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Plant-specific microbial diversity facilitates functional redundancy at the soil-root interface

Abstract Aims Plant-specific microbial diversity reflecting host-microbe coevolution was frequently shown at the structural level but less on the functional scale. We studied the microbiome of three compartments at the soil root interface (root endosphere, rhizosphere, bulk soil) of medicinal plants cultivated under organic management in Egypt. The study aimed to examine the impact of the rhizosphere on microbial community composition and diversity in desert agricultural soil, as well as to identify specific functions associated with the rhizosphere. Methods The microbiome community structure, diversity, and microbial functioning were evaluated through the utilization of 16S rRNA gene amplicon and shotgun metagenome sequencing. Results We found the typical rhizosphere effect and plant-species-specific enrichment of bacterial diversity. The annual plants Calendula officinalis and Matricaria chamomilla ( Asteraceae ) were more similar than the perennial Solanum distichum ( Solanaceae ). Altogether, plant species explained 50.5% of the variation in bacterial community structures in the rhizosphere. Our results indicate a stronger effect of the plant species in terms of modulating bacterial community structures in the rhizosphere than in root endosphere samples. The plant-driven rhizosphere effect could be linked to redundant plant beneficial functions in the microbiome, while enrichment of specific genes related to amino acid ion transport and metabolism, carbohydrate transport and metabolism, defense mechanisms, and secondary metabolites biosynthesis were more specific. Conclusions The study explores the microbiome continuum at the soil-root interface of medicinal plant species, revealing significant bacterial community structure shifts and plant specificity. The study provides insights into the essential microbiome components contributing to rhizosphere functionality.

Wicaksono, Wisnu Adi (ORCID:0000000215561981)

Spatial and Seasonal Patterns of the Mosquito Community in Central Oklahoma

Mosquitoes (Culicidae) are ubiquitous flying insects that function as vectors for several viruses that cause disease in humans. Mosquito abundance and diversity are influenced by landscape features and environmental factors such as temperature and precipitation and vary across seasons and years. The range and phenology of many mosquito species that vector viruses relevant to human health are changing. We sampled mosquito communities in central Oklahoma for four years at thirteen sites, collecting over 25,000 mosquitoes; among these, we identified 27 different species, including several that transmit human pathogens and were collected in suburban backyards. Community composition differed across the landscape and changed from early season to late season and year to year. This effort to describe mosquito communities in Oklahoma is a first step toward assessing and predicting arbovirus risk, an ongoing and dynamic public health challenge.

David Hoekman

Bacterial and fungal composition and exometabolites control the development and persistence of soil water repellency

Soil water repellency (SWR), the reduced affinity of soil for water, is a phenomenon that affects soils globally. With worsening climate change, SWR is expected to increase emphasizing the need to understand the mechanisms driving SWR development and persistence. The importance of the soil microbes in SWR has been postulated for decades, but limited research has been conducted into whole-community interactions and the role of community metabolic activity. To address this gap in knowledge, we investigated the direct effect of microbial community composition, activity, and diversity, as well as their associated metabolites on the development and persistence of SWR by inoculating microcosms containing model soils with 15 different microbial communities and quantified respiration and SWR over time. Six communities that consistently produced either a hydrophobic or hydrophilic phenotype were characterized using metagenomics and metabolomics to determine the impact of microbial and metabolite composition and diversity on SWR. We identified several bacterial genera with significant changes in abundance between SWR phenotypes including Nocardiopsis and Kocuria in hydrophilic and Streptomyces and Cutibacterium in hydrophobic. We discovered that hydrophilic communities were more positively connected when compared to hydrophobic communities, which could be due to an increase in defense mechanism genes. Additionally, we identified specific metabolites associated with hydrophilic and hydrophobic phenotypes including an increase in the osmolyte ectoine in hydrophilic and an increase in plant-derived decomposition products in hydrophobic communities. Finally, our research suggests that fungi, previously thought to cause hydrophobicity, may actually contribute to hydrophilicity through their preferential consumption of hydrophobic compounds.

54 ENVIRONMENTAL SCIENCES

Retrospectives: Current State of Knowledge on the Intersection of Spaceflight Stressors and Microbial Risks to Crew and Craft

OVERVIEW The spaceflight environment has several unique stressors that affect the health of both the crew and the spacecraft. An area of continued, albeit incomplete, study is the interaction of these stressors on microbial populations inherent to both astronauts and spacecraft surfaces and systems. A primary concern is the potential for the spaceflight environment to perturb the phenotype of these populations towards negative outcomes for crew and craft. In order to effectively mitigate these potential risks, they must first be characterized. We performed a retrospective literature analysis to assess the current state of knowledge regarding the affects of ionizing radiation and elevated CO2 on relevant microbial populations. The results of these retrospectives will guide next steps in the decisions of what (if any) further studies should be pursued and to guide decisions of the need for countermeasures. STRESSORS Ionizing radiation. The health risk involved with increased exposure to cosmic radiation has been studied in crew for 35+ years, with human health and cancer risk being the main focus. However, space radiation could also affect both the resident microorganisms aboard the ISS and the normal, healthy astronaut microbiomes that are of direct concern for crew health. A retrospective review of over 250 publications was accomplished looking at the impact of cumulative ionizing radiation doses lower than 3 Gy (chronic or acute) on microbial populations. Elevated CO2. The health risk involved with elevated atmospheric CO2 in spacecraft, primarily focusing on human toxicological risks, is understudied. The current Spaceflight Maximum Allowance Concentration for 24-hour average CO2 is 0.4% (3 mm Hg), which is significantly higher than terrestrial levels (0.04%). Whether these elevated ambient CO2 levels aboard spacecraft influence the diversity and phenotypic responses of the resident microbial communities from both the spacecraft environment (air, surface, water) and crew members (gut, nasal, skin microbiomes) is not known. A retrospective review was accomplished looking at the impact of chronic CO2 exposure up to 0.7% (5 mm Hg) for up to 6 months and acute exposure up to 2.6% (20 mm Hg) for up to 24 hours. CONCLUSIONS: MICROBIOME OF THE BUILT ENVIRONMENT The microbiome of the built spacecraft environment has been sampled consistently over the course of human spaceflight and significant advancements have been made in identifying microbial populations on the ISS. The dominant source of microbes on spacecraft surfaces are human-derived. Once in the spacecraft built environment, the extreme environment selects for features that enhance survival. While efforts to understand potential antibiotic resistance and pathogenicity of ISS isolates is robust, there is little to no understanding of which spaceflight environmental stressors, to include ionizing radiation or elevated CO2, drive the evolutionary trajectory of spacecraft-associated microbial populations. CONCLUSIONS: MICROBE-HOST INTERACTIONS The host-microbiome field has emerged as an important factor in human health on Earth as well in spaceflight. The field is struggling with the complexity of the system under investigation as there is substantial taxonomic and functional heterogeneity in these communities, making it difficult to establish clear stimulus-response dynamics. Taxonomic characterization is the norm; however, the functional role of each community member is key to linking environmental perturbations to potential dysbiosis. For both ionizing radiation and elevated CO2, the likely target of the perturbation is the host tissue, not the microbes themselves.. Any resulting changes to the microbial community composition and/or function is likely a result of adapting to those changes in the host physiology. RECOMMENDATIONS Emphasize functional characterization as opposed to taxonomic characterization of microbial communities. Increase the number of investigations using chronic, spaceflight-relevant doses of ionizing radiation. MoBE studies should move away from observational studies towards predictive modeling of community dynamics. Continue to develop scale-down models, such as tissues-on-a-chip & defined microbial communities. Focus on the crew response to elevated CO2 over MoBE considerations. Assess how direct contact with the hypercapnic environment affects skin microbiome dynamics.

retrospective

Meta Biome: a multiscale model integrating agent-based and metabolic networks to reveal spatial regulation in gut mucosal microbial communities

ABSTRACT Mucosal microbial communities (MMCs) are complex ecosystems near the mucosal layers of the gut essential for maintaining health and modulating disease states. Despite advances in high-throughput omics technologies, current methodologies struggle to capture the dynamic metabolic interactions and spatiotemporal variations within MMCs. In this work, we presentMetaBiome, a multiscale model integrating agent-based modeling (ABM), finite volume methods, and constraint-based models to explore the metabolic interactions within these communities. Integrating ABM allows for the detailed representation of individual microbial agents each governed by rules that dictate cell growth, division, and interactions with their surroundings. Through a layered approach—encompassing microenvironmental conditions, agent information, and metabolic pathways—we simulated different communities to showcase the potential of the model. Using ourin-silicoplatform, we explored the dynamics and spatiotemporal patterns of MMCs in the proximal small intestine and the cecum, simulating the physiological conditions of the two gut regions. Our findings revealed how specific microbes adapt their metabolic processes based on substrate availability and local environmental conditions, shedding light on spatial metabolite regulation and informing targeted therapies for localized gut diseases.MetaBiome provides a detailed representation of microbial agents and their interactions, surpassing the limitations of traditional grid-based systems. This work marks a significant advancement in microbial ecology, as it offers new insights into predicting and analyzing microbial communities. IMPORTANCE Our study presents a novel multiscale model that combines agent-based modeling, finite volume methods, and genome-scale metabolic models to simulate the complex dynamics of mucosal microbial communities in the gut. This integrated approach allows us to capture spatial and temporal variations in microbial interactions and metabolism that are difficult to study experimentally. Key findings from our model include the following: (i) prediction of metabolic cross-feeding and spatial organization in multi-species communities, (ii) insights into how oxygen gradients and nutrient availability shape community composition in different gut regions, and (iii) identification of spatiallyregulated metabolic pathways and enzymes inE. coli. We believe this work represents a significant advance in computational modeling of microbial communities and provides new insights into the spatial regulation of gut microbiome metabolism. The multiscale modeling approach we have developed could be broadly applicable for studying other complex microbial ecosystems.

Microbiology

Cyanobacterial Community Structure In Lithifying Mats of A Yellowstone Hotspring-Implications for Precambrian Stromatolite Biocomplexity

Denaturing Gradient Gel Electrophoresis (DGGE) of partial 16S rRNA gene sequences was used to investigate the molecular biodiversity of cyanobacterial communities inhabiting various lithified morpho-structures in two hotsprings of Yellowstone National Park. These morpho-structures - flat-topped columns, columnar cones, and ridged cones - resemble ancient stromatolites, which are possibly biogenic in origin. The top, middle and bottom sections of these lithified morpho-structures, as well as surrounding non-lithified mats were analyzed to determine the vertical and spatial distribution of cyanobacterial communities. Results from DGGE indicate that the cyanobacterial community composition of lithified morpho-structures (flat-topped columns, columnar cones, and ridged cones) were largely similar in vertical distribution as well as among the morpho-structures being studied. Preliminary results indicate that the cyanobacterial communities in these lithified morpho-structures were significantly different from communities in surrounding non-lithified mats. These results provide additional support to the theory that certain Phormidium/Leptolyngbya species are involved in the morphogenesis of lithifying morpho-structures in hotsprings and may have played a role in the formation of ancient stromatolites.

Lau, Evan

Contrasting effects of land-use and local disturbance on plant and pollinator communities in wetlands

While pollinators and wetlands both provide important ecosystem services (e.g., the pollination of flowering plants and improving water quality), the relationship between the two is not well understood. Both biotic and abiotic effects can mediate the local wetland flower and pollinator community. In this study, we investigated how land use, including a land use gradient at five different radii, from 250 m to 2 km, along with anthropogenic disturbance affected pollinators in wetland ecosystems. We surveyed the abundance and diversity of plant-pollinator communities in fifteen different wetlands across two years. We also tested the relationship between water quality and temperature, and the abundance and diversity of flowering plants and pollinating insects. Our results suggest that increasing temperature, which was strongly associated with developed land use, had a negative effect on the floral display of wetland plants, as well as the abundance of all flower visitors and hover flies. Hover fly abundance was also positively associated with agricultural land use and total nitrogen in the water. Meanwhile, the abundance of female bees was affected by an interaction between temperature and disturbance: female bees were most abundant when temperatures were lower in areas of low disturbance. In contrast, pollinator species richness increased with temperature when developed land use was low, and floral diversity was strongly affected by several interactions between disturbance, land use, and water quality. Finally, the community composition of both plants and insects varied significantly among low, medium, and high disturbance categories, with weedier, non-native species being significantly associated with areas of higher disturbance and in sites with greater anthropogenic land use. We demonstrate that ecological communities shift significantly in response to anthropogenic change. Our work also illustrates the importance of quantifying interactions between land use and local disturbance with abiotic factors such as temperature and water quality on ecological systems.

Disturbance

Observing Ocean Ecosystem Responses to Volcanic Ash

Volcanic eruptions can be catastrophic events, particularly when they occur in inhabited coastal environments. They also play important roles in climate and biogeochemical cycles, including through nutrient deposition in the ocean. Volcanic ash studies in the ocean have focused on the phytoplankton response, generally quantifying changes in chlorophyll-a concentration. Many gaps remain in addressing fundamental questions regarding why volcanic ash deposition may enhance or limit both phytoplankton growth and/or drive community composition shifts. Here we outline a wide, multidisciplinary vision for monitoring volcanic eruptions near ocean ecosystems from satellites, including considerations for characteristics of airborne volcanic ash and ash geochemistry in seawater. Ultimately, observations beyond chlorophyll-a are needed to quantify phytoplankton communities (including harmful algal blooms) and possible impacts across higher trophic levels. We synthesize relevant research from volcanic studies as well as atmospheric and ocean sciences to identify the ‘known unknowns’ in ash-ecosystem studies. Our goal is to move toward an improved understanding of how real-time and near-real-time monitoring of volcanic eruptions can help address societally relevant questions.

volcano

Microbial ecology of acidic, biogenic gypsum: community structure and distribution of extremophiles on freshly formed and relict sulfate deposits in a hydrogen sulfide-rich cave

Sulfate minerals are abundant on the Martian surface, and many of these evaporite deposits are thought to have precipitated from acidic fluids. On Earth, gypsum (CaSO 4 •2H 2 O) and other sulfates sometimes form under acidic conditions, so exploring the extremophilic life that occurs in these mineral environments can help evaluate the astrobiological potential of acid sulfate depositional settings. Here, we characterized the microbial communities associated with acidic gypsum deposits in a sulfuric acid cave, where sulfate precipitation is driven by sulfide-oxidizing bacteria and archaea. We used 16S rRNA gene sequencing and cell counts to characterize gypsum-associated microorganisms in freshly formed and relict deposits throughout the cave, to test how microbial community composition and abundance would vary with distance from the sulfidic water table and with the concentration of H 2 S(g) and other gases in the cave atmosphere. We found that actively forming gypsum in the lower cave levels was colonized by low-diversity communities that have few cells compared to other environments in the cave. The most abundant taxa were Acidithiobacillus, Metallibacterium, Mycobacteria, and three different Thermoplasmatales-group archaea, which occupied distinct niches based on proximity to sulfidic streams and the concentration of gases in the cave air. By contrast, deposits in older cave levels had more diverse communities that were distinct from those associated with freshly formed gypsum and likely represent a community reliant on different energy resources. These findings show that acidic sulfate deposits serve as habitats for extremophilic microorganisms and broaden our knowledge of the life associated with terrestrial sulfates.

58 GEOSCIENCES

Geological and vegetational applications of Shuttle Imaging Radar-B, Mineral County, Nevada

Multiple-incidence angle and multi-azimuth radar data were acquired from a Shuttle platform over test sites in Nevada in October 1984. An attempt was made to correlate these data with ground features for the purpose of evaluating the use of such data for geological and vegetational assessment. Standard ecological parameters with respect to the flora (community composition, dominance, and relative cover) were recorded in the field at the time of overflight. Although a total of 33 species representing 11 plant families were recognized, and plant cover ranged from 13 to 26 percent, radar data could not be used to separate plant communities. The signal return is more a function of abiotic conditions than vegetative characteristics. Illumination geometry plays an important role in the ability to detect strike-slip and dip-slip faults. Local incidence angle is the most important parameter, and SIR-B data takes with small incidence angles are superior for identifying certain styles of faulting. Look direction is critical for detecting faults with a dip-slip component. New structural features were not observed. Problems with radar antenna power and recording significantly affected data quality.

Borengasser, M. X.

Hyporheic zone, river, and groundwater metagenome resolved genomes and rpS3 genes in East River Watershed, Colorado USA Summer 2020, 2021

Here we present metagenome assembled genomes (MAGs) for the bacterial and archaeal communities from water filter collected across 8 locations along the East River Watershed, CO, and 1 nearby groundwater well. The purpose was to look for connectivity and similarities across the network and to see the impact of the groundwater. As a part of Lawrence Berkeley National Laboratory (LBNL) Watershed Science Focus Area (SFA), we assessed community composition and strain similarities between the sites and we also compared it to previous metagenomic studies within the watershed looking at floodplain (Matheus Carnevali et al. 2021) and hillslope (Lavy et al. 2019) microbiomes. Here we present metagenome assembled genomes (MAGs) for the bacterial and archaeal communities from filters across 8 locations during August 2020 and July 2021. This resulted in 32 samples. The groundwater sample was sequenced at UC Berkley's QB3. The other 31 samples were sequenced at University of Maryland. Metagenomes were assembled using four autobinners and the best bins were selected using dasTool. The genomes were dereplicated at 95% with dRep and the subset of winning genomes were manually curated based on visual inspection of taxonomic profile, GC content, coverage, and a set of 51 bacterial single copy genes (BSCG), and 38 archaeal signal copy genes (ASCG). The dataset includes a zip file of 311 genomes (HZ_River_SW_MAGS_Dereplicated_95.zip). The dataset additionally includes a zipped file of ribosomal protein small subunit 3 (rpS3) proteins from the hyporheic zone and river data (rpS3_Proteins_HZ_River.zip), a metadata file used to register associated samples with IGSNs (International Generic Sample Numbers) (samples.csv), a location metadata file (locations.csv). This work was supported by the Watershed Function Science Focus Area at Lawrence Berkeley National Laboratory funded by the US Department of Energy, Office of Science, Biological and Environmental Research under Contract No. DE-AC02-05CH11231.

DNA

Seeding Advanced Treated Wastewater for Purposes of Direct Potable Reuse

Direct potable reuse (DPR) is a promising solution to address water scarcity. However, a better understanding of how introducing advanced treated water (ATW) affects microbial communities present in distribution systems is needed. Here, in this study, we measured changes to the microbial water quality in simulated distribution systems that were conditioned using treated, unimpaired surface water (SW) and then transitioned to ATW. In addition, we investigated whether adding a biological filtration step would seed the microbial community of the ATW and whether the influence would persist in the simulated distribution systems. We found that the bulk water in the ATW-fed distribution systems had lower cell counts and ATP concentrations and a distinct microbial community (based on 16S amplicon sequencing) compared to the SW-fed or the seeded ATW-fed systems. However, biofilm community composition and biomass remained consistent regardless of the feedwater. Increased microbial biomass and diversity were present in the seeded ATW, with several amplicon sequence variants identified as being introduced by the biological filter. Our results suggest that directly introducing ATW to distribution systems could disturb the existing microbial community. Preparing ATW for distribution via biological filtration may deliver more predictable and stable microbial water quality than introducing unseeded ATW.

16S

Belowground plant carbon and nitrogen exchange: plant-derived carbon inputs and pore structure formation

Belowground plant transfer of carbon (C) and nitrogen (N) can benefit soil ecosystems, increasing soil C gains and plant N availability, while improving soil pore structure. We explored such transfers among three plant species of North American prairie, where C and N were transferred from a grass (Panicum virgatum L., switchgrass (Sgrass)) to either a legume (Lespedeza capitata Michx., bush clover (Bclover)), a forb (Rudbeckia hirta L., black-eyed Susan (BSforb)), or a mixture of the two. The plants were grown either with/out direct root contact, thus allowing assessment of the relative contributions of fungal- and root-based transfer pathways. The Sgrass was labeled with 13 C and 15 N, and C and N transfers were assessed by measuring isotope enrichment of roots and aboveground biomass of neighboring plants. Soil inputs of plant-derived C and N were assessed by isotope analyses of the rhizosphere soil. X-ray computed tomography was used for pore structure analyses. Carbon transfer was much higher in the presence of direct/close root contact between source and recipient plants, yet N transfers appeared to be mainly fungal driven. Here, while C and N were readily transferred from Sgrass to other Sgrass and Bclover neighbors, transfers to BSforb were negligible. However, in a three species system, the presence of the legume enhanced C and N transfers to BSforb, suggesting non-additive influences of diverse plant community composition. The more plant-derived C and N was found in the rhizosphere of recipient plants, the greater C and N transfers through roots. Greater C and N transfers were associated with increases in 8–30 μm diameter pores and decreases in >150 μm pores. Summarily, diverse plant communities, especially those with legumes, increase C and N transfers, which then benefit soil C inputs and its protection via changes in pore structure.

13 CO 2 pulse labeling

Evolutionary Stability of Microbial Mutualism in Simulated Microgravity

Sustainable long-term space travel will require astronauts to live in built environments cohabited by microbes. Though much of space biology research has focused on microbial monocultures, microbes naturally live in multispecies communities in which they compete and cooperate. Understanding how communities experience spaceflight over generations can influence sanitation measures and the design of synthetic communities for applications in spaceflight. In this study, we investigated the evolutionary stability of mutualism in multispecies communities in microgravity using a model system of cooperating microbes. Our model consisted of bioengineered strains of mutually dependent E. coli and S. enterica. E. coli metabolized lactose and produced a carbon source for S. enterica, which secreted methionine for E. coli. By introducing a noncooperative ("cheater") strain of S. enterica, we tested the dynamic stability of mutualism. Using a rotating wall vessel, we simulated microgravity, specifically a low-shear, diffusion-limited environment where metabolite transfer is limited compared to 1g conditions. Fluorescent motile and non-motile strains were used to assess how motility affects the microbial experience of microgravity. We hypothesized that mutualism in non-motile cocultures would be favored in microgravity due to the diffusion-limited spatial structure, while motile strains would show less difference between simulated microgravity and other conditions. We cultured communities for approximately 20 generations and assessed community composition over time using flow cytometry. We compared simulated microgravity to cultures grown in agar plates as spatially structured controls, shaken flasks as well-mixed controls, and still flasks representing the convective mixing of 1g. A high, sustained cooperator-to-cheater ratio represented stable mutualism within the community. Our results show that microbial communities may better maintain mutualism in microgravity compared to their counterparts in 1g conditions and that the motility of a community’s constituents influences how the community experiences microgravity on generational timescales.

Nathan Moreno

Enhancing climate-smart crop performance in arid agrivoltaics systems: effects of photovoltaic shading and soil amendments on tepary bean growth, yield, and associated soil microbiome

As climate change expands the world’s arid and semiarid regions, sustainable systems that integrate food and energy production are becoming increasingly critical. Agrivoltaics—co-locating crops with photovoltaic (PV) panels—offers a dual land-use strategy that mitigates environmental stress by shading crops, conserving soil moisture, and enhancing PV efficiency. While climate-smart crops like the tepary bean ( Phaseolus acutifolius ) are well adapted to heat and drought, little is known about how these crops and their associated soil microbiomes respond to the unique microclimates created by PV shading. This study evaluated tepary bean performance and plant–microbial interactions under PV-shade vs. no shade across three soil amendment treatments at two experimental sites. We assessed plant traits including germination, phenology, biomass, height, as well as yield and bean morphology, alongside shifts in soil microbial composition and functional potential. Plants grown under PV-shade were generally taller, with extended reproductive periods and higher yields: 42% of shaded plants produced beans compared to only 8% under full sun. Shaded plants also produced rounder, higher-quality beans, whereas non-shaded plants yielded flatter, less developed beans. Microbial community composition was more strongly influenced by amendment and site conditions than by shading alone. Key microbial taxa (e.g., Glomeromycetes, Desulfobacterota ) and predicted functions (e.g., denitrification, nitrogen-respiration, sulfate reduction) were associated with differences in plant performance. Finally, combining agrivoltaic systems with targeted soil amendments can enhance crop yield and soil microbial functionality—offering a promising strategy for sustainable agriculture in arid landscapes.

14 SOLAR ENERGY

Response of soil nutrient pools and microbiomes to recurrent wildfire disturbance and varying burn severities in a mixed conifer forest

Wildfire is a pervasive disturbance in mixed-conifer forests, yet the relative influence of fire recurrence versus burn severity on soil biogeochemistry and microbial communities remains poorly quantified. We examined a natural gradient of fire history (0–3 prior fires) and burn severity (low–high) spanning 50 yr in a mixed-conifer ecosystem to assess how repeated fire shapes soil carbon (C) and nitrogen (N) pools, their isotopic signatures, mineral and particulate fractions, microbial community composition, carbon-use, CO₂ fluxes, and vegetation cover. Successive fires produced progressively higher bare-ground percentages and lower tree cover, which were tightly linked to declines in microbial diversity and reductions bulk %C, and %N. δ 13 C increased with fire frequency, indicating preferential loss of labile C through combustion or enhanced microbial oxidation, thereby explaining the observed net soil-C decline. Conversely, δ 15 N decreased and pH increased as tree density declined, reflecting altered N cycling and reduced acidification in post-fire soils. Fire recurrence, more than severity, corresponded with a marked shift in the bacterial community: for example, Xanthobacteraceae—key N-fixers and C-cyclers—diminished, while N-fixing Bacillaceae increased, underscoring the tightly coupled nature of soil nutrient dynamics and microbiome composition after repeated burns. Our results demonstrate that fire recurrence appears to be a stronger driver of post-fire soil ecosystem responses in this mixed-conifer forest, influencing both abiotic nutrient pools and the functional potential of the soil microbiome. These findings provide a more enhanced assessment and understanding to date of the biogeochemical consequences of repeated wildfire disturbance that can be used to inform management strategies aimed at preserving soil health in fire-prone landscapes.

54 ENVIRONMENTAL SCIENCES

Retrospectives: Intersection of Spaceflight Stressors and Microbial Risk to Crew and Craft

OVERVIEW The spaceflight environment has several unique stressors that affect the health of both the crew and the spacecraft. An area of continued, albeit incomplete, study is the interaction of these stressors on microbial populations inherent to both astronauts and spacecraft surfaces and systems. A primary concern is the potential for the spaceflight environment to perturb the phenotype of these populations towards negative outcomes for crew and craft. In order to effectively mitigate these potential risks, they must first be characterized. We performed a retrospective literature analysis to assess the current state of knowledge regarding the affects of ionizing radiation and elevated CO2 on relevant microbial populations. The results of these retrospectives will guide next steps in the decisions of what (if any) further studies should be pursued and to guide decisions of the need for countermeasures. STRESSORS Ionizing radiation. The health risk involved with increased exposure to cosmic radiation has been studied in crew for 35+ years, with human health and cancer risk being the main focus. However, space radiation could also affect both the resident microorganisms aboard the ISS and the normal, healthy astronaut microbiomes that are of direct concern for crew health. A retrospective review of over 250 publications was accomplished looking at the impact of cumulative ionizing radiation doses lower than 3 Gy (chronic or acute) on microbial populations. Elevated CO2. The health risk involved with elevated atmospheric CO2 in spacecraft, primarily focusing on human toxicological risks, is understudied. The current Spaceflight Maximum Allowance Concentration for 24-hour average CO2 is 0.4% (3 mm Hg), which is significantly higher than terrestrial levels (0.04%). Whether these elevated ambient CO2 levels aboard spacecraft influence the diversity and phenotypic responses of the resident microbial communities from both the spacecraft environment (air, surface, water) and crew members (gut, nasal,skin microbiomes) is not known. A retrospective review was accomplished looking at the impact of chronic CO2 exposure up to 0.7% (5 mm Hg) for up to 6 months and acute exposure up to 2.6% (20 mm Hg) for up to 24 hours. CONCLUSIONS: MICROBIOME OF THE BUILT ENVIRONMENT The microbiome of the built spacecraft environment has been sampled consistently over the course of human spaceflight and significant advancements have been made in identifying microbial populations on the ISS. The dominant source of microbes on spacecraft surfaces are human-derived. Once in the spacecraft built environment,the extreme environment selects for features that enhance survival. While efforts to understand potential antibiotic resistance and pathogenicity of ISS isolates is robust, there is little to no understanding of which spaceflight environmental stressors, to include ionizing radiation or elevated CO2, drive the evolutionary trajectory of spacecraft-associated microbial populations. CONCLUSIONS: MICROBE-HOST INTERACTIONS The host-microbiome field has emerged as an important factor in human health on Earth as well in spaceflight. The field is struggling with the complexity of the system under investigation as there is substantial taxonomic and functional heterogeneity in these communities, making it difficult to establish clear stimulus-response dynamics.Taxonomic characterization is the norm; however, the functional role of each community member is key to linking environmental perturbations to potential dysbiosis. For both ionizing radiation and elevated CO2, the likely target of the perturbation is the host tissue, not the microbes themselves.. Any resulting changes to the microbial community composition and/or function is likely a result of adapting to those changes in the host physiology. RECOMMENDATIONS Emphasize functional characterization as opposed to taxonomic characterization of microbial communities.Increase the number of investigations using chronic, spaceflight-relevant doses of ionizing radiation. MoBE studies should move away from observational studies towards predictive modeling of community dynamics. Continue to develop scale-down models, such as tissues-on-a-chip & defined microbial communities. Focus on the crew response to elevated CO2 over MoBE considerations. Assess how direct contact with the hypercapnic environment affects skin microbiome dynamics.

Countermeasures

Composition, Growth, Succession, and Function in the Cladophora Microbiome: Insights From Quantitative Stable Isotope Probing and NanoSIMS Imaging

The branching green macroalga Cladophora glomerata and its epiphytic microbiome dominate summer biomass in the Eel River, a Northern California river under Mediterranean (summer drought, winter rain) seasonality. Green Cladophora streamers proliferate in early summer, then change to yellow and then red-brown as epiphyte loads increase. Here, we characterised successional changes in epiphytic bacteria on Cladophora, examining both community composition and growth rates, using quantitative Stable Isotope Probing (qSIP) and 16S rRNA gene amplicon sequencing. The number of bacterial taxa increased with succession while growth rates peaked in the middle stage. NanoSIMS imaging confirmed high sulphur (S) concentrations in Cladophora cell walls relative to surrounding biomass, coinciding with a bloom of sulphur bacteria (bacteria that reduce or oxidise sulphur/sulphates). In general, relative abundances and growth rates were independent, indicating that either metric alone is insufficient for understanding how taxonomy and functional groups affect ecosystem processes. For instance, the relative abundance of nitrogen fixers peaked in the late summer when their relative growth rates were slowest. Such patterns may be driven by space competition limiting growth. Together, changes in abundance and relative growth rates suggest different limiting factors for different functional groups in the Cladophora microbiome at multiple successional stages.

Biological and medical sciences