DOCSLIB.ORG

Rapid Detection of Viable Legionella Pneumophila in Tap Water by a Qpcr and RT-PCR-Based Method

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Rapid Detection of Viable Legionella Pneumophila in Tap Water by a Qpcr and RT-PCR-Based Method Journal of Applied Microbiology ISSN 1364-5072 ORIGINAL ARTICLE Rapid detection of viable Legionella pneumophila in tap water by a qPCR and RT-PCR-based method R. Boss1 , A. Baumgartner1, S. Kroos2, M. Blattner2, R. Fretz2 and D. Moor1 1 Federal Food Safety and Veterinary Office, Berne, Switzerland 2 Food Safety and Veterinary Office, Canton of Basel-Landschaft, Liestal, Switzerland Keywords Abstract alternative method, Legionella pneumophila, qPCR, rapid, reverse transcription-PCR, rRNA, Aims: A molecular method for a rapid detection of viable Legionella viable, water. pneumophila of all serogroups in tap water samples was developed as an alternative to the reference method (ISO). Legionellae are responsible for Correspondence Legionnaires’ disease, a severe pneumonia in humans with high lethality. Renate Boss and Dominik Moor, Federal Food Methods and Results: The developed method is based on a nutritional Safety and Veterinary Office, Berne, stimulation and detection of an increase in precursor 16S rRNA as an Switzerland. E-mails: [email protected], and indicator for viability. For quantification, DNA was detected by qPCR. This [email protected] method was compared to the ISO method using water samples obtained from public sports facilities in Switzerland. The sensitivity and specificity were 91 2018/0489: received 6 March 2018, revised 4 and 97%, respectively, when testing samples for compliance with a May 2018 and accepted 23 May 2018 microbiological criterion of 1000 cell equivalents per l. Conclusion: The new method is sensitive and specific for Leg. pneumophila doi:10.1111/jam.13932 and allows results to be obtained within 8 h upon arrival, compared to one week or more by the ISO method. Significance and Impact of the Study: The method represents a useful tool for a rapid detection of viable Leg. pneumophila of all serogroups in water by molecular biology. It can be used as an alternative to the ISO method for official water analysis for legionellae and particularly when a short test time is required. Infection follows inhalation of contaminated water Introduction aerosols. Elderly or immunocompromised people, smok- Legionellosis is a bacterial disease with two clinical mani- ers and patients suffering from chronic respiratory ill- festations in humans: the self-limiting and nonpneumonic nesses are particularly at risk (Lanternier et al. 2017). The Pontiac fever and Legionnaires’ disease, characterized by best preventive method is to adjust temperatures in cold severe respiratory symptoms including pneumonia with water to below 20°C and to ensure a minimum water high lethality. The infectious agent is Legionella spp. and temperature of 60°C in boilers and preferably of 55°Cin predominantly Legionella pneumophila (WHO 2007; distribution pipes (WHO 2007; Bedard et al. 2015; Dominguez et al. 2009; Parr et al. 2015; ECDC 2016). Rhoads et al. 2015). To prevent scalding, temperature Legionellae naturally occur in environmental water regulator devices should be installed at points-of-use. sources and are well adapted to man-made water installa- Further aspects of prevention are appropriate construc- tions (WHO 2007). They grow in warm water of 25– tions with minimized water stagnation and materials that 45°C and are often found in buildings with convoluted do not support microbial growth (Bedard et al. 2015). water pipelines that are stagnant or rarely flushed parts Legionnaires’ disease occurs worldwide (WHO 2007) of the system (Rhoads et al. 2015). Predilection sites are with an assumed high number of unreported cases (Parr shower heads, but also pipes, taps, Jacuzzi tubs, and air et al. 2015). For the European Union and Norway, the conditioning installations (WHO 2007). incidence of infection was 1Á14 and 1Á35 per 105 © 2018 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology 1 This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. Detection of viable Leg. pneumophila R. Boss et al. inhabitants in 2013 and 2014 respectively (ECDC 2015, 2010). Upon stagnancy of growth, precursor rRNA syn- 2016). In Switzerland, where it is a reportable disease, it thesis stops but maturation goes on, draining the precur- showed a continuous increase to 5Á8 infections per 105 sor rRNA pool. Thus, the presence of precursor rRNA inhabitants in 2017, compared to an average of 0Á8(0Á3– can be used as a molecular indicator for physiological 1Á1) in the 1990s and 2Á1(0Á9–2Á8) between 2000 and activity and therefore viability of bacterial cells (Stroot 2009 respectively (FOPH 1990-2017, 2018). In Europe, and Oerther 2003; Lu et al. 2009; Cangelosi et al. 2010). approximately 70% of the reported infections are caused As water is a limited nutritional medium for legionellae, by the Leg. pneumophila serogroup (SG) 1, 20–30% by cell populations are barely in an exponential growth phase Leg. pneumophila SGs 2–16, and 5–10% by Legionella and the precursor rRNA pool at a low level or even drained spp., mainly Legionella micdadei (WHO 2007). Nonpneu- off (Al-Bana et al. 2014). The transfer of such starved bac- mophila Legionella spp. are predominantly involved in teria into a fresh nutritional medium will stimulate them, nosocomial infection (WHO 2007). resulting in a boosted rRNA synthesis. Dead cell is neither For preventive purposes, microbiological criteria for activated nor is their rRNA synthesis boosted. Such a stim- Legionella spp. in public accessible bath and shower water ulation step was included in the newly developed method. were recently stipulated in Switzerland (FSVO 2017). To It is followed by nucleic acid (NA) extraction and reverse test water samples for compliance with these microbio- transcription-PCR (RT-PCR) to detect the precursor 16S logical criteria, an international standard method is speci- rRNA. A shift between the cycle threshold (CT)ofan fied (ISO 11731) which detects and enumerates living unstimulated and a stimulated sample can be interpreted cells of the pathogen by culturing on selective agar plates as the presence of viable Leg. pneumophila cells. For quan- (ISO 2008). Alternative methods are allowed, when the tification, a real-time quantitative PCR (qPCR) was per- result’s evaluation does not influence the resulting deci- formed using the DNA fractions that were simultaneously sion. The ISO method is time consuming (up to 10 days) extracted from the same samples. and also has some other disadvantages (Kirschner 2016). The aim of the study was to develop and evaluate a rapid For example, legionellae can persist in a viable but not method to detect viable Leg. pneumophila in tap water sam- culturable (VBNC) status due to different reasons (Al- ples as an alternative to the bacteriological reference Bana et al. 2014; Li et al. 2015; Kirschner 2016), which method (ISO 11731). The two methods were compared by may result in false negative results. Under certain condi- analysing water samples from public sports facilities in a tions, VBNC legionellae are able to regain virulence and region of Switzerland with rather high incidence rates of infectiousness (Steinert et al. 1997; Ducret et al. 2014). legionellosis, that is, 7Á3 and 4Á6 infections per 105 inhabi- Molecular-based methods are faster but they usually tants in 2015 and 2016 respectively (FOPH 2018). detect both, viable and dead bacteria. The presence or number of dead cells might be of interest in some situa- Materials and methods tions but for the official control of water samples for legionellae, methods are needed which include only living Bacteriological detection of Leg. pneumophila cells. Methods using DNA intercalating reagents can dis- tinguish between living and dead bacterial cells but they Water samples were analyzed with the reference method have disadvantages. For example, the presence of biofilms ISO 11731-2:2008 (ISO 2008). In brief, 1 l of water was fil- can disturb detection (Taylor et al. 2014) and concentra- tered through a 0Á2 lm polycarbonate NucleporeTM mem- tion dependent cytotoxic effects of reagents are possible brane (Whatman Inc., Florham Park, NJ) and the bacterial (Yanez et al. 2011). Moreover, high cell counts, which cells then resuspended from the membrane. This suspension usually do not occur in field samples, are required for was plated on agar plates either directly or after acid or heat plausible analyses (Chang et al. 2009). pretreatment to minimize accompanying bacterial flora. Hence, as an alternative to the ISO method, a molecu- Plates were evaluated three times during the 10 days incu- lar detection method of viable Leg. pneumophila was bation period. One presumptive Legionella spp. per plate developed, based on PCR detection of a precursor 16S was confirmed by latex agglutination test (Oxoid, Pratteln, rRNA target that is specific for this species and occurs Switzerland and Microgen Bioproducts, Camberley, UK) only in living cells. Precursor rRNA represents a signifi- which identify the predominant Legionella spp. and allow cant fraction of the total microbial rRNA and is, because distinction between Leg. pneumophila SG 1 and SGs 2-15. of its higher stability, much easier to detect and handle than mRNA (Cangelosi et al. 2010). Precursor rRNA is Molecular detection of Leg. pneumophila synthesized by growing bacteria and its leader and tail sequences are subsequently removed during rRNA matu- A PCR system was designed that detects the precursor ration (Cangelosi and Brabant 1997; Cangelosi et al. region of the 16S rRNA sequence of Leg. pneumophila 2 © 2018 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology R. Boss et al. Detection of viable Leg. pneumophila which occurs threefold within its genome. Three copies Target concentration of the target were therefore regarded as one cell equiva- A 1 l water sample was filtered through a 0Á2 lm poly- lent (cellEq).
Load more

Recommended publications
  • Host-Adaptation in Legionellales Is 2.4 Ga, Coincident with Eukaryogenesis
    bioRxiv preprint doi: https://doi.org/10.1101/852004; this version posted February 27, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC 4.0 International license. 1 Host-adaptation in Legionellales is 2.4 Ga, 2 coincident with eukaryogenesis 3 4 5 Eric Hugoson1,2, Tea Ammunét1 †, and Lionel Guy1* 6 7 1 Department of Medical Biochemistry and Microbiology, Science for Life Laboratories, 8 Uppsala University, Box 582, 75123 Uppsala, Sweden 9 2 Department of Microbial Population Biology, Max Planck Institute for Evolutionary 10 Biology, D-24306 Plön, Germany 11 † current address: Medical Bioinformatics Centre, Turku Bioscience, University of Turku, 12 Tykistökatu 6A, 20520 Turku, Finland 13 * corresponding author 14 1 bioRxiv preprint doi: https://doi.org/10.1101/852004; this version posted February 27, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC 4.0 International license. 15 Abstract 16 Bacteria adapting to living in a host cell caused the most salient events in the evolution of 17 eukaryotes, namely the seminal fusion with an archaeon 1, and the emergence of both the 18 mitochondrion and the chloroplast 2. A bacterial clade that may hold the key to understanding 19 these events is the deep-branching gammaproteobacterial order Legionellales – containing 20 among others Coxiella and Legionella – of which all known members grow inside eukaryotic 21 cells 3.
    [Show full text]
  • Electronic Laboratory Reporting Use Case January 2011
    ILLINOIS HEALTH INFORMATION EXCHANGE Electronic Laboratory Reporting Use Case Electronic Laboratory Reporting and Health Information Exchange Illinois Health Information Exchange Public Health Work Group January 2011 Electronic Laboratory Reporting Use Case January 2011 Table of Contents 1.0 Executive Summary……………………………………………………………………….3 2.0 Introduction…………………………………………………………………...……………..5 3.0 Scope……………………………………..………………………………………………………5 4.0 Use Case Stakeholders…………………………………………………………….….....6 5.0 Issues and Obstacles……………………………………………………………………...8 6.0 Use Case Pre-Conditions .………………….…………………………………………...8 7.0 Use Case Post-Conditions.……………………………………………………………...9 8.0 Detailed Scenarios/Technical Specifications.………………………………10 9.0 Validation and Certification………………………………………………………...12 Appendix ………………………………………………………………………………………….....13 Page 2 Electronic Laboratory Reporting Use Case January 2011 1.0 Executive Summary This Use Case is a product of the Public Health Work Group (PHWG) of the Illinois Health Information Exchange (HIE) Advisory Committee. The Illinois HIE Advisory Committee was constituted as the diverse public healthcare stakeholder body providing input and recommendations on the creation of the Illinois Health Information Exchange Authority (“the Authority”) as the Illinois vehicle for designing and implementing electronic health information exchange in Illinois. The establishment of the Authority marks the formal transition of the work of the HIE Advisory Committee and the Work Groups into alignment with the provisions of Illinois
    [Show full text]
  • University of Nova Gorica Graduate School
    UNIVERSITY OF NOVA GORICA GRADUATE SCHOOL THE STUDY OF OPTIMAL TECHNOLOGICAL PROCEDURES OF INTERNAL PLUMBING SYSTEM DISINFECTION FACILITIES IN USE BY THE SENSITIVE HUMAN POPULATIONS MASTER´S THESIS Janez Škarja Mentor: Assis. Prof. Darko Drev Nova Gorica, 2016 UNIVERZA V NOVI GORICI FAKULTETA ZA PODIPLOMSKI ŠTUDIJ RAZISKAVA OPTIMALNIH TEHNOLOŠKIH POSTOPKOV DEZINFEKCIJE INTERNIH VODOVODNIH OMREŽIJ OBJEKTOV, KI JIH UPORABLJA OBČUTLJIVEJŠA POPULACIJA LJUDI MAGISTRSKO DELO Janez Škarja Mentor: doc.dr. Darko Drev Nova Gorica, 2016 SUMMARY In a developed world, water is used in a variety of installations and devices for the improvement of life standard. It is important for these elements to be suitably managed and maintained, otherwise they can present a risk to people's health. Although potable water from a public plumbing system coming via the water supply into an internal plumbing system normally is compliant with the regulations, the quality of water in an internal plumbing system often changes – water gets contaminated. There are several types of microorganisms that can grow in water. While most of them present no threat, there are some that can induce health issues in people. Internal plumbing systems with heated water and the possibility of aerosol releasing present a special concern in this matter. They carry a great potential for the growth and proliferation of bacteria from the genus Legionella that cause Legionnaires' disease and Pontiac Fever. Even the fact that Legionella infection has for several years been mentioned in the accident insurance conditions of insurance companies in relation to receiving the insurance fee as compensation for bed day, shows the extent and foremost the seriousness of the disease (5–15% mortality rate).
    [Show full text]
  • Isolation of Legionella Species from Drinking Water
    APPLIED AND ENVIRONMENTAL MICROBIOLOGY, OCt. 1984, p. 830-832 Vol. 48, No. 4 0099-2240/84/100830-03$02.00/0 Copyright © 1984, American Society for Microbiology Isolation of Legionella Species from Drinking Water S. C. HSU,* R. MARTIN, AND B. B. WENTWORTH Laboratory and Epidemiological Services Bureau, Lansing, Michigan 48909 Received 7 March 1984/Accepted 25 July 1984 Three different species of Legionella were recovered from samples of water taken from chlorinated public water supplies where no coliform bacteria were simultaneously detected. Five of 856 samples yielded Legionella isolates. Three isolates were identified as Legionella pneumophila serogroup 1, the fourth was identified as Legionella dumoffli, and the fifth was identified as Legionella jordanis. Studies to determine the survival of L. pneumophila Flint 1 serogroup 1 in tap water at various temperatures and in tap water with added sodium hypochlorite were done. These organisms were found to survive for 299 days in tap water at 24 and 5°C but not at 35°C. A concentration of at least 0.2 mg of residual chlorine per ml was required to eliminate at least 90% of L. pneumophila and Escherichia coli inocula in 2 h. Legionella species have been recognized as environmental gionella pneumophila serogroups 1 through 6, Legionella inhabitants, having been found not only in waters from bozemanii, Legionella micdadei, Legionella dumoffli, Le- cooling towers (5, 7, 8) and in plumbing systems of hospitals gionella gormanii, Legionellajordanis, and Legionella long- and hotels (3, 11) but also in natural water such as lakes, beachae serogroups 1 and 2. ponds, and streams (6, 10).
    [Show full text]
  • Front Matter
    JOURNAL OF CLINICAL MICROBIOLOGY VOLUME 21 * JUNE 1985 * NUMBER 6 Henry D. Isenberg, Editor in Chief (1989) Herman Friedman, Editor (1985) Peter B. Smith, Editor (1989) Long Island Jewish-Hillside College ofMedicine Centers for Disease Control Medical Center University ofSouth Florida Atlanta, Ga. New Hyde Park, N.Y. Tampa, Fla. Richard C. Tilton, Editor (1989) Steven D. Douglas, Editor (1988) Michael R. McGinnis, Editor (1985) University of Connecticut School of Children's Hospital ofPhiladelphia North Carolina Memorial Hospital Medicine Philadelphia, Pa. Chapel Hill, N.C. Farmington, Conn. Nathalie J. Schmidt, Editor (1985) California Department ofHealth, Berkeley, Calif. EDITORIAL BOARD Donald G. Ahearn (1987) Mario R. Escobar (1986) Victor Lorian (1987) J. Schachter (1986) Libero Ajelo (1985) Richard Facklam (1985) James D. MacLowry (1986) Joseph D. Schwartzman (1985) William L. Albritton (1987) J. J. Farmer (1986) Laurence R. McCarthy (1986) Alexis Shelokov (1985) Daniel Amsterdam (1986) Mary Jane Ferraro (1987) Kenneth McClatchy (1986) Maurice C. Shepard (1985) Ann M. Arvin (1987) Patricia Ferrieri (1986) Joseph E. McDade (1985) David M. Shlaes (1985) Lawrence Ash (1986) Sydney M. Finegold (1985) Jerry R. McGhee (1985) Salman Siddiqui (1987) Arthur L. Barry (1987) James Folds (1987) Joseph L. Melnick (1985) Marcelino F. Sierra (1987) Barry Beaty (1987) Marianne Forsgren (1987) Thomas Mitchell (1987) Robert M. Smibert I (1987) John E. Bennett (1985) Lynn S. Garcia (1986) Josephine A. Morello (1987) James W. Smith (1986) Merlin S. Bergdoll (1985) W. Lance George (1987) Stephen A. Morse (1986) Steven Specter (1986) Jennifer M. Best (1987) Gerald L. Gilardi (1986) C. Wayne Moss (1986) Leslie Spence (1985) E.
    [Show full text]
  • 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.
    [Show full text]
  • 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.
    [Show full text]
  • CGM-18-001 Perseus Report Update Bacterial Taxonomy Final Errata
    report Update of the bacterial taxonomy in the classification lists of COGEM July 2018 COGEM Report CGM 2018-04 Patrick L.J. RÜDELSHEIM & Pascale VAN ROOIJ PERSEUS BVBA Ordering information COGEM report No CGM 2018-04 E-mail: [email protected] Phone: +31-30-274 2777 Postal address: Netherlands Commission on Genetic Modification (COGEM), P.O. Box 578, 3720 AN Bilthoven, The Netherlands Internet Download as pdf-file: http://www.cogem.net → publications → research reports When ordering this report (free of charge), please mention title and number. Advisory Committee The authors gratefully acknowledge the members of the Advisory Committee for the valuable discussions and patience. Chair: Prof. dr. J.P.M. van Putten (Chair of the Medical Veterinary subcommittee of COGEM, Utrecht University) Members: Prof. dr. J.E. Degener (Member of the Medical Veterinary subcommittee of COGEM, University Medical Centre Groningen) Prof. dr. ir. J.D. van Elsas (Member of the Agriculture subcommittee of COGEM, University of Groningen) Dr. Lisette van der Knaap (COGEM-secretariat) Astrid Schulting (COGEM-secretariat) Disclaimer This report was commissioned by COGEM. The contents of this publication are the sole responsibility of the authors and may in no way be taken to represent the views of COGEM. Dit rapport is samengesteld in opdracht van de COGEM. De meningen die in het rapport worden weergegeven, zijn die van de auteurs en weerspiegelen niet noodzakelijkerwijs de mening van de COGEM. 2 | 24 Foreword COGEM advises the Dutch government on classifications of bacteria, and publishes listings of pathogenic and non-pathogenic bacteria that are updated regularly. These lists of bacteria originate from 2011, when COGEM petitioned a research project to evaluate the classifications of bacteria in the former GMO regulation and to supplement this list with bacteria that have been classified by other governmental organizations.
    [Show full text]
  • 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
    [Show full text]
  • ID 18 | Issue No: 3 | Issue Date: 14.04.15 | Page: 1 of 21 © Crown Copyright 2015 Identification of Legionella Species
    UK Standards for Microbiology Investigations Identification of Legionella species Issued by the Standards Unit, Microbiology Services, PHE Bacteriology – Identification | ID 18 | Issue no: 3 | Issue date: 14.04.15 | Page: 1 of 21 © Crown copyright 2015 Identification of Legionella species Acknowledgments UK Standards for Microbiology Investigations (SMIs) are developed under the auspices of Public Health England (PHE) working in partnership with the National Health Service (NHS), Public Health Wales and with the professional organisations whose logos are displayed below and listed on the website https://www.gov.uk/uk- standards-for-microbiology-investigations-smi-quality-and-consistency-in-clinical- laboratories. SMIs are developed, reviewed and revised by various working groups which are overseen by a steering committee (see https://www.gov.uk/government/groups/standards-for-microbiology-investigations- steering-committee). The contributions of many individuals in clinical, specialist and reference laboratories who have provided information and comments during the development of this document are acknowledged. We are grateful to the Medical Editors for editing the medical content. For further information please contact us at: Standards Unit Microbiology Services Public Health England 61 Colindale Avenue London NW9 5EQ E-mail: [email protected] Website: https://www.gov.uk/uk-standards-for-microbiology-investigations-smi-quality- and-consistency-in-clinical-laboratories PHE Publications gateway number: 2015013 UK Standards for Microbiology Investigations are produced in association with: Logos correct at time of publishing. Bacteriology – Identification | ID 18 | Issue no: 3 | Issue date: 14.04.15 | Page: 2 of 21 UK Standards for Microbiology Investigations | Issued by the Standards Unit, Public Health England Identification of Legionella species Contents ACKNOWLEDGMENTS .........................................................................................................
    [Show full text]
  • 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 ......................................................................................
    [Show full text]
  • Final Contaminant Candidate List 3 Microbes: Screening to PCCL
    Final Contaminant Candidate List 3 Microbes: Screening to the PCCL Office of Water (4607M) EPA 815-R-09-0005 August 2009 www.epa.gov/safewater EPA-OGWDW Final CCL 3 Microbes: EPA 815-R-09-0005 Screening to the PCCL August 2009 Contents Abbreviations and Acronyms ......................................................................................................... 2 1.0 Background and Scope ....................................................................................................... 3 2.0 Recommendations for Screening a Universe of Drinking Water Contaminants to Produce a PCCL.............................................................................................................................. 3 3.0 Definition of Screening Criteria and Rationale for Their Application............................... 5 3.1 Application of Screening Criteria to the Microbial CCL Universe ..........................................8 4.0 Additional Screening Criteria Considered.......................................................................... 9 4.1 Organism Covered by Existing Regulations.............................................................................9 4.1.1 Organisms Covered by Fecal Indicator Monitoring ..............................................................................9 4.1.2 Organisms Covered by Treatment Technique .....................................................................................10 5.0 Data Sources Used for Screening the Microbial CCL 3 Universe ................................... 11 6.0
    [Show full text]