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A near-complete Pseudomonas stutzeri draft genome was extracted from a coalbed metagenome. The draft genome described herein provides insight into the functional pathways encoded by this bacterium and its potential role in coalbed methane environments.
The “Candidatus Synechococcus spongiarum” group includes different clades of cyanobacteria with high 16S rRNA sequence identity (~99%) and is the most abundant and widespread cyanobacterial symbiont of marine sponges. The first draft genome of a “Ca. Synechococcus spongiarum” group member was recently published, providing evidence of genome reduction by loss of genes involved in several nonessential functions. However, “Ca. Synechococcus spongiarum” includes a variety of clades that may differ widely in genomic repertoire and consequently in physiology and symbiotic function. Here, we present three additional draft genomes of “Ca. Synechococcus spongiarum,” each from a different clade. By comparing all four symbiont genomes to those of free-living cyanobacteria, we revealed general adaptations to life inside sponges and specific adaptations of each phylotype. Symbiont genomes shared about half of their total number of coding genes. Common traits of “Ca. Synechococcus spongiarum” members were a high abundance of DNA modification and recombination genes and a reduction in genes involved in inorganic ion transport and metabolism, cell wall biogenesis, and signal transduction mechanisms. Moreover, these symbionts were characterized by a reduced number of antioxidant enzymes and low-weight peptides of photosystem II compared to their free-living relatives. Variability within the “Ca. Synechococcus spongiarum” group was mostly related to immune system features, potential for siderophore-mediated iron transport, and dependency on methionine from external sources. The common absence of genes involved in synthesis of residues, typical of the O antigen of free-living Synechococcus species, suggests a novel mechanism utilized by these symbionts to avoid sponge predation and phage attack.
This software implements the messages defined in the ISO 15118-202 standard within the Everest EV Charging open source framework. The protocol and messages defined in the ISO 15118-202 standard enable the exchange of additional information which is not available for exchange within the currently deployed EV/EVSE communications protocols. This information includes co-identification parameters, error message exchange and more. This fork of the everest-core repository adds a prototype of the Extensible Supply Equipment Communication Controller (SECC) Discovery Protocol (ESDP) implemented based on a draft of the ISO 15118-202 standard. This is achieved through additions and modifications to the EvseV2G module. The implementation provides a demonstration of the ESDP messages, encoding and decoding but does not include a full integration within the Everest framework. Much of the information being sent over ESDP in this implementation is set statically for the sake of demonstrating the protocol itself. This fork of the ext-switchev-iso15118 repository adds a prototype of the Extensible Supply Equipment Communication Controller (SECC) Discovery Protocol (ESDP) implemented based on a draft of the ISO 15118-202 standard. The implementation provides a demonstration of the ESDP messages, encoding and decoding but does not include a full integration within the Everest framework. Much of the information being sent over ESDP in this implementation is set statically for the sake of demonstrating the protocol itself. This fork adds the ESDP features for only the EVCC controller because that is the only portion that is utilized in the everest Software-in-the-Loop.
Background Clostridium autoethanogenum strain JA1-1 (DSM 10061) is an acetogen capable of fermenting CO, CO2 and H2 (e.g. from syngas or waste gases) into biofuel ethanol and commodity chemicals such as 2,3-butanediol. A draft genome sequence consisting of 100 contigs has been published. Results A closed, high-quality genome sequence for C. autoethanogenum DSM10061 was generated using only the latest single-molecule DNA sequencing technology and without the need for manual finishing. It is assigned to the most complex genome classification based upon genome features such as repeats, prophage, nine copies of the rRNA gene operons. It has a low G + C content of 31.1%. Illumina, 454, Illumina/454 hybrid assemblies were generated and then compared to the draft and PacBio assemblies using summary statistics, CGAL, QUAST and REAPR bioinformatics tools and comparative genomic approaches. Assemblies based upon shorter read DNA technologies were confounded by the large number repeats and their size, which in the case of the rRNA gene operons were ~5 kb. CRISPR (Clustered Regularly Interspaced Short Paloindromic Repeats) systems among biotechnologically relevant Clostridia were classified and related to plasmid content and prophages. Potential associations between plasmid content and CRISPR systems may have implications for historical industrial scale Acetone-Butanol-Ethanol (ABE) fermentation failures and future large scale bacterial fermentations. While C. autoethanogenum contains an active CRISPR system, no such system is present in the closely related Clostridium ljungdahlii DSM 13528. A common prophage inserted into the Arg-tRNA shared between the strains suggests a common ancestor. However, C. ljungdahlii contains several additional putative prophages and it has more than double the amount of prophage DNA compared to C. autoethanogenum. Other differences include important metabolic genes for central metabolism (as an additional hydrogenase and the absence of a phophoenolpyruvate synthase) and substrate utilization pathway (mannose and aromatics utilization) that might explain phenotypic differences between C. autoethanogenum and C. ljungdahlii. Conclusions Single molecule sequencing will be increasingly used to produce finished microbial genomes. The complete genome will facilitate comparative genomics and functional genomics and support future comparisons between Clostridia and studies that examine the evolution of plasmids, bacteriophage and CRISPR systems.
Strains of a pink-pigmented Methylobacterium sp. are effective nitrogen- (N2) fixing microsymbionts of species of the African crotalarioid genus Listia. Strain WSM2598 is an aerobic, motile, Gram-negative, non-spore-forming rod isolated in 2002 from a Listia bainesii root nodule collected at Estcourt Research Station in South Africa. Here we describe the features of Methylobacterium sp. WSM2598, together with information and annotation of a high-quality draft genome sequence. The 7,669,765 bp draft genome is arranged in 5 scaffolds of 83 contigs, contains 7,236 protein-coding genes and 18 RNA-only encoding genes. This rhizobial genome is one of 100 sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project.
Recent advances in high-volume sequencing technology and mining of genomes from metagenomic samples call for rapid and reliable genome quality evaluation. The current release of the PATRIC database contains over 220,000 genomes, and current metagenomic technology supports assemblies of many draft-quality genomes from a single sample, most of which will be novel. We have added two quality assessment tools to the PATRIC annotation pipeline. EvalCon uses supervised machine learning to calculate an annotation consistency score. EvalG implements a variant of the CheckM algorithm to estimate contamination and completeness of an annotated genome.We report on the performance of these tools and the potential utility of the consistency score. Additionally, we provide contamination, completeness, and consistency measures for all genomes in PATRIC and in a recent set of metagenomic assemblies. EvalG and EvalCon facilitate the rapid quality control and exploration of PATRIC-annotated draft genomes.
Background: Clostridium botulinum is a diverse group of bacteria characterized by the production of botulinum neurotoxin. Botulinum neurotoxins are classified into serotypes (BoNT/A–G), which are produced by six species/Groups of Clostridia , but the genetic background of the bacteria remains poorly understood. The purpose of this study was to use comparative genomics to provide insights into the genetic diversity and evolutionary history of bacteria that produce the potent botulinum neurotoxin. Results: Comparative genomic analyses of over 170 Clostridia genomes, including our draft genome assemblies for 59 newly sequenced Clostridia strains from six continents and publicly available genomic data, provided in-depth insights into the diversity and distribution of BoNT-producing bacteria. These newly sequenced strains included Group I and II strains that express BoNT/A,/B,/E, or/F as well as bivalent strains. BoNT-producing Clostridia and closely related Clostridia species were delineated with a variety of methods including 16S rRNA gene, concatenated marker genes, core genome and concatenated multi-locus sequencing typing (MLST) gene phylogenies that related whole genome sequenced strains to publicly available strains and sequence types. These analyses illustrated the phylogenetic diversity in each Group and the diversity of genomic backgrounds that express the same toxin type or subtype. Comparisons of the botulinum neurotoxin genes did not identify novel toxin types or variants. Conclusions: This study represents one of the most comprehensive analyses of whole genome sequence data for Group I and II BoNT-producing strains. Read data and draft genome assemblies generated for 59 isolates will be a resource to the research community. Core genome phylogenies proved to be a powerful tool for differentiating BoNT-producing strains and can provide a framework for the study of these bacteria. Comparative genomic analyses of Clostridia species illustrate the diversity of botulinum-neurotoxin-producing strains and the plasticity of the genomic backgrounds in which bont genes are found.
Background Glaciers cover ~ 10% of land but are among the least explored environments on Earth. The basal portion of glaciers often harbors unique aquatic microbial ecosystems in the absence of sunlight, and knowledge on the microbial community structures and their metabolic potential is very limited. Here, we provide insights into the microbial lifestyle present at the base of the Matanuska Glacier, Alaska. Results DNA and RNA were extracted from samples of the Matanuska Glacier basal ice. Using Illumina MiSeq and HiSeq sequencing, we investigated the microbial diversity with the metagenomic shotgun reads and 16S ribosomal RNA data. We further assembled 9 partial and draft bacterial genomes from the metagenomic assembly, and identified key metabolic pathways such as sulfur oxidation and nitrification. Collectively, our analyses suggest a prevalence of lithotrophic and heterotrophic metabolisms in the subglacial microbiome. Conclusion Our results present the first metagenomic assembly and bacterial draft genomes for a subglacial environment. These results extend our understanding of the chemical and biological processes in subglacial environments critically influenced by global climate change.
Polycyclic aromatic hydrocarbons (PAH) are ubiquitous environmental pollutants and microbial biodegradation is an important means of remediation of PAH-contaminated soil. Delftia acidovorans Cs1-4 (formerly Delftia sp. Cs1-4) was isolated by using phenanthrene as the sole carbon source from PAH contaminated soil in Wisconsin. Its full genome sequence was determined to gain insights into a mechanisms underlying biodegradation of PAH. Three genomic libraries were constructed and sequenced: an Illumina GAii shotgun library (916,416,493 reads), a 454 Titanium standard library (770,171 reads) and one paired-end 454 library (average insert size of 8 kb, 508,092 reads). The initial assembly contained 40 contigs in two scaffolds. The 454 Titanium standard data and the 454 paired end data were assembled together and the consensus sequences were computationally shredded into 2 kb overlapping shreds. Illumina sequencing data was assembled, and the consensus sequence was computationally shredded into 1.5 kb overlapping shreds. Gaps between contigs were closed by editing in Consed, by PCR and by Bubble PCR primer walks. A total of 182 additional reactions were needed to close gaps and to raise the quality of the finished sequence. The final assembly is based on 253.3 Mb of 454 draft data (averaging 38.4 X coverage) and 590.2 Mb of Illumina draft data (averaging 89.4 X coverage). The genome of strain Cs1-4 consists of a single circular chromosome of 6,685,842 bp (66.7 %G+C) containing 6,028 predicted genes; 5,931 of these genes were protein-encoding and 4,425 gene products were assigned to a putative function. Genes encoding phenanthrene degradation were localized to a 232 kb genomic island (termed the phn island), which contained near its 3’ end a bacteriophage P4-like integrase, an enzyme often associated with chromosomal integration of mobile genetic elements. Other biodegradation pathways reconstructed from the genome sequence included: benzoate (by the acetyl-CoA pathway), styrene, nicotinic acid (by the maleamate pathway) and the pesticides Dicamba and Fenitrothion. Lastly, determination of the complete genome sequence of D. acidovorans Cs1-4 has provided new insights the microbial mechanisms of PAH biodegradation that may shape the process in the environment.
Sulfurospirillum spp. play an important role in sulfur and nitrogen cycling, and contain metabolic versatility that enables reduction of a wide range of electron acceptors, including thiosulfate, tetrathionate, polysulfide, nitrate, and nitrite. Here we describe the assembly of a Sulfurospirillum genome obtained from the metagenome of an electrosynthetic microbiome. The ubiquity and persistence of this organism in microbial electrosynthesis systems suggest it plays an important role in reactor stability and performance. Understanding why this organism is present and elucidating its genetic repertoire provide a genomic and ecological foundation for future studies where Sulfurospirillum are found, especially in electrode-associated communities. Metabolic comparisons and in-depth analysis of unique genes revealed potential ecological niche-specific capabilities within the Sulfurospirillum genus. The functional similarities common to all genomes, i.e., core genome, and unique gene clusters found only in a single genome were identified. Based upon 16S rRNA gene phylogenetic analysis and average nucleotide identity, the Sulfurospirillum draft genome was found to be most closely related to Sulfurospirillum cavolei. Characterization of the draft genome described herein provides pathway-specific details of the metabolic significance of the newly described Sulfurospirillum cavolei MES and, importantly, yields insight to the ecology of the genus as a whole. Comparison of eleven sequenced Sulfurospirillum genomes revealed a total of 6246 gene clusters in the pan-genome. Of the total gene clusters, 18.5% were shared among all eleven genomes and 50% were unique to a single genome. While most Sulfurospirillum spp. reduce nitrate to ammonium, five of the eleven Sulfurospirillum strains encode for a nitrous oxide reductase (nos) cluster with an atypical nitrous-oxide reductase, suggesting a utility for this genus in reduction of the nitrous oxide, and as a potential sink for this potent greenhouse gas.
The colonial green microalga Botryococcus braunii is well known for producing liquid hydrocarbons that can be utilized as biofuel feedstocks. B. braunii is taxonomically classified as a single species made up of three chemical races, A, B, and L, that are mainly distinguished by the hydrocarbons produced. We previously reported a B race draft nuclear genome, and here we report the draft nuclear genomes for the A and L races. A comparative genomic study of the three B. braunii races and 14 other algal species within Chlorophyta revealed significant differences in the genomes of each race of B. braunii. Phylogenomically, there was a clear divergence of the three races with the A race diverging earlier than both the B and L races, and the B and L races diverging from a later common ancestor not shared by the A race. DNA repeat content analysis suggested the B race had more repeat content than the A or L races. Orthogroup analysis revealed the B. braunii races displayed more gene orthogroup diversity than three closely related Chlamydomonas species, with nearly 24-36% of all genes in each B. braunii race being specific to each race. This analysis suggests the three races are distinct species based on sufficient differences in their respective genomes. We propose reclassification of the three chemical races to the following species names: Botryococcus alkenealis (A race), Botryococcus braunii (B race), and Botryococcus lycopadienor (L race).
The Islands of Martha's Vineyard and Nantucket are separated from the Massachusetts mainland by Vineyard and Nantucket Sounds; water between the two islands flows through Muskeget Channel. The towns of Edgartown (on Martha's Vineyard) and Nantucket recognize that they are vulnerable to power supply interruptions due to their position at the end of the power grid, and due to sea level rise and other consequences of climate change. The tidal energy flowing through Muskeget Channel has been identified by the Electric Power Research Institute as the strongest tidal resource in Massachusetts waters. The Town of Edgartown proposes to develop an initial 5 MW (nameplate) tidal energy project in Muskeget Channel. The project will consist of 14 tidal turbines with 13 providing electricity to Edgartown and one operated by the University of Massachusetts at Dartmouth for research and development. Each turbine will be 90 feet long and 50 feet high. The electricity will be brought to shore by a submarine cable buried 8 feet below the seabed surface which will landfall in Edgartown either on Chappaquiddack or at Katama. Muskeget Channel is located between Martha's Vineyard and Nantucket. Its depth ranges between 40 and 160 feet in the deepest portion. It has strong currents where water is transferred between Nantucket Sound and the Atlantic Ocean continental shelf to the south. This makes it a treacherous passage for navigation. Current users of the channel are commercial and recreational fishing, and cruising boats. The US Coast Guard has indicated that the largest vessel passing through the channel is a commercial scallop dragger with a draft of about 10 feet. The tidal resource in the channel has been measured by the University of Massachusetts-Dartmouth and the peak velocity flow is approximately 5 knots. The technology proposed is the helical Gorlov-type turbine positioned with a horizontal axis that is positively buoyant in the water column and held down by anchors. This is the same technology proposed by Ocean Renewable Power Company in the Western Passage and Cobscook Bay near Eastport Maine. The blades rotate in two directions capturing the tides energy both during flood and ebb tides. The turbines will be anchored to the bottom and suspended in the water column. Initial depth of the turbines is expected to be about 25 feet below the surface to avoid impacting navigation while also capturing the strongest currents. The Town of Edgartown was initially granted a Preliminary Permit by the Federal Energy Regulatory Commission (FERC) on March 1, 2008, and has recently received a second permit valid through August 2014. The Preliminary Permit gives Edgartown the exclusive right to apply for a power generation license for power generated from the hydrokinetic energy in the water flowing in this area. Edgartown filed a Draft Pilot License Application with FERC on February 1, 2010 and an Expanded Environmental Notification Form with the Massachusetts Environmental Policy Act (MEPA) Office at the same time. It expects to file a Final License Application in late 2013. Harris Miller Miller & Hanson (HMMH) of Burlington Massachusetts is acting as the Project Manager for the Town of Edgartown and collaborating with other partners of the project including the University of Massachusetts - Dartmouth's Marine Renewable Energy Center and the Massachusetts Clean Energy Center. HMMH was awarded a grant under the Department of Energy's Advanced Water Program to conduct marine science and hydrokinetic site-specific environmental studies for projects actively seeking a FERC License. HMMH, on behalf of the Town, is managing this comprehensive study of the marine environment in Muskeget Channel and potential impacts of the tidal project on indicator species and habitats. The University of Massachusetts School of Marine Science and Technology (SMAST) conducted oceanographic studies of tidal currents, tide level, benthic habitat, and potential for biofouling and foundation scouring. Woods Hole Oceanographic Institution, cooperating with SMAST, developed an oceanographic model to predict changes in sediment transport as a result of the proposed tidal energy project. Provincetown Center for Coastal Studies prepared background material on protected species - including whales, seals, and sea turtles - in the project area and implemented an initial tagging program to record location specific information on seals and sea turtles. HMMH communicated research plans and findings with local stakeholder groups, state and federal resource agency staff, and the ocean power industry. The information is being used to prepare environmental permit applications and obtain approvals for project construction.
This quarterly report summarizes the status for the project planning to initiate all the legal and contract documents required for establishing the subcontracts needed and a Cooperative Research and Development Agreement (CRADA) with Autodesk, Inc., Toyota Motor Engineering and Manufacturing North America (Toyota), and Magna Exterior and Interiors Corporation (Magna). During the first quarter (10/1/2012 to 12/31/2012), the statements of work (SOW) for the subcontracts to Purdue University, University of Illinois, and PlastiComp, Inc. were completed. A draft of the CRADA SOW was sent to Autodesk, Toyota, and Magna for technical and legal reviews. PNNL Legal Services contacted project partners’ Legal counterparts for preparing legal documents for the project. A non-disclosure agreement was drafted and sent to all the parties for reviews.
Documentation of the energy yield of a large photovoltaic (PV) system over a substantial period can be useful to measure a performance guarantee, as an assessment of the health of the system, for verification of a performance model to then be applied to a new system, or for a variety of other purposes. Although the measurement of this performance metric might appear to be straight forward, there are a number of subtleties associated with variations in weather and imperfect data collection that complicate the determination and data analysis. A performance assessment is most valuable when it is completed with a very low uncertainty and when the subtleties are systematically addressed, yet currently no standard exists to guide this process. This report summarizes a draft methodology for an Energy Performance Evaluation Method, the philosophy behind the draft method, and the lessons that were learned by implementing the method.
The 50th Annual Maize Genetics Conference was held February 27 - March 2, 2008 at the Marriott Wardman Park Hotel in Washington, D.C. As the golden anniversary of the Conference and coinciding with the release of a draft of the maize genome sequence, this was a special meeting. To publicize this unique occasion, meeting organizers hosted a press conference, which was attended by members of the press representing science and non-science publications, and an evening reception at the Smithsonian National Museum of Natural History, where the draft sequence was announced and awards were presented to Dr. Mary Clutter and Senator Kit Bond to thank them for their outstanding contributions to maize genetics and genomics research. As usual, the Conference provided an invigorating forum for exchange of recent research results in many areas of maize genetics, e.g., cytogenetics, development, molecular genetics, transposable element biology, biochemical genetics, and genomics.
A proposed strategy for the remediation of uranium (U) contaminated sites was based on the immobilization of U by reducing the oxidized soluble U, U(VI), to form a reduced insoluble end product, U(IV). Previous studies identified Geobacter sp., including G. sulfurreducens and G. metallireducens, as predominant U(VI)-reducing bacteria under acetate-oxidizing and U(VI)-reducing conditions. Examination of the finished genome sequence annotation of the canonical metal reducing species Geobacter sulfurreducens strain PCA and G. metallireduceans strain GS-15 as well as the draft genome sequence of G. uraniumreducens strain Rf4 identified phage related proteins. In addition, the completed genome for Anaeromyxobacter dehalogenans and the draft genome sequence of Desulfovibrio desulfuricans strain G20, two more model metal-reducing bacteria, also revealed phage related sequences. The presence of these gene sequences indicated that Geobacter spp., Anaeromyxobacter spp., and Desulfovibrio spp. are susceptible to viral infection. Furthermore, viral populations in soils and sedimentary environments in the order of 6.4×10{sup 6}–2.7×10{sup 10} VLP’s cm{sup -3} have been observed. In some cases, viral populations exceed bacterial populations in these environments suggesting that a relationship may exist between viruses and bacteria. Our preliminary screens of samples collected from the ESR FRC indicated that viral like particles were observed in significant numbers. The objective of this study was to investigate the potential functional role viruses play in metal reduction specifically Fe(III) and U(VI) reduction, the environmental parameters affecting viral infection of metal reducing bacteria, and the subsequent effects on U transport.
Isochronous stress-strain curves for Alloy 617 up to a temperature of 1000°C will be required to qualify the material for elevated temperature design in Section III, Division 1, Subsection NH of the ASME Boiler and Pressure Vessel Code. Several potential methods for developing these curves are reviewed in this report. It is shown that in general power-law creep is the rate controlling deformation mechanism for a wide range of alloy heats, test temperatures and stresses. Measurement of the strain rate sensitivity of Alloy 617 indicates that the material is highly strain rate sensitive in the tensile deformation range above about 750°C. This suggests that the concept of a hot tensile curve as a bounding case on the isochronous stress-strain diagrams is problematic. The impact of strain rate on the hot tensile curves is examined and it is concluded that incorporating such a curve is only meaningful if a single tensile strain rate (typically the ASTM standard rate of 0.5%/min) is arbitrarily defined. Current experimentally determined creep data are compared to isochronous stress-strain curves proposed previously by the German programs in the 1980s and by the 1990 draft ASME Code Case. Variability in how well the experimental data are represented by the proposed design curves that suggests further analysis is necessary prior to completing a new draft Code Case.
A summary of the accomplishments, milestones, financial summary, project performance and issues facing the Oxide Production Program for the end of April 2014 is presented in this Executive Summary. PF-4 and ARIES operations remain paused in order to resolve issues and perform an extent-of-condition review for Criticality Safety and Conduct of Operations. On April 28th, Jeff Yarbrough issued a memorandum updating status on the “pause” of PF-4 programmatic operations. He reported that 150 of the procedures that control programmatic operations in PF-4 have successfully resumed. These 150 procedures represent about ½ of the total that will enable us to meet FY14 programmatic commitments. No formal letter of direction will be provided to LANS via the NNSA Field Office contracting officer to address the direction relative to shipments, analytical chemistry sampling and guidance for financial adjustments in FY14 and FY15. LANS will provide a revised plan based on the funding profile included in the President’s Budget Request for the remainder of FY14 and FY15 to include revised FY14 milestones and proposed FY15 milestones. The ARIES Pit Cutter project met the NNSA milestone (Complete Design of Simple Pit Disassembly Equipment [ARIES Pit Cutter] – Due 8/15/14) by completing the fabrication design on 4/3/14, well in advance of the 8/15/14 due date. The project team has started procurement of long-lead items in preparation of the prototype fabrication and cold testing. The fabrication is scheduled to complete in FY14 and the cold testing will begin in FY14 and complete in early FY15. LANL and PNNL met on April 14 to discuss the path forward for PNNL criticality support to the program. The consensus was that PNNL resources can be effectively applied to the generation of detailed CSEs to support the program independent of the current ESS strategy. FY15 program planning is underway. The team is working on planning/drafting the work packages and developing the schedule under the assumption that operations will begin October 1, 2014. A planning schedule has been finalized and the team is meeting regularly to discuss details at the work package and WBS levels. The FY15 PMP is also in the process of being updated. The goal is to have all the drafts reviewed/approved and finalized prior to the start of the FY. MQ personnel have completed ADPSM Management Assessment: Evaluate Quality Roles and Responsibility Agreements between ADPSM and LANL Technical Support Organizations (PFITS 2011-1943). Two findings and four opportunities for improvement were reported. The robotic lathe controller upgrade has entered the construction phase of the project. The controller cabling IWD pre-job meeting was completed. The initial robotic lathe functional test was completed. The existing controller power supply disconnect is scheduled for early May. The NDA Cage project team received notification on 3/28/14 to restart the NDA cage project. Since then the team have been busy with activities that include assigning a planner, getting the construction work package ready for construction and getting the USQ completed. The electrical services installation for the new welding skid for the Inner Can Welder was completed. A gas bottle rack, used to support welding operations, will be installed in early May. The Outer Can Welder design is complete and the IWD is in development to execute construction. Installation materials have been ordered. Work continues on the MD-2 preliminary design. The project engineer is performing a technical evaluation of the Project Management Plan that was submitted by the design agent in response to the LANL Request for Proposal. If all requirements have been captured, LANL procurement will develop a contract and give the design agent notice to proceed on the preliminary design. Estimated completion is scheduled for September 2014. The design for the TA-55 Bldg. 314 warehouse was issued at 90% complete and has begun the last review process. A constructability review is scheduled for early May. The Project Manager has been working with the LANL estimating group to get the class 3 estimate for review. The Oxide Production Cage Upgrade project has kicked-off efforts on the 30% design. Requirements documentation has been drafted. The floor space was walked-down with other project managers and stakeholders and the boundaries were agreed upon.