Regenerative Life Support Systems Research Articles

Dependability model of automated intelligent regenerative life support system for space missions

Long-duration human space missions require intelligent regenerative life support systems that can recycle resources and automatically manage failures. This paper explores using Petri nets to model the reliability and complex interactions of such closed-loop systems. An architecture consisting of primary systems, backups, and consumable reserves is outlined. The automation system that controls everything is described. Petri nets can capture concurrency, failure modes, redundancy, and dynamic behavior. A modular modeling methodology is presented to develop hierarchical Petri net models that scale in fidelity. Elementary fragments represent failures and redundancy. Subsystem modules can be substituted for more detailed models. Analysis and simulation assess system reliability and failure response. This supports designing ultra-reliable systems to safely sustain human life in space.

Whole transcriptome analysis highlights nutrient limitation of nitrogen cycle bacteria in simulated microgravity

Regenerative life support systems (RLSS) will play a vital role in achieving self-sufficiency during long-distance space travel. Urine conversion into a liquid nitrate-based fertilizer is a key process in most RLSS. This study describes the effects of simulated microgravity (SMG) on Comamonas testosteroni, Nitrosomonas europaea, Nitrobacter winogradskyi and a tripartite culture of the three, in the context of nitrogen recovery for the Micro-Ecological Life Support System Alternative (MELiSSA). Rotary cell culture systems (RCCS) and random positioning machines (RPM) were used as SMG analogues. The transcriptional responses of the cultures were elucidated. For CO2-producing C. testosteroni and the tripartite culture, a PermaLifeTM PL-70 cell culture bag mounted on an in-house 3D-printed holder was applied to eliminate air bubble formation during SMG cultivation. Gene expression changes indicated that the fluid dynamics in SMG caused nutrient and O2 limitation. Genes involved in urea hydrolysis and nitrification were minimally affected, while denitrification-related gene expression was increased. The findings highlight potential challenges for nitrogen recovery in space.

Continuous cultivation of microalgae yields high nutrient recovery from nitrified urine with limited supplementation

Microalgae can play a key role in the bioeconomy, particularly in combination with the valorisation of waste streams as cultivation media. Urine is an example of a widely available nutrient-rich waste stream, and alkaline stabilization and subsequent full nitrification in a bioreactor yields a stable nitrate-rich solution. In this study, such nitrified urine served as a culture medium for the edible microalga Limnospira indica. In batch cultivation, nitrified urine without additional supplements yielded a lower biomass concentration, nutrient uptake and protein content compared to modified Zarrouk medium, as standard medium. To enhance the nitrogen uptake efficiency and biomass production, nitrified urine was supplemented with potentially limiting elements. Limited amounts of phosphorus (36 mg L−1), magnesium (7.9 mg L−1), calcium (12.2 mg L−1), iron (2.0 mg L−1) and EDTA (88.5 mg Na2-EDTA.2H2O L−1) rendered the nitrified urine matrix as effective as modified Zarrouk medium in terms of biomass production (OD750 of 1.2), nutrient uptake (130 mg N L−1) and protein yield (47%) in batch culture. Urine precipitates formed by alkalinisation could in principle supply enough phosphorus, calcium and magnesium, requiring only external addition of iron, EDTA and inorganic carbon. Subsequently, the suitability of supplemented nitrified urine as a culture medium was confirmed in continuous Limnospira cultivation in a CSTR photobioreactor. This qualifies nitrified urine as a valuable and sustainable microalgae growth medium, thereby creating novel nutrient loops on Earth and in Space, i.e., in regenerative life support systems for human deep-space missions.

Biofilm reactors for the treatment of used water in space:potential, challenges, and future perspectives

Water is not only essential to sustain life on Earth, but also is a crucial resource for long-duration deep space exploration and habitation. Current systems in space rely on the resupply of water from Earth, however, as missions get longer and move farther away from Earth, resupply will no longer be a sustainable option. Thus, the development of regenerative reclamation water systems through which useable water can be recovered from “waste streams” (i.e., used waters) is sorely needed to further close the loop in space life support systems. This review presents the origin and characteristics of different used waters generated in space and discusses the intrinsic challenges of developing suitable technologies to treat such streams given the unique constrains of space exploration and habitation (e.g., different gravity conditions, size and weight limitations, compatibility with other systems, etc.). In this review, we discuss the potential use of biological systems, particularly biofilms, as possible alternatives or additions to current technologies for water reclamation and waste treatment in space. The fundamentals of biofilm reactors, their advantages and disadvantages, as well as different reactor configurations and their potential for use and challenges to be incorporated in self-sustaining and regenerative life support systems in long-duration space missions are also discussed. Furthermore, we discuss the possibility to recover value-added products (e.g., biomass, nutrients, water) from used waters and the opportunity to recycle and reuse such products as resources in other life support subsystems (e.g., habitation, waste, air, etc.).

Design and implementation of regenerative life support system in the China space station

<p indent="0mm">China’s Tiangong space station is equipped with a regenerative life support system (RLSS). RLSS enables in-orbit environmental control and life support by producing oxygen, purifying air, recovering water from urine and condensate water. As a key technology of the space station, the RLSS contains several physicochemical systems, including electrolysis oxygen generation, carbon dioxide removal, trace contaminant control, urine processing, water recovery, and carbon dioxide reduction systems. During the Shenzhou-12 mission, the RLSS performed a 90-day manned operation and conducted the closed-loop recycling of the oxygen and water. This article presents the details pertaining to the techniques, developments, and operations of the RLSS on the China space station.

Metaproteomics, Heterotrophic Growth, and Distribution of Nitrosomonas europaea and Nitrobacter winogradskyi after Long-Term Operation of an Autotrophic Nitrifying Biofilm Reactor

Bioregenerative life support systems (BLSS) are currently in development to tackle low recovery efficiencies, high energy demands, as well as food, water, and oxygen production challenges through the regeneration of nutrients from waste streams. The MELiSSA pilot plant has been developed as a testbed for regenerative life support system bioreactor operation and characterization. As nitrogen is a vital resource in such systems, we studied the functional composition of a new packed-bed nitrifying bioreactor inoculated with a co-culture of Nitrosomonas europaea (ATCC 25978) and Nitrobacter winogradskyi (ATCC 25391). After 840 days of autotrophic continuous cultivation, the packed-bed was sampled at five vertical positions, each with three horizontal positions, and the biomass at each position was characterized via qPCR, 16S amplicon sequencing, and liquid chromatography tandem mass spectrometry. The total number of cells within the different sections fluctuated around 8.95 ± 5.10 × 107 cells/mL of beads. Based on 16S amplicons and protein content, N. europaea and N. winogradskyi constituted overall 44.07 ± 11.75% and 57.53 ± 12.04% of the nitrifying bioreactor, respectively, indicating the presence of a heterotrophic population that, even after such a long operation time, did not affect the nitrification function of the bioreactor. In addition, DNA-based abundance estimates showed that N. europaea was slightly more abundant than N. winogradskyi, whereas protein-based abundance estimates indicated a much higher abundance of N. europaea. This highlights that single-method approaches need to be carefully interpreted in terms of overall cell abundance and metabolic activity.

USE OF METHANE IN CLOSED-LOOP LIFE SUPPORT SYSTEMS FOR SPACE MISSIONS

The paper discusses the use of methane (generated in the process of oxygen recovery from carbon dioxide released by the crew during its hydrogenation in the Sabatier reaction, with subsequent extraction of 61% of oxygen through electrolysis of the resultant water) in a regenerative life support system for crews on space missions. It demonstrates that the methane resulting from Sabatier reaction can be used both for pyrolysis in order to return the resulting hydrogen into this reaction so as to extract 100% of oxygen from carbon dioxide, and for producing food protein for life support in space. The use of methane pyrolysis was enabled by new technologies which allowed lowering the process temperature down to 500–700°C and obtaining the easy-to-remove carbon. It provides recommendations for designing space systems for methane pyrolysis. The paper makes the case for use of the existing processes for industrial production of protein from methane using methanotrophic bacteria in the production of food protein for space food rations, determines the balance of a closed-loop methanotrophic reaction, provides calculation basis and recommendations for designing space systems for methanotrophic production of food protein. Development of a system for food protein production from methane will enable its use as one of the systems for providing food on the Moon and Mars, as well as a backup system in space transportation missions. Key words: space missions, crew life support, СО2 hydration, methane pyrolysis, methanotrophic bacteria, food protein.

Elliptic Array Luminescent Solar Concentrators for Combined Power Generation and Microalgae Growth

The full utilization of broadband solar irradiance is becoming increasingly useful for applications such as long-term space missions, wherein power generation from external sources and regenerative life support systems are essential. Luminescent solar concentrators (LSCs) can be designed to separate sunlight into photosynthetically active radiation (PAR) and non-PAR to simultaneously provide for algae cultivation and electric power generation. However, the efficiency of LSCs suffers from high emission losses. In this work, we show that by shaping the LSC in the form of an elliptic array, rather than the conventional planar configuration, emission losses can be drastically reduced to the point that they are almost eliminated. Numerical results, considering the combined effects of emission, transmission and surface scattering losses show the optical efficiency of the elliptic array LSC is 63%, whereas, in comparison, the optical efficiency for conventional planar LSCs is 47.2%. Further, results from numerical simulations show that elliptic array luminescent solar concentrators can convert non-PAR and green-PAR to electric power with a conversion efficiency of ~17% for AM1.5 and 17.6% for AM0, while transmitting PAR to an underlying photobioreactor to support algae cultivation.

Ground Demonstration of the Use of Limnospira indica for Air Revitalization in a Bioregenerative Life-Support System Setup: Effect of Non-Nitrified Urine–Derived Nitrogen Sources

Long-duration human space missions require considerable amounts of water, oxygen, and nutritious biomass. Additionally, the space vehicles must be well equipped to deal with metabolic human waste. It is therefore important to develop life-support systems which make these missions self-sufficient in terms of water, food, and oxygen production as well as waste management. One such solution is the employment of regenerative life-support systems that use biological and chemical/physical processes to recycle crew waste, revitalize air, and produce water and food. Photosynthetic cyanobacteria Limnospira could play a significant role in meeting these objectives. Limnospira can metabolize CO2 and nitrogen-rich human waste to produce oxygen and edible biomass. So far, life-support system studies have mainly focused on using chemical/physical methods to recycle water, degrade human waste, and recycle CO2 into oxygen. Nowadays, additional microbial processes are considered, such as nitrification of urea–ammonium–rich human waste and then using the nitrate for cyanobacterial cultivation and air vitalization. This cascade of multiple processes tends to increase the complexity of the life-support systems. The possibility of using non-nitrified urine for Limnospira cultivation can partially solve these issues. Our previous studies have shown that it is possible to cultivate Limnospira with urea and ammonium, the prominent nitrogen forms present in non-nitrified urine. In this study, we investigated the possibility of cultivating Limnospira with the different nitrogen forms present in non-nitrified urine and also evaluated their effect on the oxygen production capacity of Limnospira. For this 35-day-long study, we worked on a simplified version of the European Space Agency’s MELiSSA. During this ground demonstration study, we monitored the effect of urea and ammonium (vs. nitrate) on the oxygen production capacity of Limnospira. A deterministic control law, developed and validated on the basis of a stochastic light-transfer model, modulated (increase/decrease) the incident light on the photobioreactor (with Limnospira) to control oxygen levels in the closed loop. The CO2 from the mouse compartment was recycled as a carbon source for Limnospira. We observed that while the system could meet the desired oxygen levels of 20.3% under the nitrate and urea regime, it could only reach a maximum O2 level of 19.5% under the ammonium regime.

Use of Photobioreactors in Regenerative Life Support Systems for Human Space Exploration.

There are still many challenges to overcome for human space exploration beyond low Earth orbit (LEO) (e.g., to the Moon) and for long-term missions (e.g., to Mars). One of the biggest problems is the reliable air, water and food supply for the crew. Bioregenerative life support systems (BLSS) aim to overcome these challenges using bioreactors for waste treatment, air and water revitalization as well as food production. In this review we focus on the microbial photosynthetic bioprocess and photobioreactors in space, which allow removal of toxic carbon dioxide (CO2) and production of oxygen (O2) and edible biomass. This paper gives an overview of the conducted space experiments in LEO with photobioreactors and the precursor work (on ground and in space) for BLSS projects over the last 30 years. We discuss the different hardware approaches as well as the organisms tested for these bioreactors. Even though a lot of experiments showed successful biological air revitalization on ground, the transfer to the space environment is far from trivial. For example, gas-liquid transfer phenomena are different under microgravity conditions which inevitably can affect the cultivation process and the oxygen production. In this review, we also highlight the missing expertise in this research field to pave the way for future space photobioreactor development and we point to future experiments needed to master the challenge of a fully functional BLSS.

Optimizing Nitrogen Fixation and Recycling for Food Production in Regenerative Life Support Systems

Nitrogen (N) recycling is essential for efficient food production in regenerative life support systems. Crew members with a high workload need 90–100 g of protein per person per day, which is about 14 g of N, or 1 mole of N, per person per day. Most of this N is excreted through urine with 85% as urea. Plants take up N predominantly as nitrate and ammonium, but direct uptake as urea is possible in small amounts. Efficient N recycling requires maintenance of pH of waste streams below about 7 to minimize the volatilization of N to ammonia. In aerobic reactors, continuous aerobic conditions are needed to minimize production and volatilization of nitrous oxide. N is not well recycled on Earth. The energy intensive Haber–Bosh process supplies most of the N for crop production in terrestrial agriculture. Bacterial fixation of dinitrogen to ammonium is also energy intensive. Recycling of N from plant and human waste streams is necessary to minimize the need for N fixation. Here we review approaches and potential for N fixation and recycling in regenerative life support systems. Initial estimates indicate that nearly all the N from human and plant waste streams can be recovered in forms usable for plants.

ИЗУЧЕНИЕ ПОТРЕБЛЕНИЯ БЕЛЬЯ, ОДЕЖДЫ, СРЕДСТВ ЛИЧНОЙ ГИГИЕНЫ В ХОДЕ ЭКСПЕРИМЕНТОВ ПО ПРОЕКТУ SIRIUS

For the 17- and 120-day isolation campaigns within project SIRIUS, the stock of personal hygiene items (PHIs), underwear and clothes was supplied in compliance with the current ISS rates of consumption. Over 120 questionnaires filled by the human subjects made possible estimation of the total weight of these provisions in a mission to the Moon, and get feedback about the hygienic means and methods. The hygienic comfort of 6 crewmembers of the 120-d isolation campaign required more than 500 kg of underwear, bedclothes, towels, personal hygiene items, pieces of toilet and household essentials, which are also musts in a mission to the Moon and back. Long-duration missions and sojourns on lunar and Mars bases will call for designing arrangements for personal hygiene, shower, dish washing and laundry integrated with regenerative life support systems. Results of the investigation showed effectiveness of the sanitation and hygiene system and its functional significance.

Fundamental Study on Plant-based Regenerative Life-support Systems with Sweetpotato Culture in Space

Fundamental Study on Plant-based Regenerative Life-support Systems with Sweetpotato Culture in Space

Bio-electrochemical COD removal for energy-efficient, maximum and robust nitrogen recovery from urine through membrane aerated nitrification

Resource recovery from source-separated urine can shorten nutrient cycles on Earth and is essential in regenerative life support systems for deep-space exploration. In this study, a robust two-stage, energy-efficient, gravity-independent urine treatment system was developed to transform fresh real human urine into a stable nutrient solution. In the first stage, up to 85% of the COD was removed in a microbial electrolysis cell (MEC), converting part of the energy in organic compounds (27–46%) into hydrogen gas and enabling full nitrogen recovery by preventing nitrogen losses through denitrification in the second stage. Besides COD removal, all urea was hydrolysed in the MEC, resulting in a stream rich in ammoniacal nitrogen and alkalinity, and low in COD. This stream was fed into a membrane-aerated biofilm reactor (MABR) in order to convert the volatile and toxic ammoniacal nitrogen to non-volatile nitrate by nitrification. Bio-electrochemical pre-treatment allowed to recover all nitrogen as nitrate in the MABR at a bulk-phase dissolved oxygen level below 0.1 mg O2 L−1. In contrast, feeding the MABR directly with raw urine (omitting the first stage), at the same nitrogen loading rate, resulted in nitrogen loss (18%) due to denitrification. The MEC and MABR were characterised by very distinct and diverse microbial communities. While (strictly) anaerobic genera, such as Geobacter (electroactive bacteria), Thiopseudomonas, a Lentimicrobiaceae member, Alcaligenes and Proteiniphilum prevailed in the MEC, the MABR was dominated by aerobic genera, including Nitrosomonas (a known ammonium oxidiser), Moheibacter and Gordonia. The two-stage approach yielded a stable nitrate-rich, COD-low nutrient solution, suitable for plant and microalgae cultivation.

О функциональной надежности регенерационных систем жизнеобеспечения пилотируемых космических аппаратов

The paper considers two classes of problems concerned with reliability of manned spacecraft regenerative life-support system (RLSS) on the stages of design, testing and operation, as well as the approach to the definition of “functional reliability” as a component of the general reliability theory as opposed to the “structural reliability”. General methods for calculating the reliability of technological systems at all stages of their life cycle are considered. Mathematical models of functional, structural, and overall reliability of a manned spacecraft RLSS are proposed. Based on the results of independent tests of the carbon dioxide processing system, calculations of the functional reliability of the unit for the hydrogenation of carbon dioxide are performed. The unit is being developed for use in the manned spacecraft RLSS. The article also presents the conclusions on the analysis of the results of the calculation of functional reliability for the carbon dioxide hydrogenation unit.

Performance Evaluation of the Regenerative Life Support System of Interplanetary Manned Spacecraft.

The paper considers basic requirements for the efficiency model of the regenerative life-support system as an abiotic part of the ecological and technical system of an interplanetary spacecraft. It also describes developed efficiency model, which includes a set of indicators for solving the initial task of synthesis of the technological structure.

Surfactants effect on aeroponics and important mass balances of regenerative life support system – Lettuce case study

In order to be able to permanently settle other celestial bodies, it is necessary to create an efficient and closed life support system. Such a system will allow high autonomy and significantly reduce operating costs in a future colony. Grey water is a major waste stream in terms of the water volume generated by colonists, and its reuse is necessary. One possibility is to reuse it in aeroponic cultivation without treatment, which will in turn reduce its transported mass. The article focuses on water, carbon and other elements (N, P, K, Ca, Mg, Na) that are included in the mass balance of part of a life support system containing the aeroponic cultivation of lettuce that is supplied with clean water and water contaminated with one surfactant, crewmembers and a urine nitrification reactor. Three surfactants (Sodium Laureth Sulfate (SLES), Sodium Dodecylbenzene Sulfonate (SDBS), Sodium Methyl Cocoyl Taurate (SMCT)) in two concentrations, reflecting concentrated and diluted grey water, were tested. The growth of lettuce exposed to low concentrations (ca. 0.07 g·L−1) of surfactants has a moderate (SLES, SMCT) or no (SDBS) impact on the mass of grown plant. Exposition to high concentrations (>1.0 g·L−1) led to the complete failure of cultivation (SDBS, SMCT) or to very limited growth (SLES). In all cases, exposition to surfactants in low concentrations causes differences in the mineral composition of lettuce. In most cases, crops cultivated on water containing surfactants were characterized by a decreased harvest index (the ratio of edible biomass to total biomass). For the most cultivations the relative mass balances of water, C, N, P and K were generally unaffected by surfactants, while for Ca, Mg and Na, noticeable differences occurred. The data provided in the paper gives clues about which surfactant could be used in a future extraterrestrial colony.

Assessment of carbon recovery from solid organic wastes by supercritical water oxidation for a regenerative life support system

The carbon recovery from organic space waste by supercritical water oxidation (SCWO) was studied to support resource recovery in a regenerative life support system. Resource recovery is of utmost importance in such systems which only have a limited total amount of mass. However, the practical waste treatment strategies for solid space wastes employed today are only storing and disposal without further recovery. This work assesses the performance of SCWO at recovering organic wastes as CO2 and water, to discuss the superiority of SCWO over most present strategies, and to evaluate the different SCWO reactor systems for space application. Experiments were carried out with a batch and a continuous reactor at different reaction conditions. The liquid and gas products distribution were analyzed to understand the conversion of organics in SCWO. Up to 97% and 93% of the feed carbon were recovered as CO2 in the continuous and the batch reactor, respectively. Residual carbon was mostly found as soluble organics in the effluent. Compared with the batch reactor, the continuous reactor system demonstrated a ten times higher capacity within the same reactor volume, while the batch reactor system was capable of handling feeds that contained particulate matter though suffering from poor heat integration (hence low-energy efficiency) and inter-batch variability. It was concluded that SCWO could be a promising technology to treat solid wastes for space applications. A continuous reactor would be more suitable for a regenerative life support system.

Применение гидратцеллюлозной пленки для исследования роста и развития корневой системы модельного растения Arabidopsis thaliana L.

A new technique for Arabidopsis thaliana cultivation has been proposed that combines the use of a phytogel-based nutrient medium and a hydrophilic membrane of hydrate cellulose film, separating the root system of the plant from the medium thickness. Growth rates of both main and lateral roots were faster in the plants cultivated on the surface of hydrate cellulose film than in the plants grown in the phytogel volume. The location of the root system on the surface of the transparent hydrate film simplifies its observation and analysis and facilitates plant transplantation with preservation of the root system configuration. The proposed technique allowed us to first assess the effect of exogenous auxin on the growth of lateral roots at the 5-6 developmental stage. methods to study plant root systems, hydrate cellulose film, A. thaliana, lateral roots, differential root growth rate, auxin The work was financially supported by the Russian Foundation for Basic Research (Project Bel_mol_a 19-54-04015) and the basic topic of the Russian Academy of Sciences - IBMP RAS «Regularities of the Influence of Extreme Environmental Factors on the Processes of Cultivation of Higher Plants and the Development of Japanese Quail Tissues at Different Stages of its Ontogenesis under the Conditions of Regenerative Life Support Systems».

Eu:CROPIS AIV Program: Challenges and Solutions for a Spin-Stabilized Satellite Containing Biology

Eu:CROPIS is DLR’s first mission of the Compact Satellite Program. Its primary payload focuses on the research of closed-loop biological, regenerative life support systems, in a simulated gravitational environment of the Moon and Mars over months at a time. This is achieved by rotation of the satellite around its central body axis, using only magnetic torquers as actuators. A secondary payload (“PowerCells”) by the NASA Ames Research Center also utilizes the artificial gravity to conduct growth experiments on genetically modified organisms (GMOs). These payloads and the system design imposed constraints which affected the Assembly Integration and Verification (AIV) program in various ways and created challenges for the relatively small team to find solutions for. The paper to be presented will address the different aspects of the AIV program. This includes the verification of different critical components like the newly developed CFRP pressure vessel containing the primary payload and the Micrometeoroid and Debris Protection Shield, which protects it. Both items went through rigorous testing, including high-velocity impact tests, to ensure their reliability in orbit. Various other aspects concerning the biology had to be taken into account during AIV campaigns: due to the presence of degradable components within the primary payload, a late access capability had to be implemented in order to exchange biology as well as chemistry in cases of launch delays. To allow these operations as close as six months prior to launch, a highly flexible and streamlined acceptance test campaign was developed. A major impact on test planning and logistics was the fact that the secondary payload “PowerCells” contains GMOs, which European and German regulations restrict to be handled exclusively in especially certified laboratories (biosafety level 1 (BSL-1)). Thus, the use of external test facilities for the flight model campaign was not feasible as no European test center is certified to BSL-1. In consequence, the clean room facilities of the DLR Institute of Space Systems had to be certified to BSL-1 and new test infrastructure had to be procured in a short time frame to cover for acceptance testing. The design of the satellite and nature of the attitude control subsystem required limits on the magnetic momentum of the system and every unit it contains. A test flow incorporating the magnetic property measurement of each unit and a final system-level test in an external facility had to be devised, which enabled budgeting and projection of expected measurement results on the system level. Furthermore, the moments of inertia had to be measured precisely in order to have a stable spinning axis enabling a stable gravity simulation. Finally, the functionality had to be verified for each unit and for the system which required that several small test campaigns had to be conducted, like a solar panel deployment test and extensive software testing. A tight link to the operations teams of the German Space Operations and Control Center during such tests and beyond finally ensures the operability of the overall system in the operational phase.