EXPERIMENT RECORD N° 9214
CFS-A - Growth and Survival of Coloured Fungi in Space
  1. 2010 • ISS Increments 25-26
  2. 2010 • ISS "MagISStra"- long-duration mission
  3. 2011 • ISS Increments 27-28
Life Sciences:
  • Cell and Molecular Biology
Jason Hatton
jason.hatton@esa.int
D. Hasegan (1), I. Gomoiu (2), E. Chatzitheodoridis (3)
(1)  
University/Research Centre or SME: Romanian Institute of Space Science
Str. Atomistilor, Nr. 409
Jud Ilfov, Magurele
cod postal 077125
ROMANIA
Tel:  
+40(0)40214574471
Fax:  
+40(0)40214574471
e-mail:  
iss@venus.nipne.ro
(2)  
Institute of Biology
296 Splaiul Independentei
P.O. Box 56-53
Bucharest,060031
ROMANIA
Tel:  
+40(0)40212239072
Fax:  
+40(0)40212219071
e-mail:  
ioana.gomoiu@ibiol.ro
gomoiu@hotmail.com
(3)  
Director of the NTUA/Athens group
National Technical University of Athens
School of Mining and Metalurgical Engineering
Section of Geological Sciences
Athens
GREECE
AIM
Determine the effect of microgravity and cosmic radiation on the growth and survival of colored fungi species.

CFS-A experiment was performed in biocontainers fully integrated with Ulocladium chartarum colonies (0, 2 and 4 days old) in microcells and dried spore samples of Ulocladium chartarum, Aspergillus niger, Cladosporium herbarum and Basipetospora halophila on iron, silica and polycarbonate wafers put in microcapsules).

SPECIFIC GOALS

1. Measure colonial growth of Ulocladium sp. on agar during spaceflight.
The colonial growth of Ulocladium chartarum on agar nutrient medium, will be measured during spaceflight and in a ground control. The following parameters will be measured:
- diameters of colonies,
- length of active growth area,
- dynamic of sporulation using photography done during spaceflight.

2. Examine the survival and growth of different colored fungi species which can be relevant to contamination of spacecraft (e.g. ISS and other missions with crew).
Survival of species is measured post flight using colony forming ability of exposed spores.
Germination,
rate of growth,
sporulation and
new growth value parameters are established.

3. Determine if spaceflight conditions affect the sporulation of Ulocladium chartarum as a function of age of the colony.
Rate of sporulation is measured using Carnoy 2,0 software using pictures done during flight because spores are brown-dark. Post-flight optical microscopy done in the lab. Will check sporulation, spore color and morphology.

4. Observe the spreading & adhesion of Ulocladium chartarum spores on different substrates in microgravity.

5. High resolution imaging of spores and hyphae using optical microscopy and AFM to determine changes produced by cosmic radiation and microgravity as well as in normal conditions (lab).

6. Design of microcapsules and container for radiological research in space.

Note: This experiment is part A of the proposal. Part B of the proposal targets use of the EXPOSE facility to investigate survival of spores exposed to the external space environment, including Solar UV light.

The proposal “Growth and survival of colored fungi in space” consists of ground and space experiments both on the inside of microcapsules by researchers from Romanian Institute of Space Science. Microbiology activities and imaging of spores at Atomic Force Microscope will be done by a team from Romanian Institute of Biology and
National Hellenic Research Foundation, Athens.

The fungal species chosen for experiments belong to 4 genera selected as organic material decomposers, possible contaminants of materials destined for interplanetary travel, aggressive biodeteriogens of artworks and wooden buildings.

Those containing melanin are protected against UV rays. As mature cultures and thin film of spores they will be exposed in normal conditions on the Earth, in incubators with microgravity and cosmic radiation conditions and on the ISS where full conditions of space environmental are found. Black fungal spores survival in space is relevant to panspermia and planetary protection forward contamination. Data from these studies will be used for calculation of the survival and possible proliferation of potential contaminants transferred from Earth on spacecraft.

The project will give some results on whether life can be maintained and for how long in extreme condition existing in space (solar UV radiations, vacuum, cosmic radiation, temperature extreme).

Colonial growth of fungi, sporulation, spreading and adhesion of spores will be studied measuring rate of growth, intensity, optical microscopy. Survival value of fungal spores’ exposure to space and their germination are studied by optical microscopy, AFM and cultivation. Phenotypic and metabolic expressions of mutagenesis will also be characterized.

Colored fungi will be flown and allowed to develop in microgravity conditions. The effects of microgravity on development, metabolism and sporulation of the cultivated fungi will be measured.
Two sets of samples will be flown. One set containing live cultures and dried spores will aim at monitoring the growth, development and metabolism of fungi over a short duration mission in space. The second set will consist of dried spores only and will be stowed in space for at least 5 months to assess the combined impact of microgravity and radiation on sporulation.

Fungal species studied:
Ulocladium chartarum CM 1
Aspergillus niger CM 1
Cladosporium herbarum CM 1
Basipetospora halophila - CM 1

General Experiment Procedure:
Experiment team transports fungal cultures and dried spores to launch site.
Preparation of experiment at launch site, including starting of cultures, inoculation of agar plates / microcapsules, sealing of containers, then handover of experiment at ~L-14h;
Ambient upload of experiment and transfer to ISS (ambient temperature).
Photography of growth of fungal cultures on agar at intervals in-flight (two intervals preferred, one minimum).
Download of short duration samples at ambient temperature (~10-11 days total flight duration).

Expected results:
- Different orientation of hyphae, conidiophore and possible spores in microgravity
- Establishing compatibility/incompatibility of fungal life with cosmic radiation; Possibility to use cosmic radiations for decontamination of organic and/or inorganic substrates in space conditions and on the Earth.
- Different data-bases on different sensitivity and survival of spores to cosmic radiation according to species biology
- Phenotypic and biochemical mutations
- Changes in spores walls which can be identified using AFM
- Changes in colonization of specific substrate
- Finding specific protectors and strategies to survive for different fungi species to cosmic radiation
- Establishing a possible proliferation of potential forward contaminants transferred from Earth on spacecraft
- To develop a model of survival of spore forming fungi during transport to the other planetary bodies
- Design of a user friendly microcapsules and container for radiological research in space
- Passive ionizing radiation dosimeters, exposed to the space radiation environment close to the experiment samples

Growth of Ulocladium chartarum colonies takes place in real microgravity conditions with different rates that are correlated with the age of the colonies. Different growth rates were observed for the aerial and for the submerged mycelium. Growth of aerial mycelium has a high rate in flight and ground controls up to Flight Day 5, then becomes lower and stops between Flight Day 8-9. Sporulation takes place in flight and ground but it is less abundant compared to ground and laboratory control.

Integration of the microcapsules in the biocontainers shows a negative effect on the growth and on the sporulation in comparison with the laboratory control; new ground experiments will be made to acquire more information.

Microgravity reduces the rate of growth of aerial mycelium and stimulates the growth of submerged mycelium. The CFS-A experiment demonstrates that fungi as biodeteriogens and biodegraders are able to grow in microgravity, such as inside the ISS where substrates are humid. In gravity the growth of fungi can be identified from the size of the macroscopical colonies. However, in microgravity the development of exclusively submerged mycelium, after aerial growth is stopped, needs microscopic methods to be identified. Submerged mycelium is able to synthesise extra cellular enzymes which decompose the substrate where it is grown. So different astronaut food and materials (mostly those composed of organics) can be decomposed in humid conditions. Spreading of fungus in microgravity is very low because submerged mycelium is not able to make spores. This process can take place in the environment, following the growth pattern of the hyphae and branch tips (it is a way of colonization of the substrate), in cases such as when it comes to direct contact to the astronauts, by transporting contaminated item to other items which are not contaminated. Real colonies can be developed on ISS only if the submerged mycelium sense mechanical pressure. In such cases colonies can be visible by the astronauts, and therefore they will require urgent decontamination.

For the dry spore samples the spores chosen for the CFS-A experiment were still viable after 5 months in microgravity. Ulocladium chartarum spores are more resistant from a viability point of view then Basipetospora halophila and Cladosporium herbarum spores but less then Aspergillus niger spores. Aspergillus niger spores were more than 91% viable on all types of wafers. Ulocladium chartarum CM-1 spores have a good viability (87-92%) after 5 months as dried samples in weightlessness. Basipetospora halophila spores have a lower viability on ISS then on ground, which could suggest that white spores are more sensitive to ISS environment than black spores. Basipetospora halophila spores also showed a lower viability on the silica wafers then on plastic wafers and no viability on iron wafers probably due to a strong oxidation of iron wafer in contact with salts removed from the nutrient. If by chance food preserved in salt that is contaminated with this species, is brought to space, a degradation process could start. Salted food is good nutrient for fungal spores in space. Viability of Cladosporium herbarum spores is lower on the silica wafers then on plastic wafers. Again iron wafers did not sustain viability of the spores. The main reason is most probably the toxic effect of iron ions. High viability of Aspergillus niger and Ulocladium chartarum spores on iron wafers showed that these species could grow on iron surfaces covered with small quantities of carbohydrates on ISS. These results will feed into the wafers chosen for a follow-up experiments.
[1]  
I. Gomoiu, E. Chatzitheodoridis, D. Hasegan, (2011), "30‐Days Accomplishment Report: Growth and Survival of Coloured Fungi in Space", IBB‐CFSA‐PR 003.
[2]  
I. Gomoiu, E. Chatzitheodoridis, D. Hasegan, (2011), "30‐Days Accomplishment Report for 5 months duration of spores on ISS", IBB‐CFSA‐PR 004.
[3]  
(2013), "The Effect of Spaceflight on Growth of Ulocladium chartarum Colonies on the International Space Station", PLoS ONE, 8, 4, doi:10.1371/journal.pone.0062130, pp. e62130.
click on items to display

Colored fungi will be flown and allowed to develop in microgravity conditions. The effects of microgravity on development, metabolism and sporulation of the cultivated fungi will be measured. The BASE-A / YING-A (in picture) illustrates the experiment concept. Two sets of samples will be flown. One set containing live cultures and dried spores will aim at monitoring the growth, development and metabolism of fungi over a short duration mission in space. The second set will consist of dried spores only and will be stowed in space for at least 5 months to assess the combined impact of microgravity and radiation on sporulation.

Detailed Experiment Timeline and associated Functional Objectives.
 
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