Zebrafish cultured scales have been used effectively to study cellular and molecular responses of bone cells. In order to expose zebrafish scales to simulated microgravity (SMG) and/or vibration, we first determined via apoptosis staining whether cells of the scale survive in culture for two days and hence, we restricted our analyses to two-day durations. Next, we measured the effects of SMG and vibration on cell death, osteoclast tartrate-resistant acid phosphatase, and osteoblast alkaline phosphatase activity and on the number of Runx2a positive cells. We found that during the SMG treatment, osteoclast tartrate-resistant acid phosphatase activity increased on average, while the number of Runx2a positive cells decreased significantly. In contrast, SMG exposure caused a decrease in osteoblast activity. The vibration treatment showed an increase, on average, in the osteoblast alkaline phosphatase activity. This study demonstrates the effect of SMG and vibration on zebrafish scales and the effects of SMG on bone cells. We also show that zebrafish scales can be used to examine the effects of SMG on bone maintenance.
As part of planning for future space exploration, COSPAR (The Committee on Space Research) together with participating space agencies, organized and held interdisciplinary meetings to consider next steps in addressing knowledge gaps for planetary protection for future human missions to Mars. Beginning with the results of these meetings and earlier work by NASA, ESA, and COSPAR (e.g., Criswell et al., 2005; Hogan et al., 2006; Rummel et al., 2008) as a base the authors of this paper carried out a follow-on NASA planning activity to identify the necessary steps to be accomplished to close knowledge gaps. We identified significant overlap between the planetary protection needs and other sets of Mars preparation roadmaps (1) microbial monitoring requirements for crew health and medical systems, (2) studies of the microbiome of the built environment, (3) environmental control and life support systems (ECLSS), (4) waste management, and (5) planetary surface operations. In many cases, efforts to mature exploration class systems for Mars that are occurring in other domains can be leveraged with minor changes to address planetary protection gaps as well. In other cases, work planned for testing on the International Space Station (ISS) as an analog for crew Mars transit, or on the lunar surface as an analog for Mars surface operations can be used to close planetary protection technology and knowledge gaps. An overall strategic framework that combines these domains has the advantage of being more comprehensive, efficient, and timely for closing gaps.This approach has led to the development of a NASA roadmap for addressing planetary protection integrated with other related roadmaps. NASA's development and execution of the planetary protection is now viewed in an integrated way with related technology development and testing. Key features of the integrated capabilities roadmap include:center dot A focus on microbial monitoring needs for crewed systems overall, and the additional work needed to develop and demonstrate baseline vs. abnormal changes in microbial communities over time. center dot Integrated consideration of planetary protection and crew health requirements relevant to crew quarantine and science sample transportation.center dot Specific and targeted studies of microbial growth and biofilms in microgravity and lunar surface ECLSS systems.center dot Development of sterilization, cleaning, and waste handling technologies that meet habitation needs as well as planetary protection needs.
Understanding the structural and antibiotic resistance changes of microbial communities in space environments is critical for identifying potential pathogens that may pose health risks to astronauts and for preventing and controlling microbial contamination. The research to date on microbes under simulated space factors has primarily been carried out on single bacterial species under the individual effects of microgravity or low-dose radiation. However, microgravity (MG) and low-dose ionizing radiation (LDIR) coexist in the actual spacecraft environment, and microorganisms coexist as communities in the spacecraft environment. Thus, the microbial response to the real changes present during space habitation has not been adequately explored. To address this knowledge gap, we compared the dynamics of community composition and antibiotic resistance of synthetic bacterial communities under simulated microgravit, low-dose ionizing radiation, and the conditions combined, as it occurs in spacecraft. To ensure representative bacteria were selected, we co-cultured of 12 bacterial strains isolated from spacecraft cleanrooms. We found that the weakened competition between communities increased the possibility of species coexistence, community diversity, and homogeneity. The number of Bacilli increased significantly, while different species under the combined conditions showed various changes in abundance compared to those under the individual conditions. The resistance of the synthetic community to penicillins increased significantly under low doses of ionizing radiation but did not change significantly under simulated microgravity or the combined conditions. The results of functional predictions revealed that antibiotic biosynthesis and resistance increased dramatically in the community under space environmental stress, which confirmed the results of the drug sensitivity assays. Our results show that combined space environmental factors exert different effects on the microbial community structure and antibiotic resistance, which provides new insights into our understanding of the mechanisms of evolution of microorganisms in spacecraft, and is relevant to effective microbial pollution prevention and control strategies.
Under the initial stage of muscle mechanical unloading, the skeletal muscle undergo accumulation of high-energy phosphates followed by AMP-dependent proteinkinase (AMPK) inactivation. Since AMPK is known to activate mitochondrial biogenesis, it cannot be excluded that AMPK inactivation results in oxidative potential decrease at the later stages of muscle unloading. We decided to test the role of the accumulation of high-energy phosphates in skeletal muscle fibers in the inactivation of mitochondrial biogenesis regulators at an early stage of muscle unloading. To reduce the ATP/ADP ratio, we used beta-guanidine propionic acid, and the obtained data indicating that already during the first day of simulated microgravity, the accumulation of high-energy phosphates can reduce the expression level of mRNA of the key regulator of mitochondrial biogenesis PGC-1α, the transcription factor TFAM, as well as the mitochondrial fusion regulator - mitofusin-1. A number of other parameters of mitochondrial signaling were not subject to changes at this time-point. Thus, we demonstrated the role of the ATP/ADP ratio in the inactivation of several regulators of mitochondrial biogenesis in the postural soleus muscle at an early stage of functional unloading.
Health effects of space radiation are a serious concern for astronauts on long-duration missions. The lens of the eye is one of the most radiosensitive tissues in the body and, therefore, ocular health risks for astronauts is a significant concern. Studies in humans and animals indicate that ionizing radiation exposure to the eye produces characteristic lens changes, termed "radiation cataract," that can affect visual function. Animal models of radiation cataractogenesis have previously utilized inbred mouse or rat strains. These studies were essential for determining morphological changes and dose-response relationships between radiation exposure and cataract. However, the relevance of these studies to human radiosensitivity is limited by the narrow phenotypic range of genetically homogeneous animal models. To model radiation cataract in genetically diverse populations, longitudinal cataract phenotyping was nested within a lifetime carcinogenesis study in male and female heterogeneous stock (HS/Npt) mice exposed to 0.4 Gy HZE ions (n = 609) or 3.0 Gy gamma-rays (n = 602) and in unirradiated controls (n = 603). Cataractous change was quantified in each eye for up to 2 years using Merriam-Focht grading criteria by dilated slit lamp examination. Virtual Optomotry (TM) measurement of visual acuity and contrast sensitivity was utilized to assess visual function in a subgroup of mice. Prevalence and severity of posterior lens opacifications were 2.6-fold higher in HZE ion and 2.3-fold higher in gamma-ray irradiated mice compared to unirradiated controls. Male mice were at greater risk for spontaneous and radiation associated cataracts. Risk for cataractogenesis was associated with family structure, demonstrating that HS/Npt mice are well-suited to evaluate genetic determinants of ocular radiosensitivity. Last, mice were extensively evaluated for cataract and tumor formation, which revealed an overlap between individual susceptibility to both cancer and cataract.
Monitoring space radiation is of vital importance for risk reduction strategies in human space exploration. Radiation protection programs on Earth and in space rely on personal and area radiation monitoring instruments. Crew worn radiation detectors are crucial for successful crew radiation protection programs since they measure what each crewmember experiences in different shielding configurations within the space habitable volume. The Space Radiation Analysis Group at NASA Johnson Space Center investigated several compact, low power, real-time instruments for personal dosimetry. Following these feasibility studies, the Crew Active Dosimeter (CAD) has been chosen as a replacement for the legacy crew passive radiation detectors. The CAD device, based on direct ion storage technology, was developed by Mirion Dosimetry Services to meet the specified NASA design requirements for the International Space Station (ISS) and Artemis programs. After a successful Technology demonstration on ISS, the CAD has been implemented for ISS Crew operations since 2020. The current paper provides an overview of the CAD development, ISS results and comparison with the ISS Radiation Assessment Detector (RAD) and the Radiation Environment Monitor 2 (REM2) instruments.
Modern pupillometers are automated, thereby providing an objective, accurate, and reliable evaluation of various aspects of the pupillary light reflex at precision levels that were previously unobtainable. There are many gaps in knowledge regarding pupil size and pupillary light reflex in nervous system changes related to space travel given the previous lack of a precise method to quantitatively measure it. Automated pupillometry has not been used previously in space. This novel tool has promising uses in altered gravity environments as a sensitive non-invasive tool to determine alterations due to headward fluid shifts and elevated intracranial pressure. This article discusses the potential use of automated pupillometry in space for monitoring of astronaut health and neurological pathology.
The response of plants to radiation is an essential topic in both space plant cultivation and mutation breeding by radiation. In this study, heavy ion beams (HIB) generated by the ground accelerator and X-rays (XR) were used as models of high linear energy transfer (LET) and low LET radiation to study the molecular response mechanism of Platycodon grandiflorus (P. grandiflorus) seedlings after irradiation. The gene and protein expression profiles of P. grandiflorus after 15 Gy HIB and 20 Gy XR radiation were analyzed by transcriptome and proteome. The results showed that the number of differentially expressed genes (DEGs) induced by HIB radiation was less than that of XR group, but HIB radiation induced more differentially expressed proteins (DEPs). Both HIB and XR radiation activated genes of RNA silencing, double-strand break repair and cell catabolic process. DNA replication and cell cycle related genes were down-regulated. The genes of cell wall and external encapsulating structure were upregulated after HIB radiation. The gene expression of protein folding and glucan biosynthesis increased after XR radiation. Protein enrichment analysis indicated that HIB radiation resulted in differential protein enriched in photosynthesis and secondary metabolite biosynthesis pathways, while XR radiation induced differential protein of glyoxylate and dicarboxylate metabolism and carbon metabolism. After HIB and XR radiation, the genes of antioxidant system and terpenoid and polyketide metabolic pathways presented different expression patterns. HIB radiation led to the enrichment of non-homologous end-joining pathway. The results will contribute to understanding the biological effects of plants under space radiation.
Humans have stepped on the Lunar surface for less than 80 h of Extravehicular Activity, providing a narrow understanding of Lunar gait patterns. NASA's Human-crewed Artemis missions are quickly approaching; un-derstanding how fractional gravity affects gait patterns will be critical for the Moon's and Mars' long-term habitation. This study examined gait patterns under 1.0 g (Earth), simulated 0.38 g (Martian), and 0.17 g (Lunar). Participants walked and ran on a treadmill supported by ARGOS (Active Response Gravity Offload System), simulating fractional gravity. Vicon motion capture data and principal component analysis software were used to capture and quantify coordinated gait structures. There were found to be significant differences (p < 0.05) in the coordinative gait structures for ambulation between fractional gravity conditions. Additionally, there were significantly higher asymmetric gait components for Lunar conditions. Finally, a skipping coordi-native structure was identified within Lunar and Martian running.
Sterols are the main components of the plasma membrane and are involved in various plant membrane functions. Azuki bean (Vigna angularis (Wild.) Ohwi et Ohashi) seedlings were cultivated under hypergravity conditions, and changes in the levels and composition of membrane sterols in their epicotyls were analyzed. Under hypergravity conditions at 300 g, the levels of steryl glycosides and acyl steryl glycosides per unit length and per gram fresh weight greatly increased, which accounted for an increase in the total sterol levels. Stigmasterol, & beta;-sitosterol, and campesterol were the most abundant sterols. Hypergravity decreased the proportion of stig-masterol but increased that of & beta;-sitosterol. The fatty chains of acyl steryl glycosides mainly consisted of palmitic acid (C16:0), stearic acid (C18:0), linoleic acid (C18:2), and & alpha;-linolenic acid (C18:3), and their proportions were not modified under hypergravity conditions. In addition, the density of membrane microdomains, visualized with anti-Flotillin 1 antibody per unit area, increased by hypergravity, suggesting that lipid raft formation was stimulated. These results support the hypothesis that lipid rafts are involved in plant response and resistance to gravity.
We detect regular particle showers in several compact pixel detectors, distributed over the International Space Station. These showers are caused by high energy galactic cosmic rays, with energies often in the 10 s of TeV or higher. We survey the frequency of these events, their dependence on location on ISS, and their independence of the location of ISS, on its orbit. The Timepix detectors used allow individual particle tracks to be resolved, providing a possibility to perform physical analysis of shower events, which we demonstrate. In terms of radiation dosimetry, these showers indicate certain possible limitations of traditional dosimetric measures, in that (a) the dose measured in small sensor may be less than that received in a larger distribution of matter, such as a human and (b) the spatial and temporal extent of these events represents a regime of poorly documented biological response.
Space travel and exploration are associated with increased ambient CO2 (i.e., a hypercapnic environment). Some work reported that the physiological changes (e.g., increased cerebral blood flow [CBF]) associated with a chronic hypercapnic environment contributes to a "space fog" that adversely impacts cognition and psychomotor performance, whereas other work reported no change or a positive change. Here, we employed the antisaccade task to evaluate whether transient exposure to a hypercapnic environment influences top -down executive function (EF). Antisaccades require a goal -directed eye movement mirror -symmetrical to a target and are an ideal tool for identifying subtle EF changes. Healthy young adults (aged 19-25 years) performed blocks of antisaccade trials prior to (i.e., pre -intervention), during (i.e., concurrent) and after (i.e., post -intervention) 10 -min of breathing factional inspired CO2 (FiCO2) of 2.5% (i.e., hypercapnic condition) and during a normocapnic (i.e., control) condition. In both conditions, CBF, ventilatory and cardiorespiratory responses were measured. Results showed that the hypercapnic condition increased CBF, ventilation and end -tidal CO2 and thus demonstrated an expected physiological adaptation to increased FiCO2. Notably, however, null hypothesis and equivalence tests indicated that concurrent and post -intervention antisaccade reaction times were refractory to the hypercapnic environment; that is, transient exposure to a FiCO2 of 2.5% did not produce a real-time or lingering influence on an oculomotor-based measure of EF. Accordingly, results provide a framework that - in part - establishes the FiCO2 percentage and timeline by which high-level EF can be maintained. Future work will explore CBF and EF dynamics during chronic hypercapnic exposure as more direct proxy for the challenges of space flight and exploration.
During space travel, the gut microbiota is changed which can lead to health-related issues. Previously, we utilized the hind-limb unloaded (HU) mouse, which is an established ground-based in-vivo model of microgravity and observed altered gut microbiota. In this study, we evaluated the beneficial effects of novel bacterial conditioned media in HU mice to understand if they can offset the effects of unloading in the HU mouse model. We aimed to explore the influence of bacterial conditioned media on diversity and quantity of intestinal microbes in HU mice, and investigated the microarchitecture of mice retinas and kidneys to evaluate the potential systemic effects of bacterial conditioned media in HU mice. Four-month-old, male C57/Bl6 mice were separated into groups: including the ground-based control group, the HU group mice fed with vehicle as placebo (HU-placebo mice), and the HU group fed with bacterial conditioned media (HU-CP mice) and kept under controlled environmental conditions for three weeks. Next, mice were sacrificed; gut dissections were conducted, and metagenomic analysis of bacterial species was performed via DNA extraction and 16S rRNA analysis. The results revealed an HU-induced reduction in intestinal microbial diversity, and an increase in pathogenic bacteria dominated by Firmicutes (45%). In contrast, supplementation with bacterial conditioned media for three weeks led to a significant increase in gut microbial diversity with noticeable changes in the OTUs abundance in the HU mice. Additionally, HU-induced muscle weakness and structural abnormalities in the retina and kidney were partially prevented with bacterial conditioned media. Moreover, a greater diversity of several bacteria in the HU-CP was observed including, Bacteriodota, Firmicutes, Proteobacteria, Actionobacteriota, Verrucomicorbiota, Cyanobacteria, Gemmatimonadota, Acidobacteriota, Chloroflexi, Myxococcota, and others. Prospective research involving molecular mechanistic studies are needed to comprehend the systemic effects of bacterial metabolites conditioned media on experimental animal models under chronic stress.
Purpose: Astronauts on missions beyond low Earth orbit will be exposed to galactic cosmic radiation, and there is concern about potential adverse cardiovascular effects. Most of the research to identify cardiovascular risk of space radiation has been performed in rodent models. To aid in the translation of research results to humans, the current study identified long-term effects of high-energy charged particle irradiation on cardiovascular function and structure in a larger non-rodent animal model.Materials and methods: At the age of 12 months, male New Zealand white rabbits were exposed to whole-body protons (250 MeV) or oxygen ions (16O, 600 MeV/n) at a dose of 0 or 0.5 Gy and were followed for 12 months after irradiation. Ultrasonography was used to measure in vivo cardiac function and blood flow parameters at 10- and 12-months post-irradiation. At 12 months after irradiation, blood cell counts and blood chemistry values were assessed, and cardiac tissue and aorta were collected for histological as well as molecular and biochemical analyses. Plasma was used for metabolomic analysis and to quantify common markers of cardiac injury.Results: A small but significant decrease in the percentage of circulating lymphocytes and an increase in neutrophil percentage was seen 12 months after 0.5 Gy protons, while 16O exposure resulted in an increase in monocyte percentage. Markers of cardiac injury, cardiac troponin I (cTnI) and N-Terminal pro-B-type Natriuretic Peptide were modestly increased in the proton group, and cTnI was also increased after 16O. On the other hand, metabolomics on plasma at 12 months revealed no changes. Both types of irradiation demonstrated alterations in cardiac mitochondrial morphology and an increase in left ventricular protein levels of inflammatory cell marker CD68. However, changes in cardiac function were only mild.Conclusion: Low dose charged particle irradiation caused mild long-term changes in inflammatory markers, cardiac function, and structure in the rabbit heart, in line with previous studies in mouse and rat models.
In view of the critical role the gut microbiome plays in human health, it has become clear that astronauts' gut microbiota composition changes after spending time in space. Astronauts are exposed to several risks in space, including a protracted period of microgravity, radiation, and mechanical unloading of the body. Several deleterious effects of such an environment are reported, including orthostatic intolerance, cardiovascular endothelial dysfunction, cellular and molecular changes, and changes in the composition of the gut microbiome. Herein, the correlation between the gut microbiome and cardiovascular disease in a microgravity environment is evaluated. Additionally, the relationship between orthostatic hypotension, cardiac shrinkage and arrhythmias during spaceflight, and cellular alterations during spaceflight is reviewed. Given its impact on human health in general, modifying the gut microbiota may significantly promote astronaut health and performance. This is merited, given the prospect of augmented human activities in future space missions.
The National Aeronautics and Space Administration (NASA) has rigorously documented a group of neuro-ophthalmic findings in astronauts during and after long-duration spaceflight known as spaceflight associated neuro-ocular syndrome (SANS). For astronaut safety and mission effectiveness, understanding SANS and countermeasure development are of utmost importance. Although the pathogenesis of SANS is not well defined, a leading hypothesis is that SANS might relate to a sub-clinical increased intracranial pressure (ICP) from cephalad fluid shifts in microgravity. However, no direct ICP measurements are available during spaceflight. To further understand the role of ICP in SANS, pupillometry can serve as a promising non-invasive biomarker for spaceflight environment as ICP is correlated with the pupil variables under illumination. Extended reality (XR) can help to address certain limitations in current methods for efficient pupil testing during spaceflight. We designed a protocol to quantify parameters of pupil reactivity in XR with an equivalent time duration of illumination on each eye compared to pre-existing, non-XR methods. Throughout the assessment, the pupil diameter data was collected using HTC Vive Pro-VR headset, thanks to its eye-tracking capabilities. Finally, the data was used to compute several pupil variables. We applied our methods to 36 control subjects. Pupil variables such as maximum and minimum pupil size, constriction amplitude, average constriction amplitude, maximum constriction velocity, latency and dilation velocity were computed for each control data. We compared our methods of calculation of pupil variables with the non-XR methods existing in the literature. Distributions of the pupil variables such as latency, constriction amplitude, and velocity of 36 control data displayed near-identical results from the non-XR literature for normal subjects. We propose a new method to evaluate pupil reactivity with XR technology to further understand ICP's role in SANS and provide further insight into SANS countermeasure development for future spaceflight.
Space foods closely associate with the performance and mental health of astronauts. Over the years, a range of manufacturing technologies have been explored and advancements in food 3D printing can provide answers to certain existing challenges and revolutionize the way foods are prepared for space exploration missions. Apart from the nutrition and satiety perspective, product shelf-life, variety, personalization, and the need for customized diets are critical considerations. In such long-duration human-crewed space missions, under microgravity conditions and exposure to space, psychological factors heavily affect food consumption patterns. Therefore, there has been a surge in research funding for developing products and methods that offer safe, nutritionally balanced, and delightful food options. 3D food printing could be a creative solution for such requirements. While multiple challenges must be addressed, the technology promises waste minimization and the scope for on-site on-demand food preparation. This article begins with fundamental concepts of this subject, provides a timeline of the advancements in the field, and details the futuristic prospects of the technology for long-duration space missions.
Female germ cells provide the structural basis for the development of a new organism, while the main molecular mechanisms of the impact of weightlessness on the cell remain unknown. The aim of this work was to determine the relative content and distribution of the main proteins of microtubules and microfilaments, to assess the relative RNA content of genes in mouse oocytes after short-term exposure to simulated microgravity, and to determine the potential for embryo development up to the 3-cell stage. Before starting the study, BALB/c mice were divided into two groups. One group received water and standard food without any modifications. Before exposure to simulated microgravity, the oocytes of these animals were randomly divided into two groups – c and µg. The second group of animals additionally received essential phospholipids containing at least 80% phosphatidylcholines, per os for 6 weeks before the start of the experiment at a dosage of 350 mg/kg of the animal's body to modify the lipid composition of the oocyte membrane. The obtained oocytes of these animals were also randomly divided into two groups – ce and µge. To determine the protein distribution and its relative content, immunofluorescence analysis was performed, and the RNA content of genes was assessed using real-time PCR with reverse transcription. After cultivation under simulated microgravity, beta-actin and acetylated alpha-tubulin are redistributed from the cortical layer to the central part of the oocyte, and the relative content of acetylated alpha-tubulin and tubulin isoforms decreases. At the same time, the mRNA content of most genes encoding cytoskeletal proteins was significantly higher in comparison with the control level. The use of essential phospholipids led to a decrease in the content of cellular cholesterol in the oocyte and leveled changes in the content and redistribution of acetylated alpha-tubulin and beta-actin after cultivation under simulated microgravity. In addition, after in vitro fertilization and further cultivation under simulated weightlessness, we observed a decrease in the number of embryos that passed the stage of the 2-cell embryo, but while taking essential phospholipids, the number of embryos that reached the 3-cell stage did not differ from the control group. The results obtained show changes in the content and redistribution of cytoskeletal proteins in the oocyte, which may be involved in the process of pronucleus migration, the formation of the fission spindle and the contractile ring under simulated weightlessness, which may be important for normal fertilization and cleavage of the future embryo.
Sleep deprivation (SD) is common during spaceflight. SD is known to cause cognitive deficits and depression, requiring treatment and prevention. Hemerocallis citrina Baroni (Liliaceae) is a perennial herb with antidepressant, antioxidant, antitumor, anti-inflammatory, and neuroprotective effects.The aim of our study was to investigate the effects of H. citrina extract (HCE) on SD-induced cognitive decline and depression-like behavior and possible neuroinflammation-related mechanisms. HCE (2 g/kg/day, i.g.) or vortioxetine (10 mg/kg/day, i.g.) were given to mice by oral gavage for a total of 28 days during the SD process. HCE treatment was found to ameliorate SD-induced impairment of short- and long-term spatial and nonspatial memory, measured using Y-maze, object recognition, and Morris water maze tests, as well as mitigating SD-induced depression-like behaviors, measured by tail suspension and forced swimming tests. HCE also reduced the levels of inflammatory cytokines (IL-1β, IL-18, and IL-6) in the serum and hippocampus. Furthermore, HCE suppressed SD-induced microglial activation in the prefrontal cortex (PFC) and the CA1 and dentate gyrus (DG) regions of the hippocampus. HCE also inhibited the expression of phosphorylated NF-κB and activation of the NLRP3 inflammasome. In summary, our findings indicated that HCE attenuated SD-induced cognitive impairment and depression-like behavior and that this effect may be mediated by the inhibition of inflammatory progression and microglial activation in the hippocampus, as well as the down-regulation of NF-κB and NLRP3 signaling. The findings of these studies showingTthese results indicate that HCE exerts neuroprotective effects and are consistent with the findings of previous studies, suggesting that HCE is beneficial for the prevention and treatment of cognitive decline and depression in SD.
Space agencies are developing Bioregenerative Life Support Systems (BLSS) in view of upcoming long-term crewed space missions. Most of these BLSS plan to include various crops to produce different types of foods, clean water, and O2 while capturing CO2 from the atmosphere. However, growing these plants will require the appropriate addition of nutrients in forms that are available. As shipping fertilizers from Earth would be too costly, it will be necessary to use waste-derived nutrients. Using the example of the MELiSSA (Micro-Ecological Life Support System Alternative) loop of the European Space Agency, this paper reviews what should be considered so that nutrients recycled from waste streams could be used by plants grown in a hydroponic system. Whereas substantial research has been conducted on nitrogen and phosphorus recovery from human urine, much work remains to be done on recovering nutrients from other liquid and solid organic waste. It is essential to continue to study ways to efficiently remove sodium and chloride from urine and other organic waste to prevent the spread of these elements to the rest of the MELiSSA loop. A full nitrogen balance at habitat level will have to be achieved; on one hand, sufficient N2 will be needed to maintain atmospheric pressure at a proper level and on the other, enough mineral nitrogen will have to be provided to the plants to ensure biomass production. From a plant nutrition point of view, we will need to evaluate whether the flux of nutrients reaching the hydroponic system will enable the production of nutrient solutions able to sustain a wide variety of crops. We will also have to assess the nutrient use efficiency of these crops and how that efficiency might be increased. Techniques and sensors will have to be developed to grow the plants, considering low levels or the total absence of gravity, the limited volume available to plant growth systems, variations in plant needs, the recycling of nutrient solutions, and eventually the ultimate disposal of waste that can no longer be used.
