Pseudomonas fluorescens is classified as a biosafety level 1 microorganism, meaning that it is low-risk and poses little to no threat of infection in healthy adults 1 . It is an opportunistic pathogen with some strains infecting humans with compromised immunity.
Gene |
Organism of origin |
|---|---|
| crnA | Pseudomonas putida |
| creA | Pseudomonas putida |
| soxA | Bacillus subtilis |
| wspF | Especially designed for P. fluorescens and P. putida |
| nxrA | Nitrobacter winogradskyi Nb-255 (ATCC 25391) |
| nxrX | Nitrobacter winogradskyi Nb-255 (ATCC 25391) |
| nxrB | Nitrobacter winogradskyi Nb-255 (ATCC 25391) |
| amoC | Nitrosomonas europaea |
| amoA | Nitrosomonas europaea (ATCC 19718) |
| amoB | Nitrosomonas europaea (ATCC 19718) |
| hao | Nitrosomonas europaea (ATCC 19718) |
| rpoS | Pseudomonas fluorescens |
| hfQ | Pseudomonas fluorescens |
| KatB | Pseudomonas fluorescens |
All the genes used were safe (no toxin) and were either provided by IDT as synthetic DNA or by institutional researchers from our partner laboratories (TBI and TWB), or were directly amplified from genomic DNA.
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Training course on the proper use of the two autoclaves installed in our lab: we learnt about the technical principles of autoclaves, the different risks associated and how to use them. We were given authorization by the person in charge of their maintenance.
General hygiene and security training course required by TBI, the lab hosting us to efficiently understand important security aspects.
Quality training course required by TBI on the quality procedures at the lab. We learnt about different practices, the way the workforce is structured and the different people we could contact if we needed help or had any doubts.
etBR training course: since in our lab we have a special hazardous chemicals room dedicated to etBR, Christophe Danelon, our PI, gave us a training course on the proper use of the equipment inside the room (computer, printer, gel imager, etBR tank, etc.) and the safety procedures to follow (always throw away the gloves used inside in a dedicated trash can, never touch anything without gloves, etc.).
From the beginning of the project, BioMoon positioned itself in the context of an established lunar base, with the appropriate infrastructures to allow microbial and plant growth.
With bringing engineered bacteria extraterrestrially comes a series of risks which we have taken into consideration when planning the appropriate conditions for the application of our project. The engineered bacteria would allow plant growth inside lunar greenhouses.
The upkeep of these installations would be on the astronauts living there. The risk of BioMoon’s P.fluorescens strain being released beyond the greenhouse is minimal, mainly because it could not survive lunar conditions: radiation, extreme temperatures, and the lack of nitrogen and carbon. The appropriate use of our biostimulant on the Moon would need specific guidelines designed by experts on both plant growth and extraterrestrial bodies; they would need to ensure the safety of the users as well as maximal efficiency of our product.
Article 9 of the UN Outer Space Treaty of 1967 declares that: “States Parties to the Treaty shall pursue studies of outer space, including the Moon and other celestial bodies, and conduct exploration of them so as to avoid their harmful contamination and also adverse changes in the environment of the Earth resulting from the introduction of extraterrestrial matter, and where necessary, shall adopt appropriate measures for this purpose."
With this principle in mind, the Committee on Space Research (COSPAR) published, in 2021, its Policy on Planetary Protection, which was updated in 2024.
This document outlines the five different categories in which space missions and extraterrestrial bodies are separated and it gives clear guidelines on what the safety considerations should be, which our team has taken in great consideration.
BioMoon would be considered a Category 2 mission since it involves a Category 2 extraterrestrial body, the Moon.
“Category II missions comprise all types of missions to those target bodies where there is significant interest relative to the process of chemical evolution and the origin of life, but where there is only a remote chance that contamination carried by a spacecraft could compromise future investigations. The requirements are for simple documentation only.”
In compliance with COSPAR’s policy on Planetary Protection we would provide pre-mission documentation on the purpose of our technology as well as its possible impact. After this goal is accomplished, we would redact an End-of-Mission report detailing our impact, if there has been any.