Maximum carbon isotope fractionation in photosynthesis by blue-green algae and a green alga
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Algae growth in a micro-gravity environment is an important factor in supporting man's permanent presence in space. Algae can be used to produce food, oxygen, and pure water in a manned space station. A space station is one example of a situation where a Controlled Ecological Life Support System (CELSS) is imperative. In setting up a CELSS with an engineering approach at the Aerospace department of the University of Colorado, questions concerning algae growth in micro-g have arisen. The Get Away Special (GAS) Fluids Management project is a means through which many questions about the effects of a micro-g environment on the adequacy of growth rates, the viability of micro-organisms, and separation of gases and solids for harvesting purposes can be answered. In order to be compatible with the GAS tests, the algae must satisfy the following criteria: (1) rapid growth rates, (2) sustain viability over long periods of non-growth storage, and (3) very brief latency from storage to rapid growth. Testing indicates that the overall growth characteristics of Anacystis Nidulans satisfy the specifications of GAS's design constraints. In addition, data acquisition and the method of growth instigation are two specific problems being examined, as they will be encountered in interfacing with the GAS project. Flight testing will be two-fold, measurement of algae growth in micro-g and separation of algae from growth medium in an artificial gravitation field. Post flight results will provide information on algae viability in a micro-g environment as reflected by algal growth rates in space. Other post flight results will provide a basis for evaluating techniques for harvesting algae. The results from the GAS project will greatly assist the continuing effort of developing the CELSS and its applications for space.
Algae has abundant potential spaceflight applications to enable future crewed missions and habitation beyond low-Earth orbit, including production of essential nutrients, oxygen, and biofuels. Implementing algae production in space requires understanding how different algae species respond to spaceflight stressors such as microgravity and radiation over a realistic production time course. Space Algae-2 is focusing on Arthrospira platensis, a filamentous cyanobacteria commonly known as Spirulina. A. platensis is a human nutritional supplement on Earth that is a good source of essential amino acids, β-carotene, thiamin, riboflavin, and antioxidants. The Space Algae-2 experiment plans to grow and passage A. platensis continuously aboard the International Space Station for six months. The algae produced will be analyzed with multi-omics profiling to monitor for genetic and phenotype stability in the spaceflight environment. This presentation will report the results from pre-flight science verification tests of the concept of operations for Space Algae-2. The purpose of these tests was to determine if the proposed crew operations for serial passages, sample collection, and sample preservation are feasible to meet science requirements for axenic culturing and preservation of DNA, RNA, protein, and nutritional metabolites.
Method and system for estimating a growth stage of an alga in an ambient fluid. Measured light beam absorption or reflection values through or from the alga and through an ambient fluid, in each of two or more wavelength sub-ranges, are compared with reference light beam absorption values for corresponding wavelength sub-ranges for in each alga growth stage to determine (1) which alga growth stage, if any, is more likely and (2) whether estimated lipid content of the alga is increasing or has peaked. Alga growth is preferably terminated when lipid content has approximately reached a maximum value.
An intertidal Chlorella-like alga Mychonastes desiccatus Brown sp. nova, capable of forming achlorophyllous desiccation-resistant cysts, has been grown in unialgal culture. This small alga was first isolated from a dried sample of a well-studied microbial mat. The mat, located at North Pond, Laguna Figueroa, San Quintin, Baja California, Mexico, is a vertically-stratified microbial community which forms laminated sediments. Morphology, pigment composition and G+C content are within the range typical for the genus Chlorella s. 1. Unlike other chlorellae, however, upon desiccation M. desiccatus forms an achlorophyllous, lipid-filled cyst (thick-walled resting stage) in which no plastid is evident. Rewetting leads to chloroplast differentiation, excystment and recovery of the fully green alga. During desiccation, sporopollenin is deposited within a thickening cell wall. Encystment cannot be induced by growth in the dark. The formation of desiccation-induced cysts allows the alga to survive frequent and intermittent periods of dryness. These chlorellae tolerate wide ranges of acidity and temperature; they both grow and form cysts in media in which sodium ions are replaced with potassium. Although the cysts tolerate crystalline salts, the cell grow optimally in concentrations corresponding from three-quarters to full-strength seawater.
The effect of microgravity on living organisms during space flight has been a topic of interest for some time, and a substantial body of knowledge on the subject has accumulated. Despite this, comparatively little information is available regarding the influence of microgravity on algae, even though it has been suggested for long duration flight or occupancy in space that plant growth systems, including both higher plants and algae, are likely to be necessary for bioregenerative life support systems. High-Aspect-Ratio Rotating-Wall Vessel or HARV bioreactors developed at Johnson Space Center provide a laboratory-based approach to investigating the effects of microgravity on cellular reactions. In this study, the HARV bioreactor was used to examine the influence of simulated microgravity on the growth and metabolism of the green alga, Chlorella pyrenoidosa. After the first 2 days of culture, cell numbers increased more slowly in simulated microgravity than in the HARV gravity control; after 7 days, growth in simulated microgravity was just over half (58%) that of the gravity control and at 14 days it was less than half (42%). Chlorophyll and protein were also followed as indices of cell competence and function; as with growth, after 2-3 days, protein and chlorophyll levels were reduced in modeled microgravity compared to gravity controls. Photosynthesis is a sensitive biochemical index of the fitness of photosynthetic organisms; thus, CO2-dependent O2 evolution was tested as a measure of photosynthetic capacity of cells grown in simulated microgravity. When data were expressed with respect to cell number, modeled microgravity appeared to have little effect on CO2 fixation. Thus, even though the overall growth rate was lower for cells cultured in microgravity, the photosynthetic capacity of the cells appears to be unaffected. Cells grown in simulated microgravity formed loose clumps or aggregates within about 2 days of culture, with aggregation increasing over time. Presently, the basis for, or significance of, the cell aggregation is unknown. The results from this study suggest that cell growth and morphological characteristics of green algae may be altered by culture in simulated microgravity. The data obtained to date should provide a solid basis for additional experimentation regarding the influence of modeled microgravity on cell morphology, physiological activity, protein production and possibly gene expression in algal and plant cell systems. The final aim of the study is to provide useful information to elucidate the underlying mechanism for the biological effects of microgravity on cells.
Methods for producing hydrocarbons, including oil, by processing algae and/or other micro-organisms in an aquatic environment. Flexible bags (e.g., plastic) with CO.sub.2/O.sub.2 exchange membranes, suspended at a controllable depth in a first liquid (e.g., seawater), receive a second liquid (e.g., liquid effluent from a "dead zone") containing seeds for algae growth. The algae are cultivated and harvested in the bags, after most of the second liquid is removed by forward osmosis through liquid exchange membranes. The algae are removed and processed, and the bags are cleaned and reused.
Plants and microbes can be used for biological support of crewed space missions. The radiation and microgravity environment of spaceflight is expected to increase genetic mutation of all organisms. It is essential to understand how spaceflight impacts mutation rates in photosynthetic organisms to enable appropriate countermeasures and ensure productivity during long duration and deep space missions. The Space Algae flight experiments to the International Space Station (ISS) are studying the genomic stability of microalgae that could potentially be used in biological life support systems. Space Algae-1 grew ultraviolet light mutagenized Chlamydomonas reinhardtii in the VEGGIE plant growth chamber for approximately 40 mitotic generations over one month on the ISS. Whole genome sequencing from pooled cell samples every 10 generations revealed that spaceflight cultures had an ~50% increase in DNA polymorphisms relative to ground controls. These mutations had a novel base substitution signature and suggested a risk that microalgae may be unstable for long-term production in space. Space Algae-2 is focusing on the edible cyanobacterium Arthrospira platensis, commonly known as Spirulina. This experiment seeks to grow serial cultures to allow the organism to evolve in long-term spaceflight. Biological responses of the cells to spaceflight will be assessed with multi-omics analyses to determine mutation load, gene/protein expression, metabolic/nutritional composition, and cell morphology.
Plants and microbes can be used for biological support of crewed space missions. The radiation and microgravity environment of spaceflight is expected to increase genetic mutation of all organisms. It is essential to understand how spaceflight impacts mutation rates in photosynthetic organisms to enable appropriate countermeasures and ensure productivity during long duration and deep space missions. The Space Algae flight experiments to the International Space Station (ISS) are studying the genomic stability of microalgae that could potentially be used in biological life support systems. Space Algae-1 grew ultraviolet light mutagenized Chlamydomonas reinhardtii in the VEGGIE plant growth chamber for approximately 40 mitotic generations over one month on the ISS. Whole genome sequencing from pooled cell samples every 10 generations revealed that spaceflight cultures had an ~50% increase in DNA polymorphisms relative to ground controls. These mutations had a novel base substitution signature and suggested a risk that microalgae may be unstable for long-term production in space. Space Algae-2 is focusing on the edible cyanobacterium Arthrospira platensis, commonly known as Spirulina. This experiment seeks to grow serial cultures to allow the organism to evolve in long-term spaceflight. Biological responses of the cells to spaceflight will be assessed with multi-omics analyses to determine mutation load, gene/protein expression, metabolic/nutritional composition, and cell morphology.
There are numerous applications for microalgae in spaceflight missions and on Earth, such as oxygen production, carbon dioxide removal, nutrition, wastewater processing, and biofuel production. Space Algae-2 aims to test the genetic stability of Arthrospira platensis, commonly known as spirulina, during six-months of continuous culture on the International Space Station. Long-duration exposure to ionizing radiation and microgravity may impact growth, nutrient composition, and genetic stability. The high protein, vitamin, antioxidant content, and radiation resistance make spirulina a promising candidate for bioregenerative life support systems. A concept of operations was developed to grow and harvest algal biomass in space. Preflight testing experiments optimized conditions for an extended growth period in a gas permeable bioreactor bag. Preflight and post-harvest storage methods were developed in addition to a novel cryopreservation method. After sample return, multi-omics analysis will be conducted to determine the mutation rate, gene expression, and the protein and metabolite profile. The concept of operations for Space Algae-2 was tested during a lunar mission simulation within a semi-controlled environment. During a six-day lunar analog mission at the Hawai’i Space Exploration Analog and Simulation (HI-SEAS) A. platensis was successfully grown using flight-like hardware. The cyanobacteria were harvested and used to supplement bread as an example of spirulina biomass utilization. Overall, the data collected from Space Algae-2 will inform potential bioengineering of spirulina for space and terrestrial applications.
There are numerous applications for microalgae in spaceflight missions and on Earth, such as, oxygen production, carbon dioxide removal, nutrition, wastewater processing, and biofuel production. Space Algae-2 aims to test the genetic stability of Arthrospira platensis, commonly known as spirulina, during six-months of continuous culture in spaceflight on the International Space Station. Long-duration exposure to ionizing radiation and microgravity may impact growth, nutrient composition, and genetic stability. The high protein, vitamin, antioxidant content, and radiation resistance make spirulina a promising candidate for bioregenerative life support systems during long-duration missions. A concept of operations was developed to grow and harvest algal biomass in space. Preflight testing experiments were conducted to optimize conditions for an extended growth period in a gas permeable bioreactor bag. Preflight and post-harvest storage methods were developed in addition to a novel cryopreservation method. After sample return, multi-omics analysis will be conducted to determine the mutation rate, gene expression, and protein and metabolite profile. The concept of operations for Space Algae-2 was tested at HI-SEAS (Hawai’i Space Exploration Analog and Simulation) during a six-day lunar analog mission (EMMIHS23, EuroMoonMars, International MoonBase Alliance, HI-SEAS, 2023). A. platensis was grown in the semi-controlled environment using flight-like hardware and solar powered LED lights. Then, the biomass was harvested and used to supplement bread as an example of A. platensis utilization. Overall, the data collected from Space Algae-2 will inform potential bioengineering of spirulina for space and terrestrial applications.
There are numerous applications for microalgae in spaceflight missions and on Earth, such as oxygen production, carbon dioxide removal, nutrition, wastewater processing, and biofuel production. Space Algae-2 aims to test the genetic stability of Arthrospira platensis, commonly known as spirulina, during six-months of continuous culture on the International Space Station. Long-duration exposure to ionizing radiation and microgravity may impact growth, nutrient composition, and genetic stability. The high protein, vitamin, antioxidant content, and radiation resistance make spirulina a promising candidate for bioregenerative life support systems. A concept of operations was developed to grow and harvest algal biomass in space. Preflight testing experiments optimized conditions for an extended growth period in a gas permeable bioreactor bag. Preflight and post-harvest storage methods were developed in addition to a novel cryopreservation method. After sample return, multi-omics analysis will be conducted to determine the mutation rate, gene expression, and the protein and metabolite profile. The concept of operations for Space Algae-2 was tested during a lunar mission simulation within a semi-controlled environment. During a six-day lunar analog mission at the Hawai’i Space Exploration Analog and Simulation (HI-SEAS) A. platensis was successfully grown using flight-like hardware. The cyanobacteria were harvested and used to supplement bread as an example of spirulina biomass utilization. Overall, the data collected from Space Algae-2 will inform potential bioengineering of spirulina for space and terrestrial applications.
A technique for remote measurement of chlorophyll a density and determination of algae color group is described. In vivo fluorescence measurements of chlorophyll a for four color groups of algae (green, golden-brown, red, and blue-green) are described, and representative spectra are shown. The lidar equation is developed for the general case of a mixture of color groups showing the need for multicolor excitation. The lidar instrument which has been designed and fabricated at Langley Research Center for helicopter flights over surrounding portions of the Chesapeake Bay is described. The lidar package contains a unique four-color dye laser which permits multicolor excitation of chlorophyll a fluorescence from the various color groups of algae.
A theoretical and experimental analysis of laser induced fluorescence for remote detection of chlorophyll A in living algae and phytoplankton is presented. The fluorescent properties of various species of algae representative of the different color groups are described. Laboratory measurements of fluorescent scattering cross sections is discussed and quantitive data presented. A scattering matrix model is developed to demonstrate the essential requirement of multiwavelength laser excitation in order to make accurate quantitative measurements of chlorophyll A concentration when more than one color group of algae is present in the water. A practical airborne laser fluorosensor design is considered and analysis of field data discussed. Successful operation of the Langley ALOPE (airborne LIDAR oceanographic probing experiment) system is described and field measurements presented. Accurate knowledge of alpha, the optical attenuation coefficient of the water, is shown to be essential for quantitative analysis of chlorophyll A concentration. The feasibility of remotely measuring alpha by laser radar is discussed.
The development and performance of single- and multiple-wavelength laser fluorosensor systems for use in the remote detection and quantification of algae are discussed. The appropriate equation for the fluorescence power received by a laser fluorosensor system is derived in detail. Experimental development of a single wavelength system and a four wavelength system, which selectively excites the algae contained in the four primary algal color groups, is reviewed, and test results are presented. A comprehensive error analysis is reported which evaluates the uncertainty in the remote determination of the chlorophyll a concentration contained in algae by single- and multiple-wavelength laser fluorosensor systems. Results of the error analysis indicate that the remote quantification of chlorophyll a by a laser fluorosensor system requires optimum excitation wavelength(s), remote measurement of marine attenuation coefficients, and supplemental instrumentation to reduce uncertainties in the algal fluorescence cross sections.
Solar cells (2 cm by 2 cm (AlGa) As-GaAs cells) were fabricated and then subjected to irradiation at normal incidence by electrons. The influence of junction depth and n-type buffer layer doping level on the cell's resistance to radiation damage was investigated. The study shows that (1) a 0.3 micrometer deep junction results in lower damage to the cells than does a 0.5 micrometer junction, and (2) lowering the n buffer layer doping density does not improve the radiation resistance of the cell. Rather, lowering the doping density decreases the solar cell's open circuit voltage. Some preliminary thermal annealing experiments in vacuum were performed on the (AlGa)As-GaAs solar cells damaged by 1-MeV electron irradiation. The results show that cell performance can be expected to partially recover at 200 C with more rapid and complete recovery occurring at higher temperature. For a 0.5hr anneal at 400 C, 90% of the initial power is recovered. The characteristics of the (AlGa)As-GaAs cells both before and after irradiation are described.
Data are given of a comparative study of the growth and development as well as the characteristics of the biomass of the C. Lipolytica yeast according to the content of raw protein, protein, lipids, vitamins in the B group, and residual hydrocarbons during growth in media with de-aromatized gas-condensate FNZ as the carbon source with aqueous and alcohol extracts of S. acuminatus as the biostimulants. It is shown that the decoction and aqueous extract of green algae has the most intensive stimulating effect on the yeast growth. When a decoction of algae is added to the medium, the content of residual hydrocarbons in the biomass of C. lipolytica yeast is reduced by 4%; the quantity of protein, lipids, thamine and inositol with replacement of the yeast autolysate by the decoction of algae is altered little.
One of the intriguing facets of the Cretaceous-Tertiary extinction is the apparently selective pattern of mortality amongst taxa. Some groups of organisms were severely affected and some remained relatively unscathed as they went through the K/T boundary. While there is argument concerning the exact interpretation of the fossil record, one of the best documented extinctions at the Cretaceous-Tertiary boundary is that of the calcareous nannoplankton. These organisms include coccolithic algae and foraminiferans. Attempts to explain their decline at the K/T boundary center around chemistry which could affect their calcium carbonate shells while leaving their silica-shelled cousins less affected or unaffected. Two environmental consequences of an extraterrestrial body impact which were suggested are the production of large quantities of nitrogen oxides generated by the shock heating of the atmosphere and the possible rise in CO2 from the dissolution of CaCO3 shells. Both of these phenomena would acidify the upper layers of the oceans and bodies of freshwater not otherwise buffered. The effects of nitric acid, carbon dioxide, or both factors on the growth and reproduction of calcareous marine coccoliths and non-calcareous marine and freshwater species of algae were considered. These experiments demonstrate that nitric acid and carbon dioxide have significant effects on important aspects of the physiology and reproduction of modern algae representative of extinct taxa thought to have suffered significant declines at the Cretaceous-Tertiary boundary. Furthermore, calcareous species showed more marked effects than siliceous species and marine species tested were more sensitive than freshwater species.