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DOE OSTI2020

Optimal Efficiency and Operational Cost Savings: A Framework for Automated Rooftop PV Placement

Residential energy consumers are charged based on a utility rate structure, such as net metering or feed-in tariff. To lower consumers' electricity bills, expensive batteries are deployed to reduce the electricity fed from the grid during peak hours. However, strategic photovoltaic (PV) panel placement enables the reduction of operational energy cost while considering the spatial feasibility and efficiency for hosting rooftop PV. In this paper, we present a framework to automatically identify the optimal location of rooftop PV panels on residential buildings. Our framework integrates multiple workflows, including energy and environmental simulation, parametric modeling, and optimization to identify the ideal location of PV panels to balance the demand and supply at a building and community scale. These workflows are linked using the Grasshopper plug-in for Rhinoceros CAD software. The framework includes two different workflows, each satisfying a target for optimal PV placement: (a) maximizing PV panel efficiency, where users aim to maximize energy generation, and (b) minimizing operational energy cost, where "best'' panels are selected considering utility rates for operational energy cost. Our framework is demonstrated in a residential community in Fort Collins, Colorado, to generate the optimal PV placement for each of the two aforementioned targets. Results from the two workflows are compared to illustrate the effect of PV location and orientation on solar energy production efficiency and operational energy cost. The developed workflows are introduced as tools within the Grasshopper plug-in to investigate the solar potential of rooftop PV panels while taking into account factors such as contextual shading, utility rate structures, and buildings' energy demand profiles.

30 DIRECT ENERGY CONVERSION↗

DOE OSTI2020

Simulation of Continental Shallow Cumulus Populations Using an Observation-Constrained Cloud-System Resolving Model

Continental shallow cumulus (ShCu) clouds observed on 30 August 2016 during the Holistic Interactions of Shallow Clouds, Aerosols, and Land-Ecosystems (HI-SCALE) field campaign are simulated by using an observation-constrained cloud-system resolving model. On this day, ShCu forms over Oklahoma and southern Kansas and some of these clouds transition to deeper, precipitating convection during the afternoon. We apply a four-dimensional ensemble-variational (4DEnVar) hybrid technique in the Community Gridpoint Statistical Interpolation (GSI) system to assimilate operational data sets and unique boundary layer measurements including a Raman lidar, radar wind profilers, radiosondes, and surface stations collected by the U.S. Department of Energy's (DOE) Atmospheric Radiation Measurement (ARM) Southern Great Plains (SGP) atmospheric observatory into the Weather Research and Forecasting (WRF) model to ascertain how improved environmental conditions can influence forecasts of ShCu populations and the transition to deeper convection. Independent observations from aircraft, satellite, as well as ARM's remote sensors are used to evaluate model performance in different aspects. Several model experiments are conducted to identify the impact of data assimilation (DA) on the prediction of clouds evolution. The analyses indicate that ShCu populations are more accurately reproduced after DA in terms of cloud initiation time and cloud base height, which can be attributed to an improved representation of the ambient meteorological conditions and the convective boundary layer. Extending the assimilation to 18 UTC (local noon) also improved the simulation of shallow-to-deep transitions of convective clouds.

54 ENVIRONMENTAL SCIENCES↗

NASA NTRS1984

Synoptic-scale dynamics with vertical velocity, part 1.8A

Radar measurements of all three of the atmospheric velocity components by the MST technique data from all the pioneering work of Woodman and Geillen (1974). The radar horizontal velocities have been compared with other standard measurements, such as radiosonde winds, in a number of studies and are now finding widespread acceptance within the meteorological community for research and operational forecasting purposes. Perhaps the single most interesting report recently is that the mesosphere-stratosphere-troposphere (MST) profiler winds are turning out to be one of the most useful pieces of data for predicting upslope snowfall in the cold season forecasting study of the PROFS Program (Reynolds, 1983). By contrast, the vertical velocities measured by MST radars have received relatively little attention, despite the facts that direct continuous measurement of vertical velocity is unique (i.e., it cannot be done with radiosondes) and that the vertical velocity is intimately linked with the dynamics of the atmosphere. Indeed, for many forecasting applications the vertical velocity is the single most important variable, yet it is usually inferred indirectly from other dynamical variables. The stratosphere-troposphere (ST) radars now available have the potential to change this situation. Some of the results from vertical velocity measurements which have direct application in synoptic scale dynamics.

Nastrom, G. D.↗

NASA NTRS2004

From Surface Chlorophyll a to Phytoplankton Community Composition in Oceanic Waters

The objective of the present study is to examine the potential of using the near-surface total chlorophyll a concentration (C(sub surf)), as it can be derived from ocean color observation, to infer the column-integrated and the vertical distribution of the phytoplanktonic biomass, both in a quantitative way and in a qualitative way (z.e., in terms of community structure). Within this context, a large HPLC (High Performance Liquid Chromatography) pigment database has been analyzed. It includes 2419 vertical pigment profiles, all sampled in Case-1 waters with various trophic states. The relationshps between C(sub surf) and the total chlorophyll alpha vertical distribution, as previously derived by Morel and Berthon, are fully confirmed, as the present results coincide with the previous ones. This agreement allows to go further, namely to examine the possibility of extracting relationships between C(sub surf) and the vertical composition of the algal assemblages. Thanks to the detailed pigment composition available from HPLC measurements, the contribution of three size classes (micro-, nano-, and pico-phytoplankton) to the local total chlorophyll a concentration can be assessed. Corroborating previous findings (e.g., large species dominate in eutrophc environments, whereas tiny phytoplankton prevail in oligotrophic zones), the results lead to a statistically based parameterization. The predictive skill of this parameterization is successfully tested on a separate data set. With such a tool, the vertical total chlorophyll a profiles associated with each size class can be inferred from the sole knowledge of C(sub surf). By combining this tool with satellite ocean color data, it becomes conceivable to quantify on a global scale the phytoplankton biomass associated with each of the three size classes.

Uitz, Julia↗

NASA NTRS2011

Evaluating the Impact of AIRS Observations on Regional Forecasts at the SPoRT Center

NASA Short-term Prediction Research and Transition (SPoRT) Center collaborates with operational partners of different sizes and operational goals to improve forecasts using targeted projects and data sets. Modeling and DA activities focus on demonstrating utility of NASA data sets and capabilities within operational systems. SPoRT has successfully assimilated the Atmospheric Infrared Sounder (AIRS) radiance and profile data. A collaborative project is underway with the Joint Center for Satellite Data Assimilation (JCSDA) to use AIRS profiles to better understand the impact of AIRS radiances assimilated within Gridpoint Statistical Interpolation (GSI) in hopes of engaging the operational DA community in a reassessment of assimilation methodologies to more effectively assimilate hyperspectral radiances.

Zavodsky, Bradley↗

NASA NTRS2022

Simulations of Firn Processes Over the Greenland and Antarctic Ice Sheets: 1980-2021

Conversion of altimetry-derived ice-sheet volume change to mass requires an understanding of the evolution of the combined ice and air content within the firn column. In the absence of suitable techniques to observe the changes to the firn column across the entirety of an ice sheet, the firn column processes are typically modeled. Here, we present new simulations of firn processes over the Greenland and Antarctic ice sheets (GrIS and AIS) using the Community Firn Model and atmospheric reanalysis variables for more than four decades. A data set of more than 250 measured depth–density profiles from both ice sheets provides the basis of the calibration of the dry-snow densification scheme. The resulting scheme results in a reduction in the rate of densification, relative to a commonly used semi-empirical model, through a decreased dependence on the accumulation rate, a proxy for overburden stress. The 1980–2020 modeled firn column runoff, when combined with atmospheric variables from MERRA-2, generates realistic mean integrated surface mass balance values for the Greenland (+390 Gt yr(exp −1)) and Antarctic (+2612 Gt yr(exp −1)) ice sheets when compared to published model-ensemble means. We find that seasonal volume changes associated with firn air content are on average approximately 2.5 times larger than those associated with mass fluxes from surface processes for the AIS and 1.5 times larger for the GrIS; however, when averaged over multiple years, ice and air-volume fluctuations within the firn column are of comparable magnitudes. Between 1996 and 2019, the Greenland Ice Sheet lost nearly 5% of its firn air content, indicating a reduction in the total meltwater retention capability. Nearly all (94 %) of the meltwater produced over the Antarctic Ice Sheet is retained within the firn column through infiltration and refreezing.

Greenland↗

DOE OSTI2023

How deep should we go to understand roots at the top of the world?

Harsh environmental conditions and the short summers of northern, high-latitude biomes impose unique constraints on the plants that live in the arctic tundra and the boreal forest. To escape the harsh aboveground environment, plants in these habitats often allocate a large portion of their biomass belowground to facilitate nutrient acquisition. In turn, the proximity of living plant roots to vast stores of sequestered soil carbon in these biomes means that shifts in rooting depth distribution and the size of the root–soil interface could significantly contribute to ongoing climate change. Indeed, plant ‘priming’ of rhizosphere decomposition via root exudation, particularly from shallowly distributed roots, can lead to losses of carbon from tundra soils. While we have a hard-won understanding of the distribution of plant communities across the arctic tundra and the boreal forest from direct field observations scaled to the landscape level using climate-informed mapping techniques (i.e. the Circumpolar Arctic Vegetation Map (CAVM); Walker et al. , 2005), these vegetation maps are only the tip of the iceberg (Iversen et al. , 2015). Root form and function remain hidden beneath the land surface. In an article recently published in New Phytologist , Blume-Werry et al. (2023, 10.1111/nph.18998) asked whether rooting depth distribution, and ensuing carbon emissions, could be inferred from commonly used vegetation mapping classifications across the pan-Arctic. An important question to guide our understanding, mapping, and prediction of belowground characteristics and ecosystem feedbacks at the top of the world. Unfortunately, they found that the answer was ‘not quite’. While rooting depth distribution varied demonstrably, in turn causing substantial changes in modeled carbon emissions via rhizosphere ‘priming’, variation across rooting depth profiles did not correspond with vegetation mapping classes. If we are unable to predict belowground rooting depth distributions across large spatial scales by leveraging aboveground vegetation community distributions, how then should belowground researchers proceed?

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS2019

Controlled Rest on the Flight Deck: Profile of Use, Challenges, and Best Practices

Managing fatigue in 24/7 operations is complex; fatigue and performance degradation can have health and safety consequences for the workers and can have economic and community consequences. We will present new findings on sleep, circadian misalignment, and performance in operational personnel. The presentations will shed light on how sleep and circadian misalignment affect workers' performance, safety, and health across multiple operations, and their application in fatigue risk management.As part of this symposium, I will discuss how pilots can use short naps (controlled rest) as a countermeasure to elevated sleepiness in-flight. My presentation will describe how and when controlled rest is used operationally and outline best practice guidelines for maximizing the benefits while managing the associated risks.

aviation↗

DOE OSTI2024

Metagenome-assembled genomes provide insight into the metabolic potential during early production of Hydraulic Fracturing Test Site 2 in the Delaware Basin

Demand for natural gas continues to climb in the United States, having reached a record monthly high of 104.9 billion cubic feet per day (Bcf/d) in November 2023. Hydraulic fracturing, a technique used to extract natural gas and oil from deep underground reservoirs, involves injecting large volumes of fluid, proppant, and chemical additives into shale units. This is followed by a “shut-in” period, during which the fracture fluid remains pressurized in the well for several weeks. The microbial processes that occur within the reservoir during this shut-in period are not well understood; yet, these reactions may significantly impact the structural integrity and overall recovery of oil and gas from the well. To shed light on this critical phase, we conducted an analysis of both pre-shut-in material alongside production fluid collected throughout the initial production phase at the Hydraulic Fracturing Test Site 2 (HFTS 2) located in the prolific Wolfcamp formation within the Permian Delaware Basin of west Texas, USA. Specifically, we aimed to assess the microbial ecology and functional potential of the microbial community during this crucial time frame. Prior analysis of 16S rRNA sequencing data through the first 35 days of production revealed a strong selection for a Clostridia species corresponding to a significant decrease in microbial diversity. Here, we performed a metagenomic analysis of produced water sampled on Day 33 of production. This analysis yielded three high-quality metagenome-assembled genomes (MAGs), one of which was a Clostridia draft genome closely related to the recently classified Petromonas tenebris. This draft genome likely represents the dominant Clostridia species observed in our 16S rRNA profile. Annotation of the MAGs revealed the presence of genes involved in critical metabolic processes, including thiosulfate reduction, mixed acid fermentation, and biofilm formation. These findings suggest that this microbial community has the potential to contribute to well souring, biocorrosion, and biofouling within the reservoir. Our research provides unique insights into the early stages of production in one of the most prolific unconventional plays in the United States, with important implications for well management and energy recovery.

natural gas↗

DOE OSTI2024

Pre-Bleaching Coral Microbiome Is Enriched in Beneficial Taxa and Functions

Coral reef health is tightly connected to the coral holobiont, which is the association between the coral animal and a diverse microbiome functioning as a unit. The coral holobiont depends on key services such as nitrogen and sulfur cycling mediated by the associated bacteria. However, these microbial services may be impaired in response to environmental changes, such as thermal stress. A perturbed microbiome may lead to coral bleaching and disease outbreaks, which have caused an unprecedented loss in coral cover worldwide, particularly correlated to a warming ocean. The response mechanisms of the coral holobiont under high temperatures are not completely understood, but the associated microbial community is a potential source of acquired heat-tolerance. Here we investigate the effects of increased temperature on the taxonomic and functional profiles of coral surface mucous layer (SML) microbiomes in relationship to coral–algal physiology. We used shotgun metagenomics in an experimental setting to understand the dynamics of microbial taxa and genes in the SML microbiome of the coral Pseudodiploria strigosa under heat treatment. The metagenomes of corals exposed to heat showed high similarity at the level of bacterial genera and functional genes related to nitrogen and sulfur metabolism and stress response. The coral SML microbiome responded to heat with an increase in the relative abundance of taxa with probiotic potential, and functional genes for nitrogen and sulfur acquisition. Coral–algal physiology significantly explained the variation in the microbiome at taxonomic and functional levels. These consistent and specific microbial taxa and gene functions that significantly increased in proportional abundance in corals exposed to heat are potentially beneficial to coral health and thermal resistance.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2022

Building the Next Generation Through 20 Years of Solar Decathlon: From Collegiate Prize to Multifaceted Clean Energy Workforce Program: Preprint

The U.S. Department of Energy Solar Decathlon, initiated in 2002 as a collegiate competition that challenges student teams to design and build high-performance, low-carbon buildings powered by renewables, has always been about developing students to be the next generation of buildings professionals. As we celebrate the 20th anniversary of the Solar Decathlon in 2022, this paper highlights its evolution from a singular prize to a multi-faceted platform preparing the broader buildings workforce to address the challenges of climate change. The original program challenged students to design and construct a residence. To improve the accessibility of the competition, the Design Challenge -- a 1-2 semester design-only competition-- was added to the Solar Decathlon platform. The Solar Decathlon has also scaled globally with events in Europe, the Middle East, China, Latin America and the Caribbean, Africa, and India. In 2021, the Solar Decathlon broadened its reach once again by introducing Solar Decathlon Pro, a zero-energy design practicum for professional architects and engineers. This paper utilizes participation data, surveys, alumni profiles, project submissions, and more to understand how changes to the program have expanded program reach to minority-serving institutions, environmental justice communities, and impacted the building industry's capacity to construct high performance and zero energy buildings. As one of the oldest federal government prizes, the Solar Decathlon offers a roadmap for dramatically increasing prize impact and demonstrates that it continues to push boundaries to tackle decarbonization and environmental justice.

build↗

NASA NTRSDate not supplied

Roadmap to Cooperative Operating Practices for Strategic Conflict Detection and Resolution in the Upper Class-E Traffic Management Concept

As the governing body of flight operations in the highly anticipated emergent area of Upper Class E airspace (60,000 ft and above), the Federal Aviation Administration (FAA) has recognized the potential for a possible extensible traffic management system for new entrants into this domain. Following the successes with Unmanned Aircraft Systems (UAS) Traffic Management (UTM) and Advanced / Urban Air Mobility (AAM / UAM) Traffic Management programs, FAA put forth an initial concept of operations for supporting the start of the Upper Class E Traffic Management (ETM) concept. Like UTM and AAM / UAM, ETM is envisioned to also be a community-based, industry driven cooperative management concept. However, tailoring it to be adaptable to the atmospheric communication, navigation, and surveillance deficits, as well as the diverse vehicle and mission profiles operating in the ETM environment will be the challenge. As such, the National Aeronautics and Space Administration (NASA) Ames Research Center has been investigating several technologies that will help enable industry in the development of this new type of cooperative operating environment. These technologies are being prototyped and will be tested in a collaborative evaluation of an initial ETM system in late 2023. The evaluation will concentrate on building out ETM system technologies that will inform the participants regarding operational intent sharing, strategic conflict detection, and the resultant deconfliction process. In addition to the technical aspects, key roles and responsibilities will need to be defined. This will be done through exploring community-agreed upon Cooperative Operating Practices (COPs) that include procedures and capabilities to aid in timely, strategic conflict identification and resolution to be developed during the evaluation. As an initial step to the evaluation, the ETM research team at NASA Ames solicited industry feedback on various aspects of ETM operations from subject matter experts. A virtual tabletop walkthrough session was held over a two-day period to follow a roadmap through the functional steps needed to build COPs, focusing on strategic conflict detection and resolution. Overall, the ETM tabletop provided insights into how the community wanted to instantiate the generation and sharing of operational intent, detect strategic conflicts and resolve those conflicts using a preliminary set of procedural community-agreed upon COPs.

Upper Class-E Traffic Management (ETM)↗

NASA NTRSDate not supplied

Roadmap to Cooperative Operating Practices for Strategic Conflict Detection and Resolution in the Upper Class E Traffic Management Concept

Upper Class E Traffic Management (ETM)↗

NASA NTRS2001

Ground-Based Icing Condition Remote Sensing System Definition

This report documents the NASA Glenn Research Center activities to assess and down select remote sensing technologies for the purpose of developing a system capable of measuring icing condition hazards aloft. The information generated by such a remote sensing system is intended for use by the entire aviation community, including flight crews. air traffic controllers. airline dispatchers, and aviation weather forecasters. The remote sensing system must be capable of remotely measuring temperature and liquid water content (LWC), and indicating the presence of super-cooled large droplets (SLD). Technologies examined include Profiling Microwave Radiometer, Dual-Band Radar, Multi-Band Radar, Ka-Band Radar. Polarized Ka-Band Radar, and Multiple Field of View (MFOV) Lidar. The assessment of these systems took place primarily during the Mt. Washington Icing Sensors Project (MWISP) in April 1999 and the Alliance Icing Research Study (AIRS) from November 1999 to February 2000. A discussion of the various sensing technologies is included. The result of the assessment is that no one sensing technology can satisfy all of the stated project goals. Therefore a proposed system includes radiometry and Ka-band radar. A multilevel approach is proposed to allow the future selection of the fielded system based upon required capability and available funding. The most basic level system would be the least capable and least expensive. The next level would increase capability and cost, and the highest level would be the most capable and most expensive to field. The Level 1 system would consist of a Profiling Microwave Radiometer. The Level 2 system would add a Ka-Band Radar. The Level 3 system would add polarization to the Ka-Band Radar. All levels of the system would utilize hardware that is already under development by the U.S. Government. However, to meet the needs of the aviation community, all levels of the system will require further development. In addition to the proposed system, it is also recommended that NASA continue to foster the development of Multi-Band Radar and airborne microwave radiometer technologies.

Reehorst, Andrew L.↗

DOE OSTI2025

Hybridization capture sequencing for Vibrio spp. and associated virulence factors

ABSTRACT Proliferation ofVibriospp. in aquatic ecosystems is associated with climate change and, concomitantly, increased incidence of vibriosis. They are autochthonous to aquatic environments globally, but traditional metagenomic methods for detecting and typing pathogenicVibriospp. are challenged by their presence in relatively low abundance and ability to persist in a viable but nonculturable state. In the study reported here, hybridization capture sequencing (HCS) was employed to profile low-abundanceVibriospp. in environmental samples. The HCS panel targeted a family of molecular chaperones (CPN60) specific to 69Vibriospp. and 162Vibrio-specific virulence factors. This approach was evaluated in parallel with traditional whole-community shotgun sequencing in a metagenomic analysis of water and oyster samples collected from the Chesapeake Bay. In addition,Vibrio parahaemolyticusandVibrio vulnificusstrains isolated from the samples were subjected to whole-genome sequencing to determine the genetic characteristics of pathogenicVibriospp. circulating in an aquatic environment. HCS, employed to determine the incidence and characterization of specificVibriospp., yielded significantly greater metagenomic insight, notably a variety of otherVibriospp., including detection ofVibrio cholerae,Vibrio fluvialis, andVibrio aestuarianus, in addition toVibrio parahaemolyticusandVibrio vulnificus, and also important virulence factors not detectable using traditional molecular methods. Thus, pathogenicVibriospp. in aquatic ecosystems may be far more common than currently understood. It is concluded that environmental surveillance should include HCS, a valuable tool for the detection and characterization of pathogenic agents in aquatic ecosystems, notably vibrios. IMPORTANCE The increasing prevalence of pathogenicVibriospp. in aquatic ecosystems, driven by climate change, is closely linked to a rise in cholera and vibriosis cases, emphasizing the need for improved environmental surveillance. Vibrios are naturally occurring in aquatic environments globally, but traditional metagenomic methods for detecting and typing pathogenicVibriospp. are challenged by their presence in relatively low abundance and ability to persist in a viable but nonculturable state. In the study reported here, hybridization capture sequencing was employed to profile low-abundanceVibriospp. in metagenomic samples, namely water and oysters collected from the Chesapeake Bay. This approach was evaluated in parallel with traditional whole-community shotgun sequencing and whole-genome sequencing ofVibrio parahaemolyticusandVibrio vulnificusstrains isolated from the samples. Results suggest pathogenicVibriospp. in aquatic ecosystems may be far more common than currently understood, when multiple methods are considered for environmental surveillance.

Microbiology↗

DOE OSTI2025

Hierarchical Resilience Planning for Networked Microgrids: A Case Study of Puerto Rico

Microgrids can be designed to enhance the energy resilience of communities and critical infrastructures, such as hospitals, data centers, and communication networks, which are vulnerable to frequent weather-related disruption. Coordinating multiple microgrids in a network can leverage the geographical diversity of load and generation resources while enabling resilient and cost-effective planning of the distribution system. Designing a networked microgrid is complex, involving intricate technical assessment, cost-benefit analysis, site-specific requirements, and the evaluation of existing resources. Therefore, this paper proposes a hierarchical resilience planning framework and performs an extensive techno-economic analysis for the design of a networked microgrid. Hierarchical resilience planning involves technology sizing at an individual community level to meet the critical load and satisfy resilience criteria, and resource optimization at networked microgrid level to provide a higher level of resilience and energy adequacy. A real-world case of Puerto Rico's cooperative microgrid “Microrred de la Montaña” is investigated considering localized electricity tariffs, site-specific demand profiles, solar generation, and existing hydro resources. Multiple optimization scenarios are developed based on the resiliency requirement to estimate the capacity of solar photovoltaic and battery energy storage (BES) to be installed at each substation. The results provide the optimal sizing for individual community and networked microgrid to withstand 1day and 3-day outages along with the criteria for critical load.

13 - HYDRO ENERGY↗

NASA NTRS2016

Radial Profile of the 3.5 kev Line Out to R200 in the Perseus Cluster

The recent discovery of the unidentified emission line at 3.5 keV in galaxies and clusters has attracted great interest from the community. As the origin of the line remains uncertain, we study the surface brightness distribution of the line in the Perseus cluster since that information can be used to identify its origin. We examine the flux distribution of the 3.5 keV line in the deep Suzaku observations of the Perseus cluster in detail. The 3.5 keV line is observed in three concentric annuli in the central observations, although the observations of the outskirts of the cluster did not reveal such a signal. We establish that these detections and the upper limits from the non-detections are consistent with a dark matter decay origin. However, absence of positive detection in the outskirts is also consistent with some unknown astrophysical origin of the line in the dense gas of the Perseus core, as well as with a dark matter origin with a steeper dependence on mass than the dark matter decay. We also comment on several recently published analyses of the 3.5 keV line.

dark matter â€“ elementary particles â€“ galaxies:↗

DOE OSTI2026

Cross-family and phage-specific gene requirements for Klebsiella infection revealed by scalable RB-TnSeq genetic screens.

Bacteriophages are being cataloged at an accelerating pace and are recognized as key players in nutrient and energy cycling across ecosystems. Yet the bacterial genetic determinants that govern phage-host specificity and infection success remain poorly understood, particularly in clinically and ecologically important genera such as Klebsiella where prior receptor characterization has been almost entirely limited to capsulated strains. Here we used a randomly barcoded, genome-wide, loss-of-function transposon mutant library (RB-TnSeq) of Klebsiella sp. M5al, a naturally acapsular, nitrogen-fixing rhizobacterium, to generate the first systematic, cross-family map of phage receptor gene dependencies in Klebsiella. Challenging the library against 25 double-stranded DNA phages spanning five families in 213 parallel assays, we identified 42 bacterial genes associated with phage infection, of which 15 had no prior association with phage infection in any bacterial system. Disruption of surface receptor biosynthesis genes conferred cross-resistance across multiple phage families, while intracellular gene disruptions had predominantly phage-specific effects. Clonal validation of eight genes confirmed LPS outer core biosynthesis genes as primary receptor determinants alongside additional host factors spanning outer membrane transport, cofactor biosynthesis, and two-component signaling. Comparative analysis across all 25 phages revealed that phage genus rather than family is the stronger predictor of host gene dependency profiles, a finding with direct implications for the functional annotation of uncharacterized phage isolates and rational phage cocktail design. Together, these findings provide a community resource for linking phage genomic diversity to functional host interaction space in this ecologically and clinically important genus.

Gittrich, Marissa R↗