Abstract EANA2026-43

The establishment of sustainable habitats on the Moon will require efficient Bioregenerative Life Support Systems (BLSS) capable of recycling waste and producing essential resources, ultimately minimizing dependence on Earth resupply. While cyanobacteria and plants have been extensively studied for their roles in such systems, fungi remain overlooked, despite their broad biotechnological potential—including the production of biocomposites for construction, melanin for radiation shielding, bioactive compounds, and applications in biomining. 

In this work, we present the SELENA project (Sustainable microbial Ecosystem for Lunar ExploratioN Applications), which aims to develop a proof-of-concept BLSS module integrating filamentous fungi into future lunar habitats. SELENA explores the use of (Aspergillus niger) using biomass from cyanobacteria and tomato plants (Solanum lycopersicum) as local nutrient sources. The project has three main objectives: a) select and characterize cyanobacterial strains capable of growing on Lunar regolith simulants under microgravity and Lunar gravity; b) identify the best medium composition using locally available resources to support A. niger growth; and c) establish a BLSS module prototype that uses a “fungi-cyanobacteria-plants” loop capable of quasi-steady-state operation.

Results demonstrate the feasibility of growing filamentous fungi using exclusively space-relevant resources. A. niger was successfully cultivated in liquid culture, at room temperature, using media composed of a) lysates of the cyanobacterium Anabaena sp. PCC 7938 at a concentration of 1g/L—a strain already established as a model organism for in situ resource utilization on Mars and provided through collaboration with Dr. Cyprien Verseux; and b) filtered aqueous extracts prepared from the lunar regolith simulant LHS-1 (Exolith) at 40 g/L. Under these conditions, A. niger reached approximately 30% of the biomass yield obtained in control conditions (standard minimal medium).

Additionally, using the same medium composition, we further validated A. niger growth  in Fluorinated Ethylene Propylene (FEP) gas-permeable bag-bioreactors (C-bags, Saint Gobain)—space-proven culture systems, previously flown aboard the ISS and Artemis I—under simulated microgravity through clinorotation. No statistical differences compared to Earth gravity controls were observed. These findings emphasize the feasibility of integrating filamentous fungi growth using only two likely available resources in a Moon mission scenario—cyanobacteria biomass and lunar minerals—and further validate FEP bags as a promising cultivation platform for filamentous fungi in space biotechnology applications.

Current work focuses on testing additional cyanobacterial strains for growth in aqueous extracts prepared from LHS-1 at different concentrations (40 g/L to 200 g/L), supplemented with 5 mM urea for non-heterocyst-forming strains. Strains under investigation include Synechocystis sp. PCC 6803, Gloeocapsopsis cf. crepidinum LEGE 06123/LEAN 026, Arthrospira platensis, Oculatella lusitanica LEGE 161147, Nostoc punctiforme PCC 73102 and Anabaena sp. PCC 7120. This research aims to identify the most robust and resource-efficient strain(s) for integration into the SELENA loop. 

Future tests will address a full prototype integration of fungi, cyanobacteria, and tomato plants into an operational BLSS prototype. Ultimately, the SELENA project will contribute to a greater autonomy and sustainability of future crewed missions to the Moon, by promoting closed-loop technologies which ultimately reduce reliance on Earth-derived supplies.