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At least 1,171 records · Page 65

Novel Modular Treatment System for Distributed Energy Recovery and Water Reclamation from Industrial Wastewaters

The overarching goal of this work was to accelerate the commercialization of a distributed, modular, agile treatment technology - the Modular Encapsulated Two-stage Anaerobic Biological (METAB) system - by advancing it to pilot-scale. Specifically, the METAB system was developed to treat high strength wastewater (5,000-35,000 mg/L chemical oxygen demand (COD)), produce hydrogen (H2) and methane (CH4), and achieve these outcomes without a membrane for retention of microorganisms.

42 ENGINEERING↗

Engineering Microbial Communities: Frontier Science for the Bioeconomy Workshop Series

In nature, biological systems are shaped by complex interactions of diverse microorganisms such as bacteria, archaea, fungi, and viruses living within communities called microbiomes (Berg et al. 2020; Prescott 2017). These collective interactions result in emergent community properties that can be leveraged for beneficial purposes such as bioenergy and biomolecule production. Given this potential and the immensity of microbial genomic diversity, the U.S. Department of Energy’s (DOE) Biological and Environmental Research (BER) program has long invested in research to better understand the biology of environmental microbes and microbiomes.

59 BASIC BIOLOGICAL SCIENCES↗

Characterization and Stimulation of Selenium Reducing Microbial Consortia from Real Flue Gas Desulfurization Wastewater Biological Treatment System

Selenium is released into the environment via anthropogenic activities such as coal combustion. The U.S. Environmental Protection Agency’s Effluent Limitation Guideline (ELG) Rule limits the release of selenium species into U.S. waterways. To meet the ELG limit for selenium, coal-fired power plants must treat the wastewater produced by the flue gas desulfurization (FGD) systems. Selenium removal is of interest because small changes of the concentrations of selenium in the environment can cause detrimental ecological consequences. Biological treatment is used to remediate this wastewater, but little is known about the individual species in the microbial consortium that reduce toxic water-soluble selenium oxyanions to insoluble elemental selenium. To optimize current biological treatment capabilities of this water, the selenium-reducing capabilities must be leveraged. In order to do so, the selenium-reducing bacteria (SeRB) in biological treatment systems must be characterized. This study will be the first to characterize the microbial consortia from a real FGD WW biological treatment system and provide insights on how to better achieve selenium removal via biological treatment. This work will provide insights into the microorganisms predominantly responsible for the reduction of selenium oxyanions in FGD wastewater biological treatment systems, which can be used to better optimize more efficient biological treatment strategies of this industrial waste.

geomicrobiology↗

Bioinorganic Chemistry of Nitrification: Structure and Function of Ammonia Monooxygenase (Final Technical Report)

The overarching goal of this project was to elucidate biochemical mechanisms that ubiquitous microorganisms use to enable primary metabolism based on nitrification, the multi-electron oxidation of ammonia (NH 3 ) to nitrite (NO 2 – ). This chemistry enables life to substitute carbon based fuel with nitrogen. Understanding the underlying chemistry was pursued to help inform the use of ammonia as an alternative fuel source compared to petroleum, capitalizing on the widespread infrastructure for production and distribution of this substance in the US. This project specifically focused on characterizing ammonia monooxygenase, an integral membrane copper protein that initiates nitrification via the aerobic oxidation of NH 3 to hydroxylamine (NH 2 OH).

09 BIOMASS FUELS↗

Engineering Anti-Microbial Peptide Production in Fungi and Bioprocess Development (CRADA 502)

Crop Protection, specifically controlling agricultural diseases and pests, is essential for crop production at every agricultural scale. Biologics hold enormous potential in this equation to decrease use of more toxic pesticides. Currently, the widespread use of biologics in crop protection is severely hampered by the lack of available tools for their mass production. To ensure biologics are regulated under EPA Biopesticide regulations, the following must be true: The biopesticide is a naturally occurring substance or structurally similar and functionally identical to a naturally occurring substance with a history of exposure to humans and the environment demonstrating minimal toxicity. Since this pertains to the marketed product it is also essential that fermentation hosts have a history of exposure and safe use by humans This research focuses on developing a novel platform for the production and delivery of the cghSAMP peptide using the filamentous fungus Aspergillus niger. A. niger is a well-established industrial microorganism, widely recognized for its strong protein secretion capabilities, high-yield production of enzymes, and classification as a generally recognized as safe (GRAS) organism. By genetically modifying A. niger to express the cghSAMP peptide transgene, we aim to leverage the fungus's robust fermentation system to produce a high volume of the antimicrobial agent. This engineered system represents a potential breakthrough in sustainable disease management, offering an environmentally friendly method for localized application or large-scale production of a biopesticide designed to mitigate the effects of citrus greening disease. The main objectives of this project were: 1. Developing a high-yield and cost-efficient bioprocess to produce an AMP in a current ABF host or readily onboarded hosts. 2. Scale up the process in stirred tank reactors and produce at least 100 grams of AMP at 1- 5g/L

60 APPLIED LIFE SCIENCES↗

Innovative Biomonitoring and Remediation of Heavy Metals Using Phytotechnologies at the Savannah River Site (SRS) Coal Combustion Product (CCP) Impoundment Sites

The Savannah River Site (SRS) contains legacy coal combustion product (CCP) impoundments that are impacted by elevated concentrations of heavy metals and radionuclides, posing long-term risks to soil health, ecosystem functioning, and environmental sustainability. Traditional environmental monitoring approaches rely primarily on chemical analyses to quantify contaminant concentrations but often provide limited information regarding biological responses or ecosystem recovery. This project addressed these limitations by integrating environmental chemistry, microbial ecology, artificial intelligence (AI), and bioremediation into a comprehensive framework for environmental diagnostics and restoration of contaminated soils. The overarching goal of this collaborative project between Florida A&M University (FAMU) and the University of Georgia's Savannah River Ecology Laboratory (SREL) was to develop innovative biomonitoring and remediation strategies for heavy metal-contaminated CCP impoundment sites at the Savannah River Site. Specifically, the project sought to (i) characterize heavy metal contamination, (ii) determine microbial responses to contamination, (iii) isolate indigenous heavy metal-resistant microorganisms for remediation applications, (iv) develop a microbial ecological health index using machine learning, and (v) optimize fungal-mediated bioremediation using artificial intelligence.

01 COAL, LIGNITE, AND PEAT↗

Final Technical Report for DE-SC0022206

This project developed foundational genetic, genomic, and epigenetic tools for anaerobic fungi (Neocallimastigomycota), a group of microorganisms with exceptional natural abilities to deconstruct lignocellulosic biomass. Efficient biomass deconstruction remains a major barrier to economical production of renewable fuels, chemicals, and materials from agricultural and forestry residues. The project sought to enable mechanistic studies and future engineering of anaerobic fungi by improving genomic resources, establishing methods for gene expression, and investigating epigenetic regulation of biomass-degrading pathways. Major accomplishments included generation of the first chromosome-scale genome assemblies for multiple anaerobic fungal species, providing publicly available genomic resources that support both engineering and fundamental biological research. The project established the first reproducible system for heterologous gene expression in anaerobic fungi and identified genomic features and mobile genetic elements that may support future development of stable transformation technologies. In parallel, the project demonstrated direct conversion of untreated lignocellulosic biomass into fuels and specialty chemicals through a fungal-yeast bioprocess and identified anaerobic fungal enzymes with utility for metabolic engineering. The research also revealed that epigenetic regulation plays an important role in controlling fungal gene expression and enzyme production, identifying potential strategies for enhancing biomass degradation. Collectively, this work established anaerobic fungi as a tractable emerging platform for bioenergy and biomanufacturing research, generated valuable public resources, trained the next generation of researchers, and advanced DOE-BER goals related to predictive biology, sustainable bioprocessing, and the circular bioeconomy.

Solomon, Kevin [University of Delaware] (ORCID:000↗

Predicting Metabolic Reaction Networks with Perturbation-Theory Machine Learning (PTML) Models

Background: Checking the connectivity (structure) of complex Metabolic Reaction Networks(MRNs) models proposed for new microorganisms with promising properties is an importantgoal for chemical biology. Objective: In principle, we can perform a hand-on checking (Manual Curation). However, this is achallenging task due to the high number of combinations of pairs of nodes (possible metabolic reactions). Results: The CPTML linear model obtained using the LDA algorithm is able to discriminate nodes(metabolites) with the correct assignation of reactions from incorrect nodes with values of accuracy,specificity, and sensitivity in the range of 85-100% in both training and external validation dataseries. Methods: In this work, we used Combinatorial Perturbation Theory and Machine Learning techniquesto seek a CPTML model for MRNs >40 organisms compiled by Barabasis’ group. First, wequantified the local structure of a very large set of nodes in each MRN using a new class of node indexcalled Markov linear indices fk. Next, we calculated CPT operators for 150000 combinationsof query and reference nodes of MRNs. Last, we used these CPT operators as inputs of differentML algorithms. Conclusion: Meanwhile, PTML models based on Bayesian network, J48-Decision Tree and RandomForest algorithms were identified as the three best non-linear models with accuracy greaterthan 97.5%. The present work opens the door to the study of MRNs of multiple organisms usingPTML models.

Pharmacology & Pharmacy↗

Micromundos biominerales en las algas rojas

Coralline algae (rhodophyta) populate vast pinkish colour regions of the coast. If you step on them in your bare feet, they might hurt you, because they are hard and sharp. Many organisms find shelter and develop within their tiny branches. Photosynthesis of coralline algae conducts the formation of carbonates that exhibit a fascinating architecture. The alga and its associated microorganisms (microbiota) participate in the formation of these minerals, that accumulate and cement the materials that ultimately shape beaches and coastal lines. Carbonates are susceptible to acid-base chemistry; thus, their structural stability and their dissolution depend on the pH of the surrounding environment. Therefore, these biominerals and the marine organisms that build them (such as algae, corals, mollusks or equinoderms) are vulnerable to ocean acidification. By trying to see beyond our eyesight, we were able to understand that algal branches hide an amazing structural strength, where its microstructure and chemistry play a major role. We found minerals with a vast structural and thermal stability in the algal body, named algal thallus. Currently, basic science explores coralline red and green algae as interesting models to understand carbon sequestration in stable structures. Therefore, this research might inspire the development of technologies to mitigate climate change.

59 BASIC BIOLOGICAL SCIENCES↗

Summary Data for paper titled: Influence of soil depth, irrigation, and plant genotype on the soil microbiome, metaphenome, and carbon chemistry

Climate change is causing an increase in drought in many soil ecosystems and a loss of soil organic carbon. Calcareous soils may partially mitigate these losses via carbon capture and storage. Here, we aimed to determine how irrigation-supplied soil moisture and perennial plants impact biotic and abiotic soil properties that underpin deep soil carbon chemistry in an unfertilized calcareous soil. Soil was sampled up to one meter in depth from irrigated and planted field treatments and analyzed using a suite of omics and chemical analyses. Carbon cycling processes in the surface soil were dominated by plant-microbe interactions that drive organic carbon cycling, whereas inorganic carbon chemistry dominated in deeper soil layers. Both irrigation and plant cover impacted organic and inorganic carbon pools in the soil profiles. This study reveals the complex interactions between water, plants, minerals, and microorganisms that govern organic and inorganic pools of soil carbon at different depths.

59 BASIC BIOLOGICAL SCIENCES↗

KS4A-Omics1.0_FspDS682

Soil fungi facilitate the translocation of inorganic nutrients from soil minerals to other microorganisms and plants. This ability is particularly advantageous in impoverished soils, because fungal mycelial networks can bridge otherwise spatially disconnected and inaccessible nutrient hotspots. However, the molecular mechanisms underlying fungal mineral weathering and transport through soil remains poorly understood. Here, we addressed this knowledge gap by directly visualizing nutrient acquisition and transport through fungal hyphae in a mineral doped soil micromodel using a multimodal imaging approach. Here, we observed how a representative of common saprotrophic soil fungi, Fusarium sp. DS 682, exhibited a mechanosensory response (thigmotropism) around obstacles and through pore spaces (~12 μm) in the presence of minerals.This study establishes the significance of fungal biology and nutrient translocation mechanisms in maintaining fungal growth under water and nutrient limitations in a soil-like microenvironment, using a high-throughput multi-omic analysis approach. Data package KS4A-Omics.1.0_FspDS682 (Publication: Fungal Mineral Weathering Mechanisms Revealed Through Direct Molecular Visualization) contents reported here are the first version (1.0) and contain pre- and post-processed data using high throughput data capture technologies for multi-omic analysis and integration, this data package contains raw and post-processed experimental data for X-Ray Absorption Near Edge Structure Spectroscopy (XANES/XRF), Optical Microscopy, Proteomics, Scanning Electron Microscope (SEM), Time-of-Flight Secondary Ion Mass Spectroscopy (ToF-SIMS), X-Ray Diffraction Spectroscopy (XRD) files, and X- Ray Photoelectron Spectroscopy (XPS) using EMSL capabilities. This data package DOI contains a comprehensive collection of high-throughput multi-omics data and process metadata catalog. Support files include additional data download contents “Read Me” with dataset descriptor information and data source method application ontologies (see data dictionary section). Reported data download content is structured for compliance with reported guidelines provided by community standard initiatives and publisher stakeholder policies supporting FAIR data principles.

47 OTHER INSTRUMENTATION↗

Data for Influence of soil depth, irrigation, and plant genotype on the soil microbiome, metaphenome, and carbon chemistry: Summary Data

Climate change is causing an increase in drought in many soil ecosystems and a loss of soil organic carbon. Calcareous soils may partially mitigate these losses via carbon capture and storage. Here, we aimed to determine how irrigation-supplied soil moisture and perennial plants impact biotic and abiotic soil properties that underpin deep soil carbon chemistry in an unfertilized calcareous soil. Soil was sampled up to one meter in depth from irrigated and planted field treatments and analyzed using a suite of omics and chemical analyses. Carbon cycling processes in the surface soil were dominated by plant-microbe interactions that drive organic carbon cycling, whereas inorganic carbon chemistry dominated in deeper soil layers. Both irrigation and plant cover impacted organic and inorganic carbon pools in the soil profiles. This study reveals the complex interactions between water, plants, minerals, and microorganisms that govern organic and inorganic pools of soil carbon at different depths.

Naasko, Katherine I↗

Data for Influence of soil depth, irrigation, and plant genotype on the soil microbiome, metaphenome, and carbon chemistry: Sequence Data

Climate change is causing an increase in drought in many soil ecosystems and a loss of soil organic carbon. Calcareous soils may partially mitigate these losses via carbon capture and storage. Here, we aimed to determine how irrigation-supplied soil moisture and perennial plants impact biotic and abiotic soil properties that underpin deep soil carbon chemistry in an unfertilized calcareous soil. Soil was sampled up to one meter in depth from irrigated and planted field treatments and analyzed using a suite of omics and chemical analyses. Carbon cycling processes in the surface soil were dominated by plant-microbe interactions that drive organic carbon cycling, whereas inorganic carbon chemistry dominated in deeper soil layers. Both irrigation and plant cover impacted organic and inorganic carbon pools in the soil profiles. This study reveals the complex interactions between water, plants, minerals, and microorganisms that govern organic and inorganic pools of soil carbon at different depths.

Naasko, Katherine I↗

Removal of primary nutrient degrading members severely reduces growth of soil microbial communities even when additional degraders are present

Understanding how microorganisms within a soil community interact to support collective respiration and growth remains challenging. Here we used a model substrate, chitin, and a Model Soil Consortium, MSC-2, to investigate how individual members of a microbial community contribute to decomposition and community growth. While MSC-2 can grow using chitin as the sole carbon source, we do not yet know how the growth kinetics or final biomass yields of MSC-2 vary when certain chitin degraders, or other important members, are absent. To characterize specific roles within this representative community, we carried out experiments leaving out members of MSC-2 and measuring biomass yields and CO2 production. We chose two members to iteratively leave out (referred to by genus name): Streptomyces, as it is predicted via gene expression analysis to be a major chitin degrader in the community, and Rhodococcus as it is predicted via species co-abundance analysis to interact with several other members. Our results showed that when MSC-2 lacked Streptomyces, growth and respiration of the community was severely reduced. Removal of either Streptomyces or Rhodococcus led to major changes in abundance for several other species, pointing to a comprehensive shifting of the microbial community when important members are removed as well as alterations in the metabolic profile, especially when Streptomyces was removed. These results show that when keystone, chitin degrading members are removed, other members, even those with the potential to degrade chitin, do not fill the same metabolic niche to promote community growth. In addition, highly connected members may be removed with similar or even increased levels of growth and respiration. Our findings are critical to a better understanding of soil microbiology, specifically in how communities maintain activity when biotic or abiotic factors lead to changes in biodiversity in soil systems.

McClure, Ryan S↗

WA-IsoC_MSC1.1.0

The soil microbiome is central to the cycling of carbon and other nutrients and to the promotion of plant growth. Despite its importance, analysis of the soil microbiome is difficult due to its sheer complexity, with thousands of interacting species. Here, we reduced this complexity by developing model soil microbial consortia that are simpler and more amenable to experimental analysis but still represent important microbial functions of the native soil ecosystem. Samples were collected from an arid grassland soil and microbial communities (consisting mainly of bacterial species) were enriched on agar plates containing chitin as the main carbon source. Chitin was chosen because it is an abundant carbon and nitrogen polymer in soil that often requires the coordinated action of several microorganisms for complete metabolic degradation. Several soil consortia were derived that had tractable richness (30-50 OTUs) with diverse phyla representative of the native soil, including Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria and Verrucomicrobia. The resulting consortia can be stored as glycerol or lyophilized stocks at -80° C and revived while retaining community composition, greatly increasing their use as tools for the research community at large. One of the consortia that was particularly stable was chosen as a model soil consortium (MSC-1) for further analysis. MSC-1 species interactions were studied using both pairwise co-cultivation in liquid media and during growth in soil under several perturbations. Co-abundance analyses highlighted interspecies interactions and helped to define keystone species, including Mycobacterium, Rhodococcus, and Rhizobiales taxa. These experiments demonstrate the success of an approach based on naturally enriching a community of interacting species that can be stored, revived, and shared. The knowledge gained from querying these communities and their interactions will enable better understanding of the soil microbiome and the roles these interactions play in this environment.

54 ENVIRONMENTAL SCIENCES↗

KBase Narrative - Comparing Proteomes of Cupriavidus basilensis 4G11 and Cupriavidus necator H16

Species of Cupriavidus are among the best bioplastic producing microorganisms known, though intracellular poly-hydroxybutyrate (PHB) is made exclusively during unbalanced growth (e.g. nitrogen limitation), a state characterized by poor growth and cellular stress. Much work has been done studying bioplastic production in the model/type strain C. necator H16. We use an RB-TnSeq library of the closely related C. basilensis 4G11 to assess for genes which decouple PHB production from growth state. In this narrative we compute genome-wide protein homology between these two organisms, and generate a synteny plot across the genomes as well.

Sander, Kyle↗

Osur_paper

Glyphosate is widely used as a herbicide, but recent studies begin to reveal its detrimental side effects on animals by targeting the shikimate pathway of associated gut microorganisms. However, its impact on nutritional endosymbionts in insects remains poorly understood. Here, we sequenced the tiny, shikimate pathway encoding symbiont genome of the sawtoothed grain beetle Oryzaephilus surinamensis. Decreased titers of the aromatic amino acid tyrosine in symbiont-depleted beetles underscore the symbionts’ ability to synthesize prephenate as the precursor for host tyrosine synthesis and its importance for cuticle sclerotization and melanization. Glyphosate exposure inhibited symbiont establishment during host development and abolished the mutualistic benefit on cuticle synthesis in adults, which could be partially rescued by dietary tyrosine supplementation. Furthermore, phylogenetic analyses indicate that the shikimate pathways of many nutritional endosymbionts likewise contain a glyphosate sensitive 5-enolpyruvylshikimate-3-phosphate synthase. These findings highlight the importance of symbiont-mediated tyrosine supplementation for cuticle biosynthesis in insects, but also paint an alarming scenario regarding the use of glyphosate in light of recent declines in insect populations.

59 BASIC BIOLOGICAL SCIENCES↗