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At least 19 records

What is microbial dormancy?

Life can be stressful. One way to deal with stress is to simply wait it out. Microbes do this by entering a state of reduced activity and increased resistance commonly called ‘dormancy’. But what is dormancy? Different scientific disciplines emphasize distinct traits and phenotypic ranges in defining dormancy for their microbial species and system-specific questions of interest. Here, we propose a unified definition of microbial dormancy, using a broad framework to place earlier discipline-specific definitions in a new context. We then discuss how this new definition and framework may improve our ability to investigate dormancy using multi-omics tools. Finally, we leverage our framework to discuss the diversity of genomic mechanisms for dormancy in an extreme environment that challenges easy definitions – the permafrost.

59 BASIC BIOLOGICAL SCIENCES↗

Ground Test Validations for Uncrewed Dormancy Preparation and Recovery

In future long duration space flight and habitation missions, periods of uncrewed dormancy where life support systems are shutdown or in low power-modes will occur. To examine the impacts of these dormant periods, along with potential pre- and post-dormancy procedures, Marshall Space Flight Center has been building an uncrewed dormancy test stand. The uncrewed dormancy test stand will include a urine separator from the Universal Waste Management System, a condensate separator, urine filter, brine filter and distillate filter. In conjunction to the test stand, the Center for Biofilm Engineering (CBE) has ongoing long-term dormancy experiments to assess microbial survival in a variety of simulated shutdown procedures including shut-in, flushing with and without biocide, and dry-out conditions. This publication will serve as a discussion of dormancy considerations, criteria for success, and experimental designs for testing dormancy conditions and shutdown procedures. It will include an update of ongoing test stand construction, including a description of the current lower-fidelity versions of the urine and condensate separators being produced. It will also examine dormancy work being conducted by the CBE and discuss how this may factor into procedures tested on the uncrewed dormancy test stand.

Eric Beitle↗

Microbial Adaptation to Spaceflight Is Correlated With Bacteriophage-Encoded Functions

Evidence from the International Space Station suggests microbial populations are rapidly adapting to the spacecraft environment; however, the mechanism of this adaptation is not understood. Bacteriophages are prolific mediators of bacterial adaptation on Earth. Here we survey 245 genomes sequenced from bacterial strains isolated on the International Space Station for dormant (lysogenic) bacteriophages. Our analysis indicates phage-associated genes are significantly different between spaceflight strains and their terrestrial counterparts. Additionally, we identify 283 complete prophages, those that could initiate bacterial lysis and infect additional hosts, of which 21% are novel. These prophage regions encode functions that correlate with increased persistence in extreme environments, such as spaceflight, to include antimicrobial resistance and virulence, DNA damage repair, and dormancy. Our results correlate microbial adaptation in spaceflight to bacteriophage-encoded functions that may impact human health in spaceflight.

Space Biology↗

AMEBoP: Additive Manufacturing Enabled Biofilm Prevention

Objective: Leverage additive manufacturing to fortify Environmental Control and Life Support Systems (ECLSS) manifolds and other vulnerable components against biofilm growth during dormancy by eliminating dead legs to reduce total stagnate fluid volume and available nutrients and by printing from inherently biocidal materials.

ECLSS↗

AMEBoP: Additive Manufacturing Enabled Biofilm Prevention

Leverage additive manufacturing to fortify ECLSS (Environmental Control and Life Support Systems) manifolds and other vulnerable components against biofilm growth during dormancy. This project aims to improve dormancy tolerance by eliminating dead legs to reduce stagnate fluid volume and to limit available nutrients, and by printing from inherently biocidal materials.

ECLSS↗

Water Recovery System Design to Accommodate Dormant Periods for Manned Missions

Future manned missions beyond lower Earth orbit may include intermittent periods of extended dormancy. Under the NASA Advanced Exploration System (AES) project, NASA personnel evaluated the viability of the ISS Water Recovery System (WRS) to support such a mission. The mission requirement includes the capability for life support systems to support crew activity, followed by a dormant period of up to one year, and subsequently for the life support systems to come back online for additional crewed missions. Dormancy could be a critical issue due to concerns with microbial growth or chemical degradation that might prevent water systems from operating properly when the crewed mission began. As such, it is critical that the water systems be designed to accommodate this dormant period. This paper details the results of this evaluation, which include identification of dormancy issues, results of testing performed to assess microbial stability of pretreated urine during dormancy periods, and concepts for updating to the WRS architecture and operational concepts that will enable the ISS WRS to support the dormancy requirement.

Tabb, David↗

Silver Electrolysis for Disinfection of Spacecraft Potable Water: 2024 Update

Anodic dissolution of silver electrodes, or “silver electrolysis,” is being investigated as a means of introducing biocidal silver into potable water on exploration spacecraft. This paper provides an update on the effort to implement this technology into a spacecraft potable water system. Previous papers reported on the feasibility of the technology for this application, strategies to prevent a potential fault condition termed “electrode bridging,” results from a preliminary investigation into the cathode reaction, and preliminary multiphysics modeling of the reactor. Since then, work has begun on the design of a next-generation silver electrolysis reactor prototype that will incorporate improvements identified in previous testing and package the reactor in a more flight-like configuration. This development effort has included additional testing to optimize the reactor design for prevention of electrode bridging, further investigation into the cathode reaction (including the use of a dissolved hydrogen sensor), and an assessment of the feasibility of using the reactor in other applications, such as for microbial shock and preparation for system dormancy.

silver biocide↗

Silver Electrolysis for Disinfection of Spacecraft Potable Water: 2024 Update

Anodic dissolution of silver electrodes, or “silver electrolysis,” is being investigated as a means of introducing biocidal silver into potable water on exploration spacecraft. This paper provides an update on the effort to implement this technology into a spacecraft potable water system. Previous papers reported on the feasibility of the technology for this application, strategies to prevent a potential fault condition termed “electrode bridging,” results from a preliminary investigation into the cathode reaction, and preliminary multiphysics modeling of the reactor. Since then, work has begun on the design of a next-generation silver electrolysis reactor prototype that will incorporate improvements identified in previous testing and package the reactor in a more flight-like configuration. This development effort has included additional testing to optimize the reactor design for prevention of electrode bridging, further investigation into the cathode reaction (including the use of a dissolved hydrogen sensor), and an assessment of the feasibility of using the reactor in other applications, such as for microbial shock and preparation for system dormancy.

silver biocide↗

Water Recovery System Architecture and Operational Concepts to Accommodate Dormancy

Future manned missions beyond low Earth orbit will include intermittent periods of extended dormancy. The mission requirement includes the capability for life support systems to support crew activity, followed by a dormant period of up to one year, and subsequently for the life support systems to come back online for additional crewed missions. NASA personnel are evaluating the architecture and operational concepts that will allow the Water Recovery System (WRS) to support such a mission. Dormancy could be a critical issue due to concerns with microbial growth or chemical degradation that might prevent water systems from operating properly when the crewed mission began. As such, it is critical that the water systems be designed to accommodate this dormant period. This paper identifies dormancy issues, concepts for updating the WRS architecture and operational concepts that will enable the WRS to support the dormancy requirement.

Carter, Layne↗

Dormancy and Recovery Testing for Biological Wastewater Processors

Bioreactors, such as the aerated hollow fiber membrane type, have been proposed and studied for a number of years as an alternate approach for treating wastewater streams for space exploration. Several challenges remain to be resolved before these types of bioreactors can be used in space settings, including transporting the bioreactors with intact and active biofilms, whether that be to the International Space Station or beyond, or procedures for safing the systems and placing them into a dormant state for later start-up. Little information is available on such operations as it is not common practice for terrestrial systems. This study explored several dormancy processes for established bioreactors to determine optimal storage and recovery conditions. Procedures focused on complete isolation of the microbial communities from an operational standpoint and observing the effects of: 1) storage temperature, and 2) storage with or without the reactor bulk fluid. The first consideration was tested from a microbial integrity and power consumption standpoint; both ambient temperature (25 C) and cold (4 C) storage conditions were studied. The second consideration was explored; again, for microbial integrity as well as plausible real-world scenarios of how terrestrially established bioreactors would be transported to microgravity and stored for periods of time between operations. Biofilms were stored without the reactor bulk fluid to simulate transport of established biofilms into microgravity, while biofilms stored with the reactor bulk fluid simulated the most simplistic storage condition to implement operations for extended periods of nonuse. Dormancy condition did not have an influence on recovery in initial studies with immature biofilms (48 days old), however a lengthy recovery time was required (20+ days). Bioreactors with fully established biofilms (13 months) were able to recover from a 7-month dormancy period to steady state operation within 4 days (approx. 1 residence cycle). Results indicate a need for future testing on biofilm age and health and further exploration of dormancy length.

Recovery Testing↗

Climate history modulates stress responses of common soil bacteria under experimental drought

Soil drying challenges microbial viability and survival, with bacteria employing various mechanisms to respond to shifts in osmolarity, including dormancy or metabolic upregulation of osmoprotectants. However, the extent to which these responses are shaped by an organism's phylogeny, or the climate history of a given environment is poorly understood. This study examines the responses of phylogenetically similar bacteria from semi-arid and humid tropical forest soils to osmotic and matric stress using synchrotron radiation-based Fourier Transform Infrared spectromicroscopy. This non-destructive approach depicts the biochemical phenotype for whole cells under control and stress conditions. We observed that, under osmotic stress, bacteria upregulated cell-signaling pathways, rapidly turned over lipid-storage compounds, and increased osmolyte production. In contrast, matric stress induced a more muted response, typically elevating the production of carbohydrate stress compounds, such as glycine betaine and trehalose. Whereas phylogenetically similar bacteria showed comparable biochemistry under control conditions, climate history played an important role in regulating responses to stress, whereby a stronger metabolic response was observed from semi-arid relative to tropical forest isolates. We conclude that bacterial stress response to drought can be more diverse than previously observed and regulated by both phylogeny and climate history.

54 ENVIRONMENTAL SCIENCES↗

Dormancy and Recovery Testing for Biological Wastewater Processors

Bioreactors, such as aerated membrane type bioreactors have been proposed and studied for a number of years as an alternate approach for treating wastewater streams for space exploration. Several challenges remain before these types of bioreactors can be used in space settings, including transporting the bioreactors with their microbial communities to space, whether that be the International Space Station or beyond, or procedures for safing the systems and placing them into dormant state for later start-up. Little information is available on such operations as it is not common practice for terrestrial systems. This study explored several dormancy processes for established bioreactors to determine optimal storage and recovery conditions. Procedures focused on complete isolation of the microbial communities from an operational standpoint and observing the effects of: 1) storage temperature, and 2) storage with or without the reactor bulk fluid. The first consideration was tested from a microbial integrity and power consumption standpoint; both room temperature (25 C) and cold (4 C) storage conditions were studied. The second consideration was explored; again, for microbial integrity as well as plausible real-world scenarios of how terrestrially established bioreactors would be transported to microgravity and stored for periods of time between operations. Biofilms were stored without the reactor bulk fluid to simulate transport of established biofilms into microgravity, while biofilms stored with the reactor bulk fluid simulated the most simplistic storage condition to implement operations for extended periods of nonuse. Dormancy condition did not have an influence on recovery in initial studies with immature biofilms (48 days old), however, a lengthy recovery time was required (20+ days). Bioreactors with fully established biofilms (13 months) were able to recover from a 7-month dormancy period to steady state operation within 4 days (approximately 1 residence cycle). Results indicate a need for future testing on biofilm age and health and further exploration of dormancy length.

Dormancy↗

Impact of Biofilms on the Design and Operation of ISS Life Support Systems

Biofilm growth has been an ongoing issue for US and Russian water systems on the International Space Station, and is a critical issue for exploration missions in which water systems must be designed to accommodate dormant periods of up to one year. On ISS, Russian condensate plumbing has previously clogged with biomass, requiring condensate plumbing to now be regularly replaced. In the US Segment, the release of biofilm from the Water Processor waste tank has clogged a solenoid valve downstream of the tank, resulting in the costly replacement of the inlet separator and process pump. Subsequent management of the biofilm in the waste tank involves restrictions on tank cycles to limit the release of biomass and an additional filter to protect downstream components. Engineering personnel are now evaluating concepts to better manage the biomass, including the use of microbial inhibitors and UV LEDs. Though current ISS operations could likely be sustained for the duration of ISS, a more effective method must be developed for managing the growth and release of biomass in future exploration vehicles. Biofilm management for future missions is complicated by the requirement to accommodate extended periods of dormancy during which time the water system will be stagnant. The current approach under consideration is to flush the waste water with product water to reduce the organic content followed by use of microbial inhibitors or UV. However, other concepts may also be developed based on ongoing research.

Carter, Donald Layne↗

Time-series RNA metabarcoding of the active Populus tremuloides root microbiome reveals hidden temporal dynamics and dormant core members

The rhizosphere is a critical interface between plant roots and soil, harboring diverse microbial communities that are essential to plant and ecosystem health. Although these communities exhibit stark temporal dynamics, their dormancy/activity transitions remain poorly understood. Such transitions may enable microbes to rapidly adjust functional contributions faster than community turnover alone would allow. Here, we used RNA metabarcoding to characterize the active fraction of microbial communities on the roots of quaking aspen (Populus tremuloides) in a time-series study across a natural environmental gradient. We explore cryptic temporal microbial community dynamics of rhizosphere communities at the ecosystem scale. The active rhizosphere bacterial and fungal communities were more temporally dynamic than total communities, while total communities exhibited a stronger response to site-specific conditions. Notably, some core microbiome members were often inactive, yielding a smaller “active core” subset. The fungal endophyte Hyaloscypha finlandica was the only microbe that was both present and active in all plots across all timepoints. Soil temperature strongly influenced both total and active community composition, with the fungal class Eurotiomycetes showing a temperature-dependent seasonal decline in abundance. Together, these results reveal that modulation of microbial activity levels is a key mechanism by which the plant root holobiont responds to environmental variation, and that even dominant symbionts may frequently persist in dormancy within the rhizosphere.

Community Structure and Diversity↗

To Biocide or not to Biocide? Exploring the "No Biocide" Option in Spacecraft Potable Water Systems

Residual biocide has been used to protect against microbial growth in spacecraft potable water systems since the Gemini program. Iodine, the biocide currently used on the International Space Station (ISS) has a long history of use but presents a few challenges for exploration: it must be removed using a consumable absorbent before crew consumption and is known to lose its biocidal concentration due to interactions with wetted components, especially during periods of dormancy. Biocidal silver is being considered for exploration missions as it is safe for crew consumption, but it too has challenges with proper material design to maintain sufficient disinfection residuals. An idea proposed by some in the Environmental Control and Life Support System (ECLSS) community has been that the biocide does not provide a true barrier against microbial growth, and therefore, perhaps the challenges of maintaining a residual biocide in the system could be best overcome by removing the biocide altogether. In this argument, they point to the regenerative water treatment system on the ISS which produces high purity water that theoretically meets the microbiological requirements before the iodine is added. Exploration water processing systems are expected to have similar water processing capabilities. This paper will discuss the role that residual biocide may play in controlling microbial growth and presents perspectives on the potential risks associated with removing it from exploration water systems.

biocide↗

To Biocide or not to Biocide? Exploring the "No Biocide" Option in Spacecraft Potable Water Systems

Residual biocide has been used to protect against microbial growth in spacecraft potable water systems since the Gemini program. Iodine, the biocide currently used on the International Space Station (ISS) has a long history of use but presents a few challenges for exploration: it must be removed using a consumable absorbent before crew consumption and is known to lose its biocidal concentration due to interactions with wetted components, especially during periods of dormancy. Biocidal silver is being considered for exploration missions as it is safe for crew consumption, but it too has challenges with proper material design to maintain sufficient disinfection residuals. An idea proposed by some in the Environmental Control and Life Support System (ECLSS) community has been that the biocide does not provide a true barrier against microbial growth, and therefore, perhaps the challenges of maintaining a residual biocide in the system could be best overcome by removing the biocide altogether. In this argument, they point to the regenerative water treatment system on the ISS which produces high purity water that theoretically meets the microbiological requirements before the iodine is added. Exploration water processing systems are expected to have similar water processing capabilities. This paper will discuss the role that residual biocide may play in controlling microbial growth and presents perspectives on the potential risks associated with removing it from exploration water systems.

biocide↗

To Biocide or Not to Biocide? Exploring the "No Biocide" Option in Spacecraft Potable Water Systems

Residual biocide has been used to protect against microbial growth in spacecraft potable water systems since the Gemini program. Iodine, the biocide currently used on the International Space Station (ISS) has a long history of use but presents a few challenges for exploration: it must be removed using a consumable absorbent before crew consumption and is known to lose its biocidal concentration due to interactions with wetted components, especially during periods of dormancy. Biocidal silver is being considered for exploration missions as it is safe for crew consumption, but it too has challenges with proper material design to maintain sufficient disinfection residuals. An idea proposed by some in the Environmental Control and Life Support System (ECLSS) community has been that the biocide does not provide a true barrier against microbial growth, and therefore, perhaps the challenges of maintaining a residual biocide in the system could be best overcome by removing the biocide altogether. In this argument, they point to the regenerative water treatment system on the ISS which produces high purity water that theoretically meets the microbiological requirements before the iodine is added. Exploration water processing systems are expected to have similar water processing capabilities. This paper will discuss the role that residual biocide may play in controlling microbial growth and presents perspectives on the potential risks associated with removing it from exploration water systems.

biocide↗

To Biocide or Not to Biocide? Exploring the "No Biocide" Option in Spacecraft Potable Water Systems

Residual biocide has been used to protect against microbial growth in spacecraft potable water systems since the Gemini program. Iodine, the biocide currently used on the International Space Station (ISS) has a long history of use but presents a few challenges for exploration: it must be removed using a consumable absorbent before crew consumption and is known to lose its biocidal concentration due to interactions with wetted components, especially during periods of dormancy. Biocidal silver is being considered for exploration missions as it is safe for crew consumption, but it too has challenges with proper material design to maintain sufficient disinfection residuals. An idea proposed by some in the Environmental Control and Life Support System (ECLSS) community has been that the biocide does not provide a true barrier against microbial growth, and therefore, perhaps the challenges of maintaining a residual biocide in the system could be best overcome by removing the biocide altogether. In this argument, they point to the regenerative water treatment system on the ISS which produces high purity water that theoretically meets the microbiological requirements before the iodine is added. Exploration water processing systems are expected to have similar water processing capabilities. This paper will discuss the role that residual biocide may play in controlling microbial growth and presents perspectives on the potential risks associated with removing it from exploration water systems.

biocide↗