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At least 361 records · Page 20

Detection and Attribution of the Recovery of Polar Ozone

The Antarctic ozone hole develops each year and culminates by early spring (late September - early October). The severity of the hole has been assessed from satellites using the minimum total ozone value from the October monthly mean (depth of the hole), calculating the average area coverage during this September-October period, and by estimating ozone mass deficit. Profile information shows that ozone is completely destroyed in the 14-2 1 km layer by early October. Ozone is mainly destroyed by halogen (chlorine and bromine) catalytic cycles, and these losses are modulated by temperature variations. Because atmospheric halogen levels are responding to international agreements that limit or phase out production, the amount of halogens in the stratosphere should decrease over the next few decades. Both models and projections of ozone depleting substances (ODSs) into the 21St century reveal that polar ozone levels should recover in the 2060- 2070 period. In this talk, we will review current projections of polar ozone recovery. Using models and ODs projections, we explore both the past, near future (2008-2025), and far future (> 2025) levels of polar ozone. Finally, we will discuss various factors that complicate recovery such as greenhouse gas changes (e.g., cooling in the upper stratosphere) and the acceleration of the Brewer-Dobson circulation.

Newman, Paul A.↗

Model for Simulating a Spiral Software-Development Process

A discrete-event simulation model, and a computer program that implements the model, have been developed as means of analyzing a spiral software-development process. This model can be tailored to specific development environments for use by software project managers in making quantitative cases for deciding among different software-development processes, courses of action, and cost estimates. A spiral process can be contrasted with a waterfall process, which is a traditional process that consists of a sequence of activities that include analysis of requirements, design, coding, testing, and support. A spiral process is an iterative process that can be regarded as a repeating modified waterfall process. Each iteration includes assessment of risk, analysis of requirements, design, coding, testing, delivery, and evaluation. A key difference between a spiral and a waterfall process is that a spiral process can accommodate changes in requirements at each iteration, whereas in a waterfall process, requirements are considered to be fixed from the beginning and, therefore, a waterfall process is not flexible enough for some projects, especially those in which requirements are not known at the beginning or may change during development. For a given project, a spiral process may cost more and take more time than does a waterfall process, but may better satisfy a customer's expectations and needs. Models for simulating various waterfall processes have been developed previously, but until now, there have been no models for simulating spiral processes. The present spiral-process-simulating model and the software that implements it were developed by extending a discrete-event simulation process model of the IEEE 12207 Software Development Process, which was built using commercially available software known as the Process Analysis Tradeoff Tool (PATT). Typical inputs to PATT models include industry-average values of product size (expressed as number of lines of code), productivity (number of lines of code per hour), and number of defects per source line of code. The user provides the number of resources, the overall percent of effort that should be allocated to each process step, and the number of desired staff members for each step. The output of PATT includes the size of the product, a measure of effort, a measure of rework effort, the duration of the entire process, and the numbers of injected, detected, and corrected defects as well as a number of other interesting features. In the development of the present model, steps were added to the IEEE 12207 waterfall process, and this model and its implementing software were made to run repeatedly through the sequence of steps, each repetition representing an iteration in a spiral process. Because the IEEE 12207 model is founded on a waterfall paradigm, it enables direct comparison of spiral and waterfall processes. The model can be used throughout a software-development project to analyze the project as more information becomes available. For instance, data from early iterations can be used as inputs to the model, and the model can be used to estimate the time and cost of carrying the project to completion.

Mizell, Carolyn↗

Variations in Sleep and Performance by Duty Start Time in Short Haul Operations

Prior studies have confirmed that commercial airline pilots experience circadian phase shifts and short sleep duration following travel with layovers in different time zones. Few studies have examined the impact of early and late starts on the sleep and circadian phase of airline pilots who return to their domicile after each duty period. We recruited 44 pilots (4 female) from a short-haul commercial airline to participate in a study examining sleep and circadian phase over four duty schedules (baseline, early starts, mid-day starts, late starts). Each duty schedule was five days long, separated by three rest days. Participants completed the rosters in the same order. Sleep outcomes were estimated using wrist-borne actigraphy (Actiware Software, Respironics, Bend, OR) and daily sleep diaries. Thirteen participants volunteered to collect urine samples for the assessment of 6-sulfatoxymelatonin (aMT6s). Urine samples were collected in four-hourly bins during the day and eight-hourly bins during sleep episodes, for 24 hours immediately following each experimental duty schedule. The aMT6s results were fit to a cosine in order to obtain the acrophase to estimate circadian phase. Univariate statistics were calculated for acrophase changes, schedule start times and sleep times. All statistical analyses were computed using SAS software (Cary, IN).

fatigue↗

Variation in Sleep Duration and Circadian Phase by Duty Start Time Among Short-Haul Commercial Airline Pilots

Prior studies have confirmed that commercial airline pilots experience circadian phase shifts and short sleep duration following travel with layovers in different time zones. Few studies have examined the impact of early and late starts on the sleep and circadian phase of airline pilots who return to their domicile after each duty period. We recruited 44 pilots (4 female) from a short-haul commercial airline to participate in a study examining sleep and circadian phase over four duty schedules (baseline, early starts, mid-day starts, late starts). Each duty schedule was five days long, separated by three rest days. Participants completed the rosters in the same order. Sleep outcomes were estimated using wrist-borne actigraphy (Actiware Software, Respironics, Bend, OR) and daily sleep diaries. Thirteen participants volunteered to collect urine samples for the assessment of 6-sulfatoxymelatonin (aMT6s). Urine samples were collected in four hourly bins during the day and eight-hourly bins during sleep episodes, for 24 hours immediately following each experimental duty schedule. The aMT6s results were fit to a cosine in order to obtain the acrophase to estimate circadian phase. Univariate statistics were calculated for acrophase changes, schedule start times and sleep times. All statistical analyses were computed using SAS software (Cary, IN).The mean duty start times varied as expected (baseline 10:17 [ 3:50], early starts 5:24 [ 0:30], mid-day starts 13:52 [ 1:20], late starts 16:33 [ 1:33]). Actigraphy-derived sleep duration varied between schedule types, with the shortest average sleep durations occurring during the early starts and night duty. Mean circadian phase was similar during each schedule type (baseline 26:14 [ 3:22], early starts 25:29 [2:13], mid-day starts 26:20 [ 3:16], late starts 25:49 [ 4:28]), but there were wide inter-individual differences in phase shifting from the beginning to the end of the study, with phase shifts ranging from a 5.98 hour phase advance to an 11.34 hour phase delay. Our preliminary findings suggest that early and late starts are associated with reduced sleep duration. The dispersion in inter-individual differences in circadian phase across shifting duty schedules should be further evaluated to determine how schedule-induced phase shifts may affect operational performance.

Flynn-Evans, Erin↗

Validation of temperature retrievals obtained by the Limb Infrared Monitor of the Stratosphere (LIMS) experiment on Nimbus 7

An outline is given of LIMS temperature determinations (as a function of pressure) from measurements in two channels covering portions of the 15-micrometer band of carbon dioxide. The known sources of error from the radiometer and data reduction are used to estimate the systematic and random errors expected of the results. Observational determinations of the complete end-to-end precision are obtained by computing the standard deviation of six sequential temperature retrievals in regions where the atmosphere is horizontally uniform. This yields values of 0.2 to 0.6 K, in reasonable agreement with the estimates. A correction for horizontal gradients in the atmosphere leads to a large reduction in the differences between the stratospheric temperatures determined on the ascending and descending portions of the orbit. The temperatures agree in the mean with radiosondes and rocketsondes to within 1-2 K in most regions below 1 mbar. Several interesting, previously unseen features included cold regions in the mid-latitude mesosphere and wavelike vertical variations in the tropics.

Gille, J. C.↗

Habitability Assessments And Lessons-learned From 3-day And 11-day Enriched Oxygen Hypobaric Chamber Tests At NASA Johnson Space Center

INTRODUCTION: Decompression sickness (DCS) is a risk to the health and performance of astronauts and high-altitude aircrew. Tolerance to flammability, hypoxia, prebreathe duration, and DCS risk varies across different organizations, vehicles, suits, and destinations, necessitating a variety of DCS risk mitigation approaches. Existing models of altitude DCS risk are often insufficient to enable accurate risk-informed decisions during hardware development, mission planning, and flight operations. METHODS: NASA completed outfitting of a dedicated facility at Johnson Space Center to support testing of up to eight human subjects for multiple days in hypobaric and enriched oxygen atmospheres. The primary purpose of the testing capability is validation of DCS risk mitigation protocols for Artemis missions to the Moon; however, it will also support development and validation of a generalizable altitude DCS risk estimation tool. A 3-day and an 11-day prebreathe validation test were completed in 2022, each with 8 human subjects living at 56.5 kPa (8.2 psia), 34% O2, 66% N2, with 5 simulated EVAs performed on masks at 29.6 kPa (4.3 psi), 85% O2, 15% N2. Facility and organizational lessons-learned and process improvements were recorded during and following the tests, and subjective habitability ratings were recorded daily during the 11-day test. Hypoxia and DCS-related physiological and cognitive outcome measures were recorded during both tests and are reported in companion presentations. RESULTS & DISCUSSION: All subjects completed each of the tests. Primary habitability issues related to mask discomfort during simulated EVAs and poor sleep quality due to thin mattresses. Polybenzimidazole (PBI) clothing was worn by all subjects due to the increased fire risk and may be required for Artemis missions; clothing was found to be acceptable overall with the worst ratings being due to poor fit and inelasticity. Chamber O2 and CO2 sensor inconsistency was observed that did not result in test termination but required post-test follow-up. Forward plans include additional hypobaric testing and integration of existing and future physiological outcome data into an open-source Aerospace Estimation Tool for Hypobaric Exposure Risk (AETHER). NASA is also working to make the testing capability available to commercial companies.

Andrew F J Abercromby↗

Habitability Assessments And Lessons-learned From 3-day And 11-day Enriched Oxygen Hypobaric Chamber Tests At NASA Johnson Space Center

INTRODUCTION: Decompression sickness (DCS) is a risk to the health and performance of astronauts and high-altitude aircrew. Tolerance to flammability, hypoxia, prebreathe duration, and DCS risk varies across different organizations, vehicles, suits, and destinations, necessitating a variety of DCS risk mitigation approaches. Existing models of altitude DCS risk are often insufficient to enable accurate risk-informed decisions during hardware development, mission planning, and flight operations. METHODS: NASA completed outfitting of a dedicated facility at Johnson Space Center to support testing of up to eight human subjects for multiple days in hypobaric and enriched oxygen atmospheres. The primary purpose of the testing capability is validation of DCS risk mitigation protocols for Artemis missions to the Moon; however, it will also support development and validation of a generalizable altitude DCS risk estimation tool. A 3-day and an 11-day prebreathe validation test were completed in 2022, each with 8 human subjects living at 56.5 kPa (8.2 psia), 34% O2, 66% N2, with 5 simulated EVAs performed on masks at 29.6 kPa (4.3 psi), 85% O2, 15% N2. Facility and organizational lessons-learned and process improvements were recorded during and following the tests, and subjective habitability ratings were recorded daily during the 11-day test. Hypoxia and DCS-related physiological and cognitive outcome measures were recorded during both tests and are reported in companion presentations. RESULTS & DISCUSSION: All subjects completed each of the tests. Primary habitability issues related to mask discomfort during simulated EVAs and poor sleep quality due to thin mattresses. Polybenzimidazole (PBI) clothing was worn by all subjects due to the increased fire risk and may be required for Artemis missions; clothing was found to be acceptable overall with the worst ratings being due to poor fit and inelasticity. Chamber O2 and CO2 sensor inconsistency was observed that did not result in test termination but required post-test follow-up. Forward plans include additional hypobaric testing and integration of existing and future physiological outcome data into an open-source Aerospace Estimation Tool for Hypobaric Exposure Risk (AETHER). NASA is also working to make the testing capability available to commercial companies.

Andrew Abercromby↗

Methods for Characterizing Fine Particulate Matter Using Satellite Remote-Sensing Data and Ground Observations: Potential Use for Environmental Public Health Surveillance

This study describes and demonstrates different techniques for surfacing daily environmental / hazards data of particulate matter with aerodynamic diameter less than or equal to 2.5 micrometers (PM2.5) for the purpose of integrating respiratory health and environmental data for the Centers for Disease Control and Prevention (CDC s) pilot study of Health and Environment Linked for Information Exchange (HELIX)-Atlanta. It described a methodology for estimating ground-level continuous PM2.5 concentrations using B-Spline and inverse distance weighting (IDW) surfacing techniques and leveraging National Aeronautics and Space Administration (NASA) Moderate Resolution Imaging Spectrometer (MODIS) data to complement The Environmental Protection Agency (EPA) ground observation data. The study used measurements of ambient PM2.5 from the EPA database for the year 2003 as well as PM2.5 estimates derived from NASA s satellite data. Hazard data have been processed to derive the surrogate exposure PM2.5 estimates. The paper has shown that merging MODIS remote sensing data with surface observations of PM2.5 not only provides a more complete daily representation of PM2.5 than either data set alone would allow, but it also reduces the errors in the PM2.5 estimated surfaces. The results of this paper have shown that the daily IDW PM2.5 surfaces had smaller errors, with respect to observations, than those of the B-Spline surfaces in the year studied. However the IDW mean annual composite surface had more numerical artifacts, which could be due to the interpolating nature of the IDW that assumes that the maxima and minima can occur only at the observation points. Finally, the methods discussed in this paper improve temporal and spatial resolutions and establish a foundation for environmental public health linkage and association studies for which determining the concentrations of an environmental hazard such as PM2.5 with good accuracy levels is critical.

Al-Hamdan, Mohammad Z.↗

BUCKLING OF A SPHERE OF EXTREMELY HIGH RADIUS-THICKNESS RATIO

Proposed passive communications satellites are ultrathin-walled spherical shells which may experience buckling from external pressure due to solar radiation or atmospheric drag. Large buckling deformations of such a satellite are undesirable because they seriously reduce the satellite’s performance as a reflector. At present, shell buckling technology is lacking in several respects with regard to design of spherical shells subjected to external pressure. Large discrepancies exist between theoretical and experimental buckling pressure; therefore, the designer must be guided primarily by experimental data. However, very little experimental data on complete spheres are available; the primary research effort has been concerned with shallow spherical caps. All available experimental data are for radius-thickness ratios several orders of magnitude smaller than that for the proposed satellites. In order to provide a basis for estimating the buckling strength of these satellites, therefore, buckling experiments were conducted on a complete spherical shell having a radius-thickness ratio of about 85,000. This value is much higher than that of any spherical cap or complete sphere used in previously reported experiments.

SHELL STABILITY↗

Comparison of Two Methods for Noninvasive Determination of Stroke Volume During Orthostatic Challenge

Background: The real time, beat-by-beat, non-invasive determination of stroke volume (SV) is an important parameter in many aerospace related physiologic protocols. In this study, we compared simultaneous estimates of SV calculated from peripheral pulse waveforms with a more conventional non-invasive technique. Methods: Using a prospective, randomized blinded protocol, ten males and nine females completed 12-mm tilt table protocols. The relative change (%(Delta)) in beat-to-beat SV was estimated non-invasively from changes in pulse waveforms measured by application of infrared finger photoplethysmography (IFP) with a Portapres(Registered TradeMark) blood pressure monitoring device and by thoracic impedance cardiography (TIC). The %(Delta) SV values were calculated from continuous SV measurements in the supine posture and over the first 10 s (T1), second 10 s (T2), and 3.5 minutes (T3) of 80deg head-up tilt (HUT). Results: The average %(Delta) SV measured by IFP at T1 (-11.7 +/- 3.7 %) was statistically less (P <0.02) than the average %(Delta) SV measured by TIC at T1 (-21.7 +/- 3.1 %), while the average %(Delta) SV measured by 1FF at T2 (-16.2 +/- 3.9 %) and T3 (-19.1 +/- 3.8 %) were not statistically distinguishable (P > or = 0.322) than the average %(Delta) SV measured by TIC at T2 (-21.8 +/- 2.5 %), and T3 (-22.6 +/- 2.9 %). Correlation coefficients (r(sup 2)) between IFP and TIC were 0.117 (T1), 0.387 (T2), and 0.7 18 (T3). Conclusion: IFP provides beat-to-beat (real time) assessment of %(Delta) SV after 20 sec of transition to an orthostatic challenge that is comparable to the commonly accepted TIC. Our data support the notion that IFP technology which has flown during space missions can be used to accurately assess physiological status and countermeasure effectiveness for orth static problems that may arise in astronauts after space flight. While the peripherally measured IFP response is slightly delayed, the ease of implementing this monitor in the field is advantageous.

Doerr, Donald F.↗

Estimation of duration and mental workload at differing times of day by males and females

Two experiments are reported which investigated whether male and female operator duration estimation and subjective workload followed conventional circadian fluctuation. In the first experiment, twenty-four subjects performed a filled time-estimation task in a constant blacked-out, noise-reduced environment at 0800, 1200, 1600, and 2000 h. In the second experiment, twelve subjects performed an unfilled time estimation task in similar conditions at 0900, 1400, and 1900 h. At the termination of all experimental sessions, participants completed the NASA TLX workload assessment questionnaire as a measure of perceived mental workload. Results indicated that while physiological response followed an expected pattern, estimations of duration and subjective perception of workload showed no significant effects for time-of-day. In each of the experiments, however, there were significant differences in durational estimates and mental workload response depending upon the gender of the participant. Results are taken to support the assertion that subjective workload is responsive largely to task-related factors and indicates the important differences that may be expected due to operator gender.

Hancock, P. A.↗

Adaptation of a Fast Optimal Interpolation Algorithm to the Mapping of Oceangraphic Data

A fast, recently developed, multiscale optimal interpolation algorithm has been adapted to the mapping of hydrographic and other oceanographic data. This algorithm produces solution and error estimates which are consistent with those obtained from exact least squares methods, but at a small fraction of the computational cost. Problems whose solution would be completely impractical using exact least squares, that is, problems with tens or hundreds of thousands of measurements and estimation grid points, can easily be solved on a small workstation using the multiscale algorithm. In contrast to methods previously proposed for solving large least squares problems, our approach provides estimation error statistics while permitting long-range correlations, using all measurements, and permitting arbitrary measurement locations. The multiscale algorithm itself, published elsewhere, is not the focus of this paper. However, the algorithm requires statistical models having a very particular multiscale structure; it is the development of a class of multiscale statistical models, appropriate for oceanographic mapping problems, with which we concern ourselves in this paper. The approach is illustrated by mapping temperature in the northeastern Pacific. The number of hydrographic stations is kept deliberately small to show that multiscale and exact least squares results are comparable. A portion of the data were not used in the analysis; these data serve to test the multiscale estimates. A major advantage of the present approach is the ability to repeat the estimation procedure a large number of times for sensitivity studies, parameter estimation, and model testing. We have made available by anonymous Ftp a set of MATLAB-callable routines which implement the multiscale algorithm and the statistical models developed in this paper.

Menemenlis, Dimitris↗

Predictability of the California Current System

The physical and biological oceanography of the Southern California Bight (SCB), a highly productive subregion of the California Current System (CCS) that extends from Point Conception, California, south to Ensenada, Mexico, continues to be extensively studied. For example, the California Cooperative Oceanic Fisheries Investigations (CalCOFI) program has sampled this region for over 50 years, providing an unparalleled time series of physical and biological data. However, our understanding of what physical processes control the large-scale and mesoscale variations in these properties is incomplete. In particular, the non-synoptic and relatively coarse spatial sampling (70km) of the hydrographic grid does not completely resolve the mesoscale eddy field (Figure 1a). Moreover, these unresolved physical variations exert a dominant influence on the evolution of the ecosystem. In recent years, additional datasets that partially sample the SCB have become available. Acoustic Doppler Current Profiler (ADCP) measurements, which now sample upper-ocean velocity between stations, and sea level observations along TOPEX tracks give a more complete picture of the mesoscale variability. However, both TOPEX and ADCP are well-sampled only along the cruise or orbit tracks and coarsely sampled in time and between tracks. Surface Lagrangian drifters also sample the region, although irregularly in time and space. SeaWiFS provides estimates of upper-ocean chlorophyll-a (chl-alpha), usually giving nearly complete coverage for week-long intervals, depending on cloud coverage. Historical ocean color data from the Coastal Zone Color Scanner (CZCS) has been used extensively to determine phytoplankton patterns and variability, characterize the primary production across the SCB coastal fronts, and describe the seasonal and interannual variability in pigment concentrations. As in CalCOFI, these studies described much of the observed structures and their variability over relatively large space and time scales.

Miller, Arthur J.↗

Detailed Design and Cost Estimation of a 300 MWe Oxy-Fuel sCO2 Turbine

The detailed design of a 300 MWe, utility scale oxy-fuel turbine has been completed for purposed operation in the sCO2 direct fired Allam-Fetvedt cycle, targeting near-zero emissions and a 50% LHV system efficiency. The turbine and its supporting plant aim to offer a lower levelized cost of energy than a natural gas combined cycle plant employing carbon capture. The oxy-fuel turbine conditions include an inlet temperature of 1150°C and inlet pressure of 305 bar, representing temperatures near that of a gas turbine simultaneously with pressures near an ultra-supercritical steam turbine. The combustor housing and turbine designs were completed according to the ASME BPVC; the turbine case specifically incorporates a multi-body design with inner high-pressure barrel case and low-pressure (30 bar) horizontally split outer case of low-chromium steel material. Lateral rotordynamic evaluation demonstrated acceptable vibration response for a range of imbalance conditions per API standards. The cooling flow required in the six-stage turbine flowpath for 30,000 hr. blade and stator lifetime is predicted through thermal and structural modeling of the first stage. The provided cost estimate of the turbine is formed through a combination of scaled up-costs from procured 10 MWe scale sCO2 turbomachinery hardware, and vendor provided budgetary quotes of larger components including the turbine case requiring casting, welding, and final machining processes. The performance and cost estimation of the oxy-fuel turbine predicted for the completed detailed design provides important information towards future development needs for market penetration of utility scale direct fired sCO2 power cycles.

Marshall, Michael [Southwest Research Institute, S↗

Distortion Representation of Forecast Errors for Model Skill Assessment and Objective Analysis

We completed the formulation of the smoothness penalty functional this past quarter. We used a simplified procedure for estimating the statistics of the FCA solution spectral coefficients from the results of the unconstrained, low-truncation FCA (stopping criterion) solutions. During the current reporting period we have completed the calculation of GEOS-2 model-equivalent brightness temperatures for the 6.7 micron and 11 micron window channels used in the GOES imagery for all 10 cases from August 1999. These were simulated using the AER-developed Optimal Spectral Sampling (OSS) model.

Hoffman, Ross N.↗

Gas-Phase Reaction Pathways and Rate Coefficients for the Dichlorosilane-Hydrogen and Trichlorosilane-Hydrogen Systems

As part of NASA Ames Research Center's Integrated Process Team on Device/Process Modeling and Nanotechnology our goal is to create/contribute to a gas-phase chemical database for use in modeling microelectronics devices. In particular, we use ab initio methods to determine chemical reaction pathways and to evaluate reaction rate coefficients. Our initial studies concern reactions involved in the dichlorosilane-hydrogen (SiCl2H2--H2) and trichlorosilane-hydrogen (SiCl2H-H2) systems. Reactant, saddle point (transition state), and product geometries and their vibrational harmonic frequencies are determined using the complete-active-space self-consistent-field (CASSCF) electronic structure method with the correlation consistent polarized valence double-zeta basis set (cc-pVDZ). Reaction pathways are constructed by following the imaginary frequency mode of the saddle point to both the reactant and product. Accurate energetics are determined using the singles and doubles coupled-cluster method that includes a perturbational estimate of the effects of connected triple excitations (CCSD(T)) extrapolated to the complete basis set limit. Using the data from the electronic structure calculations, reaction rate coefficients are obtained using conventional and variational transition state and RRKM theories.

Dateo, Christopher E.↗

The power spectrum of galaxies in the nearby universe

We compute the power spectrum of galaxy density fluctuations in a recently completed Southern Sky Redshift Survey of optically selected galaxies (SSRS2). The amplitude and shape of the SSRS2 power spectrum are consistent with results of the Center for Astrophysics redshift survey of the northern hemisphere (CfA2), including the abrupt change of slope on a scale of 30-50/h Mpc; these results are reproducible for independent volumes of space, and variations are consistent with the errors estimated from mock surveys. Taken together, the SSRS2 and the CfA2 form a complete sample of 14,383 galaxies which covers one-third of the sky. The power spectrum of this larger sample continues to rise on scales up to approximately 200/h Mpc, with weak evidence for flattening on the largest scales. The SSRS2 + CfA2 power spectrum and the power spectrum constraints implied by COBE are well matched by an Omega(h) is approximately 0.2, Omega + lambda(sub 0) = 1 cold dark matter model with minimal biasing of optically selected galaxies.

Da Costa, L. Nicolaci↗

Decomposing a San Francisco estuary microbiome using long-read metagenomics reveals species- and strain-level dominance from picoeukaryotes to viruses

ABSTRACT Although long-read sequencing has enabled obtaining high-quality and complete genomes from metagenomes, many challenges still remain to completely decompose a metagenome into its constituent prokaryotic and viral genomes. This study focuses on decomposing an estuarine metagenome to obtain a more accurate estimate of microbial diversity. To achieve this, we developed a new bead-based DNA extraction method, a novel bin refinement method, and obtained 150 Gbp of Nanopore sequencing. We estimate that there are ~500 bacterial and archaeal species in our sample and obtained 68 high-quality bins (>90% complete, <5% contamination, ≤5 contigs, contig length of >100 kbp, and all ribosomal and tRNA genes). We also obtained many contigs of picoeukaryotes, environmental DNA of larger eukaryotes such as mammals, and complete mitochondrial and chloroplast genomes and detected ~40,000 viral populations. Our analysis indicates that there are only a few strains that comprise most of the species abundances. IMPORTANCE Ocean and estuarine microbiomes play critical roles in global element cycling and ecosystem function. Despite the importance of these microbial communities, many species still have not been cultured in the lab. Environmental sequencing is the primary way the function and population dynamics of these communities can be studied. Long-read sequencing provides an avenue to overcome limitations of short-read technologies to obtain complete microbial genomes but comes with its own technical challenges, such as needed sequencing depth and obtaining high-quality DNA. We present here new sampling and bioinformatics methods to attempt decomposing an estuarine microbiome into its constituent genomes. Our results suggest there are only a few strains that comprise most of the species abundances from viruses to picoeukaryotes, and to fully decompose a metagenome of this diversity requires 1 Tbp of long-read sequencing. We anticipate that as long-read sequencing technologies continue to improve, less sequencing will be needed.

Lui, Lauren M.↗