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At least 55 records · Page 3

Metagenomics Reveals Pervasive Bacterial Populations and Reduced Community Diversity across the Alaska Tundra Ecosystem

How soil microbial communities contrast with respect to taxonomic and functional composition within and between ecosystems remains an unresolved question that is central to predicting how global anthropogenic change will affect soil functioning and services. In particular, it remains unclear how small-scale observations of soil communities based on the typical volume sampled (1-2 g) are generalizable to ecosystem-scale responses and processes. This is especially relevant for remote, northern latitude soils, which are challenging to sample and are also thought to be more vulnerable to climate change compared to temperate soils. Here, we employed well-replicated shotgun metagenome and 16S rRNA gene amplicon sequencing to characterize community composition and metabolic potential in Alaskan tundra soils, combining our own datasets with those publically available from distant tundra and temperate grassland and agriculture habitats. We found that the abundance of many taxa and metabolic functions differed substantially between tundra soil metagenomes relative to those from temperate soils, and that a high degree of OTU-sharing exists between tundra locations. Tundra soils were an order of magnitude less complex than their temperate counterparts, allowing for near-complete coverage of microbial community richness (~92% breadth) by sequencing, and the recovery of 27 high-quality, almost complete ( > 80% completeness) population bins. These population bins, collectively, made up to ~10% of the metagenomic datasets, and represented diverse taxonomic groups and metabolic lifestyles tuned toward sulfur cycling, hydrogen metabolism, methanotrophy, and organic matter oxidation. Several population bins, including members of Acidobacteria, Actinobacteria, and Proteobacteria, were also present in geographically distant (~100-530 km apart) tundra habitats (full genome representation and up to 99.6% genome-derived average nucleotide identity). Collectively, our results revealed that Alaska tundra microbial communities are less diverse and more homogenous across spatial scales than previously anticipated, and provided DNA sequences of abundant populations and genes that would be relevant for future studies of the effects of environmental change on tundra ecosystems.

climate change↗

Wave energy in season: a comparative approach to feasibility of seasonal deployments for remote coastal communities

Remote coastal communities, which could be early adopters of wave energy projects, have concerns over costs, conflicts, and potential risks of development. Designers and developers are challenged to address these community concerns as they continue to develop wave energy technologies. One potential means of reducing costs, conflicts, and risks, especially for demonstration and pilot-scale projects, could be planning a deployment that operates for only a portion of the year—a seasonal deployment. Here, in this paper, we examine the impacts of a seasonal deployment in terms of cost, electricity production, operations and maintenance, environmental impacts, and community benefits. We take a holistic, comparative approach to feasibility that can be replicated for other comparative studies. We estimate electricity production using a point absorber WEC modeled near Sitka, AK, USA and optimized for the given sea conditions. We determine that, for remote community sized projects, seasonal deployments could result in small cost savings (less than 10 %), but larger decreases in annual energy production (around 30 % for our case study area). Seasonal deployments could be preferable in places with seasonal energy needs, if failures and device access become a major hindrance to wave energy technology development, or as a cautionary approach to introducing new technology to the oceans. We also determine that a highly seasonal wave resource is not necessarily a requirement for seasonal deployments to be considered. Seasonal deployments are an alternative to year-round deployments that can be considered in places where marine spatial conflict is a seasonal concern.

WEC optimizations↗

Context dependent fungal and bacterial soil community shifts in response to recent wildfires in the Southern Appalachian Mountains

Decades of fire suppression coupled with changing climatic conditions have increased the frequency and intensity of wildfires. Here, the Southern Appalachia region of the United States is predicted to be particularly susceptible to climatic changes, with predicted increases in fire severity and occurrence. Following the record breaking fire season in 2016 in Southern Appalachia, we examined wildfire impacts on soil chemistry and below ground communities (fungi and bacteria – Illumina MiSeq) within two substrates (duff and soil) at two adjacent locations with similar plant communities (Great Smoky Mountains National Park – ‘Chimney Top 2’ Fire (GRSM) and Nantahala National Forest – ‘Cliffside’ Fire (NNF)) from replicate plots representing a range of fire severities (unburned, low severity, moderate severity, severe). Differing fire severities changed community composition, and fire severity played a stronger role in structuring bacterial communities than in structuring fungal communities. Further, fire impacts on soil communities and functional guilds responses were location- and substrate-specific with NNF responding more strongly to fire than GRSM. Additionally, using a novel analysis tool (Axis Weighted Ordination Distance – AWOrD), domain and location specific responses to wildfire severity are demonstrated. Taken together, our results suggest context-dependency in microbial responses to fire that must be accounted for to generate ecosystem-wide recovery predictions.

54 ENVIRONMENTAL SCIENCES↗

Tundra Soil Viruses Mediate Responses of Microbial Communities to Climate Warming

The rise of global temperature causes the degradation of the substantial reserves of carbon (C) stored in tundra soils, in which microbial processes play critical roles. Viruses are known to influence the soil C cycle by encoding auxiliary metabolic genes and infecting key microorganisms, but their regulation of microbial communities under climate warming remains unexplored. In this study, we evaluated the responses of viral communities for about 5 years of experimental warming at two depths (15 to 25 cm and 45 to 55 cm) in the Alaskan permafrost region. Our results showed that the viral community and functional gene composition and abundances (including viral functional genes related to replication, structure, infection, and lysis) were significantly influenced by environmental conditions such as total nitrogen (N), total C, and soil thawing duration. Although long-term warming did not impact the viral community composition at the two depths, some glycoside hydrolases encoded by viruses were more abundant at both depths of the warmed plots. With the continuous reduction of total C, viruses may alleviate methane release by altering infection strategies on methanogens. Importantly, viruses can adopt lysogenic and lytic lifestyles to manipulate microbial communities at different soil depths, respectively, which could be one of the major factors causing the differences in microbial responses to warming. This study provides a new ecological perspective on how viruses regulate the responses of microbes to warming at community and functional scales.

54 ENVIRONMENTAL SCIENCES↗

A novel analysis method for paired-sample microbial ecology experiments

Many microbial ecology experiments use sequencing data to measure a community s response to an experimental treatment. In a common experimental design, two units, one control and one experimental, are sampled before and after the treatment is applied to the experimental unit. The four resulting samples contain information about the dynamics of organisms that respond to the treatment, but there are no analytical methods designed to extract exactly this type of information from this configuration of samples. Here we present an analytical method specifically designed to visualize and generate hypotheses about microbial community dynamics in experiments that have paired samples and few or no replicates. The method is based on the Poisson lognormal distribution, long studied in macroecology, which we found accurately models the abundance distribution of taxa counts from 16S rRNA surveys. To demonstrate the method s validity and potential, we analyzed an experiment that measured the effect of crude oil on ocean microbial communities in microcosm. Our method identified known oil degraders as well as two clades, Maricurvus and Rhodobacteraceae, that responded to amendment with oil but do not include known oil degraders. Furthermore, our approach is sensitive to organisms that increased in abundance only in the experimental unit but less sensitive to organisms that increased in both control and experimental units, thus mitigating the role of bottle effects .

59 BASIC BIOLOGICAL SCIENCES↗

Form Follows Function (F3): A framework for community-based energy resilience planning in the Midwest

This is the final technical report for the Form Follows Function (F3) grant project led by the City of Duluth, MN. F3 was an innovative two-year initiative that created a replicable, human-centered energy resilience planning process specifically designed for smaller, cold-climate communities in the Midwest. F3 demonstrated both systems-level government planning and community-based research techniques to inform renewable investments and community preparedness strategies.

14 SOLAR ENERGY↗

Phosphate amendment drives bloom of RNA viruses after soil wet-up

Soil rewetting after a dry period results in a surge of activity and succession in both microbial and DNA virus communities. Less is known about the response of RNA viruses to soil rewetting—while they are highly diverse and widely distributed in soil, they remain understudied. We hypothesized that RNA viruses would show temporal succession following rewetting and that phosphate amendment would influence their trajectory, as viral proliferation may cause phosphorus limitation. Using 39 time-resolved metatranscriptomes and amplicon data, 2190 RNA viral populations were identified across five phyla, with 26 % of these predicted to infect bacteria, and 11 % fungi. Only 1.2 % of viral populations had annotated capsid genes, suggesting most persist via intracellular replication without a free virion phase. Phosphate amendment altered RNA viral community composition within the first week and amended vs. unamended communities remained distinguishable for up to three weeks. While the overall host community remained stable, certain bacterial populations showed reduced abundance in phosphate-amended soils, likely due to increased viral lysis, as RNA bacteriophages proliferated significantly. Notably, 60 % of the viruses with increased abundance under phosphate amendment belonged to basal Lenarviricota clades rather than well-known groups like Leviviricetes. We estimate RNA bacteriophage infections may affect 10 7 –10 9 bacteria per gram of soil, aligning with the total bacterial population (10 7 –10 10 g -1 soil), suggesting that RNA phages significantly influence bacterial communities post-wet-up, with phosphorus availability modulating this effect.

59 BASIC BIOLOGICAL SCIENCES↗

Putative Mixotrophic Nitrifying-Denitrifying Gammaproteobacteria Implicated in Nitrogen Cycling Within the Ammonia/Oxygen Transition Zone of an Oil Sands Pit Lake

Anthropogenically-impacted environments offer the opportunity to discover novel microbial species and metabolisms, which may be undetectable in natural systems. Here, a combined metagenomic and geochemical study in Base Mine Lake, Alberta, Canada, which is the only oil sands end pit lake to date, revealed that nitrification was performed by members from Nitrosomonadaceae, Chloroflexi and unclassified Gammaproteobacteria "MBAE14." While Nitrosomonadaceae and Chloroflexi groups were relatively abundant in the upper oxygenated zones, MBAE14 dominated the hypoxic hypolimnetic zones (approximately 30% of total microbial communities); MBAE14 was not detected in the underlying anoxic tailings. Replication rate analyses indicate that MBAE14 grew in metalimnetic and hypolimnetic water cap regions, most actively at the metalimnetic, ammonia/oxygen transition zone consistent with it putatively conducting nitrification. Detailed genomic analyses of MBAE14 evidenced both ammonia oxidation and denitrification into dinitrogen capabilities. However, the absence of known CO 2 -fixation genes suggests a heterotrophic denitrifying metabolism. Functional marker genes of ammonia oxidation ( amo and hao ) in the MBAE14 genome are homologous with those conserved in autotrophic nitrifiers, but not with those of known heterotrophic nitrifiers. We propose that this novel MBAE14 inhabits the specific ammonia-rich, oxygen and labile organic matter-limited conditions occurring in Base Mine Lake which selectively favors mixotrophic coupled nitrifier denitrification metabolism. Our results highlight the opportunities to better constrain biogeochemical cycles from the application of metagenomics to engineered systems associated with extractive resource sectors.

59 BASIC BIOLOGICAL SCIENCES↗

Selection, Succession, and Stabilization of Soil Microbial Consortia

Soil microorganisms play fundamental roles in cycling of soil carbon, nitrogen, and other nutrients, yet we have a poor understanding of how soil microbiomes are shaped by their nutritional and physical environment. In this study, we investigated the successional dynamics of a soil microbiome during 21 weeks of enrichment on chitin and its monomer, N -acetylglucosamine. We examined succession of the soil communities in a physically heterogeneous soil matrix as well as a homogeneous liquid medium. The guiding hypothesis was that the initial species richness would influence the tendency for the selected consortia to stabilize and maintain a relatively constant community structure over time. We also hypothesized that long-term, substrate-driven growth would result in consortia with reduced species richness compared to the parent microbiome and that this process would be deterministic with relatively little variation between replicates. We found that the initial species richness does influence the long-term community stability in both liquid media and soil and that lower initial richness results in a more rapid convergence to stability. Despite use of the same soil inoculum and access to the same major substrate, the resulting community composition differed greatly in soil from that in liquid medium. Hence, distinct selective pressures in soils relative to homogenous liquid media exist and can control community succession dynamics. This difference is likely related to the fact that soil microbiomes are more likely to thrive, with fewer compositional changes, in a soil matrix than in liquid environments. The soil microbiome carries out important ecosystem functions, but interactions between soil microbial communities have been difficult to study due to the high microbial diversity and complexity of the soil habitat. In this study, we successfully obtained stable consortia with reduced complexity that contained species found in the original source soil. These consortia and the methods used to obtain them can be a valuable resource for exploration of specific mechanisms underlying soil microbial community ecology. The results of this study also provide new experimental context to better inform how soil microbial communities are shaped by new environments and how a combination of initial taxonomic structure and physical environment influences stability.

59 BASIC BIOLOGICAL SCIENCES↗

Substantial differences in soil viral community composition within and among four Northern California habitats

Abstract Viruses contribute to food web dynamics and nutrient cycles in diverse ecosystems, yet the biogeographical patterns that underlie these viral dynamics are poorly understood, particularly in soil. Here, we identified trends in soil viral community composition in relation to habitat, moisture content, and physical distance. We generated 30 soil viromes from four distinct habitats (wetlands, grasslands, woodlands, and chaparral) by selectively capturing virus-sized particles prior to DNA extraction, and we recovered 3432 unique viral ‘species’ (dsDNA vOTUs). Viral communities differed significantly by soil moisture content, with viral richness generally higher in wet compared to dry soil habitats. However, vOTUs were rarely shared between viromes, including replicates <10 m apart, suggesting that soil viruses may not disperse well and that future soil viral community sampling strategies may need to account for extreme community differences over small spatial scales. Of the 19% of vOTUs detected in more than one virome, 93% were from the same habitat and site, suggesting greater viral community similarity in closer proximity and under similar environmental conditions. Within-habitat differences indicate that extensive sampling would be required for rigorous cross-habitat comparisons, and results highlight emerging paradigms of high viral activity in wet soils and soil viral community spatial heterogeneity.

Durham, Devyn M.↗

Sustainability, Scalability, and Resiliency of the Town of Innisfil Mobility-on-Demand Experiment: Preliminary Results, Analyses, and Lessons Learned

In 2017, the town of Innisfil, Ontario, launched Innisfil Transit in partnership with Uber, a transportation network company, to provide a subsidized on-demand public mobility service as an alternative to investing in a new fixed-route bus service. The performance of Innisfil Transit is documented in a 2021 Ryerson University report by Sweet, Mitra, and Benaroya, which shows greater cost effectiveness of the mobility provided over the proposed bus alternative. This paper expands on those findings by assessing Innisfil Transit with respect to sustainability, scalability, and resiliency. First, we quantify the energy and emissions of this program relative to traditional transit and driving alone across varying powertrains. We then characterize a conservative first-order estimate of the percentage of US communities that fall within a similar spatial-demographic tier as Innisfil. Replicability also hinges on service cost and performance in comparison to average values for low-density transit in the US. Lastly, most transit agencies experienced a significant drop in demand (as much as 90%) with slowly rebounding ridership since the onset of the COVID-19 pandemic. The resiliency of the Innisfil program to the pressures induced by the pandemic is examined in comparison to other transit operations. The lessons learned across these three dimensions complement prior work to better understand the efficiency and sustainability of on-demand public mobility service for low-density communities like Innisfil.

ADVANCED PROPULSION SYSTEMS↗

Sustainability, Scalability and Resiliency of the Town of Innisfil Mobility-on-Demand Experiment: Preliminary Results, Analyses, and Lessons Learned: Preprint

In 2017, the Town of Innisfil, Ontario launched Innisfil Transit in partnership with Uber, a transportation network company, to provide a subsidized on-demand public mobility service as an alternative to investing in a new fixed-route bus service. The performance of Innisfil Transit is documented in a 2021 Ryerson University report which shows greater cost effectiveness of the mobility provided over the proposed bus alternative (Sweet, Mitra, and Benaroya 2021). This paper expands on those findings by assessing Innisfil Transit with respect to sustainability, scalability, and resiliency. First, we quantify the energy and emissions of this program relative to traditional transit and driving alone across varying powertrains. We then characterize a conservative first-order estimate of the percentage of US communities that fall within a similar spatial-demographic tier as Innisfil. Replicability also hinges on service cost and performance in comparison to average values for low-density transit in the US. Lastly, most transit agencies experienced a significant drop in demand (as much as 90%) with slowly rebounding ridership since the onset of the COVID-19 pandemic. The resiliency of the Innisfil program to the pressures induced by the pandemic is examined in comparison to other transit operations. The lessons learned across these three dimensions complement prior work to better understand the efficiency and sustainability of on-demand public mobility service for low-density communities like Innisfil.

ADVANCED PROPULSION SYSTEMS↗

Inverse deep learning methods and benchmarks for artificial electromagnetic material design

In this work we investigate the use of deep inverse models (DIMs) for designing artificial electromagnetic materials (AEMs) – such as metamaterials, photonic crystals, and plasmonics – to achieve some desired scattering properties (e.g., transmission or reflection spectrum). DIMs are deep neural networks (i.e., deep learning models) that are specially-designed to solve ill-posed inverse problems. There has recently been tremendous growth in the use of DIMs for solving AEM design problems however there has been little comparison of these approaches to examine their absolute and relative performance capabilities. In this work we compare eight state-of-the-art DIMs on three unique AEM design problems, including two models that are novel to the AEM community. Our results indicate that DIMs can rapidly produce accurate designs to achieve a custom desired scattering on all three problems. Although no single model always performs best, the Neural-Adjoint approach achieves the best overall performance across all problem settings. As a final contribution we show that not all AEM design problems are ill-posed, and in such cases a conventional deep neural network can perform better than DIMs. We recommend that a deep neural network is always employed as a simple baseline approach when addressing AEM design problems. Furthermore, we publish python code for our AEM simulators and our DIMs to enable easy replication of our results, and benchmarking of new DIMs by the AEM community.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

The Impact of Stand Age and Fertilization on the Soil Microbiome of Miscanthus × giganteus

Yield of the perennial grass Miscanthus × giganteus has shown an inconsistent and unpredictable response to nitrogen (N) fertilizer, yet fertilization underpins the crop’s environmental and economic sustainability. The interactions among soil microbial communities, N availability, and Miscanthus × giganteus and management may explain changes in plant productivity. In this study, soil samples from different stand ages of Miscanthus × giganteus in a replicated chronosequence field trial were used to investigate the effects of stand age and N fertilizer rates on microbial community structure. We hypothesized that there is a definable Miscanthus × giganteus soil microbiome and that this community varies significantly with stand age and fertilization. Our results showed that the main phyla in soil microbial communities, regardless of plant age, are similar but microbial community structures are significantly different. The variation in observed microbial communities generally decreases with older stand ages. The amount of N fertilizer applied also affected the microbial community structure associated with different aged Miscanthus × giganteus. Specifically, the relative abundance of Proteobacteria (Alphaproteobacteria and Gammaproteobacteria) and Acidobacteria (subgroup Gp1) increased shortly after fertilization and was more associated with younger Miscanthus × giganteus. Furthermore, our results show a significant relationship between bacterial α diversity and fertilization rates and that this response is also affected by stand age. Overall, our results emphasize linkages between microbial community structure, plant age, and fertilization in Miscanthus × giganteus.

59 BASIC BIOLOGICAL SCIENCES↗

Increased replication of dissimilatory nitrate-reducing bacteria leads to decreased anammox bioreactor performance

Background: Anaerobic ammonium oxidation (anammox) is a biological process employed to remove reactive nitrogen from wastewater. While a substantial body of literature describes the performance of anammox bioreactors under various operational conditions and perturbations, few studies have resolved the metabolic roles of their core microbial community members. Results: Here, we used metagenomics to study the microbial community of a laboratory-scale anammox bioreactor from inoculation, through a performance destabilization event, to robust steady-state performance. Metabolic analyses revealed that nutrient acquisition from the environment is selected for in the anammox community. Dissimilatory nitrate reduction to ammonium (DNRA) was the primary nitrogen removal pathway that competed with anammox. Increased replication of bacteria capable of DNRA led to the out-competition of anammox bacteria, and the loss of the bioreactor's nitrogen removal capacity. These bacteria were highly associated with the anammox bacterium and considered part of the core microbial community. Conclusions: Our findings highlight the importance of metabolic interdependencies related to nitrogen- and carbon-cycling within anammox bioreactors and the potentially detrimental effects of bacteria that are otherwise considered core microbial community members.

59 BASIC BIOLOGICAL SCIENCES↗

Streptavidin Affinity Grids for the Preparation of Biological Samples for Cryogenic Electron Microscopy (CRADA Final Report)

Currently, cryo-EM is the most powerful tool for structural analysis of large biomacromolecules, but there are still problems in that biomacromolecules are denatured at the air-interface and their orientations are limited in a specific orientation. We solved this problem at LBL by developing a streptavidin-coated EM grid. Use of such affinity-binding grids can minimize exposure to air-interface by keeping biomacromolecules bound to streptavidin. It is urgently needed to make such grids readily available to the cryo-EM community, but they are somewhat difficult to make. The goal of the CRADA with Hummingbird company was to mass-produce the streptavidin grids so that they would become readily available to the cryo-EM community. Our contribution to the CRADA was to first help Hummingbird to replicate the manual process that we use, and this was completed successfully.

59 BASIC BIOLOGICAL SCIENCES↗

City of Largo Clear Sky Assessment Process: Prioritizing Solar + Storage for Resilient Facilities & Communities

The Clear Sky Tampa Bay project was a 15-month collaborative research effort to support solar + storage deployment for community resilience in Florida. The Clear Sky Decision Support Toolkit is a collection of resources designed to support users in conducting solar + storage prioritization and feasibility screening assessments at critical facilities. The Tampa Bay Regional Planning Council worked with four local governments in the region to test and apply the Toolkit. This case study series describes how each partner government used the Toolkit and highlights key insights and lessons learned that other users could follow to replicate the process. The City of Largo used the Clear Sky Decision Support Toolkit to advance community solar + storage resilience goals by collaborating with staff across departments and disciplines and engaging with utility stakeholders. The City of Largo conducted an analysis of its Wastewater Treatment Facility (WWTF), which processes an average of 43 million gallons of wastewater daily. Water treatment operations are both critical and energy intensive, requiring sustained energy throughout the day.

14 SOLAR ENERGY↗

Propagation of viral genomes by replicating ammonia-oxidising archaea during soil nitrification

Ammonia-oxidising archaea (AOA) are a ubiquitous component of microbial communities and dominate the first stage of nitrification in some soils. While we are beginning to understand soil virus dynamics, we have no knowledge of the composition or activity of those infecting nitrifiers or their potential to influence processes. This study aimed to characterise viruses having infected autotrophic AOA in two nitrifying soils of contrasting pH by following transfer of assimilated CO 2 -derived 13 C from host to virus via DNA stable-isotope probing and metagenomic analysis. Incorporation of 13 C into low GC mol% AOA and virus genomes increased DNA buoyant density in CsCl gradients but resulted in co-migration with dominant non-enriched high GC mol% genomes, reducing sequencing depth and contig assembly. We therefore developed a hybrid approach where AOA and virus genomes were assembled from low buoyant density DNA with subsequent mapping of 13 C isotopically enriched high buoyant density DNA reads to identify activity of AOA. Metagenome-assembled genomes were different between the two soils and represented a broad diversity of active populations. Sixty-four AOA-infecting viral operational taxonomic units (vOTUs) were identified with no clear relatedness to previously characterised prokaryote viruses. These vOTUs were also distinct between soils, with 42% enriched in 13 C derived from hosts. The majority were predicted as capable of lysogeny and auxiliary metabolic genes included an AOA-specific multicopper oxidase suggesting infection may augment copper uptake essential for central metabolic functioning. These findings indicate virus infection of AOA may be a frequent process during nitrification with potential to influence host physiology and activity.

59 BASIC BIOLOGICAL SCIENCES↗