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A compendium of bacterial and archaeal single-cell amplified genomes from oxygen deficient marine waters

Oxygen-deficient marine waters referred to as oxygen minimum zones (OMZs) or anoxic marine zones (AMZs) are common oceanographic features. They host both cosmopolitan and endemic microorganisms adapted to low oxygen conditions. Microbial metabolic interactions within OMZs and AMZs drive coupled biogeochemical cycles resulting in nitrogen loss and climate active trace gas production and consumption. Global warming is causing oxygen-deficient waters to expand and intensify. Therefore, studies focused on microbial communities inhabiting oxygen-deficient regions are necessary to both monitor and model the impacts of climate change on marine ecosystem functions and services. Here we present a compendium of 5,129 single-cell amplified genomes (SAGs) from marine environments encompassing representative OMZ and AMZ geochemical profiles. Of these, 3,570 SAGs have been sequenced to different levels of completion, providing a strain-resolved perspective on the genomic content and potential metabolic interactions within OMZ and AMZ microbiomes. Hierarchical clustering confirmed that samples from similar oxygen concentrations and geographic regions also had analogous taxonomic compositions, providing a coherent framework for comparative community analysis.

59 BASIC BIOLOGICAL SCIENCES↗

Comparative physiological and genomic characterization of a novel Nitrobacter vulgaris strain from a nitrate-contaminated subsurface

Nitrite-oxidizing bacteria (NOB) represent a crucial node in the global nitrogen cycle. By catalyzing the second step of nitrification—the oxidation of nitrite to nitrate to generate energy for growth—NOB activity controls the fate of nitrite (NO 2 - ) in aerobic environments. Despite thriving in diverse environments, including soils, freshwater, marine ecosystems, subsurface habitats, and water treatment systems, organisms capable of nitrite oxidation are confined to Nitrobacter, Nitrospira, Nitrospina, Nitrotoga, and a few other specific lineages. The genus Nitrobacter, recognized for its facultative heterotrophic metabolism, is often associated with high-nitrogen environments. Here, we report the physiological characterization of a novel strain, Nitrobacter vulgaris strain MLSD-S22, isolated from a nitrate- and heavy-metal-contaminated subsurface. Growth inhibition experiments revealed that strain MLSD-S22 and the N. vulgaris type strain Z exhibited similar sensitivities to nitrite and nitrate, with nitrite being the most inhibitory. Microrespirometry demonstrated that the two N. vulgaris strains and Nitrobacter winogradskyi Nb-255 possessed higher affinities for nitrite and oxygen than previously reported for Nitrobacter, suggesting potential to compete in low-substrate environments. Long-read DNA sequencing provided a complete genome for strain MLSD-S22, revealing two plasmids and an intact nitrous oxide (N 2 O) reduction operon—an unexpected feature for Nitrobacter. While N 2 O reduction activity was not observed under the tested conditions, this discovery raises questions about the contribution of Nitrobacter NOB to the N 2 O sink. These findings broaden the physiological and genomic diversity of Nitrobacter, offering new insights into their adaptation strategies and providing a framework for future evaluation of their potential roles in nitrogen loss.

Nitrobacter↗

DayCent_CABBI_energy_farm_sorghum_BNI_Ecosphere

This repository contains call source code and input files that are part of the DayCent_CABBI BNI model experiments described in the manuscript below. Hartman et al. 2022. In Silico Evaluation of Plant Nitrification Suppression Effects on Agroecosystem Nitrogen Loss. Ecosphere . In Review.

70101 Agricultural Land Management↗

Data from: Leaching losses of dissolved organic carbon and nitrogen from agricultural soils in the upper US Midwest

Leaching losses of dissolved organic carbon (DOC) and nitrogen (DON) from agricultural systems are important to water quality and carbon and nutrient balances but are rarely reported; the few available studies suggest linkages to litter production (DOC) and nitrogen fertilization (DON). In this study we examine the leaching of DOC, DON, NO3-, and NH4+ from no-till corn (maize) and perennial bioenergy crops (switchgrass, miscanthus, native grasses, restored prairie, and poplar) grown between 2009 and 2016 in a replicated field experiment in the upper Midwest U.S. Leaching was estimated from concentrations in soil water and modeled drainage (percolation) rates. DOC leaching rates (kg ha-1 yr-1) and volume-weighted mean concentrations (mg L-1) among cropping systems averaged 15.4 and 4.6, respectively; N fertilization had no effect and poplar lost the most DOC (21.8 and 6.9, respectively). DON leaching rates (kg ha-1 yr-1) and volume-weighted mean concentrations (mg L-1) under corn (the most heavily N-fertilized crop) averaged 4.5 and 1.0, respectively, which was higher than perennial grasses (mean: 1.5 and 0.5, respectively) and poplar (1.6 and 0.5, respectively). NO3- comprised the majority of total N leaching in all systems (59-92%). Average NO3- leaching (kg N ha-1 yr-1) under corn (35.3) was higher than perennial grasses (5.9) and poplar (7.2). NH4+ concentrations in soil water from all cropping systems were relatively low (<0.07 mg N L-1). Perennial crops leached more NO3- in the first few years after planting, and markedly less after. Among the fertilized crops, the leached N represented 14-38% of the added N over the study period; poplar lost the greatest proportion (38%) and corn was intermediate (23%). Requiring only one third or less of the N fertilization compared to corn, perennial bioenergy crops can substantially reduce N leaching and consequent movement into aquifers and surface waters.

09 BIOMASS FUELS↗

Soybean fallow and nitrification inhibitors: Strategies to reduce N 2 O emission intensities and N losses in Australian sugarcane cropping systems

The Australian sugar industry is facing mounting pressure to reduce the nitrogen (N) losses and nitrous oxide (N 2 O) emissions associated with N fertiliser use. Research has shown that N 2 O emissions from sugarcane (Saccharum officinarum L.) cropping systems can be reduced with the use of fertilisers coated with nitrification inhibitors, or by sowing legume crops in the fallows between sugarcane crop cycles. However, the efficacy of these two N management strategies across different climatic zones is still unclear, as results from field studies have been contradictory. The objectives of this study were therefore to use the DayCent model to assess the long-term effects of the two strategies (separately or in combination) on N 2 O emissions, N losses and yields. The model was parameterised using data from eight field experiments (39 treatments) conducted across four of the five main districts of the Australian sugarcane industry. A series of long-term scenarios embracing a range of N fertiliser rates (0- 160 kg N ha -1 applied as urea), climatic regions (Tropics and Subtropics), soil textures (fine- and coarse-textured soils) and N-loss risk scenarios (crop fertilised well before or close to the on-set of the wet season) were tested for both N management strategies. Simulations identified that the combined use of soybean (Glycine max L.) fallows and N fertiliser coated with the 3,4-dimethylpyrazole phosphate (DMPP) nitrification inhibitor was the most effective strategy to maintain or even increase current yields while significantly reducing N 2 O and cumulative N (mainly due to nitrate) losses. When used alone, the soybean fallow strategy on average reduced N losses over the entire crop cycle by 29 % compared with current industry best practice. However, this strategy showed a limited capacity to reduce N 2 O emissions over the entire cane crop cycle (median: -3% compared with current industry best practice), or even increased them in scenarios at high-risk of N losses in the Tropics. The use of urea coated with DMPP alone allowed current fertiliser N rates to be reduced by 30 % without any substantial yield penalty. In environments prone to high N losses, DMPP urea abated overall N losses and N 2 O emissions by 41 % and 30 %, respectively. This was the first study to use such an extensive dataset to parameterise a biogeochemical model for yields, N 2 O emissions and N losses in sugarcane cropping systems. Finally, the findings will be instrumental in guiding future research and policies designed to improve the profitability and environmental sustainability of this industry.

54 ENVIRONMENTAL SCIENCES↗

Arctic shrub expansion generates regional variation in litter decomposition by altering litter quality and the decomposition environment

Abstract The expansion of deciduous shrubs into the graminoid‐dominated arctic tundra is expected to alter litter decomposition by changing litter quality and local abiotic and biotic conditions. However, it remains unclear how shrub expansion will affect litter decomposition at regional scales, where macroclimate is expected to be the dominant regulator of decomposition. To determine the relative influence of macroclimate and local controls on regional patterns of litter mass loss and nitrogen release, we conducted two hierarchical litter decomposition experiments across spatial scales. We decomposed leaf and root litter from a prominent graminoid ( Eriophorum vaginatum ) and three genera of deciduous shrubs ( Betula , Alnus and Salix ) for 1 year within replicated plots at five sites spanning a 160 km latitudinal gradient in northern Alaska. Using Eriophorum litter as a substrate, we found that macroclimate was the primary regulator of mass loss but had opposing effects on leaf and root litter. As summer temperature increased along the latitudinal gradient (11.9 to 13.9°C), leaf litter mass loss increased by 20% whereas root litter mass loss decreased by 33%. Leaf nitrogen release also increased with summer temperature. Conversely, root nitrogen release was controlled by the vegetation type of the decomposition environment. Using different shrub litters as substrates, we found that litter quality and its interaction with soil microclimate and macroclimate controlled decomposition. Overall, shrub root litters decomposed faster than Eriophorum root litter, losing 53% more mass and 190% more nitrogen across all sites and decomposition environments. For leaf litter, however, patterns varied by litter genus, with Salix losing more mass and Betula and Alnus losing less mass than Eriophorum . Our findings demonstrate that shrub expansion in the Arctic can regulate leaf and root litter decomposition at the regional scale through its effects on local controls, primarily litter quality. Ongoing increases in shrub cover are likely to accelerate the turnover of root litter carbon and nitrogen pools in tundra ecosystems. Therefore, including shrub‐related processes in Earth system models will improve our ability to predict regional‐scale litter decomposition and its effects on carbon and nutrient cycling in a warming Arctic. Read the free Plain Language Summary for this article on the Journal blog.

Vozzo, Justin T. [Department of Natural Resources ↗

Grassland-to-cropland conversion increased soil, nutrient, and carbon losses in the US Midwest between 2008 and 2016

After decades of declining cropland area, the United States (US) experienced a reversal in land use/land cover change in recent years, with substantial grassland conversion to cropland in the US Midwest. Although previous studies estimated soil carbon (C) loss due to cropland expansion, other important environmental indicators, such as soil erosion and nutrient loss, remain largely unquantified. In this study, we simulated the environmental impacts from the conversion of grassland to corn and soybeans for 12 US Midwestern states using the EPIC (Environmental Policy Integrated Climate) model. Between 2008 and 2016, over 2 Mha of grassland were converted to crop production in these states, with much less cropland concomitantly abandoned or retired from production. The net grassland-cropland conversion increased annual soil erosion by 7.9%, nitrogen (N) loss by 3.7%, and soil organic carbon loss by 5.6% relative to that of existing cropland, despite an associated increase in cropland area of only 2.5%. Notably, the above estimates represent the scenario of converting unmanaged grassland to tilled corn and soybeans, and impacts varied depending upon crop type and tillage regime. Corn and soybeans are dominant biofuel feedstocks, yet the grassland conversion and subsequent environmental impacts simulated in this study are likely not attributable solely to biofuel-driven land use change since other factors also contribute to corn and soybean prices and land use decisions. Nevertheless, our results suggest grassland conversion in the Upper Midwest has resulted in substantial degradation of soil quality, with implications for air and water quality as well. Additional conservation measures are likely necessary to counterbalance the impacts, particularly in areas with high rates of grassland conversion (e.g. the Dakotas, southern Iowa).

54 ENVIRONMENTAL SCIENCES↗

Effects of Media Nutrient Variation on Microalgae Productivity and Economics During Semi-Continuous Cultivation

The development of large-scale microalgae growth for biofuel production is currently limited by the cost of biomass production. However, new approaches to infrastructure and cultivation practices are bringing the field closer to realization. Macronutrients in the cultivation media contribute significant costs, especially since their concentrations have not been optimized for specific strains and conditions. Environmental photobioreactors (ePBRs) were used to simulate cultivation under outdoor conditions, during which the nitrogen and phosphorus levels in the media were varied. The growth of two potential biofuel production strains, Picochlorum celeri and Tetraselmis striata, with varying nutrient inputs during summer and winter scripts, respectively, was studied. This study demonstrated that nitrogen and phosphorus in f/2 media could be reduced by more than 60% from the standard formulation, while maintaining growth rates in a semi-continuous harvesting approach. Experiments comparing the standard and reduced nutrient input concentrations were also conducted for both species in 820 L outdoor raceway ponds, in Mesa, AZ. P. celeri grown in these ponds in October had a growth rate of 10.6 ± 0.7 g/m 2 /day and 10.6 ± 0.3 g/m 2 /day for the standard and low-nutrient P. celeri ponds, respectively. T. striata grown in April–May had a growth rate of 16.6 ± 1.4 g/m 2 /day for the standard nutrient input ponds and 17.4 ± 1.1 g/m 2 /day for the low-nutrient input ponds, and in October 14.5 ± 0.6 g/m 2 /day for standard nutrient ponds and 14.4 ± 0.6 g/m 2 /day for low-nutrient ponds. These outdoor data therefore confirmed the indoor ePBR data. Techno-economic analysis shows that, if high growth rates can be attained at lower nutrient concentrations, a reduction of at least 60% in nutrient costs can be achieved. Such results highlight the importance of managing macronutrient media inputs, as these have a considerable contribution to biomass production costs in large-scale facilities. The analysis also points to the importance of maintaining high spent medium recycling rates in an industrial deployment, so as to minimize the losses of nitrogen and phosphorus compounds.

09 BIOMASS FUELS↗

Utilizing Novel Field and Data Exploration Methods to Explore Hot Moments in High-Frequency Soil Nitrous Oxide Emissions Data: Opportunities and Challenges

Soil nitrous oxide (N 2 O) emissions are an important driver of climate change and are a major mechanism of labile nitrogen (N) loss from terrestrial ecosystems. Evidence increasingly suggests that locations on the landscape that experience biogeochemical fluxes disproportionate to the surrounding matrix (hot spots) and time periods that show disproportionately high fluxes relative to the background (hot moments) strongly influence landscape-scale soil N 2 O emissions. However, substantial uncertainties remain regarding how to measure and model where and when these extreme soil N 2 O fluxes occur. High-frequency datasets of soil N 2 O fluxes are newly possible due to advancements in field-ready instrumentation that uses cavity ring-down spectroscopy (CRDS). Here, we outline the opportunities and challenges that are provided by the deployment of this field-based instrumentation and the collection of high-frequency soil N 2 O flux datasets. While there are substantial challenges associated with automated CRDS systems, there are also opportunities to utilize these near-continuous data to constrain our understanding of dynamics of the terrestrial N cycle across space and time. Finally, we propose future research directions exploring the influence of hot moments of N 2 O emissions on the N cycle, particularly considering the gaps surrounding how global change forces are likely to alter N dynamics in the future.

54 ENVIRONMENTAL SCIENCES↗

Water Quality Effects of Economically Viable Land Use Change in the Mississippi River Basin under the Renewable Fuel Standard

Demand for biofuel production driven by the Renewable Fuel Standard (RFS2) has coincided with increased land in corn production and increasing nitrogen (N) loss to the Gulf of Mexico. Diversifying cropland with perennial energy crops (miscanthus and switchgrass) may reduce N loss and improve water quality. However, the extent of these benefits depends on the mix of biomass feedstocks (corn stover, perennials) incentivized by the RFS2 and the extent to which energy crops displace N-intensive row crops. We developed an integrated economic-biophysical model to quantify the water quality impacts of three potential policy scenarios that provided corn ethanol at levels before the RFS2 (RFS1 baseline); 15 billion gallons of corn ethanol (corn ethanol only); or 16 billion gallons of cellulosic ethanol in addition to corn ethanol (corn + cellulosic ethanol). Our results showed that economically optimal locations for perennial energy crop production were distributed across idle cropland with lower intrinsic N loss than active cropland. We found stover removal incentivized by the RFS2 offset N loss benefits of perennial energy crops. Furthermore, this finding suggests that targeted incentives for N loss reduction are needed to supplement the RFS2 to induce displacement of N-intensive row crops with energy crops to reduce N losses.

54 ENVIRONMENTAL SCIENCES↗

Printed Potentiometric Ammonium Sensors for Agriculture Applications

Ammonium (NH 4 + ) concentration is critical to both nutrient availability and nitrogen (N) loss in soil ecosystems but can be highly variable across spatial and temporal scales. For this reason, effectively informing agricultural practices such as fertilizer management and understanding of mechanisms of soil N loss require sensor technologies to monitor ammonium concentrations in real time. Our work investigates the performance of fully printed ammonium ion-selective sensors used in diverse soil environments. Ammonium sensors consisting of a printed ammonium ion-selective electrode and a printed Ag/AgCl reference were fabricated and characterized in aqueous solutions and three different soil types (sand, peat, and clay) under the range of ion concentrations likely to be present in soil (0.01–100 mM). The response of ammonium sensors was further evaluated under variable gravimetric moisture content in the soil to reflect their reliability under field conditions. Ammonium sensors demonstrated a sensitivity of 53.6 ± 5.1 mV/decade when tested in aqueous solution, and a sensitivity of 55.7 ± 11 mV/dec, 57.5 ± 4.1 mV/dec, and 43.7 ± 4 mV/dec was measured in sand, clay, and peat soils, respectively.

60 APPLIED LIFE SCIENCES↗

Extrapolation Domains for Aggregating Environmental Outcomes from Local to Regional Levels

Billions of dollars are invested every year to run field experiments to quantify the response of crops to new technologies and associated environmental outcomes. Given the diversity of environments where crop production occurs, conducting these research studies without a robust framework for research site selection and upscaling results to larger spatial scales is inefficient. In this project, researchers from the University of Nebraska-Lincoln used their unique Technology Extrapolation Domains framework (TED) as a tool to guide the selection of experimental sites and as basis for aggregating and validating environmental outcomes from local to regional levels. In the present project, we first used the TED framework to evaluate the current distribution and area coverage of the current SMARTFARM sites. The purpose was to showcase how it is possible to use the TED framework to guide site selection and extrapolate results over space. Additionally, the original TED framework was expanded to account for other factors influencing environmental outcomes by inclusion of three additional variables: soil organic matter, soil texture, and topographic wetness index. The resulting expanded framework (TED-E) was validated using nitrogen (N) losses from corn in the United States as a case study. To do so, we used N balance as a proxy to N losses and we evaluated the capacity of the framework to explain variation in N balance across fields and across countries. We found that the TED-E has substantially higher predictive power than the original TED framework to explain spatial variation in N balance. However, improvements in predictive power with the TED-E tool come at the expense of a higher number of TEDs needed to achieve a given crop area coverage compared with the original TED framework. We conclude that the new TED-E framework can help aggregate and extrapolate environmental outcomes from research sites to regional levels and improve the visualization of their spatial patterns across the United States. An online version of the framework is available at: https://www.toolted.org/

54 ENVIRONMENTAL SCIENCES↗

Nitrogen fixation rates in the Guinea Dome and the equatorial upwelling regions in the Atlantic Ocean

Abstract Biological nitrogen fixation is a key process balancing the loss of combined nitrogen in the marine nitrogen cycle. Its relevance in upwelling or high nutrient regions is still unclear, with the few available studies in these regions of the ocean reporting rates that vary widely from below detection limit to > 100 nmol N L −1 d −1 . In the eastern tropical Atlantic Ocean, two open ocean upwelling systems are active in boreal summer. One is the seasonal equatorial upwelling, where the residual phosphorus associated with aged upwelled waters is suggested to enhance nitrogen fixation in this season. The other is the Guinea Dome, a thermal upwelling dome. We conducted two surveys along 23° W across the Guinea Dome and the Equator from 15° N to 5° S in September 2015 and August–September 2016 with high latitudinal resolution (20–60 nm between stations). The abundance of Trichodesmium colonies was characterized by an Underwater Vision Profiler 5 and the total biological nitrogen fixation in the euphotic layer was measured using the 15 N 2 technique. The highest abundances of Trichodesmium colonies were found in the area of the Guinea Dome (9°–15° N) with a maximum of 3 colonies L −1 near the surface. By contrast, colonies were almost absent in the Equatorial band between 2° N and 5° S. The highest nitrogen fixation rate was measured at the northern edge of the Guinea Dome in 2016 (ca. 31 nmol N L −1 d −1 ). In this region, where diazotrophs thrived on a sufficient supply of both phosphorus and iron, a patchy distribution was unveiled by our increased spatial resolution scheme. In the Equatorial band, rates were considerably lower, ranging from below detection limit to ca. 4 nmol N L −1 d −1 , with a clear difference in magnitude between 2015 (rates close to zero) and 2016 (average rates around 2 nmol N L −1 d −1 ). This difference seemed triggered by a contrasting supply of phosphorus between years. Our study stresses the importance of surveys with sampling at fine-scale spatial resolution, and shows unexpected high variability in the rates of nitrogen fixation in the Guinea Dome, a region where diazotrophy is a significant process supplying new nitrogen into the euphotic layer.

Fernández-Carrera, Ana (ORCID:0000000183470996)↗

Minimizing the impacts of the ammonia economy on the nitrogen cycle and climate

Ammonia (NH 3 ) is an attractive low-carbon fuel and hydrogen carrier. However, losses and inefficiencies across the value chain could result in reactive nitrogen emissions (NH 3 , NO x , and N 2 O), negatively impacting air quality, the environment, human health, and climate. A relatively robust ammonia economy (30 EJ/y) could perturb the global nitrogen cycle by up to 65 Mt/y with a 5% nitrogen loss rate, equivalent to 50% of the current global perturbation caused by fertilizers. Moreover, the emission rate of nitrous oxide (N 2 O), a potent greenhouse gas and ozone-depleting molecule, determines whether ammonia combustion has a greenhouse footprint comparable to renewable energy sources or higher than coal (100 to 1,400 gCO 2 e/kWh). The success of the ammonia economy hence hinges on adopting optimal practices and technologies that minimize reactive nitrogen emissions. We discuss how this constraint should be included in the ongoing broad engineering research to reduce environmental concerns and prevent the lock-in of high-leakage practices.

54 ENVIRONMENTAL SCIENCES↗

Temperature Mapping analysis of Nitrogen-doped Niobium Radio Frequency Cavity

Nitrogen-doping of niobium superconducting radio-frequency (SRF) cavity has been shown to significantly improve the quality factors as compared to the standard treated niobium cavities. However, the nitrogen-doped cavities generally quench at lower accelerating gradients. To understand the possible sources of rf losses in nitrogen-doped cavities that could reduce the quench field, we used 576 thermometer sensors to map the temperature distribution of the outer cavity surface while increasing the accelerating field. Our measurements have revealed a strong effect of the cavity cooldown rate on the intensity of hot spots and their spatial distribution, indicating a significant contribution from trapped vortices on the RF losses.

Makita, Junki↗

Soil net nitrogen mineralization and leaching under Miscanthus × giganteus and Zea mays

Abstract The winter fallow period common in annual cropping systems leaves soils vulnerable to erosion and nutrient loss, especially to nitrogen (N) leaching. This vulnerability can be mitigated with perennial crops that have living roots in the ground year‐round. The mechanisms, magnitude, and consistency with which perennial crops retain N are not clear. We used an experiment to test whether a perennial crop, miscanthus ( Miscanthus × giganteus Greef et Deu.), would leach less N than continuous maize ( Zea mays L.) and how soil net N mineralization (N min ) may explain observed leaching under varied environment and management conditions. The experiment included three crossed factors: (1) cropping system (maize, juvenile miscanthus = 1–2 years old, mature miscanthus = 3–4 years old); (2) N fertilization (0 and 224 kg N ha −1 ); and (3) environment (four site‐years at two locations in Iowa, USA, that differed in climate and soil fertility). We measured N cycling dynamics, including: inorganic soil N (ammonium + nitrate), in situ N min , N leaching, crop N uptake, and calculated system N use efficiency. There were many complex interactions among factors. On average, cumulative N min under juvenile miscanthus was 111% greater than maize, but as miscanthus matured, there was no difference in N min between the perennial crop and maize. There was no difference in N leaching between juvenile miscanthus and maize, but mature miscanthus decreased N leaching by 42% and 88% compared to maize (with and without N fertilization, respectively). Across all treatments, there was no relationship between N min and N leaching, suggesting other mechanisms are regulating N leaching. Overall, mature miscanthus shows promise as a tool to reduce N losses in areas dominated by annual row‐crops.

54 ENVIRONMENTAL SCIENCES↗