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At least 19 records
All sourcesNASA NTRSDOE OSTI
NASA NTRS2014

Probiotics in the Space Food System: Delivery, Microgravity Effects, and the Potential Benefit to Crew Health

As mission distance and duration increase, the need grows for non‐invasive disease prevention and immunomodulation, especially given the limited medical response capability expected for these missions and the immune dysregulation documented in crew. Additionally, changes in diet, lifestyle, antibiotic usage, and the environmental stresses during spaceflight may alter crewmembers' intestinal microbiome. The addition of probiotic bacteria to the space food system is expected to confer immunostimulatory benefits on crewmembers, with the potential to counteract the immune dysregulation that has been documented in spaceflight. Based on previous studies that demonstrated unique microbiological responses to the low shear environment of spaceflight, probiotic organisms hold the potential to induce enhanced beneficial responses through mechanisms, such as beneficial interactions with human immune cells and repression of colonization of pathogens on the mucosa. The work presented here will begin to address two research gaps related to providing probiotics in spaceflight: 1) delivery, and 2) the effect of the low shear microgravity environment on probiotic attributes. The probiotic Lactobacillus acidophilus was selected for investigation due to its wide commercial use and documented benefits that include inhibition of virulence related gene expression in pathogens and mucosal stimulation of immune cells. The delivery system for probiotics has not been determined for spaceflight, where the food system is shelf stable and the lack of refrigeration prevents the use of traditional dairy delivery methods. In order to demonstrate the potential of the space food system to deliver viable probiotic bacteria to crewmembers, the probiotic L. acidophilus was packaged in high barrier flight packaging in nonfat dry milk (NFDM) or retained in commercial capsule form. Viable cells were enumerated over 8 months of storage at 22, 4, and ‐80ºC. The survival of L. acidophilus rehydrated in NFDM, in a PBS control, and directly from the capsule was also evaluated following stress challenge with simulated gastric and intestinal juices to determine the method that would deliver the most viable cells to the intestine, where they would be expected to confer beneficial effects. L. acidophilus was found to be stable to gastric and intestinal juice challenge when delivered in rehydrated NFDM, even after two hours of exposure. In comparison, L. acidophilus was reduced by 1‐5 logs when exposed to gastric and intestinal juice directly and when rehydrated in a PBS control. Shelf life data indicated that L. acidophilus would require refrigerated or frozen storage to remain viable at adequate levels over the multi‐year storage periods required for spaceflight. This study suggests that the protective effect provided by the dairy matrix, and not merely rehydration prior to consumption, extends probiotic viability and stress tolerance during storage in spaceflight and in simulated digestion conditions more adequately than a capsule. In addition to effective delivery, it is essential to understand the microgravity effects on the stress tolerances and genetic expression of probiotic bacteria to enable optimization of growth, survival, strain selection, and conferred benefits in spaceflight. In our current study, ground‐based characterization of the growth, stress response, and transcriptomic response of L. acidophilus will be accomplished using the low shear modeled microgravity (LSMMG) culture environment in the rotating wall vessel (RWV). We hypothesize that L. acidophilus will maintain or improve its growth and stress response following culture in the LSMMG environment, and that transcriptomic analysis will define the associated molecular mechanism(s), resulting in the ability to optimize strain selection.

Castro, S. L.↗
NASA NTRS2014

Delivery of Probiotics in the Space Food System

The addition of probiotic bacteria to the space food system is expected to confer immunostimulatory benefits on crewmembers during spaceflight, counteracting the immune dysregulation that has been documented in spaceflight [1]. Specifically, the probiotic Lactobacillus acidophilus has been shown to promote health benefits including antagonism towards and inhibition of virulence related gene expression in pathogens, mucosal stimulation of immune cells, and a reduction in the occurrence and duration of cold and flu-like symptoms [2-5]. The optimum delivery system for probiotics has not been determined for spaceflight, where the food system is shelf stable and the lack of refrigeration prevents the use of traditional dairy delivery methods. This work proposes to determine whether L. acidophilus is more viable, and therefore more likely to confer immune benefit, when delivered in a capsule form or when delivered in nonfat dry milk powder with a resuscitation opportunity upon rehydration, following 0, 4, and 8 months of storage at -80degC, 4degC, and 22degC, and both prior to and after challenge with simulated gastric and intestinal juices. We hypothesize that the low moisture neutral dairy matrix provided by the nonfat dry milk, and the rehydration step prior to consumption, will extend probiotic viability and stress tolerance compared to a capsule during potential storage conditions in spaceflight and in simulated digestion conditions.

Castro, S. L.↗
NASA NTRS2014

Delivery of Probiotics in the Space Food System

The addition of probiotic bacteria to the space food system is expected to confer immunostimulatory benefits on crewmembers during spaceflight, counteracting the immune dysregulation that has been documented in spaceflight. Specifically, the probiotic Lactobacillus acidophilus has been shown to promote health benefits including antagonism towards and inhibition of virulence related gene expression in pathogens, mucosal stimulation of immune cells, and a reduction in the occurrence and duration of cold and flu‐like symptoms. The optimum delivery system for probiotics has not been determined for spaceflight, where the food system is shelf stable and the lack of refrigeration prevents the use of traditional dairy delivery methods. This work proposes to determine whether L. acidophilus is more viable, and therefore more likely to confer immune benefit, when delivered in a capsule form or when delivered in nonfat dry milk powder with a resuscitation opportunity upon rehydration, following 0, 4, and 8 months of storage at ‐80degC, 4degC, and 22degC, and both prior to and after challenge with simulated gastric and intestinal juices. We hypothesize that the low moisture neutral dairy matrix provided by the nonfat dry milk, and the rehydration step prior to consumption, will extend probiotic viability and stress tolerance compared to a capsule during potential storage conditions in spaceflight and in simulated digestion conditions.

Castro, S. L.↗
NASA NTRS2016

Effect of Low Shear Modeled Microgravity (LSMMG) on the Probiotic Lactobacillus Acidophilus ATCC 4356

The introduction of generally recognized as safe (GRAS) probiotic microbes into the spaceflight food system has the potential for use as a safe, non-invasive, daily countermeasure to crew microbiome and immune dysregulation. However, the microgravity effects on the stress tolerances and genetic expression of probiotic bacteria must be determined to confirm translation of strain benefits and to identify potential for optimization of growth, survival, and strain selection for spaceflight. The work presented here demonstrates the translation of characteristics of a GRAS probiotic bacteria to a microgravity analog environment. Lactobacillus acidophilus ATCC 4356 was grown in the low shear modeled microgravity (LSMMG) orientation and the control orientation in the rotating wall vessel (RWV) to determine the effect of LSMMG on the growth, survival through stress challenge, and gene expression of the strain. No differences were observed between the LSMMG and control grown L. acidophilus, suggesting that the strain will behave similarly in spaceflight and may be expected to confer Earth-based benefits.

Stahl, S.↗
NASA NTRS2024

Augmentation of Seed Film Technology With Probiotics in Support of Space Crop Production

This project set out to establish a method for introducing Plant Growth Promoting Microbes to seed film for use in a flight-based crop production system. The inoculation method, demonstrated in this paper, succeeded in this introduction while simultaneously avoiding disrupting the integrity of the seed film material. Future research should continue to explore the idea of growth production and crop protection via intentional introduction of an exophytic microbiome including the verification of effects of varying inoculum concentrations on mature plants. Positive effects are more likely to be noted when multiple organisms are introduced simultaneously. Additional benefit is likely to be seen on an experimental basis when crops are challenged via the introduction of both biotic and abiotic stressors.

Aubrie E Orourke↗
NASA NTRS2015

2015 Advanced Environmental Health/Advanced Food Technology Standing Review Panel

The 2015 Advanced Environmental Health/Advanced Food Technology (AEH/AFT) Standing Review Panel (from here on referred to as the SRP) met for a site visit in Houston, TX on December 14 - 15, 2015. The SRP met with representatives from the Space Human Factors and Habitability (SHFH) Element and members of the Human Research Program (HRP) to review the updated research plans for the Risk of Adverse Health Effects Due to Host-Microorganism Interactions (MicroHost Risk) and the Risk of Performance Decrement and Crew Illness due to an Inadequate Food System (Food Risk). During the meeting, the SRP also met with the vehicle engineers to discuss possible food storage options. The SRP would like to commend Dr. Oubre and Dr. Douglas for their detailed presentations, as well the frank, refreshing, and comprehensive engineering presentation. This gave much needed perspective to the food storage issues and reassured the committee about NASA's approach to the problem. In terms of critiques, the SRP remains unconvinced about the rationale for probiotic use other than for specific applications supported by the literature. It is not clear what gap or problem is being addressed by the use of probiotics, and the rationale for their use needs to be clearly rooted in the available literature. The SRP thinks that if low-Earth orbit is associated with immune system impairment, then there may additional risks linked with the use of probiotics. It is not clear to the SRP how NASA will determine if probiotics are having their intended beneficial effect. A similar concern is raised as to what gaps or problems are being addressed by "functional foods". Mixed infections, rather than single species infections, which can augment severity of disease, also represent a significant concern. Overall, the SRP considers this to be a strong program that is well-organized, well-coordinated and generates valuable data.

Steinberg, Susan↗
NASA NTRSDate not supplied

The Future of Astronaut Nutrition: Daily Production of Kefir Nutrient Packs in a Lunar Analog Mission

Astronaut nutrition will need to be supplemented with fresh nutrients and probiotics for long duration space travel. Kefir is a fermented milk beverage that provides probiotic bacteria, protein, calcium, and vitamin K. It has been historically cultured for thousands of years and is a modern-day health food. During the BioNutrients-2 experiment, one generation of kefir was cultured by astronauts on the ISS in fluorinated ethylene propylene (FEP) bags. This follow-on experiment aimed to test the hypothesis that kefir can be produced at ambient temperatures and safely passaged to make a series of nutrient packs in an analog environment. The Hawaiian Space Exploration Analog and Simulation (HI-SEAS) is a semi-controlled habitat located on the Mauna Loa volcano on the island of Hawaii at 2,500 m elevation. The recent EMMIHS (EuroMoonMars, International MoonBase Alliance, HI-SEAS) missions comprised of interdisciplinary, international crews for mock lunar missions. During a six-day mission in March 2024, crew members tested continuous passaging of kefir cultures (C-FIR commercial strain). After approximately 24 hours of growth, a portion of culture was used to seed the next generation of culture bags for four passages. A color board and pH indicator allowed the crew to easily determine when the culture had reached optimal pH. Laboratory analysis at NASA Ames Research Center showed that the pH met Kefir standards (<5). The yeast and lactic acid bacteria were in normal range and no contamination was detected in the final passage. By successfully accomplishing this experiment the team has demonstrated the ability to safely produce daily cultures of kefir in a microgravity applicable growth system.

Astronaut nutrition↗
NASA NTRS2017

Development of Storage Methods for Saccharomyces Strains to be Utilized for In Situ Nutrient Production in Long-Duration Space Missions

Long-duration space missions will benefit from closed-loop life support technologies that minimize mass, volume, and power as well as decrease reliance on Earth-based resupply. A system for In situ production of essential vitamins and nutrients can address the documented problem of degradation of stored food and supplements. Research has shown that the edible yeast Saccharomyces cerevisiae can be used as an on-demand system for the production of various compounds that are beneficial to human health. A critical objective in the development of this approach for long-duration space missions is the effective storage of the selected microorganisms. This research investigates the effects of different storage methods on survival rates of the non-sporulating probiotic S. boulardii, and S. cerevisiae spores and vegetative cells. Dehydration has been shown to increase long-term yeast viability, which also allows increased shelf-life and reduction in mass and volume. The process of dehydration causes detrimental effects on vegetative cells, including oxidative damage and membrane disruption. To maximize cell viability, various dehydration methods are tested here, including lyophilization (freeze-drying), air drying, and dehydration by vacuum. As a potential solution to damage caused by lyophilization, the efficacy of various cryoprotectants was tested. Furthermore, in an attempt to maintain higher survival rates, the effect of temperature during long-term storage was investigated. Data show spores of the wild-type strain to be more resilient to dehydration-related stressors than vegetative cells of either strain, and maintain high viability rates even after one year at room temperature. In the event that engineering the organism to produce targeted nutrient compounds interferes with effective sporulation of S. cerevisiae, a more robust method for improving vegetative cell storage is being sought. Therefore, anhydrobiotic engineering of S. cerevisiae and S. boulardii is being conducted.

life support↗
NASA NTRS2013

Water Treatment Technologies Inspire Healthy Beverages

Mike Johnson, a former technician at Johnson Space Center, drew on his expertise as a wastewater engineer to create a line of kombucha-based probiotic drinks. Unpeeled Inc., based in Minneapolis-St. Paul, Minnesota, employs 12 people and has sold more than 6 million units of its NASA-inspired beverage.

Source record↗
NASA NTRS2017

Development of Storage Methods for Saccharomyces Strains to be Utilized for In situ Nutrient Production in Long-Duration Space Missions

Long-duration space missions will benefit from closed-loop life support technologies that minimize mass, volume, and power as well as decrease reliance on Earth-based resupply. A system for In situ production of essential vitamins and nutrients can address the documented problem of degradation of stored food and supplements. Research has shown that the edible yeast Saccharomyces cerevisiae can be used as an on-demand system for the production of various compounds that are beneficial to human health. A critical objective in the development of this approach for long-duration space missions is the effective storage of the selected microorganisms. This research investigates the effects of different storage methods on survival rates of the non-sporulating probiotic S. boulardii, and S. cerevisiae spores and vegetative cells. Dehydration has been shown to increase long-term yeast viability, which also allows increased shelf-life and reduction in mass and volume. The process of dehydration causes detrimental effects on vegetative cells, including oxidative damage and membrane disruption. To maximize cell viability, various dehydration methods are tested here, including lyophilization (freeze-drying), air drying, and dehydration by vacuum. As a potential solution to damage caused by lyophilization, the efficacy of various cryoprotectants was tested. Furthermore, in an attempt to maintain higher survival rates, the effect of temperature during long-term storage was investigated. Data show spores of the wild-type strain to be more resilient to dehydration-related stressors than vegetative cells of either strain, and maintain high viability rates even after one year at room temperature. In the event that engineering the organism to produce targeted nutrient compounds interferes with effective sporulation of S. cerevisiae, a more robust method for improving vegetative cell storage is being sought. Therefore, anhydrobiotic engineering of S. cerevisiae and S. boulardii is being conducted

nutrient↗
NASA NTRSDate not supplied

BioNutrients-1: Development of an On-Demand Nutrient Production System for Long-Duration Missions

Future long-duration missions beyond low-Earth orbit will require advances in food technologies to address the documented problem of degradation of vitamins and nutrients in supplied foods stored long-term. To begin to address the issue of nutrient degradation, we are developing and flight-testing a platform biomanufacturing technology for in situ production of target nutrients. This technology is being tested over a five-year duration on the International Space Station (ISS). As part of the BioNutrients-1 project we have developed an on-demand system for the production of two carotenoids, β-carotene and zeaxanthin, by genetically engineering distinct strains of Saccharomyces cerevisiae, more commonly known as baker’s yeast. The on-orbit nutrient production packs contain a desiccated yeast strain and edible growth substrate. Once hydrated, the contents of the production packs are intended to grow and produce a desired amount of ready-to-consume nutrients. In this current version the production packs will not be consumed and future missions will require an inactivation of microorganisms before consumption. In addition to the on-orbit hydration of the production packs a series of valuable microorganisms are currently being stored in stasis packs on the ISS including probiotics organisms, bacterial strains used in yogurt production, and organisms with potential use for future biomanufacturing. Analysis of returned ISS stasis packs and ground controls will include multi-omics studies and provide insight into long-term survival of organisms stored in a space environment. Both stasis packs and hydrated production packs will be intermittently returned to Earth for analysis. Preliminary data from long-term storage studies of stasis packs stored on the ISS for 47 days versus their ground control counterparts have shown no significant difference in viability. Currently no production packs have been processed.

Yeast↗
NASA NTRSDate not supplied

BioNutrients-2: Improvements to the BioNutrients-1 Nutrient Production System

The BioNutrients (BN) project aims to develop an on-demand nutrient production system capable of rapid and safe delivery of multiple nutrients in a single-use production pack for long-duration missions. The general concept of this system will be to develop an edible engineered microorganism capable of surviving during long-term storage, revival and growth of that microorganism for product production, and delivery of precise quantities of nutrients for human consumption. BN-1 is a five-year flight project currently on the International Space Station (ISS). It is testing the long-term storage of various microorganisms for the biomanufacturing of space relevant compounds and to validate the performance of the first generation (Gen-1) of production packs. The BN-2 project is a continuation of the BN-1 flight project and intends to expand the projects overall scope by introducing novel products, broadening the range of microbial food sources, and improving previously designed hardware from the BN-1 project. We are in the process of optimizing our platform biomanufacturing technology for in situ production of yogurt and kefir on the ISS. BN-2 will test the efficacy of this technology for future delivery of probiotic organisms which can be engineered to produce additional nutritional supplements (consumption by the crew is not planned in this project). Furthermore, to improve the feasibility of this technology for mission use we have transitioned from a hard-shell pack to a Gen-2, fluorinated ethylene propylene (FEP) bag. The single-use FEP bioreactor will facilitate growth of known organisms and ensure a contaminant free environment, while reducing mass and volume. These bioreactors will be employed for production of additional space relevant products in the BN-2 project. On-orbit testing of Gen-2 production packs will be necessary for future iterations, in order to optimize bag design and product production for eventual flight use by the crew.

BioNutrients↗
NASA NTRSDate not supplied

BioNutrients-2: Improvements to the BioNutrients-1 Nutrient Production System

The BioNutrients (BN) project aims to develop an on-demand biological production system capable of rapid and safe delivery of multiple nutrients in single-use production packs for long-duration missions. The general concept of this system is to develop edible engineered microorganisms that produce nutrients for human consumption following long-term storage, revival, and growth. BN-1 is a five-year flight project currently on the International Space Station (ISS). It is testing the long-term storage of various microorganisms for the biomanufacturing of space-relevant compounds and validating the performance of the first generation of production packs. The BN-2 project expands the BN-1 flight project scope by introducing novel products, broadening the range of microbial food sources, and improving upon production pack hardware. We are optimizing the biomanufacturing platform for in situ production of yogurt and kefir on the ISS. BN-2 also will test engineered microorganisms, including a probiotic strain, to produce carotenoids or follistatin. To improve the feasibility of this technology for mission use, we transitioned from a hard-shell pack to a second-generation fluorinated ethylene propylene (FEP) bag. The single-use FEP bioreactor facilitates growth of target organisms in a contaminant free environment, while reducing mass and volume compared to the BN-1 hardware. These bioreactors will be employed for production of additional space-relevant products in the BN-2 project. On-orbit testing of FEP production packs will enable optimization of the platform for eventual flight use by the crew.

Natalie Ball↗
NASA NTRSDate not supplied

BioNutrients: Microbial on-demand production of short shelf-life micronutrients in space

Long duration deep space missions pose a significant challenge when considering adequate nutrition for human spaceflight. Many micronutrients supplied in pre-packaged foods for low Earth orbit flights have a short shelf life and will degrade over multi-year missions. One strategy to mitigate the risk of micronutrient malnutrition is to grow fresh foods to supplement a pre-packaged diet. The BioNutrients project aims to develop systems for growing edible microorganisms that are genetically engineered to produce short shelf life micronutrients. BioNutrients-1 (BN-1) is a five-year spaceflight project currently underway on the International Space Station (ISS) that tests long-term storage and production of two carotenoids, β-carotene and zeaxanthin, in genetically engineered yeast strains. BN-1 on-orbit and ground control activities indicate that the nutrient production system will be stable for at least two years in ambient storage conditions. BN-1 is also testing the effects of long-term ISS storage on viability and gene expression using “stasis packs” containing a series of microorganisms useful in food and biomanufacturing processes. Stasis pack analysis is distinguishing microorganisms with full viability after two years of storage from other species that lose viability rapidly. In parallel, BioNutrients-2 (BN-2) is being developed as a six-month flight experiment that will expand the range of foods produced on ISS to include yogurt, kefir, and probiotic yeast. BN-2 will improve upon hardware design by transitioning from a hard-shell polycarbonate bioreactor to a flexible fluorinated ethylene propylene (FEP) bag. This change reduces the mass and volume required to store production packs. The long-term goal of BioNutrients is to develop the capability for rapid and safe delivery of multiple micronutrients in single-use production packs. We expect synthetic biology approaches similar to BioNutrients will play an important role for astronaut health in future NASA missions to the Moon and Mars.

Synthetic Biology↗
NASA NTRSDate not supplied

Assessment of Individualizing lactobacillus plantarum Supplementation with Precision Health to Preserve Muscle Health in Astronauts During Long Duration Spaceflight

Muscle atrophy is an unfortunate reality seen after spending time in microgravity. Decreases in muscle strength, size, and endurance can negatively impact mission productivity and increase the risk of injury in astronauts. In preparing for long-duration spaceflight aboard future orbiting stations and interplanetary vessels, supplementation with lactobacillus plantarum has been suggested as a possible countermeasure for crew members. Early clinical trials have shown the probiotic to improve muscle strength, endurance, and associated biomarkers, even without significant physical exercise by influencing the gut-muscle-axis. While current onboard exercise protocols have significantly helped limit degradation (as seen below), individualized implementation of this supplement could not only add another layer of protection but also protect against inadequate exercise equipment on future missions due to payload constraints

Precision Health↗
NASA NTRSDate not supplied

Validation of Multisystem Countermeasures Protocol for Spaceflight during Antarctica Winter-over at Palmer Station (Palmer Countermeasures)

Stressors associated with spaceflight induce persistent immune compromise in astronauts which increase subclinical latent virus reactivation. In select crews, adverse clinical events have been documented. Antarctica winter-over (AWO) mission most closely reproduces these mission stressors: prolonged deployment, extreme environment, circadian misalignment, isolation, station lifestyle, and personal risk. The US maintains three primary stations in Antarctica: South Pole Station, McMurdo, and Palmer. Previous studies suggest that stations located near the interior of Antarctica (South Pole, McMurdo) have confounding effects on the immune system due to persistent hypobaric hypoxia. We hypothesized that winter-over at a coastal station (Palmer) would be more akin to spaceflight due to its normoxic but still extreme environment. Therefore, AWO at Palmer Station was selected, and validated in a pilot study [2], as the platform for testing and validating the effectiveness of an immune-restorative countermeasure protocol designed for deep space missions. Specifics include diet modifications, nutritional supplementation (vitamin D, probiotic, etc.), prescribed aerobic and resistive exercise, and a protocol of stress relieving virtual reality exercises. A multitude of biological sample types, including blood, saliva, and hair will be collected in tandem with the countermeasures in order to examine the combined effectiveness of the countermeasures. Samples and logs from subjects will be transported from Palmer Station to Johnson Space Center for further processing and distribution to co-investigators at the end of each winter-over. Extracted samples will be analyzed by appropriate testing platforms (Multiplex, qPCR, ELISA, etc.) to monitor alterations in leukocyte distribution, T cell and NK function, cytokine profiles, reactivation of latent herpesviruses, and nutritional factors. The data collected will be compared to a control year in which no countermeasures were deployed to evaluate the overall effectiveness of the analog and to validate the candidate immune countermeasure strategy. AWO 2023 concluded with the 4th in-mission timepoint conducted in September 2023. Samples for 16 subjects, including blood, saliva, hair, surveys, and PCR data, were all successfully returned from Antarctica to NASA/JSC mid-November 2023. Samples have since been distributed to co-investigators for further processing and analysis. With the completion of the first countermeasure year, preliminary data on the effectiveness of the deep-space protocol is being evaluated, however, no conclusions can be drawn yet until the completion of the second AWO countermeasure year, AWO 2024. AWO 2024 commenced in late-March 2024, with 13 subjects consenting and performing their baseline data collections (BDCs). Unique to the 2024 deployment, NSF lifted certain COVID restrictions and rallied the crewmembers in Punta Arenas, Chile. All NSF activities were transferred to this location and NASA was allowed, for the first time, to perform consent briefings, baseline samplings and training in person. This augment greatly increased the likelihood of success for the overwinter activities.

Cody L Gutierrez↗