflight. However, we also noted that the liquid cultures assumed a different configuration in microgravity (a toroidal shape) compared with the ground control samples (a flat disc shape), leading us to question whether the transcriptome differences we observed were a direct result of microgravity, or a secondary result of the different liquid geometries of the samples affecting, for example, oxygen availability. To mitigate the influence of microgravity on liquid geometry in BRIC canisters, we have designed an insert to replace the standard 60-mm Petri dish in BRIC-PDFU or BRIC-LED sample compartments. In this design, liquid cultures are expected to assume a more disk-like configuration regardless of gravity or its absence. We have: (i) constructed a prototype device by 3D printing; (ii) evaluated different starting materials, treatments, and coatings for their wettability (i.e., hydrophilicity) using contact angle measurements; (iii) confirmed that the device performs as designed by drop-tower testing and; (iv) performed material biocompatibility studies using liquid cultures of Bacillus subtilis and Staphylococcus aureus bacteria. Future microgravity testing of the device in the ISS is planned.

Establishing Standard Protocols for Bacterial Culture in Biological Research in Canisters (BRIC) Hardware

2020 ◽  
Vol 4 (2) ◽  
pp. 58-69 ◽  
Author(s):  
Patricia Fajardo-Cavazos ◽  
Wayne L. Nicholson
Keyword(s):  
Gene Expression ◽  
Bacterial Culture ◽  
Culture Conditions ◽  
Media Culture ◽  
Ground Control ◽  
Biological Research ◽  
Confounding Factors ◽  
Cross Experiment ◽  
Spaceflight Experiments ◽  
Control Samples

AbstractThe NASA GeneLab Data System (GLDS) was recently developed to facilitate cross-experiment comparisons in order to understand the response of microorganisms to the human spaceflight environment. However, prior spaceflight experiments have been conducted using a wide variety of different hardware, media, culture conditions, and procedures. Such confounding factors could potentially mask true differences in gene expression between spaceflight and ground control samples. In an attempt to mitigate such confounding factors, we describe here the development of a standardized set of hardware, media, and protocols for liquid cultivation of microbes in Biological Research in Canisters (BRIC) spaceflight hardware, using the model bacteria Bacillus subtilis strain 168 and Staphylococcus aureus strain UAMS-1 as examples.

scholarly journals Nanosatellites for Biology in Space: In Situ Measurement of Bacillus subtilis Spore Germination and Growth after 6 Months in Low Earth Orbit on the O/OREOS Mission

Life ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 1 ◽  
Author(s):  
Wayne L. Nicholson ◽  
Antonio J. Ricco
Keyword(s):  
Bacillus Subtilis ◽  
Optical Density ◽  
Spore Germination ◽  
Space Environment ◽  
Ground Control ◽  
Earth Orbit ◽  
Growth Responses ◽  
Germination And Growth ◽  
Control Samples

We report here complete 6-month results from the orbiting Space Environment Survivability of Living Organisms (SESLO) experiment. The world’s first and only long-duration live-biology cubesat experiment, SESLO was executed by one of two 10-cm cube-format payloads aboard the 5.5-kg O/OREOS (Organism/Organic Exposure to Orbital Stresses) free-flying nanosatellite, which launched to a 72°-inclination, 650-km Earth orbit in 2010. The SESLO experiment measured the long-term survival, germination, metabolic, and growth responses of Bacillus subtilis spores exposed to microgravity and ionizing radiation including heavy-ion bombardment. A pair of radiation dosimeters (RadFETs, i.e., radiation-sensitive field-effect transistors) within the SESLO payload provided an in-situ dose rate estimate of 6–7.6 mGy/day throughout the mission. Microwells containing samples of dried spores of a wild-type B. subtilis strain and a radiation-sensitive mutant deficient in Non-Homologoous End Joining (NHEJ) were rehydrated after 14, 91, and 181 days in space with nutrient medium containing with the redox dye alamarBlue (aB), which changes color upon reaction with cellular metabolites. Three-color transmitted light intensity measurements of all microwells were telemetered to Earth within days of each 24-hour growth experiment. At 14 and 91 days, spaceflight samples germinated, grew, and metabolized significantly more slowly than matching ground-control samples, as measured both by aB reduction and optical density changes; these rate differences notwithstanding, the final optical density attained was the same in both flight and ground samples. After 181 days in space, spore germination and growth appeared hindered and abnormal. We attribute the differences not to an effect of the space environment per se, as both spaceflight and ground-control samples exhibited the same behavior, but to a pair of ~15-day thermal excursions, after the 91-day measurement and before the 181-day experiment, that peaked above 46 °C in the SESLO payload. Because the payload hardware operated nominally at 181 days, the growth issues point to heat damage, most likely to component(s) of the growth medium (RPMI 1640 containing aB) or to biocompatibility issues caused by heat-accelerated outgassing or leaching of harmful compounds from components of the SESLO hardware and electronics.

scholarly journals Methods for On-Orbit Germination of Arabidopsis thaliana for Proteomic Analysis

2020 ◽  
Vol 4 (2) ◽  
pp. 20-27
Author(s):  
Sarahann Hutchinson ◽  
Proma Basu ◽  
Sarah E. Wyatt ◽  
Darron R. Luesse
Keyword(s):  
Proteomic Analysis ◽  
Large Scale ◽  
Space Station ◽  
Petri Dish ◽  
Biological Research ◽  
Original Research ◽  
Temperature Growth ◽  
Proteomic Approach ◽  
Crowding Stress ◽  
The Impact

AbstractLarge-scale omics approaches make excellent choices for research aboard the International Space Station (ISS) because they provide large amounts of data that can be continually mined even after the original research has been completed. A proteomic approach can provide information about which proteins are produced, degraded, or post-translationally modified, potentially shedding light on cellular strategies that cannot be discerned from transcriptomic data. To collect sufficient tissue from a Biological Research In Canisters (BRIC)-grown experiment on the ISS for proteomic analysis, several modifications were made to existing protocols. Approximately 800–1000 seeds were housed in each Petri Dish Fixation Units (PDFU). These were germinated up to 120 h after planting by transferring the BRIC from cold stasis to room temperature. Growth continued for only 72 h after germination to allow sufficient tissue for extraction, and to minimize the impact of ethylene and crowding stress. Seedlings were then exposed to RNAlater®. Results indicate that RNAlater® - treated Arabidopsis seedlings yield an equal amount of protein to those flash-frozen in liquid nitrogen.

scholarly journals Space Flight Effects on Antioxidant Molecules in Dry Tardigrades: The TARDIKISS Experiment

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Angela Maria Rizzo ◽  
Tiziana Altiero ◽  
Paola Antonia Corsetto ◽  
Gigliola Montorfano ◽  
Roberto Guidetti ◽  
...  
Keyword(s):  
Space Shuttle ◽  
Space Station ◽  
Total Content ◽  
Thiobarbituric Acid ◽  
Space Experiment ◽  
Microgravity Environment ◽  
Dna Integrity ◽  
Ros Scavenging ◽  
And Control ◽  
Control Samples

The TARDIKISS (Tardigrades in Space) experiment was part of the Biokon in Space (BIOKIS) payload, a set of multidisciplinary experiments performed during the DAMA (Dark Matter) mission organized by Italian Space Agency and Italian Air Force in 2011. This mission supported the execution of experiments in short duration (16 days) taking the advantage of the microgravity environment on board of the Space Shuttle Endeavour (its last mission STS-134) docked to the International Space Station. TARDIKISS was composed of three sample sets: one flight sample and two ground control samples. These samples provided the biological material used to test as space stressors, including microgravity, affected animal survivability, life cycle, DNA integrity, and pathways of molecules working as antioxidants. In this paper we compared the molecular pathways of some antioxidant molecules, thiobarbituric acid reactive substances, and fatty acid composition between flight and control samples in two tardigrade species, namely,Paramacrobiotus richtersiandRamazzottius oberhaeuseri. In both species, the activities of ROS scavenging enzymes, the total content of glutathione, and the fatty acids composition between flight and control samples showed few significant differences. TARDIKISS experiment, together with a previous space experiment (TARSE), further confirms that both desiccated and hydrated tardigrades represent useful animal tool for space research.

scholarly journals Exogenous pigments shield microorganisms from spaceflight-induced changes

2021 ◽  
Author(s):  
Sunanda Sharma ◽  
Rachel Soo Hoo Smith ◽  
Nicolas A Lee ◽  
Sara Laura Wilson ◽  
Miana M Smith ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Space Station ◽  
Carotenoid Pigments ◽  
Liquid Cultures ◽  
International Space ◽  
Induced Stress ◽  
Solid Media ◽  
Induced Changes ◽  
Distinct Morphology ◽  
Morphology Changes

Research has indicated that pigments commonly produced by microorganisms may be protective against the environmental stresses inherent to spaceflight. However, few studies have directly tested the protective capabilities of microbial pigments applied externally as shielding materials. In this study, liquid cultures of Bacillus subtilis were shielded by various pigment solutions, and solid media cultures of Bacillus subtilis were co-inoculated with the highly pigmented microorganisms Aspergillus niger and Neurospora crassa. These experiments were conducted in a compact, automated payload aboard the International Space Station (ISS) interior for 30 days. Post-flight phenotypic analyses of liquid cultures showed that solutions of carotenoid pigments were effective at minimizing detrimental effects of spaceflight. Elevated growth rate was observed for solid cultures, and distinct morphology changes were identified in both liquid and solid samples and quantified as markers of spaceflight-induced stress. These findings collectively progress our understanding of microbial pigments for the development of space-related applications.

scholarly journals Growth Inhibition of Cocoa Pod Rot Fungus Phytophthora palmivora byPseudomonas fluorescence and Bacillus subtilis bacteria

2013 ◽  
Vol 29 (2) ◽  
Author(s):  
Sakti Widyanta Pratama ◽  
Sri Sukamto ◽  
Lis Nur Asyiah ◽  
Yeni Vida Ervina
Keyword(s):  
Bacillus Subtilis ◽  
Fungal Growth ◽  
Petri Dish ◽  
Phytophthora Palmivora ◽  
Fungal Treatment ◽  
Pure Cultures ◽  
Black Pod ◽  
Black Pod Disease ◽  
First Time ◽  
Important Disease

Black pod disease caused by Phytophthora palmivorafungus is one of the important diseases on cocoa crop. Pod rot is the most important disease because it may cause loss of cocoa pod. Until now, the fungal pathogen of cocoa black pod disease is still a crucial problem and there is no fungicide that is really effective against the disease. One alternative to control the cocoa black pod disease is by using biological agents as biofungicide, including utilizing Pseudomonas fluorescenceand Bacillus subtilis bacteria. The research was done by isolation of P. palmivora from infected pods of Kaliwining Experimental Station to obtain pure cultures of fungus and by multiplication of P. fluorescence and B. subtilis. Antagonist test was performed by inoculating P. palmivora into a petri dish in a distance of 3 cm from the edge. P. fluorescenceand B. Subtilis were inoculated into petridishes in three days after the fungal treatment. Control was inoculated with isolate of P. palmivora only. Fungal growth was measured everyday by measuring radius of fungal colonies first time 24 hours after inoculation. Growth of Phytophthora palmivora in the two treatmens were used to calculate the percentage of inhibition. The results of this study indicated that P. fluorescence and B. subtiliswere able to inhibit fungal growth of P. palmivora. Both bacterial antagonists had the same effectiveness in inhibiting the growth of P. palmivora fungus based on the percentage of inhibition and effectiveness criteria. Based on the results of translucent zones indicated that B. subtiliswas more powerfull in inhibiting growth of P. Palmivora compared to P. fluorescence. Key words: Black pod disease of cocoa, biological control, Phytophthora palmivora, Pseudomonas fluorescence, Bacillus subtilis

scholarly journals Comparative Transcriptomic Signature of the Simulated Microgravity Response in Caenorhabditis elegans

2019 ◽  
Author(s):  
İrem Çelen ◽  
Aroshan Jayasinghe ◽  
Jung H. Doh ◽  
Chandran R. Sabanayagam
Keyword(s):  
Caenorhabditis Elegans ◽  
Simulated Microgravity ◽  
Space Station ◽  
Integrative Approach ◽  
Biological Processes ◽  
Rna Seq ◽  
Transcriptomic Response ◽  
Transcriptomic Signature ◽  
Ground Conditions ◽  
The Impact

AbstractBackgroundGiven the growing interest in human exploration of space, it is crucial to identify the effect of space conditions on biological processes. The International Space Station (ISS) greatly helps researchers determine these effects. However, the impact of the ISS-introduced potential confounders (e.g., the combination of radiation and microgravity exposures) on the biological processes are often neglected, and separate investigations are needed to uncover the impact of individual conditions.ResultsHere, we analyze the transcriptomic response of Caenorhabditis elegans to simulated microgravity and observe the maintained transcriptomic response after return to ground conditions for four, eight, and twelve days. Through the integration of our data with those in NASA GeneLab, we identify the gravitome, which we define as microgravity-responsive transcriptomic signatures. We show that 75% of the simulated microgravity-induced changes on gene expression persist after return to ground conditions for four days while most of these changes are reverted after twelve days return to ground conditions. Our results from integrative RNA-seq and mass spectrometry analyses suggest that simulated microgravity affects longevity regulating insulin/IGF-1 and sphingolipid signaling pathways.ConclusionsOur results address the sole impact of simulated microgravity on transcriptome by controlling for the other space-introduced conditions and utilizing RNA-seq. Using an integrative approach, we identify a conserved transcriptomic signature to microgravity and its sustained impact after return to the ground. Moreover, we present the effect of simulated microgravity on distinct ceramide profiles. Overall, this work can provide insights into the sole effect of microgravity on biological systems.

scholarly journals Design of a Small-Scale Experimental Model of the International Space Station Crew Quarters for a PIV Flow Field Study

2019 ◽  
Vol 111 ◽  
pp. 01045
Author(s):  
Matei-Razvan Georgescu ◽  
Ilinca Nastase ◽  
Amina Meslem ◽  
Mihnea Sandu ◽  
Florin Bode
Keyword(s):  
International Space Station ◽  
Numerical Models ◽  
Space Station ◽  
Scale Model ◽  
Microgravity Environment ◽  
Small Scale ◽  
Piv Measurements ◽  
International Space ◽  
The Subject ◽  
Small Scale Model

An attempt at improving the ventilation solution for the crew quarters aboard the International Space Station requires a thorough understanding of the flow dynamics in a microgravity environment. An experimental study is required in order to validate the numerical models. As part of this process, a small-scale model was proposed for a detailed study of the velocity field. PIV measurements in water offer high quality results and were chosen for the subject. Following certain similitude criteria, an equivalence can be found between the results of these measurements and the real ventilation scenario. This paper describes the development process of this small-scale model as well as its performance in the initial test runs. Details regarding the advantages and weaknesses of this first model are the core of the paper, with the intention of aiding researchers in their design of similar models. The conclusion presents future steps and proposed improvements to the model.

Vibration Isolation in a Microgravity Environment

1993 ◽  
Vol 115 (4) ◽  
pp. 477-483
Author(s):  
R. M. Alexander ◽  
C. H. Gerhold ◽  
C. B. Atwood ◽  
J. F. Cordera
Keyword(s):  
Vibration Isolation ◽  
Center Of Mass ◽  
Space Station ◽  
Microgravity Environment ◽  
Isolation System ◽  
Air Jet ◽  
Surrounding Structure ◽  
Jet Control ◽  
Oscillatory Response ◽  
The Mathematical Model

Many in-space research experiments require the microgravity environment attainable near the center of mass of the proposed space station. Since dynamic disturbances to the surrounding structure may undermine an experiment’s validity, isolation of these experiments is imperative. This paper summarizes analytical and experimental work accomplished to develop an isolation system which allows the pay load to float freely within a prescribed boundary while being kept centered with forces generated by small jets of air. An experimental setup was designed and constructed to simulate the microgravity environment In the horizontal plane. Results demonstrate the air jet control system to be effective in managing payload oscillatory response. An analytical model was developed and verified by comparing predicted and measured payload response. The mathematical model is then used to investigate payload response to disturbances likely to be present in the space station.

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