Evaluation of an anaerobic digestion system for processing CELSS crop residues for resource recovery.

Three bioreactors, connected in series, were used to process CELSS potato residues for recovery of resources. The first stage was an anaerobic digestor (8 L working volume; cow rumen contents inoculum; fed-batch; 8 day retention time; feed rate 25 gdw day-1) that converted 33% of feed (dry weight loss) to CO2 and "volatile fatty acids" (vfa, 83:8:8 mmolar ratio acetic:propionic:butyric). High nitrate-N in the potato residue feed was absent in the anaerobic effluent, with a high portion converted to NH4(+)-N and the remainder unaccounted and probably lost to denitrification and NH4+ volatilization. Liquid anaerobic effluent was fed to an aerobic, yeast biomass production vessel (2 L volume; Candida ingens inoculum; batch [pellicle] growth; 2 day retention time) where the VFAs and some NH4(+)-N were converted into yeast biomass. Yeast yields accounted for up to 8% of potato residue fed into the anaerobic bioreactor. The third bioreactor (0.5 L liquid working volume; commercial nitrifier inoculum; packed-bed biofilm; continuous yeast effluent feed; recirculating; constant volume; 23 day hydraulic retention time) was used to convert successfully the remaining NH4(+)-N into nitrate-N (preferred form of N for CELSS crop production) and to remove the remaining degradable soluble organic carbon. Effluents from the last two stages were used for partial replenishment of minerals for hydroponic potato production.     

Progress in ultrasonic bioreactors for CELSS applications.

An important issue in Controlled Ecological Life Support Systems (CELSS) is the recycling of inedible crop residues to recover inorganic plant nutrients such as nitrates, phosphates, potassium and other macro- and micro-nutrients. In a closed system in space, such regeneration is vital to the long term viability of plant growth necessary for the food production and waste handling process. Chemical approaches to recycling such as incineration and wet oxidation are not compatible with low energy and environmentally friendly regeneration of such nutrients. Biological regeneration is more acceptable environmentally, but it is a very slow process and does not typically result in complete recovery of inorganic and organic nutrients. A new approach to biological regeneration is described here involving the combined use of special enzymatic catalysts and ultrasonic energy in a bioreactor system. This new system has the potential for rapid, efficient, environmentally friendly and complete conversion of crop wastes to inorganic plant nutrients and food recovery from cellulose materials. A series of experimental tests were carried out with a soybean crop residue meal substrate. Biochemical conversion rates were significantly expedited with the addition of enzymes and further enhanced through ultrasonic stimulation of these enzymes. The difference in conversion rates was particularly increased after the initial period of soluble organics conversion. The remaining cellulose substrate is much more difficult to biodegrade, and the ultrasonically-enhanced reaction was able to demonstrate a much higher rate of substrate conversion.     

Integrating biological treatment of crop residue into a hydroponic sweetpotato culture.

Residual biomass from hydroponic culture of sweetpotato [Ipomoea batatas (L.) Lam.] was degraded using natural bacterial soil isolates. Sweetpotato was grown for 120 days in hydroponic culture with a nutrient solution comprised of a ratio of 80% modified half Hoagland solution to 20% filtered effluent from an aerobic starch hydrolysis bioreactor. The phytotoxicity of the effluent was assayed with Waldmann's Green' lettuce (Lactuca sativa L.) and the ratio selected after a 60-day bioassay using sweetpotato plants propagated vegetatively from cuttings. Controlled environment chamber experiments were conducted to investigate the impact of filtrate from biological treatment of crop residue on growth and storage root production with plants grown in a modified half Hoagland solution. Incorporation of bioreactor effluent, reduced storage root yield of 'Georgia Jet' sweetpotato but the decrease was not statistically significant when compared with yield for plants cultured in a modified half Hoagland solution without filtrate. However, yield of 'TU-82-155' sweetpotato was significantly reduced when grown in a modified half Hoagland solution into which filtered effluent had been incorporated. Total biomass was significantly reduced for both sweetpotato cultivars when grown in bioreactor effluent. The leaf area and dry matter accumulation were significantly (P < 0.05) reduced for both cultivars when grown in solution culture containing 20% filtered effluent.     

The effects of composting on the nutritional composition of fibrous bio-regenerative life support systems (BLSS) plant waste residues and its impact on the growth of Nile tilapia (Oreochromis niloticus)

Utilization of bio-regenerative life support systems (BLSS) plant waste residues as a nutritional source by Nile tilapia (Oreochromis niloticus) has proven problematic as a result of high concentrations of fibrous compounds in the plant waste residues. Nutritional improvement of plant waste residues by composting with the oyster mushroom (Pleurotus ostreatus), and the effects on growth and nutrient utilization of Nile tilapia fed such residues were evaluated. Five Nile tilapia (mean weight = 70.9 ± 3.1 g) were stocked in triplicate aquaria and fed one of two experimental diets, cowpea (CP) and composted cowpea (CCP), twice daily for a period of 8 weeks. Composting of cowpea residue resulted in reduced concentrations of nitrogen-free extract, hemi-cellulose and trypsin inhibitor activity, though trypsin inhibitor activity remained high. Composting did not reduce crude fiber, lignin, or cellulose concentrations in the diet. No significant differences (P < 0.05) were observed in weight gain, specific growth rate, survival rate, daily consumption, and food conversion ratio between tilapia fed CP and CCP. These results suggest that P. ostreatus is not a suitable candidate for culture in conjunction with the culture of Nile tilapia. Additional work is needed to determine what, if any, benefit can be obtained from incorporating composted residue as feed for Nile tilapia.     

Impact of emissions from anthropogenic sources on satellite-derived reflectance

The present study deals with the impact of extensive anthropogenic activities associated with festivities and agricultural crop residues burning in the Indo-Gangetic Plains (IGP) on satellite-derived reflectance during November 2007. Intense smoke plumes were observed in the IRS-P4 OCM satellite data over IGP associated with agricultural crop residue burning during the study period. Terra-MODIS AOD and CALIPSO LIDAR backscatter datasets were analysed over the region to understand the spatial and temporal variation of the aerosol properties. Ground-based measurements on aerosol optical properties and black carbon (BC) mass concentration were carried out during 7–14 November 2007 over urban region of Hyderabad, India. Top of the Atmosphere (TOA) reflectance estimated from IRS-P4 Ocean Color Monitor (OCM) data showed large variations due to anthropogenic activities associated with crop residue burning and fireworks. Atmospheric corrections to OCM satellite data using 6S radiative transfer code with inputs from ground and satellite measurements could account for the variations due to differential aerosol loading. Results are discussed in this paper.     

Effects of clinorotation on the polysaccharide content of resynthesized walls of protoplasts.

Changes in cellulose and callose content during cell wall regeneration in Brassica oleracea protoplasts have been examined by cytofluorimetry following their exposure to the conditions of the horizontal clinostat (2 r.p.m.) for 10 days. In comparison with controls, cellulose content decreased 4-fold and 28% of the protoplasts failed to resynthesize a wall in the clinorotated sample. The callose content was almost doubled in clinostated cells. Callose synthesis fluctuated in both control and clinorotated protoplasts. The results support the idea that inhibition of cellulose synthesis in protoplasts grown on the clinostat is caused by a change of plasmalemma fluidity and functioning, and also by a disturbance to the state of cytoplasmic calcium under conditions of simulated microgravity.     

Technical review of the Laboratory Biosphere closed ecological system facility.

Laboratory Biosphere is a 40 m3 closed life system that commenced operation in May 2002. Light is from 12,000 W of high pressure sodium lamps over planting beds with 5.37 m2 of soil. Water is 100% recycled by collecting condensate from the temperature and humidity control system and mixing with leachate collected from under the planting beds. Atmospheric leakage was estimated during the first closure experiment to be 0.5-1% per day in general plus about 1% for each usage of the airlock door. The first trial run of 94 days was with a soybean crop grown from seeds (May 17, 2002) to harvest (August 14, 2002) plus 5 days of post-harvest closure. The focus of this initial trial was system testing to confirm functionality and identify any necessary modifications or improvements. This paper describes the organizational and physical features of the Laboratory Biosphere.     

Recycling of inorganic nutrients for hydroponic crop production following incineration of inedible biomass.

The goal of resource recovery in a regenerative life support system is maintenance of product quality to sure support of reliable and predictable levels of life support function performance by the crop plant component. Further, these systems must be maintained over extended periods of time, requiring maintenance of nutrient solutions to avoid toxicity and deficiencies. The focus of this study was to determine the suitability of the ash product following incineration of inedible biomass as a source of inorganic nutrients for hydroponic crop production. Inedible wheat biomass was incinerated and ash quality characterized. The incinerator ash was dissolved in adequate nitric acid to establish a consistent nitrogen concentration is all nutrient solution treatments. Four experimental nutrient treatments were included: control, ash only, ash supplemented to match the control treatment, and ash only quality formulated with reagent grade chemicals. When nutrient solutions were formulated using only ash following incineration of inedible biomass, a balance in solution is established representing elemental retention following incineration and nutrient proportions present in the original biomass. The resulting solution is not identical to the control. This imbalance resulted in a suppression of crop growth. When the ash is supplemented with reagent grade chemicals to establish the same balance as in the control--growth is identical to the control. The ash appears to carry no phytotoxic materials. Growth in solution formulated with reagent grade chemicals but matching the quality of the ash only treatment resulted in similar growth to that of the ash only treatment. The ash product resulting from incineration of inedible biomass appears to be a suitable form for recycle of inorganic nutrients to crop production.     

The effect of solar–geomagnetic activity during hospital admission on coronary events within 1 year in patients with acute coronary syndromes

Some evidence indicates the deterioration of the cardiovascular system during space storms. It is plausible that the space weather conditions during and after hospital admission may affect the risk of coronary events in patients with acute coronary syndromes (ACS). We analyzed the data of 1400 ACS patients who were admitted to the Hospital Lithuanian University of Health Sciences, and who survived for more than 4 days. We evaluated the associations between geomagnetic storms (GS), solar proton events (SPE), and solar flares (SF) that occurred 0–3 days before and after hospital admission and the risk of cardiovascular death (CAD), non-fatal ACS, and coronary artery bypass grafting (CABG) during a period of 1 year; the evaluation was based on the multivariate logistic model, controlling for clinical data. After adjustment for clinical variables, GS occurring in conjunction with SF 1 day before admission increased the risk of CAD by over 2.5 times. GS 2 days after SPE occurred 1 day after admission increased the risk of CAD and CABG by over 2.8 times. The risk of CABG increased by over 2 times in patients admitted during the day of GS and 1 day after SPE. The risk of ACS was by over 1.63 times higher for patients admitted 1 day before or after solar flares.     

Optimization of moisture content for wheat seedling germination in a cellulose acetate medium for a space flight experiment.

The Porous Tube Plant Nutrient Delivery System (PTPNDS), a hydrophilic, microporous ceramic tube hydroponic system designed for microgravity, will be tested in a middeck locker of the Space Shuttle. The flight experiment will focus on hardware operation and assess its ability to support seed germination and early seedling growth in microgravity. The water controlling system of the PTPNDS hardware has been successfully tested during the parabolic flight of the KC-135. One challenge to the development of the space flight experiment was to devise a method of holding seeds to the cylindrical porous tube. The seed-holder must provide water and air to the seed, absorb water from the porous tube, withstand sterilization, provide a clear path for shoots and roots to emerge, and be composed of flight qualified materials. In preparation for the flight experiment, a wheat seed-holder has been designed that utilizes a cellulose acetate plug to facilitate imbibition and to hold the wheat seeds in contact with the porous tube in the correct orientation during the vibration of launch and the microgravity environment of orbit. Germination and growth studies with wheat at a range of temperatures showed that optimal moisture was 78% (by weight) in the cellulose acetate seed holders. These and other design considerations are discussed.     

Characteristics of the soil-like substrates produced with a novel technique combining aerobic fermentation and earthworm treatment

The soil-like substrate (SLS) technique is key for improving the closure of bioregenerative life support system (BLSS) by recycling the inedible biomass of higher plants. In this study, a novel SLS technique (NSLST) was proposed: aerobic fermentations at 35 °C for 1 day, then 60 °C for 6 days, finally 30 °C for 3 days, followed by earthworm treatment for 70 days. Comparing with the original SLS technique (OSLST), its process cycle was 13 days shorter, and the dry weight loss rate (81.1%) was improved by 24.77%. The cellulose and lignin degradation rates were 96.6% and 94.6%. The concentrations of available N, P and K in mature SLS were respectively 776.1 mg/L, 348.0 mg/L and 7943.0 mg/L. Low CH₄ and NH₃ production was observed, but no accumulation. According to the seed germination test, the SLSs were feasible for plant growth. This investigation will provide a preliminary foundation for BLSS design.     

Atmospheric dynamics in the "Laboratory Biosphere" with wheat and sweet potato crops.

Laboratory Biosphere is a 40-m3 closed life system equipped with 12,000 W of high pressure sodium lamps over planting beds with 5.37 m2 of soil. Atmospheric composition changes due to photosynthetic fixation of carbon dioxide and corresponding production of oxygen or the reverse, respiration, are observed in short timeframes, e.g., hourly. To focus on inherent characteristics of the crop as distinct from its area or the volume of the chamber, we report fixation and respiration rates in mmol h-1 m-2 of planted area. An 85-day crop of USU Apogee wheat under a 16-h lighted/8-h dark regime peaked in fixation rate at about 100 mmol h-1 m-2 approximately 24 days after planting. Light intensity was about 840 micromoles m-2 s-1. Dark respiration peaked at about 31 mmol h-1 m-2 at the same time. Thereafter, both fixation and respiration declined toward zero as harvest time approached. A residual soil respiration rate of about 1.9 mmol h-1 m-2 was observed in the dark closed chamber for 100 days after the harvest. A 126-day crop of Tuskegee TU-82-155 sweet potato behaved quite differently. Under a 680 micromoles m-2 s-1, 18-h lighted/6-h dark regime, fixation during lighted hours rose to a plateau ranging from about 27 to 48 mmol h-1 m-2 after 42 days and dark respiration settled into a range of 12-23 mmol h-1 m-2. These rates continued unabated until the harvest at 126 days, suggesting that tuber biomass production might have continued at about the same rate for some time beyond the harvest time that was exercised in this experiment. In both experiments CO2 levels were allowed to range widely from a few hundred to about 3000 ppm, which permitted observation of fixation rates both at varying CO2 concentrations and at each number of days after planting. This enables plotting the fixation rate as a function of both variables. Understanding the atmospheric dynamics of individual crops will be essential for design and atmospheric management of more complex CELSS which integrate the simultaneous growth of several crops as in a sustainable remote life support system.     

Crop yield and light/energy efficiency in a closed ecological system: Laboratory Biosphere experiments with wheat and sweet potato.

Two crop growth experiments in the soil-based closed ecological facility, Laboratory Biosphere, were conducted from 2003 to 2004 with candidate space life support crops. Apogee wheat (Utah State University variety) was grown, planted at two densities, 400 and 800 seeds m-2. The lighting regime for the wheat crop was 16 h of light-8 h dark at a total light intensity of around 840 micromoles m-2 s-1 and 48.4 mol m-2 d-1 over 84 days. Average biomass was 1395 g m-2, 16.0 g m-2 d-1 and average seed production was 689 g m-2 and 7.9 g m-2 d-1. The less densely planted side was more productive than the denser planting, with 1634 g m-2 and 18.8 g m-2 d-1 of biomass vs. 1156 g m-2 and 13.3 g m-2 d-1; and a seed harvest of 812.3 g m-2 and 9.3 g m-2 d-1 vs. 566.5 g m-2 and 6.5 g m-2 d-1. Harvest index was 0.49 for the wheat crop. The experiment with sweet potato used TU-82-155 a compact variety developed at Tuskegee University. Light during the sweet potato experiment, on a 18 h on/6 h dark cycle, totaled 5568 total moles of light per square meter in 126 days for the sweet potatoes, or an average of 44.2 mol m-2 d-1. Temperature regime was 28 +/- 3 degrees C day/22 +/- 4 degrees C night. Sweet potato tuber yield was 39.7 kg wet weight, or an average of 7.4 kg m-2, and 7.7 kg dry weight of tubers since dry weight was about 18.6% wet weight. Average per day production was 58.7 g m-2 d-1 wet weight and 11.3 g m-2 d-1. For the wheat, average light efficiency was 0.34 g biomass per mole, and 0.17 g seed per mole. The best area of wheat had an efficiency of light utilization of 0.51 g biomass per mole and 0.22 g seed per mole. For the sweet potato crop, light efficiency per tuber wet weight was 1.33 g mol-1 and 0.34 g dry weight of tuber per mole of light. The best area of tuber production had 1.77 g mol-1 wet weight and 0.34 g mol-1 of light dry weight. The Laboratory Biosphere experiment's light efficiency was somewhat higher than the USU field results but somewhat below greenhouse trials at comparable light levels, and the best portion of the crop at 0.22 g mol-1 was in-between those values. Sweet potato production was overall close to 50% higher than trials using hydroponic methods with TU-82-155 at NASA JSC. Compared to projected yields for the Mars on Earth life support system, these wheat yields were about 15% higher, and the sweet potato yields averaged over 80% higher.     

Earth benefits of interdisciplinary CELSS-related research by the NSCORT in Bioregenerative Life Support.

Earth benefits of research from the NSCORT in Bioregenerative Life Support will include the following: development of active control mechanisms for light, CO2, and temperature to maximize photosynthesis of crop plants during important phases of crop development; automation of crop culture systems; creation of novel culture systems for optimum productivity; creation of value-added crops with superior nutritional, yield, and waste-process characteristics; environmental control of food and toxicant composition of crops; new process technologies and novel food products for safe, nutritious, palatable vegetarian diets; creation of menus for healthful vegetarian diets with psychological acceptability; enzymatic procedures to degrade recalcitrant crop residues occurring in municipal waste; control-system strategies to ensure sustainabilty of a CELSS that will enable management of diverse complex systems on Earth.     

Differentiation of cartilaginous anlagen in entire embryonic mouse limbs cultured in a rotating bioreactor.

Mechanisms involved in development of the embryonic limb have remained the same throughout eons of genetic and environmental evolution under Earth gravity (1 g). During the spaceflight era it has been of interest to explore the ancient theory that form of the skeleton develops in response to gravity, and that changes in gravitational forces can change the developmental pattern of the limb. This has been shown in vivo and in vitro, allowing the hypergravity of centrifugation and microgravity of space to be used as tools to increase our knowledge of limb development. In recapitulations of spaceflight experiments, premetatarsals were cultured in suspension in a bioreactor, and found to be shorter and less differentiated than those cultured in standard culture dishes. This study only measured length of the metatarsals, and did not account for possible changes due to the skeletal elements having a more in vivo 3D shape while in suspension vs. flattened tissues compressed by their own weight. A culture system with an outcome closer to in vivo and that supports growth of younger limb buds than traditional systems will allow studies of early Hox gene expression, and contribute to the understanding of very early stages of development. The purpose of the current experiment was to determine if entire limb buds could be cultured in the bioreactor, and to compare the growth and differentiation with that of culturing in a culture dish system. Fore and hind limbs from E11-E13 ICR mouse embryos were cultured for six days, either in the bioreactor or in center-well organ culture dishes, fixed, and embedded for histology. E13 specimens grown in culture dishes were flat, while bioreactor culture specimens had a more in vivo-like 3D limb shape. Sections showed excellent cartilage differentiation in both culture systems, with more cell maturation, and hypertrophy in the specimens cultured in the bioreactor. Younger limb buds fused together during culture, so an additional set of E11.5 limb buds was cultured with and without encapsulation in alginate prior to culturing in the bioreactor. Encapsulated limbs grown in the bioreactor did not fuse together, but developed only the more proximal elements while limbs grown in culture dishes formed proximal and distal elements. Alginate encapsulation may have reduced oxygenation to the progress zone of the developing limb bud resulting in lack of development of the more distal elements. These results show that the bioreactor supports growth and differentiation of skeletal elements in entire E13 limb buds, and that a method to culture younger limb buds without fusing together needs to be developed if any morphometric analysis is to be performed.     

Three-dimensional transgenic cell model to quantify genotoxic effects of space environment.

In this paper we describe a three-dimensional, multicellular tissue-equivalent model, produced in NASA-designed, rotating wall bioreactors using mammalian cells engineered for genomic containment of multiple copies of defined target genes for genotoxic assessment. Rat 2 lambda fibroblasts, genetically engineered to contain high-density target genes for mutagenesis (Stratagene, Inc., Austin, TX), were cocultured with human epithelial cells on Cytodex beads in the High Aspect Ratio Bioreactor (Synthecon, Inc, Houston, TX). Multi-bead aggregates were formed by day 5 following the complete covering of the beads by fibroblasts. Cellular retraction occurred 8-14 days after coculture initiation culminating in spheroids retaining few or no beads. Analysis of the resulting tissue assemblies revealed: multicellular spheroids, fibroblasts synthesized collagen, and cell viability was retained for the 30-day test period after removal from the bioreactor. Quantification of mutation at the LacI gene in Rat 2 lambda fibroblasts in spheroids exposed to 0-2 Gy neon using the Big Blue color assay (Stratagene, Inc.), revealed a linear dose-response for mutation induction. Limited sequencing analysis of mutant clones from 0.25 or 1 Gy exposures revealed a higher frequency of deletions and multiple base sequencing changes with increasing dose. These results suggest that the three-dimensional, multicellular tissue assembly model produced in NASA bioreactors are applicable to a wide variety of studies involving the quantification and identification of genotoxicity including measurement of the inherent damage incurred in Space.     

Controlled Ecological Life Support Systems (CELSS) flight experimentation.

The NASA CELSS program has the goal of developing life support systems for humans in space based on the use of higher plants. The program has supported research at universities with a primary focus of increasing the productivity of candidate crop plants. To understand the effects of the space environment on plant productivity, the CELSS Test Facility (CTF) has been been conceived as an instrument that will permit the evaluation of plant productivity on Space Station Freedom. The CTF will maintain specific environmental conditions and collect data on gas exchange rates and biomass accumulation over the growth period of several crop plants grown sequentially from seed to harvest. The science requirements of the CTF will be described, as will current design concepts and specific technology requirements for operation in micro-gravity.     

Integration of waste processing and biomass production systems as part of the KSC Breadboard project.

After initial emphasis on large-scale baseline crop tests, the Kennedy Space Center (KSC) Breadboard project has begun to evaluate long-term operation of the biomass production system with increasing material closure. Our goal is to define the minimum biological processing necessary to make waste streams compatible with plant growth in hydroponic systems, thereby recycling nutrients into plant biomass and recovering water via atmospheric condensate. Initial small and intermediate-scale studies focused on the recycling of nutrients contained in inedible plant biomass. Studies conducted between 1989-1992 indicated that the majority of nutrients could be rapidly solubilized in water, but the direct use of this crop "leachate" was deleterious to plant growth due to the presence of soluble organic compounds. Subsequent studies at both the intermediate scale and in the large-scale Biomass Production Chamber (BPC) have indicated that aerobic microbiological processing of crop residue prior to incorporation into recirculating hydroponic solutions eliminated any phytotoxic effect, even when the majority of the plant nutrient demand was provided from recycled biomass during long term studies (i.e. up to 418 days). Current and future studies are focused on optimizing biological processing of both plant and human waste streams.     

Incineration of biomass and utilization of product gas as a CO2 source for crop production in closed systems: gas quality and phytotoxicity.

This study addressed the recycle of carbon from inedible biomass to CO2 for utilization in crop production. Earlier work identified incineration as an attractive approach to resource recovery from solid wastes because the products are well segregated. Given the effective separation of carbon into the gaseous product stream from the incinerator in the form of CO2 we captured the gaseous stream produced during incineration of wheat inedible biomass and utilized it as the CO2 source for crop production. Injection rate was based on maintenance of CO2 concentration in the growing environment. The crop grown in the closed system was lettuce. Carbon was primarily in the form of CO2 in the incinerator product gas with less than 8% of carbon compounds appearing as CO. Nitrogen oxides and organic compounds such as toluene, xylene, and benzene were present in the product gas at lower concentrations (< 4 micromol mol-1); sulfur containing compounds were below the detection limits. Direct utilization of the gaseous product of the incinerator as the CO2 source was toxic to lettuce grown in a closed chamber. Net photosynthetic rates of the crop was suppressed more than 50% and visual injury symptoms were visible within 3 days of the introduction of the incinerator gas. Even the removal of the incinerator gas alter two days of crop exposure and replacement with pure CO2 did not eliminate the toxic effects. Both organic and inorganic components of the incinerator gas are candidates for the toxin.     

Search for heterocyclic radiolytic products in aqueous solutions of cyanides

This work is a part of a systematic search for biologically significant compounds which are formed during radiolysis of aqueous solutions of cyano compounds. Aqueous solutions of HCN (0.2M, O₂-free, natural pH) were irradiated at 18 Mrad dose of ⁶⁰Co gammas, and the non-volatile radiolytic products concentrated in a dry residue. An aliquot of the dry residue was hydrolyzed with formic acid and the hydrolyzate selectively absorbed on active charcoal. The bulk of products from the charcoal was passed through a SP Sephadex C-25 column and the fractions analyzed by means of high performance liquid chromatography on a reverse phase column. Tentative assignments are based on retention times in the HPLC procedure and their pH dependence, and the coinjection of standards. The presence of adenine is certain. There is supporting evidence for uracil, thymine and cytosine. Experimental results also strongly support the presence of triazines.