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BD-CS-057, REV 0 | AUGUST 2017 | Page 1
EXPLIFY RESPIRATORY PATHOGENS BY NEXT GENERATION SEQUENCING Limitations Negative results do not rule out viral, bacterial, or fungal infections. Targeted, PCR-based tests are generally more sensitive and are preferred when specific pathogens are suspected, especially for DNA viruses (Adenovirus, CMV, HHV6, HSV, and VZV), mycobacteria, and fungi. The analytical sensitivity of this test depends on the cellularity of the sample and the concentration of all microbes present. Analytical sensitivity is assessed using Internal Controls that are added to each sample. Sequencing data for Internal Controls is quantified. Samples with Internal Control values below the validated minimum may have reduced analytical sensitivity or contain inhibitors and are reported as ‘Reduced Analytical Sensitivity’. Additional respiratory pathogens to those reported cannot be excluded in samples with ‘Reduced Analytical Sensitivity’. Due to the complexity of next generation sequencing methodologies, there may be a risk of false-positive results. Contamination with organisms from the upper respiratory tract during specimen collection can also occur. The detection of viral, bacterial, and fungal nucleic acid does not imply organisms causing invasive infection. Results from this test need to be interpreted in conjunction with the clinical history, results of other laboratory tests, epidemiologic information, and other available data. Confirmation of positive results by an alternate method may be indicated in select cases. Validated Organisms BACTERIA Achromobacter -
List of the Pathogens Intended to Be Controlled Under Section 18 B.E
(Unofficial Translation) NOTIFICATION OF THE MINISTRY OF PUBLIC HEALTH RE: LIST OF THE PATHOGENS INTENDED TO BE CONTROLLED UNDER SECTION 18 B.E. 2561 (2018) By virtue of the provision pursuant to Section 5 paragraph one, Section 6 (1) and Section 18 of Pathogens and Animal Toxins Act, B.E. 2558 (2015), the Minister of Public Health, with the advice of the Pathogens and Animal Toxins Committee, has therefore issued this notification as follows: Clause 1 This notification is called “Notification of the Ministry of Public Health Re: list of the pathogens intended to be controlled under Section 18, B.E. 2561 (2018).” Clause 2 This Notification shall come into force as from the following date of its publication in the Government Gazette. Clause 3 The Notification of Ministry of Public Health Re: list of the pathogens intended to be controlled under Section 18, B.E. 2560 (2017) shall be cancelled. Clause 4 Define the pathogens codes and such codes shall have the following sequences: (1) English alphabets that used for indicating the type of pathogens are as follows: B stands for Bacteria F stands for fungus V stands for Virus P stands for Parasites T stands for Biological substances that are not Prion R stands for Prion (2) Pathogen risk group (3) Number indicating the sequence of each type of pathogens Clause 5 Pathogens intended to be controlled under Section 18, shall proceed as follows: (1) In the case of being the pathogens that are utilized and subjected to other law, such law shall be complied. (2) Apart from (1), the law on pathogens and animal toxin shall be complied. -
Genetic and Functional Studies of the Mip Protein of Legionella
t1.ì. Genetic and Functional Studies of the Mip Protein of Legionella Rodney Mark Ratcliff, BSc (Hons)' MASM Infectious Diseases Laboratories Institute of Medical and Veterinary Science and Department of Microbiology and Immunology UniversitY of Adelaide. Adelaide, South Australia A thesis submitted to the University of Adelaide for the degree of I)octor of Philosophy 15'h March 2000 amended 14th June 2000 Colonies of several Legionella strains on charcoal yeast extract agar (CYE) after 4 days incubation at 37"C in air. Various magnifications show typical ground-glass opalescent appearance. Some pure strains exhibit pleomorphic growth or colour. The top two photographs demonstrate typical red (LH) and blue-white (RH) fluorescence exhibited by some species when illuminated by a Woods (IJV) Lamp. * t Table of Contents .1 Chapter One: Introduction .1 Background .............'. .2 Morphology and TaxonomY J Legionellosis ............. 5 Mode of transmission "..'....'. 7 Environmental habitat 8 Interactions between Legionella and phagocytic hosts 9 Attachment 11 Engulfment and internalisation.'.. 13 Intracellular processing 13 Intracellular replication of legionellae .. " "' " "' 15 Host cell death and bacterial release 18 Virulence (the Genetic factors involved with intracellular multiplication and host cell killing .20 icm/dot system) Legiolysin .25 Msp (Znn* metaloprotease) ...'..... .25 .28 Lipopolysaccharide .29 The association of flagella with disease.. .30 Type IV fimbriae.... .31 Major outer membrane proteins....'.......'. JJ Heat shock proteins'.'. .34 Macrophage infectivity potentiator (Mip) protein Virulenceiraits of Legionella species other than L. pneumophila..........' .39 phylogeny .41 Chapter One (continued): Introduction to bacterial classification and .41 Identificati on of Legionella...'.,..'.. .46 Phylogeny .52 Methods of phylogenetic analysis' .53 Parsimony methods.'.. .55 Distance methods UPGMA cluster analYsis.'.'... -
Aquascreen® Legionella Species Qpcr Detection Kit
AquaScreen® Legionella species qPCR Detection Kit INSTRUCTIONS FOR USE FOR USE IN RESEARCH AND QUALITY CONTROL Symbols Lot No. Cat. No. Expiry date Storage temperature Number of reactions Manufacturer INDICATION The AquaScreen® Legionella species qPCR Detection kit is specifically designed for the quantitative detection of several Legionella species in water samples prepared with the AquaScreen® FastExt- ract kit. Its design complies with the requirements of AFNOR T90-471 and ISO/TS 12869:2012. Legionella are ubiquitous bacteria in surface water and moist soil, where they parasitize protozoa. The optimal growth temperature lies between +15 and +45 °C, whereas these gram-negative bacteria are dormant below 20 °C and do not survive above 60 °C. Importantly, Legionella are well-known as opportunistic intracellular human pathogens causing Legionnaires’ disease and Pontiac fever. The transmission occurs through inhalation of contami- nated aerosols generated by an infected source (e.g. human-made water systems like shower- heads, sink faucets, heaters, cooling towers, and many more). In order to efficiently prevent Legionella outbreaks, water safety control measures need syste- matic application but also reliable validation by fast Legionella testing. TEST PRINCIPLE The AquaScreen® Legionella species Kit uses qPCR for quantitative detection of legionella in wa- ter samples. In contrast to more time-consuming culture-based methods, AquaScreen® assays need less than six hours including sample preparation and qPCR to reliably detect Legionella. Moreover, the AquaScreen® qPCR assay has proven excellent performance in terms of specificity and sensitivity: other bacterial genera remain undetected whereas linear quantification is obtai- ned up to 1 x 106 particles per sample, therefore requiring no material dilution. -
The Role of Lipids in Legionella-Host Interaction
International Journal of Molecular Sciences Review The Role of Lipids in Legionella-Host Interaction Bozena Kowalczyk, Elzbieta Chmiel and Marta Palusinska-Szysz * Department of Genetics and Microbiology, Institute of Biological Sciences, Faculty of Biology and Biotechnology, Maria Curie-Sklodowska University, Akademicka St. 19, 20-033 Lublin, Poland; [email protected] (B.K.); [email protected] (E.C.) * Correspondence: [email protected] Abstract: Legionella are Gram-stain-negative rods associated with water environments: either nat- ural or man-made systems. The inhalation of aerosols containing Legionella bacteria leads to the development of a severe pneumonia termed Legionnaires’ disease. To establish an infection, these bacteria adapt to growth in the hostile environment of the host through the unusual structures of macromolecules that build the cell surface. The outer membrane of the cell envelope is a lipid bilayer with an asymmetric composition mostly of phospholipids in the inner leaflet and lipopolysaccha- rides (LPS) in the outer leaflet. The major membrane-forming phospholipid of Legionella spp. is phosphatidylcholine (PC)—a typical eukaryotic glycerophospholipid. PC synthesis in Legionella cells occurs via two independent pathways: the N-methylation (Pmt) pathway and the Pcs pathway. The utilisation of exogenous choline by Legionella spp. leads to changes in the composition of lipids and proteins, which influences the physicochemical properties of the cell surface. This phenotypic plastic- ity of the Legionella cell envelope determines the mode of interaction with the macrophages, which results in a decrease in the production of proinflammatory cytokines and modulates the interaction with antimicrobial peptides and proteins. The surface-exposed O-chain of Legionella pneumophila sg1 LPS consisting of a homopolymer of 5-acetamidino-7-acetamido-8-O-acetyl-3,5,7,9-tetradeoxy-L- glycero-D-galacto-non-2-ulosonic acid is probably the first component in contact with the host cell that anchors the bacteria in the host membrane. -
Legionella Feeleii: Pneumonia Or Pontiac Fever? Bacterial Virulence Traits and Host Immune Response
Medical Microbiology and Immunology (2019) 208:25–32 https://doi.org/10.1007/s00430-018-0571-0 REVIEW Legionella feeleii: pneumonia or Pontiac fever? Bacterial virulence traits and host immune response Changle Wang1 · Xia Chuai1 · Mei Liang2 Received: 17 July 2018 / Accepted: 27 October 2018 / Published online: 1 November 2018 © Springer-Verlag GmbH Germany, part of Springer Nature 2018 Abstract Gram-negative bacterium Legionella is able to proliferate intracellularly in mammalian host cells and amoeba, which became known in 1976 since they caused a large outbreak of pneumonia. It had been reported that different strains of Legionella pneumophila, Legionella micdadei, Legionella longbeachae, and Legionella feeleii caused human respiratory diseases, which were known as Pontiac fever or Legionnaires’ disease. However, the differences of the virulence traits among the strains of the single species and the pathogenesis of the two diseases that were due to the bacterial virulence factors had not been well elucidated. L. feeleii is an important pathogenic organism in Legionellae, which attracted attention due to cause an outbreak of Pontiac fever in 1981 in Canada. In published researches, it has been found that L. feeleii serogroup 2 (ATCC 35849, LfLD) possess mono-polar flagellum, and L. feeleii serogroup 1 (ATCC 35072, WRLf) could secrete some exopolysaccharide (EPS) materials to the surrounding. Although the virulence of the L. feeleii strain was evidenced that could be promoted, the EPS might be dispensable for the bacteria that caused Pontiac fever. Based on the current knowledge, we focused on bacterial infection in human and murine host cells, intracellular growth, cytopathogenicity, stimulatory capacity of cytokines secre- tion, and pathogenic effects of the EPS ofL. -
Testing Different Membrane Filters for 16S Rrna Gene-Based Metabarcoding in Karstic Springs
Testing different membrane filters for 16S rRNA gene-based metabarcoding in karstic springs Oana Teodora Moldovan 1*, Andreea Baricz 2,3,4*, Edina Szekeres 2,3,4, Marius Kenesz 1, Marial Alexandra Hoaghia 5, Erika Andrea Levei 5, Ionuț Cornel Mirea 6, Ruxandra Năstase-Bucur 1, Traian Brad 1, Iulia Chiciudean 2,3, Horia Leonard Banciu 2,3 Figure S1. Piper diagram of the chemical elements for the studied springs Table S1. The relative abundance of the Bacteria phyla in the analyzed membranes, with the dominant Proteobacteria (see also Figure 3). BANPOTOC BAITA RAPOLTEL Phylum/Percentage AE20 AE21 AE22 AE23 AE24 AE25 AE26 AE4 AE27 BE11 BE12 BE18 BE16 BE15 BE13 BE19 BE17 BE14 CE29 CE31 CE32 CE30 CE34 CE33 CE2 CE28 CE35 CE3 non-identified 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 Acidobacteria 0,01 0,00 0,00 0,01 0,00 0,01 0,01 0,15 0,00 0,96 0,86 0,85 1,08 0,83 2,32 1,95 0,39 0,21 0,00 0,00 0,21 0,14 0,00 0,20 0,10 0,07 0,08 0,00 Actinobacteria 4,05 4,68 6,59 7,00 6,96 8,37 8,08 3,20 11,37 3,75 4,55 6,78 6,58 5,72 7,74 8,28 2,97 1,40 2,28 3,65 1,04 0,52 2,16 2,24 3,59 11,54 2,98 1,14 Aquificae 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 Armatimonadetes 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,01 0,01 0,00 0,00 0,00 0,01 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 0,00 Bacteroidetes 0,17 0,06 0,10 0,09 0,13 0,17 0,10 2,28 0,06 32,55 -
ESCMID Online Lecture Library © by Author ESCMID Online Lecture Library Latex Agglutination Test
The etiological agent: Legionella pneumophila and other Legionella spp. Valeria Gaia © by author National Reference Centre for Legionella ESCMIDc/o Online Microbiology Lecture laboratory Library Ente Ospedaliero Cantonale Bellinzona - Switzerland © by author ESCMID Online Lecture Library Hystory of Legionnaires’ Disease July 21st 1976 - Philadelphia • 58th Convention of the American Legion at the Bellevue-Stratford Hotel • > 4000 World War II Veterans with families & friends • 600 persons staying at the hotel © by author • ESCMIDJuly 23nd: convention Online closed Lecture Library • Several veterans showed symptoms of pneumonia Searching for the causative agent David Fraser: CDC – Atlanta •Influenza virus? •Nickel intoxication? •Toxin? o 2603 toxicology tests o 5120 microscopy exams o 990 serological tests© by author ESCMIDEverybody seems Online to agree: Lecture it’s NOT a bacterialLibrary disease! July 22nd – August 2nd •High fever •Coughing •Breathing difficulties •Chest pains •Exposed Population =© people by authorstaying in the lobby or outside the Bellevue Stratford Hotel «Broad Street Pneumonia» •221ESCMID persons were Online infected (182+39 Lecture «Broad StreetLibrary Pneumonia» ) 34 patients died (29+5) September 1976-January 1977 Joseph McDade: aims to rule out Q-fever (Rickettsiae) •Injection of “infected” pulmonary tissue in Guinea Pigs microscopy: Cocci and small Bacilli not significant at the time •Inoculation in embryonated eggs + antibiotics to inhibit the growth of contaminating bacteria No growth Microscopy on the -
Legionella and Non-Tuberculous Mycobacteria Using MALDI TOF MS (Matrix Assisted Laser Desorption Ionisation Time of Flight Mass Spectrometry)
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by OTHES DIPLOMARBEIT Titel der Diplomarbeit Establishment of a reference database for Acanthamoeba, Legionella and non-tuberculous mycobacteria using MALDI TOF MS (Matrix Assisted Laser Desorption Ionisation Time of Flight Mass Spectrometry) Verfasserin Dzenita HASANACEVIC angestrebter akademischer Grad Magistra der Naturwissenschaften (Mag.rer.nat.) Wien, 2012 Studienkennzahl lt. A 442 Studienblatt: Studienrichtung lt. Studienblatt: Anthropologie Betreuerin: Ass. Prof. Univ. Doz. Mag. Dr. Julia Walochnik Contents 1 ABBREVIATIONS ..................................................................................................... 5 2 INTRODUCTION ....................................................................................................... 6 2.1 Acanthamoeba .................................................................................................... 6 2.1.1 Classification ................................................................................................ 6 2.1.1.1 Phylogeny of Acanthamoeba ................................................................. 6 2.1.1.2 Methods of classification ....................................................................... 8 2.1.2 Ecology and geographical distribution ........................................................ 11 2.1.2.1 Life cycle ............................................................................................. 11 2.1.2.2 Trophozoites ...................................................................................... -
Methods Development for Unregulated Contaminants in Drinking Water
Method Development for Unregulated Contaminants in Drinking Water: Public Meeting and Webinar Held June 6, 2018 USEPA, Office of Ground Water and Drinking Water Office of Water (MLK 140) EPA 815-A-18-001 June 2018 Methods Development for Unregulated Contaminants in Drinking Water Methods Development for Unregulated Contaminants in Drinking Water Public Meeting and Webinar June 6, 2018 9:00 a.m. ‐ 3:00 p.m. ET U.S. EPA Office of Water and Office of Research and Development Welcome & SDWA Regulatory Process Brenda Parris, U.S. EPA Office of Ground Water and Drinking Water Technical Support Center Page 1 of 103 Methods Development for Unregulated Contaminants in Drinking Water Participating by Webinar • Listen‐only mode Figure 1 • Click on “+” next to “Questions” in the control panel (Figure 1) to submit questions/comments Figure 2 • Type a question in the box; click send (Figure 2) • Submit questions as soon as possible • Questions will be answered at the end of the presentations June 2018 U.S. Environmental Protection Agency Slide 3 of 206 Agenda 8:30‐9:00 Stakeholder Sign‐In Welcome & SDWA Regulatory Process Overview of Method Development EPA Method 542 EPA Methods 524.2/524.3/524.4 and 525.3 EPA Method 556.1 ~10:15‐10:30 Break EPA Method 540 & 543 EPA Methods 537 & 538 Method in Development: PFAS Method in Development 558: Ethyl carbamate (Urethane) and N‐Methyl‐2‐pyrrolidone Method in Development: Nonylphenols ~11:45‐12:45 Lunch Method in Development: Legionella Method in Development: Mycobacterium ~1:45‐2:00 Break 2:00‐3:00 Open Forum and Discussion Closing Remarks Page 2 of 103 Methods Development for Unregulated Contaminants in Drinking Water Overview • Regulatory background for UCMR • Safe Drinking Water Act (SDWA) authority • Relationships to: • Contaminant Candidate List (CCL) • Unregulated Contaminant Monitoring Rule (UCMR) • Regulatory Determination • Six‐Year Review June 2018 U.S. -
Ten New Species of Legionella DON J
INTERNATIONAL JOURNALOF SYSTEMATIC BACTERIOLOGY, Jan. 1985, p. 50-59 Vol. 35, No. 1 0020-7713/85/010050-10$02.00/0 Ten New Species of Legionella DON J. BRENNER,'" ARNOLD G. STEIGERWALT,l GEORGE W. GORMAN,' HAZEL W. WILKINSON,' WILLIAM F. BIBB,l MEREDETH HACKEL,2 RICHARD L. TYNDALL,3 JOYCE CAMPBELL,4 JAMES C. FEELEY,' W. LANIER THACKER,' PETER SKALIY,l WILLIAM T. MARTIN,' BONNIE J. BRAKE,' BARRY S. FIELDS,' HAROLD V. McEACHERN,~AND LINDA K. CORCORAN' Division of Bacterial Diseases, Center for Infectious Diseases, Centers for Disease Control, Atlanta, Georgia 30333'; Pathology Department, University Hospital, University of Michigan, Ann Arbor, Michigan 481 092; Environmental Sciences Division, Department of Zoology, University of Tennessee, Knoxville, Tennessee 379163; and Ofice of Public Health Laboratories and Epidemiology, Department of Social and Health Services, Seattle, Washington 981 044 Ten new Legionella species were characterized on the basis of biochemical reactions, antigens, cellular fatty acids, isoprenoid quinones, and deoxyribonucleic acid relatedness. Nine of the new species were isolated from the environment, and one, Legionella hackeliae, was isolated from a bronchial biopsy specimen obtained from a patient with pneumonia. The species all exhibited the following biochemical reactions typical of the legionellae: growth on buffered cysteine-yeast extract agar, but not on blood agar; growth requirement for cysteine; gram negative; nitrate negative; urease negative; nonfermentative; catalase positive; production of a brown pigment on tyrosine-containing yeast extract agar; liquefaction of gelatin; and motility. Legionella s4iritensis was weakly positive for hydrolysis of hippurate; the other species were hippurate negative. Legionella cherrii, Legionella steigerwaltii, and Legionella parisiensis exhibited bluish white autofluorescence. -
Pyrosequencing Demonstrated Complex Microbial Communities in a Membrane
M&E Papers in Press. Published online on March 18, 2011 doi:10.1264/jsme2.ME10205 1 2 3 Revised ME10205 4 5 6 Pyrosequencing Demonstrated Complex Microbial Communities in a Membrane 7 Filtration System for a Drinking Water Treatment Plant 8 1 1 1 1 9 SOONDONG KWON , EUNJEONG MOON , TAEK-SEUNG KIM , SEUNGKWAN HONG , and HEE- 1* 10 DEUNG PARK 11 12 1School of Civil, Environmental and Architectural Engineering,Proofs Korea University, Anam- 13 Dong, Seongbuk-Gu, Seoul 136-713, South Korea 14 15 (Received December 1, 2010 - Accepted ViewFebruary 17, 2011) 16 17 * Corresponding author. 18 E-mail: [email protected];Advance Tel: +82-2-3290-4861; Fax: +82-2-928-7656. 19 20 Running headline: Microbial Communities in a Filtration System 1 Copyright 2011 by the Japanese Society of Microbial Ecology / the Japanese Society of Soil Microbiology 1 Abstract 2 Microbial community composition in a pilot-scale microfiltration plant for drinking water 3 treatment was investigated using high-throughput pyrosequencing technology. Sequences of 4 16S rRNA gene fragments were recovered from raw water, membrane tank particulate matter, 5 and membrane biofilm, and used for taxonomic assignments, estimations of diversity, and the 6 identification of potential pathogens. Greater bacterial diversity was observed in each sample 7 (1,133 – 1,731 operational taxonomic units) than studies using conventional methods, 8 primarily due to the large number (8,164 – 22,275) of sequences available for analysis and 9 the identification of rare species. Betaproteobacteria predominated in the raw water (61.1%), 10 while Alphaproteobacteria were predominant in the membrane tank particulate matter 11 (42.4%) and membrane biofilm (32.8%).