ERA-experiment "Space Biochemistry".

The general goal of the experiment was to study the response of anhydrobiotic (metabolically dormant) microorganisms (spores of Bacillus subtilis, cells of Deinococcus radiodurans, conidia of Aspergillus species) and cellular constituents (plasmid DNA, proteins, purple membranes, amino acids, urea) to the extremely dehydrating conditions of open space, in some cases in combination with irradiation by solar UV-light. Methods of investigation included viability tests, analysis of DNA damages (strand breaks, DNA-protein cross-links) and analysis of chemical effects by spectroscopic, electrophoretic and chromatographic methods. The decrease in viability of the microorganisms was as expected from simulation experiments in the laboratory. Accordingly, it could be correlated with the increase in DNA damages. The purple membranes, amino acids and urea were not measurably effected by the dehydrating condition of open space (in the dark). Plasmid DNA, however, suffered a significant amount of strand breaks under these conditions. The response of these biomolecules to high fluences of short wavelength solar UV-light is very complex. Only a brief survey can be given in this paper. The data on the relatively good survival of some of the microorganisms call for strict observance of COSPAR Planetary Protection Regulations during interplanetary space missions.     

Biological degradation and composition of sweet potato biomass - errata

Many challenges are presented by biological degradation in a bioregenerative Controlled Ecological Life Support System as envisioned by the U.S. National Aeronautics and Space Administration. In studies conducted with biodegradative microorganisms indigenous to sweetpotato fields, it was determined that a particle size of 75 microns and incubation temperature of 30°C were optimal for degradation. The composition of the biomass and characterization of plant nutrient solution indicated the presence of potential energy sources to drive microbial transformations of plant waste. Selected indigenous soil isolates with ligno-cellulolytic or sulfate-reducing ability were utilized in biological studies and demonstrated diversity in their ability to reduce sulfate in solution and to utilize alternative carbon sources: a lignin analog 4-hydroxy, 3-methoxy cinnamic acid, cellulose, arabinose, glucose, sucrose, mannitol, galactose, ascorbic acid.     

Engineering strategies for the design of plant nutrient delivery systems for use in space: approaches to countering microbiological contamination.

Microbiological contamination of crops within space-based plant growth research chambers has been postulated as a potentially significant problem. Microbial infestations; fouling of Nutrient Delivery System (NDS) fluid loops; and the formation of biofilms have been suggested as the most obvious and important manifestations of the problem. Strict sanitation and quarantine procedures will reduce, but not eliminate, microbial species introduced into plant growth systems in space habitats. Microorganisms transported into space most likely will occur as surface contaminants on spacecraft components, equipment, the crew, and plant-propagative materials. Illustrations of the potential magnitude of the microbiological contamination issue will be drawn from the literature and from documentation of laboratory and commercial field experience. Engineering strategies for limiting contamination and for the development of countermeasures will be described. Microbiological control technologies and NDS hardware will be discussed. Configurations appropriate for microgravity research facilities, as well as anticipated bio-regenerative life support system implementations, will be explored. An efficiently designed NDS, capable of adequately meeting the environmental needs of crop plants in space, is considered to be critical in both the research and operational domains. Recommended experiments, tests, and technology developments, structured to allow the development of prudent engineering solutions also will be presented.     

Plant growth and gas balance in a plant and mushroom cultivation system.

In order to obtain basic data for construction of a plant cultivation system incorporating a mushroom cultivation subsystem in the CELSS, plant growth and atmospheric CO2 balance in the system were investigated. The plant growth was promoted by a high level of CO2 which resulted from the respiration of the mushroom mycelium in the system. The atmospheric CO2 concentration inside the system changed significantly due to the slight change in the net photosynthetic rate of plants and/or the respiration rate of the mushroom when the plant cultivation system combined directly with the mushroom cultivation subsystem.     

Molecular analysis of mutagenesis by high LET radiation.

Mutation induction by high linear energy transfer [LET] alpha particles and gamma-rays was scored in the human hamster hybrid [AL] cells. Southern blotting technique was used to analyse the molecular changes in the DNA from both the HGPRT- and S1- mutants. Dose dependent mutagenesis in the AL cells irradiated with the charged particles was higher by almost 20 fold at the S1 than the corresponding HGPRT locus. Southern analysis of the mutants induced by the high LET particles showed mostly multilocus deletion at both the HGPRT and S1 genes.     

Gravitational response of the slime mold Physarum.

The acellular slime mold Physarum polycephalum is used as a model system to investigate the graviresponse of single cells which possess no receptors specialized for the perception of gravity. To obtain insights into the gravity-signal transduction mechanism the light response of the cell is used: Macroplasmodia of the slime mold show clear geo- and phototaxes. Gravity increases and white light decreases transiently the contraction frequency of plasmodial strands whereby both responses follow the same time pattern. Since mitochondria play a major role in changing the contraction rhythm in response to light and gravity stimuli, the simultaneous and subsequent inductions of the opposing light and gravity responses and their mutual influences on one another were investigated. The experiments were performed in weightlessness (0 g)--simulated on the fast-rotating clinostat as well as in actual weightlessness during the IML-1 Space Shuttle mission. The results indicate that mitochondria (chondriome) are part of the acceleration-stimulus reaction chain in Physarum. Two models for a direct gravireceptor mechanism are discussed.     

Calculations and observations of solar particle enhancements to the radiation environment at aircraft altitudes.

Solar particle events can give greatly enhanced radiation at aircraft altitudes, but are both difficult to predict and to calculate retrospectively. This enhanced radiation can give significant dose to aircrew and greatly increase the rate of single event effects in avionics. Validation of calculations is required but only very few events have been measured in flight. The CREAM detector on Concorde detected the event of 29 September 1989 and also four periods of enhancement during the events of 19-24 October 1989. Instantaneous rates were enhanced by up to a factor ten compared with quiet-time cosmic rays, while flight-averages were enhanced by up to a factor six. Calculations are described for increases in radiation at aircraft altitudes using solar particle spectra in conjunction with Monte Carlo radiation transport codes. In order to obtain solar particle spectra with sufficient accuracy over the required energy range it is necessary to combine space data with measurements from a wide range of geomagnetically dispersed, ground-level neutron monitors. Such spectra have been obtained for 29 September 1989 and 24 October 1989 and these are used to calculate enhancements that are compared with the data from CREAM on Concorde. The effect of cut-off rigidity suppression by geomagnetic activity is shown to be significant. For the largest event on record on 23 February 1956, there are no space data but there are data from a number of ground-level cosmic-ray detectors. Predictions for all events show very steep dependencies on both latitude and altitude. At high latitude and altitude (17 km) calculated increases with respect to cosmic rays are a factor 70 and 500 respectively for 29 September 1989 and 23 February 1956. The levels of radiation for high latitude, subsonic routes are calculated, using London to Los Angeles as an example, and can exceed 1 mSv, which is significantly higher than for Concorde routes from Europe to New York. The sensitivity of the calculations to spectral fitting, geomagnetic activity and other assumptions demonstrates the requirement for widespread carriage of radiation monitors on aircraft.     

Survival and alteration of the plasmid-containing microorganism Escherichia coli Z905/pPHL7 introduced into manmade closed aquatic microcosms.

It has been demonstrated that the transgenic microorganism Escherichia coli Z905/pPHL7 (AprLux+) can exist for a long time at an elevated concentration of mineral salts. The microorganism was introduced into microcosms with sterile brackish water (salinity variable from 21 to 22 g l-1) taken from Lake Shira (Khakasia, Russia). The survival of the microorganism was estimated both by measuring the growth of the colonies on solid nutrient media and by the bioluminescence exhibited by the transgenic strain in samples from the microcosms and in the enrichment culture with the added selective factor-ampicillin (50 micrograms/ml). In the enrichment culture, the bioluminescent signal was registered through the 160-day experiment. It has been shown that in the closed microcosms with brackish water the E. coli strain becomes heterogeneous in its ampicillin resistance. The populations of the transgenic strain were mainly represented by isolates able to persist in the medium containing 50 micrograms/ml, but there were also the cells (about 10%) with the threshold of ampicillin resistance not more than 0.05 micrograms/ml. Thus, it was shown that in the microcosms with brackish water and in the absence of the selective factor the transgenic strain survives and retails the recombinant plasmid.     

Microgravity particle research on the space station: the Gas-Grain Simulation Facility.

In the gravitational field on Earth, the large settling rate of micron-sized particles and the effects of gravity-induced convection prohibit many interesting studies of phenomena such as coagulation, collisions, and mutual interactions of droplets, dust grains and other particles. Examples of exobiology experiments involving these phenomena are the simulation of organic aerosol formation in Titan's atmosphere, studies of the role of comets in prebiotic chemical evolution, and simulations of carbon grain interactions in various astrophysical environments. The Gas-Grain Simulation Facility (GGSF) is a proposed Earth-orbital laboratory that will allow present ground-based experimental programs which study processes involving small particles and weak interactions to be extended to a new domain. Physics issues that scientists wishing to propose GGSF experiments must consider are reviewed in this paper. Specifically, coagulation, motion in gases and vacua, and wall deposition of particles in a microgravity environment are discussed.     

Productivity and food value of Amaranthus cruentus under non-lethal salt stress.

Stress effects from the accumulation of metal salts may pose a problem for plants in closed biological systems such as spacecraft. This work examined the effects of salinity on growth, photosynthesis and carbon allocation in the crop plant, Amaranthus. Plants were germinated and grown in modified Hoagland's solution with NaCl concentrations of 0 to 1.0%. Plants received salt treatments at various times in development to assess effects on particular life history phases. For Amaranthus cruentus, germination, vegetative growth, flowering, seed development and yield were normal at salinities from 0 to 0.2%. Inhibition of these phases increased from 0.2 to 0.4% salinity and was total above 0.5%. 1.0% salinity was lethal to all developmental phases. Onset of growth phases were not affected by salinity. Plants could not be adapted by gradually increasing salinity over days or weeks. Water uptake increased, while photosynthetic CO2 uptake decreased with increasing salinity on a dry weight basis during vegetative growth. Respiration was not affected by salinity. After flowering, respiration and photosynthesis decreased markedly, such that 1.0% NaCl inhibited photosynthesis completely. Protein levels were unchanged with increasing salinity. Leaf starch levels were lower at salinities of 0.5% and above, while stem starch levels were not affected by these salinities. The evidence supports salt inhibition arising from changes in primary biochemical processes rather than from effects on water relations. While not addressing the toxic effects of specific ions, it suggests that moderate salinity per se need not be a problem in space systems.