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At least 1,423 records · Page 79

Continuous multimodal technologies in industrial microbiology: potential for achieving high process performance and agility

The field of industrial microbiology has great potential to tackle the most pressing challenges facing society, but today, this entails much more than classical fermentation technology followed by product upgrading. Methods are needed to confer unit-operation-level functionality and whole-system-level robustness, as well as flexibility. As examples of this concept, we focus here on methodologies often deployed at industrial scale as discrete processes—such as biological, chemical, and electrochemical unit operations—that are also capable of substantial complementarity when combined sequentially and operated continuously. A key and relatively new type of conversion step is made possible by catalyst immobilization methods at both the enzymatic and cellular level. Cases drawn from electro-fermentation for acetoin production and syngas-mediated microbial electrosynthesis of ethanol exemplify the synergy of combined operational approaches. Because modular processes can be treated as a series of reactor elements, considerable flexibility is possible. As discussed, however, not all processes are amenable to this intensification.

09 BIOMASS FUELS↗

Enhanced Characterization of Microorganisms in the Spacecraft Environment

Spacecraft such as the International Space Station (ISS) and the space shuttles are enclosed environments where crewmembers may spend long periods of time. Currently, crewmembers spend approximately a period of 6 months in the ISS. It is known that these prolonged stays in space may result in weakening of the immune system. Therefore, exposure to opportunistic pathogens or high concentrations of environmental microorganisms may compromise the health of the crew. The detection of biocontaminants in spacecraft environments utilizes culture-based methodology, omitting greater than 90% of all microorganisms including pathogens such as Legionella and Cryptosporidium. Culturable bacteria and fungi have been the only allergens studied; the more potent allergens, such as those from dust mites, have never been tested for in spacecraft environments. In addition, no attempts have been made to monitor microbial toxins in spacecrafts. The present study utilized quantitative polymerase chain reaction (QPCR) as a novel approach for monitoring microorganisms in the spacecraft environment. QPCR is a molecular biology technique that does not rely on the physiological state of the organisms for identification, thereby enabling detection of both culturable and non-culturable organisms. In this project, specific molecular primers and probes were utilized for the detection and quantitation of two fungi of concern in indoor environments, Aspergillus fumigatus and Stachybotrys chartarum. These organisms were selected because of the availability of PCR primers and probes, and to establish the sample processing and analysis methodology that may be employed with additional organisms. Purification methods and QPCR assays were optimized for the detection of these organisms in air, surface, and water; and sample processing and analysis protocols were developed. Preliminary validation of these protocols was conducted in the laboratory with air, surface, and water samples seeded with known concentrations of the target organisms. Additional studies were conducted with bulk materials (HEPA filter pleats and particulate found on the filter screen) obtained from the ISS.

Cruz, Patricia↗

Life in the Clouds of Venus? An Experimental Synthetic Biology Approach

The surface of Venus constitutes the most hellish and biologically inhospitable planetary surface in our solar system, boasting a pH of 0, blistering winds that can melt lead, and pressures of 60 atm. However, during the earlier years of the solar system, without the runaway greenhouse effect that has plagued the planet, Venus potentially housed oceans and perhaps even life. There is a possibility that microbes could have retreated into hospitable niches in the atmosphere, as suggested by Carl Sagan as early as 1967 [1]. For example, 50 km above the raging hell of the Venusian surface, exists a relatively temperate environment that might serve as reservoir for life. This astrobiology project seeks to explore life at the extremes and to theorize whether microbial communities could not only survive but also reproduce in the Venusian atmosphere. Specifically, we ask: are aerosols viable microbial environments? But before we can test for life in the clouds, we have to develop a proper reporter to visualize cell growth in situ. For this purpose, we aimed to develop cell-growth dependent reporters to serve as remote biosensors for cell growth. We developed two using the polA promoter, a DNA-replication dependent promoter, and nrd operon promoter, a cell-cycle dependent promoter. Using these cell-growth reporters, the next step is to aerosolize microbes expressing these reporters in a suspension chamber adapted from a Millikan Drop Apparatus to assay reproduction in an aerosolized environment. Better yet is to test the reproduction of microbes in a microgravity regime such as on ISS.Approach: We engineered two cell-cycle dependent genetic reporters. One was the polA promoter which codes for DNA Polymerase I, a gene active in DNA replication [2]. The other was the nrdP. The activation of ribonucleotide reductase reduces ribonucleotides into deoxyribonucleotides and is involved in the bacterial cell cycle [3]. This promoter began activation during the initiation of DNA replication and is cell-cycle dependent [4]. These promoters were fused to a GFP reporter, transformed into E. coli. The constructs were deposited in the iGEM registry as K847210: Escherichia coli DNA-replication dependent polA promoter K847211: Escherichia coli cell-division dependent nrd promoter Results: Our constructs displayed fluorescence when transformed into NEB-5alpha competent cells. While nrdP-E0840 displayed sufficient fluorescence as verified by fluorescent microscopy, the original polAP-E0840 construct (which uses mut3b GFP) exhibited low expression; while fluorescence was visible under the microscope, the signal was too weak for the camera to recognize. The polA promoter was therefore digest-ed with EcoRI and SpeI then ligated into plasmid pNCS containing a RBS, Clover, and a terminator. Clover is a highly engineered green fluorescent protein that exhibits extreme brightness [5] Fluoresence time course data demonsrated that the genes were induced in a cell cycle dependant manner [6]. Our assays via microscopy and the bulk assay shows that our promoters are functional as cell cycle reporters.Conclusions: The application of such tools are widespread and not limited to astrobiology; nrdP could be used to determine doubling times empirically and could possibly extrapolate DNA content from intensity of signals expressed by polAP. However, we are pri-marily interested in its use in astrobiology.

Escherichia↗

Mars 2020: Mission, Science Objectives and Build

If all goes according to plan, in February 2021, NASA will land the Mars 2020 Rover on the surface of Mars. Mars 2020 is the latest in a series of unmanned Martian robotic rover missions that are part of NASA’s Mars Exploration Program, a long-term effort of robotic exploration of the planet. The mission seeks to address high-priority goals for Mars exploration, including answering questions about the potential for past life on Mars. Mars 2020 will look for evidence of habitable conditions on Mars in the ancient past, as well as look for signs of past microbial life itself. The mission also seeks to understanding the geological history and evolution of the planet, and to prepare for future robotic and human exploration. The Mars 2020 spacecraft and rover borrow heavily from the Mars Science Laboratory (MSL) mission and Curiosity rover which landed on Mars in 2012. This reliance on proven technology helps reduce mission risk and cost. Mars 2020 does contain new technology, including a drill for coring samples from Martian rock and soil and a Sample Caching System for gathering, storing and preserving samples for possible future return to Earth. In this paper, we will review the primary goals of the Mars 2020 Mission and look at the reasons for choosing Jezero Crater as the landing site. We will discuss the design and build of the Mars 2020 Spacecraft system and its similarities and differences with Mars Science Laboratory and the Curiosity Rover. We will also review the Mars 2020 Scientific Instrument Suite and their goals. Finally, we will review the Return Sample Contamination Control requirements and the design choices that were made to facilitate meeting these requirements.

Soares, Carlos E.↗

Disentangling direct from indirect relationships in association networks

Networks are vital tools for understanding and modeling interactions in complex systems in science and engineering, and direct and indirect interactions are pervasive in all types of networks. However, quantitatively disentangling direct and indirect relationships in networks remains a formidable task. Here, we present a framework, called iDIRECT (Inference of Direct and Indirect Relationships with Effective Copula-based Transitivity), for quantitatively inferring direct dependencies in association networks. Using copula-based transitivity, iDIRECT eliminates/ameliorates several challenging mathematical problems, including ill-conditioning, self-looping, and interaction strength overflow. With simulation data as benchmark examples, iDIRECT showed high prediction accuracies. Application of iDIRECT to reconstruct gene regulatory networks in Escherichia coli also revealed considerably higher prediction power than the best-performing approaches in the DREAM5 (Dialogue on Reverse Engineering Assessment and Methods project, #5) Network Inference Challenge. In addition, applying iDIRECT to highly diverse grassland soil microbial communities in response to climate warming showed that the iDIRECT-processed networks were significantly different from the original networks, with considerably fewer nodes, links, and connectivity, but higher relative modularity. Further analysis revealed that the iDIRECT-processed network was more complex under warming than the control and more robust to both random and target species removal ( P < 0.001). As a general approach, iDIRECT has great advantages for network inference, and it should be widely applicable to infer direct relationships in association networks across diverse disciplines in science and engineering.

59 BASIC BIOLOGICAL SCIENCES↗

Metagenomics-resolved genomics provides novel insights into chitin turnover, metabolic specialization, and niche partitioning in the octocoral microbiome

Abstract Background The role of bacterial symbionts that populate octocorals (Cnidaria, Octocorallia) is still poorly understood. To shed light on their metabolic capacities, we examined 66 high-quality metagenome-assembled genomes (MAGs) spanning 30 prokaryotic species, retrieved from microbial metagenomes of three octocoral species and seawater. Results Symbionts of healthy octocorals were affiliated with the taxa Endozoicomonadaceae , Candidatus Thioglobaceae , Metamycoplasmataceae , unclassified Pseudomonadales , Rhodobacteraceae , unclassified Alphaproteobacteria and Ca. Rhabdochlamydiaceae . Phylogenomics inference revealed that the Endozoicomonadaceae symbionts uncovered here represent two species of a novel genus unique to temperate octocorals, here denoted Ca. Gorgonimonas eunicellae and Ca. Gorgonimonas leptogorgiae . Their genomes revealed metabolic capacities to thrive under suboxic conditions and high gene copy numbers of serine-threonine protein kinases, type 3-secretion system, type-4 pili, and ankyrin-repeat proteins, suggesting excellent capabilities to colonize, aggregate, and persist inside their host. Contrarily, MAGs obtained from seawater frequently lacked symbiosis-related genes. All Endozoicomonadaceae symbionts harbored endo-chitinase and chitin-binging protein-encoding genes, indicating that they can hydrolyze the most abundant polysaccharide in the oceans. Other symbionts, including Metamycoplasmataceae and Ca. Thioglobaceae , may assimilate the smaller chitin oligosaccharides resulting from chitin breakdown and engage in chitin deacetylation, respectively, suggesting possibilities for substrate cross-feeding and a role for the coral microbiome in overall chitin turnover. We also observed sharp differences in secondary metabolite production potential between symbiotic lineages. Specific Proteobacteria taxa may specialize in chemical defense and guard other symbionts, including Endozoicomonadaceae , which lack such capacity. Conclusion This is the first study to recover MAGs from dominant symbionts of octocorals, including those of so-far unculturable Endozoicomonadaceae , Ca. Thioglobaceae and Metamycoplasmataceae symbionts. We identify a thus-far unanticipated, global role for Endozoicomonadaceae symbionts of corals in the processing of chitin, the most abundant natural polysaccharide in the oceans and major component of the natural zoo- and phytoplankton feed of octocorals. We conclude that niche partitioning, metabolic specialization, and adaptation to low oxygen conditions among prokaryotic symbionts likely contribute to the plasticity and adaptability of the octocoral holobiont in changing marine environments. These findings bear implications not only for our understanding of symbiotic relationships in the marine realm but also for the functioning of benthic ecosystems at large.

59 BASIC BIOLOGICAL SCIENCES↗

Wake Cycle Robustness of the Mars Science Laboratory Flight Software

The Mars Science Laboratory (MSL) is a spacecraft being developed by the Jet Propulsion Laboratory (JPL) for the purpose of in-situ exploration on the surface of Mars. The objective of MSL is to explore and quantitatively assess a local region on the Martian surface as a habitat for microbial life, past or present. This objective will be accomplished through the assessment of the biological potential of at least one target environment, the characterization of the geology and geochemistry of the landing region, an investigation of the planetary process relevant to past habitability, and a characterization of surface radiation. For this purpose, MSL incorporates a total of ten scientific instruments for which functions are to include, among others, atmospheric and descent imaging, chemical composition analysis, and radiation measurement. The Flight Software (FSW) system is responsible for all mission phases, including launch, cruise, entry-descent-landing, and surface operation of the rover. Because of the essential nature of flight software to project success, each of the software modules is undergoing extensive testing to identify and correct errors.

Whitehill, Robert↗

Three Decades of Wetland Methane Surface Flux Modeling by Earth System Models–Advances, Applications, and Challenges

Earth System Models (ESMs) simulate the exchange of mass and energy between the land surface and the atmosphere, with a key focus on modeling natural greenhouse gas feedbacks. Methane is the second most important greenhouse gas after carbon dioxide. There are growing concerns over the rapidly increasing methane concentration in the atmosphere, underscoring the need for accurate global modeling of its emissions using ESMs. Of the multitude of sources of methane globally, wetlands are the largest natural emitters for methane, leading to significant efforts targeting their representation in ESMs with a special focus on their methane emissions. In this review, we first provide a historical overview of including wetland–methane components in ESMs and how methane modeling approaches have evolved over time. Second, we discuss recent modeling advancements that show promise for improvements in methane emissions predictions, namely the coupling of surface and atmospheric modules of ESMs, the representation of microtopography and transport mechanisms, the resolution of microbial processes at different spatial–temporal scales, and the improved mapping of wetland area extent across the different wetland types. Third, we shed light on the different challenges hindering accurate estimations of wetland–methane emissions, as shown by the consistent discrepancy between bottom–up and top–down models' predictions. Finally, we emphasize that more detailed representation of biogeochemistry and dynamic hydrology while resolving the within–wetland vegetation heterogeneity should improve model predictions, especially when coupled with expanding ground–based measurement networks and high–resolution remote sensing mapping of methane–relevant variables, such as water elevation, water table depth, and methane concentration.

54 ENVIRONMENTAL SCIENCES↗

The role of Ca-bridged organic matter in an alkaline soil, as revealed by multimodal chemical imaging

Mineral–organic matter (OM) studies have predominantly focused on acidic soils that are abundant in iron (Fe) oxides and aluminum (Al) oxides. We have probed mineral–OM interactions in an alkaline or calcareous soil of the Aridisols class. Unlike the role of Fe and Al, the role of Ca-minerals (particularly calcite), which are ubiquitous in alkaline soils, in OM sequestration is not well understood. Multiple recent model studies with aqueous Ca2+ or synthetic calcite and a suite of OM compounds have shown Ca-OM assemblages to be spatially correlated with calcite at the microscale. To study the chemical state of both Ca and Fe and their competing role in soil organic matter (SOM) stabilization, we performed laboratory characterization using x-ray diffraction, Mössbauer spectroscopy, x-ray photoelectron spectroscopy, scanning electron microscopy, and scanning transmission electron microscopy, alongside synchrotron-based microscale chemical imaging using scanning transmission x-ray microscopy combined with near-edge x-ray absorption fine structure. Ca mineral–organic associations were found to be ubiquitous in this system and are likely critical for understanding SOM stabilization/degradation in alkaline soils. From our findings on mineralogy, speciation, and the nature of Ca-OM bridging, we identified differences in C and Ca chemistry based on the relative location of OM to Ca minerals. The OM near the calcite crystal was enriched in lipid and protein moieties, Ca-OM next to Fe minerals displayed a strong contribution from aromatic compounds, while on the surface of microbes, the carbonate was believed to be of microbial in origin, as also suggested by preliminary works reporting on the formation of amorphous calcite or nano-calcite. In Ca-OM admixed with carbonate, it was difficult to distinguish Ca-associated OM from amorphous calcite or nano-calcite.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Depth-dependent Metagenome-Assembled Genomes of Agricultural Soils under Managed Aquifer Recharge

Abstract Managed Aquifer Recharge (MAR) systems, which intentionally replenish groundwater aquifers with excess water, are critical for addressing water scarcity exacerbated by demographic shifts and climate variability. To date, little is known about the functional diversity of the soil microbiome at different soil depth inhabiting agricultural soils used for MAR. Knowing the functional diversity is pivotal in regulating nutrient cycling and maintaining soil health. Metagenomics, particularly Metagenome-Assembled Genomes (MAGs), provide a powerful tool to explore the diversity of uncultivated soil microbes, facilitating in-depth investigations into microbial functions. In a field experiment conducted in a California vineyard, we sequenced soil DNA before and after water application of MAR. Through this process, we assembled 146 medium and 14 high-quality MAGs, uncovering a wide array of archaeal and bacterial taxa across different soil depths. These findings advance our understanding of the microbial ecology and functional diversity of soils used for MAR, contributing to the development of more informed and sustainable land management strategies.

Science & Technology - Other Topics↗

Detection and Screening of Organic Contaminants in A Riverine System of Georgia Using Non-Targeted Analysis

Numerous organic chemicals exist within aquatic environments, yet effectively screening and prioritizing them is a huge challenge. This study provides a comprehensive investigation into the ecological dynamics of the North Oconee River within Athens-Clarke County, Georgia, with a specific focus on the distribution of 33 identified compounds, including a prominent pesticide. The research, conducted in the riverine ecosystems proximal to the Firefly trail, employs advanced analytical techniques to elucidate potential contamination sources arising from agricultural and urban runoff. Intriguingly, the study reveals North Oconee River near the Firefly Trail as a notable site for heightened pesticide contamination, warranting a meticulous exploration of its origins. Furthermore, the investigation unveils the intricate microbial degradation processes of malathion within the North Oconee River, elucidating the pivotal role played by microbial activity in river water. The detection of degradant byproducts prompts the considerations of bioavailability and toxicity, associating potential implications for the river’s overall ecological health. Ongoing research endeavors to precisely quantify environmental risks and unravel indigenous microbial degradation pathways, presenting pivotal contributions to the scientific community’s understanding of complex riverine ecosystems. This research serves as a foundational piece in informing sustainable environmental management practices and emphasizes the urgency of comprehensive stewardship in safeguarding aquatic ecosystems.

54 ENVIRONMENTAL SCIENCES↗

Utilization of on-site resources for Regenerative Life Support Systems at a lunar outpost

Regenerative life support systems (RLSS) will be required to regenerate air, water, and wastes, and to produce food for human consumption during long-duration stays on the moon. It may be possible to supplement some of the materials needed for RLSS from resources on the moon. Natural materials at the lunar surface may be used for a variety of lunar RLSS needs, including (i) soils or solid-support substrates for plant growth, (ii) sources for extraction of essential, plant-growth nutrients, (iii) substrates for microbial populations in the degradation of wastes, (iv) sources of O2 and H, which may be used to manufacture water, (v) feed stock materials for the synthesis of useful minerals (e.g., molecular sieves), and (vi) shielding materials surrounding the outpost structure to protect humans, plants, and microorganisms from harmful radiation.

Ming, D. W.↗

Water Treatment Systems Make a Big Splash

In the 1960s, NASA's Manned Space Center (now known as Johnson Space Center) and the Garrett Corporation, Air Research Division, conducted a research program to develop a small, lightweight water purifier for the Apollo spacecraft that would require minimal power and would not need to be monitored around-the-clock by astronauts in orbit. The 9-ounce purifier, slightly larger than a cigarette pack and completely chlorine-free, dispensed silver ions into the spacecraft s water supply to successfully kill off bacteria. A NASA Technical Brief released around the time of the research reported that the silver ions did not impart an unpleasant taste to the water. NASA s ingenuity to control microbial contamination in space caught on quickly, opening the doors for safer methods of controlling water pollutants on Earth.

Source record↗

Microbial Communities in Microgravity: Simulation in Lab and on the Computer

Microorganisms grow differently in spaceflight than they do on Earth. While much remains unexplained about how microgravity affects microbial growth, one dominant hypothesis is that the lack of density-driven convection in the liquid growth environment makes mixing diffusion-limited, and therefore slower. This is supported by evidence that individual microbial strains experience starvation and acid stress in microgravity. However, if it is true, then microgravity would also have measurable effects on microbes in mixed communities, because many interspecies interactions involve the exchange of soluble metabolites through the medium (cross-feeding). Specifically, cooperative cross-feeding communities would grow more slowly in microgravity, and cooperation would be less stable on evolutionary timescales. Here we describe our efforts to test this hypothesis by simulating microgravity in silico and in the lab, using a model system of Eschericia coli and Salmonella enterica that grow only when they can exchange methionine and acetate. We created CAMDLES (CFD-DEM Artificial Microgravity Developments for Living Ecosystem Simulation) as an extension of CFDEM®coupling software, to carry out computational modeling of biological flows, growth, and mass transfer in microgravity and also in laboratory artificial microgravity devices (rotating wall vessels, RWV). Using CAMDLES, we found distinct differences in growth rates between RWV and true microgravity, and we were able to identify several features, such as spatial distribution, biofilm formation, and product yield parameters, that influence the degree to which RWV growth recapitulates microgravity growth. In addition, we report on the development of a laboratory system for monitoring growth rates and species ratios of the community in RWVs, using fluorescent strains. Pairing CAMDLES with the laboratory model system allows us to generate quantitative predictions about the effects of spaceflight on organisms that will be essential to sustaining human space exploration in the long term.

microbiology↗

Real-time monitoring and prediction of water quality parameters and algae concentrations using microbial potentiometric sensor signals and machine learning tools

We report the overarching hypothesis of this study was that temporal microbial potentiometric sensor (MPS) signal patterns could be used to predict changes in commonly monitored water quality parameters by using artificial intelligence/machine learning tools. To test this hypothesis, the study first examines a proof of concept by correlating between MPS's signals and high algae concentrations in an algal cultivation pond. Then, the study expanded upon these findings and examined if multiple water quality parameters could be predicted in real surface waters, like irrigation canals. Signals generated between the MPS sensors and other water quality sensors maintained by an Arizona utility company, including algae and chlorophyll, were collected in real time at time intervals of 30 min over a period of 9 months. Data from the MPS system and data collected by the utility company were used to train the ML/AI algorithms and compare the predicted with actual water quality parameters and algae concentrations. Based on the composite signal obtained from the MPS, the ML/AI was used to predict the canal surface water's turbidity, conductivity, chlorophyll, and blue-green algae (BGA), dissolved oxygen (DO), and pH, and predicted values were compared to the measured values. Initial testing in the algal cultivation pond revealed a strong linear correlation (R 2 = 0.87) between mixed liquor suspended solids (MLSS) and the MPSs' composite signals. The Normalized Root Mean Square Error (NRMSE) between the predicted values and measured values were <6.5%, except for the DO, which was 10.45%. The results demonstrate the usefulness of MPSs to predict key surface water quality parameters through a single composite signal, when the ML/AI tools are used conjunctively to disaggregate these signal components. The maintenance-free MPS offers a novel and cost-effective approach to monitor numerous water quality parameters at once with relatively high accuracy.

54 ENVIRONMENTAL SCIENCES↗

Comparing first- and second-generation bioethanol by-products from sugarcane: Impact on soil carbon and nitrogen dynamics

The first- and second-generation bioethanol by-products (bagasse and fermentation by-product, respectively) have different biochemical characteristics relating to carbon (C) and nitrogen (N) due to their production processes. To examine the impacts of fermentation by-product and bagasse on soil C and N dynamics, a 120 day laboratory incubation experiment was conducted by applying these amendments in soil at an equal carbon application rate (2.2 g C kg -1 soil). There was a significant increase in overall cumulative CO 2 -C production from amended soil compared to the control soil, though the loss was greatest in bagasse amended soil. However, δ13C-CO 2 measurements suggested that fermentation by-product addition suppressed native soil C mineralization and prompted strong negative C priming (-57%). In contrast, bagasse amendment maintained similar native soil C mineralization to control soil. Decreased microbial biomass turnover time appears to be the major driving force for increased CO 2 -C production and soil C loss following bioethanol by-product amendments. Increased mineral N production indicated net N mineralization after fermentation by-product addition, while addition of bagasse resulted in lower N availability and N immobilization. Overall, results suggested that fermentation by-product application is advantageous in maintaining native soil C stock and N mineralization compared to bagasse. Thus, in a sustainable second-generation bioethanol production system, bagasse may be used for second-generation bioethanol production and the cogenerated by-product could be used as an amendment to improve soil C stock and N availability.

09 BIOMASS FUELS↗

Tracing priming effects in palsa peat carbon dynamics using a stable isotope-assisted metabolomics approach

Introduction: Peatlands store up to a third of global soil carbon, and in high latitudes their litter inputs are increasing and changing in composition under climate change. Although litter significantly influences peatland carbon and nutrient dynamics by changing the overall lability of peatland organic matter, the physicochemical mechanisms of this impact—and thus its full scope—remain poorly understood. Methods: We applied multimodal metabolomics (UPLC-HRMS, 1 H NMR) paired with 13 C Stable Isotope-Assisted Metabolomics (SIAM) to track litter carbon and its potential priming effects on both existing soil organic matter and carbon gas emissions. Through this approach, we achieved molecule-specific tracking of carbon transformations at unprecedented detail. Results: Our analysis revealed several key findings about carbon dynamics in palsa peat. Microbes responded rapidly to litter addition, producing a short-term increase in CO 2 emissions, fueled nearly exclusively by transformations of litter carbon. Litter inputs significantly contributed to the organic nitrogen pool through amino acids and peptide derivatives, which served as readily accessible nutrient sources for microbial communities. We traced the fate of plant-derived polyphenols including flavonoids like rutin, finding evidence of their degradation through heterocyclic C-ring fission, while accumulation of some polyphenols suggested their role in limiting overall decomposition. The SIAM approach detected subtle molecular changes indicating minimal and transient priming activity that was undetectable through conventional gas measurements alone. This transient response was characterized by brief microbial stimulation followed by rapid return to baseline metabolism. Pre-existing peat organic matter remained relatively stable; significant priming of its consumption was not observed, nor was its structural alteration. Discussion: This suggests that while litter inputs temporarily increase CO 2 emissions, they don’t sustain long-term acceleration of stored carbon decomposition or substantially decrease peat’s carbon store capacity. Our findings demonstrate how technological advancements in analytical tools can provide a more detailed view of carbon cycling processes in complex soil systems.

54 ENVIRONMENTAL SCIENCES↗