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NASA CONTRACTOR REPORT 166615 (YASA-CE-166615) PBOELLBS ASSOCIATED VITU li85-164E9 TBE UTILIZATION OF ALGAE 16 EICCEGENEBAIIVE LIFE SUPPORT SYSTUS (New Elampshire Univ.) 24 p HC AO2/HF A01 CSCL 06C Unc la E G3/54 13240 -ablems Associated with the Utilization of algae in Bioregenerative Life Support Systems Maurice Averner Yarcus Karel Richard Radmer NASA Cooperative Agreement NCC 2-210 November 1984 NASA CONTRACTOR REPORT 166615 Problems Associated with the Utilization of Algae in Bioregonerative Life Support Systems Maurice Averner Complex Systems Research Center University of New Hampshire Durham, NH 03824 Marcus Karel Dept. of Nutrition and Food Science Xassachusetts Institute of Technology Cambridge, MA 02139 Richard Radmer Martin Marietta Laboratories 1450 S. Rolling Road Baltimore, MD 21227 Prepared for Ames Research Center Under NASA Cooperative Agreement NCC 2-210 MSA National Aeronautics and Space Adminlstration Ames Research Center Moffett Field, California94035 INTRODUCTION The systems presently used for human life support in space require that food, water, and oxygen be stored, that excess atmospheric carbon dioxide be removed, and that other human wastes be collected and stored. Lengthy missions or large crews dictate that large masses of consumable supplies be taken along or resupplied. L'lti- mately, a point will be reached where the regeneration of consumables will be econorn- ically competitive with the cost of their initial transport or resupply. It is possible :o envision a number of systems that in theory nt least would be capable Df recycling human waste gases, liquids, and solids, and regenerating food, oxygen, and potable water. Among such theoretical systems, those based tin the cse of biological processes are very attractive and have been the subject of extensive study in the Controlled Environment Life Support System (CELSS) program. This SASA- sponsored program is directed at studying the feasibility of constructing a bioregen- erative life-support system for use in extraterrestrial environments. X bioregenera- tive life-support system can be described as one that uses rigorously controlli.3 acd integrated biological and physicochemical processes to regeneraie the supplies con- sumed in life-support systems. A biological process that is likely to be central to the successful deveIop~.;.nt of such a system is photosynthesis. Photosynthesis has the potential for silnultarie- ously carrying out most of the major functions of a life-support system: production of food, of oxygen, arid of potable water, and the removal of carbon dioxide. .bong those organisms capable of photosynthesis, there are two kinds of microorganisms - the green algae (Chlorophyta), and the blue-green algae (Cyanophyta) -that have a number of characteristics that make them attractive candidates for inclusion in a bioregenerative system. These algae grow rapidly, their metabolism can be controlled, they produce a high ratio of edible to nonedible biomass, and their gas-exchange characteristics are compatible with human requirements. In addition, many strai?is of blue-green algae can convert gaseous nitrogen into ammonium, a biologically usehi iorm of nitrogen. Unfortmately, there are a number of problems associated with the use of micro- algae in life-support syster;.s. For example , there are questions about the adequac!- and acceptability of algal-derived food, about harvesting and prozessing of the algAL, md about the long-term stability cf the algal cultures. And although ground-bas2d experinentation will be essential for identifying and resolving some of these pros- lems, others can only be approached by appropriate spaceflight experimentation. With this in mind, a workshop was held at the Ames Research Center on Sovember 28-29, 1983, for the purpose of bringing together a group of experts to discuss two major issues: (1) the identification of the major problems associated with the use of microalgae In a bioregenerative life-support system; and (2) the identification of algae-related research issues that must be addressed through spaceflight experimentation. The contributors to this Workshop wer: scientists with expertise in a variet>- of pertinent areas, including algal growth and physiology, the production of food frcni nonconventional food sources, and human nutrition. 1 WORKSHOP AGENDA Problems Associated With the Utilization of Algae in Regenerative Life-support Systems for Space Habitation NASA Ames Research Center Moffett Field, California 94035 Room 361, Building 239 (Life Sciences) Monday, November 28, 1983 Session I (9:OO to 11:30) Presentation of the Problem Chair: Dr. M. M. Averner 9:OO - 9:05 Dr. H. ?. Klein Opening Remarks 9:OS - 9:20 Dr. R. D. MacElroy T??e Status and Role of Regenerative Life-support Research in NASA 9:20 - 9:50 Dr. M. M. Averner The Potential Functions of Algae in Regenerative Life-support Systems 9~50- 10~30 Dr. Richard Radmer The State of the Art in Sustained Culture of Algae under Space-Related Conditions 10:30 - 10~45 Coffee Break 10~45- 11:30 Dr. Marcus Karel The Problem of the Conversion of Nonconventional Substrates to Hunan Food 11:30 - 1:OO LUNCH Session I1 (1:OO to 3:45) Round-Table Discussion of Problems of Algal Utilization Moderators: Drs. Radmer and Karel 3:45 - 4:OO Coffee -Session 111 (4:OO to 5:OO) Space-Flight Experiments Utilizing Algae Chair: Dr. Lester Packer 2 4:OO - 4:20 Introduction Dr. M. M. Averner 4:20 - 4:50 Opportunities for and Constraints on Space Flight Experimentation Dr. Edward Herek Tuesday, November 29, 1983 Session IV (9:OO to 12:OO) Identification of Research Issues to be Addressed by Space-Flight Experiments Moderators: Drs. MacElroy and Averner 12:OO - 1:30 Lunch Session V (1:30 to 3:30) Summary and Wrapup Chair: Dr. Marcus Karel 3 UTILIZATION OF ALGAE Algal Growth For the past 50 years or so, algal-culture studies have been closely inter- twined with studies of photosynthesis. Green algae, for example, Scenedesmus and Chlorella, have proved to be good models for higher plants. Several programs in applied algal culture have also been undertaken in the last 30 years, notably at the Carnegie Institute, and in programs sponsored by the United States Air Force and NASA on bioregenerative life support during the period of the mid-1950's to mid-1960's. Ob;riously, applied algal culture is not a new undertaking. Species of algae for space-related culture- Most of the work on algal physiology related to sustained culture under space-related conditions has been done on a rela- tively few species of green (eukaryotic) and blue-green (prokaryotic) algae. By far the most popular have been several species of Chlorella. which are consequently the best characterized organisms. The primary species currently considered for space- related applications are: (1) Chlorella (green) , several species; (2) Scenedesmus (green); (3) Anacystis (blue-green) = Synechococcus; and (4) Spirulina (blue-green). We should emphasize that there is no ideal alga. For example, Spirulina is not necessarily superior with respect to food value; it is simply the best studied in that respect. In some cases, there may be a definite trade-off between ease of cul- turing and harvesting; for example, Chlorella versus Spirulina. L. Packer and his colleagues suggest that the prokaryotic N2-fixing cyanobac- teria (blue-green) species deserve primary consideration, based on their following attributes: 1. For N2-fixing species, nutrient requirements are minimal, and no class of organisms exists in nature with simpler nutrient requirements. Except for inorganic nutrients, an aqueous environment, N2 or air, aerobic species will generally grow without auxillary substances. 2. Species can be selected that exhibit short generation times. 3. Species can be selected that exhibit a wide tolerance for environmental stresses, such as extreme drafts, high temperature, high salinity, and sudden expo- sure to such conditions. 4. These organisms utilize phycobiliproteins and phycobilisomes as light- harvesting systems, enabling these organisms to grow well in a broad range of light intensities between 600 and 650 nm, a range in which most other photosynthetic organisms will not grow well. 5. Strains are available that grow as unicellular organisms in suspension cul- ture, and filamentous suspension culture with or without sheath material. Strains can be selected that grow in aggregates, and in many configurations. It is possible to obtain isolates from nature that fulfill desired growth associations. 6. Many species produce gas vacuoles and will, therefore, be of interest for culturing in zero-gravity conditions. 4 7. Properties (5) and (6) can be exploited for purposes of harvesting or col- lecting cultures. 8. Under nitrogen limitation, the organisms will fix atmospheric Nz; thus, they can participate in a CELSS in which the nitrogen recycling and regeneration of biologically useful nitrogen and biomass is essential. 9. Most species produce a high content of proteins, in some cases as much as 70% of the total dry weight. 10. Under appropriate growth conditions, the relative proportion of carbohy- drates that can be accumulated in the form of glycogen,, lipid, or protein or any combination thereof can also be modulated, particularly by manipulating nitrogen limitations of growth. 11. If needed, cultures can be synchronized. Growth under steady-state condi- tions has been achieved with many species. Two other points made by R. Lewin: 1. Contamination could be reduced (if not eliminated) by growing algae capr!t?ic of tolerating extreme conditions, say high pH and high salinity (e.g., Spirulina)__ , low pH (e.g., Cyanidium and Spermatozopsis can tolerate pH less than 2 or 3), and high temperatures (e.g., Cyanidium and Mastigocladus can tolerate temperatures as high as 6OOC). 2. Algal products could be adjusted to have about 50% protein (in high-nitrsgcn media) or almost no organic nitrogen: glycerol (10%-30%from Dunaliella); mucilage (80% from Porphyridium grown on sponge in a flowing system); starch (perhaps 502 from green unicells); glucose, mannitol, or maltose from cells "permeabilized" (cf .