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NASA NTRSDate not supplied

Update to Developing a Multi-bioreactor Test Stand for the Evaluation of Biofilm Mitigation Technologies

Biofilm formation in spacecraft wastewater systems is a complex and impactful problem. Biomass flowing in these wastewater systems can cause clogging, higher differential pressures, and flow restrictions impacting equipment reliability and function. To explore this issue, the Marshall Space Flight Center’s biofilm test stand was created and has been utilized to examine multiple biofilm mitigation techniques over a long duration test time. This paper is an update to a previous paper, ICES-2021-122, and will serve to discuss the modifications and enhancements to the biofilm test stand. While the previous configuration tested heat treatments and coatings, this phase of testing will examine differing materials, competing bacteria provided by an external bioreactor, a prototype phytoremediation unit, and additional testing with a nutrient filter. Configuration changes include the addition of float switches, adjustment of the drain assembly architecture and additional support hardware for the competing bacteria bioreactor and phytoremediation units.

ECLSS↗

NASA NTRSDate not supplied

Update to Developing a Multi-bioreactor Test Stand for the Evaluation of Biofilm Mitigation Technologies

ECLSS↗

NASA NTRS1984

Tomato growth as affected by root-zone temperature and the addition of gibberellic acid and kinetin to nutrient solutions

The effect of root-zone temperature on young tomato plants (Lycopersicon esculentum Mill. cv. Heinz 1350) was evaluated in controlled environments using a recirculating solution culture system. Growth rates were measured at root-zone temperatures of 15 degrees, 20 degrees, 25 degrees, and 30 degrees C in a near optimum foliar environment. Optimum growth occurred at 25 degrees to 30 degrees during the first 4 weeks of growth and 20 degrees to 25 degrees during the 5th and 6th weeks. Growth was severely restricted at 15 degrees. Four concentrations of gibberellic acid (GA3) and kinetin were added to the nutrient solution in a separate trial; root-zone temperature was maintained at 15 degrees and 25 degrees. Addition of 15 micromoles GA3 to solutions increased specific leaf area, total leaf area, and dry weight production of plants in both temperature treatments. GA3-induced growth stimulation was greater at 15 degrees than at 25 degrees. GA3 may promote growth by increasing leaf area, enhancing photosynthesis per unit leaf area, or both. Kinetic was not useful in promoting growth at either temperature.

NASA Program CELSS↗

DOE OSTI2020

Microbial processing of plant remains is co-limited by multiple nutrients in global grasslands

Microbial processing of aggregate-unprotected organic matter inputs is key for soil fertility, long-term ecosystem carbon and nutrient sequestration and sustainable agriculture. In this work, we investigated the effects of adding multiple nutrients (nitrogen, phosphorus and potassium plus nine essential macro- and micro-nutrients) on decomposition and biochemical transformation of standard plant materials buried in 21 grasslands from four continents. Addition of multiple nutrients weakly but consistently increased decomposition and biochemical transformation of plant remains during the peak-season, concurrent with changes in microbial exoenzymatic activity. Higher mean annual precipitation and lower mean annual temperature were the main climatic drivers of higher decomposition rates, while biochemical transformation of plant remains was negatively related to temperature of the wettest quarter. Nutrients enhanced decomposition most at cool, high rainfall sites, indicating that in a warmer and drier future fertilized grassland soils will have an even more limited potential for microbial processing of plant remains.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2025

Temporal Galactose‐Manganese Feeding in Fed‐Batch and Perfusion Bioreactors Modulates UDP‐Galactose Pools for Enhanced mAb Glycosylation Homogeneity

ABSTRACT Monoclonal antibodies (mAbs) represent a majority of biotherapeutics in the market today. These glycoproteins undergo posttranslational modifications, such as N‐linked glycosylation, that influence the structural & functional characteristics of the antibody. Glycosylation is a heterogenous posttranslational modification that may influence therapeutic glycoprotein stability and clinical efficacy, which is why it is often considered a critical quality attribute (CQA) of the mAb product. While much is known about the glycosylation pathways of Chinese Hamster Ovary (CHO) cells and how cell culture chemical modifiers may influence the N‐glycosylation profile of the final product, this knowledge is often based on the final cumulative glycan profile at the end of the batch process. Building a temporal understanding of N‐glycosylation and how mAb glycoform composition responds to real‐time changes in the biomanufacturing process will help build integrated process models that may allow for glycosylation control to produce a more homogenous product. Here, we look at the effect of specific nutrient feed media additives (e.g., galactose, manganese) and feeding times on the N‐glycosylation pathway to modulate N‐glycosylation of a Herceptin biosimilar mAb (i.e., Trastuzumab). We deploy the N‐GLYcanyzer process analytical technology (PAT) to monitor glycoforms in near real‐time for bench‐scale bioprocesses operated in both fed‐batch and perfusion modes to build an understanding of how temporal changes in mAb N‐glycosylation are dependent on specific media additives. We find that Trastuzumab terminal galactosylation is sensitive to media feeding times and intracellular nucleotide sugar pools. Temporal analysis reveals an increased desirable production of single and double galactose‐occupied glycoforms over time under glucose‐starved fed‐batch cultures. Comparable galactosylation profiles were also observed between fed‐batch (nutrient‐limited) and perfusion (non‐nutrient‐limited) bioprocess conditions. In summary, our results demonstrate the utility of real‐time monitoring of mAb glycoforms and feeding critical cell culture nutrients under fed‐batch and perfusion bioprocessing conditions to produce higher‐quality biologics.

Biotechnology & Applied Microbiology↗

NASA NTRS1999

Fe(VI) as a Possible Oxidant on the Martian Surface

The essential findings of the three biological experiments (Gas Exchange, Labeled Released, and Pyrolitic Release) aboard the Viking Mars landers were the discovery of the presence of one or more strong oxidants on the Martian surface. The Gas Exchange experiments showed that wetting Martian soil leads to the evolution of oxygen, while in the Labeled Release experiment addition of a nutrient solution containing C-14-labeled formate, glycine, lactate, alanine, and glycolic acid induced CO2 evolution. A general consensus was reached that all data taken together pointed to the presence on Martian surface of a strong oxidant, or most probably several different types of oxidants. Several candidates have been proposed as oxidants, including superoxides, hydrogen peroxide, and iron oxides (possibly gamma-Fe2O3). Additional information is contained in the original extended abstract.

Tsapin, A. I.↗

NASA NTRSDate not supplied

Testing Specialized Fertigation Blends for Agronomic Biofortification for Spaceflight Applications

The current spaceflight diet is prepackaged and vitamins degrade over time. Plants grown in the Veggie and APH units on the ISS currently provide the astronauts with a hefty dose of vitamins and minerals, but some nutrients could be improved. Agronomic Biofortifcation of crops with fertilizers has been tested on Earth, with the successful addition of minerals and vitamin C to crops that have otherwise been deficient. We intend to test if we can replicate this in a controlled environment and expand it to nutrients that are not often present in abundance in plants. This will be done by adding fluid nutrient blends to the water supply of candidate crops for spaceflight applications. We hypothesize that fertilization of plants with nutrients that are of value to human health will have neutral to positive effect on plants, with desired nutrients present in greater abundance in treated plants than untreated plants. We intend to show this through germination percent and pace, yield of edible biomass, the growth rate, and overall development of the crop plant. For this experiment, radish microgreens were grown on mats in a controlled environment chamber in enriched CO2, with LED lighting similar to what is found in the ISS Veggie hardware. Watering was conducted once at planting with a specialized blend of ½ strength Hoagland’s hydroponic solution with additional vitamins. Preliminary results show no decrease in overall biomass production with plants grown using additional vitamin D, but additional nutrients will be tested. Future direction will be to perform nutritional analysis of plants. Test that resulting produce will be suitable for human consumption, including meeting or exceeding food safety guidelines and palatability.

Christina M Johnson↗

NASA NTRSDate not supplied

Testing Specialized Fertigation Blends for Agronomic Biofortification for Spaceflight Applications

microgreens↗

DOE OSTI2025

From subsidies to stressors: Positively skewed ecological gradients alter biological responses to nutrients in streams

Abstract Subsidy–stress gradients offer a useful framework for understanding ecological responses to perturbation and may help inform ecological metrics in highly modified systems. Historic, region‐wide shifts from bottomland hardwood forest to row crop agriculture can cause positively skewed impact gradients in alluvial plain ecoregions, resulting in tolerant organisms that typically exhibit a subsidy response (increased abundance in response to environmental stressors) shifting to a stress response (declining abundance at higher concentrations). As a result, observed biological tolerance in modified ecosystems may differ from less modified regions, creating significant challenges for detecting biological responses to restoration efforts. Using the agriculturally dominated Mississippi Alluvial Plain (MAP) ecoregion in Mississippi, USA, as a case study, we tested the hypothesis that macroinvertebrate taxa that typically display a subsidy response to nutrient enrichment in less modified ecoregions (i.e., nutrient‐tolerance) shift to a stress response to increasing nutrients in highly modified watersheds with elevated baseline nutrient conditions (i.e., nutrient intolerance). The abundance and diversity of MAP‐specific intolerant taxa identified with threshold indicator taxa analysis were either unresponsive or exhibited a subsidy response to increasing nutrients in less modified ecoregions in Mississippi with less land alteration and lower nutrient concentrations, but declined at higher concentrations, providing evidence for a stress response to elevated nutrients in the MAP. Additionally, MAP‐specific tolerant and intolerant taxa richness responded to increased nutrients predictably and consistently across space and time within the MAP. However, in MAP streams, elevated specific conductance was predicted to dampen the response of tolerant and intolerant taxa richness to increasing nutrient concentrations, highlighting the importance of considering multistressor interactions when interpreting biological data. Lastly, we demonstrate the efficacy of this approach with sediment bacterial communities characterized with amplicon sequencing, which lack sufficient life history characteristics necessary for the development of multimetric indices. Both macroinvertebrate and bacterial communities responded similarly to increasing nutrient concentrations, suggesting DNA‐based approaches may provide an efficient biological assessment tool for monitoring water quality improvements in highly modified watersheds.

DeVilbiss, Stephen E. [U.S. Geological Survey Lowe↗

DOE OSTI2021

Conversion of High-Solids Hydrothermally Pretreated Bioenergy Sorghum to Lipids and Ethanol Using Yeast Cultures

Glucose and xylose are the major sugars in cellulosic hydrolysates. The cellulosic sugars can be used for the production of biofuels and value-added bioproducts. In this study, lipid and ethanol were produced from bioenergy sorghum syrups using engineered yeasts. Here, bioenergy sorghum was hydrothermally pretreated at 50% solids loading in a continuous reactor system and mechanically refined sequentially using a burr mill to improve biomass accessibility for hydrolysis. Fed-batch enzymatic hydrolysis was conducted with 50% w/v solids loading to achieve 230 g/L sugar concentration. Different strains of Rhodosporidium toruloides were used to ferment sugars into lipids, and the highest lipid yield of 9.2 g/L was observed. The lipid yield was improved to 19.0 g/l by implementing a two-stage culture where once the sugars were exhausted in the first stage, the yeast was introduced into fresh hydrolysate without adding nitrogen. For ethanol production, the engineered Saccharomyces cerevisiae SR8ΔADH6 was utilized to coferment glucose and xylose. Additionally, the effects of nutrient media (YP, YNB/urea, and urea), cellulosic sugar concentration, and sulfite addition were investigated to optimize the ethanol yield from sorghum syrups. The optimal ethanol yield at 73.3% was obtained from the YNB/urea culture consisting of 34 g glucose/L and 17 g xylose/L.

09 BIOMASS FUELS↗

DOE OSTI2025

Scale-up study of enhancing algal growth through bioaugmentation with the indole-3-acetic acid producing bacteria Azospirillum brasilense

Algae-based wastewater treatment technologies can recover nutrients while generating algal biomass with diverse application potentials. While numerous bacteria have demonstrated the ability to enhance algal growth through symbiotic relationships in laboratory settings, scaling this benefit to larger applications remains challenging due to complex wastewater conditions and microbe-algae dynamics. This study explores the effectiveness of bioaugmentation with Azospirillum brasilense, a plant growth-promoting bacterium known for producing the phytohormone indole-3-acetic acid (IAA), in boosting algal productivity within an Algaewheel wastewater treatment system. Through a series of experiments conducted at lab-, pilot-, and full-scale levels, we evaluated the impact of bioaugmentation on algal growth and developed a bioaugmentation strategy for an Algaewheel system. The results indicate that a biweekly dosing, achieving a final bioaugmenting cell density of 0.2–4 × 10 9 cells/L, effectively doubles the biomass productivity in a residential subdivision wastewater treatment plant in Northern Illinois. The bacterial production of IAA was the primary mechanism driving this enhancement, as demonstrated by the similar growth and yield improvements observed with chemically supplemented IAA and bioaugmentation across all experimental scales. High-throughput sequencing of 18S rRNA genes revealed that the algal community in the bioaugmented tank exhibited more stable biodiversity than the control. Additionally, bioaugmentation improved nutrient removal efficiency during winter, highlighting its potential to enhance the overall performance and sustainability of algae-based wastewater systems. The developed bioaugmentation and chemical treatment methods offer further operational solutions for managing yield, optimizing biochemical profiles, and enhancing biofuel production potential.

Algae-bacterial symbiotic relationship↗

DOE OSTI2025

Addressing genome scale design tradeoffs in Pseudomonas putida for bioconversion of an aromatic carbon source

Genome-scale metabolic models (GSMM) are commonly used to identify gene deletion sets that result in growth coupling and pairing product formation with substrate utilization and can improve strain performance beyond levels typically accessible using traditional strain engineering approaches. However, sustainable feedstocks pose a challenge due to incomplete high-resolution metabolic data for non-canonical carbon sources required to curate GSMM and identify implementable designs. Here we address a four-gene deletion design in the Pseudomonas putida KT2440 strain for the lignin-derived non-sugar carbon source, p-coumarate (p-CA), that proved challenging to implement. We examine the performance of the fully implemented design for p-coumarate to glutamine, a useful biomanufacturing intermediate. In this study glutamine is then converted to indigoidine, an alternative sustainable pigment and a model heterologous product that is commonly used to colorimetrically quantify glutamine concentration. Through proteomics, promoter-variation, and growth characterization of a fully implemented gene deletion design, we provide evidence that aromatic catabolism in the completed design is rate-limited by fumarase hydratase (FUM) enzyme activity in the citrate cycle and requires careful optimization of another fumarate hydratase protein (PP_0897) expression to achieve growth and production. A double sensitivity analysis also confirmed a strict requirement for fumarate hydratase activity in the strain where all genes in the growth coupling design have been implemented. Metabolic cross-feeding experiments were used to examine the impact of complete removal of the fumarase hydratase reaction and revealed an unanticipated nutrient requirement, suggesting additional functions for this enzyme. While a complete implementation of the design was achieved, this study highlights the challenge of completely inactivating metabolic reactions encoded by under-characterized proteins, especially in the context of multi-gene edits.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS1976

Skylab nutritional studies

Precise nutritional specifications arising from both physiological and experimental requirements necessitated a comprehensive study of the chemical composition of the Skylab food supply. Each of the approximately 70 different food items was analyzed for digestible and nondigestible carbohydrate, and for protein, amino acids, fat, fatty acids, vitamins, and minerals. Menus were formulated to provide at least the National Research Council's Recommended Dietary Allowance of all essential nutrients and, in addition, to provide constant daily intakes of calcium, phosphorus, magnesium, sodium, potassium and protein. In general, the crewmembers adhered to their programmed menus. The ability to swallow and digest food was unaffected by prolonged weightlessness. Taste acuity also appeared to be undiminished inflight. The bone and muscle changes which occurred in previous flights were more pronounced in Skylab. It is concluded that these changes did not develop as a result of nutritional deficit. If such changes are nutritionally related, they point to the existence of nutritional requirements in weightlessness which differ quantitatively from those observed on earth.

Rambaut, P. C.↗

NASA NTRS1977

Skylab nutritional studies

Precise nutritional specifications arising from both physiological and experimental requirements necessitated a comprehensive study of the chemical composition of the Skylab food supply. Each of the approximately seventy different food items was analyzed for digestible and non-digestible carbohydrate, and for protein, amino acids, fat, fatty acids, vitamins and minerals. Menus were formulated to provide at least the National Research Council's Recommended Dietary Allowance of all essential nutrients and, in addition, to provide constant daily intakes of calcium, phosphorus, magnesium, sodium, potassium and protein. In general, the crew members adhered to their programmed menus. The ability to swallow and digest food was unaffected by prolonged weightlessness. Taste acuity also appeared to be undiminished in flight. The bone and muscle changes which occurred in previous flights were more pronounced in Skylab. It is concluded that these changes did not develop as a result of nutritional deficit. If such changes are nutritionally related, they point to the existence of nutritional requirements in weightlessness which differ quantitatively from those observed on earth.

long duration↗

NASA NTRS2009

Expert system for controlling plant growth in a contained environment

In a system for optimizing crop growth, vegetation is cultivated in a contained environment, such as a greenhouse, an underground cavern or other enclosed space. Imaging equipment is positioned within or about the contained environment, to acquire spatially distributed crop growth information, and environmental sensors are provided to acquire data regarding multiple environmental conditions that can affect crop development. Illumination within the contained environment, and the addition of essential nutrients and chemicals are in turn controlled in response to data acquired by the imaging apparatus and environmental sensors, by an ''expert system'' which is trained to analyze and evaluate crop conditions. The expert system controls the spatial and temporal lighting pattern within the contained area, and the timing and allocation of nutrients and chemicals to achieve optimized crop development. A user can access the ''expert system'' remotely, to assess activity within the growth chamber, and can override the ''expert system''.

May, George A.↗

NASA NTRS2011

Expert system for controlling plant growth in a contained environment

In a system for optimizing crop growth, vegetation is cultivated in a contained environment, such as a greenhouse, an underground cavern or other enclosed space. Imaging equipment is positioned within or about the contained environment, to acquire spatially distributed crop growth information, and environmental sensors are provided to acquire data regarding multiple environmental conditions that can affect crop development. Illumination within the contained environment, and the addition of essential nutrients and chemicals are in turn controlled in response to data acquired by the imaging apparatus and environmental sensors, by an "expert system" which is trained to analyze and evaluate crop conditions. The expert system controls the spatial and temporal lighting pattern within the contained area, and the timing and allocation of nutrients and chemicals to achieve optimized crop development. A user can access the "expert system" remotely, to assess activity within the growth chamber, and can override the "expert system".

May, George A.↗

NASA NTRS1998

The Coupling of Solution Chemistry to Plant Nutrient Demand in an on Demand Nutrient Delivery System

The goal of the proposal will be to determine the suitability of the DASI instrument in providing a signal that can be recognized and be utilized as an indicator of plant stress. The method to be utilized for evaluating stress is the presentation of an every increasing level of nutrient deficiency and salinity stress (addition of salt (NACl) or increasing concentration of balanced nutrient) while simultaneously recording spectral reflectance using the DASI instrument and monitoring the traditional processes of gas exchange and nutrient uptake parameters. In this manner, we will be able to directly compare the DASI measurements with known stresses as determined by the traditional gas exchange and nutrient uptake measures of stress. We anticipate that the DASI will provide a sensitive identifier of plant stress; recording signals of the resulting changes in plant metabolism in real time, far before any visible effects of stress could be observed. Thus, there is a potential for very early management intervention to correct a stress condition before damage could develop. The present response time for the observation of visual symptoms of plant stress is considerable and only provides an indication that a stress is present after it has been present for an extended period of time. Thus, the impact of a plant-based life support function will have already been significant. An additional benefit of this research to regenerative life support will be the characterization of a potential recovery scenario from various degrees of stress. The experimental approach to be employed includes the removal of the stress at various points in the stress gradient and the characterization of plant performance and reflectance spectra during recovery from various degrees of stress. Spectral reflectance imaging techniques have been developed and used to measure the biochemical composition of plants and relate these characteristics to the fluxes of biochemical elements within the ecosystem.

Savage, Wayne↗

NASA NTRS1987

Design of an elemental analysis system for CELSS research

The results of experiments conducted with higher plants in tightly sealed growth chambers provide definite evidence that the physical closure of a chamber has significant effects on many aspects of a plant's biology. One of these effects is seen in the change in rates of uptake, distribution, and re-release or nutrient elements by the plant (mass balance). Experimental data indicates that these rates are different from those recorded for plants grown in open field agriculture, or in open growth chambers. Since higher plants are a crucial component of a controlled ecological life support system (CELSS), it is important that the consequences of these rate differences be understood with regard to the growth and yield of the plants. A description of a system for elemental analysis which can be used to monitor the mass balance of nutrient elements in CELSS experiments is given. Additionally, data on the uptake of nutrient elements by higher plants grown in a growth chamber is presented.

Schwartzkopf, Steven H.↗