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At least 325 records · Page 18

From viruses to protists: temperature response of the neglected components of microbial controls on peatland nutrient cycling

The response of plant-associated microbial communities to rising temperatures likely plays a key role in global Carbon (C) and Nitrogen (N) biogeochemical cycles. Mosses of the genus Sphagnum, in particular, sequester 25% of all terrestrial C as recalcitrant peat. However, their ability to sequester C is mediated by interactions with microbes that fix N and pass it to the moss to grow in otherwise oligotrophic systems. Two important biotic controls on this processes are likely to respond to rising temperatures –predation by protists and infections by viruses– but whether and how this might occur is not well understood. This proposal addressed these questions using a combination of genomics and mathematical modeling with both field data from whole-ecosystem warming experiments, and laboratory-based microcosm experiments. We have discovered 1) mechanistic links between cell traits and thermal performance in protists, which 2) enabled predictions from populations to ecosystems, 3) we have identified eco-phenotypic feedbacks that determine protist–bacterial predator–prey dynamics, 4) have documented concomitant compositional and trait shifts in protist communities with warming in the field, 5) revealed consistent interactive effects of temperature and nutrients on bacterial and protist communities; 6) characterized the Sphagnum virome, and 7) made modeling predictions on its implications for thermal responses of microbial communities in peatlands globally. This award also substantially strengthened U.S. scientific capacity and workforce development.

Gibert, Jean Philippe [Duke University]↗

Subsurface microbial communities as a tool for characterizing regional-scale groundwater flow

Subsurface microbial community distribution patterns are influenced by biogeochemical and groundwater fluxes and may inform hydraulic connections along groundwater-flow paths. This study examined the regional-scale microbial community of the Death Valley Regional Flow System and evaluated whether subsurface communities can be used to identify groundwater-flow paths between recharge and discharge areas. Samples were collected from 36 sites in three groundwater basins: Pahute Mesa–Oasis Valley (PMOV), Ash Meadows (AM), and Alkali Flat–Furnace Creek Ranch (AFFCR). Microbial diversity within and between communities varied by location, and communities were separated into two overall groups that affiliated with the AM and PMOV/AFFCR basins. Network analysis revealed patterns between clusters of common microbes that represented groundwaters with similar geochemical conditions and largely corroborated hydraulic connections between recharge and discharge areas. Null model analyses identified deterministic and stochastic ecological processes contributing to microbial community assemblages. Most communities were more different than expected and governed by dispersal limitation, geochemical differences, or undominating processes. However, certain communities from sites located within or near the Nevada National Security Site were more similar than expected and dominated by homogeneous dispersal or selection. Overall, the (dis)similarities between the microbial communities of DVRFS recharge and discharge areas supported previously documented hydraulic connections between: (1) Spring Mountains and Ash Meadows; (2) Frenchman and Yucca Flat and Amargosa Desert; and (3) Amargosa Desert and Death Valley. However, only a portion of the flow path between Pahute Mesa and Oasis Valley could be supported by microbial community analyses, likely due to well-associated artifacts in samples from the two Oasis Valley sites. This study demonstrates the utility of combining microbial data with hydrologic, geologic, and water-chemistry information to comprehensively characterize groundwater systems, highlighting both strengths and limitations of this approach.

54 ENVIRONMENTAL SCIENCES↗

Reducing uncertainty of high-latitude ecosystem models through identification of key parameters

Abstract Climate change is having significant impacts on Earth’s ecosystems and carbon budgets, and in the Arctic may drive a shift from an historic carbon sink to a source. Large uncertainties in terrestrial biosphere models (TBMs) used to forecast Arctic changes demonstrate the challenges of determining the timing and extent of this possible switch. This spread in model predictions can limit the ability of TBMs to guide management and policy decisions. One of the most influential sources of model uncertainty is model parameterization. Parameter uncertainty results in part from a mismatch between available data in databases and model needs. We identify that mismatch for three TBMs, DVM-DOS-TEM, SIPNET and ED2, and four databases with information on Arctic and boreal above- and belowground traits that may be applied to model parametrization. However, focusing solely on such data gaps can introduce biases towards simple models and ignores structural model uncertainty, another main source for model uncertainty. Therefore, we develop a causal loop diagram (CLD) of the Arctic and boreal ecosystem that includes unquantified, and thus unmodeled, processes. We map model parameters to processes in the CLD and assess parameter vulnerability via the internal network structure. One important substructure, feed forward loops (FFLs), describe processes that are linked both directly and indirectly. When the model parameters are data-informed, these indirect processes might be implicitly included in the model, but if not, they have the potential to introduce significant model uncertainty. We find that the parameters describing the impact of local temperature on microbial activity are associated with a particularly high number of FFLs but are not constrained well by existing data. By employing ecological models of varying complexity, databases, and network methods, we identify the key parameters responsible for limited model accuracy. They should be prioritized for future data sampling to reduce model uncertainty.

54 ENVIRONMENTAL SCIENCES↗

Linear two-pool models are insufficient to infer soil organic matter decomposition temperature sensitivity from incubations

Abstract Terrestrial carbon (C)-climate feedbacks depend strongly on how soil organic matter (SOM) decomposition responds to temperature. This dependency is often represented in land models by the parameter Q 10 , which quantifies the relative increase of microbial soil respiration per 10 °C temperature increase. Many studies have conducted paired laboratory soil incubations and inferred “active” and “slow” pool Q 10 values by fitting linear two-pool models to measured respiration time series. Using a recently published incubation study (Qin et al. in Sci Adv 5(7):eaau1218, 2019) as an example, here we first show that the very high parametric equifinality of the linear two-pool models may render such incubation-based Q 10 estimates unreliable. In particular, we show that, accompanied by the uncertain initial active pool size, the slow pool Q 10 can span a very wide range, including values as high as 100, although all parameter combinations are producing almost equally good model fit with respect to the observations. This result is robust whether or not interactions between the active and slow pools are considered (typically these interactions are not considered when interpreting incubation data, but are part of the predictive soil carbon models). This very large parametric equifinality in the context of interpreting incubation data is consistent with the poor temporal extrapolation capability of linear multi-pool models identified in recent studies. Next, using a microbe-explicit SOM model (RESOM), we show that the inferred two pools and their associated parameters (e.g., Q 10 ) could be artificial constructs and are therefore unreliable concepts for integration into predictive models. We finally discuss uncertainties in applying linear two-pool (or more generally multiple-pool) models to estimate SOM decomposition parameters such as temperature sensitivities from laboratory incubations. We also propose new observations and model structures that could enable better process understanding and more robust predictive capabilities of soil carbon dynamics.

54 ENVIRONMENTAL SCIENCES↗

Genomic reconstruction of fossil and living microorganisms in ancient Siberian permafrost

Abstract Background Total DNA (intracellular, iDNA and extracellular, eDNA) from ancient permafrost records the mixed genetic repository of the past and present microbial populations through geological time. Given the exceptional preservation of eDNA under perennial frozen conditions, typical metagenomic sequencing of total DNA precludes the discrimination between fossil and living microorganisms in ancient cryogenic environments. DNA repair protocols were combined with high throughput sequencing (HTS) of separate iDNA and eDNA fraction to reconstruct metagenome-assembled genomes (MAGs) from ancient microbial DNA entrapped in Siberian coastal permafrost. Results Despite the severe DNA damage in ancient permafrost, the coupling of DNA repair and HTS resulted in a total of 52 MAGs from sediments across a chronosequence (26–120 kyr). These MAGs were compared with those derived from the same samples but without utilizing DNA repair protocols. The MAGs from the youngest stratum showed minimal DNA damage and thus likely originated from viable, active microbial species. Many MAGs from the older and deeper sediment appear related to past aerobic microbial populations that had died upon freezing. MAGs from anaerobic lineages, including Asgard archaea, however exhibited minimal DNA damage and likely represent extant living microorganisms that have become adapted to the cryogenic and anoxic environments. The integration of aspartic acid racemization modeling and metaproteomics further constrained the metabolic status of the living microbial populations. Collectively, combining DNA repair protocols with HTS unveiled the adaptive strategies of microbes to long-term survivability in ancient permafrost. Conclusions Our results indicated that coupling of DNA repair protocols with simultaneous sequencing of iDNA and eDNA fractions enabled the assembly of MAGs from past and living microorganisms in ancient permafrost. The genomic reconstruction from the past and extant microbial populations expanded our understanding about the microbial successions and biogeochemical alterations from the past paleoenvironment to the present-day frozen state. Furthermore, we provided genomic insights into long-term survival mechanisms of microorganisms under cryogenic conditions through geological time. The combined strategies in this study can be extrapolated to examine other ancient non-permafrost environments and constrain the search for past and extant extraterrestrial life in permafrost and ice deposits on Mars.

54 ENVIRONMENTAL SCIENCES↗

Biomolecular budget of persistent, microbial-derived soil organic carbon: The importance of underexplored pools

The details of how soil microorganisms contribute to stable soil organic carbon pools are a pressing knowledge gap with direct implications for soil health and climate mitigation. It is now recognized that microbial necromass contributes substantially to the formation of stable soil carbon. However, the quantification of necromass in soils has largely been limited to model molecules such as aminosugar biomarkers. The abundance and chemical composition of other persistent microbial residues remain unresolved, particularly concerning how these pools may vary with microbial community structure, soil texture, and management practices. We use yearlong soil incubation experiments with an isotopic tracer to quantify the composition of persistent residues derived from microbial communities inhabiting sand or silt dominated soil with annual (corn) or perennial (switchgrass) monocultures. Persistent microbial residues were recovered in diverse soil biomolecular pools including metabolites, proteins, lipids, and mineral-associated organic matter (MAOM). The relative abundances of microbial contributions to necromass pools were consistent across cropping systems and soil textures. The greatest residue accumulation was not recovered in MAOM but in the light density fraction of soil debris that persisted after extraction by chemical fractionation using organic solvents. Necromass abundance was positively correlated with microbial biomass abundance and revealed a possible role of cell wall morphology in enhancing microbial carbon persistence; while gram-negative bacteria accounted for the greatest contribution to microbial-derived carbon by mass at one year, residues from gram-positive Actinobacteria and Firmicutes showed greater durability. Together these results offer a quantitative assessment of the relative importance of diverse molecular classes for generating durable soil carbon.

54 ENVIRONMENTAL SCIENCES↗

An Integrative Model for Soil Biogeochemistry and Methane Processes: I. Model Structure and Sensitivity Analysis

Abstract Environmental changes are anticipated to generate substantial impacts on carbon cycling in peatlands, affecting terrestrial‐climate feedbacks. Understanding how peatland methane (CH 4 ) fluxes respond to these changing environments is critical for predicting the magnitude of feedbacks from peatlands to global climate change. To improve predictions of CH 4 fluxes in response to changes such as elevated atmospheric CO 2 concentrations and warming, it is essential for Earth system models to include increased realism to simulate CH 4 processes in a more mechanistic way. To address this need, we incorporated a new microbial‐functional group‐based CH 4 module into the Energy Exascale Earth System land model (ELM) and tested it with multiple observational data sets at an ombrotrophic peatland bog in northern Minnesota. The model is able to simulate observed land surface CH 4 fluxes and fundamental mechanisms contributing to these throughout the soil profile. The model reproduced the observed vertical distributions of dissolved organic carbon and acetate concentrations. The seasonality of acetoclastic and hydrogenotrophic methanogenesis—two key processes for CH 4 production—and CH 4 concentration along the soil profile were accurately simulated. Meanwhile, the model estimated that plant‐mediated transport, diffusion, and ebullition contributed to ∼23.5%, 15.0%, and 61.5% of CH 4 transport, respectively. A parameter sensitivity analysis showed that CH 4 substrate and CH 4 production were the most critical mechanisms regulating temporal patterns of surface CH 4 fluxes both under ambient conditions and warming treatments. This knowledge will be used to improve Earth system model predictions of these high‐carbon ecosystems from plot to regional scales.

58 GEOSCIENCES↗

Numerical investigation of microbial quorum sensing under various flow conditions

Microorganisms efficiently coordinate phenotype expressions through a decision-making process known as quorum sensing (QS). We investigated QS amongst distinct, spatially distributed microbial aggregates under various flow conditions using a process-driven numerical model. Model simulations assess the conditions suitable for QS induction and quantify the importance of advective transport of signaling molecules. In addition, advection dilutes signaling molecules so that faster flow conditions require higher microbial densities, faster signal production rates, or higher sensitivities to signaling molecules to induce QS. However, autoinduction of signal production can substantially increase the transport distance of signaling molecules in both upstream and downstream directions. We present empirical approximations to the solutions of the advection–diffusion–reaction equation that describe the concentration profiles of signaling molecules for a wide range of flow and reaction rates. These empirical relationships, which predict the distribution of dissolved solutes along pore channels, allow to quantitatively estimate the effective communication distances amongst multiple microbial aggregates without further numerical simulations.

59 BASIC BIOLOGICAL SCIENCES↗

Coastal bacteria and protists assimilate viral carbon and nitrogen

Abstract Free viruses are the most abundant type of biological particles in the biosphere, but the lack of quantitative knowledge about their consumption by heterotrophic protists and bacterial degradation has hindered the inclusion of virovory in biogeochemical models. Using isotope-labeled viruses added to three independent microcosm experiments with natural microbial communities followed by isotope measurements with single-cell resolution and flow cytometry, we quantified the flux of viral C and N into virovorous protists and bacteria and compared the loss of viruses due to abiotic vs biotic factors. We found that some protists can obtain most of their C and N requirements from viral particles and that viral C and N get incorporated into bacterial biomass. We found that bacteria and protists were responsible for increasing the daily removal rate of viruses by 33% to 85%, respectively, compared to abiotic processes alone. Our laboratory incubation experiments showed that abiotic processes removed roughly 50% of the viruses within a week, and adding biotic processes led to a removal of 83% to 91%. Our data provide direct evidence for the transfer of viral C and N back into the microbial loop through protist grazing and bacterial breakdown, representing a globally significant flux that needs to be investigated further to better understand and predictably model the C and N cycles of the hydrosphere.

59 BASIC BIOLOGICAL SCIENCES↗

Genomic fingerprints of the world’s soil ecosystems

Despite the explosion of soil metagenomic data, we lack a synthesized understanding of patterns in the distribution and functions of soil microorganisms. These patterns are critical to predictions of soil microbiome responses to climate change and resulting feedbacks that regulate greenhouse gas release from soils. To address this gap, we assay 1,512 manually curated soil metagenomes using complementary annotation databases, read-based taxonomy, and machine learning to extract multidimensional genomic fingerprints of global soil microbiomes. Our objective is to uncover novel biogeographical patterns of soil microbiomes across environmental factors and ecological biomes with high molecular resolution. We reveal shifts in the potential for (i) microbial nutrient acquisition across pH gradients; (ii) stress-, transport-, and redox-based processes across changes in soil bulk density; and (iii) greenhouse gas emissions across biomes. We also use an unsupervised approach to reveal a collection of soils with distinct genomic signatures, characterized by coordinated changes in soil organic carbon, nitrogen, and cation exchange capacity and in bulk density and clay content that may ultimately reflect soil environments with high microbial activity. Genomic fingerprints for these soils highlight the importance of resource scavenging, plant-microbe interactions, fungi, and heterotrophic metabolisms. Across all analyses, we observed phylogenetic coherence in soil microbiomes—more closely related microorganisms tended to move congruently in response to soil factors. Collectively, the genomic fingerprints uncovered here present a basis for global patterns in the microbial mechanisms underlying soil biogeochemistry and help beget tractable microbial reaction networks for incorporation into process-based models of soil carbon and nutrient cycling.

59 BASIC BIOLOGICAL SCIENCES↗

Systems Analysis and Engineering of Biofuel Production in Chromochloris zofingiensis , an Emerging Model Green Alga

As a core component of a sustainable bioeconomy, microalgae have the potential to become a major source of biofuels and bioproducts. These photosynthetic microbes utilize solar energy, grow quickly, consume CO 2 , and can be cultivated on non-arable land. However, there are presently considerable practical limitations in the photosynthetic production of biofuels from microalgae, resulting in low productivity and high costs. Algae are a strikingly diverse group and understudied algae can reveal new opportunities for biofuels. Integrative systems biology and engineering of emerging model systems are needed to expand the possibilities of microbial production of biofuels and bioproducts. Our long-term goal is to design and engineer high-level production of biofuel precursors in microalgae.

09 BIOMASS FUELS↗

SetBERT: the deep learning platform for contextualized embeddings and explainable predictions from high-throughput sequencing

MOTIVATION: High-throughput sequencing (HTS) is a modern sequencing technology used to profile microbiomes by sequencing thousands of short genomic fragments from the microorganisms within a given sample. This technology presents a unique opportunity for artificial intelligence to comprehend the underlying functional relationships of microbial communities. However, due to the unstructured nature of HTS data, nearly all computational models are limited to processing DNA sequences individually. This limitation causes them to miss out on key interactions between microorganisms, significantly hindering our understanding of how these interactions influence the microbial communities as a whole. Furthermore, most computational methods rely on post-processing of samples which could inadvertently introduce unintentional protocol-specific bias. RESULTS: Addressing these concerns, we present SetBERT, a robust pre-training methodology for creating generalized deep learning models for processing HTS data to produce contextualized embeddings and be fine-tuned for downstream tasks with explainable predictions. By leveraging sequence interactions, we show that SetBERT significantly outperforms other models in taxonomic classification with genus-level classification accuracy of 95%. Furthermore, we demonstrate that SetBERT is able to accurately explain its predictions autonomously by confirming the biological-relevance of taxa identified by the model. AVAILABILITY AND IMPLEMENTATION: All source code is available at https://github.com/DLii-Research/setbert. SetBERT may be used through the q2-deepdna QIIME 2 plugin whose source code is available at https://github.com/DLii-Research/q2-deepdna.

Ludwig, David W↗

Geochemistry and Multiomics Data Differentiate Streams in Pennsylvania Based on Unconventional Oil and Gas Activity

Unconventional oil and gas (UOG) extraction is increasing exponentially around the world, as new technological advances have provided cost-effective methods to extract hard-to-reach hydrocarbons. While UOG has increased the energy output of some countries, past research indicates potential impacts in nearby stream ecosystems as measured by geochemical and microbial markers. Here, we utilized a robust data set that combines 16S rRNA gene amplicon sequencing (DNA), metatranscriptomics (RNA), geochemistry, and trace element analyses to establish the impact of UOG activity in 21 sites in northern Pennsylvania. These data were also used to design predictive machine learning models to determine the UOG impact on streams. We identified multiple biomarkers of UOG activity and contributors of antimicrobial resistance within the order Burkholderiales. Furthermore, we identified expressed antimicrobial resistance genes, land coverage, geochemistry, and specific microbes as strong predictors of UOG status. Of the predictive models constructed (n = 30), 15 had accuracies higher than expected by chance and area under the curve values above 0.70. The supervised random forest models with the highest accuracy were constructed with 16S rRNA gene profiles, metatranscriptomics active microbial composition, metatranscriptomics active antimicrobial resistance genes, land coverage, and geochemistry (n = 23). The models identified the most important features within those data sets for classifying UOG status. These findings identified specific shifts in gene presence and expression, as well as geochemical measures, that can be used to build robust models to identify impacts of UOG development.

16S rRNA↗

Subsurface H 2 Storage: A Williston Basin Commercial-Scale Resource Study

Poster for the 2024 NETL Resource Sustainability Project Review Meeting, Pittsburgh, Pennsylvania, April 2-4, 2024. This poster presents a commercial‑scale assessment of subsurface hydrogen storage potential in the North Dakota portion of the Williston Basin, evaluating saline formations, depleted oil and gas reservoirs, and salt formations. The study integrates laboratory characterization, reservoir simulation, and basinwide analysis to assess storage capacity, injectivity, recovery, and risks related to geochemical, microbial, and wellbore interactions. Results support the feasibility of large‑volume, secure hydrogen storage and provide a framework to guide future hydrogen commercialization and infrastructure development.

08 HYDROGEN↗

Microbially mediated mechanisms underlie soil carbon accrual by conservation agriculture under decade-long warming

Increasing soil organic carbon (SOC) in croplands by switching from conventional to conservation management may be hampered by stimulated microbial decomposition under warming. Here, we test the interactive effects of agricultural management and warming on SOC persistence and underlying microbial mechanisms in a decade-long controlled experiment on a wheat-maize cropping system. Warming increased SOC content and accelerated fungal community temporal turnover under conservation agriculture (no tillage, chopped crop residue), but not under conventional agriculture (annual tillage, crop residue removed). Microbial carbon use efficiency (CUE) and growth increased linearly over time, with stronger positive warming effects after 5 years under conservation agriculture. According to structural equation models, these increases arose from greater carbon inputs from the crops, which indirectly controlled microbial CUE via changes in fungal communities. As a result, fungal necromass increased from 28 to 53%, emerging as the strongest predictor of SOC content. Collectively, our results demonstrate how management and climatic factors can interact to alter microbial community composition, physiology and functions and, in turn, SOC formation and accrual in croplands.

59 BASIC BIOLOGICAL SCIENCES↗

Respiration data, microbial community assembly data, and FTICR-MS data associated with: “Disturbance Triggers Non-Linear Microbe-Environment Feedbacks. Sengupta et al., 2021, Biogeosciences”

This data package is associated with the manuscript “Disturbance Triggers Non-Linear Microbe-Environment Feedbacks, in revision in Biogeosciences (Sengupta et al. and 2021;https://bg.copernicus.org/preprints/bg-2021-51/). The study used hyporheic zone sediments as a model system to provide an integrated view of how disturbance modulates linkages among microbial ecology, biogeochemistry, and organic matter thermodynamics. Laboratory experiments exposed hyporheic sediment to varying wetting/drying dynamics. Data types include dissolved oxygen rates used to derive respiration rates, Fourier-transform ion cyclotron resonance mass spectrometry (FTICR-MS) data used to derive thermodynamic properties of organic matter, and microbial community assembly metrics derived from amplicon-sequence data of putatively active (cDNA) and whole community (gDNA). The outcomes of the study are condensed into a broadly applicable conceptual model linking external forcing, internal dynamics, and history. This data package is comprised of a file-level metadata (FLMD) csv, metadata csv, and seven folders that contain csv files, R scripts, xml files, and associated documentation: (1) Rates, (2) bNTI, and (3) FTICR, (4) Statistics_Analyses, (5) Raw OTU Beta dispersion Analysis, (6) bMNTD Randomizations, and (7) Data Dictionaries. The FLMD file has a description of each file included in the data package.

54 ENVIRONMENTAL SCIENCES↗

ENVnet provides a global molecular resource of dissolved organic matter

Dissolved organic matter (DOM) is an important component of Earth's carbon cycle and one of the planet's most chemically diverse pools, yet the molecular structures of its constituents remain largely unresolved. This limitation has hindered our ability to link DOM composition to microbial processes and ecosystem function. Here we present ENVnet, a global molecular repository built from tandem mass spectrometry data collected across 13 terrestrial and aquatic environment types, including 419 newly generated samples that expand publicly available DOM metabolomics data and cover previously underrepresented environments. By computationally deconvolving chimeric mass spectra, a longstanding challenge in environmental metabolomics, we recover high-quality fragmentation data for >22,000 distinct molecular features (defined by a specific precursor mass and fragmentation pattern). Using ENVnet, we uncover conserved and environment-specific molecular patterns in DOM composition and underlying biogeochemical processes. We also use molecular features encoded in ENVnet to train predictive models of DOM persistence, allowing molecular-level assessment of microbial turnover in independent systems.

54 ENVIRONMENTAL SCIENCES↗

An accurate and interpretable model for antimicrobial resistance in pathogenic Escherichia coli from livestock and companion animal species

Understanding the microbial genomic contributors to antimicrobial resistance (AMR) is essential for early detection of emerging AMR infections, a pressing global health threat in human and veterinary medicine. Here we used whole genome sequencing and antibiotic susceptibility test data from 980 disease causing Escherichia coli isolated from companion and farm animals to model AMR genotypes and phenotypes for 24 antibiotics. We determined the strength of genotype-to-phenotype relationships for 197 AMR genes with elastic net logistic regression. Model predictors were designed to evaluate different potential modes of AMR genotype translation into resistance phenotypes. Our results show a model that considers the presence of individual AMR genes and total number of AMR genes present from a set of genes known to confer resistance was able to accurately predict isolate resistance on average (mean F 1 score = 98.0%, SD = 2.3%, mean accuracy = 98.2%, SD = 2.7%). However, fitted models sometimes varied for antibiotics in the same class and for the same antibiotic across animal hosts, suggesting heterogeneity in the genetic determinants of AMR resistance. We conclude that an interpretable AMR prediction model can be used to accurately predict resistance phenotypes across multiple host species and reveal testable hypotheses about how the mechanism of resistance may vary across antibiotics within the same class and across animal hosts for the same antibiotic.

Chung, Henri C.↗