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At least 55 records · Page 3

From wolves to humans: oral microbiome resistance to transfer across mammalian hosts

The mammalian mouth is colonized by complex microbial communities, adapted to specific niches, and in homeostasis with the host. Individual microbes interact metabolically and rely primarily on nutrients provided by the host, with which they have potentially co-evolved along the mammalian lineages. The oral environment is similar across mammals, but the diversity, specificity, and evolution of community structure in related or interacting mammals are little understood. Here, we compared the oral microbiomes of dogs with those of wild wolves and humans. In dogs, we found an increased microbial diversity relative to wolves, possibly related to the transition to omnivorous nutrition following domestication. This includes a larger diversity of Patescibacteria than previously reported in any other oral microbiota. The oral microbes are most distinct at bacterial species or strain levels, with few if any shared between humans and canids, while the close evolutionary relationship between wolves and dogs is reflected by numerous shared taxa. More taxa are shared at higher taxonomic levels including with humans, supporting their more ancestral common mammalian colonization followed by diversification. Phylogenies of selected oral bacterial lineages do not support stable human-dog microbial transfers but suggest diversification along mammalian lineages (apes and canids). Therefore, despite millennia of cohabitation and close interaction, the host and its native community controls and limits the assimilation of new microbes, even if closely related. Higher resolution metagenomic and microbial physiological studies, covering a larger mammalian diversity, should help understand how oral communities assemble, adapt, and interact with their hosts.

59 BASIC BIOLOGICAL SCIENCES↗

Clostridium autoethanogenum alters cofactor synthesis, redox metabolism, and lysine-acetylation in response to elevated H 2 :CO feedstock ratios for enhancing carbon capture efficiency

Clostridium autoethanogenum is an acetogenic bacterium that autotrophically converts carbon monoxide (CO) and carbon dioxide (CO 2 ) gases into bioproducts and fuels via the Wood–Ljungdahl pathway (WLP). To facilitate overall carbon capture efficiency, the reaction stoichiometry requires supplementation of hydrogen at an increased ratio of H 2 :CO to maximize CO 2 utilization; however, the molecular details and thus the ability to understand the mechanism of this supplementation are largely unknown. In order to elucidate the microbial physiology and fermentation where at least 75% of the carbon in ethanol comes from CO 2 , we established controlled chemostats that facilitated a novel and high (11:1) H 2 :CO uptake ratio. We compared and contrasted proteomic and metabolomics profiles to replicate continuous stirred tank reactors (CSTRs) at the same growth rate from a lower (5:1) H 2 :CO condition where ~ 50% of the carbon in ethanol is derived from CO 2 . Our hypothesis was that major changes would be observed in the hydrogenases and/or redox-related proteins and the WLP to compensate for the elevated hydrogen feed gas. Our analyses did reveal protein abundance differences between the two conditions largely related to reduction–oxidation (redox) pathways and cofactor biosynthesis, but the changes were more minor than we would have expected. While the Wood–Ljungdahl pathway proteins remained consistent across the conditions, other post-translational regulatory processes, such as lysine-acetylation, were observed and appeared to be more important for fine-tuning this carbon metabolism pathway. Metabolomic analyses showed that the increase in H 2 :CO ratio drives the organism to higher carbon dioxide utilization resulting in lower carbon storages and accumulated fatty acid metabolite levels. This research delves into the intricate dynamics of carbon fixation in C. autoethanogenum, examining the influence of highly elevated H 2 :CO ratios on metabolic processes and product outcomes. The study underscores the significance of optimizing gas feed composition for enhanced industrial efficiency, shedding light on potential mechanisms, such as post-translational modifications (PTMs), to fine-tune enzymatic activities and improve desired product yields.

09 BIOMASS FUELS↗

Whole genome resequencing data from a collection of Clostridium Thermocellum strains

Clostridium thermocellum is an anaerobic thermophilic bacterium that natively ferments cellulose to ethanol and organic acids. This data set is a collection of whole genome resequencing data for several hundred strains of Clostridium thermocellum. It includes strains that have been engineered to increase ethanol production, strains that have been engineered to understand microbial physiology, and strains that have been adapted for desired phenotypes including increased ethanol tolerance. Resequencing data consists of paired Illumina reads, 100-150 bp on each end, with a ~500 bp insert size. One data file containing raw Illumina data (interleaved) is available for each strain. We also provide data describing the mutations identified in each strain, and distinguish between inherited and newly observed mutations. In addition to resequencing data, we also provide metadata describing the lineage of each strain, and any targeted genetic modifications.

resequencing bio energy fermentation↗

FeGenie: a comprehensive tool for the identification of iron genes and iron gene neighborhoods in genomes and metagenome assemblies

Iron is a micronutrient for nearly all life on Earth. It can be used as an electron donor and electron acceptor by iron-oxidizing and iron-reducing microorganisms and is used in a variety of biological processes, including photosynthesis and respiration. While it is the fourth most abundant metal in the Earth’s crust, iron is often limiting for growth in oxic environments because it is readily oxidized and precipitated. Much of our understanding of how microorganisms compete for and utilize iron is based on laboratory experiments. However, the advent of next-generation sequencing and surge in publicly available sequence data has made it possible to probe the structure and function of microbial communities in the environment. To bridge the gap between our understanding of iron acquisition, iron redox cycling, iron storage, and magnetosome formation in model microorganisms and the plethora of sequence data available from environmental studies, we have created a comprehensive database of hidden Markov models (HMMs) based on genes related to iron acquisition, storage, and reduction/oxidation in Bacteria and Archaea. Along with this database, we present FeGenie, a bioinformatics tool that accepts genome and metagenome assemblies as input and uses our comprehensive HMM database to annotate provided datasets with respect to iron-related genes and gene neighborhood. An important contribution of this tool is the efficient identification of genes involved in iron oxidation and dissimilatory iron reduction, which have been largely overlooked by standard annotation pipelines. We validated FeGenie against a selected set of 28 isolate genomes and showcase its utility in exploring iron genes present in 27 metagenomes, 4 isolate genomes from human oral biofilms, and 17 genomes from candidate organisms, including members of the candidate phyla radiation. We show that FeGenie accurately identifies iron genes in isolates. Furthermore, analysis of metagenomes using FeGenie demonstrates that the iron gene repertoire and abundance of each environment is correlated with iron richness. While this tool will not replace the reliability of culture-dependent analyses of microbial physiology, it provides reliable predictions derived from the most up-to-date genetic markers. FeGenie’s database will be maintained and continually updated as new genes are discovered.

59 BASIC BIOLOGICAL SCIENCES↗

Culturing of “Unculturable” Subsurface Microbes: Natural Organic Carbon Source Fuels the Growth of Diverse and Distinct Bacteria From Groundwater

Recovery and cultivation of diverse environmentally-relevant microorganisms from the terrestrial subsurface remain a challenge despite recent advances in modern molecular technology. Here, we applied complex carbon (C) sources, i.e., sediment dissolved organic matter (DOM) and bacterial cell lysate, to enrich groundwater microbial communities for 30 days. As comparisons, we also included enrichments amended with simple C sources including glucose, acetate, benzoate, oleic acid, cellulose, and mixed vitamins. Our results demonstrate that complex C is far more effective in enriching diverse and distinct microorganisms from groundwater than simple C. Simple C enrichments yield significantly lower biodiversity, and are dominated by few phyla (e.g., Proteobacteria and Bacteroidetes ), while microcosms enriched with complex C demonstrate significantly higher biodiversity including phyla that are poorly represented in published culture collections (e.g., Verrucomicrobia , Planctomycetes , and Armatimonadetes ). Subsequent isolation from complex C enrichments yielded 228 bacterial isolates representing five phyla, 17 orders, and 56 distinct species, including candidate novel, rarely cultivated, and undescribed organisms. Results from this study will substantially advance cultivation and isolation strategies for recovering diverse and novel subsurface microorganisms. Obtaining axenic representatives of “once-unculturable” microorganisms will enhance our understanding of microbial physiology and function in different biogeochemical niches of terrestrial subsurface ecosystems.

59 BASIC BIOLOGICAL SCIENCES↗

Intensified Positive Arctic–Methane Feedback under IPCC Climate Scenarios in the 21st Century

The positive Arctic–methane (CH 4 ) feedback forms when more CH 4 is released from the Arctic tundra to warm the climate, further stimulating the Arctic to emit CH 4 . This study utilized the CLM-Microbe model to project CH 4 emissions across five distinct Arctic tundra ecosystems on the Alaska North Slope, considering three Shared Socioeconomic Pathway (SSP) scenarios using climate data from three climate models from 2016 to 2100. Employing a hyper-resolution of 5 m × 5 m within 40,000 m 2 domains accounted for the Arctic tundra’s high spatial heterogeneity; three sites were near $Utqia\dot{g}vik$ (US-Beo, US-Bes, and US-Brw), with one each in Atqasuk (US-Atq) and Ivotuk (US-Ivo). Simulated CH 4 emissions substantially increased by a factor of 5.3 to 7.5 under the SSP5–8.5 scenario compared to the SSP1–2.6 and SSP2–4.5 scenarios. The projected CH 4 emissions exhibited a stronger response to rising temperature under the SSP5–8.5 scenario than under the SSP1–2.6 and SSP2–4.5 scenarios, primarily due to strong temperature dependence and the enhanced precipitation-induced expansion of anoxic conditions that promoted methanogenesis. The CH 4 transport via ebullition and plant-mediated transport is projected to increase under all three SSP scenarios, and ebullition dominated CH 4 transport by 2100 across five sites. Projected CH 4 emissions varied in temperature sensitivity, with a Q 10 range of 2.7 to 60.9 under SSP1–2.6, 3.8 to 17.6 under SSP2–4.5, and 5.7 to 17.2 under SSP5–8.5. Compared with the other three sites, US-Atq and US-Ivo were estimated to have greater increases in CH 4 emissions due to warmer temperatures and higher precipitation. The fact that warmer sites and warmer climate scenarios had higher CH 4 emissions suggests an intensified positive Arctic–CH 4 feedback in the 21st century. Microbial physiology and substrate availability dominated the enhanced CH 4 production. The simulated intensified positive feedback underscores the urgent need for a more mechanistic understanding of CH 4 dynamics and the development of strategies to mitigate CH 4 across the Arctic.

54 ENVIRONMENTAL SCIENCES↗

Co-leveraging Scientific Advances in Space Biology and Astrobiology Towards Achieving NASA’s Life Science Objectives

Executive Summary: Distinct lines of scientific inquiry drives the separation of NASA’s fundamental life science research into Space Biology and Astrobiology. This division developed as a way to place life scientists alongside experts in the physical constraints that define the acclimation, adaptation and evolution of biology systems relevant to their respective subjects. For astrobiology, integration with disciplines such as geology, geochemistry, astronomy, planetary science, etc., enables a comprehensive assessment of the physical environment and its co-evolution with biological processes. Space Biology’s co-location with Physical Sciences places life science researchers adjacent to experts in the physical phenomena associated with microgravity and spaceflight, enabling an understanding of how the spaceflight environment affects biological systems. Despite this separation, aspects of both disciplines have converged on a similar, fundamental objective: to describe and understand the dynamics of complex living communities in the contexts of their physical environments. While the environmental systems and timescales are dramatically different, continuing to motivate the separation into distinct fields, similarities in the underlying objective present opportunities to find efficiencies, reduce overlap, and minimize duplication of effort. Space Biology and Astrobiology share a common need to understand microbial physiology in extreme environments – whether the ‘built’ spaceflight environment or the natural environments in which many astrobiology studies are conducted. In particular, open questions in each discipline require the development of quantitative frameworks, applicable at the ecosystem level, that support predictive capabilities for environments where observations are sparse. Additionally, both disciplines have a need to prepare, detect, and analyze the (potential) biological signal in complex samples-often in a completely autonomous fashion. The next decade will see NASA Space Biology moving to understand and describe the effects of the beyond low-earth orbit (BLEO) spaceflight environment on living systems. This new direction will dramatically reduce the opportunities for ground-based analysis of space-flown samples, driving space biology investigations towards fully autonomous experiments and missions. At the same time, astrobiology life detection missions aimed at detecting biosignatures on Mars and icy moons in the outer solar system could benefit from fully automated sample processing and analysis. There are opportunities to leverage instrument and method development between both disciplines within the context of these BLEO missions.

Astrobiology↗

On the use of antibiotics to reduce rhizoplane microbial populations in root physiology and ecology investigations

No straightforward method exists for separating the proportion of ion exchange and respiration due to rhizoplane microbial organisms from that of root ion exchange and respiration. We examined several antibiotics that might be used for the temporary elimination of rhizoplane bacteria from hydroponically grown wheat roots (Triticum aestivum cv. Veery 10). Each antibiotic was tested for herbicidal activity and plate counts were used to enumerate bacteria and evaluate antibiotic kinetics. Only lactam antibiotics (penicillins and cephalosporins) did not reduce wheat growth rates. Aminoglycosides, the pyrimidine trimethoprim, colistin and rifampicin reduced growth rates substantially. Antibiotics acted slowly, with maximum reductions in rhizoplane bacteria occurring after more than 48 h of exposure. Combinations of nonphytotoxic antibiotics reduced platable rhizoplane bacteria by as much as 98%; however, this was generally a reduction from about 10(9) to 10(6) colony forming units per gram of dry root mass, so that many viable bacteria remained on root surfaces. We present evidence which suggests that insufficient bacterial biomass exists on root surfaces of nonstressed plants grown under well-aerated conditions to quantitatively interfere with root nitrogen absorption measurements.

NASA Discipline Life Support Systems↗

Recovering new viruses from New Mexico soils

Here, we utilized metagenomic and size-filtered virome sequencing to recover 4,157 medium, high, or complete quality viral genomes from soils taken from three high elevation sites in New Mexico, USA. Among recovered viral genomes, 90% were from size-filtered samples, indicating the importance of this enrichment in assessments of complex viromes.

59 BASIC BIOLOGICAL SCIENCES↗

BONCAT-Live for isolation and cultivation of active environmental bacteria

In diverse environments, microbes drive a myriad of processes, from geochemical and nutrient cycling to interspecies interactions, including associations with plants and animals. Their physiological state is dynamic and impacted by abiotic and biotic conditions, responding to environmental fluctuations by changes in cellular metabolism, according to their genetic potential. Molecular, cellular, and genomic approaches can identify and measure microbial responses and adaptation to environmental changes in native communities. However, isolating individual microbial cells that respond to specific changes for cultivation has been difficult. To address this, we implemented a novel bacterial isolation approach (BONCAT-Live) by integrating bio-orthogonal non-canonical amino acid tagging (BONCAT) in diverse native communities, with isolation and cultivation of cells responding to specific stimuli, at different time scales. In frozen Arctic permafrost samples, we identified and isolated dormant bacteria that become active after thawing under native or nutrient-enriched conditions. From the Populus tree rhizosphere, we isolated strains that thrive under high concentrations of root exudates that act as defense compounds and nutrients. In the human microbiome, we identified and isolated bacteria that rapidly proliferated when exposed to metabolites provided by the host or other co-occurring microbes. Further characterization of isolated bacterial strains will provide opportunities for in-depth determination of how these microbes adapt to changes in their environments, individually and as part of model communities.

Analytical Methods↗

Harnessing evolution: leveraging bacterial isoprenoid pathway diversity toward improved bioengineering strategies

Isoprenoids play vital roles in all domains of life, from beta-carotene in bacteria to heme in humans. Two distinct metabolic pathways have evolved to synthesize the critical precursor of all mature isoprenoids: the mevalonate (MEV) and the methylerythritol phosphate (MEP) pathways. Here, we quantify the extensive inter- and intra-genus heterogeneity in the usage of these two pathways with particular emphasis on rare bacteria that encode both, or neither, pathways. Furthermore, MEP intermediates themselves have non-isoprenogenic roles that may underlie evolutionary pressures driving pathway diversification. Understanding isoprenoid biosynthesis in bacteria offers new avenues toward more sustainable engineering of economically relevant molecules in microbes.

Biotechnology and Synthetic Biology↗

Composition and physiological profiling of sprout-associated microbial communities

The native microfloras of various types of sprouts (alfalfa, clover, sunflower, mung bean, and broccoli sprouts) were examined to assess the relative effects of sprout type and inoculum factors (i.e., sprout-growing facility, seed lot, and inoculation with sprout-derived inocula) on the microbial community structure of sprouts. Sprouts were sonicated for 7 min or hand shaken with glass beads for 2 min to recover native microfloras from the surface, and the resulting suspensions were diluted and plated. The culturable fraction was characterized by the density (log CFU/g), richness (e.g., number of types of bacteria), and diversity (e.g., microbial richness and evenness) of colonies on tryptic soy agar plates incubated for 48 h at 30 degrees C. The relative similarity between sprout-associated microbial communities was assessed with the use of community-level physiological profiles (CLPPs) based on patterns of utilization of 95 separate carbon sources. Aerobic plate counts of 7.96 +/- 0.91 log CFU/g of sprout tissue (fresh weight) were observed, with no statistically significant differences in microbial cell density, richness, or diversity due to sprout type, sprout-growing facility, or seed lot. CLPP analyses revealed that the microbial communities associated with alfalfa and clover sprouts are more similar than those associated with the other sprout types tested. Variability among sprout types was more extensive than any differences between microbial communities associated with alfalfa and clover sprouts from different sprout-growing facilities and seed lots. These results indicate that the subsequent testing of biocontrol agents should focus on similar organisms for alfalfa and clover, but alternative types may be most suitable for the other sprout types tested. The inoculation of alfalfa sprouts with communities derived from various sprout types had a significant, source-independent effect on microbial community structure, indicating that the process of inoculation alters the dynamics of community development regardless of the types of organisms involved.

NASA Center KSC↗

Nitrogen fixation and resorption efficiency differences among twelve upland and lowland switchgrass cultivars

In nitrogen (N)-limited terrestrial ecosystems, plants employ various strategies to acquire and conserve N, including translocation of N in perennial tissues and stimulation of N fixation in roots and soils. Switchgrass (Panicum virgatum L.) is a genotypically and phenotypically diverse perennial grass with two distinct ecotypes (lowland and upland) and numerous genotypes. It grows well in low-N soils, likely because of its ability to translocate N and to associate with N-fixing microbes, but little is known about variation in these traits among cultivars or even ecotypes. We measured N translocation, N fixation potential in roots and soils, soil net N mineralization, soil net nitrification, and biomass yields in 12 switchgrass cultivars grown in a replicated block experiment in southwestern Michigan, USA. Lowland cultivars had higher yields, rates of N translocation, soil net N mineralization, and N fixation potentials on washed, non-sterile roots, while upland cultivars exhibited higher N fixation potentials in root-free soil. N resorption efficiencies averaged 53±5 [SE] % for lowland versus 29±3% for upland cultivars. Additionally, there were significant among-cultivar differences for all response variables except mineralization and nitrification, with differences likely explained by cultivar-specific physiologies and microbial communities. Here, the ideal cultivar for biofuels is one that can maintain high yields with minimal fertilizer addition and there appear to be several cultivars that meet these criteria. In addition, results suggest substantial N cycle differences among cultivars that might be exploited by breeders to create new or improved high-yielding, N-conserving switchgrass lines.

59 BASIC BIOLOGICAL SCIENCES↗

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↗

Transcriptomic data sets examining several stress responses in Zymomonas mobilis strains

We used RNA-seq to compare gene expression from Zymomonas mobilis ZM4 grown under various conditions: aerobic ± paraquat, anaerobic ± hydrogen peroxide, or an iron chelator. We analyzed two mutant strains lacking predicted transcription factors (ZMO_0442 and ZMO_1411) grown under aerobic or anaerobic conditions. We report the RNA-seq data from these experiments.

Bacterial Stress Response↗