COMPARISON OF ESCHERICHIA COLI, TOTAL COLIFORM, AND FECAL COLIFORM POPULATIONS AS INDICATORS OF WASTEWATER TREATMENT EFFICIENCY

EPA Science Inventory

Escherichia coli, total coliform, and fecal coliform data were collected from two wastewater treatment facilities, a subsurface constructed wetlands, and the receiving stream. Results are presented from individual wastewater treatment process streams, final effluent and river sit...

Analysis of microbiological contamination in mixed pressed ham and cooked sausage in Korea.

PubMed

Park, Myoung-Su; Wang, Jun; Park, Joong-Hyun; Forghani, Fereidoun; Moon, Jin-San; Oh, Deog-Hwan

2014-03-01

The objective of this study was to investigate the microbial contamination levels (aerobic bacteria plate count [APC], coliforms, Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes) in mixed pressed ham and cooked sausage. A total of 180 samples were collected from factories with and without hazard analysis critical control point (HACCP) systems at four steps: after chopping (AC), after mixing (AM), cooling after the first heating process, and cooling after the second heating process. For ham, APCs and coliform and E. coli counts increased when ingredients were added to the meat at the AC step. Final product APC was 1.63 to 1.85 log CFU/g, and coliforms and E. coli were not detected. S. aureus and L. monocytogenes were found in nine (15.0%) and six (10.0%) samples, respectively, but only at the AC and AM steps and not in the final product. Sausage results were similar to those for ham. The final product APC was 1.52 to 3.85 log CFU/g, and coliforms and E. coli were not detected. S. aureus and L. monocytogenes were found in 29 (24.2%) and 25 (20.8%) samples at the AC and AM steps, respectively, but not in the final product. These results indicate that the temperature and time of the first and second heating are of extreme importance to ensure the microbiological safety of the final product regardless of whether a HACCP system is in place. Microorganism contamination must be monitored regularly and regulations regarding sanitization during processing should be improved. Education regarding employee personal hygiene, environmental hygiene, prevention of cross-contamination, ingredient control, and step-by-step process control is needed to reduce the risk of food poisoning.

Application of DNA hybridization techniques in the assessment of diarrheal disease among refugess in Thailand. [Shigella; Escherichia coli; Campylobacter; Cryptosporidium

DOE Office of Scientific and Technical Information (OSTI.GOV)

Taylor, D.N.; Echeverria, P.; Pitarangsi, C.

1988-01-01

The epidemiology and etiology of acute diarrheal disease were determined in a Hmong refugee camp on the Thai-Laotian border from April 11 to May 14, 1985. DNA hybridization techniques were used to detect Shigella species, enteroinvasive Escherichia coli, and enterotoxigenic E. coli. A monoclonal enzyme-linked immunosorbent assay was used to detect rotavirus, and standard microbiology was used to detect other enteropathogens. The age-specific diarrheal disease rates were 47 episodes per month per 1000 children less than five years old and 113 episodes per month per 1000 children less than one year old. Rotavirus, enterotoxigenic E. coli, Campylobacter, and Cryptosporidium weremore » the predominant pathogens in children less than two years old. The DNA probe hybridized with 94% of 31 specimens identified as enterotoxigenic E. coli by the standard assays and with none of the specimens in which the standard assays were negative. The probe for Shigella and enteroinvasive E. coli hybridized in eight of 10 stools that contained Shigella and four of 314 stools from which Shigella and enteroinvasive E. coli were not isolated. The use of DNA probes allows specimens to be collected in remote areas with a minimum amount of equipment and technical expertise so that they can be easily transported to a central laboratory for further processing.« less

Henry Taube and Coordination Chemistry

Science.gov Websites

Shifts Caused by Cr++ in Aqueous Solutions, DOE Technical Report, 1962 Reactions of Solvated Ions Final Report, DOE Technical Report, 1962 Isotopic Discrimination of Some Solutes in Liquid Ammonia, DOE Technical Report, 1966 Final Technical Report of Research, DOE Technical Report, 1972 Top Additional Web

George A. Olah, Carbocation and Hydrocarbon Chemistry

Science.gov Websites

. Final Technical Report. [HF:BF{sub 2}/H{sub 2}] , DOE Technical Report, 1980 Superacid Catalyzed Coal Conversion Chemistry. 1st and 2nd Quarterly Technical Progress Reports, September 1, 1983-March 30, 1984 , DOE Technical Report, 1984 Superacid Catalyzed Coal Conversion Chemistry. Final Technical Report

The Potential Application and Risks Associated With the Use of Predatory Bacteria as a Biocontrol Agent Against Wound Infections

DTIC Science & Technology

2013-10-01

were isolated and their ability to prey on S . maltophilia (Table-1 and 2) or S . epidermidis (Table-3 and 4) was examined. All experiments were...bacteria ( S . maltophilia or S . epidermidis ) and the host bacteria E. coli strain WM3064, a diaminopimelic acid (DAP) auxotroph. The specific E. coli...times, in each cycle the fraction of the host E. coli was reduced. Finally, Bdellovibrio cells were isolated and their ability to prey on S

77 FR 46306 - Fluxapyroxad; Pesticide Tolerances Technical Amendment

Federal Register 2010, 2011, 2012, 2013, 2014

2012-08-03

...; Pesticide Tolerances Technical Amendment AGENCY: Environmental Protection Agency (EPA). ACTION: Final rule; technical amendment. SUMMARY: EPA issued a final rule in the Federal Register of May 14, 2012, concerning.... Inadvertently, the terminology for the oilseed crop group and for dried plums was incorrect. This technical...

The Technical and Biological Reproducibility of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) Based Typing: Employment of Bioinformatics in a Multicenter Study

PubMed Central

Oberle, Michael; Wohlwend, Nadia; Jonas, Daniel; Maurer, Florian P.; Jost, Geraldine; Tschudin-Sutter, Sarah; Vranckx, Katleen; Egli, Adrian

2016-01-01

Background The technical, biological, and inter-center reproducibility of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI TOF MS) typing data has not yet been explored. The aim of this study is to compare typing data from multiple centers employing bioinformatics using bacterial strains from two past outbreaks and non-related strains. Material/Methods Participants received twelve extended spectrum betalactamase-producing E. coli isolates and followed the same standard operating procedure (SOP) including a full-protein extraction protocol. All laboratories provided visually read spectra via flexAnalysis (Bruker, Germany). Raw data from each laboratory allowed calculating the technical and biological reproducibility between centers using BioNumerics (Applied Maths NV, Belgium). Results Technical and biological reproducibility ranged between 96.8–99.4% and 47.6–94.4%, respectively. The inter-center reproducibility showed a comparable clustering among identical isolates. Principal component analysis indicated a higher tendency to cluster within the same center. Therefore, we used a discriminant analysis, which completely separated the clusters. Next, we defined a reference center and performed a statistical analysis to identify specific peaks to identify the outbreak clusters. Finally, we used a classifier algorithm and a linear support vector machine on the determined peaks as classifier. A validation showed that within the set of the reference center, the identification of the cluster was 100% correct with a large contrast between the score with the correct cluster and the next best scoring cluster. Conclusions Based on the sufficient technical and biological reproducibility of MALDI-TOF MS based spectra, detection of specific clusters is possible from spectra obtained from different centers. However, we believe that a shared SOP and a bioinformatics approach are required to make the analysis robust and reliable. PMID:27798637

Antibiotic resistance patterns in E. coli and Salmonella isolates recovered from commercially available compost

USDA-ARS?s Scientific Manuscript database

Compost is used by both conventional and organic farming practices as an eco-friendly means to enhance soil properties and to reduce fertilizer inputs. Inadequate composting may lead to residual human pathogens (e.g. Escherichia coli O157:H7 and Salmonella spp.) in the final product. Since compost...

Antibiotic resistance patterns in E. coli and Salmonella isolates recovered from commercially vailable compost

USDA-ARS?s Scientific Manuscript database

Introduction: Compost is used by both conventional and organic farming practices as an eco-friendly means to enhance soil properties and to reduce fertilizer inputs. Inadequate composting may lead to residual human pathogens (e.g. Escherichia coli O157:H7 and Salmonella spp.) in the final compost ...

Antibiotic resistance patterns in E. coli and Salmonella isolates recovered from commercially available compost

USDA-ARS?s Scientific Manuscript database

Introduction: Compost is used by both conventional and organic farming practices as an eco-friendly means to enhance soil properties and to reduce fertilizer inputs. Inadequate composting may lead to residual human pathogens (e.g. Escherichia coli O157:H7 and Salmonella spp.) in the final compost ...

Development of a biosensor protein bullet as a fluorescent method for fast detection of Escherichia coli in drinking water.

PubMed

Gutiérrez-Del-Río, Ignacio; Marín, Laura; Fernández, Javier; Álvarez San Millán, María; Ferrero, Francisco Javier; Valledor, Marta; Campo, Juan Carlos; Cobián, Natalia; Méndez, Ignacio; Lombó, Felipe

2018-01-01

Drinking water can be exposed to different biological contaminants from the source, through the pipelines, until reaching the final consumer or industry. Some of these are pathogenic bacteria and viruses which may cause important gastrointestinal or systemic diseases. The microbiological quality of drinking water relies mainly in monitoring three indicator bacteria of faecal origin, Escherichia coli, Enterococcus faecalis and Clostridium perfringens, which serve as early sentinels of potential health hazards for the population. Here we describe the analysis of three chimeric fluorescent protein bullets as biosensor candidates for fast detection of E. coli in drinking water. Two of the chimeric proteins (based on GFP-hadrurin and GFP-pb5 chimera proteins) failed with respect to specificity and/or sensitivity, but the GFP-colS4 chimera protein was able to carry out specific detection of E. coli in drinking water samples in a procedure encompassing about 8 min for final result and this biosensor protein was able to detect in a linear way between 20 and 103 CFU of this bacterium. Below 20 CFU, the system cannot differentiate presence or absence of the target bacterium. The fluorescence in this biosensor system is provided by the GFP subunit of the chimeric protein, which, in the case of the better performing sensor bullet, GFP-colS4 chimera, is covalently bound to a flexible peptide bridge and to a bacteriocin binding specifically to E. coli cells. Once bound to the target bacteria, the excitation step with 395 nm LED light causes emission of fluorescence from the GFP domain, which is amplified in a photomultiplier tube, and finally this signal is converted into an output voltage which can be associated with a CFU value and these data distributed along mobile phone networks, for example. This method, and the portable fluorimeter which has been developed for it, may contribute to reduce the analysis time for detecting E. coli presence in drinking water.

Development of a biosensor protein bullet as a fluorescent method for fast detection of Escherichia coli in drinking water

PubMed Central

Gutiérrez-del-Río, Ignacio; Marín, Laura; Fernández, Javier; Álvarez San Millán, María; Ferrero, Francisco Javier; Valledor, Marta; Campo, Juan Carlos; Cobián, Natalia; Méndez, Ignacio

2018-01-01

Drinking water can be exposed to different biological contaminants from the source, through the pipelines, until reaching the final consumer or industry. Some of these are pathogenic bacteria and viruses which may cause important gastrointestinal or systemic diseases. The microbiological quality of drinking water relies mainly in monitoring three indicator bacteria of faecal origin, Escherichia coli, Enterococcus faecalis and Clostridium perfringens, which serve as early sentinels of potential health hazards for the population. Here we describe the analysis of three chimeric fluorescent protein bullets as biosensor candidates for fast detection of E. coli in drinking water. Two of the chimeric proteins (based on GFP-hadrurin and GFP-pb5 chimera proteins) failed with respect to specificity and/or sensitivity, but the GFP-colS4 chimera protein was able to carry out specific detection of E. coli in drinking water samples in a procedure encompassing about 8 min for final result and this biosensor protein was able to detect in a linear way between 20 and 103 CFU of this bacterium. Below 20 CFU, the system cannot differentiate presence or absence of the target bacterium. The fluorescence in this biosensor system is provided by the GFP subunit of the chimeric protein, which, in the case of the better performing sensor bullet, GFP-colS4 chimera, is covalently bound to a flexible peptide bridge and to a bacteriocin binding specifically to E. coli cells. Once bound to the target bacteria, the excitation step with 395 nm LED light causes emission of fluorescence from the GFP domain, which is amplified in a photomultiplier tube, and finally this signal is converted into an output voltage which can be associated with a CFU value and these data distributed along mobile phone networks, for example. This method, and the portable fluorimeter which has been developed for it, may contribute to reduce the analysis time for detecting E. coli presence in drinking water. PMID:29304041

A comparative approach to recombinantly produce the plant enzyme horseradish peroxidase in Escherichia coli.

PubMed

Gundinger, Thomas; Spadiut, Oliver

2017-04-20

Horseradish peroxidase (HRP) is used in various biotechnological and medical applications. Since its isolation from plant provides several disadvantages, the bacterium Escherichia coli was tested as recombinant expression host in former studies. However, neither production from refolded inclusion bodies nor active enzyme expression in the periplasm exceeded final titres of 10mg per litre cultivation broth. Thus, the traditional way of production of HRP from plant still prevails. In this study, we revisited the recombinant production of HRP in E. coli and investigated and compared both strategies, (a) the production of HRP as inclusion bodies (IBs) and subsequent refolding and (b) the production of active HRP in the periplasm. In fact, we were able to produce HRP in E. coli either way. We obtained a refolding yield of 10% from IBs giving a final titre of 100mgL -1 cultivation broth, and were able to produce 48mg active HRP per litre cultivation broth in the periplasm. In terms of biochemical properties, soluble HRP showed a highly reduced catalytic activity and stability which probably results from the fusion partner DsbA used in this study. Refolded HRP showed similar substrate affinity, an 11-fold reduced catalytic efficiency and 2-fold reduced thermal stability compared to plant HRP. In conclusion, we developed a toolbox for HRP engineering and production. We propose to engineer HRP by directed evolution or semi-rational protein design, express HRP in the periplasm of E. coli allowing straight forward screening for improved variants, and finally produce these variants as IB in high amounts, which are then refolded. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

Beverages: bottled water. Final rule.

PubMed

2009-05-29

The Food and Drug Administration (FDA) is amending its bottled water regulations to require that bottled water manufacturers test source water for total coliform, as is required for finished bottled water products, and to require, if any coliform organisms are detected in source water, that bottled water manufacturers determine whether any of the coliform organisms are Escherichia coli (E. coli), an indicator of fecal contamination. FDA also is amending its bottled water regulations to require, if any coliform organisms are detected in finished bottled water products, that bottled water manufacturers determine whether any of the coliform organisms are E. coli. FDA also is amending the adulteration provision of the bottled water standard to reflect the possibility of adulteration caused by the presence of filth. Bottled water containing E. coli will be considered adulterated, and source water containing E. coli will not be considered to be of a safe, sanitary quality and will be prohibited from use in the production of bottled water. FDA is also amending its bottled water regulations to require that, before a bottler can use source water from a source that has tested positive for E. coli, the bottler must take appropriate measures to rectify or eliminate the cause of E. coli contamination of that source, and that the bottler must keep records of such actions. Existing regulatory provisions require bottled water manufacturers to keep records of new testing required by this rule. This final rule will ensure that FDA's standards for the minimum quality of bottled water, as affected by fecal contamination, will be no less protective of the public health than those set by the Environmental Protection Agency (EPA) for public drinking water.

32 CFR 37.895 - How is the final performance report to be sent to the Defense Technical Information Center?

Code of Federal Regulations, 2014 CFR

2014-07-01

... How is the final performance report to be sent to the Defense Technical Information Center? (a... 32 National Defense 1 2014-07-01 2014-07-01 false How is the final performance report to be sent to the Defense Technical Information Center? 37.895 Section 37.895 National Defense Department of...

32 CFR 37.895 - How is the final performance report to be sent to the Defense Technical Information Center?

Code of Federal Regulations, 2011 CFR

2011-07-01

... How is the final performance report to be sent to the Defense Technical Information Center? (a... 32 National Defense 1 2011-07-01 2011-07-01 false How is the final performance report to be sent to the Defense Technical Information Center? 37.895 Section 37.895 National Defense Department of...

32 CFR 37.895 - How is the final performance report to be sent to the Defense Technical Information Center?

Code of Federal Regulations, 2013 CFR

2013-07-01

... How is the final performance report to be sent to the Defense Technical Information Center? (a... 32 National Defense 1 2013-07-01 2013-07-01 false How is the final performance report to be sent to the Defense Technical Information Center? 37.895 Section 37.895 National Defense Department of...

32 CFR 37.895 - How is the final performance report to be sent to the Defense Technical Information Center?

Code of Federal Regulations, 2012 CFR

2012-07-01

... 32 National Defense 1 2012-07-01 2012-07-01 false How is the final performance report to be sent to the Defense Technical Information Center? 37.895 Section 37.895 National Defense Department of... How is the final performance report to be sent to the Defense Technical Information Center? (a...

Modern space/time geostatistics using river distances: data integration of turbidity and E. coli measurements to assess fecal contamination along the Raritan River in New Jersey.

PubMed

Money, Eric S; Carter, Gail P; Serre, Marc L

2009-05-15

Escherichia coli (E. coli) is a widely used indicator of fecal contamination in water bodies. External contact and subsequent ingestion of bacteria coming from fecal contamination can lead to harmful health effects. Since E. coli data are sometimes limited, the objective of this study is to use secondary information in the form of turbidity to improve the assessment of E. coli at unmonitored locations. We obtained all E. coli and turbidity monitoring data available from existing monitoring networks for the 2000-2006 time period for the Raritan River Basin, New Jersey. Using collocated measurements, we developed a predictive model of E. coli from turbidity data. Using this model, soft data are constructed for E. coli given turbidity measurements at 739 space/time locations where only turbidity was measured. Finally, the Bayesian Maximum Entropy (BME) method of modern space/time geostatistics was used for the data integration of monitored and predicted E. coli data to produce maps showing E. coli concentration estimated daily across the river basin. The addition of soft data in conjunction with the use of river distances reduced estimation error by about 30%. Furthermore, based on these maps, up to 35% of river miles in the Raritan Basin had a probability of E coli impairment greater than 90% on the most polluted day of the study period.

New bactericidal UV light sources: excilamps

NASA Astrophysics Data System (ADS)

Sosnin, Edward A.; Lavrent'eva, Larisa V.; Erofeev, Mikhail V.; Masterova, Yana V.; Kuznetzova, Eugenia N.; Tarasenko, Victor F.

2004-05-01

A reliable bactericidal effect on Escherichia coli cells irradiation by excilamps has been established. Both on primary and secondary irradiation there exists the reciprocally proportional dependence between irradiation doze (or exposure time) and survived cells number. The microorganisms survived after primary irradiation are shown to have not changed sensitivity to excilamps irradiation. The best results have been obtained during XeBr-excilamp irradiation. Owing to their technical parameters, the excilamps are promising systems for UV-sterilization. Comparison of capacitive discharge excilamp characteristics with other conventional UV light sources in presented. A comparative study on UV doze effect of a barrier discharge KrCl-excilamp (λ = 222 nm) on Staphylococcus aureus and Escherichia coli inactivation was carried out. KrCl-excilamp emission power is 65 W, and emitting area is 0.1 m2. It has been demonstrated that Staphylococcus aureus cell sensitivity to UV radiation at this wavelength is higher than that of Escherichia coli.

76 FR 18624 - Research, Technical Assistance and Training Programs: Notice of Final Circular

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-04

... to FTA Circular 6100.1D, Research and Technical Assistance Training Program: Application Instructions... DEPARTMENT OF TRANSPORTATION Federal Transit Administration Research, Technical Assistance and Training Programs: Notice of Final Circular AGENCY: Federal Transit Administration (FTA), DOT. ACTION...

COMPARISON OF THE RECOVERIES OF ESCHERICHIA COLI AND TOTAL COLIFORMS FROM DRINKING WATER BY THE MI AGAR METHOD AND THE U.S. ENVIRONMENTAL PROTECTION AGENCY-APPROVED MEMBRANE FILTER METHOD

EPA Science Inventory

Drinking water regulations under the Final Coliform Rule require that total coliform-positive drinking water samples be examined for the presence of Escherichia coli or fecal coliforms. The current U.S. Environmental Protection Agency-approved membrane filter (MF) method for E. c...

Effects of field storage method on E. coli concentrations measured in storm water runoff.

PubMed

Harmel, Daren; Wagner, Kevin; Martin, Emily; Smith, Doug; Wanjugi, Pauline; Gentry, Terry; Gregory, Lucas; Hendon, Tina

2016-03-01

Storm water runoff is increasingly assessed for fecal indicator organisms (e.g., Escherichia coli, E. coli) and its impact on contact recreation. Concurrently, use of autosamplers along with logistic, economic, technical, and personnel barriers is challenging conventional protocols for sample holding times and storage conditions in the field. A common holding time limit for E. coli is 8 h with a 10 °C storage temperature, but several research studies support longer hold time thresholds. The use of autosamplers to collect E. coli water samples has received little field research attention; thus, this study was implemented to compare refrigerated and unrefrigerated autosamplers and evaluate potential E. coli concentration differences due to field storage temperature (storms with holding times ≤24 h) and due to field storage time and temperature (storms >24 h). Data from 85 runoff events on four diverse watersheds showed that field storage times and temperatures had minor effects on mean and median E. coli concentrations. Graphs and error values did, however, indicate a weak tendency for higher concentrations in the refrigerated samplers, but it is unknown to what extent differing die-off and/or regrowth rates, heterogeneity in concentrations within samples, and laboratory analysis uncertainty contributed to the results. The minimal differences in measured E. coli concentrations cast doubt on the need for utilizing the rigid conventional protocols for field holding time and storage temperature. This is not to say that proper quality assurance and quality control is not important but to emphasize the need to consider the balance between data quality and practical constraints related to logistics, funding, travel time, and autosampler use in storm water studies.

Microbiological quality and safety assessment of lettuce production in Brazil.

PubMed

Ceuppens, Siele; Hessel, Claudia Titze; de Quadros Rodrigues, Rochele; Bartz, Sabrina; Tondo, Eduardo César; Uyttendaele, Mieke

2014-07-02

The microbiological quality and safety of lettuce during primary production in Brazil were determined by enumeration of hygiene indicators Escherichia coli, coliforms and enterococci and detection of enteric pathogens Salmonella and E. coli O157:H7 in organic fertilizers, soil, irrigation water, lettuce crops, harvest boxes and worker's hands taken from six different lettuce farms throughout the crop growth cycle. Generic E. coli was a suitable indicator for the presence of Salmonella and E. coli O157:H7, while coliforms and enterococci were not. Few pathogens were detected: 5 salmonellae and 2 E. coli O157:H7 from 260 samples, of which only one was lettuce and the others were manure, soil and water. Most (5/7) pathogens were isolated from the same farm and all were from organic production. Statistical analysis revealed the following environmental and agro-technical risk factors for increased microbial load and pathogen prevalence in lettuce production: high temperature, flooding of lettuce fields, application of contaminated organic fertilizer, irrigation with water of inferior quality and large distances between the field and toilets. Control of the composting process of organic fertilizers and the irrigation water quality appear most crucial to improve and/or maintain the microbiological quality and safety during the primary production of lettuce. Copyright © 2014 Elsevier B.V. All rights reserved.

Disinfection of Escherichia coli bacteria using hybrid method of ozonation and hydrodynamic cavitation with orifice plate

NASA Astrophysics Data System (ADS)

Karamah, Eva F.; Ghaudenson, Rioneli; Amalia, Fitri; Bismo, Setijo

2017-11-01

This research aims to evaluate the performance of hybrid method of ozonation and hydrodynamic cavitation with orifice plate on E.coli bacteria disinfection. In this research, ozone dose, circulation flowrate, and disinfection method were varied. Ozone was produced by commercial ozonator with ozone dose of 64.83 mg/hour, 108.18 mg/hour, and 135.04 mg/hour. Meanwhile, hydrodynamic cavitation was generated by an orifice plate. The disinfection method compared in this research were: hydrodynamic cavitation, ozonation, and the combination of both. The best result on each method was achieved on the 60th minutes and with a circulation flowrate of 7 L/min. The hybrid method attained final concentration of 0 CFU/mL from the initial concentration of 2.10 × 105 CFU/mL. The ozonation method attained final concentration of 0 CFU/mL from the initial concentration of 1.32 × 105 CFU/mL. Cavitation method gives the least disinfection with final concentration of 5.20 × 104 CFU/mL from the initial concentration of 2.17 × 105 CFU/mL. In conclusion, hybrid method gives a faster and better disinfection of E.coli than each method on its own.

Minnesota Deaf-Blind Technical Assistance Project. Final Report.

ERIC Educational Resources Information Center

Kloos, Eric

This final report describes activities and accomplishments of the 3-year federally supported Minnesota Deaf-Blind Technical Assistance Project. The project provided training and technical assistance, information sharing, and support services to families of children with deaf-blindness. Activities and accomplishments included: collaboration with…

75 FR 56857 - Pilot, Flight Instructor, and Pilot School Certification

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-17

... of part 141. Discussion of Technical Amendment Section 141.5(d) establishes the quality of training... Certification AGENCY: Federal Aviation Administration, DOT. ACTION: Final rule; technical amendment. SUMMARY: The Federal Aviation Administration (FAA) is making minor technical changes to a final rule published...

O157:H7 and O104:H4 Vero/Shiga toxin-producing Escherichia coli outbreaks: respective role of cattle and humans

PubMed Central

2012-01-01

An enteroaggregative Verotoxin (Vtx)-producing Escherichia coli strain of serotype O104:H4 has recently been associated with an outbreak of haemolytic-uremic syndrome and bloody diarrhoea in humans mainly in Germany, but also in 14 other European countries, USA and Canada. This O104:H4 E. coli strain has often been described as an enterohaemorrhagic E. coli (EHEC), i.e. a Vtx-producing E. coli with attaching and effacing properties. Although both EHEC and the German O104:H4 E. coli strains indeed produce Vtx, they nevertheless differ in several other virulence traits, as well as in epidemiological characteristics. For instance, the primary sources and vehicles of typical EHEC infections in humans are ruminants, whereas no animal reservoir has been identified for enteroaggregative E. coli (EAggEC). The present article is introduced by a brief overview of the main characteristics of Vtx-producing E. coli and EAggEC. Thereafter, the O104:H4 E. coli outbreak is compared to typical EHEC outbreaks and the virulence factors and host specificity of EHEC and EAggEC are discussed. Finally, a renewed nomenclature of Vtx-producing E. coli is proposed to avoid more confusion in communication during future outbreaks and to replace the acronym EHEC that only refers to a clinical condition. PMID:22330148

77 FR 29247 - Federal Motor Vehicle Safety Standards; Occupant Crash Protection

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-17

...). ACTION: Final rule; technical amendments. SUMMARY: This final rule makes technical amendments to Federal... advanced air bag requirements. As written now, the general warning label requirements contain an explicit... equipment requirements for restraint systems. This document makes technical amendments to several of the...

Escherichia coli Contamination across Multiple Environmental Compartments (Soil, Hands, Drinking Water, and Handwashing Water) in Urban Harare: Correlations and Risk Factors.

PubMed

Navab-Daneshmand, Tala; Friedrich, Max N D; Gächter, Marja; Montealegre, Maria Camila; Mlambo, Linn S; Nhiwatiwa, Tamuka; Mosler, Hans-Joachim; Julian, Timothy R

2018-03-01

Escherichia coli pathotypes (i.e., enteropathogenic and enterotoxigenic) have been identified among the pathogens most responsible for moderate-to-severe diarrhea in low- and middle-income countries (LMICs). Pathogenic E. coli are transmitted from infected human or animal feces to new susceptible hosts via environmental reservoirs such as hands, water, and soil. Commensal E. coli , which includes nonpathogenic E. coli strains, are widely used as fecal bacteria indicator, with their presence associated with increased likelihood of enteric pathogens and/or diarrheal disease. In this study, we investigated E. coli contamination in environmental reservoirs within households ( N = 142) in high-population density communities of Harare, Zimbabwe. We further assessed the interconnectedness of the environmental compartments by investigating associations between, and household-level risk factors for, E. coli contamination. From the data we collected, the source and risk factors for E. coli contamination are not readily apparent. One notable exception is the presence of running tap water on the household plot, which is associated with significantly less E. coli contamination of drinking water, handwashing water, and hands after handwashing. In addition, E. coli levels on hands after washing are significantly associated with handwashing water contamination, hand contamination before washing, and diarrhea incidence. Finally, we observed that animal ownership increases E. coli contamination in soil, and E. coli in soil are correlated with contamination on hands before washing. This study highlights the complexity of E. coli contamination in household environments within LMICs. More, larger, studies are needed to better identify sources and exposure pathways of E. coli -and enteric pathogens generally-to identify effective interventions.

Pathogenic Escherichia coli producing Extended-Spectrum β-Lactamases isolated from surface water and wastewater.

PubMed

Franz, Eelco; Veenman, Christiaan; van Hoek, Angela H A M; de Roda Husman, Ana; Blaak, Hetty

2015-09-24

To assess public health risks from environmental exposure to Extended-Spectrum β-Lactamases (ESBL)-producing bacteria, it is necessary to have insight in the proportion of relative harmless commensal variants and potentially pathogenic ones (which may directly cause disease). In the current study, 170 ESBL-producing E. coli from Dutch wastewater (n = 82) and surface water (n = 88) were characterized with respect to ESBL-genotype, phylogenetic group, resistance phenotype and virulence markers associated with enteroaggregative E. coli (EAEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), extraintesinal E. coli (ExPEC), and Shiga toxin-producing E. coli (STEC). Overall, 17.1% of all ESBL-producing E. coli were suspected pathogenic variants. Suspected ExPECs constituted 8.8% of all ESBL-producing variants and 8.3% were potential gastrointestinal pathogens (4.1% EAEC, 1.8% EPEC, 1.2% EIEC, 1.2% ETEC, no STEC). Suspected pathogens were significantly associated with ESBL-genotype CTX-M-15 (X(2) = 14.7, P < 0.001) and phylogenetic group B2 (X(2) = 23.5, P < 0.001). Finally, 84% of the pathogenic ESBL-producing E. coli isolates were resistant to three or more different classes of antibiotics. In conclusion, this study demonstrates that the aquatic environment is a potential reservoir of E. coli variants that combine ESBL-genes, a high level of multi-drug resistance and virulence factors, and therewith pose a health risk to humans upon exposure.

Modern Space/Time Geostatistics using River Distances: Data Integration of Turbidity and E.coli Measurements to Assess Fecal Contamination Along the Raritan River in New Jersey

PubMed Central

Money, Eric S.; Carter, Gail P.; Serre, Marc L.

2009-01-01

Escherichia coli (E.coli) is a widely used indicator of fecal contamination in water bodies. External contact and subsequent ingestion of bacteria coming from fecal contamination can lead to harmful health effects. Since E.coli data are sometimes limited, the objective of this study is to use secondary information in the form of turbidity to improve the assessment of E.coli at un-monitored locations. We obtained all E.coli and turbidity monitoring data available from existing monitoring networks for the 2000 – 2006 time period for the Raritan River Basin, New Jersey. Using collocated measurements we developed a predictive model of E.coli from turbidity data. Using this model, soft data are constructed for E.coli given turbidity measurements at 739 space/time locations where only turbidity was measured. Finally, the Bayesian Maximum Entropy (BME) method of modern space/time geostatistics was used for the data integration of monitored and predicted E.coli data to produce maps showing E.coli concentration estimated daily across the river basin. The addition of soft data in conjunction with the use of river distances reduced estimation error by about 30%. Furthermore, based on these maps, up to 35% of river miles in the Raritan Basin had a probability of E.coli impairment greater than 90% on the most polluted day of the study period. PMID:19544881

The Glyphosate-Based Herbicide Roundup Does not Elevate Genome-Wide Mutagenesis of Escherichia coli.

PubMed

Tincher, Clayton; Long, Hongan; Behringer, Megan; Walker, Noah; Lynch, Michael

2017-10-05

Mutations induced by pollutants may promote pathogen evolution, for example by accelerating mutations conferring antibiotic resistance. Generally, evaluating the genome-wide mutagenic effects of long-term sublethal pollutant exposure at single-nucleotide resolution is extremely difficult. To overcome this technical barrier, we use the mutation accumulation/whole-genome sequencing (MA/WGS) method as a mutagenicity test, to quantitatively evaluate genome-wide mutagenesis of Escherichia coli after long-term exposure to a wide gradient of the glyphosate-based herbicide (GBH) Roundup Concentrate Plus. The genome-wide mutation rate decreases as GBH concentration increases, suggesting that even long-term GBH exposure does not compromise the genome stability of bacteria. Copyright © 2017 Tincher et al.

Tolerance development in Listeria monocytogenes-Escherichia coli dual-species biofilms after sublethal exposures to pronase-benzalkonium chloride combined treatments.

PubMed

Rodríguez-López, Pedro; Cabo, Marta López

2017-10-01

This study was designed to assess the effects that sublethal exposures to pronase (PRN) and benzalkonium chloride (BAC) combined treatments have on Listeria monocytogenes-Escherichia coli dual-species biofilms grown on stainless steel in terms of tolerance development (TD) to these compounds. Additionally, fluorescence microscopy was used to observe the changes of the biofilm structure. PRN-BAC exposure was carried out using three different approaches and TD was evaluated treating biofilms with a final 100 μg/ml PRN followed by 50 μg/ml BAC combined treatment. Results showed that exposure to PRN-BAC significantly decreased the number of adhered L. monocytogenes (P < 0.05), while E. coli counts remained generally unaltered. It was also demonstrated that the incorporation of recovery periods during sublethal exposures increased the tolerance of both species of the mixed biofilm to the final PRN-BAC treatment. Moreover, control biofilms became more resistant to PRN-BAC if longer incubation periods were used. Regardless of the treatment used, log reduction values were generally lower in L. monocytogenes compared to E. coli. Additionally, microscopy images showed an altered morphology produced by sublethal PRN-BAC in exposed L. monocytogenes-E. coli dual-species biofilms compared to control samples. Results also demonstrated that L. monocytogenes-E. coli dual-species biofilms are able to develop tolerance to PRN-BAC combined treatments depending on way they have been previously exposed. Moreover, they suggest that the generation of bacterial tolerance should be included as a parameter for sanitation procedures design. Copyright © 2017 Elsevier Ltd. All rights reserved.

Escherichia coli Contamination across Multiple Environmental Compartments (Soil, Hands, Drinking Water, and Handwashing Water) in Urban Harare: Correlations and Risk Factors

PubMed Central

Navab-Daneshmand, Tala; Friedrich, Max N. D.; Gächter, Marja; Montealegre, Maria Camila; Mlambo, Linn S.; Nhiwatiwa, Tamuka; Mosler, Hans-Joachim; Julian, Timothy R.

2018-01-01

Abstract. Escherichia coli pathotypes (i.e., enteropathogenic and enterotoxigenic) have been identified among the pathogens most responsible for moderate-to-severe diarrhea in low- and middle-income countries (LMICs). Pathogenic E. coli are transmitted from infected human or animal feces to new susceptible hosts via environmental reservoirs such as hands, water, and soil. Commensal E. coli, which includes nonpathogenic E. coli strains, are widely used as fecal bacteria indicator, with their presence associated with increased likelihood of enteric pathogens and/or diarrheal disease. In this study, we investigated E. coli contamination in environmental reservoirs within households (N = 142) in high-population density communities of Harare, Zimbabwe. We further assessed the interconnectedness of the environmental compartments by investigating associations between, and household-level risk factors for, E. coli contamination. From the data we collected, the source and risk factors for E. coli contamination are not readily apparent. One notable exception is the presence of running tap water on the household plot, which is associated with significantly less E. coli contamination of drinking water, handwashing water, and hands after handwashing. In addition, E. coli levels on hands after washing are significantly associated with handwashing water contamination, hand contamination before washing, and diarrhea incidence. Finally, we observed that animal ownership increases E. coli contamination in soil, and E. coli in soil are correlated with contamination on hands before washing. This study highlights the complexity of E. coli contamination in household environments within LMICs. More, larger, studies are needed to better identify sources and exposure pathways of E. coli—and enteric pathogens generally—to identify effective interventions. PMID:29363444

Cranberry extract inhibits in vitro adhesion of F4 and F18+Escherichia coli to pig intestinal epithelium and reduces in vivo excretion of pigs orally challenged with F18+ verotoxigenic E. coli.

PubMed

Coddens, Annelies; Loos, Michaela; Vanrompay, Daisy; Remon, Jean Paul; Cox, Eric

2017-04-01

F4 + E. coli and F18 + E. coli infections are an important threat for pig industry worldwide. Antibiotics are commonly used to treat infected piglets, but the emerging development of resistance against antibiotics raises major concerns. Hence, alternative therapies to prevent pigs from F4 + E. coli and F18 + E. coli infections need to be developed. Since cranberry previously showed anti-adhesive activity against uropathogenic E. coli, we aimed to investigate whether cranberry extract could also inhibit binding of F4 + E. coli and F18 + E. coli to pig intestinal epithelium. Using the in vitro villus adhesion assay, we found that low concentrations of cranberry extract (20μg or 100μg/ml) have strong inhibitory activity on F4 + E. coli (75.3%, S.D.=9.31 or 95.8%, S.D.=2.56, respectively) and F18 + E. coli adherence (100% inhibition). This effect was not due to antimicrobial activity. Moreover, cranberry extract (10mg or 100mg) could also abolish in vivo binding of F4 and F18 fimbriae to the pig intestinal epithelium in ligated loop experiments. Finally, two challenge experiments with F18 + E. coli were performed to address the efficacy of in-feed or water supplemented cranberry extract. No effect could be observed in piglets that received cranberry extract only in feed (1g/kg or 10g/kg). However, supplementation of feed (10g/kg) and drinking water (1g/L) significantly decreased excretion and diarrhea. The decreased infection resulted in a decreased serum antibody response indicating reduced exposure to F18 + E. coli. Copyright © 2017 Elsevier B.V. All rights reserved.

77 FR 30512 - Native American Career and Technical Education Program; Final Waivers and Extension of Project...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-23

... DEPARTMENT OF EDUCATION Native American Career and Technical Education Program; Final Waivers and... American Career and Technical Education Program Catalog of Federal Domestic Assistance (CFDA) Number: 84.101A. SUMMARY: For 60-month projects funded in fiscal year (FY) 2007 under the Native American Career...

Effects of administration of four different doses of Escherichia coli phytase on femur properties of 16-week-old turkeys.

PubMed

Tatara, Marcin R; Krupski, Witold; Kozłowski, Krzysztof; Drażbo, Aleksandra; Jankowski, Jan

2015-03-18

The enzyme phytase is able to initiate the release of phosphates from phytic acid, making it available for absorption within gastrointestinal tract and following utilization. The aim of the study was to determine effects of Escherichia coli phytase administration on morphological, densitometric and mechanical properties of femur in 16-week-old turkeys. One-day-old BUT Big-6 males were assigned to six weight-matched groups. Turkeys receiving diet with standard phosphorus (P) and calcium (Ca) content belonged to the positive control group (Group I). Negative control group (Group II) consisted of birds fed diet with lowered P and Ca content. Turkeys belonging to the remaining groups have received the same diet as group II but enriched with graded levels of Escherichia coli phytase: 125 (Group III), 250 (Group IV), 500 (Group V) and 1000 (Group VI) FTU/kg. At the age of 112 days of life, the final body weights were determined and the turkeys were sacrificed to obtain right femur for analyses. Geometric and densitometric properties of femur were determined using quantitative computed tomography (QCT) technique, while mechanical evaluation was performed in three-point bending test. Phytase administration increased cross-sectional area, second moment of inertia, mean relative wall thickness, cortical bone mineral density and maximum elastic strength decreasing cortical bone area of femur (P < 0.05). Reduced dietary Ca and P content decreased final body weight of turkeys by 6.5% (P = 0.006). The most advantageous effects of Escherichia coli phytase administration on geometric, densitometric and mechanical properties of femur were observed in turkeys receiving 125 and 250 FTU/kg of the diet. Phytase administration at the dosages of 500 and 1000 FTU/kg of the diet improved the final body weight in turkeys. The results obtained in this study indicate a possible practical application of Escherichia coli phytase in turkey feeding to improve skeletal system properties and function.

48 CFR 1852.235-73 - Final Scientific and Technical Reports.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 48 Federal Acquisition Regulations System 6 2010-10-01 2010-10-01 true Final Scientific and Technical Reports. 1852.235-73 Section 1852.235-73 Federal Acquisition Regulations System NATIONAL..., including recommendations and conclusions based on the experience and results obtained. The final report...

Coosa River Storage Annex, Talladega, Alabama Environmental Investigation, Final Management and Resources Utilization Plan

DTIC Science & Technology

1991-02-01

to adequately assess the health and environmental risks associated with the closure and transfer of the Annex forI other use; and 3) identification of...1990); Draft Final Technical Plan, Draft Final Sampling Design Plan and Draft Final Health and Safety Plan, USATHAMA, June 1990. 2.1.2 Draft Final...Final Technical Plan, Sampling Design Plan and Health and Safety Plan) supplied by USATHAMA. The estimate may be revised, with USATHAMA approval, as

High-frequency underwater plasma discharge application in antibacterial activity

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ahmed, M. W.; Choi, S.; Lyakhov, K.

Plasma discharge is a novel disinfection and effectual inactivation approach to treat microorganisms in aqueous systems. Inactivation of Gram-negative Escherichia coli (E. coli) by generating high-frequency, high-voltage, oxygen (O{sub 2}) injected and hydrogen peroxide (H{sub 2}O{sub 2}) added discharge in water was achieved. The effect of H{sub 2}O{sub 2} dose and oxygen injection rate on electrical characteristics of discharge and E. coli disinfection has been reported. Microbial log reduction dependent on H{sub 2}O{sub 2} addition with O{sub 2} injection was observed. The time variation of the inactivation efficiency quantified by the log reduction of the initial E. coli population onmore » the basis of optical density measurement was reported. The analysis of emission spectrum recorded after discharge occurrence illustrated the formation of oxidant species (OH{sup •}, H, and O). Interestingly, the results demonstrated that O{sub 2} injected and H{sub 2}O{sub 2} added, underwater plasma discharge had fabulous impact on the E. coli sterilization. The oxygen injection notably reduced the voltage needed for generating breakdown in flowing water and escalated the power of discharge pulses. No impact of hydrogen peroxide addition on breakdown voltage was observed. A significant role of oxidant species in bacterial inactivation also has been identified. Furthermore the E. coli survivability in plasma treated water with oxygen injection and hydrogen peroxide addition drastically reduced to zero. The time course study also showed that the retardant effect on E. coli colony multiplication in plasma treated water was favorable, observed after long time. High-frequency underwater plasma discharge based biological applications is technically relevant and would act as baseline data for the development of novel antibacterial processing strategies.« less

High-frequency underwater plasma discharge application in antibacterial activity

NASA Astrophysics Data System (ADS)

Ahmed, M. W.; Choi, S.; Lyakhov, K.; Shaislamov, U.; Mongre, R. K.; Jeong, D. K.; Suresh, R.; Lee, H. J.

2017-03-01

Plasma discharge is a novel disinfection and effectual inactivation approach to treat microorganisms in aqueous systems. Inactivation of Gram-negative Escherichia coli ( E. coli) by generating high-frequency, high-voltage, oxygen (O2) injected and hydrogen peroxide (H2O2) added discharge in water was achieved. The effect of H2O2 dose and oxygen injection rate on electrical characteristics of discharge and E. coli disinfection has been reported. Microbial log reduction dependent on H2O2 addition with O2 injection was observed. The time variation of the inactivation efficiency quantified by the log reduction of the initial E. coli population on the basis of optical density measurement was reported. The analysis of emission spectrum recorded after discharge occurrence illustrated the formation of oxidant species (OH•, H, and O). Interestingly, the results demonstrated that O2 injected and H2O2 added, underwater plasma discharge had fabulous impact on the E. coli sterilization. The oxygen injection notably reduced the voltage needed for generating breakdown in flowing water and escalated the power of discharge pulses. No impact of hydrogen peroxide addition on breakdown voltage was observed. A significant role of oxidant species in bacterial inactivation also has been identified. Furthermore the E. coli survivability in plasma treated water with oxygen injection and hydrogen peroxide addition drastically reduced to zero. The time course study also showed that the retardant effect on E. coli colony multiplication in plasma treated water was favorable, observed after long time. High-frequency underwater plasma discharge based biological applications is technically relevant and would act as baseline data for the development of novel antibacterial processing strategies.

Point-of-use water disinfection using ultraviolet and visible light-emitting diodes.

PubMed

Lui, Gough Yumu; Roser, David; Corkish, Richard; Ashbolt, Nicholas J; Stuetz, Richard

2016-05-15

Improvements in point-of-use (POU) drinking water disinfection technologies for remote and regional communities are urgently needed. Conceptually, UV-C light-emitting diodes (LEDs) overcome many drawbacks of low-pressure mercury tube based UV devices, and UV-A or visible light LEDs also show potential. To realistically evaluate the promise of LED disinfection, our study assessed the performance of a model 1.3 L reactor, similar in size to solar disinfection bottles. In all, 12 different commercial or semi-commercial LED arrays (270-740 nm) were compared for their ability to inactivate Escherichia coli K12 ATCC W3110 and Enterococcus faecalis ATCC 19433 over 6h. Five log10 and greater reductions were consistently achieved using the 270, 365, 385 and 405 nm arrays. The output of the 310 nm array was insufficient for useful disinfection while 430 and 455 nm performance was marginal (≈ 4.2 and 2.3-log10s E. coli and E. faecalis over the 6h). No significant disinfection was observed with the 525, 590, 623, 660 and 740 nm arrays. Delays in log-phase inactivation of E. coli were observed, particularly with UV-A wavelengths. The radiation doses required for >3-log10 reduction of E. coli and E. faecalis differed by 10 fold at 270 nm but only 1.5-2.5 fold at 365-455 nm. Action spectra, consistent with the literature, were observed with both indicators. The design process revealed cost and technical constraints pertaining to LED electrical efficiency, availability and lifetime. We concluded that POU LED disinfection using existing LED technology is already technically possible. UV-C LEDs offer speed and energy demand advantages, while UV-A/violet units are safer. Both approaches still require further costing and engineering development. Our study provides data needed for such work. Copyright © 2016 Elsevier B.V. All rights reserved.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kimelman, Aya; Levy, Asaf; Sberro, Hila

In the process of clone-based genome sequencing, initial assemblies frequently contain cloning gaps that can be resolved using cloning-independent methods, but the reason for their occurrence is largely unknown. By analyzing 9,328,693 sequencing clones from 393 microbial genomes we systematically mapped more than 15,000 genes residing in cloning gaps and experimentally showed that their expression products are toxic to the Escherichia coli host. A subset of these toxic sequences was further evaluated through a series of functional assays exploring the mechanisms of their toxicity. Among these genes our assays revealed novel toxins and restriction enzymes, and new classes of smallmore » non-coding toxic RNAs that reproducibly inhibit E. coli growth. Further analyses also revealed abundant, short toxic DNA fragments that were predicted to suppress E. coli growth by interacting with the replication initiator dnaA. Our results show that cloning gaps, once considered the result of technical problems, actually serve as a rich source for the discovery of biotechnologically valuable functions, and suggest new modes of antimicrobial interventions.« less

Outer membrane vesicles displaying engineered glycotopes elicit protective antibodies

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen, Linxiao; Valentine, Jenny L.; Huang, Chung-Jr

The O-antigen polysaccharide (O-PS) component of lipopolysaccharides on the surface of gram-negative bacteria is both a virulence factor and a B-cell antigen. Antibodies elicited by O-PS often confer protection against infection; therefore, O-PS glycoconjugate vaccines have proven useful against a number of different pathogenic bacteria. However, conventional methods for natural extraction or chemical synthesis of O-PS are technically demanding, inefficient, and expensive. In this paper, we describe an alternative methodology for producing glycoconjugate vaccines whereby recombinant O-PS biosynthesis is coordinated with vesiculation in laboratory strains of Escherichia coli to yield glycosylated outer membrane vesicles (glycOMVs) decorated with pathogen-mimetic glycotopes. Usingmore » this approach, glycOMVs corresponding to eight different pathogenic bacteria were generated. For example, expression of a 17-kb O-PS gene cluster from the highly virulent Francisella tularensis subsp. tularensis (type A) strain Schu S4 in hypervesiculating E. coli cells yielded glycOMVs that displayed F. tularensis O-PS. Immunization of BALB/c mice with glycOMVs elicited significant titers of O-PS–specific serum IgG antibodies as well as vaginal and bronchoalveolar IgA antibodies. Importantly, glycOMVs significantly prolonged survival upon subsequent challenge with F. tularensis Schu S4 and provided complete protection against challenge with two different F. tularensis subsp. holarctica (type B) live vaccine strains, thereby demonstrating the vaccine potential of glycOMVs. Finally, given the ease with which recombinant glycotopes can be expressed on OMVs, the strategy described here could be readily adapted for developing vaccines against many other bacterial pathogens.« less

Outer membrane vesicles displaying engineered glycotopes elicit protective antibodies

DOE PAGES

Chen, Linxiao; Valentine, Jenny L.; Huang, Chung-Jr; ...

2016-06-06

The O-antigen polysaccharide (O-PS) component of lipopolysaccharides on the surface of gram-negative bacteria is both a virulence factor and a B-cell antigen. Antibodies elicited by O-PS often confer protection against infection; therefore, O-PS glycoconjugate vaccines have proven useful against a number of different pathogenic bacteria. However, conventional methods for natural extraction or chemical synthesis of O-PS are technically demanding, inefficient, and expensive. In this paper, we describe an alternative methodology for producing glycoconjugate vaccines whereby recombinant O-PS biosynthesis is coordinated with vesiculation in laboratory strains of Escherichia coli to yield glycosylated outer membrane vesicles (glycOMVs) decorated with pathogen-mimetic glycotopes. Usingmore » this approach, glycOMVs corresponding to eight different pathogenic bacteria were generated. For example, expression of a 17-kb O-PS gene cluster from the highly virulent Francisella tularensis subsp. tularensis (type A) strain Schu S4 in hypervesiculating E. coli cells yielded glycOMVs that displayed F. tularensis O-PS. Immunization of BALB/c mice with glycOMVs elicited significant titers of O-PS–specific serum IgG antibodies as well as vaginal and bronchoalveolar IgA antibodies. Importantly, glycOMVs significantly prolonged survival upon subsequent challenge with F. tularensis Schu S4 and provided complete protection against challenge with two different F. tularensis subsp. holarctica (type B) live vaccine strains, thereby demonstrating the vaccine potential of glycOMVs. Finally, given the ease with which recombinant glycotopes can be expressed on OMVs, the strategy described here could be readily adapted for developing vaccines against many other bacterial pathogens.« less

Susceptibility of Escherichia coli isolated from uteri of postpartum dairy cows to antibiotic and environmental bacteriophages. Part I: Isolation and lytic activity estimation of bacteriophages.

PubMed

Bicalho, R C; Santos, T M A; Gilbert, R O; Caixeta, L S; Teixeira, L M; Bicalho, M L S; Machado, V S

2010-01-01

The objective of this study was to isolate bacteriophages from environmental samples of 2 large commercial dairy farms using Escherichia coli isolated from the uteri of postpartum Holstein dairy cows as hosts. A total of 11 bacteriophage preparations were isolated from manure systems of commercial dairy farms and characterized for in vitro antimicrobial activity. In addition, a total of 57 E. coli uterine isolates from 5 dairy cows were phylogenetically grouped by triplex PCR. Each E. coli bacterial host from the uterus was inoculated with their respective bacteriophage preparation at several different multiplicities of infections (MOI) to determine minimum inhibitory MOI. The effect of a single dose (MOI=10(2)) of bacteriophage on the growth curve of all 57 E. coli isolates was assessed using a microplate technique. Furthermore, genetic diversity within and between the different bacteriophage preparations was assessed by bacteriophage purification followed by DNA extraction, restriction, and agarose gel electrophoresis. Phylogenetic grouping based on triplex PCR showed that all isolates of E. coli belonged to phylogroup B1. Bacterial growth was completely inhibited at considerably low MOI, and the effect of a single dose (MOI=10(2)) of bacteriophage preparations on the growth curve of all 57 E. coli isolates showed that all bacteriophage preparations significantly decreased the growth rate of the isolates. Bacteriophage preparation 1230-10 had the greatest antimicrobial activity and completely inhibited the growth of 71.7% (n=57) of the isolates. The combined action of bacteriophage preparations 1230-10, 6375-10, 2540-4, and 6547-2, each at MOI=10(2), had the broadest spectrum of action and completely inhibited the growth (final optical density at 600 nm

77 FR 50589 - Agreements and Memoranda of Understanding Between the Food and Drug Administration and Other...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-08-22

...: Final rule. SUMMARY: This final rule makes technical changes that will update a requirement that many of the written agreements and memoranda of understanding (MOUs) between the Food and Drug Administration.... This final rule, accordingly, eliminates it. We are making these technical changes to conserve Agency...

77 FR 30514 - Native Hawaiian Career and Technical Education Program; Final Waiver and Extension of Project Period

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-23

... DEPARTMENT OF EDUCATION Native Hawaiian Career and Technical Education Program; Final Waiver and.... ACTION: Notice. Overview Information Final Waiver and Extension of Project Period for the Native Hawaiian.... SUMMARY: For 36-month projects funded in fiscal year (FY) 2009 under the Native Hawaiian Career and...

Protection against Escherichia coli K1 infection in newborn rats by antibody to K1 capsular polysaccharide antigen.

PubMed

Bortolussi, R; Ferrier, P

1980-04-01

The protective value of antibody to the K1 capsular polysaccharide antigen of Escherichia coli was investigated in a newborn rat model of E. coli K1 infection. Pregnant rats were immunized intravenously with E. coli, and the agglutinating titer to meningococcal group B polysaccharide, which is identical to K1 polysaccharide, was measured in the serum of rats and their offspring. Convalescent serum from rat mothers showed an increased antibody titer in animals injected twice but not once with E. coli K1. Although no agglutinating antibody was detected in the serum of rat pups, animals suckled by mothers having a meningococcal group B agglutinating titer of 1:8 or greater had reduced infection and mortality rates after intraperitoneal injection with E. coli K1 compared with animals suckled by mothers having a low titer of agglutinating antibody (P less than 0.05). In addition, greater protection could be conferred on rat sucklings by oral supplementation with a horse serum rich in antibody to meningococcal group B polysaccharide, suggesting that antibody was abosorbed from the gastrointestinal tract and by itself could be protective. These studies demonstrated that antibody to the capsular polysaccharide of E. coli K1 altered the severity of E. coli K1 infection. Final clearance of bacteria from the blood appeared to await the maturation of other host defense systems in the newborn rat.

Emphysematous endocarditis caused by AmpC beta-lactamase-producing Escherichia coli: A case report.

PubMed

Kim, Chung-Jong; Yi, Jeong-Eun; Kim, Yookyung; Choi, Hee Jung

2018-02-01

Infective endocarditis (IE) is a life-threatening disease, mostly caused by gram-positive bacteria. Gram-negative bacteria were identified as a causative organism in relatively small number of cases. Although, antibiotic-resistant Escherichia coli is common cause of gram-negative endocarditis, AmpC beta-lactamase (BL)-harboring E coli is very rare cause of IE. Furthermore, emphysematous endocarditis is also a very rare manifestation of E coli infection. We report a case of 80-year-old female patient presenting with dizziness, fever, and altered mental status, who was finally diagnosed with emphysematous endocarditis caused by E coli harboring an AmpC BL gene. Her chest computed tomography revealed air bubbles surrounding the annulus of a mitral valve and a transesophageal echocardiogram revealed a hyperechogenic mass fixed on the posteromedial side of the mitral annulus with 2 eccentric mitral regurgitation jets. Blood cultures grew E coli which harbored the DHA-type AmpC BL. The organism belonged to a B2 phylogenic group, and multilocus sequence typing analyses revealed that the strains were of ST-95. She was treated with meropenem following the resistant profiles, and surgery was recommended by the healthcare professional, but denied by the patient's guardians. She was transferred to another hospital due to a refusal for further treatment. Emphysematous endocarditis is an uncommon complication of E coli bacteremia. Certain phylogenetic groups may be associated with development of E coli endocarditis.

Conditional suicide system of Escherichia coli released into soil that uses the Bacillus subtilis sacB gene.

PubMed Central

Recorbet, G; Robert, C; Givaudan, A; Kudla, B; Normand, P; Faurie, G

1993-01-01

The sacB gene from Bacillus subtilis confers sucrose sensitivity upon gram-negative bacteria. The gene was investigated for use as a potential conditional suicide system for Escherichia coli released into soil. To ensure against the loss of the cell death function encoded under nonselective conditions, the nptI-sacR-B suicide cassette was inserted into the E. coli chromosome by using a circular nonreplicative integration vector. Stability studies yielded no loss of the suicide cassette in the integrated E. coli EL1026 strain. sacB induction in the absence of a selective pressure resulted in a lysis efficiency of up to 99.9%. The microcosm experiments confirmed the ability of the suicide cassette to limit the growth and reduce the survival of E. coli strains released into soil. Sucrose addition to sterile soil resulted in a 10(-3)-fold reduction of the final E. coli population density. sacB induction prevented the proliferation and triggered the rapid disappearance of E. coli from natural soil. Mutation to sucrose tolerance occurred at a frequency of 10(-5), making E. coli EL1026 a potential counterselectable donor strain for gene transfer studies. Specificity and potential adaptability to a wide range of gram-negative bacteria are additional conveniences of this conditional suicide system for the containment and counterselection of engineered microorganisms. PMID:8517732

Conditional suicide system of Escherichia coli released into soil that uses the Bacillus subtilis sacB gene.

PubMed

Recorbet, G; Robert, C; Givaudan, A; Kudla, B; Normand, P; Faurie, G

1993-05-01

The sacB gene from Bacillus subtilis confers sucrose sensitivity upon gram-negative bacteria. The gene was investigated for use as a potential conditional suicide system for Escherichia coli released into soil. To ensure against the loss of the cell death function encoded under nonselective conditions, the nptI-sacR-B suicide cassette was inserted into the E. coli chromosome by using a circular nonreplicative integration vector. Stability studies yielded no loss of the suicide cassette in the integrated E. coli EL1026 strain. sacB induction in the absence of a selective pressure resulted in a lysis efficiency of up to 99.9%. The microcosm experiments confirmed the ability of the suicide cassette to limit the growth and reduce the survival of E. coli strains released into soil. Sucrose addition to sterile soil resulted in a 10(-3)-fold reduction of the final E. coli population density. sacB induction prevented the proliferation and triggered the rapid disappearance of E. coli from natural soil. Mutation to sucrose tolerance occurred at a frequency of 10(-5), making E. coli EL1026 a potential counterselectable donor strain for gene transfer studies. Specificity and potential adaptability to a wide range of gram-negative bacteria are additional conveniences of this conditional suicide system for the containment and counterselection of engineered microorganisms.

Monitoring and modeling to predict Escherichia coli at Presque Isle Beach 2, City of Erie, Erie County, Pennsylvania

USGS Publications Warehouse

Zimmerman, Tammy M.

2006-01-01

The Lake Erie shoreline in Pennsylvania spans nearly 40 miles and is a valuable recreational resource for Erie County. Nearly 7 miles of the Lake Erie shoreline lies within Presque Isle State Park in Erie, Pa. Concentrations of Escherichia coli (E. coli) bacteria at permitted Presque Isle beaches occasionally exceed the single-sample bathing-water standard, resulting in unsafe swimming conditions and closure of the beaches. E. coli concentrations and other water-quality and environmental data collected at Presque Isle Beach 2 during the 2004 and 2005 recreational seasons were used to develop models using tobit regression analyses to predict E. coli concentrations. All variables statistically related to E. coli concentrations were included in the initial regression analyses, and after several iterations, only those explanatory variables that made the models significantly better at predicting E. coli concentrations were included in the final models. Regression models were developed using data from 2004, 2005, and the combined 2-year dataset. Variables in the 2004 model and the combined 2004-2005 model were log10 turbidity, rain weight, wave height (calculated), and wind direction. Variables in the 2005 model were log10 turbidity and wind direction. Explanatory variables not included in the final models were water temperature, streamflow, wind speed, and current speed; model results indicated these variables did not meet significance criteria at the 95-percent confidence level (probabilities were greater than 0.05). The predicted E. coli concentrations produced by the models were used to develop probabilities that concentrations would exceed the single-sample bathing-water standard for E. coli of 235 colonies per 100 milliliters. Analysis of the exceedence probabilities helped determine a threshold probability for each model, chosen such that the correct number of exceedences and nonexceedences was maximized and the number of false positives and false negatives was minimized. Future samples with computed exceedence probabilities higher than the selected threshold probability, as determined by the model, will likely exceed the E. coli standard and a beach advisory or closing may need to be issued; computed exceedence probabilities lower than the threshold probability will likely indicate the standard will not be exceeded. Additional data collected each year can be used to test and possibly improve the model. This study will aid beach managers in more rapidly determining when waters are not safe for recreational use and, subsequently, when to issue beach advisories or closings.

The crystal structure of dihydrodipicolinate synthase from Escherichia coli with bound pyruvate and succinic acid semialdehyde: unambiguous resolution of the stereochemistry of the condensation product.

PubMed

Boughton, Berin A; Dobson, Renwick C J; Hutton, Craig A

2012-08-01

The crystal structure of Escherichia coli dihydrodipicolinate synthase with pyruvate and substrate analogue succinic acid semialdehyde condensed with the active site lysine-161 was solved to a resolution of 2.3 Å. Comparative analysis to a previously reported structure both resolves the configuration at the aldol addition center, where the final addition product clearly displays the (S)-configuration, and the final conformation of the adduct within the active site. Direct comparison to two other crystal structures found in the Protein Data Bank, 1YXC, and 3DU0, demonstrates significant similarity between the active site residues of these structures. Copyright © 2012 Wiley Periodicals, Inc.

Faecal contamination of a municipal drinking water distribution system in association with Campylobacter jejuni infections.

PubMed

Pitkänen, Tarja; Miettinen, Ilkka T; Nakari, Ulla-Maija; Takkinen, Johanna; Nieminen, Kalle; Siitonen, Anja; Kuusi, Markku; Holopainen, Arja; Hänninen, Marja-Liisa

2008-09-01

After heavy rains Campylobacter jejuni together with high counts of Escherichia coli, other coliforms and intestinal enterococci were detected from drinking water of a municipal distribution system in eastern Finland in August 2004. Three patients with a positive C. jejuni finding, who had drunk the contaminated water, were identified and interviewed. The pulsed-field gel electrophoresis (PFGE) genotypes from the patient samples were identical to some of the genotypes isolated from the water of the suspected contamination source. In addition, repetitive DNA element analysis (rep-PCR) revealed identical patterns of E. coli and other coliform isolates along the distribution line. Further on-site technical investigations revealed that one of the two rainwater gutters on the roof of the water storage tower had been in an incorrect position and rainwater had flushed a large amount of faecal material from wild birds into the drinking water. The findings required close co-operation between civil authorities, and application of cultivation and genotyping techniques strongly suggested that the municipal drinking water was the source of the infections. The faecal contamination associated with failures in cleaning and technical management stress the importance of instructions for waterworks personnel to perform maintenance work properly.

Genetic Engineering Strategies for Enhanced Biodiesel Production.

PubMed

Hegde, Krishnamoorthy; Chandra, Niharika; Sarma, Saurabh Jyoti; Brar, Satinder Kaur; Veeranki, Venkata Dasu

2015-07-01

The focus on biodiesel research has shown a tremendous growth over the last few years. Several microbial and plant sources are being explored for the sustainable biodiesel production to replace the petroleum diesel. Conventional methods of biodiesel production have several limitations related to yield and quality, which led to development of new engineering strategies to improve the biodiesel production in plants, and microorganisms. Substantial progress in utilizing algae, yeast, and Escherichia coli for the renewable production of biodiesel feedstock via genetic engineering of fatty acid metabolic pathways has been reported in the past few years. However, in most of the cases, the successful commercialization of such engineering strategies for sustainable biodiesel production is yet to be seen. This paper systematically presents the drawbacks in the conventional methods for biodiesel production and an exhaustive review on the present status of research in genetic engineering strategies for production of biodiesel in plants, and microorganisms. Further, we summarize the technical challenges need to be tackled to make genetic engineering technology economically sustainable. Finally, the need and prospects of genetic engineering technology for the sustainable biodiesel production and the recommendations for the future research are discussed.

2005 v4.3 Technical Support Document

EPA Pesticide Factsheets

Emissions Modeling for the Final Mercury and Air Toxics Standards Technical Support Document describes how updated 2005 NEI, version 2 emissions were processed for air quality modeling in support of the final Mercury and Air Toxics Standards (MATS).

The Center for Space Telemetering and Telecommunications Systems

NASA Technical Reports Server (NTRS)

Horan, S.; DeLeon, P.; Borah, D.; Lyman, R.

2003-01-01

This report comprises the final technical report for the research grant 'Center for Space Telemetering and Telecommunications Systems' sponsored by the National Aeronautics and Space Administration's Goddard Space Flight Center. The grant activities are broken down into the following technology areas: (1) Space Protocol Testing; (2) Autonomous Reconfiguration of Ground Station Receivers; (3) Satellite Cluster Communications; and (4) Bandwidth Efficient Modulation. The grant activity produced a number of technical reports and papers that were communicated to NASA as they were generated. This final report contains the final summary papers or final technical report conclusions for each of the project areas. Additionally, the grant supported students who made progress towards their degrees while working on the research.

Promotion of the Equal Access of Girls and Women to Technical and Vocational Education. Studies on Technical and Vocational Education 7.

ERIC Educational Resources Information Center

United Nations Educational, Scientific, and Cultural Organization, Paris (France).

This monograph includes the final report of the International Expert Meeting on the Promotion of Equal Access of Girls and Women to Technical and Vocational Education (TVE) held in Seoul, Republic of Korea, and country discussion papers. The final report is composed of an introduction that proposes that many Member States require special measures…

Effect of land use and hydrological processes on Escherichia coli concentrations in streams of tropical, humid headwater catchments.

PubMed

Rochelle-Newall, Emma J; Ribolzi, Olivier; Viguier, Marion; Thammahacksa, Chanthamousone; Silvera, Norbert; Latsachack, Keooudone; Dinh, Rinh Pham; Naporn, Piyapong; Sy, Hai Tran; Soulileuth, Bounsamay; Hmaimum, Nikom; Sisouvanh, Pem; Robain, Henri; Janeau, Jean-Louis; Valentin, Christian; Boithias, Laurie; Pierret, Alain

2016-09-08

Lack of access to clean water and adequate sanitation continues to be a major brake on development. Here we present the results of a 12-month investigation into the dynamics of Escherichia coli, a commonly used indicator of faecal contamination in water supplies, in three small, rural catchments in Laos, Thailand and Vietnam. We show that land use and hydrology are major controlling factors of E. coli concentrations in streamwater and that the relative importance of these two factors varies between the dry and wet seasons. In all three catchments, the highest concentrations were observed during the wet season when storm events and overland flow were highest. However, smaller peaks of E. coli concentration were also observed during the dry season. These latter correspond to periods of intense farming activities and small, episodic rain events. Furthermore, vegetation type, through land use and soil surface crusting, combined with mammalian presence play an important role in determining E. coli loads in the streams. Finally, sampling during stormflow revealed the importance of having appropriate sampling protocols if information on maximum contamination levels is required as grab sampling at a fixed time step may miss important peaks in E. coli numbers.

Effect of land use and hydrological processes on Escherichia coli concentrations in streams of tropical, humid headwater catchments

NASA Astrophysics Data System (ADS)

Rochelle-Newall, Emma J.; Ribolzi, Olivier; Viguier, Marion; Thammahacksa, Chanthamousone; Silvera, Norbert; Latsachack, Keooudone; Dinh, Rinh Pham; Naporn, Piyapong; Sy, Hai Tran; Soulileuth, Bounsamay; Hmaimum, Nikom; Sisouvanh, Pem; Robain, Henri; Janeau, Jean-Louis; Valentin, Christian; Boithias, Laurie; Pierret, Alain

2016-09-01

Lack of access to clean water and adequate sanitation continues to be a major brake on development. Here we present the results of a 12-month investigation into the dynamics of Escherichia coli, a commonly used indicator of faecal contamination in water supplies, in three small, rural catchments in Laos, Thailand and Vietnam. We show that land use and hydrology are major controlling factors of E. coli concentrations in streamwater and that the relative importance of these two factors varies between the dry and wet seasons. In all three catchments, the highest concentrations were observed during the wet season when storm events and overland flow were highest. However, smaller peaks of E. coli concentration were also observed during the dry season. These latter correspond to periods of intense farming activities and small, episodic rain events. Furthermore, vegetation type, through land use and soil surface crusting, combined with mammalian presence play an important role in determining E. coli loads in the streams. Finally, sampling during stormflow revealed the importance of having appropriate sampling protocols if information on maximum contamination levels is required as grab sampling at a fixed time step may miss important peaks in E. coli numbers.

Air Quality Modeling Technical Support Document for the Final Cross State Air Pollution Rule Update

EPA Pesticide Factsheets

In this technical support document (TSD) we describe the air quality modeling performed to support the final Cross State Air Pollution Rule for the 2008 ozone National Ambient Air Quality Standards (NAAQS).

Production of long chain alcohols and alkanes upon coexpression of an acyl-ACP reductase and aldehyde-deformylating oxgenase with a bacterial type-I fatty acid synthase in E. coli

DOE PAGES

Coursolle, Dan; Shanklin, John; Lian, Jiazhang; ...

2015-06-23

Microbial long chain alcohols and alkanes are renewable biofuels that could one day replace petroleum-derived fuels. Here we report a novel pathway for high efficiency production of these products in Escherichia coli strain BL21(DE3). We first identified the acyl-ACP reductase/aldehyde deformylase combinations with the highest activity in this strain. Next, we used catalase coexpression to remove toxic byproducts and increase the overall titer. Finally, by introducing the type-I fatty acid synthase from Corynebacterium ammoniagenes, we were able to bypass host regulatory mechanisms of fatty acid synthesis that have thus far hampered efforts to optimize the yield of acyl-ACP-derived products inmore » BL21(DE3). When all these engineering strategies were combined with subsequent optimization of fermentation conditions, we were able to achieve a final titer around 100 mg/L long chain alcohol/alkane products including a 57 mg/L titer of pentadecane, the highest titer reported in E. coli BL21(DE3) to date. The expression of prokaryotic type-I fatty acid synthases offer a unique strategy to produce fatty acid-derived products in E. coli that does not rely exclusively on the endogenous type-II fatty acid synthase system.« less

Microbiological contamination in peanut confectionery processing plants.

PubMed

Carminati, J de A; Amorim Neto, D P; Morishita, K N; Takano, L V; Olivier Bernardi, A; Copetti, M V; do Nascimento, M da S

2016-10-01

In order to investigate Enterobacteriaceae, coliforms, Escherichia coli and Salmonella contamination, a survey was conducted at three peanut confectionery processing companies (A, B and C) in Brazil. Samples of different peanut confectionery products (n = 59), peanut raw material (n = 30), manufacturing environment (n = 116) and workers' hand surfaces (n = 12) were analysed. Salmonella and E. coli were not detected in any final product or raw material analysed. Enterobacteriaceae was isolated from 15% of final products. Coliforms were detected in only one sample. Referring to the raw material, six samples showed contamination by Enterobacteriaceae and three samples by coliforms. For the process environment, 19% and 11% of samples presented Enterobacteriaceae and coliforms. Escherichia coli was detected in 5% of samples, and one of these samples tested positive for Salmonella; this strain was serotyping as S. Heidelberg. All food handlers surveyed in Company C showed Enterobacteriaceae and coliforms on their hands. Escherichia coli was isolated from one food worker's hand. The results showed that the manufacturing environment, including food handlers were considered the main sources for possible contamination of peanut confectionery products. This has been the first study to investigate the occurrence of Salmonella and other Enterobacteriaceae throughout peanut confectionery processing lines. The results might be used to assist risk assessment studies and to establish more effective control measures. © 2016 The Society for Applied Microbiology.

Virulence and antimicrobial resistance of Salmonella spp. and Escherichia coli in the beef jerky production line.

PubMed

Fernandes, Fernanda Pereira; Voloski, Flávia Liége Schütz; Ramires, Tassiana; Haubert, Louise; Reta, Giulia Giugliani; Mondadori, Rafael Gianella; Silva, Wladimir Padilha da; Conceição, Rita de Cássia Dos Santos da; Duval, Eduarda Hallal

2017-05-01

Intense manipulation during beef jerky production increases the possibility of contamination with pathogenic microorganisms. This study evaluated the contamination by thermotolerant coliforms, Escherichia coli and Salmonella spp., on processing surfaces and raw materials during beef jerky production, as well as in the final product. Thermotolerant coliforms were found on all surfaces tested and in the raw material. Escherichia coli was identified in 6.7% of the surface samples, while Salmonella spp. was found in 3.3% of the surface samples and 8.6% of raw material samples. Virulence genes were detected in Salmonella spp. isolates. One Salmonella spp. isolate was resistant to sulfonamide, while one E. coli isolate was multiresistant, including the presence of resistance genes sul2, strA, strB, tetA and tetB. The presence of coliforms demonstrates failings in hygienic-sanitary procedures. The presence of pathogenic microorganisms causing foodborne diseases in the production line indicates persistent contamination in the production plant. Although the drying process applied to beef jerky should guarantee the safety of the final product, the presence of multiresistant pathogenic microorganisms, presenting virulence genes, should be a matter of concern. Because beef jerky is a ready-to-eat product, a failure in the production process may cause such microorganisms to pose a public health risk. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Genotypic and phenotypic profiles of Escherichia coli isolates belonging to clinical sequence type 131 (ST131), clinical non-ST131, and fecal non-ST131 lineages from India.

PubMed

Hussain, Arif; Ranjan, Amit; Nandanwar, Nishant; Babbar, Anshu; Jadhav, Savita; Ahmed, Niyaz

2014-12-01

In view of the epidemiological success of CTX-M-15-producing lineages of Escherichia coli and particularly of sequence type 131 (ST131), it is of significant interest to explore its prevalence in countries such as India and to determine if antibiotic resistance, virulence, metabolic potential, and/or the genetic architecture of the ST131 isolates differ from those of non-ST131 isolates. A collection of 126 E. coli isolates comprising 43 ST131 E. coli, 40 non-ST131 E. coli, and 43 fecal E. coli isolates collected from a tertiary care hospital in India was analyzed. These isolates were subjected to enterobacterial repetitive intergenic consensus (ERIC)-based fingerprinting, O typing, phylogenetic grouping, antibiotic sensitivity testing, and virulence and antimicrobial resistance gene (VAG) detection. Representative isolates from this collection were also analyzed by multilocus sequence typing (MLST), conjugation, metabolic profiling, biofilm production assay, and zebra fish lethality assay. All of the 43 ST131 E. coli isolates were exclusively associated with phylogenetic group B2 (100%), while most of the clinical non-ST131 and stool non-ST131 E. coli isolates were affiliated with the B2 (38%) and A (58%) phylogenetic groups, respectively. Significantly greater proportions of ST131 isolates (58%) than non-ST131 isolates (clinical and stool E. coli isolates, 5% each) were technically identified to be extraintestinal pathogenic E. coli (ExPEC). The clinical ST131, clinical non-ST131, and stool non-ST131 E. coli isolates exhibited high rates of multidrug resistance (95%, 91%, and 91%, respectively), extended-spectrum-β-lactamase (ESBL) production (86%, 83%, and 91%, respectively), and metallo-β-lactamase (MBL) production (28%, 33%, and 0%, respectively). CTX-M-15 was strongly linked with ESBL production in ST131 isolates (93%), whereas CTX-M-15 plus TEM were present in clinical and stool non-ST131 E. coli isolates. Using MLST, we confirmed the presence of two NDM-1-positive ST131 E. coli isolates. The aggregate bioscores (metabolite utilization) for ST131, clinical non-ST131, and stool non-ST131 E. coli isolates were 53%, 52%, and 49%, respectively. The ST131 isolates were moderate biofilm producers and were more highly virulent in zebra fish than non-ST131 isolates. According to ERIC-based fingerprinting, the ST131 strains were more genetically similar, and this was subsequently followed by the genetic similarity of clinical non-ST131 and stool non-ST131 E. coli strains. In conclusion, our data provide novel insights into aspects of the fitness advantage of E. coli lineage ST131 and suggest that a number of factors are likely involved in the worldwide dissemination of and infections due to ST131 E. coli isolates. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

7 CFR 614.7 - Preliminary technical determinations.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 6 2010-01-01 2010-01-01 false Preliminary technical determinations. 614.7 Section... Preliminary technical determinations. (a) A preliminary technical determination becomes final 30 days after... purpose of gathering additional information and discussing the facts relating to the preliminary technical...

Differential Distribution of Type II CRISPR-Cas Systems in Agricultural and Nonagricultural Campylobacter coli and Campylobacter jejuni Isolates Correlates with Lack of Shared Environments

PubMed Central

Pearson, Bruce M.; Louwen, Rogier; van Baarlen, Peter; van Vliet, Arnoud H.M.

2015-01-01

CRISPR (clustered regularly interspaced palindromic repeats)-Cas (CRISPR-associated) systems are sequence-specific adaptive defenses against phages and plasmids which are widespread in prokaryotes. Here we have studied whether phylogenetic relatedness or sharing of environmental niches affects the distribution and dissemination of Type II CRISPR-Cas systems, first in 132 bacterial genomes from 15 phylogenetic classes, ranging from Proteobacteria to Actinobacteria. There was clustering of distinct Type II CRISPR-Cas systems in phylogenetically distinct genera with varying G+C%, which share environmental niches. The distribution of CRISPR-Cas within a genus was studied using a large collection of genome sequences of the closely related Campylobacter species Campylobacter jejuni (N = 3,746) and Campylobacter coli (N = 486). The Cas gene cas9 and CRISPR-repeat are almost universally present in C. jejuni genomes (98.0% positive) but relatively rare in C. coli genomes (9.6% positive). Campylobacter jejuni and agricultural C. coli isolates share the C. jejuni CRISPR-Cas system, which is closely related to, but distinct from the C. coli CRISPR-Cas system found in C. coli isolates from nonagricultural sources. Analysis of the genomic position of CRISPR-Cas insertion suggests that the C. jejuni-type CRISPR-Cas has been transferred to agricultural C. coli. Conversely, the absence of the C. coli-type CRISPR-Cas in agricultural C. coli isolates may be due to these isolates not sharing the same environmental niche, and may be affected by farm hygiene and biosecurity practices in the agricultural sector. Finally, many CRISPR spacer alleles were linked with specific multilocus sequence types, suggesting that these can assist molecular epidemiology applications for C. jejuni and C. coli. PMID:26338188

Repetitive Immunosensor with a Fiber-Optic Device and Antibody-Coated Magnetic Beads for Semi-Continuous Monitoring of Escherichia coli O157:H7

PubMed Central

Taniguchi, Midori; Saito, Hirokazu; Mitsubayashi, Kohji

2017-01-01

A rapid and reproducible fiber-optic immunosensor for Escherichia coli O157:H7 (E. coli O157:H7) was described. The biosensor consisted of a flow cell, an optical fiber with a thin Ni layer, and a PC linked fluorometer. First, the samples with E. coli O157:H7 were incubated with magnetic beads coated with anti-E. coli O157:H7 antibodies and anti-E. coli O157:H7 antibodies labeled cyanine 5 (Cy5) to make sandwich complexes. Then the Cy5-(E. coli O157:H7)-beads were injected into a flow cell and pulled to the magnetized Ni layer on the optical fiber set in the flow cell. An excitation light (λ = 635 nm) was used to illuminate the optical fiber, and the Cy5 florescent molecules facing the optical fiber were exposed to an evanescent wave from the optical fiber. The 670 nm fluorescent light was measured using a photodiode. Finally, the magnetic intensity of the Ni layer was removed and the Cy5-E. coli O157:H7-beads were washed out for the next immunoassay. E. coli O157:H7, diluted with phosphate buffer (PB), was measured from 1 × 105 to 1 × 107 cells/mL. The total time required for an assay was less than 15 min (except for the pretreatment process) and repeating immunoassay on one optical fiber was made possible. PMID:28925937

Repetitive Immunosensor with a Fiber-Optic Device and Antibody-Coated Magnetic Beads for Semi-Continuous Monitoring of Escherichia coli O157:H7.

PubMed

Taniguchi, Midori; Saito, Hirokazu; Mitsubayashi, Kohji

2017-09-19

A rapid and reproducible fiber-optic immunosensor for Escherichia coli O157:H7 ( E. coli O157:H7) was described. The biosensor consisted of a flow cell, an optical fiber with a thin Ni layer, and a PC linked fluorometer. First, the samples with E. coli O157:H7 were incubated with magnetic beads coated with anti- E. coli O157:H7 antibodies and anti- E. coli O157:H7 antibodies labeled cyanine 5 (Cy5) to make sandwich complexes. Then the Cy5-( E. coli O157:H7)-beads were injected into a flow cell and pulled to the magnetized Ni layer on the optical fiber set in the flow cell. An excitation light (λ = 635 nm) was used to illuminate the optical fiber, and the Cy5 florescent molecules facing the optical fiber were exposed to an evanescent wave from the optical fiber. The 670 nm fluorescent light was measured using a photodiode. Finally, the magnetic intensity of the Ni layer was removed and the Cy5- E. coli O157:H7-beads were washed out for the next immunoassay. E. coli O157:H7, diluted with phosphate buffer (PB), was measured from 1 × 10⁵ to 1 × 10⁷ cells/mL. The total time required for an assay was less than 15 min (except for the pretreatment process) and repeating immunoassay on one optical fiber was made possible.

Programs of Study as a State Policy Mandate: A Longitudinal Study of the South Carolina Personal Pathways to Success Initiative. Final Technical Report: Major Findings and Implications

ERIC Educational Resources Information Center

Hammond, Cathy; Drew, Sam F.; Withington, Cairen; Griffith, Cathy; Swiger, Caroline M.; Mobley, Catherine; Sharp, Julia L.; Stringfield, Samuel C.; Stipanovic, Natalie; Daugherty, Lindsay

2013-01-01

This is the final technical report from the National Research Center for Career and Technical Education's (NRCCTE's) five-year longitudinal study of South Carolina's Personal Pathway to Success initiative, which was authorized by the state's Education and Economic Development Act (EEDA) in 2005. NRCCTE-affiliated researchers at the National…

Direct Final Rule: Nonroad Diesel Technical Amendments and Tier 3 Technical Relief Provision

EPA Pesticide Factsheets

Rule making certain technical corrections to the rules establishing emission standards for nonroad diesel engines and amending those rules to provide manufacturers with a production technical relief provision for Tier 3 equipment.

Metabolic engineering of the Stevia rebaudiana ent-kaurene biosynthetic pathway in recombinant Escherichia coli.

PubMed

Kong, Min Kyung; Kang, Hyun-Jun; Kim, Jin Ho; Oh, Soon Hwan; Lee, Pyung Cheon

2015-11-20

The ent-kaurene is a dedicated precursor pool and is responsible for synthesizing natural sweeteners such as steviol glycosides. In this study, to produce ent-kaurene in Escherichia coli, we modularly constructed and expressed two ent-kaurene genes encoding ent-copalyl diphosphate synthase (CPPS) and ent-kaurene synthase (KS) from Stevia rebaudiana known as a typical plant producing steviol glycoside. The CPPS and KS from S. rebaudiana were functionally expressed in a heterologous host E. coli. Furthermore, in order to enhance ent-kaurene production in E. coli, six geranylgeranyl diphosphate synthases (GGPPS) from various microorganisms and eight strains of E. coli as host were compared by measuring ent-kaurene production. The highest ent-kaurene production of approximately 41.1mg/L was demonstrated in E. coli strain MG1655 co-expressing synthetic CPPS-KS module and GGPPS from Rhodobacter sphaeroides. The ent-kaurene production was further increased up to 179.6 mg/L by overexpression of the three key enzymes for isoprenoid precursor, 1-deoxyxylulose-5-phosphate synthase (DXS), farnesyl diphosphate synthase (IspA) and isopentenyl diphosphate isomerase (IDI) from E. coli. Finally, the highest titer of ent-kaurene (578 mg/L) with a specific yield of ent-kaurene of 143.5mg/g dry cell weight was obtained by culturing E. coli strain MG1655 co-expressing the ent-kaurene module, DXS, IDI and IspA in 1L bioreactor containing 20 g/L glycerol. Copyright © 2015 Elsevier B.V. All rights reserved.

Gentamicin: effect on E. coli in space

NASA Technical Reports Server (NTRS)

Kacena, M. A.; Todd, P.

1999-01-01

Previous investigations have shown that liquid bacterial cultures grown in space flight were not killed as effectively by antibiotic treatments as were cultures grown on Earth. However, the cause for the decreased antibiotic effectiveness remains unknown. Possible explanations include modified cell proliferation and modified antibiotic transport in the culture medium. Escherichia coli cultures were grown in space flight (STS-69 and STS-73), with and without gentamicin, on a solid agar substrate thus eliminating fluid effects and reducing the unknowns associated with space-flight bacterial cultures in suspension. This research showed that E. coli cultures grown in flight on agar for 24 to 27 hours experienced a heightened growth compared to simultaneous controls. However, addition of gentamicin to the agar killed the bacteria such that both flight and ground control E. coli samples had similar final cell concentrations. Therefore, while the reported existence of a decrease in antibiotic effectiveness in liquid cultures remains unexplained, these data suggest that gentamicin in space flight was at least as effective as, if not more effective than, on Earth, when E. coli cells were grown on agar.

The Potential Application and Risks Associated With the Use of Predatory Bacteria as a Biocontrol Agent Against Wound Infections

DTIC Science & Technology

2014-09-01

host bacteria ( S . maltophilia or S . epidermidis ) and the host bacteria E. coli strain WM3064, a diaminopimelic acid (DAP) auxotroph. The specific E...repeated 11 times, in each cycle the fraction of the host E. coli was reduced. Finally, Bdellovibrio cells were isolated and their ability to prey on S ...maltophilia (Table-1 and 2) or S . epidermidis (Table-3 and 4) was examined. All experiments were conducted in triplicates. Data represent the average

Systems Metabolic Engineering of Escherichia coli.

PubMed

Choi, Kyeong Rok; Shin, Jae Ho; Cho, Jae Sung; Yang, Dongsoo; Lee, Sang Yup

2016-05-01

Systems metabolic engineering, which recently emerged as metabolic engineering integrated with systems biology, synthetic biology, and evolutionary engineering, allows engineering of microorganisms on a systemic level for the production of valuable chemicals far beyond its native capabilities. Here, we review the strategies for systems metabolic engineering and particularly its applications in Escherichia coli. First, we cover the various tools developed for genetic manipulation in E. coli to increase the production titers of desired chemicals. Next, we detail the strategies for systems metabolic engineering in E. coli, covering the engineering of the native metabolism, the expansion of metabolism with synthetic pathways, and the process engineering aspects undertaken to achieve higher production titers of desired chemicals. Finally, we examine a couple of notable products as case studies produced in E. coli strains developed by systems metabolic engineering. The large portfolio of chemical products successfully produced by engineered E. coli listed here demonstrates the sheer capacity of what can be envisioned and achieved with respect to microbial production of chemicals. Systems metabolic engineering is no longer in its infancy; it is now widely employed and is also positioned to further embrace next-generation interdisciplinary principles and innovation for its upgrade. Systems metabolic engineering will play increasingly important roles in developing industrial strains including E. coli that are capable of efficiently producing natural and nonnatural chemicals and materials from renewable nonfood biomass.

In Vivo and in Vitro Synthesis of Phosphatidylglycerol by an Escherichia coli Cardiolipin Synthase.

PubMed

Li, Chijun; Tan, Brandon K; Zhao, Jinshi; Guan, Ziqiang

2016-11-25

Phosphatidylglycerol (PG) makes up 5-20% of the phospholipids of Escherichia coli and is essential for growth in wild-type cells. PG is synthesized from the dephosphorylation of its immediate precursor, phosphatidylglycerol phosphate (PGP) whose synthase in E. coli is PgsA. Using genetic, biochemical, and highly sensitive mass spectrometric approaches, we identified an alternative mechanism for PG synthesis in E. coli that is PgsA independent. The reaction of synthesis involves the conversion of phosphatidylethanolamine and glycerol into PG and is catalyzed by ClsB, a phospholipase D-type cardiolipin synthase. This enzymatic reaction is demonstrated herein both in vivo and in vitro as well as by using the purified ClsB protein. When the growth medium was supplemented with glycerol, the expression of E. coli ClsB significantly increased PG and cardiolipin levels, with the growth deficiency of pgsA null strain also being complemented under such conditions. Identification of this alternative mechanism for PG synthesis not only expands our knowledge of bacterial anionic phospholipid biosynthesis, but also sheds light on the biochemical functions of the cls gene redundancy in E. coli and other bacteria. Finally, the PGP-independent PG synthesis in E. coli may also have important implications for the understanding of PG biosynthesis in eukaryotes that remains incomplete. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Systems Metabolic Engineering of Escherichia coli.

PubMed

Choi, Kyeong Rok; Shin, Jae Ho; Cho, Jae Sung; Yang, Dongsoo; Lee, Sang Yup

2017-03-01

Systems metabolic engineering, which recently emerged as metabolic engineering integrated with systems biology, synthetic biology, and evolutionary engineering, allows engineering of microorganisms on a systemic level for the production of valuable chemicals far beyond its native capabilities. Here, we review the strategies for systems metabolic engineering and particularly its applications in Escherichia coli. First, we cover the various tools developed for genetic manipulation in E. coli to increase the production titers of desired chemicals. Next, we detail the strategies for systems metabolic engineering in E. coli, covering the engineering of the native metabolism, the expansion of metabolism with synthetic pathways, and the process engineering aspects undertaken to achieve higher production titers of desired chemicals. Finally, we examine a couple of notable products as case studies produced in E. coli strains developed by systems metabolic engineering. The large portfolio of chemical products successfully produced by engineered E. coli listed here demonstrates the sheer capacity of what can be envisioned and achieved with respect to microbial production of chemicals. Systems metabolic engineering is no longer in its infancy; it is now widely employed and is also positioned to further embrace next-generation interdisciplinary principles and innovation for its upgrade. Systems metabolic engineering will play increasingly important roles in developing industrial strains including E. coli that are capable of efficiently producing natural and nonnatural chemicals and materials from renewable nonfood biomass.

Arizona Deafblind Project, 1995-1999. Final Report.

ERIC Educational Resources Information Center

Arizona State School for the Deaf and Blind, Tucson.

This final report describes accomplishments of the four-year federally funded Arizona Deafblind Project which attempted to: (1) identify all deafblind children in Arizona; (2) deliver technical assistance to families; (3) deliver technical assistance to service providers; and (4) enhance community oversight, coordination, and collaboration with…

Collection methods and quality assessment for Esche-richia coli, water quality, and microbial source tracking data within Tumacácori National Historical Park and the upper Santa Cruz River, Arizona, 2015-16

USGS Publications Warehouse

Paretti, Nicholas; Coes, Alissa L.; Kephart, Christopher M.; Mayo, Justine

2018-03-05

Tumacácori National Historical Park protects the culturally important Mission, San José de Tumacácori, while also managing a portion of the ecologically diverse riparian corridor of the Santa Cruz River. This report describes the methods and quality assurance procedures used in the collection of water samples for the analysis of Escherichia coli (E. coli), microbial source tracking markers, suspended sediment, water-quality parameters, turbidity, and the data collection for discharge and stage; the process for data review and approval is also described. Finally, this report provides a quantitative assessment of the quality of the E. coli, microbial source tracking, and suspended sediment data.The data-quality assessment revealed that bias attributed to field and laboratory contamination was minimal, with E. coli detections in only 3 out of 33 field blank samples analyzed. Concentrations in the field blanks were several orders of magnitude lower than environmental concentrations. The microbial source tracking (MST) field blank was below the detection limit for all MST markers analyzed. Laboratory blanks for E. coli at the USGS Arizona Water Science Center and laboratory blanks for MST markers at the USGS Ohio Water Microbiology Laboratory were all below the detection limit. Irreplicate data for E. coli and suspended sediment indicated that bias was not introduced to the data by combining samples collected using discrete sampling methods with samples collected using automatic sampling methods.The split and sequential E. coli replicate data showed consistent analytical variability and a single equation was developed to explain the variability of E. coli concentrations. An additional analysis of analytical variability for E. coli indicated analytical variability around 18 percent relative standard deviation and no trend was observed in the concentration during the processing and analysis of multiple split-replicates. Two replicate samples were collected for MST and individual markers were compared for a base flow and flood sample. For the markers found in common between the two types of samples, the relative standard deviation for the base flow sample was more than 3 times greater than the markers in the flood sample. Sequential suspended sediment replicates had a relative standard deviation of about 1.3 percent, indicating that environmental and analytical variability was minimal.A holding time review and laboratory study analysis supported the extended holding times required for this investigation. Most concentrations for flood and base-flow samples were within the theoretical variability specified in the most probable number approach suggesting that extended hold times did not overly influence the final concentrations reported.

Effects of sublethal concentrations of silver nanoparticles on Escherichia coli and Bacillus subtilis under aerobic and anaerobic conditions.

PubMed

Garuglieri, Elisa; Cattò, Cristina; Villa, Federica; Zanchi, Raffaella; Cappitelli, Francesca

2016-12-16

The present work is aimed at comparing the effects of sublethal concentrations of silver nanoparticles (AgNPs) on the growth kinetic, adhesion ability, oxidative stress, and phenotypic changes of model bacteria (Escherichia coli and Bacillus subtilis) under both aerobic and anaerobic conditions. Growth kinetic tests conducted in 96-well microtiter plates revealed that sublethal concentrations of AgNPs do not affect E. coli growth, whereas 1 μg/ml AgNPs increased B. subtilis growth rate under aerobic conditions. At the same concentration, AgNPs promoted B. subtilis adhesion, while it discouraged E. coli attachment to the surface in the presence of oxygen. As determined by 2,7-dichlorofluorescein-diacetate assays, AgNPs increased the formation of intracellular reactive oxygen species, but not at the highest concentrations, suggesting the activation of scavenging systems. Finally, motility assays revealed that 0.01 and 1 μg/ml AgNPs, respectively, promoted surface movement in E. coli and B. subtilis under aerobic and anaerobic conditions. The results demonstrate that E. coli and B. subtilis react differently from AgNPs over a wide range of sublethal concentrations examined under both aerobic and anaerobic conditions. These findings will help elucidate the behavior and impact of engineered nanoparticles on microbial ecosystems.

Effect of land use and hydrological processes on Escherichia coli concentrations in streams of tropical, humid headwater catchments

PubMed Central

Rochelle-Newall, Emma J.; Ribolzi, Olivier; Viguier, Marion; Thammahacksa, Chanthamousone; Silvera, Norbert; Latsachack, Keooudone; Dinh, Rinh Pham; Naporn, Piyapong; Sy, Hai Tran; Soulileuth, Bounsamay; Hmaimum, Nikom; Sisouvanh, Pem; Robain, Henri; Janeau, Jean-Louis; Valentin, Christian; Boithias, Laurie; Pierret, Alain

2016-01-01

Lack of access to clean water and adequate sanitation continues to be a major brake on development. Here we present the results of a 12-month investigation into the dynamics of Escherichia coli, a commonly used indicator of faecal contamination in water supplies, in three small, rural catchments in Laos, Thailand and Vietnam. We show that land use and hydrology are major controlling factors of E. coli concentrations in streamwater and that the relative importance of these two factors varies between the dry and wet seasons. In all three catchments, the highest concentrations were observed during the wet season when storm events and overland flow were highest. However, smaller peaks of E. coli concentration were also observed during the dry season. These latter correspond to periods of intense farming activities and small, episodic rain events. Furthermore, vegetation type, through land use and soil surface crusting, combined with mammalian presence play an important role in determining E. coli loads in the streams. Finally, sampling during stormflow revealed the importance of having appropriate sampling protocols if information on maximum contamination levels is required as grab sampling at a fixed time step may miss important peaks in E. coli numbers. PMID:27604854

Joint Common Architecture Demonstration (JCA Demo) Final Report

DTIC Science & Technology

2016-07-28

approach for implementing open systems [16], formerly known as the Modular Open Systems Approach (MOSA). OSA is a business and technical strategy to... TECHNICAL REPORT RDMR-AD-16-01 JOINT COMMON ARCHITECTURE DEMONSTRATION (JCA DEMO) FINAL REPORT Scott A. Wigginton... Modular Avionics .......................................................................... 5 E. Model-Based Engineering

Transfusion of stored blood impairs host defenses against Gram-negative pathogens in mice.

PubMed

Prestia, Kevin; Bandyopadhyay, Sheila; Slate, Andrea; Francis, Richard O; Francis, Kevin P; Spitalnik, Steven L; Fidock, David A; Brittenham, Gary M; Hod, Eldad A

2014-11-01

Although human red blood cell (RBC) units may be refrigerator stored for up to 42 days, transfusion of older RBCs acutely delivers a large bolus of iron to mononuclear phagocytes. Similarly, iron dextran circulates in plasma for hours to days and is progressively cleared by mononuclear phagocytes, which return iron to plasma. Finally, malaria infection continuously delivers iron to macrophages by intra- and extravascular hemolysis. Studies suggest that iron administration increases infectious risk. To assess the effects of increased iron availability on susceptibility to infection, we infected mice with model Gram-negative intracellular or extracellular pathogens (Salmonella typhimurium or Escherichia coli, respectively), accompanied by RBC transfusion, iron dextran administration, or malarial coinfection. In our mouse models, transfusion of older RBCs exacerbates infection with both Gram-negative pathogens. Although iron dextran exacerbates E. coli infection to a similar extent as transfusion of corresponding amounts of iron, higher iron doses are required to produce comparable effects with S. typhimurium. Coinfection of mice with Plasmodium yoelii and S. typhimurium produces overwhelming Salmonella sepsis. Finally, treating mice with antibiotics abrogates the enhancing effect on E. coli infection of both older RBC transfusion and iron dextran administration. Transfusion of older RBCs exacerbates Gram-negative infection to a similar extent as malaria coinfection or iron dextran administration. Appropriate antibiotic therapy abrogates the effect of older RBC transfusions on infection with E. coli. Iron delivery to macrophages may be an underappreciated mechanism mediating, at least some, adverse effects of RBC transfusions. © 2014 AABB.

Biodegradation of Aromatic Compounds by Escherichia coli

PubMed Central

Díaz, Eduardo; Ferrández, Abel; Prieto, María A.; García, José L.

2001-01-01

Although Escherichia coli has long been recognized as the best-understood living organism, little was known about its abilities to use aromatic compounds as sole carbon and energy sources. This review gives an extensive overview of the current knowledge of the catabolism of aromatic compounds by E. coli. After giving a general overview of the aromatic compounds that E. coli strains encounter and mineralize in the different habitats that they colonize, we provide an up-to-date status report on the genes and proteins involved in the catabolism of such compounds, namely, several aromatic acids (phenylacetic acid, 3- and 4-hydroxyphenylacetic acid, phenylpropionic acid, 3-hydroxyphenylpropionic acid, and 3-hydroxycinnamic acid) and amines (phenylethylamine, tyramine, and dopamine). Other enzymatic activities acting on aromatic compounds in E. coli are also reviewed and evaluated. The review also reflects the present impact of genomic research and how the analysis of the whole E. coli genome reveals novel aromatic catabolic functions. Moreover, evolutionary considerations derived from sequence comparisons between the aromatic catabolic clusters of E. coli and homologous clusters from an increasing number of bacteria are also discussed. The recent progress in the understanding of the fundamentals that govern the degradation of aromatic compounds in E. coli makes this bacterium a very useful model system to decipher biochemical, genetic, evolutionary, and ecological aspects of the catabolism of such compounds. In the last part of the review, we discuss strategies and concepts to metabolically engineer E. coli to suit specific needs for biodegradation and biotransformation of aromatics and we provide several examples based on selected studies. Finally, conclusions derived from this review may serve as a lead for future research and applications. PMID:11729263

Selection of Single Domain Antibodies from Immune Libraries Displayed on the Surface of E. coli Cells with Two β-Domains of Opposite Topologies

PubMed Central

Martínez-Arteaga, Rocio; Ruano-Gallego, David; Fraile, Sofía; Margolles, Yago; Teira, Xema; Gutierrez, Carlos; Bodelón, Gustavo; Fernández, Luis Ángel

2013-01-01

Screening of antibody (Ab) libraries by direct display on the surface of E. coli cells is hampered by the presence of the outer membrane (OM). In this work we demonstrate that the native β-domains of EhaA autotransporter and intimin, two proteins from enterohemorrhagic E. coli O157:H7 (EHEC) with opposite topologies in the OM, are effective systems for the display of immune libraries of single domain Abs (sdAbs) from camelids (nanobodies or VHH) on the surface of E. coli K-12 cells and for the selection of high affinity sdAbs using magnetic cell sorting (MACS). We analyzed the capacity of EhaA and intimin β-domains to display individual sdAbs and sdAb libraries obtained after immunization with the extracellular domain of the translocated intimin receptor from EHEC (TirMEHEC). We demonstrated that both systems displayed functional sdAbs on the surface of E. coli cells with little proteolysis and cellular toxicity, although E. coli cells displaying sdAbs with the β-domain of intimin showed higher antigen-binding capacity. Both E. coli display libraries were screened for TirMEHEC binding clones by MACS. High affinity binders were selected by both display systems, although more efficiently with the intimin β-domain. The specificity of the selected clones against TirMEHEC was demonstrated by flow cytometry of E. coli cells, along with ELISA and surface plasmon resonance with purified sdAbs. Finally, we employed the E. coli cell display systems to provide an estimation of the affinity of the selected sdAb by flow cytometry analysis under equilibrium conditions. PMID:24086454

Development of a rapid high-efficiency scalable process for acetylated Sus scrofa cationic trypsin production from Escherichia coli inclusion bodies.

PubMed

Zhao, Mingzhi; Wu, Feilin; Xu, Ping

2015-12-01

Trypsin is one of the most important enzymatic tools in proteomics and biopharmaceutical studies. Here, we describe the complete recombinant expression and purification from a trypsinogen expression vector construct. The Sus scrofa cationic trypsin gene with a propeptide sequence was optimized according to Escherichia coli codon-usage bias and chemically synthesized. The gene was inserted into pET-11c plasmid to yield an expression vector. Using high-density E. coli fed-batch fermentation, trypsinogen was expressed in inclusion bodies at 1.47 g/L. The inclusion body was refolded with a high yield of 36%. The purified trypsinogen was then activated to produce trypsin. To address stability problems, the trypsin thus produced was acetylated. The final product was generated upon gel filtration. The final yield of acetylated trypsin was 182 mg/L from a 5-L fermenter. Our acetylated trypsin product demonstrated higher BAEE activity (30,100 BAEE unit/mg) than a commercial product (9500 BAEE unit/mg, Promega). It also demonstrated resistance to autolysis. This is the first report of production of acetylated recombinant trypsin that is stable and suitable for scale-up. Copyright © 2015 Elsevier Inc. All rights reserved.

Hedgehog Protein Cholesterolysis: A New Therapeutic Target for Advanced Prostate Cancer

DTIC Science & Technology

2016-10-01

research group page, with links to publicationshttp://www.binghamton.edu/chemistry/people/callahan/callahan.html s 5. PARTICIPANTS & OTHER COLLABORATING...which changes the side chain by only a methylene group . All four expressed as soluble proteins in E. coli and exhibited strong FRET signals consistent...Acknowledgements We thank members of the Callahan research group for technical assistance and helpful comments on the manuscript. Special thanks to Dr

76 FR 80226 - Technical Amendments

Federal Register 2010, 2011, 2012, 2013, 2014

2011-12-23

... final rule, effective upon publication. Generally, the Administrative Procedure Act (APA) requires a.... Additionally, the APA requires that a final rule must have a delayed effective date of 30 days from the date of... delayed effective date requirement under the APA. 5 U.S.C. 553(d)(3). Again the technical change conforms...

"Type Ia Supernovae: Tools for Studying Dark Energy" Final Technical Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Woosley, Stan; Kasen, Dan

2017-05-10

Final technical report for project "Type Ia Supernovae: Tools for the Study of Dark Energy" awarded jointly to scientists at the University of California, Santa Cruz and Berkeley, for computer modeling, theory and data analysis relevant to the use of Type Ia supernovae as standard candles for cosmology.

76 FR 50202 - National Technical Assistance and Dissemination Center for Children Who Are Deaf-Blind; Final...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-12

... DEPARTMENT OF EDUCATION [CFDA No. 84.326T] National Technical Assistance and Dissemination Center for Children Who Are Deaf-Blind; Final Extension of Project Period and Waiver AGENCY: Office of Special Education Programs, Office of Special Education and Rehabilitative Services, Department of...

TADS Final Evaluation Report, 1980-81. Appendix S.

ERIC Educational Resources Information Center

Suarez, Tanya M.; And Others

The document contains the final report of the Technical Assistance Development System (TADS), a program which provided technical assistance (TA) services to 53 Handicapped Children's Early Education Program (HCEEP) demonstration projects and 13 State Implementation Grants (SIGs). The evaluation report is divided into five sections. Section 1…

DipTest: A litmus test for E. coli detection in water.

PubMed

Gunda, Naga Siva Kumar; Dasgupta, Saumyadeb; Mitra, Sushanta K

2017-01-01

We have developed a new litmus paper test (DipTest) for detecting Escherichia coli (E. coli) in water samples by performing enzymatic reactions directly on the porous paper substrate. The paper strip consists of a long narrow piece of cellulose blotting paper coated with chemoattractant (at bottom edge), wax hydrophobic barrier (at the top edge), and custom formulated chemical reagents (at reaction zone immediately below the wax hydrophobic barrier). When the paper strip is dipped in water, E. coli in the water sample is attracted toward the paper strip due to a chemotaxic mechanism followed by the ascent along the paper strip toward the reaction zone due to a capillary wicking mechanism, and finally the capillary motion is arrested at the top edge of the paper strip by the hydrophobic barrier. The E. coli concentrated at the reaction zone of the paper strip will react with custom formulated chemical reagents to produce a pinkish-red color. Such a color change on the paper strip when dipped into water samples indicates the presence of E. coli contamination in potable water. The performance of the DipTest device has been checked with different known concentrations of E. coli contaminated water samples using different dip and wait times. The DipTest device has also been tested with different interfering bacteria and chemical contaminants. It has been observed that the different interfering contaminants do not have any impact on the DipTest, and it can become a potential solution for screening water samples for E. coli contamination at the point of source.

DipTest: A litmus test for E. coli detection in water

PubMed Central

Gunda, Naga Siva Kumar; Dasgupta, Saumyadeb

2017-01-01

We have developed a new litmus paper test (DipTest) for detecting Escherichia coli (E. coli) in water samples by performing enzymatic reactions directly on the porous paper substrate. The paper strip consists of a long narrow piece of cellulose blotting paper coated with chemoattractant (at bottom edge), wax hydrophobic barrier (at the top edge), and custom formulated chemical reagents (at reaction zone immediately below the wax hydrophobic barrier). When the paper strip is dipped in water, E. coli in the water sample is attracted toward the paper strip due to a chemotaxic mechanism followed by the ascent along the paper strip toward the reaction zone due to a capillary wicking mechanism, and finally the capillary motion is arrested at the top edge of the paper strip by the hydrophobic barrier. The E. coli concentrated at the reaction zone of the paper strip will react with custom formulated chemical reagents to produce a pinkish-red color. Such a color change on the paper strip when dipped into water samples indicates the presence of E. coli contamination in potable water. The performance of the DipTest device has been checked with different known concentrations of E. coli contaminated water samples using different dip and wait times. The DipTest device has also been tested with different interfering bacteria and chemical contaminants. It has been observed that the different interfering contaminants do not have any impact on the DipTest, and it can become a potential solution for screening water samples for E. coli contamination at the point of source. PMID:28877199

Cloning of the Alcaligenes latus polyhydroxyalkanoate biosynthesis genes and use of these genes for enhanced production of Poly(3-hydroxybutyrate) in Escherichia coli.

PubMed

Choi, J I; Lee, S Y; Han, K

1998-12-01

Polyhydroxyalkanoates (PHAs) are microbial polyesters that can be used as completely biodegradable polymers, but the high production cost prevents their use in a wide range of applications. Recombinant Escherichia coli strains harboring the Ralstonia eutropha PHA biosynthesis genes have been reported to have several advantages as PHA producers compared with wild-type PHA-producing bacteria. However, the PHA productivity (amount of PHA produced per unit volume per unit time) obtained with these recombinant E. coli strains has been lower than that obtained with the wild-type bacterium Alcaligenes latus. To endow the potentially superior PHA biosynthetic machinery to E. coli, we cloned the PHA biosynthesis genes from A. latus. The three PHA biosynthesis genes formed an operon with the order PHA synthase, beta-ketothiolase, and reductase genes and were constitutively expressed from the natural promoter in E. coli. Recombinant E. coli strains harboring the A. latus PHA biosynthesis genes accumulated poly(3-hydroxybutyrate) (PHB), a model PHA product, more efficiently than those harboring the R. eutropha genes. With a pH-stat fed-batch culture of recombinant E. coli harboring a stable plasmid containing the A. latus PHA biosynthesis genes, final cell and PHB concentrations of 194.1 and 141.6 g/liter, respectively, were obtained, resulting in a high productivity of 4.63 g of PHB/liter/h. This improvement should allow recombinant E. coli to be used for the production of PHB with a high level of economic competitiveness.

Total Coliform Rule (TCR) Federal Register Notice

EPA Pesticide Factsheets

This document provides the FR notice to 40 CFR Parts 141 and 142 Drinking Water: National Primary Drinking Water Regulations; Total Coliforms (Including Fecal Coliforms and E. Coli); Final Rule (26 pp, 5 M).

Treatment of Mycobacterium intracellulare Infected Mice with Walter Reed Compound H

DTIC Science & Technology

1980-09-25

including Mycobacterium tuberculosis, Mycobacterium leprae , Staphylococcusaureus, Pseudomonas aeruginosa, Escherichia coli, enterococ----ci, Neissr•~n...97 SAD "Treatment of Mycobacterium intracellulare Infected Mice with Walter Reed Compound H" Final Comprehensive Report J. Kenneth McClatchy, Ph.D...REPORT & PERIOD COVERED "TREATMENT OF MYCOBACTERIUM INTRACELLULARE - Final Comprehensive Report INFECTED MICE WITH WALTER REED COMPOUND H"li G

Simulation of Graphene Field-Effect Transistor Biosensors for Bacterial Detection.

PubMed

Wu, Guangfu; Meyyappan, Meyya; Lai, King Wai Chiu

2018-05-25

Foodborne illness is correlated with the existence of infectious pathogens such as bacteria in food and drinking water. Probe-modified graphene field effect transistors (G-FETs) have been shown to be suitable for Escherichia coli ( E. coli ) detection. Here, the G-FETs for bacterial detection are modeled and simulated with COMSOL Multiphysics to understand the operation of the biosensors. The motion of E. coli cells in electrolyte and the surface charge of graphene induced by E. coli are systematically investigated. The comparison between the simulation and experimental data proves the sensing probe size to be a key parameter affecting the surface charge of graphene induced by bacteria. Finally, the relationship among the change in source-drain current (∆ I ds ), graphene-bacteria distance and bacterial concentration is established. The shorter graphene-bacteria distance and higher bacterial concentration give rise to better sensing performance (larger ∆ I ds ) of the G-FETs biosensors. The simulation here could serve as a guideline for the design and optimization of G-FET biosensors for various applications.

Double resonance long period fiber grating for detection of E. coli in trace concentration by choosing a proper bacteriophage

NASA Astrophysics Data System (ADS)

Chiniforooshan, Y.; Celebanska, A.; Janik, M.; Mikulic, P.; Haddad, F.; Perreault, J.; Bock, W. J.

2017-04-01

There is a critical need of a fast, specific and reliable assay for biological species. To address this need, long period fiber gratings (LPFG) among other fiber optic sensors can be used because of their high sensitivity to changes in surrounding medium. In this work we fabricated and used two over-etched LPFGs. One of them was covered with T4 Phage and the other was covered with MS2 phage that both specifically bind with Escherichia coli (E. coli) bacteria. This bacterium is a major cause of the food contaminations and outbreaks. We showed achieving a highest sensitivity region of the LPFG and the way to fine tune to that region by over-etching the grating. Finally, using the highly sensitive LPFG platform we could detect E. coli at concentrations as low as 100 colony forming units (CFU), by covering the LPFG with an optimized bio-functionalization of the fiber surface with MS2 bacteriophage.

A low cost technique for synthesis of gold nanoparticles using microwave heating and its application in signal amplification for detecting Escherichia Coli O157:H7 bacteria

NASA Astrophysics Data System (ADS)

Thanh Ngo, Vo Ke; Giang Nguyen, Dang; Phat Huynh, Trong; Lam, Quang Vinh

2016-09-01

In the present work a low cost technique for preparation of gold nanoparticles (AuNPs) using microwave heating was developed. The effect of different elements (precursor reagents, irradiation time, and microwave radiation power) on the final morphology of AuNPs obtained through the microwave assisted technique has been investigated. The characterization of the samples has been carried out by transmission electron microscopy, UV-vis absorption spectroscopy, Fourier transform infrared spectroscopy, and powder x-ray diffraction. The results showed that to some extent the above-mentioned characterizations influenced the size of synthetized nanoparticles and application of microwave heating has many advantages such as low cost, rapid preparation and highly uniform particles. As an application in quartz crystal microbalance (QCM) immunosensor, AuNPs are conjugated with the Escherichia coli (E.coli) O157:H7 antibodies for signal amplification to detect E.coli O157:H7 bacteria residual in QCM system.

Quantification of loosely associated and tightly associated bacteria on broiler carcass skin using swabbing, stomaching, and grinding methods.

PubMed

Singh, P; Lee, H C; Chin, K B; Ha, S D; Kang, I

2015-12-01

This research was conducted to quantify bacterial populations after swabbing or stomaching, followed by grinding the swabbed or stomached broiler skins. For each of 3 replications, 3 eviscerated broilers were randomly taken from a processing line in a local broiler processing plant. Ten swabs and 10 stomachs per bird were conducted on the left- and the right-side skins (10×7 cm), respectively, which were then finally ground. Results indicated that mesophilic aerobic bacteria (MAB) in the first swabbed sample were significantly lower than those in the first stomached sample (P<0.05), with no difference seen for the remaining sampling times (P>0.05). During 10 swabbings followed by final grinding, 8, 9, and 83% of MAB were detected after the first swabbing, after the second through 10th swabbings, and after final grinding of the skin, respectively. During 10 stomachings followed by the final grinding, 17, 18, and 65% of MAB were detected after the first stomaching, after the second through 10th stomachings, and after final grinding of the skin, respectively. Escherichia coli (E. coli) and coliforms were significantly higher in the first stomaching than those in the first swabbing (P<0.05), with no difference seen between the 2 sampling methods for the rest sampling times (P>0.05). Populations of E. coli and coliforms decreased step-wisely from the highest after grinding to the intermediate after first and second sampling, and to the least after 10th sampling (P<0.05), regardless of swabbing or grinding. In this study, less than 35% of MAB seemed loosely associated in the skin of eviscerated broiler, whereas more than 65% of MAB looked tightly associated, which were not recovered by stomaching or swabbing even 10 times but were recovered by grinding the skin. © 2015 Poultry Science Association Inc.

Characterization of beta-carotene ketolases, CrtW, from marine bacteria by complementation analysis in Escherichia coli.

PubMed

Choi, Seon-kang; Nishida, Yasuhiro; Matsuda, Satoru; Adachi, Kyoko; Kasai, Hiroaki; Peng, Xue; Komemushi, Sadao; Miki, Wataru; Misawa, Norihiko

2005-01-01

A complementation analysis was performed in Escherichia coli to evaluate the efficiency of beta-carotene ketolases (CrtW) from the marine bacteria Brevundimonas sp. SD212, Paracoccus sp. PC1 (Alcaligenes PC-1), and Paracoccus sp. N81106 (Agrobacterium aurantiacum), for astaxanthin production. Each crtW gene was expressed in Escherichia coli synthesizing zeaxanthin due to the presence of plasmid pACCAR25DeltacrtX. Carotenoids that accumulated in the resulting E. coli transformants were examined by chromatographic and spectroscopic analyses. The transformant carrying the Paracoccus sp. PC1 or N81106 crtW gene accumulated high levels of adonixanthin, which is the final astaxanthin precursor for CrtW, and astaxanthin, while the E. coli transformant with crtW from Brevundimonas sp. SD212 did not accumulate any adonixanthin and produced a high level of astaxanthin. These results show efficient conversion by CrtW of Brevundimonas sp. SD212 from adonixanthin to astaxanthin, which is a new-found characteristic of a bacterial CrtW enzyme. The phylogenetic positions between CrtW of the two genera, Brevundimonas and Paracoccus, are distant, although they fall into alpha-Proteobacteria.

Engineering Bacteria to Catabolize the Carbonaceous Component of Sarin: Teaching E. coli to Eat Isopropanol

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brown, Margaret E.; Mukhopadhyay, Aindrila; Keasling, Jay D.

In this paper, we report an engineered strain of Escherichia coli that catabolizes the carbonaceous component of the extremely toxic chemical warfare agent sarin. Enzymatic decomposition of sarin generates isopropanol waste that, with this engineered strain, is then transformed into acetyl-CoA by enzymatic conversion with a key reaction performed by the acetone carboxylase complex (ACX). We engineered the heterologous expression of the ACX complex from Xanthobacter autotrophicus PY2 to match the naturally occurring subunit stoichiometry and purified the recombinant complex from E. coli for biochemical analysis. Incorporating this ACX complex and enzymes from diverse organisms, we introduced an isopropanol degradationmore » pathway in E. coli, optimized induction conditions, and decoupled enzyme expression to probe pathway bottlenecks. Our engineered E. coli consumed 65% of isopropanol compared to no-cell controls and was able to grow on isopropanol as a sole carbon source. Finally, in the process, reconstitution of this large ACX complex (370 kDa) in a system naïve to its structural and mechanistic requirements allowed us to study this otherwise cryptic enzyme in more detail than would have been possible in the less genetically tractable native Xanthobacter system.« less

Solid-state voltammetry-based electrochemical immunosensor for Escherichia coli using graphene oxide-Ag nanoparticle composites as labels.

PubMed

Jiang, Xiaochun; Chen, Kun; Wang, Jing; Shao, Kang; Fu, Tao; Shao, Feng; Lu, Donglian; Liang, Jiangong; Foda, M Frahat; Han, Heyou

2013-06-21

A new electrochemical immunosensor based on solid-state voltammetry was fabricated for the detection of Escherichia coli (E. coli) by using graphene oxide-Ag nanoparticle composites (P-GO-Ag) as labels. To construct the platform, Au nanoparticles (AuNPs) were first self-assembled on an Au electrode surface through cysteamine and served as an effective matrix for antibody (Ab) attachment. Under a sandwich-type immunoassay format, the analyte and the probe (P-GO-Ag-Ab) were successively captured onto the immunosensor. Finally, the bonded AgNPs were detected through a solid-state redox process in 0.2 M of KCl solution. Combining the advantages of the high-loading capability of graphene oxide with promoted electron-transfer rate of AuNPs, this immunosensor produced a 26.92-fold signal enhancement compared with the unamplified protocol. Under the optimal conditions, the immunosensor exhibited a wide linear dependence on the logarithm of the concentration of E. coli ranging from 50 to 1.0 × 10(6) cfu mL(-1) with a detection limit of 10 cfu mL(-1). Moreover, as a practical application, the proposed immunosensor was used to monitor E. coli in lake water with satisfactory results.

78 FR 57264 - Final Waiver and Extension of the Project Period for the Technical Assistance Coordination Center

Federal Register 2010, 2011, 2012, 2013, 2014

2013-09-18

...: 84.326Z.] Final Waiver and Extension of the Project Period for the Technical Assistance Coordination... project period. SUMMARY: The Secretary waives the requirements in the Education Department General Administrative Regulations that generally prohibit project periods exceeding five years and extensions of project...

77 FR 16923 - Agreements and Memoranda of Understanding Between the Food and Drug Administration and Other...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-23

...: Direct final rule. SUMMARY: This direct final rule makes technical changes that will update a requirement that many of our written agreements and memoranda of understanding (MOUs) with other departments..., accordingly, eliminates it. We are making these technical changes to conserve Agency time and resources...

1988 Underground Storage Tanks; Technical Requirements; Final Rule and Underground Storage Tanks Containing Petroleum-Financial Responsibility Requirements and State Program Approval Objective; Final Rule

EPA Pesticide Factsheets

EPA's 1988 regulations concerning USTs are contained in 40 CFR Part 280, 40 CFR Part 281 and 40 CFR Parts 282.50-282.105 and divided into three sections: technical requirements, financial responsibility requirements, and state program approval objectives.

PITTSBURGH TECHNICAL HEALTH TRAINING INSTITUTE DEMONSTRATION PROJECT. FINAL REPORT, VOLUME II.

ERIC Educational Resources Information Center

KISHKUNAS, LOUIS J.

APPENDIXES TO THE "FINAL REPORT," VOLUME I (VT 005 511), ARE INCLUDED--(1) A SCHEMATIC REPRESENTATION OF CURRICULUM DEVELOPMENT, (2) TECHNICAL BEHAVIOR CHECKLISTS, (3) PERFORMANCE INVENTORY FORMS USED IN ON-THE-JOB OBSERVATIONS, (4) REPORT FORM FOR TYPICAL JOB BEHAVIOR OF EMPLOYEE, (5) COOPERATING AREA HEALTH INSTITUTIONS, (6) TABLES OF Z SCORES…

Differential Distribution of Type II CRISPR-Cas Systems in Agricultural and Nonagricultural Campylobacter coli and Campylobacter jejuni Isolates Correlates with Lack of Shared Environments.

PubMed

Pearson, Bruce M; Louwen, Rogier; van Baarlen, Peter; van Vliet, Arnoud H M

2015-09-02

CRISPR (clustered regularly interspaced palindromic repeats)-Cas (CRISPR-associated) systems are sequence-specific adaptive defenses against phages and plasmids which are widespread in prokaryotes. Here we have studied whether phylogenetic relatedness or sharing of environmental niches affects the distribution and dissemination of Type II CRISPR-Cas systems, first in 132 bacterial genomes from 15 phylogenetic classes, ranging from Proteobacteria to Actinobacteria. There was clustering of distinct Type II CRISPR-Cas systems in phylogenetically distinct genera with varying G+C%, which share environmental niches. The distribution of CRISPR-Cas within a genus was studied using a large collection of genome sequences of the closely related Campylobacter species Campylobacter jejuni (N = 3,746) and Campylobacter coli (N = 486). The Cas gene cas9 and CRISPR-repeat are almost universally present in C. jejuni genomes (98.0% positive) but relatively rare in C. coli genomes (9.6% positive). Campylobacter jejuni and agricultural C. coli isolates share the C. jejuni CRISPR-Cas system, which is closely related to, but distinct from the C. coli CRISPR-Cas system found in C. coli isolates from nonagricultural sources. Analysis of the genomic position of CRISPR-Cas insertion suggests that the C. jejuni-type CRISPR-Cas has been transferred to agricultural C. coli. Conversely, the absence of the C. coli-type CRISPR-Cas in agricultural C. coli isolates may be due to these isolates not sharing the same environmental niche, and may be affected by farm hygiene and biosecurity practices in the agricultural sector. Finally, many CRISPR spacer alleles were linked with specific multilocus sequence types, suggesting that these can assist molecular epidemiology applications for C. jejuni and C. coli. © The Author(s) 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

Invention and Writing in Technical Work: Representing the Object.

ERIC Educational Resources Information Center

Winsor, Dorothy A.

1994-01-01

Describes the way invention is relevant to the practice of technical writing. Studies three engineering students engaged in a real-world project. Shows how the students' technical work and invention for the final report were simultaneous activities. Claims that invention for and through writing overlaps with technical invention. (HB)

Project T.E.A.M. (Technical Education Advancement Modules). Final Report.

ERIC Educational Resources Information Center

Greenville Technical Coll., SC.

Project TEAM (Technical Education Advancement Modules), a cooperative demonstration program for high technology training, created an introductory technical training program and a consumer education package emphasizing the benefits of technical training. The curriculum and training focus of the project began with an assessment of employee needs in…

77 FR 39623 - Airworthiness Standards: Aircraft Engines; Technical Amendment

Federal Register 2010, 2011, 2012, 2013, 2014

2012-07-05

...] Airworthiness Standards: Aircraft Engines; Technical Amendment AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Final rule; technical amendment. SUMMARY: This amendment clarifies aircraft engine... from applicants requesting FAA engine type certifications and aftermarket certifications, such as...

A Family of Indoles Regulate Virulence and Shiga Toxin Production in Pathogenic E. coli

PubMed Central

Izrayelit, Yevgeniy; Bhatt, Shantanu; Cartwright, Emily; Wang, Wei; Swimm, Alyson I.; Benian, Guy M.; Schroeder, Frank C.; Kalman, Daniel

2013-01-01

Enteropathogenic Escherichia coli (EPEC), enterohemorrhagic E. coli (EHEC) and enteroaggregative E. coli (EAEC) are intestinal pathogens that cause food and water-borne disease in humans. Using biochemical methods and NMR-based comparative metabolomics in conjunction with the nematode Caenorhabditis elegans, we developed a bioassay to identify secreted small molecules produced by these pathogens. We identified indole, indole-3-carboxaldehyde (ICA), and indole-3-acetic acid (IAA), as factors that only in combination are sufficient to kill C. elegans. Importantly, although lethal to C. elegans, these molecules downregulate several bacterial processes important for pathogenesis in mammals. These include motility, biofilm formation and production of Shiga toxins. Some pathogenic E. coli strains are known to contain a Locus of Enterocyte Effacement (LEE), which encodes virulence factors that cause “attaching and effacing” (A/E) lesions in mammals, including formation of actin pedestals. We found that these indole derivatives also downregulate production of LEE virulence factors and inhibit pedestal formation on mammalian cells. Finally, upon oral administration, ICA inhibited virulence and promoted survival in a lethal mouse infection model. In summary, the C. elegans model in conjunction with metabolomics has facilitated identification of a family of indole derivatives that broadly regulate physiology in E. coli, and virulence in pathogenic strains. These molecules may enable development of new therapeutics that interfere with bacterial small-molecule signaling. PMID:23372726

Glyceraldehyde-3-phosphate dehydrogenase: a universal internal control for Western blots in prokaryotic and eukaryotic cells.

PubMed

Wu, Yonghong; Wu, Min; He, Guowei; Zhang, Xiao; Li, Weiguang; Gao, Yan; Li, Zhihui; Wang, Zhaoyan; Zhang, Chenggang

2012-04-01

In the current study, we examined the expression level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein in a number of organisms and the stability of GAPDH under various conditions. Our results revealed that GAPDH is present in multiple Escherichia coli strains, the yeast strain GS115, Caenorhabditis elegans, rat PC12 cells, and both mouse and rat brain. Furthermore, GAPDH was stably expressed under different concentrations of inducer and at different times of induction in E. coli (BL21) cells and yeast GS115 cells. Stable expression of GAPDH protein was also observed in C.elegans and PC12 cells that were treated with different concentrations of paraquat or sodium sulfite, respectively. In addition, we were able to detect and identify the endogenous gapA protein in E.coli via immunoprecipitation and MALDI-TOF-MS analysis. Endogenous gapA protein and exogenously expressed (subcloned) GAPDH proteins were detected in E. coli BL21 but not for gapC. With the exception of gapC in E. coli, the various isoforms of GAPDH possessed enzymatic activity. Finally, sequence analysis revealed that the GAPDH proteins were 76% identical, with the exception of E. coli gapC. Taken together, our results indicate that GAPDH could be universally used as an internal control for the Western blot analysis of prokaryotic and eukaryotic samples. Crown Copyright © 2012. Published by Elsevier Inc. All rights reserved.

Systems metabolic engineering of Escherichia coli for production of the antitumor drugs violacein and deoxyviolacein.

PubMed

Rodrigues, André L; Trachtmann, Nathalie; Becker, Judith; Lohanatha, Ananta F; Blotenberg, Jana; Bolten, Christoph J; Korneli, Claudia; de Souza Lima, André O; Porto, Luismar M; Sprenger, Georg A; Wittmann, Christoph

2013-11-01

Violacein and deoxyviolacein are interesting therapeutics against pathogenic bacteria and viruses as well as tumor cells. In the present work, systems-wide metabolic engineering was applied to target Escherichia coli, a widely accepted recombinant host in pharmaceutical biotechnology, for production of these high-value products. The basic producer, E. coli dVio-1, that expressed the vioABCE cluster from Chromobacterium violaceum under control of the inducible araC system, accumulated 180 mg L(-1) of deoxyviolacein. Targeted intracellular metabolite analysis then identified bottlenecks in tryptophan supporting pathways, the major product building block. This was used for comprehensive engineering of serine, chorismate and tryptophan biosynthesis and the non-oxidative pentose-phosphate pathway. The final strain, E. coli dVio-6, accumulated 320 mg L(-1) deoxyviolacein in shake flask cultures. The created chassis of a high-flux tryptophan pathway was complemented by genomic integration of the vioD gene of Janthinobacterium lividum, which enabled exclusive production of violacein. In a fed-batch process, the resulting producer E. coli Vio-4 accumulated 710 mg L(-1) of the desired product. With straightforward broth extraction and subsequent crystallization, violacein could be obtained with 99.8% purity. This demonstrates the potential of E. coli as a platform for production of tryptophan based therapeutics. Copyright © 2013 Elsevier Inc. All rights reserved.

Field Evidence Supporting Conventional Onion Curing Practices as a Strategy To Mitigate Escherichia coli Contamination from Irrigation Water.

PubMed

Wright, Daniel; Feibert, Erik; Reitz, Stuart; Shock, Clint; Waite-Cusic, Joy

2018-03-01

The Produce Safety Rule of the U.S. Food Safety Modernization Act includes restrictions on the use of agricultural water of poor microbiological quality. Mitigation options for poor water quality include the application of an irrigation-to-harvest interval of <4 days; however, dry bulb onion production includes an extended irrigation-to-harvest interval (<30 days). This study evaluated conventional curing practices for mitigating Escherichia coli contamination in a field setting. Well water inoculated with rifampin-resistant E. coli (1, 2, or 3 log CFU/mL) was applied to onion fields (randomized block design; n = 5) via drip tape on the final day of irrigation. Onions remained undisturbed for 7 days and were then lifted to the surface to cure for an additional 21 days before harvest. Water, onions, and soil were tested for presence of rifampin-resistant E. coli. One day after irrigation, 13.3% of onions (20 of 150) receiving the poorest quality water (3 log CFU/mL) tested positive for E. coli; this prevalence was reduced to 4% (6 of 150 onions) after 7 days. Regardless of inoculum level, E. coli was not detected on any onions beyond 15 days postirrigation. These results support conventional dry bulb onion curing practices as an effective strategy to mitigate microbiological concerns associated with poor quality irrigation water.

The impact of fecal sample processing on prevalence estimates for antibiotic-resistant Escherichia coli.

PubMed

Omulo, Sylvia; Lofgren, Eric T; Mugoh, Maina; Alando, Moshe; Obiya, Joshua; Kipyegon, Korir; Kikwai, Gilbert; Gumbi, Wilson; Kariuki, Samuel; Call, Douglas R

2017-05-01

Investigators often rely on studies of Escherichia coli to characterize the burden of antibiotic resistance in a clinical or community setting. To determine if prevalence estimates for antibiotic resistance are sensitive to sample handling and interpretive criteria, we collected presumptive E. coli isolates (24 or 95 per stool sample) from a community in an urban informal settlement in Kenya. Isolates were tested for susceptibility to nine antibiotics using agar breakpoint assays and results were analyzed using generalized linear mixed models. We observed a <3-fold difference between prevalence estimates based on freshly isolated bacteria when compared to isolates collected from unprocessed fecal samples or fecal slurries that had been stored at 4°C for up to 7days. No time-dependence was evident (P>0.1). Prevalence estimates did not differ for five distinct E. coli colony morphologies on MacConkey agar plates (P>0.2). Successive re-plating of samples for up to five consecutive days had little to no impact on prevalence estimates. Finally, culturing E. coli under different conditions (with 5% CO 2 or micro-aerobic) did not affect estimates of prevalence. For the conditions tested in these experiments, minor modifications in sample processing protocols are unlikely to bias estimates of the prevalence of antibiotic-resistance for fecal E. coli. Copyright © 2017 Elsevier B.V. All rights reserved.

Identifying Key Proteins in Hg Methylation Pathways of Desulfovibrio by Global Proteomics, Final Technical Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Summers, Anne O.; Miller, Susan M.; Wall, Judy

2016-06-18

Elemental mercury, Hg(0) is a contaminant at many DOE sites, especially at Oak Ridge National Laboratory (ORNL) where the spread of spilled Hg and its effects on microbial populations have been monitored for decades. To explore the microbial interactions with Hg, we have devised a global proteomic approach capable of directly detecting Hg-adducts of proteins. This technique developed in the facultative anaerobe, Escherichia coli, allows us to identify the proteins most vulnerable to acute exposure to organomercurials phenyl- and ethyl-mercury (as surrogates for the highly neurotoxic methyl-Hg) (Polacco, et al, 2011). We have found >300 such proteins in all metabolicmore » functional groups and cellular compartments; most are highly conserved and can serve as markers for acute Hg exposure (Zink, et al. 2016, in preparation). We have also discovered that acute Hg exposure severely disrupts thiol, iron and redox homeostases, and electrolyte balance (LaVoie, et al., 2015) Thus, we proposed to bring these techniques to bear on the central problem of identifying the cellular proteins involved in bacterial uptake and methylation of mercury and its release from the cell.« less

Antibiotic resistant bacteria in urban sewage: Role of full-scale wastewater treatment plants on environmental spreading.

PubMed

Turolla, A; Cattaneo, M; Marazzi, F; Mezzanotte, V; Antonelli, M

2018-01-01

The presence of antibiotic resistant bacteria (ARB) in wastewater was investigated and the role of wastewater treatment plants (WWTPs) in promoting or limiting antibiotic resistance was assessed. Escherichia coli (E. coli) and total heterotrophic bacteria (THB) resistance to ampicillin, chloramphenicol and tetracycline was monitored in three WWTPs located in Milan urban area (Italy), differing among them for the operating parameters of biological process, for the disinfection processes (based on sodium hypochlorite, UV radiation, peracetic acid) and for the discharge limits to be met. Wastewater was collected from three sampling points along the treatment sequence (WWTP influent, effluent from sand filtration, WWTP effluent). Antibiotic resistance to ampicillin was observed both for E. coli and for THB. Ampicillin resistant bacteria in the WWTP influents were 20-47% of E. coli and 16-25% of THB counts. A limited resistance to chloramphenicol was observed only for E. coli, while neither for E. coli nor for THB tetracycline resistance was observed. The biological treatment and sand filtration led to a decrease in the maximum percentage of ampicillin-resistant bacteria (20-29% for E. coli, 11-21% for THB). However, the conventionally adopted parameters did not seem adequate to support an interpretation of WWTP role in ARB spread. Peracetic acid was effective in selectively acting on antibiotic resistant THB, unlike UV radiation and sodium hypochlorite. The low counts of E. coli in WWTP final effluents in case of agricultural reuse did not allow to compare the effect of the different disinfection processes on antibiotic resistance. Copyright © 2017 Elsevier Ltd. All rights reserved.

7 CFR 652.36 - Appeal of decertification decisions.

Code of Federal Regulations, 2010 CFR

2010-01-01

... technical service provider's written appeal, the Chief or his designee, will make a final determination, in... CONSERVATION SERVICE, DEPARTMENT OF AGRICULTURE SUPPORT ACTIVITIES TECHNICAL SERVICE PROVIDER ASSISTANCE... of the State Conservationist's decertification determination, the technical service provider may...

Development of a high yielding E. coli periplasmic expression system for the production of humanized Fab' fragments.

PubMed

Ellis, Mark; Patel, Pareshkumar; Edon, Marjory; Ramage, Walter; Dickinson, Robert; Humphreys, David P

2017-01-01

Humanized Fab' fragments may be produced in the periplasm of Escherichia coli but can be subject to degradation by host cell proteases. In order to increase Fab' yield and reduce proteolysis we developed periplasmic protease deficient strains of E. coli. These strains lacked the protease activity of Tsp, protease III and DegP. High cell density fermentations indicated Tsp deficient strains increased productivity two fold but this increase was accompanied by premature cell lysis soon after the induction of Fab' expression. To overcome the reduction in cell viability we introduced suppressor mutations into the spr gene. The mutations partially restored the wild type phenotype of the cells. Furthermore, we coexpressed a range of periplasmic chaperone proteins with the Fab', DsbC had the most significant impact, increasing humanized Fab' production during high cell density fermentation. When DsbC coexpression was combined with a Tsp deficient spr strain we observed an increase in yield and essentially restored "wild type" cell viability. We achieved a final periplasmic yield of over 2.4g/L (final cell density OD 600 105), 40 h post Fab' induction with minimal cell lysis.The data suggests that proteolysis, periplasm integrity, protein folding and disulphide bond formation are all potential limiting steps in the production of Fab' fragments in the periplasm of E. coli. In this body of work, we have addressed these limiting steps by utilizing stabilized protease deficient strains and chaperone coexpression. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:212-220, 2017. © 2016 American Institute of Chemical Engineers.

Epidemiological studies on Escherichia coli O157:H7 in Egyptian sheep.

PubMed

Kamel, Mohammed; Abo El-Hassan, Diea G; El-Sayed, Amr

2015-08-01

In the present work, the epidemiological role of apparently healthy sheep in transmission of Escherichia coli O157:H7 in different seasons was investigated. Fecal samples (convenience sampling) of apparently healthy farmed sheep (three farms, n = 70) and from 15 wandering flocks fed on city wastes (n = 80) in the Giza governorate were examined. The samples were collected in spring under mild weather conditions and during hot summer to be compared. Out of the 150 animals, 13 (8.7%) were E. coli O157 shedders. The 13 ovine sorbitol-negative E. coli O157 were characterized by different PCR sets. The eae gene was detected in 11 isolate (85%), stx1 in 3 isolates (23%), stx2 in 8 isolates (62%), and finally the hlyA in 11 isolate (85%). Among the 13 isolates, 2 strains (15%) were positive for eae, stx1, stx2, and hlyA as gene combination, one isolate (8%) for eae, stx1, and hlyA, 5 isolates (38%) for eae, stx2, and hlyA, 1 isolate (8%) for eae and stx2, 2 isolates (15%) contained eae and hlyA, 1 isolate (8%) contained hlyA only, and finally, 1 isolate (8%) did not contain any of these genes. None of the isolates showed the gene combination eae stx1, stx1 hlyA, or stx2 hlyA. The results indicated significant association of unfavorable weather and management conditions on O157:H7 shedding while the age or sex did not play any role in this process.

Capturing Cognitive Fingerprints for Active Authentication

DTIC Science & Technology

2014-10-01

CAPTURING COGNITIVE FINGERPRINTS FOR ACTIVE AUTHENTICATION IOWA STATE UNIVERSITY OF SCIENCE & TECHNOLOGY OCTOBER 2014 FINAL TECHNICAL REPORT...REPORT TYPE FINAL TECHNICAL REPORT 3. DATES COVERED (From - To) SEP 2013 – APR 2014 4. TITLE AND SUBTITLE CAPTURING COGNITIVE FINGERPRINTS FOR ACTIVE...The project ended before the IRB application was approved. 15. SUBJECT TERMS Active Authentication, Cognitive Fingerprints , Biometric Modalities

Application of Communications Satellites to Educational Development. Final Technical Report, September 1, 1969-August 31, 1975.

ERIC Educational Resources Information Center

Morgan, Robert P.

Research is summarized in a brief final report built around a four-section bibliography. The first section lists periodic progress reports and articles which provide an overview of the program, including articles which pertain primarily to educational rather than technical aspects of satellite utilization. Theses carried out in the fields of…

Head Start Impact Study. Technical Report

ERIC Educational Resources Information Center

Puma, Michael; Bell, Stephen; Cook, Ronna; Heid, Camilla; Shapiro, Gary; Broene, Pam; Jenkins, Frank; Fletcher, Philip; Quinn, Liz; Friedman, Janet; Ciarico, Janet; Rohacek, Monica; Adams, Gina; Spier, Elizabeth

2010-01-01

This Technical Report is designed to provide technical detail to support the analysis and findings presented in the "Head Start Impact Study Final Report" (U.S. Department of Health and Human Services, January 2010). Chapter 1 provides an overview of the Head Start Impact Study and its findings. Chapter 2 provides technical information on the…

Matching Community and Technical College Professional/Technical Education Capacity to Employer Demand. Final Report.

ERIC Educational Resources Information Center

Sommers, Paul; Heg, Deena

A project was conducted to improve the state of Washington's community and technical college system by developing and using an improved occupational forecasting system to assess and respond to education and training needs. First, long-term occupational forecast data from Washington's Employment Security Department were matched with technical and…

75 FR 5784 - Guidance on Preparation of Market-Based Rate Filings and Electric Quarterly Reports by Public...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-02-04

... Preparation of Market-Based Rate Filings and Electric Quarterly Reports by Public Utilities; Notice of Technical Conference January 28, 2010. Take notice that Commission staff will convene a technical conference... final agenda of the technical conference. The March 3, 2010 technical conference will focus on the...

A Revision of Technical Mathematics Based on the NCTM Standards. Final Report.

ERIC Educational Resources Information Center

Near, Barbara

Between 1993 and 1996, Henry Ford Community College (Michigan) worked with business, industry, and technical instructors to revise their Technical Mathematics program in accordance with the National Council of Teachers of Mathematics (NCTM) Standards. The purpose of the project was to restructure the technical math curriculum and create a context…

76 FR 11327 - Technical Amendments to Rule 17a-8: Financial Recordkeeping and Reporting of Currency and Foreign...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-03-02

... SECURITIES AND EXCHANGE COMMISSION 17 CFR Part 240 [Release No. 34-63949] Technical Amendments to...: Securities and Exchange Commission. ACTION: Final rule; technical amendments. SUMMARY: The Securities and Exchange Commission (``Commission'') is adopting technical amendments to Rule 17a-8 under the Securities...

Final priority; technical assistance to improve state data capacity--National Technical Assistance Center to improve state capacity to accurately collect and report IDEA data. Final priority.

PubMed

2013-05-20

The Assistant Secretary for Special Education and Rehabilitative Services announces a priority under the Technical Assistance to Improve State Data Capacity program. The Assistant Secretary may use this priority for competitions in fiscal year (FY) 2013 and later years. We take this action to focus attention on an identified national need to provide technical assistance (TA) to States to improve their capacity to meet the data collection and reporting requirements of the Individuals with Disabilities Education Act (IDEA). We intend this priority to establish a TA center to improve State capacity to accurately collect and report IDEA data (Data Center).

Analysis for residual host cell proteins and DNA in process streams of a recombinant protein product expressed in Escherichia coli cells.

PubMed

Rathore, Anurag Singh; Sobacke, S E; Kocot, T J; Morgan, D R; Dufield, R L; Mozier, N M

2003-08-21

Analyses of crude samples from biotechnology processes are often required in order to demonstrate that residual host cell impurities are reduced or eliminated during purification. In later stages of development, as the processes are further developed and finalized, there is a tremendous volume of testing required to confirm the absence of residual host cell proteins (HCP) and DNA. Analytical tests for these components are very challenging since (1). they may be present at levels that span a million-fold range, requiring substantial dilutions; (2). are not a single component, often existing as fragments and a variety of structures; (3). require high sensitivity for final steps in process; and (4). are present in very complex matrices including other impurities, the product, buffers, salts and solvents. Due to the complex matrices and the variety of potential analytes, the methods of analysis are not truly quantitative for all species. Although these limitations are well known, the assays are still very much in demand since they are required for approval of new products. Methods for final products, described elsewhere, focus on approaches to achieve regulatory requirements. The study described herein will describe the technical rationale for measuring the clearance of HCP and DNA in the entire bioprocessing to purification from an Escherichia coli-derived expression system. Three analytical assays, namely, reversed-phase high-performance liquid chromatography (RP-HPLC), enzyme-linked immunosorbent assay (ELISA), and Threshold Total DNA Assay, were utilized to quantify the protein product, HCP and DNA, respectively. Product quantification is often required for yield estimation and is useful since DNA and HCP results are best expressed as a ratio to product for calculation of relative purification factors. The recombinant E. coli were grown to express the protein of interest as insoluble inclusion bodies (IB) within the cells. The IB were isolated by repeated homogenization and centrifugation and the inclusion body slurry (IBS) was solubilized with urea. After refolding the product, the solution was loaded on several commonly used ion exchangers (CM, SP, DEAE, and Q). Product was eluted in a salt gradient mode and fractions were collected and analyzed for product, HCP and DNA. The IBS used for this study contained about 15 mg/ml product, 38 mg/ml HCP and 1.1 mg/ml DNA. Thus, the relative amounts of HCP and DNA in the IBS was excessive, and about 10(3) times greater than typical (because the cells and IB were not processed with the normal number of washing steps during isolation). This was of interest since similar samples may be encountered when working with non-inclusion body systems, such as periplasmic expressions, or in cases where the upstream unit operations under-perform in IB cleaning. The study described herein describes the development of three robust methods that provide the essential process data needed. These findings are of general interest to other projects since applications of similar analytical technology may be used as a tool to develop processes, evaluate clearance of impurities, and produce a suitable product.

A comparison of Shiga-toxin 2 bacteriophage from classical enterohemorrhagic Escherichia coli serotypes and the German E. coli O104:H4 outbreak strain.

PubMed

Laing, Chad R; Zhang, Yongxiang; Gilmour, Matthew W; Allen, Vanessa; Johnson, Roger; Thomas, James E; Gannon, Victor P J

2012-01-01

Escherichia coli O104:H4 was associated with a severe foodborne disease outbreak originating in Germany in May 2011. More than 4000 illnesses and 50 deaths were reported. The outbreak strain was a typical enteroaggregative E. coli (EAEC) that acquired an antibiotic resistance plasmid and a Shiga-toxin 2 (Stx2)-encoding bacteriophage. Based on whole-genome phylogenies, the O104:H4 strain was most closely related to other EAEC strains; however, Stx2-bacteriophage are mobile, and do not necessarily share an evolutionary history with their bacterial host. In this study, we analyzed Stx2-bacteriophage from the E. coli O104:H4 outbreak isolates and compared them to all available Stx2-bacteriophage sequences. We also compared Stx2 production by an E. coli O104:H4 outbreak-associated isolate (ON-2011) to that of E. coli O157:H7 strains EDL933 and Sakai. Among the E. coli Stx2-phage sequences studied, that from O111:H- strain JB1-95 was most closely related phylogenetically to the Stx2-phage from the O104:H4 outbreak isolates. The phylogeny of most other Stx2-phage was largely concordant with their bacterial host genomes. Finally, O104:H4 strain ON-2011 produced less Stx2 than E. coli O157:H7 strains EDL933 and Sakai in culture; however, when mitomycin C was added, ON-2011 produced significantly more toxin than the E. coli O157:H7 strains. The Stx2-phage from the E. coli O104:H4 outbreak strain and the Stx2-phage from O111:H- strain JB1-95 likely share a common ancestor. Incongruence between the phylogenies of the Stx2-phage and their host genomes suggest the recent Stx2-phage acquisition by E. coli O104:H4. The increase in Stx2-production by ON-2011 following mitomycin C treatment may or may not be related to the high rates of hemolytic uremic syndrome associated with the German outbreak strain. Further studies are required to determine whether the elevated Stx2-production levels are due to bacteriophage or E. coli O104:H4 host related factors.

A Comparison of Shiga-Toxin 2 Bacteriophage from Classical Enterohemorrhagic Escherichia coli Serotypes and the German E. coli O104:H4 Outbreak Strain

PubMed Central

Laing, Chad R.; Zhang, Yongxiang; Gilmour, Matthew W.; Allen, Vanessa; Johnson, Roger; Thomas, James E.; Gannon, Victor P. J.

2012-01-01

Escherichia coli O104:H4 was associated with a severe foodborne disease outbreak originating in Germany in May 2011. More than 4000 illnesses and 50 deaths were reported. The outbreak strain was a typical enteroaggregative E. coli (EAEC) that acquired an antibiotic resistance plasmid and a Shiga-toxin 2 (Stx2)-encoding bacteriophage. Based on whole-genome phylogenies, the O104:H4 strain was most closely related to other EAEC strains; however, Stx2-bacteriophage are mobile, and do not necessarily share an evolutionary history with their bacterial host. In this study, we analyzed Stx2-bacteriophage from the E. coli O104:H4 outbreak isolates and compared them to all available Stx2-bacteriophage sequences. We also compared Stx2 production by an E. coli O104:H4 outbreak-associated isolate (ON-2011) to that of E. coli O157:H7 strains EDL933 and Sakai. Among the E. coli Stx2-phage sequences studied, that from O111:H- strain JB1-95 was most closely related phylogenetically to the Stx2-phage from the O104:H4 outbreak isolates. The phylogeny of most other Stx2-phage was largely concordant with their bacterial host genomes. Finally, O104:H4 strain ON-2011 produced less Stx2 than E. coli O157:H7 strains EDL933 and Sakai in culture; however, when mitomycin C was added, ON-2011 produced significantly more toxin than the E. coli O157:H7 strains. The Stx2-phage from the E. coli O104:H4 outbreak strain and the Stx2-phage from O111:H- strain JB1-95 likely share a common ancestor. Incongruence between the phylogenies of the Stx2-phage and their host genomes suggest the recent Stx2-phage acquisition by E. coli O104:H4. The increase in Stx2-production by ON-2011 following mitomycin C treatment may or may not be related to the high rates of hemolytic uremic syndrome associated with the German outbreak strain. Further studies are required to determine whether the elevated Stx2-production levels are due to bacteriophage or E. coli O104:H4 host related factors. PMID:22649523

Technical Evaluation Motor No. 10 (TEM-10)

NASA Technical Reports Server (NTRS)

1993-01-01

Technical Evaluation Motor No. 10 (TEM-10) was static fired on 27 Apr. 1993 at the Thiokol Corporation full-scale motor static test bay, T-24. This final test report documents the procedures, performance, and results of the static test firing of TEM-10. All observations, discussions, conclusions, and recommendations contained are final. Included is a presentation and discussion of TEM-10 performance, anomalies, and test results in concurrence with the objectives outlined in CTP-0110, Revision D, Space Shuttle Technical Evaluation Motor No. 10 (TEM-10) Static Fire Test Plan.

Final priority; Technical Assistance on State Data Collection--IDEA Data Management Center. Final priority.

PubMed

2014-08-05

The Assistant Secretary for the Office of Special Education and Rehabilitative Services (OSERS) announces a priority under the Technical Assistance on State Data Collection program. The Assistant Secretary may use this priority for competitions in fiscal year (FY) 2014 and later years. We take this action to fund a cooperative agreement to establish and operate an IDEA Data Management Center (Center) that will provide technical assistance (TA) to improve the capacity of States to meet the data collection requirements of the Individuals with Disabilities Education Act (IDEA).

Technical Topic 3.2.2.d Bayesian and Non-Parametric Statistics: Integration of Neural Networks with Bayesian Networks for Data Fusion and Predictive Modeling

DTIC Science & Technology

2016-05-31

and included explosives such as TATP, HMTD, RDX, RDX, ammonium nitrate , potassium perchlorate, potassium nitrate , sugar, and TNT. The approach...Distribution Unlimited UU UU UU UU 31-05-2016 15-Apr-2014 14-Jan-2015 Final Report: Technical Topic 3.2.2. d Bayesian and Non- parametric Statistics...of Papers published in non peer-reviewed journals: Final Report: Technical Topic 3.2.2. d Bayesian and Non-parametric Statistics: Integration of Neural

Porous media augmented with biochar for the retention of E. coli

NASA Astrophysics Data System (ADS)

Kolotouros, Christos A.; Manariotis, Ioannis D.; Karapanagioti, Hrissi K.

2016-04-01

A significant number of epidemic outbreaks has been attributed to waterborne fecal-borne pathogenic microorganisms from contaminated ground water. The transport of pathogenic microorganisms in groundwater is controlled by physical and chemical soil properties like soil structure, texture, percent water saturation, soil ionic strength, pore-size distribution, soil and solution pH, soil surface charge, and concentration of organic carbon in solution. Biochar can increase soil productivity by improving both chemical and physical soil properties. The mixing of biochar into soils may stimulate microbial population and activate dormant soil microorganisms. Furthermore, the application of biochar into soil affects the mobility of microorganisms by altering the physical and chemical properties of the soil, and by retaining the microorganisms on the biochar surface. The aim of this study was to investigate the effect of biochar mixing into soil on the transport of Escherichia coli in saturated porous media. Initially, batch experiments were conducted at two different ionic strengths (1 and 150 mM KCl) and at varying E. coli concentrations in order to evaluate the retention of E. coli on biochar in aqueous solutions. Kinetic analysis was conducted, and three isotherm models were employed to analyze the experimental data. Column experiments were also conducted in saturated sand columns augmented with different biochar contents, in order to examine the effect of biochar on the retention of E. coli. The Langmuir model fitted better the retention experimental data, compared to Freundlich and Tempkin models. The retention of E. coli was enhanced at lower ionic strength. Finally, biochar-augmented sand columns were more capable in retaining E. coli than pure sand columns.

Pathogenic Escherichia coli Found in Sewage Treatment Plants and Environmental Waters

PubMed Central

Anastasi, E. M.; Matthews, B.; Stratton, H. M.

2012-01-01

We previously demonstrated that some Escherichia coli strains with uropathogenic properties survived treatment stages of sewage treatment plants (STPs), suggesting that they may be released into the environment. We investigated the presence of such strains in the surrounding environmental waters of four STPs from which these persistent strains were isolated. In all, 264 E. coli isolates were collected from 129 receiving water sites in a 20-km radius surrounding STPs. We also included 93 E. coli strains collected from 18 animal species for comparison. Isolates were typed using a high-resolution biochemical fingerprinting method (the PhPlate system), and grouped into common (C) types. One hundred forty-seven (56%) environmental isolates were identical to strains found in STPs' final effluents. Of these, 140 (95%) carried virulence genes (VGs) associated with intestinal pathogenic E. coli (IPEC) or uropathogenic E. coli (UPEC) and were found in a variety of sites within areas sampled. Of the remaining 117 environmental strains not identical to STP strains, 105 belonged to 18 C types and 102 of them carried VGs found among IPEC or UPEC strains. These strains belonged mainly to phylogenetic groups A (A0 and A1) and B1 and to a lesser extent B22, B23, D1, and D2. Eight of 18 environmental C types, comprising 50 isolates, were also identical to bird strains. The presence of a high percentage of environmental E. coli in waters near STPs carrying VGs associated with IPEC and UPEC suggests that they may have derived from STP effluents and other nonpoint sources. PMID:22660714

Genetic Investigation of Beta-Lactam Associated Antibiotic Resistance Among Escherichia Coli Strains Isolated from Water Sources.

PubMed

Ranjbar, Reza; Sami, Mehrdad

2017-01-01

Antimicrobial resistance is an important factor threatening human health. It is widely accepted that antibiotic resistant bacteria such as Escherichia coli ( E. coli) released from humans and animals into the water sources, can introduce their resistance genes into the natural bacterial community. The aim of this study was to investigate the prevalence of bla TEM , bla CTX , bla SHV , bla OXA and bla VEB associated-antibiotic resistance among E. coli bacteria isolated from different water resources in Iran. The study contained all E. coli strains segregated from different surface water sources. The Kirby-Bauer method and combined discs method was determined in this study for testing antimicrobial susceptibility and strains that produced Extended-Spectrum Beta Lactamases (ESBL), respectively. DNA extraction kit was applied for genomic and plasmid DNA derivation. Finally the frequency of resistant genes including bla TEM , bla CTX , bla SHV , bla OXA and bla VEB in ESBL producing isolates were studied by PCR. One hundred E. coli strains were isolated and entered in the study. The highest antibiotic resistance was observed on clindamycin (96%). Moreover, 38.5% isolates were ESBL producers. The frequency of different ESBLs genes were 37%, 27%, 27%, and 25% for bla TEM , bla CTX , bla SHV , and bla OXA , respectively. The bla VEB wasn't found in any isolates. The study revealed a high prevalence of CTX-M, TEM, SHV and OXA genes among E. coli strains in surface water resources. In conclusion, these results raised a concern regarding the presence and distribution of these threatening factors in surface water sources and its subsequent outcomes.

The evolution of metabolic networks of E. coli

PubMed Central

2011-01-01

Background Despite the availability of numerous complete genome sequences from E. coli strains, published genome-scale metabolic models exist only for two commensal E. coli strains. These models have proven useful for many applications, such as engineering strains for desired product formation, and we sought to explore how constructing and evaluating additional metabolic models for E. coli strains could enhance these efforts. Results We used the genomic information from 16 E. coli strains to generate an E. coli pangenome metabolic network by evaluating their collective 76,990 ORFs. Each of these ORFs was assigned to one of 17,647 ortholog groups including ORFs associated with reactions in the most recent metabolic model for E. coli K-12. For orthologous groups that contain an ORF already represented in the MG1655 model, the gene to protein to reaction associations represented in this model could then be easily propagated to other E. coli strain models. All remaining orthologous groups were evaluated to see if new metabolic reactions could be added to generate a pangenome-scale metabolic model (iEco1712_pan). The pangenome model included reactions from a metabolic model update for E. coli K-12 MG1655 (iEco1339_MG1655) and enabled development of five additional strain-specific genome-scale metabolic models. These additional models include a second K-12 strain (iEco1335_W3110) and four pathogenic strains (two enterohemorrhagic E. coli O157:H7 and two uropathogens). When compared to the E. coli K-12 models, the metabolic models for the enterohemorrhagic (iEco1344_EDL933 and iEco1345_Sakai) and uropathogenic strains (iEco1288_CFT073 and iEco1301_UTI89) contained numerous lineage-specific gene and reaction differences. All six E. coli models were evaluated by comparing model predictions to carbon source utilization measurements under aerobic and anaerobic conditions, and to batch growth profiles in minimal media with 0.2% (w/v) glucose. An ancestral genome-scale metabolic model based on conserved ortholog groups in all 16 E. coli genomes was also constructed, reflecting the conserved ancestral core of E. coli metabolism (iEco1053_core). Comparative analysis of all six strain-specific E. coli models revealed that some of the pathogenic E. coli strains possess reactions in their metabolic networks enabling higher biomass yields on glucose. Finally the lineage-specific metabolic traits were compared to the ancestral core model predictions to derive new insight into the evolution of metabolism within this species. Conclusion Our findings demonstrate that a pangenome-scale metabolic model can be used to rapidly construct additional E. coli strain-specific models, and that quantitative models of different strains of E. coli can accurately predict strain-specific phenotypes. Such pangenome and strain-specific models can be further used to engineer metabolic phenotypes of interest, such as designing new industrial E. coli strains. PMID:22044664

IraL Is an RssB Anti-adaptor That Stabilizes RpoS during Logarithmic Phase Growth in Escherichia coli and Shigella

PubMed Central

Hryckowian, Andrew J.; Battesti, Aurelia; Lemke, Justin J.; Meyer, Zachary C.

2014-01-01

ABSTRACT RpoS (σS), the general stress response sigma factor, directs the expression of genes under a variety of stressful conditions. Control of the cellular σS concentration is critical for appropriately scaled σS-dependent gene expression. One way to maintain appropriate levels of σS is to regulate its stability. Indeed, σS degradation is catalyzed by the ClpXP protease and the recognition of σS by ClpXP depends on the adaptor protein RssB. Three anti-adaptors (IraD, IraM, and IraP) exist in Escherichia coli K-12; each interacts with RssB and inhibits RssB activity under different stress conditions, thereby stabilizing σS. Unlike K-12, some E. coli isolates, including uropathogenic E. coli strain CFT073, show comparable cellular levels of σS during the logarithmic and stationary growth phases, suggesting that there are differences in the regulation of σS levels among E. coli strains. Here, we describe IraL, an RssB anti-adaptor that stabilizes σS during logarithmic phase growth in CFT073 and other E. coli and Shigella strains. By immunoblot analyses, we show that IraL affects the levels and stability of σS during logarithmic phase growth. By computational and PCR-based analyses, we reveal that iraL is found in many E. coli pathotypes but not in laboratory-adapted strains. Finally, by bacterial two-hybrid and copurification analyses, we demonstrate that IraL interacts with RssB by a mechanism distinct from that used by other characterized anti-adaptors. We introduce a fourth RssB anti-adaptor found in E. coli species and suggest that differences in the regulation of σS levels may contribute to host and niche specificity in pathogenic and nonpathogenic E. coli strains. PMID:24865554

Non-standard amino acid incorporation into proteins using Escherichia coli cell-free protein synthesis

NASA Astrophysics Data System (ADS)

Hong, Seok Hoon; Kwon, Yong-Chan; Jewett, Michael

2014-06-01

Incorporating non-standard amino acids (NSAAs) into proteins enables new chemical properties, new structures, and new functions. In recent years, improvements in cell-free protein synthesis (CFPS) systems have opened the way to accurate and efficient incorporation of NSAAs into proteins. The driving force behind this development has been three-fold. First, a technical renaissance has enabled high-yielding (>1 g/L) and long-lasting (>10 h in batch operation) CFPS in systems derived from Escherichia coli. Second, the efficiency of orthogonal translation systems has improved. Third, the open nature of the CFPS platform has brought about an unprecedented level of control and freedom of design. Here, we review recent developments in CFPS platforms designed to precisely incorporate NSAAs. In the coming years, we anticipate that CFPS systems will impact efforts to elucidate structure/function relationships of proteins and to make biomaterials and sequence-defined biopolymers for medical and industrial applications.

A second-generation expression system for tyrosine-sulfated proteins and its application in crop protection.

PubMed

Schwessinger, Benjamin; Li, Xiang; Ellinghaus, Thomas L; Chan, Leanne Jade G; Wei, Tong; Joe, Anna; Thomas, Nicholas; Pruitt, Rory; Adams, Paul D; Chern, Maw Sheng; Petzold, Christopher J; Liu, Chang C; Ronald, Pamela C

2016-04-18

Posttranslational modification (PTM) of proteins and peptides is important for diverse biological processes in plants and animals. The paucity of heterologous expression systems for PTMs and the technical challenges associated with chemical synthesis of these modified proteins has limited detailed molecular characterization and therapeutic applications. Here we describe an optimized system for expression of tyrosine-sulfated proteins in Escherichia coli and its application in a bio-based crop protection strategy in rice.

A second-generation expression system for tyrosine-sulfated proteins and its application in crop protection

DOE PAGES

Schwessinger, Benjamin; Li, Xiang; Ellinghaus, Thomas L.; ...

2015-11-27

Posttranslational modification (PTM) of proteins and peptides is important for diverse biological processes in plants and animals. The paucity of heterologous expression systems for PTMs and the technical challenges associated with chemical synthesis of these modified proteins has limited detailed molecular characterization and therapeutic applications. Here we describe an optimized system for expression of tyrosine-sulfated proteins in Escherichia coli and its application in a bio-based crop protection strategy in rice.

Final June Revisions Rule Significant Contribution Assessment TSD

EPA Pesticide Factsheets

This Technical Support Document (TSD) presents quantitative assessments of the relationship between the final February revisions to the Transport Rule, the final June revisions rule, and the original analysis conducted for the final Transport Rule.

Develop a Comprehensive Technical Training and Data Collection Program for Structural Welders and Fitters: Technical Proposal

DTIC Science & Technology

2006-05-30

implementation Final Report 4 TECHNICAL PLAN AND RESULTS Task 1: Initiate the Project Management System Two senior NGSS production management...1 Technical Plan and Results...Third the system is hosted on a handheld unit which provides the foremen with an efficient daily planning tool. The Pilot System which entails

Laser Technician Associate Degree Program. A Proposal Submitted to Wisconsin State Board of Vocational, Technical, and Adult Education. (Curriculum Development.) Final Report.

ERIC Educational Resources Information Center

North Central Technical Inst., Wausau, WI.

This final report contains the program proposal with supporting data for developing curriculum materials for and implementing an associate-degree laser technology program at the North Central Technical Institute. The proposal outline provides this information: (1) objectives for the program designed to prepare a technician to safely operate,…

California Deaf-Blind Services Final Report, October 1, 1995-September 30, 1999 [and] October 1, 1999-June 30, 2000 (No Cost Extension).

ERIC Educational Resources Information Center

Goertz, Lori; Franklin, Barbara

This final report describes the activities and outcomes of the California Deaf-Blind Services (CDBS) program, a regionally based, family focused technical assistance and training project designed to improve services to children with deaf-blindness. The project conducted the following activities: (1) provided technical assistance to families and…

New Jersey Technical Assistance Project (N.J. TAP): "Educational Resources and Support Services on Behalf of Children with Multiple Sensory Impairment." Final Report.

ERIC Educational Resources Information Center

New Jersey State Dept. of Education, Trenton.

This final report describes activities and accomplishments of the New Jersey Technical Assistance Project, a project to improve educational resources and support services for students with multiple sensory impairment (deaf-blindness). Activities and accomplishments are presented in a tabular format for each project goal and objective. The project…

78 FR 77563 - Technical Amendments

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-24

... NATIONAL CREDIT UNION ADMINISTRATION 12 CFR Parts 700, 701, and 704 RIN 3133-AE33 Technical Amendments AGENCY: National Credit Union Administration (NCUA). ACTION: Final rule. SUMMARY: The NCUA Board... credit unions. The technical amendments conform the regulations to a recent policy change adopted by the...

75 FR 33682 - Export Administration Regulations; Technical Amendments

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-15

...-01] RIN 0694-AE93 Export Administration Regulations; Technical Amendments AGENCY: Bureau of Industry... Bureau of Industry and Security (BIS) makes a technical amendment to the Export Administration... review of final decisions and orders issued in BIS export control administrative enforcement proceedings...

78 FR 41331 - Defense Federal Acquisition Regulation Supplement; Technical Amendments

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-10

... DEPARTMENT OF DEFENSE Defense Acquisition Regulations System 48 CFR Part 225 Defense Federal Acquisition Regulation Supplement; Technical Amendments AGENCY: Defense Acquisition Regulations System, Department of Defense (DoD). ACTION: Final rule. SUMMARY: DoD is making technical amendment to the Defense...

Construction of a Specialized Cloning Strain of E. Coli for the Nitrate Reductase Genes of Haloferax Denitrificans

NASA Technical Reports Server (NTRS)

Johnson, Emmett

1999-01-01

This is the final report on Joint Research Interchange (NCC2-5011) "Construction of a Specialized Cloning Strain of E.. coli for the Nitrate Reductase Genes of Haloferax denitrificans." Originally the award was 11/l/93-10/31/95, but there were no-cost extensions made, because of a year Sabbatical at the Pasteur Institute in Paris and other leaves of 3 months each at the Pasteur Institute, during which work could not be done on this project, which extended the closing date to 10/30/98.

Structural basis for suppression of hypernegative DNA supercoiling by E. coli topoisomerase I

DOE PAGES

Tan, Kemin; Zhou, Qingxuan; Cheng, Bokun; ...

2015-10-20

Escherichia coli topoisomerase I has an essential function in preventing hypernegative supercoiling of DNA. A full length structure of E. coli topoisomerase I reported here shows how the C-terminal domains bind single-stranded DNA (ssDNA) to recognize the accumulation of negative supercoils in duplex DNA. These C-terminal domains of E. coli topoisomerase I are known to interact with RNA polymerase, and two flexible linkers within the C-terminal domains may assist in the movement of the ssDNA for the rapid removal of transcription driven negative supercoils. The structure has also unveiled for the first time how the 4-Cys zinc ribbon domain andmore » zinc ribbon-like domain bind ssDNA with primarily π -stacking interactions. Finally, this novel structure, in combination with new biochemical data, provides important insights into the mechanism of genome regulation by type IA topoisomerases that is essential for life, as well as the structures of homologous type IA TOP3α and TOP3β from higher eukaryotes that also have multiple 4-Cys zinc ribbon domains required for their physiological functions.« less

The presence of Enterococcus, coliforms and E. coli in a commercial yeast manufacturing process.

PubMed

O'Brien, S S; Lindsay, D; von Holy, A

2004-07-01

This study evaluated a typical commercial yeast manufacturing process for bacterial contamination. Product line samples of a commercial yeast manufacturing process and the corresponding seed yeast manufacturing process were obtained upstream from the final compressed and dry yeast products. All samples were analysed before (non-PI) and after preliminary incubation (PI) at 37 degrees C for 24 h. The PI procedure was incorporated for amplification of bacterial counts below the lower detection limit. Enterococcus, coliform and Escherichia coli counts were quantified by standard pour-plate techniques using selective media. Presence at all stages and progressive increases in counts of Enterococcus, coliforms and E. coli during processing in the commercial manufacturing operation suggested that the primary source of contamination of both compressed and dry yeast with these bacteria was the seed yeast manufacturing process and that contamination was amplified throughout the commercial yeast manufacturing process. This was confirmed by surveys of the seed yeast manufacturing process which indicated that contamination of the seed yeast with Enterococcus, coliforms and E. coli occurred during scale up of seed yeast biomass destined as inoculum for the commercial fermentation.

Antibiotic Resistance of Bacteria Involved in Urinary Infections in Brazil: A Cross-Sectional and Retrospective Study

PubMed Central

Rodrigues, Wellington Francisco; Miguel, Camila Botelho; Nogueira, Ana Paula Oliveira; Ueira-Vieira, Carlos; Paulino, Tony De Paiva; Soares, Siomar De Castro; De Resende, Elisabete Aparecida Mantovani Rodrigues; Lazo-Chica, Javier Emilio; Araújo, Marcelo Costa; Oliveira, Carlo José

2016-01-01

Empirical and prolonged antimicrobial treatment of urinary tract infections caused by Escherichia coli is associated with the emergence of bacterial resistance, and not all countries have strict policies against the indiscriminate use of drugs in order to prevent resistance. This cross-sectional and retrospective study (2010–2015) aimed to evaluate the sensitivity and resistance of patient-derived E. coli to different drugs broadly used to treat urinary infections in Brazil: ampicillin + sulbactam, cephalothin, ciprofloxacin, norfloxacin, and nitrofurantoin. We obtained 1654 E. coli samples from ambulatory patients with disease symptoms of the urinary tract from a Brazilian public hospital. While all antibiotics were effective in killing E. coli to a large degree, nitrofurantoin was the most effective, with fewer samples exhibiting antibiotic resistance. We assessed the costs of generic and brand name versions of each antibiotic. Nitrofurantoin, the most effective antibiotic, was the cheapest, followed by the fluoroquinolones (ciprofloxacin and norfloxacin), ampicillin + sulbactam and, lastly, cephalothin. Finally, assessment of antibiotic resistance to fluoroquinolones over the study period and extrapolation of the data led to the conclusion that these antibiotics could no longer be effective against E. coli-based urinary infections in approximately 20 years if their indiscriminate use in empirical treatment continues. PMID:27649224

A mixture of Lactobacillus casei, Lactobacillus lactis, and Paenibacillus polymyxa reduces Escherichia coli O157:H7 in finishing feedlot cattle.

PubMed

Stanford, Kim; Bach, Susan; Baah, John; McAllister, Tim

2014-05-01

A direct-fed microbial (DFM) containing Paenibacillus polymyxa, Lactobacillus casei, and Lactobacillus lactis was fed to cattle (n = 120) to determine impacts on shedding and survival of Escherichia coli O157:H7 in feces. Cattle were individually penned and fed diets containing 0 (control), 4 × 10(7) CFU (DFM-4), 8 × 10(7) CFU (DFM-8), or 1.2 × 10(8) CFU (DFM-12) lactobacilli per kg of dietary dry matter over 84-day fall-winter growing and 140-day spring-summer finishing periods. Fecal grab samples were collected from cattle at 28-day intervals, E. coli O157:H7 was detected by immunomagnetic separation, and isolates were compared by pulsed-field gel electrophoresis. During the growing period, feces negative for E. coli O157 from each dietary treatment were inoculated with 10(5) CFU/g nalidixic acid-resistant E. coli O157:H7 and were incubated at 4 and 22(u) C for 11 weeks. Fecal pH and fecal dry matter were measured on days 0, 1, 3, and 7 and weekly thereafter, with E. coli O157:H7 enumerated through dilution plating. Treatment with DFMs did not affect survival of E. coli O157:H7 in feces or fecal pH (P > 0.05). Only one steer was positive for E. coli O157:H7 during the growing period, but during the finishing period, DFM-8 and DFM-12 reduced the prevalence of E. coli O157:H7 in feces (P < 0.05). Feeding DFMs also reduced the frequency of individual steers shedding E. coli O157:H7 during finishing (P < 0.05), with control steers shedding E. coli O157:H7 up to four times, whereas DFM-12 steers shed E. coli O157:H7 a maximum of twice. Treatment with DFMs influenced pulsed-field gel electrophoresis profiles; steers that were fed DFM-8 and DFM-12 shed more diverse subtypes of E. coli O157:H7 than did control or DFM-4 steers. Because a companion study found linear improvement in performance with increasing dosage of DFMs in the first 28 days of the growing period, targeted use of DFM-12 during this time and for the final 1 or 2 weeks prior to slaughter may optimize performance and reduce E. coli O157:H7 while minimizing feed costs.

Final Revisions Rule Significant Contribution Assessment TSD

EPA Pesticide Factsheets

This Technical Support Document (TSD) presents quantitative assessments of the relationship between final revisions to the Transport Rule and the original analysis conducted for the final Transport Rule.

Reduction of extended-spectrum-β-lactamase- and AmpC-β-lactamase-producing Escherichia coli through processing in two broiler chicken slaughterhouses.

PubMed

Pacholewicz, Ewa; Liakopoulos, Apostolos; Swart, Arno; Gortemaker, Betty; Dierikx, Cindy; Havelaar, Arie; Schmitt, Heike

2015-12-23

Whilst broilers are recognised as a reservoir of extended-spectrum-β-lactamase (ESBL)- and AmpC-β-lactamase (AmpC)-producing Escherichia coli, there is currently limited knowledge on the effect of slaughtering on its concentrations on poultry meat. The aim of this study was to establish the concentration of ESBL/AmpC producing E. coli on broiler chicken carcasses through processing. In addition the changes in ESBL/AmpC producing E. coli concentrations were compared with generic E. coli and Campylobacter. In two slaughterhouses, the surface of the whole carcasses was sampled after 5 processing steps: bleeding, scalding, defeathering, evisceration and chilling. In total, 17 batches were sampled in two different slaughterhouses during the summers of 2012 and 2013. ESBL/AmpC producing E. coli was enumerated on MacConkey agar with 1mg/l cefotaxime, and the ESBL/AmpC phenotypes and genotypes were characterised. The ESBL/AmpC producing E. coli concentrations varied significantly between the incoming batches in both slaughterhouses. The concentrations on broiler chicken carcasses were significantly reduced during processing. In Slaughterhouse 1, all subsequent processing steps reduced the concentrations except evisceration which led to a slight increase that was statistically not significant. The changes in concentration between processing steps were relatively similar for all sampled batches in this slaughterhouse. In contrast, changes varied between batches in Slaughterhouse 2, and the overall reduction through processing was higher in Slaughterhouse 2. Changes in ESBL/AmpC producing E. coli along the processing line were similar to changes in generic E. coli in both slaughterhouses. The effect of defeathering differed between ESBL/AmpC producing E. coli and Campylobacter. ESBL/AmpC producing E. coli decreased after defeathering, whereas Campylobacter concentrations increased. The genotypes of ESBL/AmpC producing E. coli (blaCTX-M-1, blaSHV-12, blaCMY-2, blaTEM-52c, blaTEM-52cvar) from both slaughterhouses match typical poultry genotypes. Their distribution differed between batches and changed throughout processing for some batches. The concentration levels found after chilling were between 10(2) and 10(5)CFU/carcass. To conclude, changes in ESBL/AmpC producing E. coli concentrations on broiler chicken carcasses during processing are influenced by batch and slaughterhouse, pointing to the role of both primary production and process control for reducing ESBL/AmpC producing E. coli levels in final products. Due to similar changes upon processing, E. coli can be used as a process indicator of ESBL/AmpC producing E. coli, because the processing steps had similar impact on both organisms. Cross contamination may potentially explain shifts in genotypes within some batches through the processing. Copyright © 2015 Elsevier B.V. All rights reserved.

Social Security cost-of-living adjustments and the Consumer Price Index.

PubMed

Burdick, Clark; Fisher, Lynn

2007-01-01

OASDI benefits are indexed for inflation to protect beneficiaries from the loss of purchasing power implied by inflation. In the absence of such indexing, the purchasing power of Social Security benefits would be eroded as rising prices raise the cost of living. By statute, cost-of-living adjustments (COLAs) for Social Security benefits are calculated using the Bureau of Labor Statistics (BLS) Consumer Price Index for Urban Wage Earners and Clerical Workers (CPI-W). Some argue that this index does not accurately reflect the inflation experienced by the elderly population and should be changed to an elderly-specific price index such as the Experimental Consumer Price Index for Americans 62 Years of Age and Older, often referred to as the Consumer Price Index for the Elderly (CPI-E). Others argue that the measure of inflation underlying the COLA is technically biased, causing it to overestimate changes in the cost of living. This argument implies that current COLAs tend to increase, rather than merely maintain, the purchasing power of benefits over time. Potential bias in the CPI as a cost-of-living index arises from a number of sources, including incomplete accounting for the ability of consumers to substitute goods or change purchasing outlets in response to relative price changes. The BLS has constructed a new index called the Chained Consumer Price Index for All Urban Consumers (C-CPI-U) that better accounts for those consumer adjustments. Price indexes are not true cost-of-living indexes, but approximations of cost-of-living indexes (COLI). The Bureau of Labor Statistics (2006a) explains the difference between the two: As it pertains to the CPI, the COLI for the current month is based on the answer to the following question: "What is the cost, at this month ' market prices, of achieving the standard of living actually attained in the base period?" This cost is a hypothetical expenditure-the lowest expenditure level necessary at this month's prices to achieve the base-period's living standard.... Unfortunately, because the cost of achieving a living standard cannot be observed directly, in operational terms, a COLI can only be approximated. Although the CPI cannot be said to equal a cost-of-living index, the concept of the COLI provides the CPI's measurement objective and the standard by which we define any bias in the CPI. While all versions of the CPI only approximate the actual changes in the cost of living, the CPI-E has several additional technical limitations. First, the CPI-E may better account for the goods and services typically purchased by the elderly, but the expenditure weights for the elderly are the only difference between the CPI-E and CPI-W. These weights are based on a much smaller sample than the other two indices, making it less precise. Second, the CPI-E does not account for differences in retail outlets frequented by the aged population or the prices they pay. Finally, the purchasing population measured in the CPI-E is not necessarily identical to the Social Security beneficiary population, where more than one-fifth of OASDI beneficiaries are under age 62. Likewise, over one-fifth of persons aged 62 or older are not beneficiaries, but they are included in the CPI-E population. Finally, changes in the index used to calculate COLAs directly affect the amount of benefits paid, and as a result, projected solvency of the Social Security program. A switch to the CPI-E for the December 2006 COLA (received in January 2007) would have resulted in an average monthly benefit $0.90 higher than that received. If the December 2006 COLA had been adjusted by the Chained CPI-U instead, the average monthly benefit would have been $4.70 less than with current indexing. Any changes to the COLA that would cause faster growth in individual benefits would make the projected date of insolvency sooner, while slower growth would delay insolvency. Hobijn and Lagakos (2003) estimated that switching to the CPI-E for COLAs would move projected insolvency sooner by 3-5 years. A projection by SSA's Office of the Chief Actuary estimated that annual COLAs based on the Chained C-CPI-U beginning in 2006 would delay the date of OASDI insolvency by 4 years.

75 FR 15342 - Advisory Committees; Technical Amendment

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-29

... DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration 21 CFR Part 14 [Docket No. FDA-2010-N-0001] Advisory Committees; Technical Amendment Agency: Food and Drug Administration, HHS. ACTION: Final rule; technical amendment. SUMMARY: The Food and Drug Administration (FDA) is amending its...

A GENOMICALLY MODIFIED MARKER STRAIN OF ESCHERICHIA COLI. (R825354)

EPA Science Inventory

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

CymA and Exogenous Flavins Improve Extracellular Electron Transfer and Couple It to Cell Growth in Mtr-Expressing Escherichia coli

DOE PAGES

Jensen, Heather M.; TerAvest, Michaela A.; Kokish, Mark G.; ...

2016-03-22

Introducing extracellular electron transfer pathways into heterologous organisms offers the opportunity to explore fundamental biogeochemical processes and to biologically alter redox states of exogenous metals for various applications. While expression of the MtrCAB electron nanoconduit from Shewanella oneidensis MR-1 permits extracellular electron transfer in Escherichia coli, the low electron flux and absence of growth in these cells limits their practicality for such applications. In this paper, we investigate how the rate of electron transfer to extracellular Fe(III) and cell survival in engineered E. coli are affected by mimicking different features of the S. oneidensis pathway: the number of electron nanoconduits,more » the link between the quinol pool and MtrA, and the presence of flavin-dependent electron transfer. While increasing the number of pathways does not significantly improve the extracellular electron transfer rate or cell survival, using the native inner membrane component, CymA, significantly improves the reduction rate of extracellular acceptors and increases cell viability. Strikingly, introducing both CymA and riboflavin to Mtr-expressing E. coli also allowed these cells to couple metal reduction to growth, which is the first time an increase in biomass of an engineered E. coli has been observed under Fe 2O 3 (s) reducing conditions. Overall and finally, this work provides engineered E. coli strains for modulating extracellular metal reduction and elucidates critical factors for engineering extracellular electron transfer in heterologous organisms.« less

CymA and Exogenous Flavins Improve Extracellular Electron Transfer and Couple It to Cell Growth in Mtr-Expressing Escherichia coli

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jensen, Heather M.; TerAvest, Michaela A.; Kokish, Mark G.

Introducing extracellular electron transfer pathways into heterologous organisms offers the opportunity to explore fundamental biogeochemical processes and to biologically alter redox states of exogenous metals for various applications. While expression of the MtrCAB electron nanoconduit from Shewanella oneidensis MR-1 permits extracellular electron transfer in Escherichia coli, the low electron flux and absence of growth in these cells limits their practicality for such applications. In this paper, we investigate how the rate of electron transfer to extracellular Fe(III) and cell survival in engineered E. coli are affected by mimicking different features of the S. oneidensis pathway: the number of electron nanoconduits,more » the link between the quinol pool and MtrA, and the presence of flavin-dependent electron transfer. While increasing the number of pathways does not significantly improve the extracellular electron transfer rate or cell survival, using the native inner membrane component, CymA, significantly improves the reduction rate of extracellular acceptors and increases cell viability. Strikingly, introducing both CymA and riboflavin to Mtr-expressing E. coli also allowed these cells to couple metal reduction to growth, which is the first time an increase in biomass of an engineered E. coli has been observed under Fe 2O 3 (s) reducing conditions. Overall and finally, this work provides engineered E. coli strains for modulating extracellular metal reduction and elucidates critical factors for engineering extracellular electron transfer in heterologous organisms.« less

2012 Technical Corrections Fact Sheet

EPA Pesticide Factsheets

Final Rule: 2012 Technical Corrections, Clarifying and Other Amendments to theGreenhouse Gas Reporting Rule, and Confidentiality Determinations for Certain DataElements of the Fluorinated Gas Source Category

Improving formaldehyde consumption drives methanol assimilation in engineered E. coli.

PubMed

Woolston, Benjamin M; King, Jason R; Reiter, Michael; Van Hove, Bob; Stephanopoulos, Gregory

2018-06-19

Due to volatile sugar prices, the food vs fuel debate, and recent increases in the supply of natural gas, methanol has emerged as a promising feedstock for the bio-based economy. However, attempts to engineer Escherichia coli to metabolize methanol have achieved limited success. Here, we provide a rigorous systematic analysis of several potential pathway bottlenecks. We show that regeneration of ribulose 5-phosphate in E. coli is insufficient to sustain methanol assimilation, and overcome this by activating the sedoheptulose bisphosphatase variant of the ribulose monophosphate pathway. By leveraging the kinetic isotope effect associated with deuterated methanol as a chemical probe, we further demonstrate that under these conditions overall pathway flux is kinetically limited by methanol dehydrogenase. Finally, we identify NADH as a potent kinetic inhibitor of this enzyme. These results provide direction for future engineering strategies to improve methanol utilization, and underscore the value of chemical biology methodologies in metabolic engineering.

Imipenem-resistance in Serratia marcescens is mediated by plasmid expression of KPC-2.

PubMed

Su, W-Q; Zhu, Y-Q; Deng, N-M; Li, L

2017-04-01

Imipenem is a broad-spectrum carbapenem antibiotic with applications against severe bacterial infections. Here, we describe the identification of imipenem-resistant Serratia marcescens in our hospital and the role of plasmid-mediated KPC-2 expression in imipenem resistance. We used the modified Hodge test to detect carbapenemase produced in imipenem-resistant strains. His resistance can be transferred to E. coli in co-culture tests, which implicates the plasmid in imipenem resistance. PCR amplification from the plasmid identified two products consistent with KPC-2 of 583 and 1050 bp that were also present in E. coli after co-culture. The restriction pattern for both plasmids was identical, supporting the transfer from the S. marcescens isolate to E. coli. Finally, gene sequencing confirmed KPC-2 in the plasmid. Due to the presence of KPC-2 in the imipenem-resistant S. marcescens, we propose that KPC-2 mediates antibiotic resistance in the S. marcescens isolate.

Effect of disinfection upon dissolved organic carbon (DOC) in wastewater: bacterial bioassays.

PubMed

Arana, I; Santorum, P; Muela, A; Barcina, I

2000-08-01

Quantitative and qualitative changes in organic matter content of wastewater effluents attributable to chlorination and ozonation have been analysed using bioassays as well as organic carbon direct measures. Bioassays were carried out using the bacterial populations of wastewater and two Escherichia coli strains as test micro-organisms. Our results indicate that pure strains present some advantages over indigenous bacteria. Although wastewater bacterial populations are better adapted to growth in wastewater, E. coli strains are more sensitive to changes in dissolved organic carbon (DOC) content. Moreover, the use of pure cultures allows estimation of the portion of DOC which can be converted in cell biomass, the assimilable organic carbon (AOC). Finally, the results obtained using prototrophic and the auxotrophic strains of E. coli suggested that ozonation alters the amino acid composition of wastewater while chlorination does not change the quantity nor the quality of the DOC present in effluents.

Evaluation of Routine Atmospheric Sounding Measurements using Unmanned Systems (ERASMUS): Final Technical Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

de Boer, Gijs; Lawrence, Dale; Palo, Scott

2017-03-29

This final technical report details activities undertaken as part of the referenced project. Included is information on the preparation of aircraft for deployment to Alaska, summaries of the three deployments covered under this project, and a brief description of the dataset and science directions pursued. Additionally, we provide information on lessons learned, publications, and presentations resulting from this work.

Vocational Curriculum Modification. Teaching Technical Language to Learning Handicapped Students. Project HIRE: A Curriculum Management System for Instructing the Handicapped. Final Report, Volume 1.

ERIC Educational Resources Information Center

Gardner, David C.; And Others

Volume 1 of the final report on Project HIRE reports the design, development, field-testing, and refining of self-instructional packages to teach entry level technical vocabulary to learning handicapped students mainstreamed in vocational programs. Volume 2, a management handbook, reports the methods and findings concerning development of…

Joint Technical Architecture for Robotic Systems (JTARS)-Final Report

NASA Technical Reports Server (NTRS)

Bradley, Arthur T.; Holloway, Sidney E., III

2006-01-01

This document represents the final report for the Joint Technical Architecture for Robotic Systems (JTARS) project, funded by the Office of Exploration as part of the Intramural Call for Proposals of 2005. The project was prematurely terminated, without review, as part of an agency-wide realignment towards the development of a Crew Exploration Vehicle (CEV) and meeting the near-term goals of lunar exploration.

Review of Final Year MSP Evaluations, Performance Period 2007. Analytic and Technical Support for Mathematics and Science Partnerships

ERIC Educational Resources Information Center

Bobronnikov, Ellen; Rhodes, Hilary; Bradley, Cay

2010-01-01

This final report culminates the evaluation and technical assistance provided for the U.S. Department of Education's Mathematics and Science Partnership (MSP) Program and its projects since 2005. As part of this support, Abt Associates looked across the portfolio of projects funded by the MSP program to draw lessons on best practices. This…

Pacific Educational Computer Network Study. Final Report.

ERIC Educational Resources Information Center

Hawaii Univ., Honolulu. ALOHA System.

The Pacific Educational Computer Network Feasibility Study examined technical and non-technical aspects of the formation of an international Pacific Area computer network for higher education. The technical study covered the assessment of the feasibility of a packet-switched satellite and radio ground distribution network for data transmission…

Studying the growth boundary and subsequent time to growth of pathogenic Escherichia coli serotypes by turbidity measurements.

PubMed

Valero, A; Rodríguez, M; Carrasco, E; Pérez-Rodríguez, F; García-Gimeno, R M; Zurera, G

2010-09-01

The presence of Escherichia coli in contaminated food products is commonly attributed to faecal contamination when they are improperly handled and/or when inactivation treatments fail. Adaptation of E. coli at low pH and a(w) levels can vary at different temperatures depending on the serotype, thus more detailed studies are needed. In this work, a screening to assess the growth of four pathogenic serotypes of E. coli (O55:H6; O59:H21; O158:H23 and O157:H7) was performed. Subsequently, boundary models were elaborated with the fastest serotype selected at different temperatures (8, 12 and 16 degrees C), and inoculum levels (2, 3 and 4log cfu/mL) as function of pH (7.00-5.00) and a(w) (0.999-0.960). Finally, the growth kinetics of E. coli was described in the conditions that allowed growth. Results obtained showed that the serotypes O157:H7 and O59:H21 did not grow at more stringent conditions (8 degrees C; pH 5.50), while the E. coli O158:H23 was the best adapted, resulting in faster growth. The logistic regression models presented a good adjustment to data observed since more than 96.7% of cases were correctly classified. The growth interface was shifted to more limited conditions as the inoculum size was higher. Detection times (t(d), h) and their variability were higher at low levels of the environmental factors studied. This work provides insight on the growth kinetics of E. coli at various environmental conditions. Copyright 2010 Elsevier Ltd. All rights reserved.

GENETIC ENGINEERING OF ESCHERICHIA COLI FOR ENHANCED BIOACCUMULATION OF MERCURY. (R827227)

EPA Science Inventory

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

ADHESION FORCES BETWEEN E. COLI BACTERIA AND BIOMATERIAL SURFACES (R821268)

EPA Science Inventory

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

Community College Technical Mathematics Project. Final Report.

ERIC Educational Resources Information Center

Self, Samuel L.

The purpose of the research project was to develop an applied or technical mathematics curriculum which would meet the needs of vocational-technical students at the community college level. The research project was divided into three distinct phases: Identifying the mathematical concepts requisite for job-entry competencies in each of the…

7 CFR 614.10 - Appeals before the Farm Service Agency county committee.

Code of Federal Regulations, 2010 CFR

2010-01-01

... section are completed, provide the FSA county committee with a written technical determination in the form... part 780, a participant may appeal a final technical determination or a program decision to the FSA... appeal requests review of the technical determination by the applicable State Conservationist prior to...

77 FR 47495 - Final Priority; Technical Assistance on State Data Collection, Analysis, and Reporting-National...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-08-08

... intervention and preschool service providers with data on their qualifications, certification, and preparation... Priority; Technical Assistance on State Data Collection, Analysis, and Reporting--National IDEA Technical Assistance Center on Early Childhood Longitudinal Data Systems; Rule #0;#0;Federal Register / Vol. 77, No...

77 FR 18716 - Transportation Security Administration Postal Zip Code Change; Technical Amendment

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-28

... organizational changes and it has no substantive effect on the public. DATES: Effective March 28, 2012. FOR... No. 1572-9] Transportation Security Administration Postal Zip Code Change; Technical Amendment AGENCY: Transportation Security Administration, DHS. ACTION: Final rule. SUMMARY: This rule is a technical change to...

Survival and distribution of Escherichia coli on diverse fresh-cut baby leafy greens under preharvest through postharvest conditions.

PubMed

Tomás-Callejas, Alejandro; López-Velasco, Gabriela; Camacho, Alex B; Artés, Francisco; Artés-Hernández, Francisco; Suslow, Trevor V

2011-12-02

Escherichia coli O157:H7 has been associated in multiple outbreaks linked to the consumption of whole produce and fresh-cut leafy vegetables. However, plant-based foods had not been traditionally recognized as a host for enteric pathogens until the elevated incidence of produce-related outbreaks became apparent. The survival dynamics of two cocktails of generic E. coli (environmental water, plant and soil isolates) and E. coli O157:H7 within the phyllosphere of Mizuna, Red Chard and Tatsoi during their production, harvest, minimal processing, packaging and storage over two greenhouse production cycles were studied. Genotyping of applied generic E. coli strains to evaluate their comparative survival and relative abundance in the phyllosphere by REP-PCR is also reported. The Mizuna, Red Chard and Tatsoi shoots were grown under standard greenhouse conditions and fertility management. Both E. coli cocktails were spray-inoculated separately and determined to result in an initial mean population density of log 4.2 CFU/cm². Leaves were harvested as mini-greens approximating commercial maturity, minimally processed in a model washing system treated with 3 mg/L of ClO₂ and stored for 7 days at 5 °C. Rapid decline of generic E. coli and E. coli O157:H7 populations was observed for all plant types regardless of the leaf age at the time of inoculation and the irrigation type across both seasonal growth cycle trials. The decline rate of the surviving populations for the fall season was slower than for the summer season. The minimal processing with 3 mg/L of ClO₂ was not sufficient to fully disinfect the inoculated leaves prior to packaging and refrigerated storage. Viable populations of E. coli and E. coli O157:H7 were confirmed throughout storage, including the final time point at the end of acceptable visual leaf quality. In this study, the ability of low populations of E. coli to survive during production and postharvest operations in selected mini-greens has been demonstrated. However, further field-based trials are needed to expand understanding of the post-contamination fate of enteric bacterial pathogens on leafy vegetables. In summary, this research work provides baseline data upon which to develop food safety preventive control guidance during the production and minimal processing of these crops. Copyright © 2011 Elsevier B.V. All rights reserved.

Studying the Cost and Value of Library Services: Final Report. Technical Report APLAB/94-3/1,2,3,4.

ERIC Educational Resources Information Center

Kantor, Paul B.; And Others

This is the final technical report (in three parts) of a 15-month long project to study the costs and value of library functions at five major research libraries. Twenty-one services or service aspects were studied, and numerous measures of the importance or benefit of the service to the users were made. These measures were studied together to lay…

An Inquiry into Testing of Information Retrieval Systems. Comparative Systems Laboratory Final Technical Report, Part III: CSL Related Studies.

ERIC Educational Resources Information Center

Zull, Carolyn Gifford, Ed.; And Others

This third volume of the Comparative Systems Laboratory (CSL) Final Technical Report is a collection of relatively independent studies performed on CSL materials. Covered in this document are studies on: (1) properties of files, including a study of the growth rate of a dictionary of index terms as influenced by number of documents in the file and…

Programs of Study as a State Policy Mandate: A Longitudinal Study of the South Carolina Personal Pathways to Success Initiative. Unabridged Final Technical Report (Years 1-5)

ERIC Educational Resources Information Center

Hammond, Cathy; Withington, Cairen; Sharp, Julia L.; Mobley, Catherine; Drew, Sam F.; Stringfield, Samuel C.; Stipanovic, Natalie; Swiger, Caroline M.; Daugherty, Lindsay; Griffith, Cathy

2014-01-01

This final report presents findings from data collection and analysis conducted during a five-year study by the National Dropout Prevention Center (NDPC) at Clemson University, in conjunction with colleagues from the National Research Center for Career and Technical Education (NRCCTE) at the University of Louisville. This project was one of three…

Ground-Based Radiometric Measurements of Slant Path Attenuation in the V/W Bands

DTIC Science & Technology

2016-04-01

GROUND-BASED RADIOMETRIC MEASUREMENTS OF SLANT PATH ATTENUATION IN THE V/W BANDS APRIL 2016 FINAL TECHNICAL REPORT APPROVED FOR PUBLIC RELEASE...2. REPORT TYPE FINAL TECHNICAL REPORT 3. DATES COVERED (From - To) OCT 2012 – SEP 2015 4. TITLE AND SUBTITLE GROUND-BASED RADIOMETRIC MEASUREMENTS ...SUPPLEMENTARY NOTES 14. ABSTRACT Ground-based radiometric techniques were applied to measure the slant path attenuation cumulative distribution function to

Final Technical Progress Report: Development of Low-Cost Suspension Heliostat; December 7, 2011 - December 6, 2012

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bender, W.

2013-01-01

Final technical progress report of SunShot Incubator Solaflect Energy. The project succeeded in demonstrating that the Solaflect Suspension Heliostat design is viable for large-scale CSP installations. Canting accuracy is acceptable and is continually improving as Solaflect improves its understanding of this design. Cost reduction initiatives were successful, and there are still many opportunities for further development and further cost reduction.

Frequent combination of antimicrobial multiresistance and extraintestinal pathogenicity in Escherichia coli isolates from urban rats (Rattus norvegicus) in Berlin, Germany.

PubMed

Guenther, Sebastian; Bethe, Astrid; Fruth, Angelika; Semmler, Torsten; Ulrich, Rainer G; Wieler, Lothar H; Ewers, Christa

2012-01-01

Urban rats present a global public health concern as they are considered a reservoir and vector of zoonotic pathogens, including Escherichia coli. In view of the increasing emergence of antimicrobial resistant E. coli strains and the on-going discussion about environmental reservoirs, we intended to analyse whether urban rats might be a potential source of putatively zoonotic E. coli combining resistance and virulence. For that, we took fecal samples from 87 brown rats (Rattus norvegicus) and tested at least three E. coli colonies from each animal. Thirty two of these E. coli strains were pre-selected from a total of 211 non-duplicate isolates based on their phenotypic resistance to at least three antimicrobial classes, thus fulfilling the definition of multiresistance. As determined by multilocus sequence typing (MLST), these 32 strains belonged to 24 different sequence types (STs), indicating a high phylogenetic diversity. We identified STs, which frequently occur among extraintestinal pathogenic E. coli (ExPEC), such as STs 95, 131, 70, 428, and 127. Also, the detection of a number of typical virulence genes confirmed that the rats tested carried ExPEC-like strains. In particular, the finding of an Extended-spectrum beta-lactamase (ESBL)-producing strain which belongs to a highly virulent, so far mainly human- and avian-restricted ExPEC lineage (ST95), which expresses a serogroup linked with invasive strains (O18:NM:K1), and finally, which produces an ESBL-type frequently identified among human strains (CTX-M-9), pointed towards the important role, urban rats might play in the transmission of multiresistant and virulent E. coli strains. Indeed, using a chicken infection model, this strain showed a high in vivo pathogenicity. Imagining the high numbers of urban rats living worldwide, the way to the transmission of putatively zoonotic, multiresistant, and virulent strains might not be far ahead. The unforeseeable consequences of such an emerging public health threat need careful consideration in the future.

Advanced Transportation System Studies Technical Area 2 (TA-2) Heavy Lift Launch Vehicle Development Contract. Volume 2; Technical Results

NASA Technical Reports Server (NTRS)

1995-01-01

The purpose of the Advanced Transportation System Studies (ATSS) Technical Area 2 (TA-2) Heavy Lift Launch Vehicle Development contract was to provide advanced launch vehicle concept definition and analysis to assist NASA in the identification of future launch vehicle requirements. Contracted analysis activities included vehicle sizing and performance analysis, subsystem concept definition, propulsion subsystem definition (foreign and domestic), ground operations and facilities analysis, and life cycle cost estimation. This document is Volume 2 of the final report for the contract. It provides documentation of selected technical results from various TA-2 analysis activities, including a detailed narrative description of the SSTO concept assessment results, a user's guide for the associated SSTO sizing tools, an SSTO turnaround assessment report, an executive summary of the ground operations assessments performed during the first year of the contract, a configuration-independent vehicle health management system requirements report, a copy of all major TA-2 contract presentations, a copy of the FLO launch vehicle final report, and references to Pratt & Whitney's TA-2 sponsored final reports regarding the identification of Russian main propulsion technologies.

Technical Evaluation Motor No. 7 (TEM-7)

NASA Technical Reports Server (NTRS)

Hughes, Phil

1991-01-01

The Technical Evaluation Motor No. 7 (TEM-7) test was a full-scale, full duration static test firing of a high performance motor-configuration solid rocket motor with nozzle vectoring. The final test report documents the procedures, performance, and results of the static test firing of TEM-7. All observations, discussions, conclusions, and recommendations included in the report are complete and final except for the TEM-7 fixed housing unbond investigation. A presentation and discussion of TEM-7 performance, anomalies, and test result concurrence with the objectives outlined in CTP-0107, Rev A, Space Shuttle Technical Evaluation Motor No. 7 (TEM-7) Static Fire Test Plan are included.

Building Stronger State Energy Partnerships with the U.S. Department of Energy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Marks, Kate

2011-09-30

This final technical report details the results of total work efforts and progress made from October 2007 – September 2011 under the National Association of State Energy Officials (NASEO) cooperative agreement DE-FC26-07NT43264, Building Stronger State Energy Partnerships with the U.S. Department of Energy. Major topical project areas in this final report include work efforts in the following areas: Energy Assurance and Critical Infrastructure, State and Regional Technical Assistance, Regional Initiative, Regional Coordination and Technical Assistance, and International Activities in China. All required deliverables have been provided to the National Energy Technology Laboratory and DOE program officials.

Final Technical Report of Project DE-FG02-96ER14647

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lundeen, Stephen R.

This is the final technical report of work completed under DOE support over the period Sept. 1, 1996 until May 31, 2015. The title of the project was "Ion/Excited Atom Collision Studies with a Rydberg Target and a CO2 Laser" from 9/1/96 to 10/31/06, and "Properties of Actinide Ions from Measurements of Rydberg Ion Fine Structure" from 11/1/06 until 5/31/15. The primary technical results were a detailed experimental study of resonant charge transfer between Rydberg atoms and highly-charged ions, and unique measurements of many properties of multiply-charged Thorium ions.

AN ESCHERICHIA COLI STRAIN GENOMICALLY MODIFIED TO FACILITATE ENVIRONMENTAL DETECTION AND STUDY. (R825354)

EPA Science Inventory

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

BACTERIOPHAGE T4 MULTIPLICATION IN A GLUCOSE-LIMITED ESCHERICHIA COLI BIOFILM. (R825503)

EPA Science Inventory

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

In-Situ Biocatalytic Production of Trehalose with Autoinduction Expression of Trehalose Synthase.

PubMed

Yan, Xincheng; Zhu, Liying; Yu, Yadong; Xu, Qing; Huang, He; Jiang, Ling

2018-02-14

We developed an in-situ biocatalytic process that couples autoinduction expression of trehalose synthase (TreS) and whole-cell catalysis for trehalose production. With lactose as the autoinducer, the activity of recombinant TreS in recombinant Escherichia coli was optimized through a visualization method, which resulted in a maximum value of 12 033 ± 730 U/mL in pH-stat fed-batch fermentation mode. Meanwhile, the permeability of the autoinduced E. coli increased significantly, which makes it possible to be directly used as a whole-cell biocatalyst for trehalose production, whereby the byproduct glucose can also act as an extra carbon source. In this case, the final yield of trehalose was improved to 90.5 ± 5.7% and remained as high as 83.2 ± 5.0% at the 10th batch, which is the highest value achieved using recombinant TreS. Finally, an integrated strategy for trehalose production was established, and its advantages compared to the traditional mode have been summarized.

The diversity of beef safety: A global reason to strengthen our current systems.

PubMed

Brashears, Mindy M; Chaves, Byron D

2017-10-01

The purpose of this paper is to propose a more integrated and more aggressive system approach to food safety rather than focusing on one segment of the industry, or on one approach as described by or constrained by one set of regulations. We focus on the prevalence and control measures for Salmonella and pathogenic Escherichia coli, particularly, Shiga toxin-producing E. coli (STEC) in live cattle on the farm and in the final raw beef product at retail. We describe the antimicrobial and process control strategies most commonly used during slaughter and processing to prevent and reduce the frequency and concentration of these pathogens in the final product, and we propose points along the food chain where more interventions can be applied to ultimately reduce the prevalence of foodborne pathogens associated with beef and beef products, and to protect public health as well the global food supply. Copyright © 2017 Elsevier Ltd. All rights reserved.

Utilization of alkaline phosphatase PhoA in the bioproduction of geraniol by metabolically engineered Escherichia coli.

PubMed

Liu, Wei; Zhang, Rubing; Tian, Ning; Xu, Xin; Cao, Yujing; Xian, Mo; Liu, Huizhou

2015-01-01

Geraniol is a valuable acyclic monoterpene alcohol and has many applications in the perfume industries, pharmacy and others. It has been hypothesized that phosphatases can convert geranyl diphosphate (GPP) into geraniol. However, whether and which phosphatases can transform GPP to geraniol has remained unanswered up till now. In this paper, the catalysis abilities of 4 different types of phosphatases were studied with GPP as substrate in vitro. They are bifunctional diacylglycerol diphosphate phosphatase (DPP1) and lipid phosphate phosphatase (LPP1) from Saccharomyces cerevisiae, ADP-ribose pyrophosphatase (NudF) and alkaline phosphatase (PhoA) from Escherichia coli. The results show that just PhoA from E. coli can convert GPP into geraniol. Moreover, in order to confirm the ability of PhoA in vivo, the heterologous mevalonate pathway and geranyl diphosphate synthase gene from Abies grandis were co-overexpressed in E. coli with PhoA gene and 5.3 ± 0.2 mg/l geraniol was produced from glucose in flask-culture. Finally, we also evaluated the fed-batch fermentation of this engineered E. coli and a maximum concentration of 99.3 mg/l geraniol was produced while the conversion efficiency of glucose to geranoid (gram to gram) was 0.51%. Our results offer a new option for geraniol biosynthesis and promote the industrial bio-production of geraniol.

Utilization of alkaline phosphatase PhoA in the bioproduction of geraniol by metabolically engineered Escherichia coli

PubMed Central

Liu, Wei; Zhang, Rubing; Tian, Ning; Xu, Xin; Cao, Yujing; Xian, Mo; Liu, Huizhou

2015-01-01

Geraniol is a valuable acyclic monoterpene alcohol and has many applications in the perfume industries, pharmacy and others. It has been hypothesized that phosphatases can convert geranyl diphosphate (GPP) into geraniol. However, whether and which phosphatases can transform GPP to geraniol has remained unanswered up till now. In this paper, the catalysis abilities of 4 different types of phosphatases were studied with GPP as substrate in vitro. They are bifunctional diacylglycerol diphosphate phosphatase (DPP1) and lipid phosphate phosphatase (LPP1) from Saccharomyces cerevisiae, ADP-ribose pyrophosphatase (NudF) and alkaline phosphatase (PhoA) from Escherichia coli. The results show that just PhoA from E. coli can convert GPP into geraniol. Moreover, in order to confirm the ability of PhoA in vivo, the heterologous mevalonate pathway and geranyl diphosphate synthase gene from Abies grandis were co-overexpressed in E. coli with PhoA gene and 5.3 ± 0.2 mg/l geraniol was produced from glucose in flask-culture. Finally, we also evaluated the fed-batch fermentation of this engineered E. coli and a maximum concentration of 99.3 mg/l geraniol was produced while the conversion efficiency of glucose to geranoid (gram to gram) was 0.51%. Our results offer a new option for geraniol biosynthesis and promote the industrial bio-production of geraniol. PMID:26091008

Label-free ITO-based immunosensor for the detection of very low concentrations of pathogenic bacteria.

PubMed

Barreiros dos Santos, M; Azevedo, S; Agusil, J P; Prieto-Simón, B; Sporer, C; Torrents, E; Juárez, A; Teixeira, V; Samitier, J

2015-02-01

Here we describe the fabrication of a highly sensitive and label-free ITO-based impedimetric immunosensor for the detection of pathogenic bacteria Escherichia coli O157:H7. Anti-E. coli antibodies were immobilized onto ITO electrodes using a simple, robust and direct methodology. First, the covalent attachment of epoxysilane on the ITO surface was demonstrated by Atomic Force Microscopy and cyclic voltammetry. The immobilization of antibody on the epoxysilane layer was quantified by Optical Waveguide Lightmode Spectroscopy, obtaining a mass variation of 12 ng cm(− 2) (0.08 pmol cm(− 2)). Microcontact printing and fluorescence microscopy were used to demonstrate the specific binding of E. coli O157:H7 to the antibody-patterned surface. We achieved a ratio of 1:500 Salmonella typhimurium/E. coli O157:H7, thus confirming the selectivity of the antibodies and efficiency of the functionalization procedure. Finally, the detection capacity of the ITO-based immunosensor was evaluated by Electrochemical Impedance Spectroscopy. A very low limit of detection was obtained (1 CFU mL(− 1)) over a large linear working range (10–10(6) CFU mL(− 1)). The specificity of the impedimetric immunosensor was also examined. Less than 20% of non-specific bacteria (S. typhimurium and E. coli K12) was observed. Our results reveal the applicability of ITO for the development of highly sensitive and selective impedimetric immunosensors.

Preparation of immunomagnetic iron-dextran nanoparticles and application in rapid isolation of E.coli O157:H7 from foods

PubMed Central

Duan, Hui-Li; Shen, Zhi-Qiang; Wang, Xin-Wei; Chao, Fu-Huan; Li, Jun-Wen

2005-01-01

AIM: To prepare a kind of magnetic iron-dextran nanoparticles that was coated with anti-E.coli O157:H7 IgG, analyze its application conditions, and try to use it to isolate E.coli O157:H7 from foods. METHODS: Magnetic iron-dextran nanoparticles were prepared by the reaction of a mixture of ferric and ferrous ions with dextran polymers under alkaline conditions. The particles were coated with antiserum against E.coli O157:H7 by the periodate oxidation-borohydride reduction procedure. The oxidation time, amount of antibody coating the particles, amount of nanoparticles, incubation time and isolation time were varied to determine their effects on recovery of the organisms. Finally, the optimum conditions for isolating E.coli O157:H7 from food samples were established. RESULTS: E.coli O157:H7 can be isolated from samples within 15 min with the sensitivity of 101 CFU/mL or even less. In the presence of 108 CFU/mL of other organisms, the sensitivity is 101-102 CFU/mL. Nonspecific binding of other bacteria to the particles was not observed. Two and a half hours of enrichment is enough for the particles to detect the target from the food samples inoculated with 1 CFU/g. CONCLUSION: Isolation of target bacteria by immuno-magnetic nanoparticles is an efficient method with high sensitivity and specificity. The technique is so simple that it can be operated in lab and field even by untrained personnel. PMID:15968716

Computer-aided teniae coli detection using height maps from computed tomographic colonography images

NASA Astrophysics Data System (ADS)

Wei, Zhuoshi; Yao, Jianhua; Wang, Shijun; Summers, Ronald M.

2011-03-01

Computed tomographic colonography (CTC) is a minimally invasive technique for colonic polyps and cancer screening. Teniae coli are three bands of longitudinal smooth muscle on the colon surface. They are parallel, equally distributed on the colon wall, and form a triple helix structure from the appendix to the sigmoid colon. Because of their characteristics, teniae coli are important anatomical meaningful landmarks on human colon. This paper proposes a novel method for teniae coli detection on CT colonography. We first unfold the three-dimensional (3D) colon using a reversible projection technique and compute the two-dimensional (2D) height map of the unfolded colon. The height map records the elevation of colon surface relative to the unfolding plane, where haustral folds corresponding to high elevation points and teniae to low elevation points. The teniae coli are detected on the height map and then projected back to the 3D colon. Since teniae are located where the haustral folds meet, we break down the problem by first detecting haustral folds. We apply 2D Gabor filter banks to extract fold features. The maximum response of the filter banks is then selected as the feature image. The fold centers are then identified based on piecewise thresholding on the feature image. Connecting the fold centers yields a path of the folds. Teniae coli are finally extracted as lines running between the fold paths. Experiments were carried out on 7 cases. The proposed method yielded a promising result with an average normalized RMSE of 5.66% and standard deviation of 4.79% of the circumference of the colon.

Overexpression of biotin synthase and biotin ligase is required for efficient generation of sulfur-35 labeled biotin in E. coli.

PubMed

Delli-Bovi, Teegan A; Spalding, Maroya D; Prigge, Sean T

2010-10-11

Biotin is an essential enzyme cofactor that acts as a CO2 carrier in carboxylation and decarboxylation reactions. The E. coli genome encodes a biosynthetic pathway that produces biotin from pimeloyl-CoA in four enzymatic steps. The final step, insertion of sulfur into desthiobiotin to form biotin, is catalyzed by the biotin synthase, BioB. A dedicated biotin ligase (BirA) catalyzes the covalent attachment of biotin to biotin-dependent enzymes. Isotopic labeling has been a valuable tool for probing the details of the biosynthetic process and assaying the activity of biotin-dependent enzymes, however there is currently no established method for 35S labeling of biotin. In this study, we produced [35S]-biotin from Na35SO4 and desthiobiotin with a specific activity of 30.7 Ci/mmol, two orders of magnitude higher than previously published methods. The biotinylation domain (PfBCCP-79) from the Plasmodium falciparum acetyl-CoA carboxylase (ACC) was expressed in E. coli as a biotinylation substrate. We found that overexpression of the E. coli biotin synthase, BioB, and biotin ligase, BirA, increased PfBCCP-79 biotinylation 160-fold over basal levels. Biotinylated PfBCCP-79 was purified by affinity chromatography, and free biotin was liberated using acid hydrolysis. We verified that we had produced radiolabeled biologically active [D]-biotin that specifically labels biotinylated proteins through reuptake in E. coli. The strategy described in our report provides a simple and effective method for the production of [35S]-biotin in E. coli based on affinity chromatography.

78 FR 34392 - Guidance for Industry and Food and Drug Administration Staff: Technical Considerations for Pen...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-07

...] Guidance for Industry and Food and Drug Administration Staff: Technical Considerations for Pen, Jet, and... availability of a final guidance document entitled ``Technical Considerations for Pen, Jet, and Related... developing information to support a marketing application for a pen, jet, or related injector device intended...

Florida Study of Career and Technical Education. Final Report

ERIC Educational Resources Information Center

Jacobson, Louis; Mokher, Christine

2014-01-01

A key goal of the "Carl D. Perkins Career and Technical Education Act of 2006" ("Perkins IV") is to ensure career and technical education (CTE) programs are widely available for preparing high school and college students for "high skill, high wage, or high demand occupations in current or emerging professions"…

Application of the Modular Approach to an In-House Validation Study of Real-Time PCR Methods for the Detection and Serogroup Determination of Verocytotoxigenic Escherichia coli ▿ †

PubMed Central

Kagkli, Dafni-Maria; Weber, Thomas P.; Van den Bulcke, Marc; Folloni, Silvia; Tozzoli, Rosangela; Morabito, Stefano; Ermolli, Monica; Gribaldo, Laura; Van den Eede, Guy

2011-01-01

European Commission regulation 2073/2005 on the microbiological criteria for food requires that Escherichia coli is monitored as an indicator of hygienic conditions. Since verocytotoxigenic E. coli (VTEC) strains often cause food-borne infections by the consumption of raw food, the Biological Hazards (BIOHAZ) panel of the European Food Safety Authority (EFSA) recommended their monitoring in food as well. In particular, VTEC strains belonging to serogroups such as O26, O103, O111, O145, and O157 are known causative agents of several human outbreaks. Eight real-time PCR methods for the detection of E. coli toxin genes and their variants (stx1, stx2), the intimin gene (eae), and five serogroup-specific genes have been proposed by the European Reference Laboratory for VTEC (EURL-VTEC) as a technical specification to the European Normalization Committee (CEN TC275/WG6). Here we applied a “modular approach” to the in-house validation of these PCR methods. The modular approach subdivides an analytical process into separate parts called “modules,” which are independently validated based on method performance criteria for a limited set of critical parameters. For the VTEC real-time PCR module, the following parameters are being assessed: specificity, dynamic range, PCR efficiency, and limit of detection (LOD). This study describes the modular approach for the validation of PCR methods to be used in food microbiology, using single-target plasmids as positive controls and showing their applicability with food matrices. PMID:21856838

Systematic Engineering of Escherichia coli for d-Lactate Production from Crude Glycerol.

PubMed

Wang, Zei Wen; Saini, Mukesh; Lin, Li-Jen; Chiang, Chung-Jen; Chao, Yun-Peng

2015-11-04

Crude glycerol resulting from biodiesel production is an abundant and renewable resource. However, the impurities in crude glycerol usually make microbial fermentation problematic. This issue was addressed by systematic engineering of Escherichia coli for the production of d-lactate from crude glycerol. First, mgsA and the synthetic pathways of undesired products were eliminated in E. coli, rendering the strain capable of homofermentative production of optically pure d-lactate. To direct carbon flux toward d-lactate, the resulting strain was endowed with an enhanced expression of glpD-glpK in the glycerol catabolism and of a heterologous gene encoding d-lactate dehydrogenase. Moreover, the strain was evolved to improve its utilization of cruder glycerol and subsequently equipped with the FocA channel to export intracellular d-lactate. Finally, the fed-batch fermentation with two-phase culturing was carried out with a bioreactor. As a result, the engineered strain enabled production of 105 g/L d-lactate (99.9% optical purity) from 121 g/L crude glycerol at 40 h. The result indicates the feasibility of our approach to engineering E. coli for the crude glycerol-based fermentation.

Hydrothermal green synthesis of magnetic Fe3O4-carbon dots by lemon and grape fruit extracts and as a photoluminescence sensor for detecting of E. coli bacteria.

PubMed

Ahmadian-Fard-Fini, Shahla; Salavati-Niasari, Masoud; Ghanbari, Davood

2018-10-05

The aim of this work is preparing of a photoluminescence nanostructures for rapid detection of bacterial pathogens. Firstly, carbon dots (CDs) were synthesized by grape fruit, lemon, turmeric extracts and hydrothermal method. Then Fe 3 O 4 (magnetite) nanoparticles was achieved using these bio-compatible capping agents. Finally, magnetite-carbon dots were synthesized as a novel magnetic and photoluminescence nanocomposite. X-ray diffraction (XRD) confirms the crystallinity and phase of the products, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) investigate the morphology, shape and size of the magnetite, carbon dot and nanocomposites. Fourier transform infrared (FT-IR) spectroscopy shows the purity of the nanostructures. Ultraviolet-visible (UV-Vis) absorption and photo-luminescence (PL) spectroscopy show suitable photo-luminescence under ultraviolet irradiation. Vibrating sample magnetometer (VSM) shows super paramagnetic property of the product. Interestingly carbon dots were used as a non-toxic photoluminescence sensor for detecting of Escherichia coli (E. coli) bacteria. Results show quenching of photoluminescence of the CDs nanocomposite by increasing amount of E. coli bacteria. Copyright © 2018 Elsevier B.V. All rights reserved.

Risk factors for Escherichia coli O157 shedding and super-shedding by dairy heifers at pasture.

PubMed

Williams, K J; Ward, M P; Dhungyel, O P; Hall, E J S

2015-04-01

We undertook a longitudinal study within a cohort of 52 dairy heifers maintained under constant management systems and sampled weekly to investigate a comprehensive range of risk factors which may influence shedding or super-shedding of E. coli O157 (detected by direct faecal culture and immunomagnetic separation). E. coli O157 was detected from 416/933 (44.6%) samples (faeces and recto-anal mucosal swabs) and 32 (3.4%) samples enumerated at >10000 c.f.u./g. Weekly point prevalence ranged from 9.4% to 94.3%. Higher temperature (P < 0.001), rainfall (P = 0.02), relative humidity (P < 0.001), pasture growth (P = 0.013) and body score (P = 0.029) were positively associated with increased shedding. Higher rainfall (P < 0.001), hide contamination (P = 0.002) and increased faecal consistency (P = 0.023) were positively associated with super-shedding. Increased solar exposure had a negative effect on both shedding and super-shedding within bivariate analyses but in the final multivariate model for shedding demonstrated a positive effect (P = 0.017). Results suggest that environmental factors are important in E. coli O157 shedding in cattle.

Detoxifying Escherichia coli for endotoxin-free production of recombinant proteins.

PubMed

Mamat, Uwe; Wilke, Kathleen; Bramhill, David; Schromm, Andra Beate; Lindner, Buko; Kohl, Thomas Andreas; Corchero, José Luis; Villaverde, Antonio; Schaffer, Lana; Head, Steven Robert; Souvignier, Chad; Meredith, Timothy Charles; Woodard, Ronald Wesley

2015-04-16

Lipopolysaccharide (LPS), also referred to as endotoxin, is the major constituent of the outer leaflet of the outer membrane of virtually all Gram-negative bacteria. The lipid A moiety, which anchors the LPS molecule to the outer membrane, acts as a potent agonist for Toll-like receptor 4/myeloid differentiation factor 2-mediated pro-inflammatory activity in mammals and, thus, represents the endotoxic principle of LPS. Recombinant proteins, commonly manufactured in Escherichia coli, are generally contaminated with endotoxin. Removal of bacterial endotoxin from recombinant therapeutic proteins is a challenging and expensive process that has been necessary to ensure the safety of the final product. As an alternative strategy for common endotoxin removal methods, we have developed a series of E. coli strains that are able to grow and express recombinant proteins with the endotoxin precursor lipid IVA as the only LPS-related molecule in their outer membranes. Lipid IVA does not trigger an endotoxic response in humans typical of bacterial LPS chemotypes. Hence the engineered cells themselves, and the purified proteins expressed within these cells display extremely low endotoxin levels. This paper describes the preparation and characterization of endotoxin-free E. coli strains, and demonstrates the direct production of recombinant proteins with negligible endotoxin contamination.

Engineering a Synthetic Dual-Organism System for Hydrogen Production▿

PubMed Central

Waks, Zeev; Silver, Pamela A.

2009-01-01

Molecular hydrogen produced biologically from renewable biomass is an attractive replacement for fossil fuels. One potential route for biological hydrogen production is the conversion of biomass into formate, which can subsequently be processed into hydrogen by Escherichia coli. Formate is also a widely used commodity chemical, making its bioproduction even more attractive. Here we demonstrate the implementation of a formate-overproducing pathway in Saccharomyces cerevisiae, a well-established industrial organism. By expressing the anaerobic enzyme pyruvate formate lyase from E. coli, we engineered a strain of yeast that overproduced formate relative to undetectable levels in the wild type. The addition of a downstream enzyme, AdhE of E. coli, resulted in an additional 4.5-fold formate production increase as well as an increase in growth rate and biomass yield. Overall, an 18-fold formate increase was achieved in a strain background whose formate degradation pathway had been deleted. Finally, as a proof of concept, we were able to produce hydrogen from this formate-containing medium by using E. coli as a catalyst in a two-step process. With further optimizations, it may be feasible to use S. cerevisiae on a larger scale as the foundation for yeast-based biohydrogen. PMID:19201964

A cellulosic responsive "living" membrane.

PubMed

Qin, Guokui; Panilaitis, Bruce J; Kaplan, Zhongyuan Sun David L

2014-01-16

Bacterial cellulose has been demonstrated to be a remarkably versatile biomaterial and widely used in biomedical applications due to its unique physical properties. Here we reported for the first time a "living membrane" system based on recombinant Escherichia coli bacterial strains entrapped in cellulosic membranes produced by Gluconacetobacter xylinus. Biologically driven detection and identification of a range of target molecules presents unique challenges, and requires that detection methods are developed to be rapid, specific and sensitive. The compatibility of G. xylinus and recombinant E. coli strains was first investigated for co-cultivation, and the relationship between the number of entrapped E. coli and the level of inducible signal achieved was further explored by fluorescent signal observation in confocal microscopy. Finally to amplify the response to inducers for maximum fluorescent signal, a positive-feedback genetic amplifier was designed within recombinant E. coli strain entrapped in the living cellulosic membrane system, allowing for the detection mechanism to be extremely sensitive and resulting in a significant fluorescent signal from a single receptor binding event. The living membrane system proposed here will create devices of greater complexity in function for applications in biological and chemical detection. Copyright © 2013. Published by Elsevier Ltd.

Final Technical Report for Award DESC0011912, "Trimodal Tapping Mode Atomic Force Microscopy: Simultaneous 4D Mapping of Conservative and Dissipative Probe-Sample Interactions of Energy-Relevant Materials”

DOE Office of Scientific and Technical Information (OSTI.GOV)

Solares, Santiago D.

The final project report covering the period 7/1/14-6/30/17 provides an overview of the technical accomplishments in the areas of (i) fundamental viscoelasticity, (ii) multifrequency atomic force microscopy, and (iii) characterization of energy-relevant materials with atomic force microscopy. A list of publications supported by the project is also provided.

Application of Knowledge-Based Techniques to Tracking Function

DTIC Science & Technology

2006-09-01

38394041 42434445 46474849 505152 53545556 57585960 616263 646566 676869 707172 737475 7677 7879 8081 8283 8485 8687 8889 9091 9293 9495 969798 99100...Knowledge-based applications to adaptive space-time processing. Volume I: Summary”, AFRL-SN-TR-2001-146 Vol. I (of Vol. VI ), Final Technical Report, July...2001-146 Vol. IV (of Vol. VI ), Final Technical Report, July 2001. [53] C. Morgan, L. Moyer, “Knowledge-based applications to adaptive space-time

Forecasting the Future Food Service World of Work. Final Report. Volume III. Technical Papers on the Future of the Food Service Industry. Service Management Reports.

ERIC Educational Resources Information Center

Powers, Thomas F., Ed.; Swinton, John R., Ed.

This third and final volume of a study on the future of the food service industry contains the technical papers on which the information in the previous two volumes was based. The papers were written by various members of the Pennsylvania State University departments of economics, food science, nutrition, social psychology, and engineering and by…

Final Technical Report for contract number DE-FG02-05ER15670

DOE Office of Scientific and Technical Information (OSTI.GOV)

Glazebrook, Jane

This is the final technical report for contract number DE-FG02-05ER15670. The project is now complete, and results of the project have been published. Two papers were published based on work done in the last three-year funding period. The DOIs of these papers are included below. The abstracts of the papers, providing summaries of the work, are included in the body of the report.

Astrobee Periodic Technical Review (PTR) Delta 3

NASA Technical Reports Server (NTRS)

Provencher, Christopher; Smith, Marion F.; Smith, Ernest Everett; Bualat, Maria Gabriele; Barlow, Jonathan Spencer

2017-01-01

Astrobee is a free flying robot for the inside of the International Space Station (ISS). The Periodic Technical Review (PTR) delta 3 is the final design review of the system presented to stakeholders.

Polar source analysis : technical memorandum

DOT National Transportation Integrated Search

2017-09-29

The following technical memorandum describes the development, testing and analysis of various polar source data sets. The memorandum also includes recommendation for potential inclusion in future releases of AEDT. This memorandum is the final deliver...

2005 v4.2 Technical Support Document

EPA Pesticide Factsheets

Technical Support Document for the Final Transport Rule describes how updated 2005 NEI, version 2 emissions and were processed for air quality modeling in support of the Cross-state Air Pollution Rule (CSAPR).

Direct Final Rule for Technical Amendments for Marine Spark-Ignition Engines and Vessels

EPA Pesticide Factsheets

Rule published September 16, 2010 to make technical amendments to the design standard for portable marine fuel tanks. This rule incorporates safe recommended practices, developed through industry consensus.

On-farm Campylobacter and Escherichia coli in commercial broiler chickens: Re-used bedding does not influence Campylobacter emergence and levels across sequential farming cycles

PubMed Central

Chinivasagam, H. N.; Estella, W.; Rodrigues, H.; Mayer, D. G.; Weyand, C.; Tran, T.; Onysk, A.; Diallo, I.

2016-01-01

Limitations in quality bedding material have resulted in the growing need to re-use litter during broiler farming in some countries, which can be of concern from a food-safety perspective. The aim of this study was to compare the Campylobacter levels in ceca and litter across three litter treatments under commercial farming conditions. The litter treatments were (a) the use of new litter after each farming cycle; (b) an Australian partial litter re-use practice; and (c) a full litter re-use practice. The study was carried out on two farms over two years (Farm 1, from 2009–2010 and Farm 2, from 2010–2011), across three sheds (35,000 to 40,000 chickens/shed) on each farm, adopting three different litter treatments across six commercial cycles. A random sampling design was adopted to test litter and ceca for Campylobacter and Escherichia coli, prior to commercial first thin-out and final pick-up. Campylobacter levels varied little across litter practices and farming cycles on each farm and were in the range of log 8.0–9.0 CFU/g in ceca and log 4.0–6.0 MPN/g for litter. Similarly the E. coli in ceca were ∼log 7.0 CFU/g. At first thin-out and final pick-up, the statistical analysis for both litter and ceca showed that the three-way interaction (treatments by farms by times) was highly significant (P < 0.01), indicating that the patterns of Campylobacter emergence/presence across time vary between the farms, cycles and pickups. The emergence and levels of both organisms were not influenced by litter treatments across the six farming cycles on both farms. Either C. jejuni or C. coli could be the dominant species across litter and ceca, and this phenomenon could not be attributed to specific litter treatments. Irrespective of the litter treatments in place, cycle 2 on Farm 2 remained Campylobacter-free. These outcomes suggest that litter treatments did not directly influence the time of emergence and levels of Campylobacter and E. coli during commercial farming. PMID:26908887

5S RRNA GENE DELETIONS CAUSE AN UNEXPECTEDLY HIGH FITNESS LOSS IN ESCHERICHIA COLI. (R825354)

EPA Science Inventory

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

RNase H-assisted RNA-primed rolling circle amplification for targeted RNA sequence detection.

PubMed

Takahashi, Hirokazu; Ohkawachi, Masahiko; Horio, Kyohei; Kobori, Toshiro; Aki, Tsunehiro; Matsumura, Yukihiko; Nakashimada, Yutaka; Okamura, Yoshiko

2018-05-17

RNA-primed rolling circle amplification (RPRCA) is a useful laboratory method for RNA detection; however, the detection of RNA is limited by the lack of information on 3'-terminal sequences. We uncovered that conventional RPRCA using pre-circularized probes could potentially detect the internal sequence of target RNA molecules in combination with RNase H. However, the specificity for mRNA detection was low, presumably due to non-specific hybridization of non-target RNA with the circular probe. To overcome this technical problem, we developed a method for detecting a sequence of interest in target RNA molecules via RNase H-assisted RPRCA using padlocked probes. When padlock probes are hybridized to the target RNA molecule, they are converted to the circular form by SplintR ligase. Subsequently, RNase H creates nick sites only in the hybridized RNA sequence, and single-stranded DNA is finally synthesized from the nick site by phi29 DNA polymerase. This method could specifically detect at least 10 fmol of the target RNA molecule without reverse transcription. Moreover, this method detected GFP mRNA present in 10 ng of total RNA isolated from Escherichia coli without background DNA amplification. Therefore, this method can potentially detect almost all types of RNA molecules without reverse transcription and reveal full-length sequence information.

Seattle To Portland Inter-City ITS Corridor Study And Communications Plan, Final Report

DOT National Transportation Integrated Search

1996-03-01

THIS DOCUMENT IS THE FINAL REPORT PRESENTING THE SEATTLE TO PORTLAND INTELLIGENT TRANSPORTATION SYSTEM (ITS) EARLY DEPLOYMENT PLAN. THE FINAL REPORT SYNTHESIZES INFORMATION FROM TECHNICAL MEMORANDUMS 1 THROUGH 5; INCLUDING EXISTING AND FUTURE CONDITI...

Final Environmental Impact Statement (FEIS)/Final Environmental Impact Report (FEIR). Otis Air National Guard Base, Wastewater Treatment Facility

DTIC Science & Technology

1990-06-01

and G.E. Ness, 1982, Survival of Vibrio cholerae and Escherichia coli in Estuarine Water and Sediments, Applied and Environmental Microbiology, 43...and publications in areas of water and wastewater treatment. David Tomasko, Ph.D., 1985. University of New Mexico . Staff Hydrogeologist. Research...Reserve in California. We are working on EIS’s for the U.S. Air Force Base Closings in realignment in California, New Mexico and Washington, and a very

Benchmark Analysis of Career and Technical Education in Lenawee County. Final Report.

ERIC Educational Resources Information Center

Hollenbeck, Kevin

The career and technical education (CTE) provided in grades K-12 in the county's vocational-technical center and 12 local public school districts of Lenawee County, Michigan, was benchmarked with respect to its attention to career development. Data were collected from the following sources: structured interviews with a number of key respondents…

Heterologous Expression of Plant Cell Wall Degrading Enzymes for Effective Production of Cellulosic Biofuels

PubMed Central

Jung, Sang-Kyu; Parisutham, Vinuselvi; Jeong, Seong Hun; Lee, Sung Kuk

2012-01-01

A major technical challenge in the cost-effective production of cellulosic biofuel is the need to lower the cost of plant cell wall degrading enzymes (PCDE), which is required for the production of sugars from biomass. Several competitive, low-cost technologies have been developed to produce PCDE in different host organisms such as Escherichia coli, Zymomonas mobilis, and plant. Selection of an ideal host organism is very important, because each host organism has its own unique features. Synthetic biology-aided tools enable heterologous expression of PCDE in recombinant E. coli or Z. mobilis and allow successful consolidated bioprocessing (CBP) in these microorganisms. In-planta expression provides an opportunity to simplify the process of enzyme production and plant biomass processing and leads to self-deconstruction of plant cell walls. Although the future of currently available technologies is difficult to predict, a complete and viable platform will most likely be available through the integration of the existing approaches with the development of breakthrough technologies. PMID:22911272

Strategies used for genetically modifying bacterial genome: ite-directed mutagenesis, gene inactivation, and gene over-expression*

PubMed Central

Xu, Jian-zhong; Zhang, Wei-guo

2016-01-01

With the availability of the whole genome sequence of Escherichia coli or Corynebacterium glutamicum, strategies for directed DNA manipulation have developed rapidly. DNA manipulation plays an important role in understanding the function of genes and in constructing novel engineering bacteria according to requirement. DNA manipulation involves modifying the autologous genes and expressing the heterogenous genes. Two alternative approaches, using electroporation linear DNA or recombinant suicide plasmid, allow a wide variety of DNA manipulation. However, the over-expression of the desired gene is generally executed via plasmid-mediation. The current review summarizes the common strategies used for genetically modifying E. coli and C. glutamicum genomes, and discusses the technical problem of multi-layered DNA manipulation. Strategies for gene over-expression via integrating into genome are proposed. This review is intended to be an accessible introduction to DNA manipulation within the bacterial genome for novices and a source of the latest experimental information for experienced investigators. PMID:26834010

International Project on Technical and Vocational Education (UNEVOC). International Advisory Committee (Third Session, Paris, France, October 2-4, 1995). Final Report = Projet International pour l'Enseignement Technique et Professionnel (UNEVOC). Comite Consultatif International (Troisieme Session, Paris, France, 2-4 octobre 1995). Rapport Final.

ERIC Educational Resources Information Center

United Nations Educational, Scientific, and Cultural Organization, Paris (France).

The international advisory committee of the International Project on Technical and Vocational Education (UNEVOC) held its third session in Paris in October 1995. Advisory committee members and observers from the United Nations' specialized, intergovernmental, and nongovernmental organizations reviewed the major UNEVOC project activities undertaken…

International Project on Technical and Vocational Education (UNEVOC). International Advisory Committee (Second Session, Paris, France, December 12-14, 1994). Final Report = Projet International pour l'Enseignement Technique et Professionnel (UNEVOC). Comite Consultatif International (Deuxieme Session, Paris, France, 12-14 decembre 1994). Rapport Final.

ERIC Educational Resources Information Center

United Nations Educational, Scientific, and Cultural Organization, Paris (France).

The international advisory committee of the International Project on Technical and Vocational Education (UNEVOC) held its second session in Paris in December 1994. Nine advisory committee members and observers from the United Nations' specialized, intergovernmental, and nongovernmental organizations reviewed the major UNEVOC project activities…

Final Technical Report for ARRA Funding

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rusack, Roger; Mans, Jeremiah; Poling, Ronald

Final technical report of the University of Minnesota experimental high energy physics group for ARRA support. The Cryogenic Dark Matter Experiment (CDMS) used the funds received to construct a new passive shield to protect a high-purity germanium detector located in the Soudan mine in Northern Minnesota from cosmic rays. The BESIII and the CMS groups purchased computing hardware to assemble computer farms for data analysis and to generate large volumes of simulated data for comparison with the data collected.

Making a Difference in the Lives of Infants and Toddlers and Their Families: The Impacts of Early Head Start. Volumes I-III: Final Technical Report [and] Appendixes [and] Local Contributions to Understanding the Programs and Their Impacts.

ERIC Educational Resources Information Center

Love, John M.; Kisker, Ellen Eliason; Ross, Christine M.; Schochet, Peter Z.; Brooks-Gunn, Jeanne; Paulsell, Diane; Boller, Kimberly; Constantine, Jill; Vogel, Cheri; Fuligni, Alison Sidle; Brady-Smith, Christy

Early Head Start was designed in 1994 as a 2-generation program to enhance children's development and health, strengthen family and community partnerships, and support the staff delivering new services to low-income families with pregnant women, infants, or toddlers. This document contains the final technical report, appendixes, and local…

International Project on Technical and Vocational Education (UNEVOC). International Advisory Committee (First Session, Berlin, Germany, September 20-22, 1993). Final Report = Projet International pour l'Enseignement Technique et Professionnel (UNEVOC). Comite Consultatif International (Premiere Session, Berlin, Germany, 20-22 septembre 1993). Rapport Final.

ERIC Educational Resources Information Center

United Nations Educational, Scientific, and Cultural Organization, Paris (France).

The international advisory committee of the International Project on Technical and Vocational Education (UNEVOC) held its first session in Berlin in September 1993. The advisory committee's 10 members and observers from the United Nations' specialized, intergovernmental, and nongovernmental organizations discussed educational policy, links between…

THE STRINGENT RESPONSE GENES RELA AND SPOT ARE IMPORTANT FOR ESCHERICHIA COLI BIOFILMS UNDER SLOW-GROWTH CONDITIONS. (R825503)

EPA Science Inventory

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

INFLUENCE OF METAL IONS, TEMPERATURE, AND POLYMER CONCENTRATION ON THE CONFORMATION OF ESCHERICHIA COLI K1 CAPSULAR POLYSACCHARIDE. (R825503)

EPA Science Inventory

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

Illinois Clean Coal Institute 2005 annual report. Final technical report for the period September 1st, 2004, through August 31, 2005 on projects funded by the Illinois Department of Commerce and Economic Opportunity

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

2005-11-08

This final technical report contains the abstracts and executive summaries of projects funded through the Illinois Clean Coal Institute solicitation entitled 'Request for proposals No. 04-1(ICCI/RFP04-1)'. Support of these projects is by the Office of Coal Development and Department of Commerce and Economic Opportunity. The projects fall into the following categories: advanced coal mining technologies; coal preparation and coal production business practice; management of coal combustion byproducts; commercialization and technology transfer. Final project extensions are also recorded.

Removal of Listeria monocytogenes dual-species biofilms using combined enzyme-benzalkonium chloride treatments.

PubMed

Rodríguez-López, Pedro; Carballo-Justo, Alba; Draper, Lorraine A; Cabo, Marta L

2017-01-01

The effects of pronase (PRN), cellulase (CEL) or DNaseI alone or combined with benzalkonium chloride (BAC) against Listeria monocytogenes-carrying biofilms were assayed. The best removal activity against L. monocytogenes-Escherichia coli biofilms was obtained using DNaseI followed by PRN and CEL. Subsequently, a modified logistic model was used to quantify the combined effects of PRN or DNaseI with BAC. A better BAC performance after PRN compared to DNaseI eradicating L. monocytogenes was observed. In E. coli the effects were the opposite. Finally, effects of DNaseI and DNaseI-BAC treatments were compared against two different L. monocytogenes-carrying biofilms. DNaseI-BAC was more effective against L. monocytogenes when co-cultured with E. coli. Nonetheless, comparing the removal effects after BAC addition, these were higher in mixed-biofilms with Pseudomonas fluorescens. However, a high number of released viable cells was observed after combined treatments. These results open new perspectives of enzymes as an anti-biofilm strategy for environmental pathogen control.

Single channel atmospheric pressure transporting plasma and plasma stream demultiplexing: physical characterization and application to E. coli bacteria inactivation

NASA Astrophysics Data System (ADS)

Valinataj Omran, A.; Sohbatzadeh, F.; Siadati, S. N.; Hosseinzadeh Colagar, A.; Akishev, Y.; Arefi-Khonsari, F.

2017-08-01

In this article, we developed transporting plasma sources that operate at atmospheric pressure. The effect of electrode configuration on plasma transporting was investigated. In order to increase the transporting plasma cross-section, we converted a plasma stream into four plasma channels by a cylindrical housing. Electron excitation and rotational temperatures were estimated using optical emission spectroscopy. Furthermore, the electrical and temporal characteristics of the plasma, discharge power and charge deposition on the target were investigated. The propagation characteristics of single and multi-channel transporting plasma were compared with the same cross-sectional area. Two configurations for multi-channels were designed for this purpose. Escherichia coli bacteria were exposed to the single and multi-channel transporting discharge for different time durations. After exposure, the results indicated that the inactivation zones were significantly increased by a multi-channel transporting plasma. Finally, E. coli inactivation by those plasma apparatuses was compared with that of several standard antimicrobial test discs such as Gentamicin, Tetracycline, Amoxicillin and Cefixime.

Renewable Fuel Standard (RFS2): Final Rule Additional Resources

EPA Pesticide Factsheets

The final rule of fuels and fuel additives: renewable fuel standard program is published on March 26, 2010 and is effective on July 1, 2010. You will find the links to this final rule and technical amendments supporting this rule.

75 FR 48273 - Technical Service Provider Assistance

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-10

... DEPARTMENT OF AGRICULTURE Natural Resources Conservation Service 7 CFR Part 652 RIN 0578-AA48 Technical Service Provider Assistance AGENCY: Natural Resources Conservation Service, United States Department of Agriculture. ACTION: Final rule; Correcting amendment. SUMMARY: The Natural Resources...

Partial Withdrawal and Final Rule for Nonroad Technical Amendments

EPA Pesticide Factsheets

Amendments to the technical hardship provisions under the Transition Program for Equipment Manufacturers related to the Tier 4 standards for nonroad diesel engines, and to the replacement engine exemption generally applicable to new nonroad engines.

Immunization of pregnant cows with Shiga toxin-2 induces high levels of specific colostral antibodies and lactoferrin able to neutralize E. coli O157:H7 pathogenicity.

PubMed

Albanese, Adriana; Sacerdoti, Flavia; Seyahian, E Abril; Amaral, Maria Marta; Fiorentino, Gabriela; Fernandez Brando, Romina; Vilte, Daniel A; Mercado, Elsa C; Palermo, Marina S; Cataldi, Angel; Zotta, Elsa; Ibarra, Cristina

2018-03-20

E. coli O157:H7 is a foodborne pathogen responsible for bloody diarrhea, hemorrhagic colitis and hemolytic uremic syndrome (HUS). The objective of the present work was to evaluate the ability of colostral IgG obtained from Stx2-immunized cows to prevent against E. coli O157:H7 infection and Stx2 cytotoxicity. Hyperimmune colostrum (HC) was obtained from cows intramuscularly immunized with inactivated Stx2 or vehicle for controls. Colostral IgG was purified by affinity chromatography. Specific IgG antibodies against Stx2 and bovine lactoferrin (bLF) levels in HC and the corresponding IgG (HC-IgG/bLF) were determined by ELISA. The protective effects of HC-IgG/bLF against Stx2 cytotoxicity and adhesion of E. coli O157:H7 and its Stx2-negative mutant were analyzed in HCT-8 cells. HC-IgG/bLF prevention against E. coli O157:H7 was studied in human colon and rat colon loops. Protection against a lethal dose of E. coli O157:H7 was evaluated in a weaned mice model. HC-IgG/bLF showed high anti-Stx2 titers and high bLF levels that were able to neutralize the cytotoxic effects of Stx2 in vitro and in vivo. Furthermore, HC-IgG/bLF avoided the inhibition of water absorption induced by E. coli O157:H7 in human colon and also the pathogenicity of E. coli O157:H7 and E. coli O157:H7Δstx2 in rat colon loops. Finally, HC-IgG/bLF prevented in a 100% the lethality caused by E. coli O157:H7 in a weaned mice model. Our study suggests that HC-IgG/bLF have protective effects against E. coli O157:H7 infection. These beneficial effects may be due to specific anti-Stx2 neutralizing antibodies in combination with high bLF levels. These results allow us to consider HC-IgG/bLF as a nutraceutical tool which could be used in combination with balanced supportive diets to prevent HUS. However further studies are required before recommendations can be made for therapeutic and clinical applications. Copyright © 2018 Elsevier Ltd. All rights reserved.

Effects of space flight, clinorotation, and centrifugation on the substrate utilization efficiency of E. coli

NASA Technical Reports Server (NTRS)

Brown, Robert B.; Klaus, D.; Todd, P.

2002-01-01

Cultures of Escherichia coli grown in space reached a 25% higher average final cell population than those in comparably matched ground controls (p<0.05). However, both groups consumed the same quantity of glucose, which suggests that space flight not only stimulated bacterial growth as has been previously reported, but also resulted in a 25% more efficient utilization of the available nutrients. Supporting experiments performed in "simulated weightlessness" under clinorotation produced similar trends of increased growth and efficiency, but to a lesser extent in absolute values. These experiments resulted in increases of 12% and 9% in average final cell population (p<0.05), while the efficiency of substrate utilization improved by 6% and 9% relative to static controls (p=0.12 and p<0.05, respectively). In contrast, hypergravity, produced by centrifugation, predictably resulted in the opposite effect--a decrease of 33% to 40% in final cell numbers with corresponding 29% to 40% lower net growth efficiencies (p<0.01). Collectively, these findings support the hypothesis that the increased bacterial growth observed in weightlessness is a result of reduced extracellular mass transport that occurs in the absence of sedimentation and buoyancy-driven convection, which consequently also improves substrate utilization efficiency in suspended cultures.

SAS molecular tests Escherichia coli O157 detection kit. Performance tested method 031203.

PubMed

Bapanpally, Chandra; Montier, Laura; Khan, Shah; Kasra, Akif; Brunelle, Sharon L

2014-01-01

The SAS Molecular tests Escherichia coli O157 Detection method, a loop-mediated isothermal amplification method, performed as well as or better than the U.S. Department of Agriculture, Food Safety Inspection Service Microbiology Laboratory Guidebook and the U.S. Food and Drug Administration Bacteriological Analytical Manual reference methods for ground beef, beef trim, bagged mixed lettuce, and fresh spinach. Ground beef (30% fat, 25 g test portion) was validated for 7-8 h enrichment, leafy greens were validated in a 6-7 h enrichment, and ground beef (30% fat, 375 g composite test portion) and beef trim (375 g composite test portion) were validated in a 16-20 h enrichment. The method performance for meat and leafy green matrixes was also shown to be acceptable under conditions of co-enrichment with Salmonella. Thus, after a short co-enrichment step, ground beef, beef trim, lettuce, and spinach can be tested for both Salmonella and E. coli O157. The SAS Molecular tests Salmonella Detection Kit was validated using the same test portions as for the SAS Molecular tests E. coli O157 Detection Kit and those results are presented in a separate report. Inclusivity and exclusivity testing revealed no false negatives and no false positives among the 50 E. coli 0157 strains, including H7 and non-motile strains, and 30 non-E. coli O157 strains examined. Finally, the method was shown to be robust when variations to DNA extract hold time and DNA volume were varied. The method comparison and robustness data suggest a full 7 h enrichment time should be used for 25 g ground beef test portions.

Can genetically modified Escherichia coli with neutral buoyancy induced by gas vesicles be used as an alternative method to clinorotation for microgravity studies?

PubMed

Benoit, Michael; Klaus, David

2005-01-01

Space flight has been shown to affect various bacterial growth parameters. It is proposed that weightlessness allows the cells to remain evenly distributed, consequently altering the chemical makeup of their surrounding fluid, and hence indirectly affecting their physiological behaviour. In support of this argument, ground-based studies using clinostats to partially simulate the quiescent environment attained in microgravity have generally been successful in producing bacterial growth characteristics that mimic responses reported under actual space conditions. A novel approach for evaluating the effects of reduced cell sedimentation is presented here through use of Escherichia coli cultures genetically modified to be neutrally buoyant. Since clinorotation would not (or would only minimally) affect cell distribution of this already near-colloidal cell system, it was hypothesized that the effects on final population density would be eliminated relative to a static control. Gas-vesicle-producing E. coli cultures were grown under clinostat and static conditions and the culture densities at 60 h were compared. As a control, E. coli that do not produce gas vesicles, but were otherwise identical to the experimental strain, were also grown under clinostat and static conditions. As hypothesized, no significant difference was observed in cell populations at 60 h between the clinorotated and static gas-vesicle-producing E. coli cultures, while the cells that did not produce gas vesicles showed a mean increase in population density of 10.5 % (P = 0.001). These results further suggest that the lack of cumulative cell sedimentation is the dominant effect of space flight on non-stirred, in vitro E. coli cultures.

RpoS Plays a Central Role in the SOS Induction by Sub-Lethal Aminoglycoside Concentrations in Vibrio cholerae

PubMed Central

Baharoglu, Zeynep; Krin, Evelyne; Mazel, Didier

2013-01-01

Bacteria encounter sub-inhibitory concentrations of antibiotics in various niches, where these low doses play a key role for antibiotic resistance selection. However, the physiological effects of these sub-lethal concentrations and their observed connection to the cellular mechanisms generating genetic diversification are still poorly understood. It is known that, unlike for the model bacterium Escherichia coli, sub-minimal inhibitory concentrations (sub-MIC) of aminoglycosides (AGs) induce the SOS response in Vibrio cholerae. SOS is induced upon DNA damage, and since AGs do not directly target DNA, we addressed two issues in this study: how sub-MIC AGs induce SOS in V. cholerae and why they do not do so in E. coli. We found that when bacteria are grown with tobramycin at a concentration 100-fold below the MIC, intracellular reactive oxygen species strongly increase in V. cholerae but not in E. coli. Using flow cytometry and gfp fusions with the SOS regulated promoter of intIA, we followed AG-dependent SOS induction. Testing the different mutation repair pathways, we found that over-expression of the base excision repair (BER) pathway protein MutY relieved this SOS induction in V. cholerae, suggesting a role for oxidized guanine in AG-mediated indirect DNA damage. As a corollary, we established that a BER pathway deficient E. coli strain induces SOS in response to sub-MIC AGs. We finally demonstrate that the RpoS general stress regulator prevents oxidative stress-mediated DNA damage formation in E. coli. We further show that AG-mediated SOS induction is conserved among the distantly related Gram negative pathogens Klebsiella pneumoniae and Photorhabdus luminescens, suggesting that E. coli is more of an exception than a paradigm for the physiological response to antibiotics sub-MIC. PMID:23613664

RpoS plays a central role in the SOS induction by sub-lethal aminoglycoside concentrations in Vibrio cholerae.

PubMed

Baharoglu, Zeynep; Krin, Evelyne; Mazel, Didier

2013-01-01

Bacteria encounter sub-inhibitory concentrations of antibiotics in various niches, where these low doses play a key role for antibiotic resistance selection. However, the physiological effects of these sub-lethal concentrations and their observed connection to the cellular mechanisms generating genetic diversification are still poorly understood. It is known that, unlike for the model bacterium Escherichia coli, sub-minimal inhibitory concentrations (sub-MIC) of aminoglycosides (AGs) induce the SOS response in Vibrio cholerae. SOS is induced upon DNA damage, and since AGs do not directly target DNA, we addressed two issues in this study: how sub-MIC AGs induce SOS in V. cholerae and why they do not do so in E. coli. We found that when bacteria are grown with tobramycin at a concentration 100-fold below the MIC, intracellular reactive oxygen species strongly increase in V. cholerae but not in E. coli. Using flow cytometry and gfp fusions with the SOS regulated promoter of intIA, we followed AG-dependent SOS induction. Testing the different mutation repair pathways, we found that over-expression of the base excision repair (BER) pathway protein MutY relieved this SOS induction in V. cholerae, suggesting a role for oxidized guanine in AG-mediated indirect DNA damage. As a corollary, we established that a BER pathway deficient E. coli strain induces SOS in response to sub-MIC AGs. We finally demonstrate that the RpoS general stress regulator prevents oxidative stress-mediated DNA damage formation in E. coli. We further show that AG-mediated SOS induction is conserved among the distantly related Gram negative pathogens Klebsiella pneumoniae and Photorhabdus luminescens, suggesting that E. coli is more of an exception than a paradigm for the physiological response to antibiotics sub-MIC.

Rapid pathogen detection with bacterial-assembled magnetic mesoporous silica.

PubMed

Lee, Soo Youn; Lee, Jiho; Lee, Hye Sun; Chang, Jeong Ho

2014-03-15

We report rapid and accurate pathogen detection by coupling with high efficiency magnetic separation of pathogen by Ni(2+)-heterogeneous magnetic mesoporous silica (Ni-HMMS) and real time-polymerase chain reaction (RT-PCR) technique. Ni-HMMS was developed with a significant incorporation of Fe particles within the silica mesopores by programmed thermal hydrogen reaction and functionalized with Ni(2+) ion on the surface by the wet impregnation process. High abundant Ni(2+) ions on the Ni-HMMS surface were able to assemble with cell wall component protein NikA (nickel-binding membrane protein), which contains several pathogenic bacteria including Escherichia coli O157:H7. NikA protein expression experiment showed the outstanding separation rate of the nikA gene-overexpressed E. coli (pSY-Nik) when comparing with wild-type E. coli (44.5 ± 13%) or not over-expressed E. coli (pSY-Nik) (53.2 ± 2.7%). Moreover, Ni-HMMS showed lower obstacle effect by large reaction volume (10 mL) than spherical core/shell-type silica magnetic nanoparticles functionalized with Ni(2+) (ca. 40 nm-diameters). Finally, the Ni-HMMS was successfully assessed to separate pathogenic E. coli O157:H7 and applied to direct and rapid RT-PCR to quantitative detection at ultralow concentration (1 Log10 cfu mL(-1)) in the real samples (milk and Staphylococcus aureus culture broth) without bacterial amplification and DNA extraction step. © 2013 Elsevier B.V. All rights reserved.

Engineering Escherichia coli Co-Cultures for Production of Curcuminoids From Glucose.

PubMed

Fang, Zhen; Jones, John A; Zhou, Jingwen; Koffas, Mattheos A G

2018-05-01

Curcuminoids (cus) have attracted increasing attention because of the antioxidant, anticancer, and antitumor activities while their production is limited because of its main source, turmeric plant, demonstrates extensive seasonal variation. In this study, we constructed Escherichia coli co-culture system for the rapid production of curcuminoids from glucose. Firstly, the overexpression of curcuminoid synthase and four different strategies related to increasing the intracellular malonyl-CoA pool were conducted in engineered E. coli. We found that bisdemethoxycurcumin (BDMC) is the main product and that high level of malonyl-CoA pool is essential for BDMC production. We also obtained the maximum titer (13.8 mg L -1 ) of BDMC within 4 h by fast preparation directly from p-coumaric acid. Secondly, we developed a process for BDMC synthesis from glucose using a co-culture system where an E. coli strain is used to produce p-coumaric acid from glucose and another E. coli strain converted p-coumaric acid into the final product. Compared to the mono-culture system, the co-culture is more potent and resulted in 6.28 mg L -1 of BDMC from glucose within 22 h of fermentation in a 3-L bioreactor. This is the first time a co-culture method is employed for the production of curcuminoids from glucose in a lab scale bioreactor. This system provides a new method transforming inexpensive substrate into value-added products. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Ultra-low background DNA cloning system.

PubMed

Goto, Kenta; Nagano, Yukio

2013-01-01

Yeast-based in vivo cloning is useful for cloning DNA fragments into plasmid vectors and is based on the ability of yeast to recombine the DNA fragments by homologous recombination. Although this method is efficient, it produces some by-products. We have developed an "ultra-low background DNA cloning system" on the basis of yeast-based in vivo cloning, by almost completely eliminating the generation of by-products and applying the method to commonly used Escherichia coli vectors, particularly those lacking yeast replication origins and carrying an ampicillin resistance gene (Amp(r)). First, we constructed a conversion cassette containing the DNA sequences in the following order: an Amp(r) 5' UTR (untranslated region) and coding region, an autonomous replication sequence and a centromere sequence from yeast, a TRP1 yeast selectable marker, and an Amp(r) 3' UTR. This cassette allowed conversion of the Amp(r)-containing vector into the yeast/E. coli shuttle vector through use of the Amp(r) sequence by homologous recombination. Furthermore, simultaneous transformation of the desired DNA fragment into yeast allowed cloning of this DNA fragment into the same vector. We rescued the plasmid vectors from all yeast transformants, and by-products containing the E. coli replication origin disappeared. Next, the rescued vectors were transformed into E. coli and the by-products containing the yeast replication origin disappeared. Thus, our method used yeast- and E. coli-specific "origins of replication" to eliminate the generation of by-products. Finally, we successfully cloned the DNA fragment into the vector with almost 100% efficiency.

Common and divergent features in transcriptional control of the homologous small RNAs GlmY and GlmZ in Enterobacteriaceae

PubMed Central

Göpel, Yvonne; Lüttmann, Denise; Heroven, Ann Kathrin; Reichenbach, Birte; Dersch, Petra; Görke, Boris

2011-01-01

Small RNAs GlmY and GlmZ compose a cascade that feedback-regulates synthesis of enzyme GlmS in Enterobacteriaceae. Here, we analyzed the transcriptional regulation of glmY/glmZ from Yersinia pseudotuberculosis, Salmonella typhimurium and Escherichia coli, as representatives for other enterobacterial species, which exhibit similar promoter architectures. The GlmY and GlmZ sRNAs of Y. pseudotuberculosis are transcribed from σ54-promoters that require activation by the response regulator GlrR through binding to three conserved sites located upstream of the promoters. This also applies to glmY/glmZ of S. typhimurium and glmY of E. coli, but as a difference additional σ70-promoters overlap the σ54-promoters and initiate transcription at the same site. In contrast, E. coli glmZ is transcribed from a single σ70-promoter. Thus, transcription of glmY and glmZ is controlled by σ54 and the two-component system GlrR/GlrK (QseF/QseE) in Y. pseudotuberculosis and presumably in many other Enterobacteria. However, in a subset of species such as E. coli this relationship is partially lost in favor of σ70-dependent transcription. In addition, we show that activity of the σ54-promoter of E. coli glmY requires binding of the integration host factor to sites upstream of the promoter. Finally, evidence is provided that phosphorylation of GlrR increases its activity and thereby sRNA expression. PMID:20965974

A Pilot Program to Provide Ownership Skills to Seniors in the Vocational Technical Schools and Other Educational Institutions, Year Two. Final Report.

ERIC Educational Resources Information Center

Norwich Regional Vocational Technical School, CT.

Responding to a Small Business Administration statement that technical school graduates lack knowledge of business principles, the Norwich (Connecticut) Regional Vocational Technical School conducted a course in small business ownership/management for all of its seniors. The required course, a fifty-four-hour module, replaced one English,…

A Case Study of Technical Assistance to Demonstration Programs for Young Handicapped Children. Part III.

ERIC Educational Resources Information Center

Behr, Shirley K.; And Others

The report describes the third and final year of a 3-year case study of the technical assistance process as implemented by the Technical Assistance Development System (TADS) for the staffs of two demonstration programs for preschool handicapped children and their families. Following a review of TADS and the two demonstration programs, the…

Chemical Durability Improvement and Static Fatigue of Glasses.

DTIC Science & Technology

1982-08-01

Afl-Alla 837 RENSSELAER POLYIECmfJ!C INST TRtOY NY DEPT OF MATERIAL--ETC F/6 ii/ CHEMICAL DURABILITY IMPROVEMENT AND STATIC FATIGUE OF GLASSESW AUC2...82 M TOMOZAWA NOGGIN 7A-C-0315 UNC LASS IF IED N ENEEEEEE FINAL TECHNICAL REPORT For the period April 1, 1978 "u March 31, 198200 CHEMICAL DURABILITY...REPORT A PERIOD COVERED Chemical Durability Improvement and Static Final Technical Report Fatiue o GlasesApril 1, 1978"’,March 31, 1982 S. PERFORMING ORG

Experimental Program Final Technical Progress Report: 15 February 2007 to 30 September 2012

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kinney, Edward R.

2014-09-12

This is the final technical report of the grant DE-FG02-04ER41301 to the University of Colorado at Boulder entitled "Intermediate Energy Nuclear Physics" and describes the results of our funded activities during the period 15 February 2007 to 30 September 2012. These activities were primarily carried out at Fermilab, RHIC, and the German lab DESY. Significant advances in these experiments were carried out by members of the Colorado group and are described in detail.

Final Technical Report - Center for Technology for Advanced Scientific Component Software (TASCS)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sussman, Alan

2014-10-21

This is a final technical report for the University of Maryland work in the SciDAC Center for Technology for Advanced Scientific Component Software (TASCS). The Maryland work focused on software tools for coupling parallel software components built using the Common Component Architecture (CCA) APIs. Those tools are based on the Maryland InterComm software framework that has been used in multiple computational science applications to build large-scale simulations of complex physical systems that employ multiple separately developed codes.

Iowa Hill Pumped Storage Project Investigations - Final Technical Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hanson, David

2016-07-01

This Final Technical Report is a summary of the activities and outcome of the Department of Energy (DOE) Assistance Agreement DE-EE0005414 with the Sacramento Municipal Utility District (SMUD). The Assistance Agreement was created in 2012 to support investigations into the Iowa Hill Pumped-storage Project (Project), a new development that would add an additional 400 MW of capacity to SMUD’s existing 688MW Upper American River Hydroelectric Project (UARP) in the Sierra Nevada mountains east of Sacramento, California.

Shiga toxin-producing Escherichia coli isolated from chicken meat in Iran: serogroups, virulence factors, and antimicrobial resistance properties.

PubMed

Momtaz, Hassan; Jamshidi, Alireza

2013-05-01

The aim of the current study was to determine the virulence factors, serogroups, and antibiotic resistance properties of Shiga toxin-producing Escherichia coli isolated from chicken meat samples. A total of 422 chicken meat samples were collected from 5 townships of Iran. Specimens were immediately transferred to the laboratory in a cooler with an ice pack. Samples were cultured, and the positive culture samples were analyzed by PCR assays. Finally, the antimicrobial susceptibility test was performed using the disk diffusion method in Mueller-Hinton agar. According to the results, out of 422 samples, 146 (34.59%) were confirmed to be E. coli positive and among E. coli-positive samples, 51 (34.93%) and 31 (21.23%) were from attaching and effacing E. coli (AEEC) and enterohemorrhagic E. coli (EHEC) subgroups, respectively. All of the EHEC-positive samples had all stx1, eaeA, and ehly virulence genes, whereas only 5 (9.80%) of AEEC subgroup had all stx1, stx2, and eaeA genes. As the data revealed, O157 was the most prevalent and O111 was the least prevalent strains in the Shiga toxin-producing E. coli (STEC) population. Among STEC strains, sulI and blaSHV had the highest and lowest incidence rate, respectively. There was a high resistance to tetracycline (76.82%), followed by chloramphenicol (73.17%) and nitrofurantoin (63.41%), but there was low resistance to cephalotine (7.31%) antibiotics in isolated strains. Results shows that the PCR technique has a high performance for detection of serogroups, virulence genes, and antibiotic resistance genes in STEC strains. This study is the first prevalence report of detection of virulence genes, serogroups, and antibiotic resistance properties of STEC strains isolated from chicken meat samples in Iran. Based on the results, chicken meat is one of the main sources of STEC strains and its virulence factors in Iran, so an accurate meat inspection would reduce disease outbreaks.

Renewable Fuel Standard Program (RFS1): Final Rule Additional Resources

EPA Pesticide Factsheets

The final rule of fuels and fuel additives: renewable fuel standard program is published on May 1, 2007 and is effective on September 1, 2007. You will find the links to this final rule and technical amendments supporting this rule.

Assurance Penalty Level Analysis Final Rule TSD

EPA Pesticide Factsheets

This Technical Support Document (TSD) supports EPA’s determination that the final Transport Rule’s assurance provision penalty requirement provides sufficient deterrence against a state exceeding its assurance level.

Effects of space flight and mixing on bacterial growth in low volume cultures

NASA Technical Reports Server (NTRS)

Kacena, M. A.; Manfredi, B.; Todd, P.

1999-01-01

Previous investigations have shown that liquid suspension bacterial cultures grow to higher cell concentrations in spaceflight than on Earth. None of these studies included ground-control experiments designed to evaluate the fluid effects potentially responsible for the reported increases. Therefore, the emphasis of this research was to both confirm differences in final cell concentration between 1g and microgravity cultures, and to examine the effects of mixing as a partial explanation for this difference. Flight experiments were performed in the Fluid Processing Apparatus (FPA), aboard Space Shuttle Missions STS-63 and STS-69, with simultaneous 1g static and agitated controls. Additional static 1g, agitated, and clino-rotated controls were performed in 9-ml culture tubes. This research revealed that both E. coli and B. subtilis samples cultured in space flight grew to higher final cell densities (120-345% increase) than simultaneous static 1g controls. The final cell concentration of E. coli cells cultured under agitation was 43% higher than in static 1g cultures and was 102% higher with clino-rotation. However, for B. subtilis cultures grown while being agitated on a shaker or clino-rotated, the final cell concentrations were nearly identical to those of the simultaneous static 1g controls. Therefore, these data suggest that the unique fluid quiescence in the microgravity environment (lack of sedimentation, creating unique transfer of nutrients and waste products), was responsible for the enhanced bacterial proliferation reported in this and other studies.

Final Rule for Technical Amendments to the Highway and Nonroad Diesel Regulations

EPA Pesticide Factsheets

This action corrects errors and omissions from the previous rules, makes minor changes to the regulations to assist entities with regulatory compliance, and makes technical amendments that resulted from discussions with various diesel stakeholders.

Survival, Biofilm Formation, and Growth Potential of Environmental and Enteric Escherichia coli Strains in Drinking Water Microcosms

PubMed Central

Abberton, Cathy L.; Bereschenko, Ludmila; van der Wielen, Paul W. J. J.

2016-01-01

ABSTRACT Escherichia coli is the most commonly used indicator for fecal contamination in drinking water distribution systems (WDS). The assumption is that E. coli bacteria are of enteric origin and cannot persist for long outside their host and therefore act as indicators of recent contamination events. This study investigates the fate of E. coli in drinking water, specifically addressing survival, biofilm formation under shear stress, and regrowth in a series of laboratory-controlled experiments. We show the extended persistence of three E. coli strains (two enteric isolates and one soil isolate) in sterile and nonsterile drinking water microcosms at 8 and 17°C, with T90 (time taken for a reduction in cell number of 1 log10 unit) values ranging from 17.4 ± 1.8 to 149 ± 67.7 days, using standard plate counts and a series of (reverse transcription-)quantitative PCR [(RT-)Q-PCR] assays targeting 16S rRNA, tuf, uidA, and rodA genes and transcripts. Furthermore, each strain was capable of attaching to a surface and replicating to form biofilm in the presence of nutrients under a range of shear stress values (0.6, 2.0, and 4.4 dynes [dyn] cm−2; BioFlux system; Fluxion); however, cell numbers did not increase when drinking water flowed over the biofilm (P > 0.05 by t test). Finally, E. coli regrowth within drinking water microcosms containing polyethylene PE-100 pipe wall material was not observed in the biofilm or water phase using a combination of culturing and Q-PCR methods for E. coli. The results of this work highlight that when E. coli enters drinking water it has the potential to survive and attach to surfaces but that regrowth within drinking water or biofilm is unlikely. IMPORTANCE The provision of clean, safe drinking water is fundamental to society. WDS deliver water to consumers via a vast network of pipes. E. coli is used as an indicator organism for recent contamination events based on the premise that it cannot survive for long outside its host. A key public health concern therefore arises around the fate of E. coli on entering a WDS; its survival, ability to form a biofilm, and potential for regrowth. In particular, if E. coli bacteria have the ability to incorporate and regrow within the pipe wall biofilm of a WDS, they could reinoculate the water at a later stage. This study sheds light on the fate of environmental and enteric strains of E. coli in drinking water showing extended survival, the potential for biofilm formation under shear stress, and importantly, that regrowth in the presence of an indigenous microbial community is unlikely. PMID:27342552

Survival, Biofilm Formation, and Growth Potential of Environmental and Enteric Escherichia coli Strains in Drinking Water Microcosms.

PubMed

Abberton, Cathy L; Bereschenko, Ludmila; van der Wielen, Paul W J J; Smith, Cindy J

2016-09-01

Escherichia coli is the most commonly used indicator for fecal contamination in drinking water distribution systems (WDS). The assumption is that E. coli bacteria are of enteric origin and cannot persist for long outside their host and therefore act as indicators of recent contamination events. This study investigates the fate of E. coli in drinking water, specifically addressing survival, biofilm formation under shear stress, and regrowth in a series of laboratory-controlled experiments. We show the extended persistence of three E. coli strains (two enteric isolates and one soil isolate) in sterile and nonsterile drinking water microcosms at 8 and 17°C, with T90 (time taken for a reduction in cell number of 1 log10 unit) values ranging from 17.4 ± 1.8 to 149 ± 67.7 days, using standard plate counts and a series of (reverse transcription-)quantitative PCR [(RT-)Q-PCR] assays targeting 16S rRNA, tuf, uidA, and rodA genes and transcripts. Furthermore, each strain was capable of attaching to a surface and replicating to form biofilm in the presence of nutrients under a range of shear stress values (0.6, 2.0, and 4.4 dynes [dyn] cm(-2); BioFlux system; Fluxion); however, cell numbers did not increase when drinking water flowed over the biofilm (P > 0.05 by t test). Finally, E. coli regrowth within drinking water microcosms containing polyethylene PE-100 pipe wall material was not observed in the biofilm or water phase using a combination of culturing and Q-PCR methods for E. coli The results of this work highlight that when E. coli enters drinking water it has the potential to survive and attach to surfaces but that regrowth within drinking water or biofilm is unlikely. The provision of clean, safe drinking water is fundamental to society. WDS deliver water to consumers via a vast network of pipes. E. coli is used as an indicator organism for recent contamination events based on the premise that it cannot survive for long outside its host. A key public health concern therefore arises around the fate of E. coli on entering a WDS; its survival, ability to form a biofilm, and potential for regrowth. In particular, if E. coli bacteria have the ability to incorporate and regrow within the pipe wall biofilm of a WDS, they could reinoculate the water at a later stage. This study sheds light on the fate of environmental and enteric strains of E. coli in drinking water showing extended survival, the potential for biofilm formation under shear stress, and importantly, that regrowth in the presence of an indigenous microbial community is unlikely. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Novel method using nanomaterials for removal of E. coli bacteria from water

NASA Astrophysics Data System (ADS)

Al-Hakami, Samer M.

A novel technology has been presented for the removal of Escherichia Coli (E. coli) bacteria from water using Carbon Nanotubes (CNTs), modified CNTs and impregnated CNTs with and without heating effect from microwave radiation. The presented microbiological treatment in this study, which lies under the subject field of bio-nanotechnology has combined between powers of degradation by microwaves and adsorption by Carbon Nanotubes (CNTs). The study demonstrated the removal of Escherichia Coli (E. coli) Bacteria and its by-products under various treatment conditions including dosages of CNTs, the effect of functional groups, the effect of the power of microwave source, and treatment time, the final results indicated that different types of the tested nanomaterials (raw CNTs, functionalized CNTs and impregnated CNTs with and without heating effect from microwave radiation with a very small dosage between 0.2 g and 0.007 g of CNTs per 100 ml of water) successfully achieved 90-100% removal of E. coli bacteria from water. This significant result can be utilized in many of medical and water treatment applications, such as: disinfection for virus, microbial control, water purification and treatment several types of cancer as part of tumor therapy. The study was carried out in four phases. The first phase focused on the synthesis and preparation of the functional groups and the impregnation of nano particles (CNTs and Ag). In this phase, the multi-walled carbon nanotubes were functionalized with a carboxylic (COOH) group, phenol (C5H 5OH) group, dodecylamine group (C12H27N), C18 group such as I-octadecanol (C18H38O), and impregnated with silver nanoparticles. The second phase included the laboratory bench-scale experimental work where several samples under the effect of microwaves were tested under various conditions. In this phase, E. coli bacteria numbers evaluated before and after treatment under the individual and combined effects of treatment schemes. The third phase included characterization of nanomatgerials and bacterial colonies. The fourth phase included the data analysis in order to identify the treatment conditions that led to complete removal of E. coli bacteria from water.

Techno-economic analysis of the industrial production of a low-cost enzyme using E. coli: the case of recombinant β-glucosidase.

PubMed

Ferreira, Rafael da Gama; Azzoni, Adriano Rodrigues; Freitas, Sindelia

2018-01-01

The enzymatic conversion of lignocellulosic biomass into fermentable sugars is a promising approach for producing renewable fuels and chemicals. However, the cost and efficiency of the fungal enzyme cocktails that are normally employed in these processes remain a significant bottleneck. A potential route to increase hydrolysis yields and thereby reduce the hydrolysis costs would be to supplement the fungal enzymes with their lacking enzymatic activities, such as β-glucosidase. In this context, it is not clear from the literature whether recombinant E. coli could be a cost-effective platform for the production of some of these low-value enzymes, especially in the case of on-site production. Here, we present a conceptual design and techno-economic evaluation of the production of a low-cost industrial enzyme using recombinant E. coli . In a simulated baseline scenario for β-glucosidase demand in a hypothetical second-generation ethanol (2G) plant in Brazil, we found that the production cost (316 US$/kg) was higher than what is commonly assumed in the literature for fungal enzymes, owing especially to the facility-dependent costs (45%) and to consumables (23%) and raw materials (25%). Sensitivity analyses of process scale, inoculation volume, and volumetric productivity indicated that optimized conditions may promote a dramatic reduction in enzyme cost and also revealed the most relevant factors affecting production costs. Despite the considerable technical and economic uncertainties that surround 2G ethanol and the large-scale production of low-cost recombinant enzymes, this work sheds light on some relevant questions and supports future studies in this field. In particular, we conclude that process optimization, on many fronts, may strongly reduce the costs of E. coli recombinant enzymes, in the context of tailor-made enzymatic cocktails for 2G ethanol production.

Final priority. Rehabilitation Training: Job-Driven Vocational Rehabilitation Technical Assistance Center. Final priority.

PubMed

2014-08-19

The Assistant Secretary for Special Education and Rehabilitative Services announces a priority under the Rehabilitation Training program to establish a Job-Driven Vocational Rehabilitation Technical Assistance Center (JDVRTAC). The Assistant Secretary may use this priority for competitions in fiscal year (FY) 2014 and later years. We take this action to focus on training in an area of national need. Specifically, this priority responds to the Presidential Memorandum to Federal agencies directing them to take action to address job-driven training for the Nation's workers. The JDVRTAC will provide technical assistance (TA) to State vocational rehabilitation (VR) agencies to help them develop for individuals with disabilities training and employment opportunities that meet the needs of today's employers.

The utility of multiple molecular methods including whole genome sequencing as tools to differentiate Escherichia coli O157:H7 outbreaks.

PubMed

Berenger, Byron M; Berry, Chrystal; Peterson, Trevor; Fach, Patrick; Delannoy, Sabine; Li, Vincent; Tschetter, Lorelee; Nadon, Celine; Honish, Lance; Louie, Marie; Chui, Linda

2015-01-01

A standardised method for determining Escherichia coli O157:H7 strain relatedness using whole genome sequencing or virulence gene profiling is not yet established. We sought to assess the capacity of either high-throughput polymerase chain reaction (PCR) of 49 virulence genes, core-genome single nt variants (SNVs) or k-mer clustering to discriminate between outbreak-associated and sporadic E. coli O157:H7 isolates. Three outbreaks and multiple sporadic isolates from the province of Alberta, Canada were included in the study. Two of the outbreaks occurred concurrently in 2014 and one occurred in 2012. Pulsed-field gel electrophoresis (PFGE) and multilocus variable-number tandem repeat analysis (MLVA) were employed as comparator typing methods. The virulence gene profiles of isolates from the 2012 and 2014 Alberta outbreak events and contemporary sporadic isolates were mostly identical; therefore the set of virulence genes chosen in this study were not discriminatory enough to distinguish between outbreak clusters. Concordant with PFGE and MLVA results, core genome SNV and k-mer phylogenies clustered isolates from the 2012 and 2014 outbreaks as distinct events. k-mer phylogenies demonstrated increased discriminatory power compared with core SNV phylogenies. Prior to the widespread implementation of whole genome sequencing for routine public health use, issues surrounding cost, technical expertise, software standardisation, and data sharing/comparisons must be addressed.

Metabolic Engineering of Escherichia coli K12 for Homofermentative Production of L-Lactate from Xylose.

PubMed

Jiang, Ting; Zhang, Chen; He, Qin; Zheng, Zhaojuan; Ouyang, Jia

2018-02-01

The efficient utilization of xylose is regarded as a technical barrier to the commercial production of bulk chemicals from biomass. Due to the desirable mechanical properties of polylactic acid (PLA) depending on the isomeric composition of lactate, biotechnological production of lactate with high optical pure has been increasingly focused in recent years. The main objective of this work was to construct an engineered Escherichia coli for the optically pure L-lactate production from xylose. Six chromosomal deletions (pflB, ldhA, ackA, pta, frdA, adhE) and a chromosomal integration of L-lactate dehydrogenase-encoding gene (ldhL) from Bacillus coagulans was involved in construction of E. coli KSJ316. The recombinant strain could produce L-lactate from xylose resulting in a yield of 0.91 g/g xylose. The chemical purity of L-lactate was 95.52%, and the optical purity was greater than 99%. Moreover, three strategies, including overexpression of L-lactate dehydrogenase, intensification of xylose catabolism, and addition of additives to medium, were designed to enhance the production. The results showed that they could increase the concentration of L-lactate by 32.90, 20.13, and 233.88% relative to the control, respectively. This was the first report that adding formate not only could increase the xylose utilization but also led to the fewer by-product levels.

The functional landscape bound to the transcription factors of Escherichia coli K-12.

PubMed

Pérez-Rueda, Ernesto; Tenorio-Salgado, Silvia; Huerta-Saquero, Alejandro; Balderas-Martínez, Yalbi I; Moreno-Hagelsieb, Gabriel

2015-10-01

Motivated by the experimental evidences accumulated in the last ten years and based on information deposited in RegulonDB, literature look up, and sequence analysis, we analyze the repertoire of 304 DNA-binding Transcription factors (TFs) in Escherichia coli K-12. These regulators were grouped in 78 evolutionary families and are regulating almost half of the total genes in this bacterium. In structural terms, 60% of TFs are composed by two-domains, 30% are monodomain, and 10% three- and four-structural domains. As previously noticed, the most abundant DNA-binding domain corresponds to the winged helix-turn-helix, with few alternative DNA-binding structures, resembling the hypothesis of successful protein structures with the emergence of new ones at low scales. In summary, we identified and described the characteristics associated to the DNA-binding TF in E. coli K-12. We also identified twelve functional modules based on a co-regulated gene matrix. Finally, diverse regulons were predicted based on direct associations between the TFs and potential regulated genes. This analysis should increase our knowledge about the gene regulation in the bacterium E. coli K-12, and provide more additional clues for comprehensive modelling of transcriptional regulatory networks in other bacteria. Copyright © 2015 Elsevier Ltd. All rights reserved.

Integrated analysis of isopentenyl pyrophosphate (IPP) toxicity in isoprenoid-producing Escherichia coli

DOE Office of Scientific and Technical Information (OSTI.GOV)

George, Kevin W.; Thompson, Mitchell; Kim, Joonhoon

Isopentenyl pyrophosphate (IPP) toxicity presents a challenge in engineered microbial systems since its formation is unavoidable in terpene biosynthesis. Here in this work, we develop an experimental platform to study IPP toxicity in isoprenol-producing Escherichia coli. We first characterize the physiological response to IPP accumulation, demonstrating that elevated IPP levels are linked to growth inhibition, reduced cell viability, and plasmid instability. We show that IPP toxicity selects for pathway “breakage”, using proteomics to identify a reduction in phosphomevalonate kinase (PMK) as a probable recovery mechanism. Next, using multi-omics data, we demonstrate that endogenous E. coli metabolism is globally impacted bymore » IPP accumulation, which slows nutrient uptake, decreases ATP levels, and perturbs nucleotide metabolism. We also observe the extracellular accumulation of IPP and present preliminary evidence that IPP can be transported by E. coli, findings that might be broadly relevant for the study of isoprenoid biosynthesis. Finally, we discover that IPP accumulation leads to the formation of ApppI, a nucleotide analog of IPP that may contribute to observed toxicity phenotypes. This comprehensive assessment of IPP stress suggests potential strategies for the alleviation of prenyl diphosphate toxicity and highlights possible engineering targets for improved IPP flux and high titer isoprenoid production.« less

Integrated analysis of isopentenyl pyrophosphate (IPP) toxicity in isoprenoid-producing Escherichia coli

DOE PAGES

George, Kevin W.; Thompson, Mitchell; Kim, Joonhoon; ...

2018-03-09

Isopentenyl pyrophosphate (IPP) toxicity presents a challenge in engineered microbial systems since its formation is unavoidable in terpene biosynthesis. Here in this work, we develop an experimental platform to study IPP toxicity in isoprenol-producing Escherichia coli. We first characterize the physiological response to IPP accumulation, demonstrating that elevated IPP levels are linked to growth inhibition, reduced cell viability, and plasmid instability. We show that IPP toxicity selects for pathway “breakage”, using proteomics to identify a reduction in phosphomevalonate kinase (PMK) as a probable recovery mechanism. Next, using multi-omics data, we demonstrate that endogenous E. coli metabolism is globally impacted bymore » IPP accumulation, which slows nutrient uptake, decreases ATP levels, and perturbs nucleotide metabolism. We also observe the extracellular accumulation of IPP and present preliminary evidence that IPP can be transported by E. coli, findings that might be broadly relevant for the study of isoprenoid biosynthesis. Finally, we discover that IPP accumulation leads to the formation of ApppI, a nucleotide analog of IPP that may contribute to observed toxicity phenotypes. This comprehensive assessment of IPP stress suggests potential strategies for the alleviation of prenyl diphosphate toxicity and highlights possible engineering targets for improved IPP flux and high titer isoprenoid production.« less

A Study on the Interaction of Rhodamine B with Methylthioadenosine Phosphorylase Protein Sourced from an Antarctic Soil Metagenomic Library

PubMed Central

Bujacz, Anna; Wierzbicka-Woś, Anna; Kur, Józef

2013-01-01

The presented study examines the phenomenon of the fluorescence under UV light excitation (312 nm) of E. coli cells expressing a novel metagenomic-derived putative methylthioadenosine phosphorylase gene, called rsfp, grown on LB agar supplemented with a fluorescent dye rhodamine B. For this purpose, an rsfp gene was cloned and expressed in an LMG194 E. coli strain using an arabinose promoter. The resulting RSFP protein was purified and its UV-VIS absorbance spectrum and emission spectrum were assayed. Simultaneously, the same spectroscopic studies were carried out for rhodamine B in the absence or presence of RSFP protein or native E. coli proteins, respectively. The results of the spectroscopic studies suggested that the fluorescence of E. coli cells expressing rsfp gene under UV illumination is due to the interaction of rhodamine B molecules with the RSFP protein. Finally, this interaction was proved by a crystallographic study and then by site-directed mutagenesis of rsfp gene sequence. The crystal structures of RSFP apo form (1.98 Å) and complex RSFP/RB (1.90 Å) show a trimer of RSFP molecules located on the crystallographic six fold screw axis. The RSFP complex with rhodamine B revealed the binding site for RB, in the pocket located on the interface between symmetry related monomers. PMID:23383268

Solar water disinfection (SODIS) of Escherichia coli, Enterococcus spp., and MS2 coliphage: effects of additives and alternative container materials.

PubMed

Fisher, Michael B; Iriarte, Mercedes; Nelson, Kara L

2012-04-15

The use of alternative container materials and added oxidants accelerated the inactivation of MS2 coliphage and Escherichia coli and Enterococcus spp. bacteria during solar water disinfection (SODIS) trials. Specifically, bottles made from polypropylene copolymer (PPCO), a partially UVB-transparent plastic, resulted in three-log inactivation of these organisms in approximately half the time required for disinfection in bottles made from PET, polycarbonate, or Tritan(®), which absorb most UVB light. Furthermore, the addition of 125 mg/L sodium percarbonate in combination with either citric acid or copper plus ascorbate tended to accelerate inactivation by factors of 1.4-19. Finally, it was observed that the inactivation of E. coli and enterococci derived from local wastewater was far slower than the inactivation of laboratory-cultured E. coli and Enterococcus spp., while the inactivation of MS2 was slowest of all. These results highlight the importance of UVB in SODIS under certain conditions, and also the greater sunlight resistance of some viruses and of bacteria of fecal origin, as compared to the laboratory-cultured bacteria commonly used to model their inactivation. Furthermore, this study illustrates promising new avenues for accelerating the inactivation of bacteria and viruses by solar disinfection. Copyright © 2011 Elsevier Ltd. All rights reserved.

Metabolic engineering of Escherichia coli to produce 2'-fucosyllactose via salvage pathway of guanosine 5'-diphosphate (GDP)-l-fucose.

PubMed

Chin, Young-Wook; Seo, Nari; Kim, Jae-Han; Seo, Jin-Ho

2016-11-01

2'-Fucosyllactose (2-FL) is one of the key oligosaccharides in human milk. In the present study, the salvage guanosine 5'-diphosphate (GDP)-l-fucose biosynthetic pathway from fucose was employed in engineered Escherichia coli BL21star(DE3) for efficient production of 2-FL. Introduction of the fkp gene coding for fucokinase/GDP-l-fucose pyrophosphorylase (Fkp) from Bacteroides fragilis and the fucT2 gene encoding α-1,2-fucosyltransferase from Helicobacter pylori allows the engineered E. coli to produce 2-FL from fucose, lactose and glycerol. To enhance the lactose flux to 2-FL production, the attenuated, and deleted mutants of β-galactosidase were employed. Moreover, the 2-FL yield and productivity were further improved by deletion of the fucI-fucK gene cluster coding for fucose isomerase (FucI) and fuculose kinase (FucK). Finally, fed-batch fermentation of engineered E. coli BL21star(DE3) deleting lacZ and fucI-fucK, and expressing fkp and fucT2 resulted in 23.1 g/L of extracellular concentration of 2-FL and 0.39 g/L/h productivity. Biotechnol. Bioeng. 2016;113: 2443-2452. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Metabolic engineering of the L-phenylalanine pathway in Escherichia coli for the production of S- or R-mandelic acid

PubMed Central

2011-01-01

Background Mandelic acid (MA), an important component in pharmaceutical syntheses, is currently produced exclusively via petrochemical processes. Growing concerns over the environment and fossil energy costs have inspired a quest to develop alternative routes to MA using renewable resources. Herein we report the first direct route to optically pure MA from glucose via genetic modification of the L-phenylalanine pathway in E. coli. Results The introduction of hydroxymandelate synthase (HmaS) from Amycolatopsis orientalis into E. coli led to a yield of 0.092 g/L S-MA. By combined deletion of competing pathways, further optimization of S-MA production was achieved, and the yield reached 0.74 g/L within 24 h. To produce R-MA, hydroxymandelate oxidase (Hmo) from Streptomyces coelicolor and D-mandelate dehydrogenase (DMD) from Rhodotorula graminis were co-expressed in an S-MA-producing strain, and the resulting strain was capable of producing 0.68 g/L R-MA. Finally, phenylpyruvate feeding experiments suggest that HmaS is a potential bottleneck to further improvement in yields. Conclusions We have constructed E. coli strains that successfully accomplished the production of S- and R-MA directly from glucose. Our work provides the first example of the completely fermentative production of S- and R-MA from renewable feedstock. PMID:21910908

NASA's Pursuit of Low-Noise Propulsion for Low-Boom Commercial Supersonic Vehicles

NASA Technical Reports Server (NTRS)

Bridges, James; Brown, Clifford A.; Seidel, Jonathan A.

2018-01-01

Since 2006, when the Fundamental Aeronautics Program was instituted within NASA's Aeronautics Mission Directorate, there has been a Project looking at the technical barriers to commercial supersonic flight. Among the barriers is the noise produced by aircraft during landing and takeoff. Over the years that followed, research was carried out at NASA aeronautics research centers, often in collaboration with academia and industry, addressing the problem. In 2013, a high-level milestone was established, described as a Technical Challenge, with the objective of demonstrating the feasibility of a low-boom supersonic airliner that could meet current airport noise regulations. The Technical Challenge was formally called "Low Noise Propulsion for Low Boom Aircraft", and was completed in late 2016. This paper reports the technical findings from this Technical Challenge, reaching back almost 10 years to review the technologies and tools that were developed along the way. It also discusses the final aircraft configuration and propulsion systems required for a supersonic civilian aircraft to meet noise regulations using the technologies available today. Finally, the paper documents the model-scale tests that validated the acoustic performance of the study aircraft.

NASA's Pursuit of Low-Noise Propulsion for Low-Boom Commercial Supersonic Vehicles

NASA Technical Reports Server (NTRS)

Bridges, James; Brown, Clifford A.; Seidel, Jonathan

2018-01-01

Since 2006, when the Fundamental Aeronautics Program was instituted within NASA's Aeronautics Mission Directorate, there has been a Project looking at the technical barriers to commercial supersonic flight. Among the barriers is the noise produced by aircraft during landing and takeoff. Over the years that followed, research was carried out at NASA aeronautics research centers, often in collaboration with academia and industry, addressing the problem. In 2013, a high-level milestone was established, described as a Technical Challenge, with the objective of demonstrating the feasibility of a low-boom supersonic airliner that could meet current airport noise regulations. The Technical Challenge was formally called a Low Noise Propulsion for Low Boom Aircraft and was completed in late 2016. This paper reports the technical findings from this Technical Challenge, reaching back almost 10 years to review the technologies and tools that were developed along the way. It also discusses the final aircraft configuration and propulsion systems required for a supersonic civilian aircraft to meet noise regulations using the technologies available today. Finally, the paper documents the model-scale tests that validated the acoustic performance of the study aircraft.

Final Revisions Rule State Budgets and New Unit Set-Asides TSD

EPA Pesticide Factsheets

This technical support document shows the underlying data and calculations used to quantify the state budget revisions and new unit set-aside revisions made in the final revisions rule, as well as those revisions included in the direct final revisions rule

Medical Surveillance Monthly Report (MSMR). Volume 11, Number 2, April 2005

DTIC Science & Technology

2005-04-01

other year since 2000 (Tables 1,2,3). Finally, in 2004, there were nine reports of Rocky Mountain spotted fever , four of Reportable Medical Events...1 1 5 . 1 Rift Valley fever . . . . . Diphtheria . . . . . Rocky Mountain spotted fever 12... Rocky Mountain spotted fever . . . . 3 Escherichia coli O157:H7

BIOSENSOR FOR DIRECT DETERMINATION OF ORGANOPHOSPHATE NERVE AGENTS USING RECOMBINANT ESCHERICHIA COLI WITH SURFACE-EXPRESSED ORGANOPHOSPHORUS HYDROLASE. 1. POTENTIOMETRIC MICROBIAL ELECTRODE. (R823663)

EPA Science Inventory

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

Correction to: Top Down Tandem Mass Spectrometric Analysis of a Chemically Modified Rough-Type Lipopolysaccharide Vaccine Candidate

NASA Astrophysics Data System (ADS)

Oyler, Benjamin L.; Khan, Mohd M.; Smith, Donald F.; Harberts, Erin M.; Kilgour, David P. A.; Ernst, Robert K.; Cross, Alan S.; Goodlett, David R.

2018-04-01

In the preceding article "Top Down Tandem Mass Spectrometric Analysis of a Chemically Modified Rough-Type Lipopolysaccharide Vaccine Candidate" by Oyler et al., an error in the J5 E. coli LPS chemical structure (Figs. 2 and 4) was introduced and propagated into the final revision.

Coexpression of interleukin-6 and -2 from giant panda in Escherichia coli and the biological activity of the fusion protein.

PubMed

Yi, Y; Nian, Y-Y; Ji, H-W; Zhang, H; Zhu, L; Xu, Z-W

2013-06-14

To construct a fusion cytokine protein with more and stronger bioactivities to enhance the immunity of the cytokine alone, we expressed interleukin (IL)-6/(IL)-2 from giant panda (Ailuropoda melanoleuca) in Escherichia coli as a 59.4-kDa fusion protein. Subsequently, the inclusion bodies were solubilized with 8 M urea and applied onto a Ni-nitrilotriacetic acid column. The final production of IL-6/IL-2 reached 6 mg/L in soluble form, and the purified final product was >96% pure. In Western blot assays, the recombinant IL-6/IL-2 was recognized by polyclonal antibodies against IL-6 and IL-2 of giant panda. The results demonstrated that the protein mixture contained correctly folded IL-2 and IL-6 proteins. A 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide assay demonstrated that IL-6/IL-2 can promote lymphocyte proliferation and differentiation. These data suggest that the fusion protein could be used to develop a novel immunoadjuvant to enhance the immunity of animals against infectious diseases.

Immobilized Hydrolytic Enzymes Exhibit Antibiofilm Activity Against Escherichia coli at Sub-Lethal Concentrations.

PubMed

Villa, Federica; Secundo, Francesco; Polo, Andrea; Cappitelli, Francesca

2015-07-01

The effects of two commercially available immobilized enzymes (namely the glycosidase pectinase and the protease subtilisin A) at sub-lethal concentrations were investigated in terms of their influence on biofilm genesis, on the composition of the biofilm matrix, and their antibiotic synergy against Escherichia coli biofilm, used as a model system of bacterial biofilms. The best antibiofilm performance of solid-supported hydrolases was obtained at the surface concentration of 0.022 and 0.095 U/cm(2) with a reduction of 1.2 and 2.3 log CFU/biofilm for pectinase and subtilisin, respectively. At these enzyme surface concentrations, the biocatalysts affected the structural composition of the biofilm matrix, impacting biofilm thickness. Finally, the immobilized hydrolases enhanced biofilm sensitivity to a clinically relevant concentration of the antibiotic ampicillin. At the final antibiotic concentration of 0.1 mg/ml, a reduction of 2 and 3.5 log10 units in presence of 0.022 Upectinase/cm(2) and 0.095 Usubtilisin/cm(2) was obtained, respectively, in comparison the antibiotic alone. Immobilized pectinase and subtilisin at sub-lethal concentrations demonstrated a great potential for antibiofilm applications.

77 FR 37284 - Technical Amendments

Federal Register 2010, 2011, 2012, 2013, 2014

2012-06-21

... DEPARTMENT OF LABOR Office of Workers' Compensation Programs 20 CFR Parts 701, 702, 703, 725, and 726 RIN 1240-AA05 Technical Amendments AGENCY: Office of Workers' Compensation Programs, Labor. ACTION: Final rule. SUMMARY: The Office of Workers' Compensation Programs is making [[Page 37285

Vehicle infrastructure integration proof of concept : technical description--vehicle : final report

DOT National Transportation Integrated Search

2009-05-19

This report provides the technical description of the VII system developed for the Cooperative Agreement VII Program between the USDOT and the VII Consortium. The basic architectural elements are summarized and detailed descriptions of the hardware a...

FDA Escherichia coli Identification (FDA-ECID) Microarray: a Pangenome Molecular Toolbox for Serotyping, Virulence Profiling, Molecular Epidemiology, and Phylogeny

PubMed Central

Patel, Isha R.; Gangiredla, Jayanthi; Lacher, David W.; Mammel, Mark K.; Jackson, Scott A.; Lampel, Keith A.

2016-01-01

ABSTRACT Most Escherichia coli strains are nonpathogenic. However, for clinical diagnosis and food safety analysis, current identification methods for pathogenic E. coli either are time-consuming and/or provide limited information. Here, we utilized a custom DNA microarray with informative genetic features extracted from 368 sequence sets for rapid and high-throughput pathogen identification. The FDA Escherichia coli Identification (FDA-ECID) platform contains three sets of molecularly informative features that together stratify strain identification and relatedness. First, 53 known flagellin alleles, 103 alleles of wzx and wzy, and 5 alleles of wzm provide molecular serotyping utility. Second, 41,932 probe sets representing the pan-genome of E. coli provide strain-level gene content information. Third, approximately 125,000 single nucleotide polymorphisms (SNPs) of available whole-genome sequences (WGS) were distilled to 9,984 SNPs capable of recapitulating the E. coli phylogeny. We analyzed 103 diverse E. coli strains with available WGS data, including those associated with past foodborne illnesses, to determine robustness and accuracy. The array was able to accurately identify the molecular O and H serotypes, potentially correcting serological failures and providing better resolution for H-nontypeable/nonmotile phenotypes. In addition, molecular risk assessment was possible with key virulence marker identifications. Epidemiologically, each strain had a unique comparative genomic fingerprint that was extended to an additional 507 food and clinical isolates. Finally, a 99.7% phylogenetic concordance was established between microarray analysis and WGS using SNP-level data for advanced genome typing. Our study demonstrates FDA-ECID as a powerful tool for epidemiology and molecular risk assessment with the capacity to profile the global landscape and diversity of E. coli. IMPORTANCE This study describes a robust, state-of-the-art platform developed from available whole-genome sequences of E. coli and Shigella spp. by distilling useful signatures for epidemiology and molecular risk assessment into one assay. The FDA-ECID microarray contains features that enable comprehensive molecular serotyping and virulence profiling along with genome-scale genotyping and SNP analysis. Hence, it is a molecular toolbox that stratifies strain identification and pathogenic potential in the contexts of epidemiology and phylogeny. We applied this tool to strains from food, environmental, and clinical sources, resulting in significantly greater phylogenetic and strain-specific resolution than previously reported for available typing methods. PMID:27037122

Utilization of high temperature compost in space agriculture: the model compost kills Escherichia coli

NASA Astrophysics Data System (ADS)

Oshima, Tairo; Moriya, Toshiyuki; Yoshii, Takahiro

The author and his colleagues have proposed the use of high temperature composting in space inhabitation. Composting has many advantages over burning in organic waste treatments. Composting is self-heating processes and needs no extra fuel. Composting requires no sophis-ticated equipment such as an incinerator. Composting emits no hazardous gases such as NOx, SOx and dioxines which are often produced by burning. The final product can be used as fer-tilizer in space farm land; resources recycling society can be constructed in space stations and space cities. In addition to these advantages, composting and compost soil may contribute to the environmental cleanup. During composting processes, harmful compounds to agricultural plants and animals can be destroyed. Seeds of weeds can be killed by high heat. Likewise pathogenic microbes in the waste can be eliminated during fermentation inside the composts. Recently we measured the survivability of E. coli in compost. E. coli was used as the represen-tative of the Gram-negative bacteria. Since many pathogenic strains belong to Gram-negative bacteria and Gram-negative bacteria are more resistant to antibiotics than gram-positive bac-teria. When E. coli cells were mixed in the compost pile of which inside temperature reaches up to 75oC, they died within a short period as expected. However, E. coli DNA was detected even after a day in high temperature compost. RNA has a shorter life-span than DNA, but was detected after incubation in compost for several hours. In addition to sterilizing effects due to high temperature, we found our compost soil has E. coli killing activity. When mixed with the compost soil at room temperature, E. coli died gradually. Extract of the compost soil also killed E. coli at room temperature, but it took a few days to eliminate E. coli completely. During the killing process, total number of living bacteria did not change, indicating that the killing activity is limited to some specific microorganisms. These findings suggest that the compost can be used to eliminate some of deleterious microbes from the environment without damages to the beneficial microbes. We are planning to test the killing activity of the com-post soil against more dangerous microorganisms such as Salmonella species, especially those pathogenic to barn animals.

NetView technical research

NASA Technical Reports Server (NTRS)

1993-01-01

This is the Final Technical Report for the NetView Technical Research task. This report is prepared in accordance with Contract Data Requirements List (CDRL) item A002. NetView assistance was provided and details are presented under the following headings: NetView Management Systems (NMS) project tasks; WBAFB IBM 3090; WPAFB AMDAHL; WPAFB IBM 3084; Hill AFB; McClellan AFB AMDAHL; McClellan AFB IBM 3090; and Warner-Robins AFB.

Inventory of greenhouse gas emissions from on-road vehicles in Midwestern USA States and integrated approach to achieving environmental sustainability in transportation : USDOT Region V Regional University Transportation Center final report : technical su

DOT National Transportation Integrated Search

2016-12-29

Two project objectives one technical and one educational- were laid out in this project. The technical objective was to assess current inventory of greenhouse gases (GHG) in the six Midwestern states of the nation and to estimate improvements as ...

Emissions Inventory Final Rule TSD

EPA Pesticide Factsheets

This technical support document (TSD) provides the details of emissions data processing done in support of EPA's final rulemaking effort for the Federal Transport Rule, now known as the Cross-State Air Pollution Rule.

Intestinal-borne dermatoses significantly improved by oral application of Escherichia coli Nissle 1917.

PubMed

Manzhalii, Elina; Hornuss, Daniel; Stremmel, Wolfgang

2016-06-21

To evaluate the effect of oral Escherichia coli (E. coli) Nissle application on the outcome of intestinal-borne dermatoses. In a randomized, controlled, non-blinded prospective clinical trial 82 patients with intestinal-borne facial dermatoses characterized by an erythematous papular-pustular rash were screened. At the initiation visit 37 patients entered the experimental arm and 20 patients constituted the control arm. All 57 patients were treated with a vegetarian diet and conventional topical therapy of the dermatoses with ointments containing tetracycline, steroids and retinoids. In the experimental arm patients received a one month therapy with oral E. coli Nissle at a maintenance dose of 2 capsules daily. The experimental group was compared to a non-treatment group only receiving the diet and topical therapy. The primary outcome parameter was improvement of the dermatoses, secondary parameters included life quality and adverse events. In addition the immunological reaction profile (IgA, interleucin-8 and interferon-α) was determined. Furthermore the changes of stool consistency and the microbiota composition over the time of intervention were recorded. Eighty-nine percent of the patients with acne, papular-pustular rosacea and seborrhoic dermatitis responded to E. coli Nissle therapy with significant amelioration or complete recovery in contrast to 56% in the control arm (P < 0.01). Accordingly, in the E. coli Nissle treated patients life quality improved significantly (P < 0.01), and adverse events were not recorded. The clinical improvement was associated with a significant increase of IgA levels to normal values in serum as well as suppression of the proinflammatory cytokine IL-8 (P < 0.01 for both parameters). In the E. coli Nissle treated group a shift towards a protective microbiota with predominance of bifidobacteria and lactobacteria (> 10(7) CFU/g stool) was observed in 79% and 63% of the patients, respectively (P < 0.01), compared to no change in the control group without E. coli Nissle. Moreover, the detection rate of a pathogenic flora dropped from 73% to 14 % of the patients in the experimental arm (P < 0.01) with no significant change in the control arm (accounting 80% before and 70% after the observation period, P > 0.05). Accordingly, stool consistency, color and smell normalized in the E. coli Nissle treated patients. E. coli Nissle protects the mucus barrier by overgrowth of a favorable gut microbiota with less immunoreactive potential which finally leads to clinical improvement of intestinal borne dermatoses.

Transportation Network Data Requirements for Assessing Criticality for Resiliency and Adaptation Planning

DOT National Transportation Integrated Search

2017-11-01

This report is one of two NCST Research Report documents produced as part of a project to advance the technical modeling tools for resiliency and adaptation planning, especially those used for criticality rankings. The official final technical report...

Comment Deadlines Established Regarding the LightSquared Technical Working Group Report

DOT National Transportation Integrated Search

2011-06-30

On June 30, 2011, LightSquared Subsidiary LLC (LightSquared) submitted a final report of the : technical working group co-chaired by LightSquared and the United States Global Positioning System : (GPS) Industry Council (USGIC)1 and organized in respo...

Technical report on prototype intelligent network flow optimization (INFLO) dynamic speed harmonization and queue warning.

DOT National Transportation Integrated Search

2015-06-01

This Technical Report on Prototype Intelligent Network Flow Optimization (INFLO) Dynamic Speed Harmonization and Queue Warning is the final report for the project. It describes the prototyping, acceptance testing and small-scale demonstration of the ...

NUSC Technical Publications Guide.

DTIC Science & Technology

1985-05-01

Facility personnel especially that of A. Castelluzzo, E. Deland, J. Gesel , and E. Szlosek (all of Code 4343). Reviewed and Approved: 14 July 1980 D...their technical content and format. Review and approve the manual outline, the review manuscript, and the final camera - reproducible copy. Conduct in

Cross-State Air Pollution Rule Update Allowance Allocation Final Rule TSD

EPA Pesticide Factsheets

This Technical Support Document (TSD) provides information that supports EPA’s determination of unit-level allocations for existing and new units under the final Cross-State Air Pollution Rule Update.

NMSBA: Aken Technologies Final Report: Toxicity Testing of Liquidoff

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ruffing, Anne; Jensen, Travis; Strickland, Lucas

2015-02-01

To determine the effect of Liquidoff on bacteria, three bacterial strains were tested: Escherichia coli DH5α, Synechococcus sp. PCC 7002, and Synechococcus elongatus PCC 7942. E. coli DH5α is a Gram-negative, aerobic bacterium that is often found in normal gut flora and is commonly used the laboratory due to its fast growth rate. Synechococcus sp. PCC 7002 and S. elongatus PCC 7942 are Gram-negative, aquatic, autophototrophic cyanobacteria. Synechococcus sp. PCC 7002 is a marine cyanobacterium isolated from ‘fish pens’ on Magueyes Island, Puerto Rico in 1962, while S. elongatus PCC 7942 is a freshwater cyanobacterium. It should be noted thatmore » no Gram-positive bacterium was tested in this study.« less

Dissection and engineering of the Escherichia coli formate hydrogenlyase complex.

PubMed

McDowall, Jennifer S; Hjersing, M Charlotte; Palmer, Tracy; Sargent, Frank

2015-10-07

The Escherichia coli formate hydrogenlyase (FHL) complex is produced under fermentative conditions and couples formate oxidation to hydrogen production. In this work, the architecture of FHL has been probed by analysing affinity-tagged complexes from various genetic backgrounds. In a successful attempt to stabilize the complex, a strain encoding a fusion between FdhF and HycB has been engineered and characterised. Finally, site-directed mutagenesis of the hycG gene was performed, which is predicted to encode a hydrogenase subunit important for regulating sensitivity to oxygen. This work helps to define the core components of FHL and provides solutions to improving the stability of the enzyme. Copyright © 2015 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Guidelines for producing training films and videos.

PubMed

Harper, P B

1991-01-01

Drawing from experience in producing a film on the surgical procedure of female sterilization, 4 guidelines to technical film production for training purposes are presented and discussed in this paper. In order of presentation in the text, the paper 1st encourages identifying and securing a technical expert, then clearly identifying steps of the technical procedure, involving trainees and trainers in the production process, and working with experienced producers, scriptwriters, and crew members. Returning to the 1st guideline, the technical advisor will have a central presence during all photography and editing, and ideally should not have any personal investment in the procedure being shown. Prior to script finalization and sorting, research is urged to ensure concrete procedural steps. Printed materials, slides, interviews of experienced clinicians, procedure observation, and test videotape shooting may be called upon and employed as parts of the research phase. Trainees should participate during preliminary research, script development, and pretesting of early film versions, their suggestions for change incorporated where appropriate in the final version. On the final point of securing experienced workers, country nationals sensitive to relevant cultural and background dynamics should be included in the team. The special concerns of airport security regulation and customs requirements knowledge are essential, as well as their attention to assuring adequate on-site electricity for camera equipment.

Contamination Revealed by Indicator Microorganism Levels during Veal Processing.

PubMed

Bosilevac, Joseph M; Wang, Rong; Luedtke, Brandon E; Wheeler, Tommy L; Koohmaraie, Mohammad

2016-08-01

During site visits of veal processors, the U.S. Department of Agriculture, Food Safety Inspection Service (FSIS) has reported processing deficiencies that likely contribute to increased levels of veal contamination. Here, we report the results of measuring aerobic plate count bacteria (APC), Enterobacteriaceae, coliforms (CF), and Escherichia coli during eight sample collections at five veal processors to assess contamination during the harvest of bob veal and formula-fed veal before (n = 5 plants) and after (n = 3 plants) changes to interventions and processing practices. Hides of veal calves at each plant had mean log CFU/100 cm(2) APC, Enterobacteriaceae, CF, and E. coli of 6.02 to 8.07, 2.95 to 5.24, 3.28 to 5.83, and 3.08 to 5.59, respectively. Preintervention carcasses had mean log CFU/100 cm(2) APC, Enterobacteriaceae, CF, and E. coli of 3.08 to 5.22, 1.16 to 3.47, 0.21 to 3.06, and -0.07 to 3.10, respectively, before and 2.72 to 4.50, 0.99 to 2.76, 0.69 to 2.26, and 0.33 to 2.12, respectively, after changes were made to improve sanitary dressing procedures. Final veal carcasses had mean log CFU/100 cm(2) APC, Enterobacteriaceae, CF, and E. coli of 0.36 to 2.84, -0.21 to 1.59, -0.23 to 1.59, and -0.38 to 1.45 before and 0.44 to 2.64, -0.16 to 1.33, -0.42 to 1.20, and 0.48 to 1.09 after changes were made to improve carcass-directed interventions. Whereas the improved dressing procedures resulted in improved carcass cleanliness, the changes to carcass-directed interventions were less successful, and veal processors are urged to use techniques that ensure uniform and consistent delivery of antimicrobials to carcasses. Analysis of results comparing bob veal to formula-fed veal found bob veal hides, preintervention carcasses, and final carcasses to have increased (P < 0.05) APC, Enterobacteriaceae, CF, and E. coli (with the exception of hide Enterobacteriaceae; P > 0.05) relative to formula fed veal. When both veal categories were harvested at the same plant on the same day, similar results were observed. Since identification by FSIS, the control of contamination during veal processing has started to improve, but challenges still persist.

Oral antimicrobials increase antimicrobial resistance in porcine E. coli--a systematic review.

PubMed

Burow, E; Simoneit, C; Tenhagen, B-A; Käsbohrer, A

2014-03-01

Administration of antimicrobials to livestock increases the risk of antimicrobial resistance (AMR) in commensal bacteria. Antimicrobials in pig production are usually administered per pen via feed which implies treatment of sick alongside with healthy animals. The objective of this systematic literature review was to investigate the effect of orally administered antimicrobials on AMR in Escherichia coli of swine. Studies published in peer reviewed journals were retrieved from the international online databases ISI Web of Knowledge, PubMed, Scopus and the national electronic literature data base of Deutsches Institut für Medizinische Dokumentation und Information. The studies were assessed using the eligibility criteria English or German language, access to full paper version, defined treatment and control group (initial value or non-treatment) as well as administration and resistance testing of the same antimicrobial class. In the qualitative synthesis, only studies were included presenting the summary measures odds ratio or prevalence of resistance, the category of the applied antimicrobial and the dosage. An effect of the antimicrobial on AMR in E. coli was evaluated as an "increase", "no effect" or "decrease" if the odds or alternatively the prevalence ratio were >1.0, 1.0 or <1.0, respectively. Eleven studies, describing 36 different trials, fulfilled the eligibility criteria and were finally assessed. An increase of AMR in E. coli was found in 10 out of 11 trials comparing AMR after with AMR prior to oral treatment and in 22 of the 25 trials comparing orally treated with untreated groups. Effects expressed as odds or prevalence ratios were highest for the use of aminoglycosides, quinolones and tetracycline. There was no clear association between the reported dosage and AMR towards tetracycline. Information on antimicrobial substance and dosage was missing in 4 and 5 of the 11 finally selected studies. The 36 identified trials were inhomogenous in usage and provision of information on sample size. Oral administration of antimicrobials increases the risk of AMR in E. coli from swine. There is however a lack of studies on the impact of dosage and longitudinal effects of treatment. The published studies have a number of issues concerning their scientific quality. More high quality research is needed to better address and quantifiy the effect of orally administered antimicrobials on AMR in swine. Copyright © 2014 Elsevier B.V. All rights reserved.

Northeast Corridor Improvement Project Electrification - New Haven, CT to Boston, MA : Final Environmental Statement/Report : v. 2. Technical Studies

DOT National Transportation Integrated Search

1994-10-31

This document is the final environmental impact statement and final environmental impact report (FEIS/R) on the proposal by the National Railroad Passenger Corporation (Amtrak) to complete the electrification of the Northeast Corridor main line by ex...

Health care fraud and abuse data collection program: technical revisions to Healthcare Integrity and Protection Data Bank data collection activities. Final rule.

PubMed

2004-09-21

The rule finalizes technical changes to the Healthcare Integrity and Protection Data Bank (HIPDB) data collection reporting requirements by clarifying the types of personal numeric identifiers that may be reported to the data bank in connection with adverse actions. The rule clarifies that in lieu of a Social Security Number (SSN), an individual taxpayer identification number (ITIN) may be reported to the data bank when, in those limited situations, an individual does not have an SSN.

Recovery Act-SmartGrid regional demonstration transmission and distribution (T&D) Infrastructure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hedges, Edward T.

This document represents the Final Technical Report for the Kansas City Power & Light Company (KCP&L) Green Impact Zone SmartGrid Demonstration Project (SGDP). The KCP&L project is partially funded by Department of Energy (DOE) Regional Smart Grid Demonstration Project cooperative agreement DE-OE0000221 in the Transmission and Distribution Infrastructure application area. This Final Technical Report summarizes the KCP&L SGDP as of April 30, 2015 and includes summaries of the project design, implementation, operations, and analysis performed as of that date.

Final Technical Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sobecky, Patricia A; Taillefert, Martial

This final technical report describes results and findings from a research project to examine the role of microbial phosphohydrolase enzymes in naturally occurring subsurface microorganisms for the purpose of promoting the immobilization of the radionuclide uranium through the production of insoluble uranium phosphate minerals. The research project investigated the microbial mechanisms and the physical and chemical processes promoting uranium biomineralization and sequestration in oxygenated subsurface soils. Uranium biomineralization under aerobic conditions can provide a secondary biobarrier strategy to immobilize radionuclides should the metal precipitates formed by microbial dissimilatory mechanisms remobilize due to a change in redox state.

Waveguide Studies for Fiber Optics and Optical Signal Processing Applications.

DTIC Science & Technology

1980-04-01

AO-A086 115 UNI!VERtSIT? OF SOUTIUR CALEPCRNA LOS AMUSS / 5 WAVGUIDE STUIES15 FOR FEB53 OpTECS AND OpTICAL SEOSA.o P /0Ksu-y "/6 UNLSIIDAPR N0 E...SAMUE Flola-??-c-sa UNCASZFIORAC-M-8042 U Final Technical Report (1 1April 1950 L V ~ WAVEGUIDE STUDIES FOR FIBER OPTICS AND OPTICAL SIGNAL PROCESSING...and Subtitle) 081 6&4𔃾JODO )EI YAVECUIDESTUDIES FOR JIBER OPTICS ANDL 7 Final ,T/echnical epoErt, OPTICAL SI’tNAL PROCESSING APPLICATIONS.4 11 Se 77

SELECTIVE DISSEMINATION OF INFORMATION (SDI). VOLUME I, PILOT TEST AT U.S. ARMY NATICK LABORATORIES. FINAL REPORT. ARMY TECHNICAL LIBRARY IMPROVEMENT STUDIES (ATLIS), REPORT NO. 15.

ERIC Educational Resources Information Center

BIVONA, WILLIAM A.

THIS VOLUME PRESENTS THE RESULTS OF A NINE-MONTH TEST OF A PROTOTYPE SELECTIVE DISSEMINATION OF INFORMATION (SDI) SYSTEM DEVELOPED FOR THE ARMY TECHNICAL LIBRARIES. DURING THE PILOT TEST ONE THOUSAND DOCUMENTS WERE CATALOGED, INDEXED, AND DISSEMINATED TO TWENTY-FIVE SCIENTIFIC AND TECHNICAL PERSONNEL. MATCHING OF THE INTEREST PROFILES OF THESE…

Fate of Shiga toxin-producing O157:H7 and non-O157:H7 Escherichia coli cells within refrigerated, frozen, or frozen then thawed ground beef patties cooked on a commercial open-flame gas or a clamshell electric grill.

PubMed

Luchansky, John B; Porto-Fett, Anna C S; Shoyer, Bradley A; Phillips, John; Chen, Vivian; Eblen, Denise R; Cook, L Victor; Mohr, Tim B; Esteban, Emilio; Bauer, Nathan

2013-09-01

Both high-fat and low-fat ground beef (percent lean:fat = ca. 70:30 and 93:7, respectively) were inoculated with a 6-strain cocktail of non-O157:H7 Shiga toxin-producing Escherichia coli (STEC) or a five-strain cocktail of E. coli O157:H7 (ca. 7.0 log CFU/g). Patties were pressed (ca. 2.54 cm thick, ca. 300 g each) and then refrigerated (4°C, 18 to 24 h), or frozen (-18°C, 3 weeks), or frozen (-18°C, 3 weeks) and then thawed (4°C for 18 h or 21°C for 10 h) before being cooked on commercial gas or electric grills to internal temperatures of 60 to 76.6°C. For E. coli O157:H7, regardless of grill type or fat level, cooking refrigerated patties to 71.1 or 76.6°C decreased E. coli O157:H7 numbers from an initial level of ca. 7.0 log CFU/g to a final level of ≤1.0 log CFU/g, whereas decreases to ca. 1.1 to 3.1 log CFU/g were observed when refrigerated patties were cooked to 60.0 or 65.5°C. For patties that were frozen or freeze-thawed and cooked to 71.1 or 76.6°C, E. coli O157:H7 numbers decreased to ca. 1.7 or ≤0.7 log CFU/g. Likewise, pathogen numbers decreased to ca. 0.7 to 3.7 log CFU/g in patties that were frozen or freeze-thawed and cooked to 60.0 or 65.5°C. For STEC, regardless of grill type or fat level, cooking refrigerated patties to 71.1 or 76.6°C decreased pathogen numbers from ca. 7.0 to ≤0.7 log CFU/g, whereas decreases to ca. 0.7 to 3.6 log CFU/g were observed when refrigerated patties were cooked to 60.0 or 65.5°C. For patties that were frozen or freeze-thawed and cooked to 71.1 or 76.6°C, STEC numbers decreased to a final level of ca. 1.5 to ≤0.7 log CFU/g. Likewise, pathogen numbers decreased from ca. 7.0 to ca. 0.8 to 4.3 log CFU/g in patties that were frozen or freeze-thawed and cooked to 60.0 or 65.5°C. Thus, cooking ground beef patties that were refrigerated, frozen, or freeze-thawed to internal temperatures of 71.1 and 76.6°C was effective for eliminating ca. 5.1 to 7.0 log CFU of E. coli O157:H7 and STEC per g.

Biofilm Formation and Dispersal under the Influence of the Global Regulator CsrA of Escherichia coli

PubMed Central

Jackson, Debra W.; Suzuki, Kazushi; Oakford, Lawrence; Simecka, Jerry W.; Hart, Mark E.; Romeo, Tony

2002-01-01

The predominant mode of growth of bacteria in the environment is within sessile, matrix-enclosed communities known as biofilms. Biofilms often complicate chronic and difficult-to-treat infections by protecting bacteria from the immune system, decreasing antibiotic efficacy, and dispersing planktonic cells to distant body sites. While the biology of bacterial biofilms has become a major focus of microbial research, the regulatory mechanisms of biofilm development remain poorly defined and those of dispersal are unknown. Here we establish that the RNA binding global regulatory protein CsrA (carbon storage regulator) of Escherichia coli K-12 serves as both a repressor of biofilm formation and an activator of biofilm dispersal under a variety of culture conditions. Ectopic expression of the E. coli K-12 csrA gene repressed biofilm formation by related bacterial pathogens. A csrA knockout mutation enhanced biofilm formation in E. coli strains that were defective for extracellular, surface, or regulatory factors previously implicated in biofilm formation. In contrast, this csrA mutation did not affect biofilm formation by a glgA (glycogen synthase) knockout mutant. Complementation studies with glg genes provided further genetic evidence that the effects of CsrA on biofilm formation are mediated largely through the regulation of intracellular glycogen biosynthesis and catabolism. Finally, the expression of a chromosomally encoded csrA′-′lacZ translational fusion was dynamically regulated during biofilm formation in a pattern consistent with its role as a repressor. We propose that global regulation of central carbon flux by CsrA is an extremely important feature of E. coli biofilm development. PMID:11741870

Evaluation of different micro/nanobeads used as amplifiers in QCM immunosensor for more sensitive detection of E. coli O157:H7.

PubMed

Jiang, Xuesong; Wang, Ronghui; Wang, Yun; Su, Xiaoli; Ying, Yibin; Wang, Jianping; Li, Yanbin

2011-11-15

Micro/nanobeads with different materials (magnetic, silica and polymer) and different sizes (diameters from 30nm to 970nm) were investigated for their use as amplifiers in a quartz crystal microbalance (QCM) immunosensor for more sensitive detection of Escherichia coli O157:H7. The micro/nanobeads were conjugated with anti-E. coli antibodies. E. coli O157:H7 cells were first captured by the first antibody immobilized on the electrode surface, and then micro/nanobeads labeled secondary antibodies attached to the cells, and finally the complexes of antibody-E. coli-antibody modified beads were formed. The results showed that antibody-labeled beads lead to signal amplification in both the change in frequency (ΔF) and the change in resistance (ΔR). Since the penetration depth of the oscillation-induced shear-waves for a ∼8MHz crystal is limited to 200nm, the interpretation of how the signal is amplified by the adsorbed particles was represented in terms of the coupled-oscillator theory. The amplification is not sensed in terms of increase in mass on the sensor surface. Amplification is sensed as a change in bacterial resonance frequency when the spheres adsorb to the bacteria. The change in the values of ΔF caused by different micro/nanobeads (amplifiers) attaching on target bacterial cells is indicative of the ratio between the resonance frequency of the absorbed bacterial-particle complex (ω(s)), and the resonance frequency of the crystal (ω). Copyright © 2011 Elsevier B.V. All rights reserved.

Novel MntR-Independent Mechanism of Manganese Homeostasis in Escherichia coli by the Ribosome-Associated Protein HflX

PubMed Central

Kaur, Gursharan; Sengupta, Sandeepan; Kumar, Vineet; Kumari, Aruna; Ghosh, Aditi; Parrack, Pradeep

2014-01-01

Manganese is a micronutrient required for activities of several important enzymes under conditions of oxidative stress and iron starvation. In Escherichia coli, the manganese homeostasis network primarily constitutes a manganese importer (MntH) and an exporter (MntP), which are regulated by the MntR dual regulator. In this study, we find that deletion of E. coli hflX, which encodes a ribosome-associated GTPase with unknown function, renders extreme manganese sensitivity characterized by arrested cell growth, filamentation, lower rate of replication, and DNA damage. We demonstrate that perturbation by manganese induces unprecedented influx of manganese in ΔhflX cells compared to that in the wild-type E. coli strain. Interestingly, our study indicates that the imbalance in manganese homeostasis in the ΔhflX strain is independent of the MntR regulon. Moreover, the influx of manganese leads to a simultaneous influx of zinc and inhibition of iron import in ΔhflX cells. In order to review a possible link of HflX with the λ phage life cycle, we performed a lysis-lysogeny assay to show that the Mn-perturbed ΔhflX strain reduces the frequency of lysogenization of the phage. This observation raises the possibility that the induced zinc influx in the manganese-perturbed ΔhflX strain stimulates the activity of the zinc-metalloprotease HflB, the key determinant of the lysis-lysogeny switch. Finally, we propose that manganese-mediated autophosphorylation of HflX plays a central role in manganese, zinc, and iron homeostasis in E. coli cells. PMID:24794564

Ceramic High Efficiency Particulate Air (HEPA) Filter Final Report CRADA No. TC02160.0

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mitchell, M.; Bergman, W.

2017-08-25

The technical objective of this project was to develop a ceramic HEPA filter technology, by initially producing and testing coupon ceramics, small scale prototypes, and full scale prototype HEPA filters, and to address relevant manufacturing and commercialization technical issues.

7 CFR 614.6 - Agency records and decision notices.

Code of Federal Regulations, 2010 CFR

2010-01-01

... notifies participants of the agency's preliminary and final technical determinations and program decisions... decision notice within 10 working days of rendering a technical determination or program decision. In lieu of certified mail, NRCS may hand deliver notices to participants with written acknowledgment of...

ECOS E-MATRIX Methane and Volatile Organic Carbon (VOC) Emissions Best Practices Database

DOE Office of Scientific and Technical Information (OSTI.GOV)

Parisien, Lia

2016-01-31

This final scientific/technical report on the ECOS e-MATRIX Methane and Volatile Organic Carbon (VOC) Emissions Best Practices Database provides a disclaimer and acknowledgement, table of contents, executive summary, description of project activities, and briefing/technical presentation link.

77 FR 18914 - National Motor Vehicle Title Information System (NMVTIS): Technical Corrections

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-29

... 1121-AA79 National Motor Vehicle Title Information System (NMVTIS): Technical Corrections AGENCY... (OJP) is promulgating this direct final rule for its National Motor Vehicle Title Information System... INFORMATION CONTACT paragraph. II. Background The National Motor Vehicle Title Information System was...

Intelligent transportation system (ITS) study for the Buffalo and Niagara Falls metropolitan area, Erie and Niagara Counties, New York : final report, technical summary

DOT National Transportation Integrated Search

1997-06-18

This document provides a technical summary for the seven working papers prepared for the New York State Department of Transportation (NYSDOT) Buffalo and Niagara Falls Intelligent Transportation System (ITS) Study.

77 FR 8095 - Technical Corrections to Commission Regulations

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-14

... DEPARTMENT OF ENERGY Federal Energy Regulatory Commission 18 CFR Part 2 [Docket No. RM11-30-000; Order No. 756] Technical Corrections to Commission Regulations Issued February 8, 2012. AGENCY: Federal Energy Regulatory Commission, DOE. ACTION: Final rule: correcting amendment. SUMMARY: This document adds...

78 FR 13543 - Defense Federal Acquisition Regulation Supplement; Technical Amendments

Federal Register 2010, 2011, 2012, 2013, 2014

2013-02-28

... DEPARTMENT OF DEFENSE Defense Acquisition Regulations System 48 CFR Parts 201, 204, 215, 225, 227, 242, 245, and 252 Defense Federal Acquisition Regulation Supplement; Technical Amendments AGENCY: Defense Acquisition Regulations System, Department of Defense (DoD). ACTION: Final rule. SUMMARY: DoD is...

Validation on milk and sprouts of EN ISO 16654:2001 - Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Escherichia coli O157.

PubMed

Tozzoli, Rosangela; Maugliani, Antonella; Michelacci, Valeria; Minelli, Fabio; Caprioli, Alfredo; Morabito, Stefano

2018-05-08

In 2006, the European Committee for standardisation (CEN)/Technical Committee 275 - Food analysis - Horizontal methods/Working Group 6 - Microbiology of the food chain (TC275/WG6), launched the project of validating the method ISO 16654:2001 for the detection of Escherichia coli O157 in foodstuff by the evaluation of its performance, in terms of sensitivity and specificity, through collaborative studies. Previously, a validation study had been conducted to assess the performance of the Method No 164 developed by the Nordic Committee for Food Analysis (NMKL), which aims at detecting E. coli O157 in food as well, and is based on a procedure equivalent to that of the ISO 16654:2001 standard. Therefore, CEN established that the validation data obtained for the NMKL Method 164 could be exploited for the ISO 16654:2001 validation project, integrated with new data obtained through two additional interlaboratory studies on milk and sprouts, run in the framework of the CEN mandate No. M381. The ISO 16654:2001 validation project was led by the European Union Reference Laboratory for Escherichia coli including VTEC (EURL-VTEC), which organized the collaborative validation study on milk in 2012 with 15 participating laboratories and that on sprouts in 2014, with 14 participating laboratories. In both studies, a total of 24 samples were tested by each laboratory. Test materials were spiked with different concentration of E. coli O157 and the 24 samples corresponded to eight replicates of three levels of contamination: zero, low and high spiking level. The results submitted by the participating laboratories were analyzed to evaluate the sensitivity and specificity of the ISO 16654:2001 method when applied to milk and sprouts. The performance characteristics calculated on the data of the collaborative validation studies run under the CEN mandate No. M381 returned sensitivity and specificity of 100% and 94.4%, respectively for the milk study. As for sprouts matrix, the sensitivity resulted in 75.9% in the low level of contamination samples and 96.4% in samples spiked with high level of E. coli O157 and specificity was calculated as 99.1%. Copyright © 2018 Elsevier B.V. All rights reserved.

Comparison of Metabolic Pathways in Escherichia coli by Using Genetic Algorithms.

PubMed

Ortegon, Patricia; Poot-Hernández, Augusto C; Perez-Rueda, Ernesto; Rodriguez-Vazquez, Katya

2015-01-01

In order to understand how cellular metabolism has taken its modern form, the conservation and variations between metabolic pathways were evaluated by using a genetic algorithm (GA). The GA approach considered information on the complete metabolism of the bacterium Escherichia coli K-12, as deposited in the KEGG database, and the enzymes belonging to a particular pathway were transformed into enzymatic step sequences by using the breadth-first search algorithm. These sequences represent contiguous enzymes linked to each other, based on their catalytic activities as they are encoded in the Enzyme Commission numbers. In a posterior step, these sequences were compared using a GA in an all-against-all (pairwise comparisons) approach. Individual reactions were chosen based on their measure of fitness to act as parents of offspring, which constitute the new generation. The sequences compared were used to construct a similarity matrix (of fitness values) that was then considered to be clustered by using a k-medoids algorithm. A total of 34 clusters of conserved reactions were obtained, and their sequences were finally aligned with a multiple-sequence alignment GA optimized to align all the reaction sequences included in each group or cluster. From these comparisons, maps associated with the metabolism of similar compounds also contained similar enzymatic step sequences, reinforcing the Patchwork Model for the evolution of metabolism in E. coli K-12, an observation that can be expanded to other organisms, for which there is metabolism information. Finally, our mapping of these reactions is discussed, with illustrations from a particular case.

Comparison of Metabolic Pathways in Escherichia coli by Using Genetic Algorithms

PubMed Central

Ortegon, Patricia; Poot-Hernández, Augusto C.; Perez-Rueda, Ernesto; Rodriguez-Vazquez, Katya

2015-01-01

In order to understand how cellular metabolism has taken its modern form, the conservation and variations between metabolic pathways were evaluated by using a genetic algorithm (GA). The GA approach considered information on the complete metabolism of the bacterium Escherichia coli K-12, as deposited in the KEGG database, and the enzymes belonging to a particular pathway were transformed into enzymatic step sequences by using the breadth-first search algorithm. These sequences represent contiguous enzymes linked to each other, based on their catalytic activities as they are encoded in the Enzyme Commission numbers. In a posterior step, these sequences were compared using a GA in an all-against-all (pairwise comparisons) approach. Individual reactions were chosen based on their measure of fitness to act as parents of offspring, which constitute the new generation. The sequences compared were used to construct a similarity matrix (of fitness values) that was then considered to be clustered by using a k-medoids algorithm. A total of 34 clusters of conserved reactions were obtained, and their sequences were finally aligned with a multiple-sequence alignment GA optimized to align all the reaction sequences included in each group or cluster. From these comparisons, maps associated with the metabolism of similar compounds also contained similar enzymatic step sequences, reinforcing the Patchwork Model for the evolution of metabolism in E. coli K-12, an observation that can be expanded to other organisms, for which there is metabolism information. Finally, our mapping of these reactions is discussed, with illustrations from a particular case. PMID:25973143

Phenotypic Changes Exhibited by E. coli Cultured in Space.

PubMed

Zea, Luis; Larsen, Michael; Estante, Frederico; Qvortrup, Klaus; Moeller, Ralf; Dias de Oliveira, Sílvia; Stodieck, Louis; Klaus, David

2017-01-01

Bacteria will accompany humans in our exploration of space, making it of importance to study their adaptation to the microgravity environment. To investigate potential phenotypic changes for bacteria grown in space, Escherichia coli was cultured onboard the International Space Station with matched controls on Earth. Samples were challenged with different concentrations of gentamicin sulfate to study the role of drug concentration on the dependent variables in the space environment. Analyses included assessments of final cell count, cell size, cell envelope thickness, cell ultrastructure, and culture morphology. A 13-fold increase in final cell count was observed in space with respect to the ground controls and the space flight cells were able to grow in the presence of normally inhibitory levels of gentamicin sulfate. Contrast light microscopy and focused ion beam/scanning electron microscopy showed that, on average, cells in space were 37% of the volume of their matched controls, which may alter the rate of molecule-cell interactions in a diffusion-limited mass transport regime as is expected to occur in microgravity. TEM imagery showed an increase in cell envelope thickness of between 25 and 43% in space with respect to the Earth control group. Outer membrane vesicles were observed on the spaceflight samples, but not on the Earth cultures. While E. coli suspension cultures on Earth were homogenously distributed throughout the liquid medium, in space they tended to form a cluster, leaving the surrounding medium visibly clear of cells. This cell aggregation behavior may be associated with enhanced biofilm formation observed in other spaceflight experiments.

Phenotypic Changes Exhibited by E. coli Cultured in Space

PubMed Central

Zea, Luis; Larsen, Michael; Estante, Frederico; Qvortrup, Klaus; Moeller, Ralf; Dias de Oliveira, Sílvia; Stodieck, Louis; Klaus, David

2017-01-01

Bacteria will accompany humans in our exploration of space, making it of importance to study their adaptation to the microgravity environment. To investigate potential phenotypic changes for bacteria grown in space, Escherichia coli was cultured onboard the International Space Station with matched controls on Earth. Samples were challenged with different concentrations of gentamicin sulfate to study the role of drug concentration on the dependent variables in the space environment. Analyses included assessments of final cell count, cell size, cell envelope thickness, cell ultrastructure, and culture morphology. A 13-fold increase in final cell count was observed in space with respect to the ground controls and the space flight cells were able to grow in the presence of normally inhibitory levels of gentamicin sulfate. Contrast light microscopy and focused ion beam/scanning electron microscopy showed that, on average, cells in space were 37% of the volume of their matched controls, which may alter the rate of molecule–cell interactions in a diffusion-limited mass transport regime as is expected to occur in microgravity. TEM imagery showed an increase in cell envelope thickness of between 25 and 43% in space with respect to the Earth control group. Outer membrane vesicles were observed on the spaceflight samples, but not on the Earth cultures. While E. coli suspension cultures on Earth were homogenously distributed throughout the liquid medium, in space they tended to form a cluster, leaving the surrounding medium visibly clear of cells. This cell aggregation behavior may be associated with enhanced biofilm formation observed in other spaceflight experiments. PMID:28894439

Eater and draper are involved in the periostial haemocyte immune response in the mosquito Anopheles gambiae.

PubMed

Sigle, L T; Hillyer, J F

2018-03-09

Haemocytes respond to infection by phagocytosing pathogens, producing the enzymes that drive the phenoloxidase-based melanization cascade, secreting lytic factors, and producing other humoral proteins. A subset of haemocytes, called periostial haemocytes, aggregate on the surface of the heart of mosquitoes and kill pathogens in areas of high haemolymph flow. Periostial haemocytes are always present, but an infection induces the recruitment of additional haemocytes to these regions. Here, we tested whether members of the Nimrod gene family are involved in the periostial immune response of the African malaria mosquito, Anopheles gambiae. Using organismal manipulations, RNA interference (RNAi) and microscopy, we show that, following an infection with Escherichia coli, nimrod - the orthologue of Drosophila NimB2 - is not involved in periostial responses. At 4 h postinfection, however, RNAi-based knockdown of draper results in a marginal increase in the number of periostial haemocytes and a doubling of E. coli accumulation at the periostial regions. Finally, at 24 h postinfection, knockdown of eater decreases the number of periostial haemocytes and decreases the phagocytosis of E. coli on the surface of the heart. Phagocytosis of bacteria is more prevalent in the periostial regions of the mid abdominal segments, and knockdown of draper, nimrod or eater does not alter this distribution. Finally, knockdown of Nimrod family genes did not have a meaningful effect on the accumulation of melanin at the periostial regions. This study identifies roles for eater and draper in the functional integration of the mosquito immune and circulatory systems. © 2018 The Royal Entomological Society.

SPECIFIC ADHESION TO CELLULOSE AND HYDROLYSIS OF ORGANOPHOSPHATE NERVE AGENTS BY A GENETICALLY ENGINEERED E. COLI WITH SURFACE-EXPRESSED CELLULOSE-BINDING DOMAIN AND ORGANOPHOSPHORUS HYDROLASE. (R827227)

EPA Science Inventory

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

MICROBIAL BIOSENSOR FOR DIRECT DETERMINATION OF ORGANOPHOSPHATE NERVE AGENTS USING RECOMBINANT ESCHERICHIA COLI WITH SURFACE-EXPRESSED ORGANOPHOSPHORUS HYDROLASE. 2. FIBER-OPTIC MICROBIAL BIOSENSOR. (R823663)

EPA Science Inventory

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

Alternatives for Jet Engine Control. Volume 1: Modelling and Control Design with Jet Engine Data

NASA Technical Reports Server (NTRS)

Sain, M. K.

1985-01-01

This document compiles a comprehensive list of publications supported by, or related to, National Aeronautics and Space Administration Grant NSG-3048, entitled "Alternatives for Jet Engine Control". Dr. Kurt Seldner was the original Technical Officer for the grant, at Lewis Research Center. Dr. Bruce Lehtinen was the final Technical Officer. At the University of Notre Dame, Drs. Michael K. Sain and R. Jeffrey Leake were the original Project Directors, with Dr. Sain becoming the final Project Director. Publications cover work over a ten-year period. The Final Report is divided into two parts. Volume i, "Modelling and Control Design with Jet Engine Data", follows in this report. Volume 2, "Modelling and Control Design with Tensors", has been bound separately.

Systems metabolic engineering of Escherichia coli for L-threonine production.

PubMed

Lee, Kwang Ho; Park, Jin Hwan; Kim, Tae Yong; Kim, Hyun Uk; Lee, Sang Yup

2007-01-01

Amino-acid producers have traditionally been developed by repeated random mutagenesis owing to the difficulty in rationally engineering the complex and highly regulated metabolic network. Here, we report the development of the genetically defined L-threonine overproducing Escherichia coli strain by systems metabolic engineering. Feedback inhibitions of aspartokinase I and III (encoded by thrA and lysC, respectively) and transcriptional attenuation regulations (located in thrL) were removed. Pathways for Thr degradation were removed by deleting tdh and mutating ilvA. The metA and lysA genes were deleted to make more precursors available for Thr biosynthesis. Further target genes to be engineered were identified by transcriptome profiling combined with in silico flux response analysis, and their expression levels were manipulated accordingly. The final engineered E. coli strain was able to produce Thr with a high yield of 0.393 g per gram of glucose, and 82.4 g/l Thr by fed-batch culture. The systems metabolic engineering strategy reported here may be broadly employed for developing genetically defined organisms for the efficient production of various bioproducts.

Identification and Structure–Activity Relationships of Novel Compounds that Potentiate the Activities of Antibiotics in Escherichia coli

DOE PAGES

Haynes, Keith M.; Abdali, Narges; Jhawar, Varsha; ...

2017-06-26

In Gram-negative bacteria, efflux pumps are able to prevent effective cellular concentrations from being achieved for a number of antibiotics. Small molecule adjuvants that act as efflux pump inhibitors (EPIs) have the potential to reinvigorate existing antibiotics that are currently ineffective due to efflux mechanisms. Through a combination of rigorous experimental screening and in silico virtual screening, we recently identified novel classes of EPIs that interact with the membrane fusion protein AcrA, a critical component of the AcrAB-TolC efflux pump in Escherichia coli. In this paper, we present initial optimization efforts and structure–activity relationships around one of those previously describedmore » hits, NSC 60339 (1). Finally, from these efforts we identified two compounds, SLUPP-225 (17h) and SLUPP-417 (17o), which demonstrate favorable properties as potential EPIs in E. coli cells including the ability to penetrate the outer membrane, improved inhibition of efflux relative to 1, and potentiation of the activity of novobiocin and erythromycin.« less

Outbreaks of Illness Associated with Recreational Water--United States, 2011-2012.

PubMed

Hlavsa, Michele C; Roberts, Virginia A; Kahler, Amy M; Hilborn, Elizabeth D; Mecher, Taryn R; Beach, Michael J; Wade, Timothy J; Yoder, Jonathan S

2015-06-26

Outbreaks of illness associated with recreational water use result from exposure to chemicals or infectious pathogens in recreational water venues that are treated (e.g., pools and hot tubs or spas) or untreated (e.g., lakes and oceans). For 2011-2012, the most recent years for which finalized data were available, public health officials from 32 states and Puerto Rico reported 90 recreational water-associated outbreaks to CDC's Waterborne Disease and Outbreak Surveillance System (WBDOSS) via the National Outbreak Reporting System (NORS). The 90 outbreaks resulted in at least 1,788 cases, 95 hospitalizations, and one death. Among 69 (77%) outbreaks associated with treated recreational water, 36 (52%) were caused by Cryptosporidium. Among 21 (23%) outbreaks associated with untreated recreational water, seven (33%) were caused by Escherichia coli (E. coli O157:H7 or E. coli O111). Guidance, such as the Model Aquatic Health Code (MAHC), for preventing and controlling recreational water-associated outbreaks can be optimized when informed by national outbreak and laboratory (e.g., molecular typing of Cryptosporidium) data.

Sequence-Specific Targeting of Bacterial Resistance Genes Increases Antibiotic Efficacy

PubMed Central

Wong, Michael; Daly, Seth M.; Greenberg, David E.; Toprak, Erdal

2016-01-01

The lack of effective and well-tolerated therapies against antibiotic-resistant bacteria is a global public health problem leading to prolonged treatment and increased mortality. To improve the efficacy of existing antibiotic compounds, we introduce a new method for strategically inducing antibiotic hypersensitivity in pathogenic bacteria. Following the systematic verification that the AcrAB-TolC efflux system is one of the major determinants of the intrinsic antibiotic resistance levels in Escherichia coli, we have developed a short antisense oligomer designed to inhibit the expression of acrA and increase antibiotic susceptibility in E. coli. By employing this strategy, we can inhibit E. coli growth using 2- to 40-fold lower antibiotic doses, depending on the antibiotic compound utilized. The sensitizing effect of the antisense oligomer is highly specific to the targeted gene’s sequence, which is conserved in several bacterial genera, and the oligomer does not have any detectable toxicity against human cells. Finally, we demonstrate that antisense oligomers improve the efficacy of antibiotic combinations, allowing the combined use of even antagonistic antibiotic pairs that are typically not favored due to their reduced activities. PMID:27631336

MurD enzymes: some recent developments.

PubMed

Šink, Roman; Barreteau, Hélène; Patin, Delphine; Mengin-Lecreulx, Dominique; Gobec, Stanislav; Blanot, Didier

2013-12-01

The synthesis of the peptide stem of bacterial peptidoglycan involves four enzymes, the Mur ligases (MurC, D, E and F). Among them, MurD is responsible for the ATP-dependent addition of d-glutamic acid to UDP-MurNAc-l-Ala, a reaction which involves acyl-phosphate and tetrahedral intermediates. Like most enzymes of peptidoglycan biosynthesis, MurD constitutes an attractive target for the design and synthesis of new antibacterial agents. Escherichia coli MurD has been the first Mur ligase for which the tridimensional (3D) structure was solved. Thereafter, several co-crystal structures with different ligands or inhibitors were released. In the present review, we will deal with work performed on substrate specificity, reaction mechanism and 3D structure of E. coli MurD. Then, a part of the review will be devoted to recent work on MurD orthologs from species other than E. coli and to cellular organization of Mur ligases and in vivo regulation of the MurD activity. Finally, we will review the different classes of MurD inhibitors that have been designed and assayed to date with the hope of obtaining new antibacterial compounds.

Adherent-invasive Escherichia coli in inflammatory bowel disease.

PubMed

Palmela, Carolina; Chevarin, Caroline; Xu, Zhilu; Torres, Joana; Sevrin, Gwladys; Hirten, Robert; Barnich, Nicolas; Ng, Siew C; Colombel, Jean-Frederic

2018-03-01

Intestinal microbiome dysbiosis has been consistently described in patients with IBD. In the last decades, Escherichia coli , and the adherent-invasive E coli (AIEC) pathotype in particular, has been implicated in the pathogenesis of IBD. Since the discovery of AIEC, two decades ago, progress has been made in unravelling these bacteria characteristics and its interaction with the gut immune system. The mechanisms of adhesion of AIEC to intestinal epithelial cells (via FimH and cell adhesion molecule 6) and its ability to escape autophagy when inside macrophages are reviewed here. We also explore the existing data on the prevalence of AIEC in patients with Crohn's disease and UC, and the association between the presence of AIEC and disease location, activity and postoperative recurrence. Finally, we highlight potential therapeutic strategies targeting AIEC colonisation of gut mucosa, including the use of phage therapy, bacteriocins and antiadhesive molecules. These strategies may open new avenues for the prevention and treatment of IBD in the future. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Assessment of gastrointestinal parasites in wild chimpanzees (Pan troglodytes troglodytes) in southeast Cameroon

PubMed Central

Bertout, Sébastien; Locatelli, Sabrina; Butel, Christelle; Pion, Sébastien; Mpoudi-Ngole, Eitel; Delaporte, Eric; Peeters, Martine; Mallié, Michèle

2014-01-01

We tested 114 faecal samples from wild simian immunodeficiency virus (SIV)-positive (n=43) and SIV-negative (n=71) chimpanzees (Pan troglodytes troglodytes) in southeast Cameroon for the presence of gastrointestinal parasites by direct smear. We observed cysts from different protozoa (Entamoeba coli and Entamoeba histolytica/Entamoeba dispar, Endolimax nana, Iodamoeba butschlii, Chilomastix mesnili, Balantidium coli and Blastocystis cells) and trophozoites from Troglodytella abrassarti and Balantidium coli. Eggs from different helminths (strongylids, Ascaris lumbricoides, Abbreviata caucasica, Trichuris sp., Capillaria sp., Enterobius anthropopeci, Bertiella sp., Hymenolepis diminuta and an undetermined fluke) were also observed. Finally, we observed eggs that could not be properly identified and classified. We did not observe any differences between the SIV+ and SIV− samples except for the unidentified eggs. The studied chimpanzees were highly parasitised by strongylid (85.1 % of prevalence), Troglodytella (43.8 %) and Blastocystis (2.9 %), and the frequency of the other parasites ranged from 0.9 to 8.8 %. These high levels of parasite infections could represent an additional burden in a population where there is a high rate of the SIV virus in circulation. PMID:24781023

Assessment of gastrointestinal parasites in wild chimpanzees (Pan troglodytes troglodytes) in southeast Cameroon.

PubMed

Drakulovski, Pascal; Bertout, Sébastien; Locatelli, Sabrina; Butel, Christelle; Pion, Sébastien; Mpoudi-Ngole, Eitel; Delaporte, Eric; Peeters, Martine; Mallié, Michèle

2014-07-01

We tested 114 faecal samples from wild simian immunodeficiency virus (SIV)-positive (n = 43) and SIV-negative (n = 71) chimpanzees (Pan troglodytes troglodytes) in southeast Cameroon for the presence of gastrointestinal parasites by direct smear. We observed cysts from different protozoa (Entamoeba coli and Entamoeba histolytica / Entamoeba dispar, Endolimax nana, Iodamoeba butschlii, Chilomastix mesnili, Balantidium coli and Blastocystis cells) and trophozoites from Troglodytella abrassarti and Balantidium coli. Eggs from different helminths (strongylids, Ascaris lumbricoides, Abbreviata caucasica, Trichuris sp., Capillaria sp., Enterobius anthropopeci, Bertiella sp., Hymenolepis diminuta and an undetermined fluke) were also observed. Finally, we observed eggs that could not be properly identified and classified. We did not observe any differences between the SIV+ and SIV- samples except for the unidentified eggs. The studied chimpanzees were highly parasitised by strongylid (85.1% of prevalence), Troglodytella (43.8%) and Blastocystis (2.9%), and the frequency of the other parasites ranged from 0.9 to 8.8%. These high levels of parasite infections could represent an additional burden in a population where there is a high rate of the SIV virus in circulation.

Enchancement of Gamma-Aminobutyric Acid Production by Co-Localization of Neurospora crassa OR74A Glutamate Decarboxylase with Escherichia coli GABA Transporter Via Synthetic Scaffold Complex.

PubMed

Somasundaram, Sivachandiran; Maruthamuthu, Murali Kannan; Ganesh, Irisappan; Eom, Gyeong Tae; Hong, Soon Ho

2017-09-28

Gamma-aminobutyric acid is a precursor of nylon-4, which is a promising heat-resistant biopolymer. GABA can be produced from the decarboxylation of glutamate by glutamate decarboxylase. In this study, a synthetic scaffold complex strategy was employed involving the Neurospora crassa glutamate decarboxylase (GadB) and Escherichia coli GABA antiporter (GadC) to improve GABA production. To construct the complex, the SH3 domain was attached to the N. crassa GadB, and the SH3 ligand was attached to the N-terminus, middle, and C-terminus of E. coli GadC. In the C-terminus model, 5.8 g/l of GABA concentration was obtained from 10 g/l glutamate. When a competing pathway engineered strain was used, the final GABA concentration was further increased to 5.94 g/l, which corresponds to 97.5% of GABA yield. With the introduction of the scaffold complex, the GABA productivity increased by 2.9 folds during the initial culture period.

Identification and Structure–Activity Relationships of Novel Compounds that Potentiate the Activities of Antibiotics in Escherichia coli

DOE Office of Scientific and Technical Information (OSTI.GOV)

Haynes, Keith M.; Abdali, Narges; Jhawar, Varsha

In Gram-negative bacteria, efflux pumps are able to prevent effective cellular concentrations from being achieved for a number of antibiotics. Small molecule adjuvants that act as efflux pump inhibitors (EPIs) have the potential to reinvigorate existing antibiotics that are currently ineffective due to efflux mechanisms. Through a combination of rigorous experimental screening and in silico virtual screening, we recently identified novel classes of EPIs that interact with the membrane fusion protein AcrA, a critical component of the AcrAB-TolC efflux pump in Escherichia coli. In this paper, we present initial optimization efforts and structure–activity relationships around one of those previously describedmore » hits, NSC 60339 (1). Finally, from these efforts we identified two compounds, SLUPP-225 (17h) and SLUPP-417 (17o), which demonstrate favorable properties as potential EPIs in E. coli cells including the ability to penetrate the outer membrane, improved inhibition of efflux relative to 1, and potentiation of the activity of novobiocin and erythromycin.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tan, Kemin; Zhou, Qingxuan; Cheng, Bokun

Escherichia coli topoisomerase I has an essential function in preventing hypernegative supercoiling of DNA. A full length structure of E. coli topoisomerase I reported here shows how the C-terminal domains bind single-stranded DNA (ssDNA) to recognize the accumulation of negative supercoils in duplex DNA. These C-terminal domains of E. coli topoisomerase I are known to interact with RNA polymerase, and two flexible linkers within the C-terminal domains may assist in the movement of the ssDNA for the rapid removal of transcription driven negative supercoils. The structure has also unveiled for the first time how the 4-Cys zinc ribbon domain andmore » zinc ribbon-like domain bind ssDNA with primarily π -stacking interactions. Finally, this novel structure, in combination with new biochemical data, provides important insights into the mechanism of genome regulation by type IA topoisomerases that is essential for life, as well as the structures of homologous type IA TOP3α and TOP3β from higher eukaryotes that also have multiple 4-Cys zinc ribbon domains required for their physiological functions.« less

β-galactosidase-catalyzed synthesis of galactosyl chlorphenesin and its characterization.

PubMed

Lee, Sang-Eun; Jo, Tae-Min; Lee, Hyang-Yeol; Lee, Jongsung; Jung, Kyung-Hwan

2013-11-01

We synthesized galactosyl chlorphenesin (CPN-G) using β-gal-containing Escherichia coli (E. coli) cells in which the conversion yield of chlorphenesin (CPN) to CPN-G reached about 64 % during 12 h. CPN-G was identified and characterized using high-performance liquid chromatography, liquid chromatography-mass spectrometry, Fourier transform-infrared spectrometry, and nuclear magnetic resonance analysis ((1)H and (13)C). We verified that a galactose was covalently bound to a CPN alcohol group during CPN-G synthesis throughout these analyses. In particular, by the hydrolysis of CPN-G using β-gal, it was confirmed that a galactose was bound to CPN. The minimal inhibitory concentration (MIC) results showed that the CPN-G MICs were fairly similar to those of CPN. HACAT cell viability was significantly higher in CPN-G-treated cells than in CPN-treated cells at concentrations of 0.0-20.0 mM. Finally, we accomplished the synthesis of less toxic CPN-G, compared with CPN, using β-gal-containing E. coli cells as whole cells without changes in the MICs against microorganisms.

48 CFR 252.235-7011 - Final scientific or technical report.

Code of Federal Regulations, 2014 CFR

2014-10-01

... technical report. 252.235-7011 Section 252.235-7011 Federal Acquisition Regulations System DEFENSE ACQUISITION REGULATIONS SYSTEM, DEPARTMENT OF DEFENSE CLAUSES AND FORMS SOLICITATION PROVISIONS AND CONTRACT... of the report; and (c) For submission of reports in other than paper copy, contact the Defense...

48 CFR 252.235-7011 - Final scientific or technical report.

Code of Federal Regulations, 2012 CFR

2012-10-01

... technical report. 252.235-7011 Section 252.235-7011 Federal Acquisition Regulations System DEFENSE ACQUISITION REGULATIONS SYSTEM, DEPARTMENT OF DEFENSE CLAUSES AND FORMS SOLICITATION PROVISIONS AND CONTRACT... of the report; and (c) For submission of reports in other than paper copy, contact the Defense...

48 CFR 252.235-7011 - Final scientific or technical report.

Code of Federal Regulations, 2010 CFR

2010-10-01

... technical report. 252.235-7011 Section 252.235-7011 Federal Acquisition Regulations System DEFENSE ACQUISITION REGULATIONS SYSTEM, DEPARTMENT OF DEFENSE CLAUSES AND FORMS SOLICITATION PROVISIONS AND CONTRACT... of the report; and (c) For submission of reports in other than paper copy, contact the Defense...

48 CFR 252.235-7011 - Final scientific or technical report.

Code of Federal Regulations, 2011 CFR

2011-10-01

... technical report. 252.235-7011 Section 252.235-7011 Federal Acquisition Regulations System DEFENSE ACQUISITION REGULATIONS SYSTEM, DEPARTMENT OF DEFENSE CLAUSES AND FORMS SOLICITATION PROVISIONS AND CONTRACT... of the report; and (c) For submission of reports in other than paper copy, contact the Defense...

48 CFR 252.235-7011 - Final scientific or technical report.

Code of Federal Regulations, 2013 CFR

2013-10-01

... technical report. 252.235-7011 Section 252.235-7011 Federal Acquisition Regulations System DEFENSE ACQUISITION REGULATIONS SYSTEM, DEPARTMENT OF DEFENSE CLAUSES AND FORMS SOLICITATION PROVISIONS AND CONTRACT... of the report; and (c) For submission of reports in other than paper copy, contact the Defense...

Vocational Outreach Involving Community Exchange (VOICE). Final Report.

ERIC Educational Resources Information Center

Huckabee, Johnni

A Jonesboro, Arkansas project was designed and implemented to increase the awareness of vocational education, increase community support and involvement in vocational education, and establish improved communication in the vocational-technical education field. Interaction between the vocational-technical school and the local school community was a…

King County Metro Transit Hybrid Articulated Buses: Final Evaluation Results

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chandler, K.; Walkowicz, K.

2006-12-01

Final technical report compares and evaluates new diesel and diesel hybrid-electric articulated buses operated as part of the King County Metro Transit (KC Metro) fleet in Seattle, Washington. The evaluation lasted 12 months.

78 FR 1143 - Explosive Siting Requirements; Correction

Federal Register 2010, 2011, 2012, 2013, 2014

2013-01-08

... launch site operators in site planning for the storage and handling of energetic liquids and explosives...: For technical questions concerning this final rule, contact Yvonne Tran, Commercial Space... this final rule, contact Laura Montgomery, AGC 200, [[Page 1144

Final June Revisions Rule State Budgets and New Unit Set-Asides TSD

EPA Pesticide Factsheets

This technical support document (TSD) for the final revisions to the Transport Rule shows the underlying data and calculations used to quantify the state budget revisions and new unit set-aside revisions.

Co-electrospinning of bacteria and viruses

NASA Astrophysics Data System (ADS)

Salalha, Wael; Kuhn, Jonathan; Chervinsky, Shmuel; Zussman, Eyal

2006-03-01

Co-electrospinning provides a novel and highly versatile approach towards composite fibers with diameters ranging from a few hundred nm down to 30 nm with embedded elements. In the present work, co-electrospinning of poly(vinyl alcohol) (PVA) and viruses (T7, T4, λ) or bacteria (Escherichia coli, Staphylococcus albus) was carried out. These preparations should have applications for tissue engineering, gene therapy, phage therapy and biosensing. The average diameter of the co-spun nanofibers was about 300 nm. We found that the encapsulated viruses and bacteria manage to survive the electrospinning process, its pressure buildup in the core of the fiber and the electrostatic field in the co-electrospinning process. Approximately 10% of the Escherichia coli and 20% of Staphylococcus albus cells are viable after spinning. Approximately 5% of the bacterial viruses were also viable after the electrospinning. It should be noted that the encapsulated cells and viruses remain stable for two months without a further decrease in number. These results demonstrate the potential of the co-electrospinning process for the encapsulation and immobilization of bio-objects and the possibility of adapting them to technical applications (e.g., bio-chips).

Department of Defense Access to Intellectual Property for Weapon Systems Sustainment

DTIC Science & Technology

2017-05-01

and acquiring technical data rights …  The cost benefit analysis of including a priced contract option for the future delivery of technical data...entail in terms of cost and benefits , while one of the activities to be finalized is the contract-specific technical data elements.66...Virginia 22311-1882 May 2017 Approved for public release; distribution is unlimited. IDA Paper P-8266 Log: H 17-000030 About This Publication This

10 CFR 51.92 - Supplement to the final environmental impact statement.

Code of Federal Regulations, 2011 CFR

2011-01-01

... changes in the proposed action that are relevant to environmental concerns; or (2) There are new and..., technical, and other benefits and costs of the proposed action, to the extent that the final environmental... costs; (5) Include an analysis of other energy alternatives, to the extent that the final environmental...

Dissemination of Continuing Education Materials Via Television Delivery Systems. Final Technical Report and Final Report.

ERIC Educational Resources Information Center

Munushian, Jack

In 1972, the University of Southern California School of Engineering established a 4-channel interactive instructional television network. It was designed to allow employees of participating industries to take regular university science and engineering courses and special continuing education courses at or near their work locations. Final progress…

The role of non-technical skills in surgery

PubMed Central

Agha, Riaz A.; Fowler, Alexander J.; Sevdalis, Nick

2015-01-01

Non-technical skills are of increasing importance in surgery and surgical training. A traditional focus on technical skills acquisition and competence is no longer enough for the delivery of a modern, safe surgical practice. This review discusses the importance of non-technical skills and the values that underpin successful modern surgical practice. This narrative review used a number of sources including written and online, there was no specific search strategy of defined databases. Modern surgical practice requires; technical and non-technical skills, evidence-based practice, an emphasis on lifelong learning, monitoring of outcomes and a supportive institutional and health service framework. Finally these requirements need to be combined with a number of personal and professional values including integrity, professionalism and compassionate, patient-centred care. PMID:26904193

Analysis of the Bacterial Heat Shock Response to Photodynamic Therapy-Mediated Oxidative Stress

PubMed Central

St. Denis, Tyler G.; Huang, Liyi; Dai, Tianhong; Hamblin, Michael R.

2011-01-01

Antimicrobial photodynamic therapy (PDT) has recently emerged as an effective modality for the selective destruction of bacteria and other pathogenic microorganisms. We investigated whether PDT induced protective responses such as heat shock proteins in bacteria. Using the photosensitizer Toluidine Blue O (TBO) at sub-lethal PDT conditions, a 7-fold increase in bacterial heat shock protein GroEL and a 3-fold increase in heat shock protein DnaK were observed in Escherichia coli post PDT. Pretreatment with 50o C heat for 30 minutes reduced PDT killing in both E. coli and in Enterococcus faecalis, with the most pronounced inhibition occurring at 50-μM TBO with 5-J/cm2 635 nm light, where E. coli killing was reduced by 2- log10 and E. faecalis killing was reduced by 4-log10. Finally, inhibition of the highly conserved chaperone DnaK using a small molecule benzylidene lactam heat shock protein inhibitor potentiated (but not significantly) the effect of PDT at a TBO concentration of 2.5 μM in E. faecalis; however, this effect was not observed in E. coli presumably because inhibitor could not gain access due to Gram-negative permeability barrier. Induction of heat shock proteins may be a mechanism whereby bacteria could become resistant to PDT and warrants the need for further study in the application of dual PDT-heat shock protein-inhibition therapies. PMID:21261628

Semirational Approach for Ultrahigh Poly(3-hydroxybutyrate) Accumulation in Escherichia coli by Combining One-Step Library Construction and High-Throughput Screening.

PubMed

Li, Teng; Ye, Jianwen; Shen, Rui; Zong, Yeqing; Zhao, Xuejin; Lou, Chunbo; Chen, Guo-Qiang

2016-11-18

As a product of a multistep enzymatic reaction, accumulation of poly(3-hydroxybutyrate) (PHB) in Escherichia coli (E. coli) can be achieved by overexpression of the PHB synthesis pathway from a native producer involving three genes phbC, phbA, and phbB. Pathway optimization by adjusting expression levels of the three genes can influence properties of the final product. Here, we reported a semirational approach for highly efficient PHB pathway optimization in E. coli based on a phbCAB operon cloned from the native producer Ralstonia entropha (R. entropha). Rationally designed ribosomal binding site (RBS) libraries with defined strengths for each of the three genes were constructed based on high or low copy number plasmids in a one-pot reaction by an oligo-linker mediated assembly (OLMA) method. Strains with desired properties were evaluated and selected by three different methodologies, including visual selection, high-throughput screening, and detailed in-depth analysis. Applying this approach, strains accumulating 0%-92% PHB contents in cell dry weight (CDW) were achieved. PHB with various weight-average molecular weights (M w ) of 2.7-6.8 × 10 6 were also efficiently produced in relatively high contents. These results suggest that the semirational approach combining library design, construction, and proper screening is an efficient way to optimize PHB and other multienzyme pathways.

Assessing the growth of Escherichia coli O157:H7 and Salmonella in spinach, lettuce, parsley and chard extracts at different storage temperatures.

PubMed

Posada-Izquierdo, G; Del Rosal, S; Valero, A; Zurera, G; Sant'Ana, A S; Alvarenga, V O; Pérez-Rodríguez, F

2016-06-01

The objective of this work was to study the growth potential of Escherichia coli O157:H7 and Salmonella spp. in leafy vegetable extracts at different temperature conditions. Cocktails of five strains of E. coli O157:H7 and of Salmonella enterica were used. Inoculated aqueous vegetable extracts were incubated at 8, 10, 16 and 20°C during 21 days. Microbial growth was monitored using Bioscreen C(®) . In spinach extract, results showed that for E. coli O157:H7 and Salmonella significant differences (P < 0·05) for μabs (maximum absorbance rate) were obtained. For both pathogens, growth in chard was slightly lower. In contrast, iceberg lettuce and parsley showed the lowest values of μabs , below 0·008 h(-1) . The coefficients of variance (CoV) calculated for the different replicates evidenced that at low temperature (8°C) a more variable behaviour of both pathogens is expected (CoV > 180%). This study provides evidence that aqueous extracts from vegetable tissues can result in distinct growth niche producing different response in various types of vegetables. Finally, these results can be used as basis to establish risk rankings of pathogens and leafy vegetable matrices with relation to their potential growth. © 2016 The Society for Applied Microbiology.

75 FR 16345 - Administrative Practices and Procedures; Good Guidance Practices; Technical Amendment

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-01

.... FDA-1999-N-3539] (formerly Docket No. 1999N-4783) Administrative Practices and Procedures; Good Guidance Practices; Technical Amendment AGENCY: Food and Drug Administration, HHS. ACTION: Final rule... Subjects in 21 CFR Part 10 Administrative practice and procedure, News media. 0 Therefore, under the...

Final report : for the period of December 1999 through November 30, 2000 : Florida Transit Training Program (1999/2000) : Florida Technical Assistance Program (1999/2000)

DOT National Transportation Integrated Search

2000-01-01

The following progress report is intended to highlight the significant activities of the Florida Transit Training Program and Florida Technical Assistant Program. The following progress report is intended to highlight the significant activities of th...

78 FR 34264 - Technical Corrections to the HIPAA Privacy, Security, and Enforcement Rules

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-07

...-AA03 Technical Corrections to the HIPAA Privacy, Security, and Enforcement Rules AGENCY: Office for... corrections address certain inadvertent errors and omissions in the HIPAA Privacy, Security, and Enforcement... (HHS or ``the Department'') published a final rule to implement changes to the HIPAA Privacy, Security...

76 FR 11414 - Defense Federal Acquisition Regulation Supplement; Independent Research and Development Technical...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-03-02

... Acquisition Regulation Supplement; Independent Research and Development Technical Descriptions (DFARS Case... (DFARS) to require contractors to report independent research and development (IR&D) projects generating... the address shown below on or before May 2, 2011, to be considered in the formation of the final rule...

Professional Personnel Evaluation System: 1983-84 Final Technical Report.

ERIC Educational Resources Information Center

Doss, David A.

This technical report summarizes the evaluation ratings given to professionals on probation or up for contract renewal in the Austin Independent School District. Graphs, tables, and rankings are presented for each specified population: all district teachers combined; teachers by school level (elementary, junior high, senior high); teachers by…

High School Graduation Minimum Competency Requirements. Final Technical Report.

ERIC Educational Resources Information Center

Austin Independent School District, TX. Office of Research and Evaluation.

This technical report details the testing results and analyses supporting the evaluation findings related to the Austin (Texas) Independent School District (AISD) minimum competency graduation requirements. The graduation competency status of all AISD students in grades 8 to 12 are documented. The report provides additional information on the data…

76 FR 16531 - Technical Correction for Neurological Listing Cross-Reference

Federal Register 2010, 2011, 2012, 2013, 2014

2011-03-24

... 1-800-325-0778, or visit our Internet site, Social Security Online, at http://www.socialsecurity.gov... SOCIAL SECURITY ADMINISTRATION 20 CFR Part 404 [Docket No. SSA-2011-0019] RIN 0960-AH33 Technical Correction for Neurological Listing Cross-Reference AGENCY: Social Security Administration. ACTION: Final...

Improving Vocational Education in Post-High School Institutions. Final Report.

ERIC Educational Resources Information Center

MacArthur, Earl W.

Four ongoing programs in postsecondary vocational-technical education were examined in a national institute attended by 59 representatives from 31 states. Institutions reporting programs were: (1) Los Angeles Trade and Technical College, California, (2) Washtenaw Community College, Ann Arbor, Michigan, (3) Rockingham Community College, Wentworth,…

Chicago Manufacturing Tech Prep. Fiscal Year 1991 Final Report.

ERIC Educational Resources Information Center

Chicago City Colleges, IL.

During its first year of development in 1991, the Chicago Manufacturing Technical Preparation (Tech Prep) Program established a plan for implementing an industry-driven, articulated 4-year manufacturing technology course of study that integrates applied academic courses with technical courses and meets industry hiring standards. The project…

78 FR 29239 - Final Priority; Technical Assistance To Improve State Data Capacity-National Technical Assistance...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-05-20

... reporting requirements. Changes: None. Focus TA on Assessment and Discipline Data Comment: Three commenters agreed with the importance of focusing on assessment and discipline data, and two commenters agreed with... decisions and actions associated with data collection and reporting. One commenter stated that assessment...

Modular Radioisotope Thermoelectric Generator (RTG) Program. Final technical report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

1992-12-31

Section 2.0 of this report summarizes the MOD-RTG reference flight design, and Section 3.0 discusses the Ground Demonstration System design. Multicouple technology development is discussed in Section 4.0, and Section 5.0 lists all published technical papers prepared during the course of the contract.

Methodology in Seeking Stakeholder Perceptions of Effective Technical Oral Presentations: An Exploratory Pilot Study

ERIC Educational Resources Information Center

Bhattacharyya, Ena; Patil, Arun; Sargunan, Rajeswary Appacutty

2010-01-01

Engineering communication studies indicate the importance of oral presentations as an indispensable component of workplace oral communication activities; however, since there is limited literature regarding stakeholder perceptions of effective presentation skills and attributes in technical oral presentations or final year engineering project…

Additional Final Area Designations and Technical Amendment for the 2012 Annual Fine Particle Standard Established in 2012 - Mar 2015

EPA Pesticide Factsheets

EPA is establishing or revising initial area designations and a technical amendment to correct an inadvertent error in the initial designation for one area for the 2012 annual national ambient air quality standards for fine particle pollution.

75 FR 41404 - List of Approved Spent Fuel Storage Casks: NUHOMS®

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-16

.... The NRC is taking this action because the applicant identified that a certain Technical Specification (TS) for Boral characterization was not written precisely. Specifically, the requirements for meeting... changes to the technical specifications. The NRC also published a direct final rule on May 6, 2010 (75 FR...

Research in Automatic Russian-English Scientific and Technical Lexicography. Final Report.

ERIC Educational Resources Information Center

Wayne State Univ., Detroit, MI.

Techniques of reversing English-Russian scientific and technical dictionaries into Russian-English versions through semi-automated compilation are described. Sections on manual and automatic processing discuss pre- and post-editing, the task program, updater (correction of errors and revision by specialist in a given field), the system employed…

National Data Program for the Marine Environment Technical Development Plan. Final Report, Volume Two.

ERIC Educational Resources Information Center

System Development Corp., Santa Monica, CA.

A national data program for the marine environment is recommended. Volume 2 includes: (1) objectives, scope, and methodology; (2) summary of the technical development plan; (3) agency development plans - Great Lakes and coastal development and (4) marine data network development plans. (Author)

Course of Study Project. Final Report. Vocational-Technical Education Departmental Report, Vol. 17, No. 18.

ERIC Educational Resources Information Center

Pennsylvania State Univ., University Park. Div. of Occupational and Vocational Studies.

Designed for use by area vocational-technical schools and other vocational programs, this project developed courses of study in eight occupational areas: commercial art, appliance repair, automotive mechanics, graphic arts, building trades maintenance, building construction trades, diesel mechanics, and welding. Course-of-study development…

Seminar for Preparation of Professional Personnel for Vocational-Technical Education. Final Report.

ERIC Educational Resources Information Center

Dillon, Roy D.; Horner, James T.

Seminar participants included college administrative officers, state vocational education directors, vocational-technical teacher educators, and Office of Education staff. The purpose of the June, 1968 seminar was to consider strategies for resolving critical vocational education personnel supply and demand problems. Presentations included in the…

75 FR 33167 - Technical Amendment Language Change From “Wholly” to “Fully”

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-11

... SOCIAL SECURITY ADMINISTRATION 20 CFR Parts 404, 405, 408, 416, and 418 [Docket No. SSA-2009-0062] RIN 0960-AH16 Technical Amendment Language Change From ``Wholly'' to ``Fully'' AGENCY: Social Security... these final rules, call Brian J. Rudick, Office of Regulations, Social Security Administration, 6401...

76 FR 58144 - Defense Federal Acquisition Regulation Supplement; Presumption of Development Exclusively at...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-09-20

... technical data package, in cases where the Government may have funded only a small portion of the... subcontractor's asserted restrictions on technical data and computer software. DATES: Effective date: September... data and computer software. More specifically, the final rule affects these validation procedures in...

75 FR 49408 - Navigation and Navigable Waters; Technical, Organizational, and Conforming Amendments, Bridges

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-13

... Conforming Amendments, Bridges AGENCY: Coast Guard, DHS. ACTION: Final rule. SUMMARY: This rule makes non... technical corrections to Coast Guard bridge and navigable waters regulations. This rule will have no... announces or gathers public opinion or other information regarding bridge matters, nor will it change the...

Research in network management techniques for tactical data communications networks

NASA Astrophysics Data System (ADS)

Boorstyn, R.; Kershenbaum, A.; Maglaris, B.; Sarachik, P.

1982-09-01

This is the final technical report for work performed on network management techniques for tactical data networks. It includes all technical papers that have been published during the control period. Research areas include Packet Network modelling, adaptive network routing, network design algorithms, network design techniques, and local area networks.

A System for Evaluating Microcomputer Courseware for Vocational and Technical Education. Final Report.

ERIC Educational Resources Information Center

Chase, Shirley A.; And Others

A project was conducted to design a system for evaluating microcomputer courseware for vocational and technical education. Through a literature review and contacts with organizations and individuals involved in courseware evaluation and use, project staff identified and acquired for review documents pertaining to courseware evaulation, vocational…

Simultaneous detection of Escherichia coli O157:H7 and Salmonella Typhimurium using quantum dots as fluorescence labels.

PubMed

Yang, Liju; Li, Yanbin

2006-03-01

In this study, we explored the use of semiconductor quantum dots (QDs) as fluorescence labels in immunoassays for simultaneous detection of two species of foodborne pathogenic bacteria, Escherichia coli O157:H7 and Salmonella Typhimurium. QDs with different sizes can be excited with a single wavelength of light, resulting in different emission peaks that can be measured simultaneously. Highly fluorescent semiconductor quantum dots with different emission wavelengths (525 nm and 705 nm) were conjugated to anti-E. coli O157 and anti-Salmonella antibodies, respectively. Target bacteria were separated from samples by using specific antibody coated magnetic beads. The bead-cell complexes reacted with QD-antibody conjugates to form bead-cell-QD complexes. Fluorescent microscopic images of QD labeled E. coli and Salmonella cells demonstrated that QD-antibody conjugates could evenly and completely attach to the surface of bacterial cells, indicating that the conjugated QD molecules still retain their effective fluorescence, while the conjugated antibody molecules remain active and are able to recognize their specific target bacteria in a complex mixture. The intensities of fluorescence emission peaks at 525 nm and 705 nm of the final complexes were measured for quantitative detection of E. coli O157:H7 and S. Typhimurium simultaneously. The fluorescence intensity (FI) as a function of cell number (N) was found for Salmonella and E. coli, respectively. The regression models can be expressed as: FI = 60.6 log N- 250.9 with R(2) = 0.97 for S. Typhimurium, and FI = 77.8 log N- 245.2 with R(2) = 0.91 for E. coli O157:H7 in the range of cell numbers from 10(4) to 10(7) cfu ml(-1). The detection limit of this method was 10(4) cfu ml(-1). The detection could be completed within 2 hours. The principle of this method could be extended to detect multiple species of bacteria (3-4 species) simultaneously, depending on the availability of each type of QD-antibody conjugates with a unique emission peak and the antibody coated magnetic beads specific to each species of bacteria.

An electrochemical immunoassay for Escherichia coli O157:H7 using double functionalized Au@Pt/SiO2 nanocomposites and immune magnetic nanoparticles.

PubMed

Ye, Lingxian; Zhao, Guangying; Dou, Wenchao

2018-05-15

A sensitive Point-of-Care Testing (POCT) with Au-Pt bimetallic nanoparticles (Au@Pt) functionalized silica nanoparticle (SiO 2 NPs) and Fe 3 O 4 magnetic nanoparticles (Fe 3 O 4 NPs) was designed for the quantitative detection of Escherichia coli O157:H7 (E. coli O157:H7). The poly-(4-styrenesulfonic acid-co-maleic acid) (PSSMA) as a negatively charged polyelectrolyte can be easily coated on surface of the amino group modified SiO 2 NPs via electrostatic force. PSSMA is also a good stabilizer for water-soluble bimetallic nanostructures. The PSSMA is first time used as a "bridge" to connect the negative charge Au@Pt NPs to the SiO 2 NPs, forming Au@Pt/SiO 2 NPs. Antibody and invertase conjugated Au@Pt/SiO 2 NPs (denoted as Ab/invertase-Au@Pt/SiO 2 NPs) were used as signal labels. Monoclonal antibody against E. coli O157:H7 (Ab) functionalized magnetic nanoparticles (denoted as Ab-Fe 3 O 4 @SiO 2 NPs) were used to enrich and capture the E. coli O157:H7 in positive sample. The immunosensing platform also composed of a personal glucometer (PGM) using for signal readout. Based on this sandwich-type immunoassay, the invertase in the final formed sandwich immunocomplex catalyzed the hydrolysis of sucrose to produce a large amount of glucose for quantitative readout by the PGM. Under optimal conditions, a linear relationship between the glucose concentration and the logarithm of E. coli O157:H7 concentration was obtained in the concentration range from 3.5 × 10 2 to 3.5 × 10 8 CFU mL -1 with a detection limit of 1.83 × 10 2 CFU mL -1 (3σ). This method was used to detect E. coli O157:H7 in spiked milk samples, indicating its potential practical application. This protocol can be applied in various fields of study. Copyright © 2018 Elsevier B.V. All rights reserved.

“Direct cloning in Lactobacillus plantarum: Electroporation with non-methylated plasmid DNA enhances transformation efficiency and makes shuttle vectors obsolete”

PubMed Central

2012-01-01

Background Lactic acid bacteria (LAB) play an important role in agricultural as well as industrial biotechnology. Development of improved LAB strains using e.g. library approaches is often limited by low transformation efficiencies wherefore one reason could be differences in the DNA methylation patterns between the Escherichia coli intermediate host for plasmid amplification and the final LAB host. In the present study, we examined the influence of DNA methylation on transformation efficiency in LAB and developed a direct cloning approach for Lactobacillus plantarum CD033. Therefore, we propagated plasmid pCD256 in E. coli strains with different dam/dcm-methylation properties. The obtained plasmid DNA was purified and transformed into three different L. plantarum strains and a selection of other LAB species. Results Best transformation efficiencies were obtained using the strain L. plantarum CD033 and non-methylated plasmid DNA. Thereby we achieved transformation efficiencies of ~ 109 colony forming units/μg DNA in L. plantarum CD033 which is in the range of transformation efficiencies reached with E. coli. Based on these results, we directly transformed recombinant expression vectors received from PCR/ligation reactions into L. plantarum CD033, omitting plasmid amplification in E. coli. Also this approach was successful and yielded a sufficient number of recombinant clones. Conclusions Transformation efficiency of L. plantarum CD033 was drastically increased when non-methylated plasmid DNA was used, providing the possibility to generate expression libraries in this organism. A direct cloning approach, whereby ligated PCR-products where successfully transformed directly into L. plantarum CD033, obviates the construction of shuttle vectors containing E. coli-specific sequences, as e.g. a ColEI origin of replication, and makes amplification of these vectors in E. coli obsolete. Thus, plasmid constructs become much smaller and occasional structural instability or mutagenesis during E. coli propagation is excluded. The results of our study provide new genetic tools for L. plantarum which will allow fast, forward and systems based genetic engineering of this species. PMID:23098256

Survival of artificially inoculated Escherichia coli and Salmonella Typhimurium on the surface of raw poultry products subjected to crust freezing.

PubMed

Chaves, B D; Han, I Y; Dawson, P L; Northcutt, J K

2011-12-01

Escherichia coli and Salmonella spp. are ubiquitous in the poultry production environment, and hence, their transmission to poultry products is of concern. Industry has widely used freezing as a strategy to halt pathogen growth, and more recently, crust freezing has been suggested as a means to improve mechanical operations, quality, and safety of poultry products. The purpose of this study was to evaluate the effect of crust freezing on the survival of Escherichia coli and Salmonella Typhimurium that were artificially inoculated on the surface of raw poultry products with or without adhering skin. Ampicillin-resistant (AR) E. coli JM 109 and nalidixic acid-resistant (NAR) Salmonella Typhimurium were used in the experiments. A set of cultures was subjected to cold-shock stress by storage at 4°C for 10 d. After being either cold-shocked or non-cold-shocked, commercial chicken breasts without skin and chicken thighs with skin were inoculated in separate experiments with each bacterium. Samples were crust frozen at -85°C for 20 min or completely frozen at -85°C for 60 min. The E. coli and Salmonella Typhimurium were recovered on appropriate selective and nonselective media containing the corresponding antibiotic. Log reductions and extent of injury were calculated and treatments were compared using ANOVA. No significant differences were observed in the reduction of cold-shocked or non-cold-shocked bacteria on products with or without skin that were crust or completely frozen. The average reduction for E. coli was 0.15 log(10) cfu/mL of rinse, and for Salmonella Typhimurium 0.10 log(10) cfu/mL of rinse; therefore, none of the final reductions were greater than the desired target (1 log). Bacterial cell injury was not significantly different (P > 0.05) among any of the treatments. Data showed no practical significance for initial reduction of these pathogens from crust freezing and thus, this technology should not be considered as a strategy for the reduction of E. coli and Salmonella Typhimurium on poultry.

Inactivation of Nonpathogenic Escherichia coli, Escherichia coli O157:H7, Salmonella enterica Typhimurium, and Listeria monocytogenes in Ice Using a UVC Light-Emitting Diode.

PubMed

Murashita, Suguru; Kawamura, Shuso; Koseki, Shigenobu

2017-07-01

Ice, widely used in the food industry, is a potential cause of food poisoning resulting from microbial contamination. Direct microbial inactivation of ice is necessary because microorganisms may have been present in the source water used to make it and/or may have been introduced due to poor hygiene during production or handling of the ice. Nonthermal and nondestructive microbial inactivation technologies are needed to control microorganisms in ice. We evaluated the applicability of a UVC light-emitting diode (UVC-LED) for microbial inactivation in ice. The effects of UV intensity and UV dose of the UVC-LED on Escherichia coli ATCC 25922 and a comparison of UVC-LED with a conventional UV lamp for effective bacterial inactivation in distilled water and ice cubes were investigated to evaluate the performance of the UVC-LED. Finally, we assessed the effects of the UVC-LED on pathogens such as E. coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in ice cubes. The results indicated that UVC-LED effectiveness depended on the UV dose at all UV intensity conditions (0.084, 0.025, 0.013, 0.007, and 0.005 mW/cm 2 ) in ice and that UVC-LED could more efficiently inactivate E. coli ATCC 25922 in distilled water and ice than the UV lamp. At a UV dose of 2.64 mJ/cm 2 , E. coli in distilled water was decreased by 0.90 log CFU/mL (UV lamp) and by more than 7.0 log CFU/mL (UVC-LED). At 15.2 mJ/cm 2 , E. coli in ice was decreased by 3.18 log CFU/mL (UV lamp) and by 4.45 CFU/mL (UVC-LED). Furthermore, UVC-LED irradiation reduced the viable number of pathogens by 6 to 7 log cycles at 160 mJ/cm 2 , although the bactericidal effect was somewhat dependent on the type of bacteria. L. monocytogenes in ice was relatively more sensitive to UVC irradiation than were E. coli O157:H7 and Salmonella Typhimurium. These results demonstrate that UVC-LED irradiation could contribute to the safety of ice in the food industry.

Funcationalized nanomaterials and their biological applications

NASA Astrophysics Data System (ADS)

Vedantam, Pallavi

Bionanomaterials have been used in drug delivery, cancer therapy and biodiagnosis of pathogens based on their size and surface functionalization. In this present work, different kinds of nanoparticles (NPs), their cellular interactions, cytotoxicity profiles, and finally role of gold nanoparticles (GNPs) in biodetection of E. coli was investigated. Firstly, cytotoxicity profiles of commercial, laser ablation, and green synthetic NPs were studied. Induction of apoptosis in cancer cells was found to be size dependent. The plain 80 nm GNPs and AgNPs enhanced toxic effects in cancer cells when compared to 20 nm ones. Apoptotic profiles of ALB- or FBS-coated NPs were significantly low in cancer cells when compared to plain NPs. The FBS-coated NPs were relatively bigger. Whereas, green synthetic NPs prepared from floral extracts of Tacoma stans and Tagetus erecta caused significant increase in cell membrane damage in cancer cells when compared to commercial GNPs. The phytochemicals in the extract were found to have a synergistic effect on pathogens like E. coli, Staphylococcus aureus and Enterococcus faecalis when used with antibiotics like tetracycline, ampicillin and vancomycin. This effect can be capitalized in developing NPs as effective drug carriers. Next, use of commercial GNPs as diagnostic tools to detect competitive binding in DU-145 in the presence of UTI-causing E. coli ORN178 was studied. GNPs functionalized with D-mannose (Mn) showed competitive binding between Mn-GNPs and E. coli ORN178 when presented together with DU-145 cells. Cytotoxicity assays of plain and Mn-GNPs showed significant decrease in viability of DU-145 cells. The plain/Mn 20 nm GNPs were taken up more by the cell when compared to the 200 nm ones. The protein-coated GNPs were found to be stable in culture medium. This competitive binding can be further developed to prevent/detect recurrent UTI in DU-45 cells. Lastly, primary and fine sugar specificity of fimbrial lectins of E.coli ORN178 and E. coli 13762 to D-mannose and Neualphac(alpha2-3)-Gal-(beta1-4)Glc-Paa functionalized GNPs showed that E. coli ORN178 binds specifically only to Mn-GNPs and E. coli 13762 to the latter. Hence, adhesin-specific adhesion shows great potential for designing NPs to specifically bind to microorganisms as biodiagnostic tools.

Evaluating the U.S. Food Safety Modernization Act Produce Safety Rule Standard for Microbial Quality of Agricultural Water for Growing Produce.

PubMed

Havelaar, Arie H; Vazquez, Kathleen M; Topalcengiz, Zeynal; Muñoz-Carpena, Rafael; Danyluk, Michelle D

2017-10-09

The U.S. Food and Drug Administration (FDA) has defined standards for the microbial quality of agricultural surface water used for irrigation. According to the FDA produce safety rule (PSR), a microbial water quality profile requires analysis of a minimum of 20 samples for Escherichia coli over 2 to 4 years. The geometric mean (GM) level of E. coli should not exceed 126 CFU/100 mL, and the statistical threshold value (STV) should not exceed 410 CFU/100 mL. The water quality profile should be updated by analysis of a minimum of five samples per year. We used an extensive set of data on levels of E. coli and other fecal indicator organisms, the presence or absence of Salmonella, and physicochemical parameters in six agricultural irrigation ponds in West Central Florida to evaluate the empirical and theoretical basis of this PSR. We found highly variable log-transformed E. coli levels, with standard deviations exceeding those assumed in the PSR by up to threefold. Lognormal distributions provided an acceptable fit to the data in most cases but may underestimate extreme levels. Replacing censored data with the detection limit of the microbial tests underestimated the true variability, leading to biased estimates of GM and STV. Maximum likelihood estimation using truncated lognormal distributions is recommended. Twenty samples are not sufficient to characterize the bacteriological quality of irrigation ponds, and a rolling data set of five samples per year used to update GM and STV values results in highly uncertain results and delays in detecting a shift in water quality. In these ponds, E. coli was an adequate predictor of the presence of Salmonella in 150-mL samples, and turbidity was a second significant variable. The variability in levels of E. coli in agricultural water was higher than that anticipated when the PSR was finalized, and more detailed information based on mechanistic modeling is necessary to develop targeted risk management strategies.

Development of a hierarchical model for predicting microbiological contamination of private groundwater supplies in a geologically heterogeneous region.

PubMed

O'Dwyer, Jean; Hynds, Paul D; Byrne, Kenneth A; Ryan, Michael P; Adley, Catherine C

2018-06-01

Private groundwater sources in the Republic of Ireland provide drinking water to an estimated 750,000 people or 16% of the national population. Consumers of untreated groundwater are at increased risk of infection from pathogenic microorganisms. However, given the volume of private wells in operation, remediation or even quantification of public risk is both costly and time consuming. In this study, a hierarchical logistic regression model was developed to 'predict' contamination with E. coli based on the results of groundwater quality analyses of private wells (n = 132) during the period of September 2011 to November 2012. Assessment of potential microbial contamination risk factors were categorised into three groups: Intrinsic (environmental factors), Specific (local features) and Infrastructural (groundwater source characteristics) which included a total of 15 variables. Overall, 51.4% of wells tested positive for E. coli during the study period with univariate analysis indicating that 11 of the 15 assessed risk factors, including local bedrock type, local subsoil type, septic tank reliance, 5 day antecedent precipitation and temperature, along with well type and depth, were all significantly associated with E. coli presence (p < 0.05). Hierarchical logistic regression was used to develop a private well susceptibility model with the final model containing 8 of the 11 associated variables. The model was shown to be highly efficient; correctly classifying the presence of E. coli in 94.2% of cases, and the absence of E. coli in 84.7% of cases. Model validation was performed using an external data set (n = 32) and it was shown that the model has promising accuracy with 90% of positive E. coli cases correctly predicted. The developed model represents a risk assessment and management tool that may be used to develop effective water-quality management strategies to minimize public health risks both in Ireland and abroad. Copyright © 2018 Elsevier Ltd. All rights reserved.

Final Technical Report, Wind Generator Project (Ann Arbor)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Geisler, Nathan

A Final Technical Report (57 pages) describing educational exhibits and devices focused on wind energy, and related outreach activities and programs. Project partnership includes the City of Ann Arbor, MI and the Ann Arbor Hands-on Museum, along with additional sub-recipients, and U.S. Department of Energy/Office of Energy Efficiency and Renewable Energy (EERE). Report relays key milestones and sub-tasks as well as numerous graphics and images of five (5) transportable wind energy demonstration devices and five (5) wind energy exhibits designed and constructed between 2014 and 2016 for transport and use by the Ann Arbor Hands-on Museum.

Final Technical Report- Virginia Solar Pathways Project

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bond, Katharine; Cosby, Sarah

This Report provides a technical review of the final results of a funding award to Virginia Electric and Power Company (Dominion Energy Virginia (DEV) or the Company) for a project under the U.S. Department of Energy’s Solar Energy Technologies Office. The three-year project was formally known as the Virginia Solar Pathways Project (VSPP or the Project). The purpose of the VSPP was to develop a collaborative utility-administered solar strategy (Solar Strategy) for DEV’s service territory in the Commonwealth that could serve as a replicable model for other states with similar policy environments.

M-X Environmental Technical Report. Socioeconomic Impact Estimates for White Pine County, Nevada. Detailed Tables.

DTIC Science & Technology

1980-12-22

8217AP0A095 772 HENNINGSON DURHAM AND RICHARDSON SANTA BARBARA CA F/G 16/1 B-B ENVIRONMENTAL TECHNICAL REPORT. SOCIOECONOMIC IMPACT ESTIMA--ETC(U) DEC... Environmental Technical Reports,. 53ocioeconctnic’ Final r1eport Impact . Estimates for White Pine County, Nevada. ____________ Detailed Tables 6. PERFORMING...background information for the analysis contained in the M-X Deploment Area Selection and Land Withirawal/Acquisition Draft Environmental Impact Statement

Optical fiber sensors for materials and structures characterization

NASA Technical Reports Server (NTRS)

Lindner, D. K.; Claus, R. O.

1991-01-01

The final technical report on Optical Fiber Sensors for Materials and Structures Characterization, covering the period August 1990 through August 1991 is presented. Research programs in the following technical areas are described; sapphire optical fiber sensors; vibration analysis using two-mode elliptical core fibers and sensors; extrinsic Fabry-Perot interferometer development; and coatings for fluorescent-based sensor. Research progress in each of these areas was substantial, as evidenced by the technical publications which are included as appendices.

A joint university-government technical editing program

NASA Technical Reports Server (NTRS)

Stohrer, Freda F.; Pinelli, Thomas E.

1978-01-01

The NASA Langley Research Center (LaRC) and Old Dominion University have designed a mutually useful technical editing program. A university team made up of an English instructor and two graduate students - one from English, one from engineering - works with a senior editor from LaRC to prepare technical reports for publication. A round-robin technique gives the university team editorial commentary from both language and technical specialists; the senior editor from LaRC supervises reports through final publication. To date, the system has provided LaRC with a respectable product and university students with valuable on-the-job training.

Optimization of an enhanced ceramic micro-filter for concentrating E.coli in water

NASA Astrophysics Data System (ADS)

Zhang, Yushan; Guo, Tianyi; Xu, Changqing; Hong, Lingcheng

2017-02-01

Recently lower limit of detection (LOD) is necessary for rapid bacteria detection and analysis applications in clinical practices and daily life. A critical pre-conditioning step for these applications is bacterial concentration, especially for low level of pathogens. Sample volume can be largely reduced with an efficient pre-concentration process. Some approaches such as hollow-fiber ultra-filtration and electrokinetic technique have been applied to bacterial concentration. Since none of these methods can provide a concentrating method with a stable recovery efficiency, bacterial concentration still remains challenging Ceramic micro- filter can be used to concentrate the bacteria but the cross flow system keeps the bacteria in suspension. Similar harvesting bacteria using ultra-filtration showed an average recovery efficiency of 43% [1] and other studies achieved recovery rates greater than 50% [2]. In this study, an enhanced ceramic micro-filter with 0.14 μm pore size was proposed and demonstrated to optimize the concentration of E.coli. A high recovery rate (mean value >90%) and a high volumetric concentration ratio (>100) were achieved. Known quantities (104 to 106 CFU/ml) of E.coli cells were spiked to different amounts of phosphate buffered saline (0.1 to 1 L), and then concentrated to a final retentate of 5 ml to 10 ml. An average recovery efficiency of 95.3% with a standard deviation of 5.6% was achieved when the volumetric con- centration ratio was 10. No significant recovery rate loss was indicated when the volumetric concentration ratio reached up to 100. The effects of multiple parameters on E.coli recovery rate were also studied. The obtained results indicated that the optimized ceramic micro- filtration system can successfully concentrate E.coli cells in water with an average recovery rate of 90.8%.

Efficacy of adding detergents to sanitizer solutions for inactivation of Escherichia coli O157:H7 on Romaine lettuce.

PubMed

Keskinen, Lindsey A; Annous, Bassam A

2011-06-30

Numerous Escherichia coli O157:H7 outbreaks have been linked to consumption of fresh lettuce. The development of effective and easily implemented wash treatment could reduce such incidents. The purpose of this study was to evaluate the addition of food-grade detergents to sanitizer solutions for inactivation of E. coli O157:H7 on Romaine lettuce. Freshly-cut leaves of Romaine lettuce were dip-inoculated to achieve a final cell concentration of 7.8±0.2 log CFU/g, air-dried for 2h, and stored overnight at 4 °C. Leaves were then washed for 2 min in an experimental short chain fatty acid formulation (SCFA) or in one of the following solutions with or without 0.2% dodecylbenzenesulfonic acid or 0.2% sodium 2-ethyl hexyl sulfate: 1) deionized water; 2) 100 ppm chlorine dioxide; 3) 100 ppm chlorine; and 4) 200 ppm chlorine. Following wash treatment, samples were blended in neutralizing buffer (1:3) and surface plated on the selective media CT-SMAC. The efficacy of wash treatments, with or without the detergents, in inactivating E. coli O157:H7 cells on lettuce leaves were not significantly different. The most effective wash solution was SCFA, which was capable of reducing E. coli O157:H7 populations by more than 5 log CFU/g. The rest of the wash treatments resulted in a population reduction of less than 1 log CFU/g. The effectiveness of SCFA surpasses that of other sanitizer treatments tested in this study and requires further research to optimize treatments to preserve lettuce quality. Conventional detergents did not enhance the efficacy of any of the wash treatments tested during this study. Published by Elsevier B.V.

Survival and growth of Listeria monocytogenes and enterohemorrhagic Escherichia coli O157:H7 in minimally processed artichokes.

PubMed

Sanz, Susana; Giménez, Mercedes; Olarte, Carmen

2003-12-01

The ability of Listeria monocytogenes and Escherichia coli O157:H7 inoculated by immersion (at 4.6 and 5.5 log CFU/ g, respectively) to survive on artichokes during various stages of preparation was determined. Peeling, cutting, and disinfecting operations (immersion in 50 ppm of a free chlorine solution at 4 degrees C for 5 min) reduced populations of L. monocytogenes and E. coli O157:H7 by only 1.6 and 0.8 log units, respectively. An organic acid rinse (0.02% citric acid and 0.2% ascorbic acid) was more effective than a tap water rinse in removing these pathogens. Given the possibility of both pathogens being present on artichokes at the packaging stage, their behavior during the storage of minimally processed artichokes was investigated. For this purpose, batches of artichokes inoculated with L. monocytogenes or E. coli O157:H7 (at 5.5 and 5.2 log CFU/g, respectively) were packaged in P-Plus film bags and stored at 4 degrees C for 16 days. During this period, the equilibrium atmosphere composition and natural background microflora (mesophiles, psychrotrophs, anaerobes, and fecal coliforms) were also analyzed. For the two studied pathogens, the inoculum did not have any effect on the final atmospheric composition (10% O2, 13% CO2) or on the survival of the natural background microflora of the artichokes. L. monocytogenes was able to survive during the entire storage period in the inoculated batches, while the E. coli O157:H7 level increased by 1.5 log units in the inoculated batch during the storage period. The modified atmosphere was unable to control the behavior of either pathogen.

A sensitive biosensor using double-layer capillary based immunomagnetic separation and invertase-nanocluster based signal amplification for rapid detection of foodborne pathogen.

PubMed

Huang, Fengchun; Zhang, Huilin; Wang, Lei; Lai, Weihua; Lin, Jianhan

2018-02-15

Combining double-layer capillary based high gradient immunomagnetic separation, invertase-nanocluster based signal amplification and glucose meter based signal detection, a novel biosensor was developed for sensitive and rapid detection of E. coli O157:H7 in this study. The streptavidin modified magnetic nanobeads (MNBs) were conjugated with the biotinylated polyclonal antibodies against E. coli O157:H7 to form the immune MNBs, which were captured by the high gradient magnetic field in the double-layer capillary to specifically separate and efficiently concentrate the target bacteria. Calcium chloride was used with the monoclonal antibodies against E. coli O157:H7 and the invertase to form the immune invertase-nanoclusters (INCs), which were used to react with the target bacteria to form the MNB-bacteria-INC complexes in the capillary. The sucrose was then injected into the capillary and catalyzed by the invertase on the complexes into the glucose, which was detected using the glucose meter to obtain the concentration of the glucose for final determination of the E. coli O157:H7 cells in the sample. A linear relationship between the readout of the glucose meter and the concentration of the E. coli O157:H7 cells (from 10 2 to 10 7 CFU/mL) was found and the lower detection limit of this biosensor was 79 CFU/mL. This biosensor might be extended for the detection of other foodborne pathogens by changing the antibodies and has shown the potential for the detection of foodborne pathogens in a large volume of sample to further increase the sensitivity. Copyright © 2017 Elsevier B.V. All rights reserved.

Characteristics of Integrons and Associated Gene Cassettes in Antibiotic-Resistant Escherichia coli Isolated from Free-Ranging Food Animals in China.

PubMed

Rehman, Mujeeb Ur; Zhang, Hui; Huang, Shucheng; Iqbal, Muhammad Kashif; Mehmood, Khalid; Luo, Houqiang; Li, Jiakui

2017-08-01

We investigated the occurrence of integrons in antibiotic-resistant Escherichia coli strains isolated from free-ranging food animals, including yaks, piglets, and chickens, in China, and characterized the gene cassettes harbored within the integrons. We examined 432 E. coli strains that exhibited resistance to at least one class of antibiotics. Integrase genes and associated gene cassettes were characterized by polymerase chain reaction (PCR) analysis, restriction fragment-length polymorphism, DNA sequencing, conjugation experiments, and plasmid analysis. Twenty-nine (6.7%) integrons were amplified from the 432 antimicrobial-resistant (AMR) isolates evaluated. Specifically, class 1 and 2 integrons were detected in 26 (6%) and 3 (0.7%) strains, respectively. Meanwhile, 6 different gene cassettes, dfrA1, dfr12, aadA1, aadA2, sat1, and orfF, were detected within 6 variable regions (VRs), of which the dfrA1 + aadA1 array was the most common, identified in 12 of 26 class 1 integrons (46.1%). Meanwhile, only one class 2 integron contained a cassette, and the remaining two contained undetermined VRs. Finally, a conjugation assay confirmed the transfer of 4 different types of class 1 integrons into recipient strains, with plasmid sizes ranging from 20 to 30 kb. This is the first report examining the baseline AMR characteristics of E. coli within an extensive farming system of livestock animals in China. Given that integrons were detected in >6% of resistant E. coli strains, precautionary measures are required to prevent the spread of mobile genetic resistance determinants in food animals and monitor their emergence. © 2017 Institute of Food Technologists®.

The morphogenetic MreBCD proteins of Escherichia coli form an essential membrane-bound complex.

PubMed

Kruse, Thomas; Bork-Jensen, Jette; Gerdes, Kenn

2005-01-01

MreB proteins of Escherichia coli, Bacillus subtilis and Caulobacter crescentus form actin-like cables lying beneath the cell surface. The cables are required to guide longitudinal cell wall synthesis and their absence leads to merodiploid spherical and inflated cells prone to cell lysis. In B. subtilis and C. crescentus, the mreB gene is essential. However, in E. coli, mreB was inferred not to be essential. Using a tight, conditional gene depletion system, we systematically investigated whether the E. coli mreBCD-encoded components were essential. We found that cells depleted of mreBCD became spherical, enlarged and finally lysed. Depletion of each mre gene separately conferred similar gross changes in cell morphology and viability. Thus, the three proteins encoded by mreBCD are all essential and function in the same morphogenetic pathway. Interestingly, the presence of a multicopy plasmid carrying the ftsQAZ genes suppressed the lethality of deletions in the mre operon. Using GFP and cell fractionation methods, we showed that the MreC and MreD proteins were associated with the cell membrane. Using a bacterial two-hybrid system, we found that MreC interacted with both MreB and MreD. In contrast, MreB and MreD did not interact in this assay. Thus, we conclude that the E. coli MreBCD form an essential membrane-bound complex. Curiously, MreB did not form cables in cell depleted for MreC, MreD or RodA, indicating a mutual interdependency between MreB filament morphology and cell shape. Based on these and other observations we propose a model in which the membrane-associated MreBCD complex directs longitudinal cell wall synthesis in a process essential to maintain cell morphology.

OptSSeq: High-throughput sequencing readout of growth enrichment defines optimal gene expression elements for homoethanologenesis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ghosh, Indro Neil; Landick, Robert

The optimization of synthetic pathways is a central challenge in metabolic engineering. OptSSeq (Optimization by Selection and Sequencing) is one approach to this challenge. OptSSeq couples selection of optimal enzyme expression levels linked to cell growth rate with high-throughput sequencing to track enrichment of gene expression elements (promoters and ribosomebinding sites) from a combinatorial library. OptSSeq yields information on both optimal and suboptimal enzyme levels, and helps identify constraints that limit maximal product formation. Here we report a proof-of-concept implementation of OptSSeq using homoethanologenesis, a two-step pathway consisting of pyruvate decarboxylase (Pdc) and alcohol dehydrogenase (Adh) that converts pyruvate tomore » ethanol and is naturally optimized in the bacterium Zymomonas mobilis. We used OptSSeq to determine optimal gene expression elements and enzyme levels for Z. mobilis Pdc, AdhA, and AdhB expressed in Escherichia coli. By varying both expression signals and gene order, we identified an optimal solution using only Pdc and AdhB. We resolved current uncertainty about the functions of the Fe 2+-dependent AdhB and Zn 2+- dependent AdhA by showing that AdhB is preferred over AdhA for rapid growth in both E. coli and Z. mobilis. Finally, by comparing predictions of growth-linked metabolic flux to enzyme synthesis costs, we established that optimal E. coli homoethanologenesis was achieved by our best pdc-adhB expression cassette and that the remaining constraints lie in the E. coli metabolic network or inefficient Pdc or AdhB function in E. coli. Furthermore, OptSSeq is a general tool for synthetic biology to tune enzyme levels in any pathway whose optimal function can be linked to cell growth or survival.« less

Cell surface acid-base properties of Escherichia coli and Bacillus brevis and variation as a function of growth phase, nitrogen source and C:N ratio.

PubMed

Hong, Yongsuk; Brown, Derick G

2006-07-01

Potentiometric titration has been conducted to systematically examine the acid-base properties of the cell surfaces of Escherichia coli K-12 and Bacillus brevis as a function of growth phase, nitrogen source (ammonium or nitrate), and carbon to nitrogen (C:N) ratio of the growth substrate. The two bacterial species revealed four distinct proton binding sites, with pK(a) values in the range of 3.08-4.05 (pK(1)), 4.62-5.57 (pK(2)), 6.47-7.30 (pK(3)), and 9.68-10.89 (pK(4)) corresponding to phosphoric/carboxylic, carboxylic, phosphoric, and hydroxyl/amine groups, respectively. Two general observations in the data are that for B. brevis the first site concentration (N(1)), corresponding to phosphoric/carboxylic groups (pK(1)), varied as a function of nitrogen source, while for E. coli the fourth site concentration (N(4)), corresponding to hydroxyl/amine groups (pK(4)), varied as a function of C:N ratio. Correspondingly, it was found that N(1) was the highest of the four site concentrations for B. brevis and N(4) was the highest for E. coli. The concentrations of the remaining sites showed little variation. Finally, comparison between the titration data and a number of cell surface compositional studies in the literature indicates one distinct difference between the two bacteria is that pK(4) of the Gram-negative E. coli can be attributed to hydroxyl groups while that of the Gram-positive B. brevis can be attributed to amine groups.

OptSSeq: High-throughput sequencing readout of growth enrichment defines optimal gene expression elements for homoethanologenesis

DOE PAGES

Ghosh, Indro Neil; Landick, Robert

2016-07-16

The optimization of synthetic pathways is a central challenge in metabolic engineering. OptSSeq (Optimization by Selection and Sequencing) is one approach to this challenge. OptSSeq couples selection of optimal enzyme expression levels linked to cell growth rate with high-throughput sequencing to track enrichment of gene expression elements (promoters and ribosomebinding sites) from a combinatorial library. OptSSeq yields information on both optimal and suboptimal enzyme levels, and helps identify constraints that limit maximal product formation. Here we report a proof-of-concept implementation of OptSSeq using homoethanologenesis, a two-step pathway consisting of pyruvate decarboxylase (Pdc) and alcohol dehydrogenase (Adh) that converts pyruvate tomore » ethanol and is naturally optimized in the bacterium Zymomonas mobilis. We used OptSSeq to determine optimal gene expression elements and enzyme levels for Z. mobilis Pdc, AdhA, and AdhB expressed in Escherichia coli. By varying both expression signals and gene order, we identified an optimal solution using only Pdc and AdhB. We resolved current uncertainty about the functions of the Fe 2+-dependent AdhB and Zn 2+- dependent AdhA by showing that AdhB is preferred over AdhA for rapid growth in both E. coli and Z. mobilis. Finally, by comparing predictions of growth-linked metabolic flux to enzyme synthesis costs, we established that optimal E. coli homoethanologenesis was achieved by our best pdc-adhB expression cassette and that the remaining constraints lie in the E. coli metabolic network or inefficient Pdc or AdhB function in E. coli. Furthermore, OptSSeq is a general tool for synthetic biology to tune enzyme levels in any pathway whose optimal function can be linked to cell growth or survival.« less

Functional Human α7 Nicotinic Acetylcholine Receptor (nAChR) Generated from Escherichia coli.

PubMed

Tillman, Tommy S; Alvarez, Frances J D; Reinert, Nathan J; Liu, Chuang; Wang, Dawei; Xu, Yan; Xiao, Kunhong; Zhang, Peijun; Tang, Pei

2016-08-26

Human Cys-loop receptors are important therapeutic targets. High-resolution structures are essential for rational drug design, but only a few are available due to difficulties in obtaining sufficient quantities of protein suitable for structural studies. Although expression of proteins in E. coli offers advantages of high yield, low cost, and fast turnover, this approach has not been thoroughly explored for full-length human Cys-loop receptors because of the conventional wisdom that E. coli lacks the specific chaperones and post-translational modifications potentially required for expression of human Cys-loop receptors. Here we report the successful production of full-length wild type human α7nAChR from E. coli Chemically induced chaperones promote high expression levels of well-folded proteins. The choice of detergents, lipids, and ligands during purification determines the final protein quality. The purified α7nAChR not only forms pentamers as imaged by negative-stain electron microscopy, but also retains pharmacological characteristics of native α7nAChR, including binding to bungarotoxin and positive allosteric modulators specific to α7nAChR. Moreover, the purified α7nAChR injected into Xenopus oocytes can be activated by acetylcholine, choline, and nicotine, inhibited by the channel blockers QX-222 and phencyclidine, and potentiated by the α7nAChR specific modulators PNU-120596 and TQS. The successful generation of functional human α7nAChR from E. coli opens a new avenue for producing mammalian Cys-loop receptors to facilitate structure-based rational drug design. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Mature Programs of Study: Examining Policy Implementation at the Local Level. Final Report

ERIC Educational Resources Information Center

Alfeld, Corinne; Bhattacharya, Sharika

2013-01-01

The 2006 Carl D. Perkins Career and Technical Education Act required that all career technical education (CTE) programs offer secondary to postsecondary programs of study (POS), which integrate rigorous academics, offer dual enrollment options, and lead to an industry-recognized degree or credential. Focused on improving students' transition to…

75 FR 37722 - OMB Approvals Under the Paperwork Reduction Act; Technical Amendment

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-30

... since its last update. B. Why is this Technical Amendment Issued as a Final Rule? The information... provisions since the last update of this table. The paperwork burden associated with these new provisions was... amendment updates the table that lists the Office of Management and Budget (OMB) control numbers issued...

United States Air Force Training Line Simulator. Final Report.

ERIC Educational Resources Information Center

Nauta, Franz; Pierce, Michael B.

This report describes the technical aspects and potential applications of a computer-based model simulating the flow of airmen through basic training and entry-level technical training. The objective of the simulation is to assess the impacts of alternative recruit classification and training policies under a wide variety of assumptions regarding…

Final recovery plan of the southwestern willow flycatcher (Empidonax traillii extimus)

Treesearch

Deborah M. Finch; Stephen I. Rothstein; Jon C. Boren; William L. Graf; Jerry L. Holechek; Barbara E. Kus; Robert M. Marshall; Molly M. Pohl; Susan J. Sferra; Mark K. Sogge; Julie C. Stromberg; Bradley A. Valentine; Mary J. Whitfield; Sartor O. Williams

2002-01-01

The Southwestern Willow Flycatcher Recovery Team is composed of a Technical Subgroup (pg. ii), six Implementation Subgroups (Appendix A), and a Tribal Working Group. The Technical Subgroup consists of 14 academic scientists, researchers, and resource managers with a wide range of expertise in avian biology and ecology, southwestern willow flycatcher ecology, cowbird...

Local/State Bilingual Project. 1981-82 Final Technical Report. Appendixes.

ERIC Educational Resources Information Center

Austin Independent School District, TX. Office of Research and Evaluation.

The 1981-82 Local/State Bilingual Program Technical Report addresses the evaluation questions of the Local/State Bilingual Program Evaluation Design. It is organized into six appendixes. Each appendix reports the information collected by a specific measure. Each appendix consists of (1) an instrument description, (2) purpose of the measure, (3)…

77 FR 26444 - Revisions to Final Response To Petition From New Jersey Regarding SO2

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-04

... modeling or other technical analyses and no new analyses were necessary to make the revisions. III. Public... this modeling approach. Therefore, no new technical analyses or any changes to the modeling are...) modeling analysis submitted with the September 2010 petition identified NAAQS violations at receptors in...

77 FR 45282 - NRC Position on the Relationship Between General Design Criteria and Technical Specification...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-07-31

..., are described in the final safety analysis report (FSAR). The staff safety evaluation documents the acceptability of these analyses, and it is the combination of the FSAR analyses and the staff safety evaluation... analysis, maintain their capability to perform their safety functions. Technical Specification Operability...

A Dissemination Model for New Technical Education Programs. Final Report.

ERIC Educational Resources Information Center

Hull, Daniel M.

The Technical Education Research Center-SW has conceived, tested, and refined a model for disseminating newly developed programs and materials throughout the nation. The model performed successfully in the dissemination of more than 50,000 educational units (modules) of Laser/Electro-Optics Technician (LEOT) materials during a four-year period…

75 FR 28499 - Medical Certification Requirements as Part of the Commercial Driver's License (CDL); Technical...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-21

... Driver's License (CDL); Technical, Organizational, and Conforming Amendments AGENCY: Federal Motor... ``medical examiner's license or certificate number'' to refer to the number on a medical examiner's license... examiner qualifies him or her to drive. This inconsistency has been clarified in today's final rule so that...

A Simulation of DNA Sequencing Utilizing 3M Post-It[R] Notes

ERIC Educational Resources Information Center

Christensen, Doug

2009-01-01

An inexpensive and equipment free approach to teaching the technical aspects of DNA sequencing. The activity described requires an instructor with a familiarity of DNA sequencing technology but provides a straight forward method of teaching the technical aspects of sequencing in the absence of expensive sequencing equipment. The final sequence…

Installation and Assimilation of Educational Innovations in Vocational/Technical Programs in Post-Secondary Institutions. Final Report.

ERIC Educational Resources Information Center

Appel, Victor H.; Roueche, John E.

The factors which contribute to the successful installation and assimilation of educational innovations were examined in relation to vocational/technical programs in eight post-secondary institutions. About 555 instructors, administrators, and non-teaching professional staff were surveyed regarding the level and extent of use of individualized…

Final Technical Report on the Institute for Advanced Study in Student Personnel Work.

ERIC Educational Resources Information Center

Callis, Robert

This document reports the planning, implementation, and evaluation of a 9-month institute held at the University of Missouri-Columbia to prepare participants (approximately 20) for leadership positions in student personnel work at junior colleges and technical institutes. The following aspects of the instructional program are discussed and…

Demonstration and Research Program for Teaching Young String Players. Final Report.

ERIC Educational Resources Information Center

Yarborough, William

This report explains a system for rapidly training beginning students in the technical aspects of playing a stringed instrument. The program also affords them a well-rounded, basic knowledge of music. A "numerical" method of notation and concentrated muscular exercises greatly speeded the technical learning process. The daily coordination of ear…

Troubleshooting Instruction in Vocational-Technical Education Via Dynamic Simulation. Final Report.

ERIC Educational Resources Information Center

Finch, Curtis R.

This study was designed to examine the feasibility of using simulation as a means of teaching vocational-technical students to detect and identify malfunctions in selected electrical and mechanical systems. A dynamic simulator was employed which features interchangeable panels and logic that permits the simulation of electrical or mechanical…

Special Project for Research Training in Vocational Education: Research Training Series. Final Report.

ERIC Educational Resources Information Center

Vivian, Neal E.

To upgrade research and research utilization competence of vocational educators, The Center for Vocational and Technical Education and The American Vocational Association planned four 1-week research training programs on: (1) Planning Vocational/Technical Education Programs Based on Manpower Research, (2) Patterns of Career Development as Applied…

78 FR 4030 - Designation of Low-Income Status; Acceptance of Secondary Capital Accounts by Low-Income...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-01-18

... writing that it wishes to accept the designation. The final rule extends an FCU's response time from 30..., nonsubstantive technical amendments to NCUA's requirements for insurance regulation to reflect current agency... requirements for insurance regulation. These technical corrections are necessary to reflect current agency...

SURVEY OF INFORMATION ON VOCATIONAL AND TECHNICAL EDUCATION IN THE STATE OF ILLINOIS. FINAL REPORT.

ERIC Educational Resources Information Center

Corplan Associates, Chicago, IL. Technology Center.

THE BASIC OBJECTIVE OF THE SURVEY WAS TO GATHER INFORMATION HELPFUL IN PLANNING AND DEVELOPING VOCATIONAL AND TECHNICAL EDUCATION PRIMARILY WITHIN THE PUBLIC SCHOOL SYSTEM. OCCUPATIONAL NEEDS WERE IDENTIFIED FROM FORECASTS OF CHANGES IN CURRENT OCCUPATIONS, AN ANALYSIS OF THE IMPLICATIONS OF SCIENTIFIC AND TECHNOLOGICAL DEVELOPMENTS, AND…

75 FR 47211 - Navigation and Navigable Waters; Technical, Organizational, and Conforming Amendments, Sector...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-05

... Puget Sound, WA AGENCY: Coast Guard, DHS. ACTION: Final rule. SUMMARY: This rule makes non-substantive... technical corrections to reflect the renaming of Sector Seattle to Sector Puget Sound as part of the Coast... organizational structure. Sector Seattle has been disestablished and Sector Puget Sound has been established in...

Statewide Curriculum Articulation Project for Vocational-Technical Education in Minnesota. Final Report.

ERIC Educational Resources Information Center

Burger, Laura J.; And Others

The goal of this project was to develop, validate, and utilize a process for vertically articulating curriculum between secondary and post secondary levels of vocational technical education throughout the state of Minnesota. Procedures involved the identification of two areas of staff responsibility: research and development, and service to local…

Delivery of Instruction via Television. Final Report.

ERIC Educational Resources Information Center

Wisconsin State Board of Vocational, Technical, and Adult Education, Madison.

During 1985, the Wisconsin Board of Vocational, Technical, and Adult Education (VTAE) expanded its delivery of instruction via television. Approximately 3,000 persons at 170 sites throughout the United States and Canada participated in the VTAE's Technical School of the Air. In addition to delivering six to eight courses per semester, the network…

O*NET Final Technical Report. Volume I [and] Volume II [and] Volume III.

ERIC Educational Resources Information Center

Peterson, Norman G.; Mumford, Michael D.; Borman, Walter C.; Jeanneret, P. Richard; Fleishman, Edwin A.; Levin, Kerry Y.

This document contains the three volumes of the technical report for development of the prototype of the Occupational Information Network (O*NET), which is intended to replace the "Dictionary of Occupational Titles.""General Introduction" (Norman G. Peterson) presents an overview of O*NET's purpose, content, and structure.…

78 FR 34920 - Technical Amendments To Reflect the Authorizing Legislation of the Institute of Museum and...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-11

... NATIONAL FOUNDATION ON THE ARTS AND THE HUMANITIES Institute of Museum and Library Services 45 CFR... Museum and Library Services AGENCY: Institute of Museum and Library Services (IMLS), NFAH. ACTION: Technical amendment; final rule. SUMMARY: The Institute of Museum and Library amends its grants regulations...

Final report on technical work accomplished under contract NASw-2953

NASA Technical Reports Server (NTRS)

Fredricks, R. W.

1977-01-01

A report is given on the technical work accomplished in the area of plasma physics. The subjects covered are: (1) oblique whistler instabilities, (2) current-limited electron beam injection, (3) three-dimensional ion sound turbulence, (4) theoretical aspects of sounder antenna operation and (5) whistler modes in bow shock structures.

Development of German-English Machine Translation System. Final Technical Report.

ERIC Educational Resources Information Center

Lehmann, Winfred P.; Stachowitz, Rolf A.

This report describes work on a pilot system for a fully automatic, high-quality translation of German scientific and technical text into English and gives the results of an experiment designed to show the system's capability to produce quality mechanical translation. The areas considered were: (1) grammar formalism, mainly involving the addition…

Phenotyping the quality of complex medium components by simple online-monitored shake flask experiments.

PubMed

Diederichs, Sylvia; Korona, Anna; Staaden, Antje; Kroutil, Wolfgang; Honda, Kohsuke; Ohtake, Hisao; Büchs, Jochen

2014-11-07

Media containing yeast extracts and other complex raw materials are widely used for the cultivation of microorganisms. However, variations in the specific nutrient composition can occur, due to differences in the complex raw material ingredients and in the production of these components. These lot-to-lot variations can affect growth rate, product yield and product quality in laboratory investigations and biopharmaceutical production processes. In the FDA's Process Analytical Technology (PAT) initiative, the control and assessment of the quality of critical raw materials is one key aspect to maintain product quality and consistency. In this study, the Respiration Activity Monitoring System (RAMOS) was used to evaluate the impact of different yeast extracts and commercial complex auto-induction medium lots on metabolic activity and product yield of four recombinant Escherichia coli variants encoding different enzymes. Under non-induced conditions, the oxygen transfer rate (OTR) of E. coli was not affected by a variation of the supplemented yeast extract lot. The comparison of E. coli cultivations under induced conditions exhibited tremendous differences in OTR profiles and volumetric activity for all investigated yeast extract lots of different suppliers as well as lots of the same supplier independent of the E. coli variant. Cultivation in the commercial auto-induction medium lots revealed the same reproducible variations. In cultivations with parallel offline analysis, the highest volumetric activity was found at different cultivation times. Only by online monitoring of the cultures, a distinct cultivation phase (e.g. glycerol depletion) could be detected and chosen for comparable and reproducible offline analysis of the yield of functional product. This work proves that cultivations conducted in complex media may be prone to significant variation in final product quality and quantity if the quality of the raw material for medium preparation is not thoroughly checked. In this study, the RAMOS technique enabled a reliable and reproducible screening and phenotyping of complex raw material lots by online measurement of the respiration activity. Consequently, complex raw material lots can efficiently be assessed if the distinct effects on culture behavior and final product quality and quantity are visualized.

The Independent Technical Analysis Process Final Report 2006-2007.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Duberstein, Corey; Ham, Kenneth; Dauble, Dennis

2007-03-01

The Bonneville Power Administration (BPA) contracted with the Pacific Northwest National Laboratory (PNNL) to provide technical analytical support for system-wide fish passage information (BPA Project No. 2006-010-00). The goal of this project was to produce rigorous technical analysis products using independent analysts and anonymous peer reviewers. This project provided an independent technical source for non-routine fish passage analyses while allowing routine support functions to be performed by other well-qualified entities. The Independent Technical Analysis Process (ITAP) was created to provide non-routine analysis for fish and wildlife agencies and tribes in particular and the public in general on matters related tomore » juvenile and adult salmon and steelhead passage through the mainstem hydrosystem. The process was designed to maintain the independence of analysts and reviewers from parties requesting analyses, to avoid potential bias in technical products. The objectives identified for this project were to administer a rigorous, transparent process to deliver unbiased technical assistance necessary to coordinate recommendations for storage reservoir and river operations that avoid potential conflicts between anadromous and resident fish. Seven work elements, designated by numbered categories in the Pisces project tracking system, were created to define and accomplish project goals as follows: (1) 118 Coordination - Coordinate technical analysis and review process: (a) Retain expertise for analyst/reviewer roles. (b) Draft research directives. (c) Send directive to the analyst. (d) Coordinate two independent reviews of the draft report. (e) Ensure reviewer comments are addressed within the final report. (2) 162 Analyze/Interpret Data - Implement the independent aspects of the project. (3) 122 Provide Technical Review - Implement the review process for the analysts. (4) 132 Produce Annual Report - FY06 annual progress report with Pisces Disseminate (5) 161 Disseminate Raw/Summary Data and Results - Post technical products on the ITAP web site. (6) 185-Produce Pisces Status Report - Provide periodic status reports to BPA. (7) 119 Manage and Administer Projects - project/contract administration.« less

Optimization of multi-epitopic HIV-1 recombinant protein expression in prokaryote system and conjugation to mouse DEC-205 monoclonal antibody: implication for in-vivo targeted delivery of dendritic cells

PubMed Central

Rahimi, Roghayeh; Ebtekar, Massoumeh; Moazzeni, Seyed Mohammad; Mostafaie, Ali; Mahdavi, Mehdi

2015-01-01

Objective(s): Multi-epitopic protein vaccines and direction of vaccine delivery to dendritic cells (DCs) are promising approaches for enhancing immune responses against mutable pathogens. Escherichia coli is current host for expression of recombinant proteins, and it is important to optimize expression condition. The aim of this study was the optimization of multi-epitopic HIV-1 tat/pol/gag/env recombinant protein (HIVtop4) expression by E. coli and conjugation of purified protein to anti DEC-205 monoclonal antibody as candidate vaccine. Materials and Methods: In this study, expression was induced in BL21 (DE3) E. coli cells by optimization of induction condition, post induction incubation time, temperature and culture medium formula. Some culture mediums were used for cell culture, and isopropyl-beta-D-thiogalactopyranoside was used for induction of expression. Protein was purified by Ni-NTA column chromatography and confirmed against anti-His antibody in western-blotting. To exploit DCs properties for immunization purposes, recombinant protein chemically coupled to αDEC-205 monoclonal antibody and confirmed against anti-His antibody in western-blotting. Results: The optimum condition for expression was 1 mM IPTG during 4 hr cultures in 2XYT medium, and final protein produced in soluble form. Conjugation of purified protein to αDEC-205 antibody resulted in smears of protein: antibodies conjugate in different molecular weights. Conclusion: The best cultivation condition for production of HIVtop4 protein is induction by 1 mM IPTG during 4 hr in 2XYT medium. The final concentration of purified protein was 500 µg/ml. PMID:25810888

Back to basics: pBR322 and protein expression systems in E. coli.

PubMed

Balbás, Paulina; Bolívar, Francisco

2004-01-01

The extensive variety of plasmid-based expression systems in E. coli resulted from the fact that there is no single strategy for achieving maximal expression of every cloned gene. Although a number of strategies have been implemented to deal with problems associated to gene transcription and translation, protein folding, secretion, location, posttranslational modifications, particularities of different strains, and the like and more integrated processes have been developed, the basic plasmid-borne elements and their interaction with the particular host strain will influence the overall expression system and final productivity. Plasmid vector pBR322 is a well-established multipurpose cloning vector in laboratories worldwide, and a large number of derivatives have been created for specific applications and research purposes, including gene expression in its natural host, E. coli, and few other bacteria. The early characterization of the molecule, including its nucleotide sequence, replication and maintenance mechanisms, and determination of its coding regions, accounted for its success, not only as a universal cloning vector, but also as a provider of genes and an origin of replication for other intraspecies vectors. Since the publication of the aforementioned reviews, novel discoveries pertaining to these issues have appeared in the literature that deepen the understanding of the plasmid's features, behavior, and impact in gene expression systems, as well as some important strain characteristics that affect plasmid replication and stability. The objectives of this review include updating and discussing the new information about (1) the replication and maintenance of pBR322; (2) the host-related modulation mechanisms of plasmid replication; (3) the effects of growth rate on replication control, stability, and recombinant gene expression; (4) ways for plasmid amplification and elimination. Finally, (5) a summary of novel ancillary studies about pBR322 is presented.

[Expression optimization and characterization of Tenebrio molitor antimicrobiol peptides TmAMP1m in Escherichia coli].

PubMed

Alimu, Reyihanguli; Mao, Xinfang; Liu, Zhongyuan

2013-06-01

To improve the expression level of tmAMP1m gene from Tenebrio molitor in Escherichia coli, we studied the effects of expression level and activity of the fusion protein HIS-TmAMP1m by conditions, such as culture temperature, inducing time and the final concentration of inductor Isopropyl beta-D-thiogalactopyranoside (IPTG). We analyzed the optimum expression conditions by Tricine-SDS-PAGE electrophoresis, meanwhile, detected its antibacterial activity by using agarose cavity diffusion method. The results suggest that when inducing the recombinant plasmid with a final IPTG concentration of 0.1 mmol/L at 37 degrees C for 4 h, there was the highest expression level of fusion protein HIS-TmAMP1m in Escherichia coli. Under these conditions, the expression of fusion protein accounted for 40% of the total cell lysate with the best antibacterial activity. We purified the fusion protein HIS-TmAMPlm with nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography matrices. Western blotting analysis indicates that the His monoclonal antibody could be specifically bound to fusion protein HIS-TmAMPlm. After expression by inducing, the fusion protein could inhibit the growth of host cell transformed by pET30a-tmAMP1m. The fusion protein HIS-TmAMP1m had better stability and remained higher antibacterial activities when incubated at 100 degrees C for 10 h, repeated freeze thawing at -20 degrees C, dissolved in strong acid and alkali, or treated by organic solvents and protease. Moreover, the minimum inhibitory concentration results demonstrated that the fusion protein HIS-TmAMP1m has a good antibacterial activity against Staphylococcus aureus, Staphylococcus sp., Corynebacterium glutamicum, Bacillus thuringiensis, Corynebacterium sp. This study laid the foundation to promote the application of insect antimicrobial peptides and further research.

Site Operator technical report. Final report (1992--1996)

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

1996-12-01

The Southern California Edison Company (SCE) and the US Department of Energy (DOE) entered into cooperative agreement No. DE-FC07-91ID13077 on August 23, 1991, which expired on August 3, 1996. This cooperative agreement provided SCE with DOE cofunding for participation in the DOE`s Electric and Hybrid Vehicle Site Operator Program. In return, SCE provided the DOE with quarterly progress reports which include operating and maintenance data for the electric (EVs) vehicles in SCE`s fleet. Herein is SCE`s final report for the 1992 to 1996 agreement period. As of September 1, 1996 the SCE fleet had 65 electric vehicles in service. Amore » total of 578,200 miles had been logged. During the agreement period, SCE sent the DOE a total of 19 technical reports (Appendix B). This report summarizes the technical achievements which took place during a long, productive and rewarding, relationship with the DOE.« less

7 CFR 1944.419 - Final grantee evaluation.

Code of Federal Regulations, 2010 CFR

2010-01-01

...) PROGRAM REGULATIONS (CONTINUED) HOUSING Self-Help Technical Assistance Grants § 1944.419 Final grantee... Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS..., application, this regulation, and any amendments. (a) This is a quantitative evaluation of the grantee to...

ECLSS medical support activities

NASA Technical Reports Server (NTRS)

Crump, William J.; Kilgore, Melvin V., Jr.

1991-01-01

During the period from April 10, 1990 to April 9, 1991, the Consortium for the Space Life Sciences provided technical assistance to the NASA/MSFC water recovery efforts. This assistance was in the form of literature reviews, technical recommendations, and presentations. This final report summarizes the activities completed during this period and identifies those areas requiring additional efforts. The tasks which the University of Alabama in Huntsville (UAH) water recovery team addressed were either identified by MSFC technical representatives or chosen from those outlined in the subject statement of work.

Switching gears and changing lanes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hammock, Earlene C.

2002-01-01

From musing over Shakespeare's fine lines and metaphors to teaching technical writing and editing, and finally, to cranking out scientific and technical documents, writing articles, and editing publications-how'd a nice girl like me end up in a place like this? Twice, after having prepared for an academic career of teaching and research, I found myself in a technical communications position-first, at the University of Texas at El Paso College of Engineering and later, at Los Alamos National Laboratory. What happened?

NASA/DOD Aerospace Knowledge Diffusion Research Project. Paper 32: A new era in international technical communication: American-Russian collaboration

NASA Technical Reports Server (NTRS)

Flammia, Madelyn; Barclay, Rebecca O.; Pinelli, Thomas E.; Keene, Michael L.; Burger, Robert H.; Kennedy, John M.

1993-01-01

Until the recent dissolution of the Soviet Union, the Communist Party exerted a strict control of access to and dissemination of scientific and technical information (STI). This article presents models of the Soviet-style information society and the Western-style information society and discusses the effects of centralized governmental control of information on Russian technical communication practices. The effects of political control on technical communication are then used to interpret the results of a survey of Russian and U.S. aerospace engineers and scientists concerning the time devoted to technical communication, their collaborative writing practices and their attitudes toward collaboration, the kinds of technical documents they produce and use, their views regarding the appropriate content for an undergraduate technical communication course, and their use of computer technology. Finally, the implications of these findings for future collaboration between Russian and U.S. engineers and scientists are examined.

NASA/DoD Aerospace Knowledge Diffusion Research Project. XXXII - A new era in international technical communication: American-Russian collaboration

NASA Technical Reports Server (NTRS)

Flammia, Madelyn; Barclay, Rebecca O.; Pinelli, Thomas E.; Keene, Michael L.; Burger, Robert H.; Kennedy, John M.

1993-01-01

Until the recent dissolution of the Soviet Union, the Communist Party exerted a strict control of access to and dissemination of scientific and technical information. This article presents models of the Soviet-style information society and the Western-style information society and discusses the effects of centralized governmental control of information on Russian technical communication practices. The effects of political control on technical communication are then used to interpret the results of a survey of Russian and U.S. aerospace engineers and scientists concerning the time devoted to technical communication, their collaborative writing practices and their attitudes toward collaboration, the kinds of technical documents they produce and use, their views regarding the appropriate content for an undergraduate technical communication course, and their use of computer technology. Finally, the implications of these findings for future collaboration between Russian and U.S. engineers and scientists are examined.

Identification of ethanol tolerant outer membrane proteome reveals OmpC-dependent mechanism in a manner of EnvZ/OmpR regulation in Escherichia coli.

PubMed

Zhang, Dan-Feng; Ye, Jin-Zhou; Dai, Hong-Hou; Lin, Xiang-Min; Li, Hui; Peng, Xuan-Xian

2018-05-15

Ethanol is an efficient disinfectant, but long-term and wide usage of ethanol leads to microbial tolerance. Bacteria with the tolerance are widely identified. However, mechanisms of the tolerance are not elucidated. To explore the mechanisms of outer membrane (OM) proteins underlying ethanol tolerance in bacteria, functional proteomic methodologies were utilized to characterize OM proteins of E. coli suddenly exposed to 3.125% ethanol. Of eleven proteins altered significantly, seven were OM proteins, in which LamB, FadL and OmpC were up-regulated, and OmpT, OmpF, Tsx and OmpA were down-regulated. The alterations were validated using Western blot. Then, functional characterization of the altered abundance of OM proteins was investigated in gene-deleted and gene-complemented mutants cultured in 1.56-6.25% ethanol. Higher inhibiting rate was detected in ΔompC than ΔlamB and ΔompA, but no difference was found between Δtsx, ΔompF, ΔfadL or ΔompT and control. Furthermore, EnvZ/OmpR two-component signal transduction system, which regulates OmpC and OmpF expression, was determined to participate in the tolerance. Finally, our results show that absence of envZ, ompR or ompC and ompA led to elevated and reduced intracellular ethanol, respectively. These findings indicate EnvZ-dependent phosphotransfer signaling pathway of the OmpR-mediated expression of OmpC plays a crucial role in ethanol tolerance. Ethanol tolerance is an adaptation strategy of bacteria. In the present study, we used the proteomic approaches involving 2-DE and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) to determined outer membrane (OM) protein changes in E. coli K-12 after 2 h of 1/2 MIC of ethanol exposure. Under ethanol stress, seven differential OM proteins were found, which were validated by Western blot. Functions of these seven OM proteins were compared using their genetically modified strains. Furthermore, the role of EnvZ/OmpR two-component signal transduction system was identified in ethanol tolerance of E. coli. Finally, Loss of ompC, envZ or ompR increases intracellular ethanol, while absence of ompA reduces reversal effect. This is the first report of OM proteomics in E. coli exposed to ethanol. Our findings reveal an unknown OmpC-dependent mechanism of ethanol tolerance in a manner of EnvZ/OmpR regulation. Copyright © 2018 Elsevier B.V. All rights reserved.

Preparation and Characterization of Antimicrobial Films Based on LDPE/Ag Nanoparticles with Potential Uses in Food and Health Industries.

PubMed

Olmos, Dania; Pontes-Quero, Gloria María; Corral, Angélica; González-Gaitano, Gustavo; González-Benito, Javier

2018-01-24

In this work, the antimicrobial effect of silver nanoparticles in polyethylene based nanocomposites has been investigated using a non-conventional processing method to produce homogeneous materials. High energy ball milling under cryogenic conditions was used to achieve a powder of well-blended low-density polyethylene and commercial silver nanoparticles. The final composites in the form of films were obtained by hot pressing. The effect of various silver nanoparticles content (0, 0.5, 1 and 2 wt %) on the properties of low-density polyethylene and the antimicrobial effectiveness of the composite against DH5α Escherichia coli were studied. The presence of silver nanoparticles did not seem to affect the surface energy and thermal properties of the materials. Apart from the inhibition of bacterial growth, slight changes in the aspect ratio of the bacteria with the content of particles were observed, suggesting a direct relationship between the presence of silver nanoparticles and the proliferation of DH5α E. coli ( Escherichia coli ) cells. Results indicate that these materials may be used to commercially produce antimicrobial polymers with potential applications in the food and health industries.

Preparation and Characterization of Antimicrobial Films Based on LDPE/Ag Nanoparticles with Potential Uses in Food and Health Industries

PubMed Central

Pontes-Quero, Gloria María; Corral, Angélica

2018-01-01

In this work, the antimicrobial effect of silver nanoparticles in polyethylene based nanocomposites has been investigated using a non-conventional processing method to produce homogeneous materials. High energy ball milling under cryogenic conditions was used to achieve a powder of well-blended low-density polyethylene and commercial silver nanoparticles. The final composites in the form of films were obtained by hot pressing. The effect of various silver nanoparticles content (0, 0.5, 1 and 2 wt %) on the properties of low-density polyethylene and the antimicrobial effectiveness of the composite against DH5α Escherichia coli were studied. The presence of silver nanoparticles did not seem to affect the surface energy and thermal properties of the materials. Apart from the inhibition of bacterial growth, slight changes in the aspect ratio of the bacteria with the content of particles were observed, suggesting a direct relationship between the presence of silver nanoparticles and the proliferation of DH5α E. coli (Escherichia coli) cells. Results indicate that these materials may be used to commercially produce antimicrobial polymers with potential applications in the food and health industries. PMID:29364193

The NAG Sensor NagC Regulates LEE Gene Expression and Contributes to Gut Colonization by Escherichia coli O157:H7.

PubMed

Le Bihan, Guillaume; Sicard, Jean-Félix; Garneau, Philippe; Bernalier-Donadille, Annick; Gobert, Alain P; Garrivier, Annie; Martin, Christine; Hay, Anthony G; Beaudry, Francis; Harel, Josée; Jubelin, Grégory

2017-01-01

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 are human pathogens responsible for bloody diarrhea and renal failures. EHEC employ a type 3 secretion system to attach directly to the human colonic epithelium. This structure is encoded by the locus of enterocyte effacement (LEE) whose expression is regulated in response to specific nutrients. In this study, we show that the mucin-derived sugars N-acetylglucosamine (NAG) and N-acetylneuraminic acid (NANA) inhibit EHEC adhesion to epithelial cells through down-regulation of LEE expression. The effect of NAG and NANA is dependent on NagC, a transcriptional repressor of the NAG catabolism in E. coli . We show that NagC is an activator of the LEE1 operon and a critical regulator for the colonization of mice intestine by EHEC. Finally, we demonstrate that NAG and NANA as well as the metabolic activity of Bacteroides thetaiotaomicron affect the in vivo fitness of EHEC in a NagC-dependent manner. This study highlights the role of NagC in coordinating metabolism and LEE expression in EHEC and in promoting EHEC colonization in vivo .

The NAG Sensor NagC Regulates LEE Gene Expression and Contributes to Gut Colonization by Escherichia coli O157:H7

PubMed Central

Le Bihan, Guillaume; Sicard, Jean-Félix; Garneau, Philippe; Bernalier-Donadille, Annick; Gobert, Alain P.; Garrivier, Annie; Martin, Christine; Hay, Anthony G.; Beaudry, Francis; Harel, Josée; Jubelin, Grégory

2017-01-01

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 are human pathogens responsible for bloody diarrhea and renal failures. EHEC employ a type 3 secretion system to attach directly to the human colonic epithelium. This structure is encoded by the locus of enterocyte effacement (LEE) whose expression is regulated in response to specific nutrients. In this study, we show that the mucin-derived sugars N-acetylglucosamine (NAG) and N-acetylneuraminic acid (NANA) inhibit EHEC adhesion to epithelial cells through down-regulation of LEE expression. The effect of NAG and NANA is dependent on NagC, a transcriptional repressor of the NAG catabolism in E. coli. We show that NagC is an activator of the LEE1 operon and a critical regulator for the colonization of mice intestine by EHEC. Finally, we demonstrate that NAG and NANA as well as the metabolic activity of Bacteroides thetaiotaomicron affect the in vivo fitness of EHEC in a NagC-dependent manner. This study highlights the role of NagC in coordinating metabolism and LEE expression in EHEC and in promoting EHEC colonization in vivo. PMID:28484684

A Sulfurtransferase Is Essential for Activity of Formate Dehydrogenases in Escherichia coli*

PubMed Central

Thomé, Rémi; Gust, Alexander; Toci, René; Mendel, Ralf; Bittner, Florian; Magalon, Axel; Walburger, Anne

2012-01-01

l-Cysteine desulfurases provide sulfur to several metabolic pathways in the form of persulfides on specific cysteine residues of an acceptor protein for the eventual incorporation of sulfur into an end product. IscS is one of the three Escherichia coli l-cysteine desulfurases. It interacts with FdhD, a protein essential for the activity of formate dehydrogenases (FDHs), which are iron/molybdenum/selenium-containing enzymes. Here, we address the role played by this interaction in the activity of FDH-H (FdhF) in E. coli. The interaction of IscS with FdhD results in a sulfur transfer between IscS and FdhD in the form of persulfides. Substitution of the strictly conserved residue Cys-121 of FdhD impairs both sulfur transfer from IscS to FdhD and FdhF activity. Furthermore, inactive FdhF produced in the absence of FdhD contains both metal centers, albeit the molybdenum cofactor is at a reduced level. Finally, FdhF activity is sulfur-dependent, as it shows reversible sensitivity to cyanide treatment. Conclusively, FdhD is a sulfurtransferase between IscS and FdhF and is thereby essential to yield FDH activity. PMID:22194618

Fed-batch culture of Escherichia coli for L-valine production based on in silico flux response analysis.

PubMed

Park, Jin Hwan; Kim, Tae Yong; Lee, Kwang Ho; Lee, Sang Yup

2011-04-01

We have previously reported the development of a 100% genetically defined engineered Escherichia coli strain capable of producing L-valine from glucose with a high yield of 0.38 g L-valine per gram glucose (0.58 mol L-valine per mol glucose) by batch culture. Here we report a systems biological strategy of employing flux response analysis in bioprocess development using L-valine production by fed-batch culture as an example. Through the systems-level analysis, the source of ATP was found to be important for efficient L-valine production. There existed a trade-off between L-valine production and biomass formation, which was optimized for the most efficient L-valine production. Furthermore, acetic acid feeding strategy was optimized based on flux response analysis. The final fed-batch cultivation strategy allowed production of 32.3 g/L L-valine, the highest concentration reported for E. coli. This approach of employing systems-level analysis of metabolic fluxes in developing fed-batch cultivation strategy would also be applicable in developing strategies for the efficient production of other bioproducts. Copyright © 2010 Wiley Periodicals, Inc.

L-Tryptophan Production in Escherichia coli Improved by Weakening the Pta-AckA Pathway

PubMed Central

Liu, Lina; Duan, Xuguo; Wu, Jing

2016-01-01

Acetate accumulation during the fermentation process of Escherichia coli FB-04, an L-tryptophan production strain, is detrimental to L-tryptophan production. In an initial attempt to reduce acetate formation, the phosphate acetyltransferase gene (pta) from E. coli FB-04 was deleted, forming strain FB-04(Δpta). Unfortunately, FB-04(Δpta) exhibited a growth defect. Therefore, pta was replaced with a pta variant (pta1) from E. coli CCTCC M 2016009, forming strain FB-04(pta1). Pta1 exhibits lower catalytic capacity and substrate affinity than Pta because of a single amino acid substitution (Pro69Leu). FB-04(pta1) lacked the growth defect of FB-04(Δpta) and showed improved fermentation performance. Strain FB-04(pta1) showed a 91% increase in L-tryptophan yield in flask fermentation experiments, while acetate production decreased by 35%, compared with its parent FB-04. Throughout the fed-batch fermentation process, acetate accumulation by FB-04(pta1) was slower than that by FB-04. The final L-tryptophan titer of FB-04(pta1) reached 44.0 g/L, representing a 15% increase over that of FB-04. Metabolomics analysis showed that the pta1 genomic substitution slightly decreased carbon flux through glycolysis and significantly increased carbon fluxes through the pentose phosphate and common aromatic pathways. These results indicate that this strategy enhances L-tryptophan production and decreases acetate accumulation during the L-tryptophan fermentation process. PMID:27348810

Microbiological testing of raw, boxed beef in the context of hazard analysis critical control point at a high-line-speed abattoir.

PubMed

Jericho, K W; Kozub, G C; Gannon, V P; Taylor, C M

2000-12-01

The efficacy of cold storage of raw, bagged, boxed beef was assessed microbiologically at a high-line-speed abattoir (270 carcasses per h). At the time of this study, plant management was in the process of creating a hazard analysis critical control point plan for all processes. Aerobic bacteria, coliforms, and type 1 Escherichia coli were enumerated (5 by 5-cm excision samples, hydrophobic grid membrane filter technology) before and after cold storage of this final product produced at six fabrication tables. In addition, the temperature-function integration technique (TFIT) was used to calculate the potential number of generations of E. coli during the first 24 or 48 h of storage of the boxed beef. Based on the temperature histories (total of 60 boxes, resulting from 12 product cuts, five boxes from each of two fabrication tables on each of 6 sampling days, and six types of fabrication tables), TFIT did not predict any growth of E. coli (with or without lag) for the test period. This was verified by E. coli mean log10 values of 0.65 to 0.42 cm2 (P > 0.05) determined by culture before and after the cooling process, respectively. Counts of aerobic bacteria and coliforms were significantly reduced (P < 0.001 and P < 0.05, respectively) during the initial period of the cooling process. There were significant microbiological differences (P < 0.05) between table-cut units.

Water quality at points-of-use in the Galapagos Islands.

PubMed

Gerhard, William A; Choi, Wan Suk; Houck, Kelly M; Stewart, Jill R

2017-04-01

Piped drinking water is often considered a gold standard for protecting public health but research is needed to explicitly evaluate the effect of centralized treatment systems on water quality in developing world settings. This study examined the effect of a new drinking water treatment plant (DWTP) on microbial drinking water quality at the point-of-use on San Cristobal Island, Galapagos using fecal indicator bacteria total coliforms and Escherichia coli. Samples were collected during six collection periods before and after operation of the DWTP began from the freshwater sources (n=4), the finished water (n=6), and 50 sites throughout the distribution system (n=287). This study found that there was a significant decrease in contamination by total coliforms (two orders of magnitude) and E. coli (one order of magnitude) after DWTP operation began (p<0.001). However, during at least one post-construction collection cycle, total coliforms and E. coli were still found at 66% and 28% of points-of-use (n=50), respectively. During the final collection period, conventional methods were augmented with human-specific Bacteroides assays - validated herein - with the goal of elucidating possible microbial contamination sources. Results show that E. coli contamination was not predictive of contamination by human wastes and suggests that observed indicator bacteria contamination may have environmental origins. Together these findings highlight the necessity of a holistic approach to drinking water infrastructure improvements in order to deliver high quality water through to the point-of-use. Copyright © 2017 Elsevier GmbH. All rights reserved.

Identification of bottlenecks in Escherichia coli engineered for the production of CoQ(10).

PubMed

Cluis, Corinne P; Ekins, Andrew; Narcross, Lauren; Jiang, Heng; Gold, Nicholas D; Burja, Adam M; Martin, Vincent J J

2011-11-01

In this work, Escherichia coli was engineered to produce a medically valuable cofactor, coenzyme Q(10) (CoQ(10)), by removing the endogenous octaprenyl diphosphate synthase gene and functionally replacing it with a decaprenyl diphosphate synthase gene from Sphingomonas baekryungensis. In addition, by over-expressing genes coding for rate-limiting enzymes of the aromatic pathway, biosynthesis of the CoQ(10) precursor para-hydroxybenzoate (PHB) was increased. The production of isoprenoid precursors of CoQ(10) was also improved by the heterologous expression of a synthetic mevalonate operon, which permits the conversion of exogenously supplied mevalonate to farnesyl diphosphate. The over-expression of these precursors in the CoQ(10)-producing E. coli strain resulted in an increase in CoQ(10) content, as well as in the accumulation of an intermediate of the ubiquinone pathway, decaprenylphenol (10P-Ph). In addition, the over-expression of a PHB decaprenyl transferase (UbiA) encoded by a gene from Erythrobacter sp. NAP1 was introduced to direct the flux of DPP and PHB towards the ubiquinone pathway. This further increased CoQ(10) content in engineered E. coli, but decreased the accumulation of 10P-Ph. Finally, we report that the combined over-production of isoprenoid precursors and over-expression of UbiA results in the decaprenylation of para-aminobenzoate, a biosynthetic precursor of folate, which is structurally similar to PHB. Copyright © 2011 Elsevier Inc. All rights reserved.

Comparison of methods for recovery of Escherichia coli and Staphylococcus aureus from seeded laundry fabrics.

PubMed Central

Cody, H J; Smith, P F; Blaser, M J; LaForce, F M; Wang, W L

1984-01-01

To assess the effect of laundry procedures on fabric-associated bacteria, a standard method of enumeration is needed. We evaluated six methods for enumeration of Escherichia coli and Staphylococcus aureus seeded (10(2) and 10(5) CFU/100 cm2 of fabric area) onto sterilized hospital sheets and terry . Two methods involved maceration of seeded swatches in broth followed by passage of the broth through a 0.45-micron-pore-size, 47-mm-diameter filter membrane. Three methods involved agitation of seeded swatches in broth with a paint shaker and membrane filtration of the broth to recover eluted bacterial cells, and the final method involved direct enumeration of cells on fabrics by overlaying seeded swatches with agar containing triphenyltetrazolium chloride as an indicator. The most convenient recovery method employed a 90-s agitation followed by serial dilution of broths and membrane filtration. This method provided 44/57% (low seed/high seed) recovery of E. coli from sheets and 133/31% from terry and 34/74% recovery of S. aureus from sheets and 58/57% from terry . Although maceration provided similar recovery of E. coli and S. aureus, it is a less-practical method. The direct enumeration method was ineffective for enumerating gram-positive bacteria. We conclude that either the agitation or maceration method used enumerated the seeded bacteria to within 1 log10 of their expected number and can be used to assess the bactericidal effectiveness of various steps in the laundering process. PMID:6378092

Construction of an alternative glycerol-utilization pathway for improved β-carotene production in Escherichia coli.

PubMed

Guo, Jin-Ying; Hu, Kun-Le; Bi, Chang-Hao; Li, Qing-Yan; Zhang, Xue-Li

2018-05-11

Glycerol, which is an inevitable by-product of biodiesel production, is an ideal carbon source for the production of carotenoids due to its low price, good availability and chemically reduced status, which results in a low requirement for additional reducing equivalents. In this study, an alternative carbon-utilization pathway was constructed in Escherichia coli to enable more efficient β-carotene production from glycerol. An aldehyde reductase gene (alrd) and an aldehyde dehydrogenase gene (aldH) from Ralstonia eutropha H16 were integrated into the E. coli chromosome to form a novel glycerol-utilization pathway. The β-carotene specific production value was increased by 50% after the introduction of alrd and aldH. It was found that the glycerol kinase gene (garK), alrd and aldH were the bottleneck of the alternative glycerol metabolic pathway, and modulation of garK gene with an mRS library further increased the β-carotene specific production value by 13%. Finally, co-modulation of genes in the introduced aldH-alrd operon led to 86% more of β-carotene specific production value than that of the strain without the alternative glycerol-utilization pathway and the glycerol-utilization rate was also increased. In this work, β-carotene production of E. coli was significantly improved by constructing and optimizing an alternative glycerol-utilization pathway. This strategy can potentially be used to improve the production of other isoprenoids using glycerol as a cheap and abundant substrate, and therefore has industrial relevance.

Communication Platform Payload Definition (CPPD) study. Volume 2: Technical report

NASA Technical Reports Server (NTRS)

Hunter, E. M.; Driggers, T.; Jorasch, R.

1986-01-01

This is Volume 2 (Technical Report) of the Ford Aerospace & Communications Corporation Final Report for the Communication Platform Payload Definition (CPPD) Study program conducted for NASA Lewis Research Center under contract No. NAS3-24235. This report presents the results of the study effort leading to five potential platform payloads to service CONUS and WARC Region 2 traffic demand as projected to the year 2008. The report addresses establishing the data bases, developing service aggregation scenarios, selecting and developing 5 payload concepts, performing detailed definition of the 5 payloads, costing them, identifying critical technology, and finally comparing the payloads with each other and also with non-aggregated equivalent services.

Best Practices for Continuous Monitoring of Temperature and Flow in Wadeable Streams (Final Report)

EPA Science Inventory

This final report is a technical "best practices" document describing sensor deployment for and collection of continuous temperature and flow data at ungaged sites in wadeable streams. This document addresses questions related to equipment needs; configuration, placement, and ins...

Hazardous Waste Management System - Identification and Listing of Hazardous Waste - Recycled Used Oil Management Standards - Federal Register Notice, May 3, 1993

EPA Pesticide Factsheets

This action corrects several technical errors and provides clarifying amendments to the final recycled used oil management standards rule. The final rule was published on September 10, 1992 (57 FR 41566).

75 FR 6839 - Technical Service Provider Assistance

Federal Register 2010, 2011, 2012, 2013, 2014

2010-02-12

... Executive Order 12866, the Office of Management and Budget determined that this final rule is not a... or are eligible to participate in conservation programs to help them make land management decisions...: Covered Programs. The interim final rule incorporated reference to the Agricultural Management Assistance...

Charlotte Circle Outreach. Final Report.

ERIC Educational Resources Information Center

Calhoun, Mary Lynne; Rose, Terry L.; Prendergast, Donna

This final report details the activities of the Charlotte Circle Outreach, a program designed to provide technical assistance and training to early intervention programs offering services to infants and young children with substantial disabilities, ages birth through two years. This mission was accomplished through cooperative planning with…

Determination of the Biologically Relevant Sampling Depth for Terrestrial and Aquatic Ecological Risk Assessments (Final Report)

EPA Science Inventory

The Ecological Risk Assessment Support Center (ERASC) announced the release of the final report, Determination of the Biologically Relevant Sampling Depth for Terrestrial and Aquatic Ecological Risk Assessments. This technical paper provides defensible approximations fo...

Properties of uvrE mutants of Escherichia coli K12. Part 1. Effects of uv irradiation on DNA metabolism

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vansluis, C.A.; Mattern, I.E.; Paterson, M.C.

1974-01-01

Escherichia coli K12 uvrE is a mutator strain which is highly sensitive to ultraviolet radiation. In an attempt to determine the underlying molecular basis for the UV sensitivity, a mutant and an isogenic wild type strain were compared with regard to several metabolic responses to 254 nm radiation. The introduction of single strand breaks into intracellular DNA after irradiation is normal; however, the rate of excision of pyrimidine dimers as well as of DNA degradation and final rejoining of the strand breaks is lower in the mutant as compared to the repair proficient strain. These data suggest that the uvrEmore » gene product may be involved in a reaction between the incision and excision steps in the excision repair process. (Author) (GRA)« less

Structure of Escherichia coli AdhP (ethanol-inducible dehydrogenase) with bound NAD.

PubMed

Thomas, Leonard M; Harper, Angelica R; Miner, Whitney A; Ajufo, Helen O; Branscum, Katie M; Kao, Lydia; Sims, Paul A

2013-07-01

The crystal structure of AdhP, a recombinantly expressed alcohol dehydrogenase from Escherichia coli K-12 (substrain MG1655), was determined to 2.01 Å resolution. The structure, which was solved using molecular replacement, also included the structural and catalytic zinc ions and the cofactor nicotinamide adenine dinucleotide (NAD). The crystals belonged to space group P21, with unit-cell parameters a = 68.18, b = 118.92, c = 97.87 Å, β = 106.41°. The final R factor and Rfree were 0.138 and 0.184, respectively. The structure of the active site of AdhP suggested a number of residues that may participate in a proton relay, and the overall structure of AdhP, including the coordination to structural and active-site zinc ions, is similar to those of other tetrameric alcohol dehydrogenase enzymes.

A Program of Technical Assistance to Industry in Twenty-Six Mississippi Counties and the Choctaw Indian Reservation. Final Report.

ERIC Educational Resources Information Center

Phillips, Jane; Sewell, Charles

A broad technical assistance program has been established in 25 EDA (Economic Development Administration) contract counties and on the Choctaw Indian Reservation Nashoba County) to stimulate new job opportunities by solving operational problems which limit the expansion and diversification of existing industry; professional services in evaluating…

For Work-Force Training, a Plan to Give College Credit Where It's Due

ERIC Educational Resources Information Center

Sander, Libby

2008-01-01

After nearly three years of planning, Ohio's higher-education officials are finalizing an ambitious program to grant college credit for some technical courses offered at the state's adult-education centers. The program, called the Career-Technical Credit Transfer, is the latest in a string of state efforts to more closely link work-force training…

Improving the Dissemination of Scientific and Technical Information: A Practitioner's Guide to Innovation. (Final Report).

ERIC Educational Resources Information Center

Capital Systems Group, Inc., Rockville, MD.

The aim of this guide is to alert persons with an operational interest in scientific communication to new ideas, techniques, and equipment in the field of communication media and publications. The focus is on the dissemination of scientific information via the technical journal or its equivalent. Secondary dissemination of information such as…

Unified Technical Concepts--Phase II. Expand Application to Industrial Technologies and Adult Education. Final Report.

ERIC Educational Resources Information Center

Technical Education Research Center, Waco, TX.

A project was conducted to develop a laboratory-based instructional system in physics for two-year technician programs that emphasizes both the analogies between basic physical principles and the applications of the principles in modern technology. The Unified Technical Concepts (UTC) system that was developed is (1) a reorganization of physics…

THE DEVELOPMENT OF ACHIEVEMENT MEASURES FOR TRADE AND TECHNICAL EDUCATION. PROGRESS REPORT NUMBER THREE.

ERIC Educational Resources Information Center

BALDWIN, THOMAS S.

DURING THIS PERIOD FROM SEPTEMBER 1 THROUGH NOVEMBER 30, 1966, 35 FIELD CONSULTANTS COMPLETED ANALYSIS OF THEIR INDIVIDUAL TRADE AND TECHNICAL CURRICULUMS. THESE ANALYSES WERE DEVELOPED INTO AN OUTLINE TO SERVE AS A GUIDE FOR DEVELOPING ACHIEVEMENT TESTS. THE FINAL OUTLINE WAS DIVIDED INTO AS MANY DIFFERENT AREAS AS THE CONSULTANTS FELT NECESSARY…

Word Lists to Simplify Vocabulary of Technical Information. Final Report.

ERIC Educational Resources Information Center

Kincaid, J. Peter; And Others

This report describes eight word lists developed for use as part of the computer readability editing system (CRES), which was developed to serve as an author's aid in improving the ease of comprehending Navy technical manuals and training materials. The system has features which flag uncommon and misspelled words and long sentences, suggest simple…

77 FR 1889 - Drivers of CMVs: Restricting the Use of Cellular Phones; Technical Amendment

Federal Register 2010, 2011, 2012, 2013, 2014

2012-01-12

... DEPARTMENT OF TRANSPORTATION Federal Motor Carrier Safety Administration 49 CFR Part 391 [Docket No. FMCSA-2010-0096] RIN 2126-AB29 Drivers of CMVs: Restricting the Use of Cellular Phones; Technical... Cellular Phones final rule (76 FR 75470) had a clerical error in Sec. 391.15(f)(1) that stated ``paragraph...

27 Years of Impact: Vocational-Technical Education in Ohio. Final Annual Report under Federal Law.

ERIC Educational Resources Information Center

Ohio State Council on Vocational Education, Westerville.

The Ohio Council on Vocational Education (OCOVE) was created to strengthen the career, vocational, and technical education services provided for Ohioans as a practical, efficient, and sure way to enhance the competitiveness of individual workers and the state and national economy. Some of the accomplishments of OCOVE during its 27 years of…

Vocational-Technical Education Interface with Ohio's High Technology Business and Industrial Sector. Final Report.

ERIC Educational Resources Information Center

McCormick, Robert W.

This study explored the relationship of vocational-technical educational institutions in Ohio with business and industry using high-technology applications. The study attempted to determine what high-technology applications will be adopted by Ohio's business and industry in the next 5 years, what experience the schools have had in working with…

National Institute of Statistical Sciences Data Confidentiality Technical Panel: Final Report. NCES 2011-608

ERIC Educational Resources Information Center

Karr, Alan

2011-01-01

NCES asked the National Institute of Statistical Sciences (NISS) to convene a technical panel of survey and policy experts to examine the NCES current and planned data dissemination strategies for confidential data with respect to: mandates and directives that NCES make data available; current and prospective technologies for protecting and…

[Municipalities as a Model for New Careers and Redirection of Vocational-Technical Education Programs.] Final Report.

ERIC Educational Resources Information Center

Institute for Local Self Government, Berkeley, CA.

To meet the manpower needs of local governments, the model developed for this project redirects national and technical education toward new careers programs. Designed by task forces of professional personnel, the model utilizes existing local government resources, including funds for new career activities. Accomplishments of the project include:…

Social Science Research Institutes in the Quality American University. Final Technical Report.

ERIC Educational Resources Information Center

Totman, Theodore L.

The technical report presents a chapter outline and thesis summary of an investigation of social science research institutes in American universities. The bulk of the report presents the thesis in four sections. Section I proposes a typology of organized social research units (OSRUs) in the 11 universities studied. Dimensions used to classify the…

Expanding Horizons: A Program for Students Entering Nontraditional Technical Occupations. PY95 Final Detailed Report.

ERIC Educational Resources Information Center

Austin Community Coll., TX.

The Expanding Horizons Project at Austin Community College successfully achieved its goals for Project Year 1994-95. During the year, the project accomplished the following: raised public awareness of the need to overcome gender bias, promoted career opportunities in nontraditional technical occupations to more than 1,200 prospective students,…

Technical Assistance and Training from the Document Design Project. Final Report.

ERIC Educational Resources Information Center

American Institutes for Research, Washington, DC.

Contained in this report is a description of the technical assistance and training phase of the Document Design Project, a program funded by the National Institute of Education and intended to address and correct the readability problems posed by public documents. The first section of the report provides background material on the assistance and…

Determining and Validating Barriers to Post-Secondary Vocational, Technical and Adult Education Programs in Wisconsin. Final Report.

ERIC Educational Resources Information Center

Farning, Max; Borden, Sally

A consortium of five Wisconsin Vocational, Technical, and Adult Education (VTAE) Districts (Gateway, Indianhead, Mid-State, Milwaukee, and Southwest) were utilized to identify, verify, and alleviate barriers to enrollment. A VTAE survey in 1976 identified six major reasons for individuals' failing to attend school after indicating an interest in…

Sociometric Clique Identification. Final Report.

ERIC Educational Resources Information Center

Kadushin, Charles

This report consists of four parts. The first part is a non-technical summary of the basic problem and an attempted solution. The second part is a technical review of the literature and a description of the basic algorithm used in the solution. The third part describes the use of the Sociogram System. The fourth part describes the use of CHAIN, a…

An Assessment of Workplace Skills Acquired by Students of Vocational and Technical Education Institutions

ERIC Educational Resources Information Center

Bakar, Ab Rahim; Mohamed, Shamsiah; Hamzah, Ramlah

2013-01-01

This study was performed to identify the employability skills of technical students from the Industrial Training Institutes (ITI) and Indigenous People's Trust Council (MARA) Skills Training Institutes (IKM) in Malaysia. The study sample consisted of 850 final year trainees of IKM and ITI. The sample was chosen by a random sampling procedure from…

Wisconsin Technical College System Board Equity Staff Development Workshops and Services--Phase IV. Final Report.

ERIC Educational Resources Information Center

Baldus, Lorayne

A staff development program on gender equity was conducted for personnel in Wisconsin's technical colleges using the train-the-trainer method. The training took two approaches: a class for college personnel and career challenge training for project directors of single parent and displaced homemaker grants. The inservice class resulted in increased…

A Technical, User and Cost Comparison Study of Microfiche Duplicate Film Material. Final Report.

ERIC Educational Resources Information Center

Prevel, James J.

A technical, user and cost comparison study was undertaken to provide the Educational Resources Information Clearinghouse (ERIC) staff with data on silver halide, diazo, and vesicular type films for microfiche duplication. This information will allow ERIC to determine if diazo and/or vesicular films should be considered in producing ERIC duplicate…

Chemical Frustration. A Design Principle for the Discovery of New Complex Alloy and Intermetallic Phases, Final Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fredrickson, Daniel C

2015-06-23

Final technical report for "Chemical Frustration: A Design Principle for the Discovery of New Complex Alloy and Intermetallic Phases" funded by the Office of Science through the Materials Chemistry Program of the Office of Basic Energy Sciences.

Technical support for geopressured-geothermal activities in Louisiana. Final geological report, 1 November 1981-31 October 1982

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pilger, R.H.

1984-06-25

This report contains the component Site-Specific Studies-Geophysics, Diagenesis, Geochemistry. Work for Site-Specific Studies is finished, and the results presented in this report are final. Individual studies have been entered separately into the data base. (ACR)