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At least 271 records · Page 15

Recruiting Human Microbiome Shotgun Data to Site-Specific Reference Genomes

The human body consists of innumerable multifaceted environments that predispose colonization by a number of distinct microbial communities, which play fundamental roles in human health and disease. In addition to community surveys and shotgun metagenomes that seek to explore the composition and diversity of these microbiomes, there are significant efforts to sequence reference microbial genomes from many body sites of healthy adults. To illustrate the utility of reference genomes when studying more complex metagenomes, we present a reference-based analysis of sequence reads generated from 55 shotgun metagenomes, selected from 5 major body sites, including 16 sub-sites. Interestingly, between 13% and 92% (62.3% average) of these shotgun reads were aligned to a then-complete list of 2780 reference genomes, including 1583 references for the human microbiome. However, no reference genome was universally found in all body sites. For any given metagenome, the body site-specific reference genomes, derived from the same body site as the sample, accounted for an average of 58.8% of the mapped reads. While different body sites did differ in abundant genera, proximal or symmetrical body sites were found to be most similar to one another. The extent of variation observed, both between individuals sampled within the same microenvironment, or at the same site within the same individual over time, calls into question comparative studies across individuals even if sampled at the same body site. This study illustrates the high utility of reference genomes and the need for further site-specific reference microbial genome sequencing, even within the already well-sampled human microbiome.

59 BASIC BIOLOGICAL SCIENCES↗

Coupling Genetic and Chemical Microbiome Profiling Reveals Heterogeneity of Archaeome and Bacteriome in Subsurface Biofilms That Are Dominated by the Same Archaeal Species

Earth harbors an enormous portion of subsurface microbial life, whose microbiome flux across geographical locations remains mainly unexplored due to difficult access to samples. Here, we investigated the microbiome relatedness of subsurface biofilms of two sulfidic springs in southeast Germany that have similar physical and chemical parameters and are fed by one deep groundwater current. Due to their unique hydrogeological setting these springs provide accessible windows to subsurface biofilms dominated by the same uncultivated archaeal species, called SM1 Euryarchaeon. Comparative analysis of infrared imaging spectra demonstrated great variations in archaeal membrane composition between biofilms of the two springs, suggesting different SM1 euryarchaeal strains of the same species at both aquifer outlets. This strain variation was supported by ultrastructural and metagenomic analyses of the archaeal biofilms, which included intergenic spacer region sequencing of the rRNA gene operon. At 16S rRNA gene level, PhyloChip G3 DNA microarray detected similar biofilm communities for archaea, but site-specific communities for bacteria. Both biofilms showed an enrichment of different deltaproteobacterial operational taxonomic units, whose families were, however, congruent as were their lipid spectra. Consequently, the function of the major proportion of the bacteriome appeared to be conserved across the geographic locations studied, which was confirmed by dsrB-directed quantitative PCR. Consequently, microbiome differences of these subsurface biofilms exist at subtle nuances for archaea (strain level variation) and at higher taxonomic levels for predominant bacteria without a substantial perturbation in bacteriome function. The results of this communication provide deep insight into the dynamics of subsurface microbial life and warrant its future investigation with regard to metabolic and genomic analyses.

54 ENVIRONMENTAL SCIENCES↗

Lost and Found: Rediscovering Microbiome-Associated Phenotypes that Reshape Agricultural Sustainability

Modern agriculture faces an urgent need to improve nutrient use efficiency while reducing environmental impacts. Here, we show that ancestral traits controlling rhizosphere microbiome functions can be reintroduced into elite maize through targeted teosinte introgressions. Using near-isogenic lines, we mapped microbiome-associated phenotypes (MAPs) derived from teosinte that suppress nitrification and denitrification—key microbial processes contributing to nitrogen loss. These introgressions altered root exudate chemistry, resulting in distinct microbial assemblies and enhanced nitrogen retention. We identified candidate loci and metabolites responsible for suppressive activity and demonstrated their functional effects in vitro. Our findings reveal a genetic and biochemical basis for rewilding microbiome-mediated ecosystem services in crops, offering a scalable path toward sustainable nutrient management in global agriculture. ---- These maize root exduate metabolomics data are a subset of this larger project and make up a phenotyping for candidate lines.

Favela, Alonso [School of Plant Sciences, Universi↗

Nitrogen fertilizer affects the Populus microbiome

Plants are colonized by numerous microorganisms serving important symbiotic functions that aid in plant growth and success. Populus deltoides (Eastern Cottonwood) and Populus trichocarpa (Black Cottonwood) are potential biofuel crops that rely on microbial symbionts to complete essential tasks, such as nitrogen fixation. Often, in agricultural and biofuel production, exogenous nitrogen is added to soils, since nitrogen availability is the limiting factor for plant growth. However, these additions may alter the soil microbial communities and result in unintentional consequences if excess nitrogen escapes the setting. We sought to determine how nitrogen fertilization alters the microbial communities within the soil, roots, and leaves of two cottonwoods. We grew P. deltoides and P. trichocarpa in a greenhouse with and without added nitrogen fertilizer, in two soil types. The soils were from Oregon and West Virginia and had distinctive chemical properties and different initial soil microbial communities. We hypothesized that the abundance of diazotrophs within the soils would increase as nitrogen became limiting. Over the 2-month study, we assessed plant growth characteristics for over 160 Populus plants. We investigated the root, leaf, and soil microbiomes of selected plants using 16S rRNA gene and ITS2 PCR amplicon DNA sequencing. We then used bioinformatics tools to evaluate the composition and diversity of these microbiomes. The microbiomes differed significantly across all three experimental factors - nitrogen addition, Populus species, and soil type.

09 BIOMASS FUELS↗

BSSD Performance Metric report: LLNL Soil Microbiome SFA (Q1 2021)

The LLNL “Microbes Persist” Soil Microbiome Scientific Focus Area (SFA) seeks to determine how microbial soil ecophysiology, population dynamics, and microbe-mineral-organic matter interactions regulate the persistence of microbial residues and the formation of soil carbon. Our SFA research program is now four years old; it evolved and benefited from previously-funded BSSD projects in the Firestone (UCB), Banfield (UCB), Sullivan (OSU) and Hungate (NAU) labs. We use stable isotope probing in combination with ‘omics to measure how changing water regimes shape activity of individual microbial populations and ecophysiological traits that affect the fate of microbial and plant C. Using measures of population dynamics and microbiome-mineral interactions, we are working to synthesize both genomescale and ecosystem-scale models of soil organic matter (SOM) turnover, to predict the long-aspired connection between soil microbiomes and fate of soil C.

54 ENVIRONMENTAL SCIENCES↗

Expanding the Scope of Genomic Security: Targeted Genome Editing within Microbiomes through Designer Bacteriophage Vectors

The ability to engineer the genome of a bacterial strain, not as an isolate, but while present among other microbes in a microbiome, would open new technological possibilities in the areas of medicine, energy and biomanufacturing. Our approach is to develop sets of phages (bacterial viruses) active on the target strain and themselves engineered to act not as killers but as vectors for gene delivery. This approach is rooted in our bioinformatic tools that map prophages accurately within bacterial genomes. We present new bioinformatic results in cross-contig search, design of phage genome assemblies, satellites that embed within prophages, alignment of large numbers of biological sequences, and improvement of reference databases for prophage discovery. We targeted a Pseudomonas putida strain within a lignin-degrading microbiome, but were unable to obtain active phages, and turned toward a defined microbiome of the mouse gut.

59 BASIC BIOLOGICAL SCIENCES↗

PNNL Soil Microbiome SFA

PNNL's Soil Microbiome Science Focus Area (SFA) is focused on understanding the basic biology underpinning how interactions among various soil microbial community members, across trophic levels, lead to the emergence of community functions. Moisture, in particular, drives microbial interactions and influences everything from cell function to substrate fate within soils. The group predicts this results in repeatable, predictable phenotypes. The sum of these phenotypes comprises the “soil metaphenome”. Understanding how the soil metaphenome shifts in response to moisture will provide a basis for modeling and predicting these shifts in reaction network responses. PNNL’s Soil Microbiome SFA project repository on DataHub allows for exploring and downloading integrated experimental omics data, experimental metadata, pre- and post-processed data files, and other associated materials directly related to experimental project publication data. Visit the PNNL Soil Microbiome Science Focus Area Program homepage for more information.

54 ENVIRONMENTAL SCIENCES↗

Landscape Topography and Regional Drought Alters Dust Microbiomes in the Sierra Nevada of California

Dust provides an ecologically significant input of nutrients, especially in slowly eroding ecosystems where chemical weathering intensity limits nutrient inputs from underlying bedrock. In addition to nutrient inputs, incoming dust is a vector for dispersing dust-associated microorganisms. While little is known about dust-microbial dispersal, dust deposits may have transformative effects on ecosystems far from where the dust was emitted. Using molecular analyses, we examined spatiotemporal variation in incoming dust microbiomes along an elevational gradient within the Sierra Nevada of California. We sampled throughout two dry seasons and found that dust microbiomes differed by elevation across two summer dry seasons (2014 and 2015), which corresponded to competing droughts in dust source areas. Dust microbial taxa richness decreased with elevation and was inversely proportional to dust heterogeneity. Likewise, dust phosphorus content increased with elevation. At lower elevations, early season dust microbiomes were more diverse than those found later in the year. The relative abundances of microbial groups shifted during the summer dry season. Furthermore, mutualistic fungal diversity increased with elevation, which may have corresponded with the biogeography of their plant hosts. Although dust fungal pathogen diversity was equivalent across elevations, elevation and sampling month interactions for the relative abundance, diversity, and richness of fungal pathogens suggest that these pathogens differed temporally across elevations, with potential implications for humans and wildlife. This study shows that landscape topography and droughts in source locations may alter the composition and diversity of ecologically relevant dust-associated microorganisms.

59 BASIC BIOLOGICAL SCIENCES↗

Interrogating the role of the milk microbiome in mastitis in the multi-omics era

There is growing interest in a functional understanding of milk-associated microbiota as there is ample evidence that host-associated microbial communities play an active role in host health and phenotype. Mastitis, characterized by painful inflammation of the mammary gland, is prevalent among lactating humans and agricultural animals and is associated with significant clinical and economic consequences. The etiology of mastitis is complex and polymicrobial and correlative studies have indicated alterations in milk microbial community composition. Recent evidence is beginning to suggest that a causal relationship may exist between the milk microbiota and host phenotype in mastitis. Multi-omic approaches can be leveraged to gain a mechanistic, molecular level understanding of how the milk microbiome might modulate host physiology, thereby informing strategies to prevent and ameliorate mastitis. In this paper, we review existing studies that have utilized omics approaches to investigate the role of the milk microbiome in mastitis. We also summarize the strengths and challenges associated with the different omics techniques including metagenomics, metatranscriptomics, metaproteomics, metabolomics and lipidomics and provide perspective on the integration of multiple omics technologies for a better functional understanding of the milk microbiome.

metabolomics↗

Editorial for Special Issue “Advances in Soil Microbiome”

The soil microbiome (the community of all soil microorganisms and their surrounding environment) is a critical part of our ecological network. These microbial communities carry out several important functions, including the promotion of plant growth and the cycling of carbon and nitrogen sources. Despite the importance of this community, and the recent advances in multi-omic analysis and other methods designed to query complex systems, there is a still a large knowledge gap regarding the soil microbiome, its response to changing climates and moisture levels, the specific molecular mechanisms that drive its role in the environment, and how these questions can be answered through the use of model systems. Here, we present eleven new papers that seek to answer these questions and expand our knowledge of the soil microbiome.

59 BASIC BIOLOGICAL SCIENCES↗

Method Development for In Situ Microbiome Profiling of the Water Recovery System’s Wastewater Tank Onboard the International Space Station

A distinctive microbial community has inhabited the wastewater tank within the International Space Station Water Recovery System (WRS) for over 14 years and experienced the stressors associated with the microgravity environment. The WRS generates potable water for the crew from urine distillate, humidity condensate, Sabatier product water, and the occasional off-loading of ground-supplied water. The reservoir for these products, the wastewater tank, does not have a means of microbial control. While samples are occasionally collected for analysis, the time between sample collection and the return to Earth, as well as the lack of preservation, results in a skewed depiction of the microbiome. Routinely observed from these returned samples are high counts (105 – 106 colony forming units per mL) and two prevailing genera, Burkholderia and Ralstonia. The wastewater tank likely contains a more diverse microbiome, as a higher diversity of bacteria and fungus has been noted upstream and downstream of the tank. To characterize the microbial profile of the tank, analysis needs to occur at the time of sample collection. Toward this goal, a method for in situ analysis based on nanopore sequencing was developed. The filter-to-sequencer method evolved from previous work that has been validated onboard the ISS (BEST payload and the BioMole Crew Health Care Systems Facility). The method, including filtration, DNA extraction, purification, amplification, library preparation, and nanopore sequencing will be described. Additionally, data collected with this method from both ISS and terrestrial samples will be detailed. The consumables needed to support in situ analysis of the tank are set to the launch to the ISS in the spring of 2023. This investigation will allow for the first accurate characterization of the microbiome of the tank providing insight for crew health, planetary protection, and has the potential to enable engineering controls for future space station water systems.

Sarah Stahl-Rommel↗

Impacts of Atmospheric Microbiomes Field Campaign Report

Aerosols of microbial origin (e.g., bacteria, fungi, etc.; also known as “atmospheric microbiome”) are ubiquitous in the atmosphere where they can remain suspended for several days to weeks and be transported over long ranges across continents. There is growing recognition that a significant fraction of total particulates suspended in the atmosphere may host microorganisms. Studies indicate that atmospheric microbial transport can play a significant role in processes associated with climate change and atmospheric chemistry. Furthermore, microbe-bearing particles are continually exchanged between ecosystems through global atmospheric transport pathways with the potential to influence the dispersion, composition, and dynamics of microbial communities across diverse ecosystems, including land managed, modified, and used by humans. The impacts of these atmospheric microbiomes on climate are poorly understood. The goal of this work is to establish fundamental new capabilities that enable prediction of atmospheric microbiome dynamics and interactions from source to final deposition. To achieve these objectives, surface soils (0-5 cm depth) were taken from within a 2.5-m radial area near the likely origin of the airborne particulates. In addition, we collected bulk and size-resolved airborne samples from a sampling line located ~4 m above the central U.S. Department of Energy (DOE) Atmospheric Radiation Measurement (ARM) Southern Great Plains (SGP) site from 25/10 to 31/10 in 2016. The SGP site is located in north-central Oklahoma and experiences a broad range of air mass, cloud, and atmospheric state disturbances. The site is surrounded by various agricultural crops and pasture fields. In total, 20 samples were collected over one week in October 2016, and each sample collection period was again approximately 12 hours. At both locations, the samples were collected on PTFE laminated membrane 0.45-μm filters (Sterlitech Corp.) mounted inside a 90-mm single-stage filter assembly (Savillex Inc.). The air was sampled from the sampling inlet located ~4 m height from the ground at total ~0.23 m3/min, which allowed us to sample total ~166 m3 of air per filter. The filter holder assembly and membrane filter holder tips were cleaned with isopropyl alcohol before a new filter was loaded. Blank filters were also collected to understand the background bacterial loading and composition but without operating the filter assembly. Both ambient air collected and blank filters were stored and shipped frozen at ~-20 °C to avoid the contamination and growth of unwanted bacterial communities. In parallel, we also collected the samples using a Micro-Orifice Uniform Deposit Impactor (MOUDI) for microscopy analysis.

54 ENVIRONMENTAL SCIENCES↗

The Oral and Skin Microbiomes of Captive Komodo Dragons Are Significantly Shared with Their Habitat

Animals, including humans, have evolved in the context of exposure to a variety of microbial organisms present in the environment. Only recently have humans, and some animals, begun to spend a significant amount of time in enclosed artificial environments, rather than in the more natural spaces in which most of evolution took place. The consequences of this radical change in lifestyle likely extend to the microbes residing in and on our bodies and may have important implications for health and disease. A full characterization of host-microbe sharing in both closed and open environments will provide crucial information that may enable the improvement of health in humans and in captive animals, both of which experience a greater incidence of disease (including chronic illness) than counterparts living under more ecologically natural conditions. Examining the way in which animals, including those in captivity, interact with their environment is extremely important for studying ecological processes and developing sophisticated animal husbandry. Here we use the Komodo dragon (Varanus komodoensis) to quantify the degree of sharing of salivary, skin, and fecal microbiota with their environment in captivity. Both species richness and microbial community composition of most surfaces in the Komodo dragon’s environment are similar to the Komodo dragon’s salivary and skin microbiota but less similar to the stool-associated microbiota. We additionally compared host-environment microbiome sharing between captive Komodo dragons and their enclosures, humans and pets and their homes, and wild amphibians and their environments. We observed similar host-environment microbiome sharing patterns among humans and their pets and Komodo dragons, with high levels of human/pet- and Komodo dragon-associated microbes on home and enclosure surfaces. In contrast, only small amounts of amphibian-associated microbes were detected in the animals’ environments. We suggest that the degree of sharing between the Komodo dragon microbiota and its enclosure surfaces has important implications for animal health. These animals evolved in the context of constant exposure to a complex environmental microbiota, which likely shaped their physiological development; in captivity, these animals will not receive significant exposure to microbes not already in their enclosure, with unknown consequences for their health. Animals, including humans, have evolved in the context of exposure to a variety of microbial organisms present in the environment. Only recently have humans, and some animals, begun to spend a significant amount of time in enclosed artificial environments, rather than in the more natural spaces in which most of evolution took place. The consequences of this radical change in lifestyle likely extend to the microbes residing in and on our bodies and may have important implications for health and disease. A full characterization of host-microbe sharing in both closed and open environments will provide crucial information that may enable the improvement of health in humans and in captive animals, both of which experience a greater incidence of disease (including chronic illness) than counterparts living under more ecologically natural conditions.

59 BASIC BIOLOGICAL SCIENCES↗

The Earth Microbiome project: successes and aspirations

Here, the Earth Microbiome Project (EMP) was launched in August 2010, with the ambitious aim of constructing a global catalogue of the uncultured microbial diversity of this planet. The primary vision of the Earth Microbiome Project, to process the microbial diversity and functional potential from approximately 200,000 environmental samples, marks it as an undertaking so massive that it was at first considered to be pure folly (as late as 2012, Jonathan Eisen was quoted in Nature as saying ‘Knight and Gilbert literally talk about sampling the entire planet. It is ludicrous and not feasible - yet they are doing it’ [1]).

59 BASIC BIOLOGICAL SCIENCES↗

Understanding Cultivar-Specificity and Soil Determinants of the Cannabis Microbiome

Understanding microbial partnerships with the medicinally and economically important crop Cannabis has the potential to affect agricultural practice by improving plant fitness and production yield. Furthermore, Cannabis presents an interesting model to explore plant-microbiome interactions as it produces numerous secondary metabolic compounds. Here we present the first description of the endorhiza-, rhizosphere-, and bulk soil-associated microbiome of five distinct Cannabis cultivars. Bacterial communities of the endorhiza showed significant cultivar-specificity. When controlling cultivar and soil type the microbial community structure was significantly different between plant cultivars, soil types, and between the endorhiza, rhizosphere and soil. In conclusion, the influence of soil type, plant cultivar and sample type differentiation on the microbial community structure provides support for a previously published two-tier selection model, whereby community composition across sample types is determined mainly by soil type, while community structure within endorhiza samples is determined mainly by host cultivar.

54 ENVIRONMENTAL SCIENCES↗

Simple Analysis of Primary and Secondary Bile Salt Hydrolysis in Mouse and Human Gut Microbiome Samples by Using Fluorogenic Substrates

Animals produce bile to maximize absorption of lipophilic nutrients in the gut. The physical properties of bile are largely dictated by amphipathic bile salt molecules, which also act as signaling molecules that bind host receptors and modulate physiological processes. Upon excretion of bile salts into the gut, the gut microbiome can create metabolites that have modified signaling capabilities. The category and magnitude of bile salt metabolism can affect the host either positively or negatively. A key modification is bile salt hydrolysis, which is a prerequisite step for all additional microbial modification. Here, we have synthesized five different fluorogenic bile salts that report on hydrolysis activity using a simple, continuous assay. We profile activity in both murine and human fecal samples. Our data demonstrate that most gut microbiomes have the highest capacity for hydrolysis of host-produced bile salts, but some microbially modified bile salts also display significant turnover.

59 BASIC BIOLOGICAL SCIENCES↗

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↗

Not a “they” but a “we”: The microbiome helps promote our well-being

Anaerobic microbes in the human gut produce beneficial and harmful compounds, as well as neutral compounds like trimethylamine, which undergoes microbial metabolism or host-catalyzed transformation into proatherogenic trimethylamine-N-oxide. Ellenbogen et al. identified a microbiome-associated demethylase that short-circuits the production of trimethylamine-N-oxide from the metabolite γ-butyrobetaine and instead produces methyltetrahydrofolate, a key intermediate in the microbial production of beneficial small-chain fatty acids. This article highlights an example of how the microbiome is integrally involved in producing metabolites that support our health and in preventing the formation of compounds that promote disease.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗