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Legionella Shows a Diverse Secondary Metabolism Dependent on a Broad Spectrum Sfp-Type Phosphopantetheinyl Transferase
Legionella shows a diverse secondary metabolism dependent on a broad spectrum Sfp-type phosphopantetheinyl transferase Nicholas J. Tobias1, Tilman Ahrendt1, Ursula Schell2, Melissa Miltenberger1, Hubert Hilbi2,3 and Helge B. Bode1,4 1 Fachbereich Biowissenschaften, Merck Stiftungsprofessur fu¨r Molekulare Biotechnologie, Goethe Universita¨t, Frankfurt am Main, Germany 2 Max von Pettenkofer Institute, Ludwig-Maximilians-Universita¨tMu¨nchen, Munich, Germany 3 Institute of Medical Microbiology, University of Zu¨rich, Zu¨rich, Switzerland 4 Buchmann Institute for Molecular Life Sciences, Goethe Universita¨t, Frankfurt am Main, Germany ABSTRACT Several members of the genus Legionella cause Legionnaires’ disease, a potentially debilitating form of pneumonia. Studies frequently focus on the abundant number of virulence factors present in this genus. However, what is often overlooked is the role of secondary metabolites from Legionella. Following whole genome sequencing, we assembled and annotated the Legionella parisiensis DSM 19216 genome. Together with 14 other members of the Legionella, we performed comparative genomics and analysed the secondary metabolite potential of each strain. We found that Legionella contains a huge variety of biosynthetic gene clusters (BGCs) that are potentially making a significant number of novel natural products with undefined function. Surprisingly, only a single Sfp-like phosphopantetheinyl transferase is found in all Legionella strains analyzed that might be responsible for the activation of all carrier proteins in primary (fatty acid biosynthesis) and secondary metabolism (polyketide and non-ribosomal peptide synthesis). Using conserved active site motifs, we predict Submitted 29 June 2016 some novel compounds that are probably involved in cell-cell communication, Accepted 25 October 2016 Published 24 November 2016 differing to known communication systems. -
Microbial Diversity Under Extreme Euxinia: Mahoney Lake, Canada V
Geobiology (2012), 10, 223–235 DOI: 10.1111/j.1472-4669.2012.00317.x Microbial diversity under extreme euxinia: Mahoney Lake, Canada V. KLEPAC-CERAJ,1,2 C. A. HAYES,3 W. P. GILHOOLY,4 T. W. LYONS,5 R. KOLTER2 AND A. PEARSON3 1Department of Molecular Genetics, Forsyth Institute, Cambridge, MA, USA 2Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA, USA 3Department of Earth and Planetary Sciences, Harvard University, Cambridge, MA, USA 4Department of Earth and Planetary Sciences, Washington University, Saint Louis, MO, USA 5Department of Earth Sciences, University of California, Riverside, CA, USA ABSTRACT Mahoney Lake, British Columbia, Canada, is a stratified, 15-m deep saline lake with a euxinic (anoxic, sulfidic) hypolimnion. A dense plate of phototrophic purple sulfur bacteria is found at the chemocline, but to date the rest of the Mahoney Lake microbial ecosystem has been underexamined. In particular, the microbial community that resides in the aphotic hypolimnion and ⁄ or in the lake sediments is unknown, and it is unclear whether the sulfate reducers that supply sulfide for phototrophy live only within, or also below, the plate. Here we profiled distribu- tions of 16S rRNA genes using gene clone libraries and PhyloChip microarrays. Both approaches suggest that microbial diversity is greatest in the hypolimnion (8 m) and sediments. Diversity is lowest in the photosynthetic plate (7 m). Shallower depths (5 m, 7 m) are rich in Actinobacteria, Alphaproteobacteria, and Gammaproteo- bacteria, while deeper depths (8 m, sediments) are rich in Crenarchaeota, Natronoanaerobium, and Verrucomi- crobia. The heterogeneous distribution of Deltaproteobacteria and Epsilonproteobacteria between 7 and 8 m is consistent with metabolisms involving sulfur intermediates in the chemocline, but complete sulfate reduction in the hypolimnion. -
The Risk to Human Health from Free-Living Amoebae Interaction with Legionella in Drinking and Recycled Water Systems
THE RISK TO HUMAN HEALTH FROM FREE-LIVING AMOEBAE INTERACTION WITH LEGIONELLA IN DRINKING AND RECYCLED WATER SYSTEMS Dissertation submitted by JACQUELINE MARIE THOMAS BACHELOR OF SCIENCE (HONOURS) AND BACHELOR OF ARTS, UNSW In partial fulfillment of the requirements for the award of DOCTOR OF PHILOSOPHY in ENVIRONMENTAL ENGINEERING SCHOOL OF CIVIL AND ENVIRONMENTAL ENGINEERING FACULTY OF ENGINEERING MAY 2012 SUPERVISORS Professor Nicholas Ashbolt Office of Research and Development United States Environmental Protection Agency Cincinnati, Ohio USA and School of Civil and Environmental Engineering Faculty of Engineering The University of New South Wales Sydney, Australia Professor Richard Stuetz School of Civil and Environmental Engineering Faculty of Engineering The University of New South Wales Sydney, Australia Doctor Torsten Thomas School of Biotechnology and Biomolecular Sciences Faculty of Science The University of New South Wales Sydney, Australia ORIGINALITY STATEMENT '1 hereby declare that this submission is my own work and to the best of my knowledge it contains no materials previously published or written by another person, or substantial proportions of material which have been accepted for the award of any other degree or diploma at UNSW or any other educational institution, except where due acknowledgement is made in the thesis. Any contribution made to the research by others, with whom 1 have worked at UNSW or elsewhere, is explicitly acknowledged in the thesis. I also declare that the intellectual content of this thesis is the product of my own work, except to the extent that assistance from others in the project's design and conception or in style, presentation and linguistic expression is acknowledged.' Signed ~ ............................ -
Susceptibility and Resistance Data
toku-e logo For a complete list of references, please visit antibiotics.toku-e.com Levofloxacin Microorganism Genus, Species, and Strain (if shown) Concentration Range (μg/ml)Susceptibility and Aeromonas spp. 0.0625 Minimum Inhibitory Alcaligenes faecalis 0.39 - 25 Bacillus circulans Concentration0.25 - 8 (MIC) Data Bacillus subtilis (ATCC 6051) 6.25 Issue date 01/06/2020 Bacteroides capillosus ≤0.06 - >8 Bacteroides distasonis 0.5 - 128 Bacteroides eggerthii 4 Bacteroides fragilis 0.5 - 128 Bacteroides merdae 0.25 - >32 Bacteroides ovatus 0.25 - 256 Bacteroides thetaiotaomicron 1 - 256 Bacteroides uniformis 4 - 128 Bacteroides ureolyticus ≤0.06 - >8 Bacteroides vulgatus 1 - 256 Bifidobacterium adolescentis 0.25 - >32 Bifidobacterium bifidum 8 Bifidobacterium breve 0.25 - 8 Bifidobacterium longum 0.25 - 8 Bifidobacterium pseudolongum 8 Bifidobacterium sp. 0.25 - >32 Bilophila wadsworthia 0.25 - 16 Brevibacterium spp. 0.12 - 8 Brucella melitensis 0.5 Burkholderia cepacia 0.25 - 512 Campylobacter coli 0.015 - 128 Campylobacter concisus ≤0.06 - >8 Campylobacter gracilis ≤0.06 - >8 Campylobacter jejuni 0.015 - 128 Campylobacter mucosalis ≤0.06 - >8 Campylobacter rectus ≤0.06 - >8 Campylobacter showae ≤0.06 - >8 Campylobacter spp. 0.25 Campylobacter sputorum ≤0.06 - >8 Capnocytophaga ochracea ≤0.06 - >8 Capnocytophaga spp. 0.006 - 2 Chlamydia pneumonia 0.125 - 1 Chlamydia psittaci 0.5 Chlamydia trachomatis 0.12 - 1 Chlamydophila pneumonia 0.5 Citrobacter diversus 0.015 - 0.125 Citrobacter freundii ≤0.00625 - >64 Citrobacter koseri 0.015 - -
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. -
Immunoproteomic Identification of Biomarkers for Diagnosis of Legionellosis
Immunoproteomic identification of biomarkers for diagnosis of legionellosis Submitted in total fulfilment of the requirements for the degree of Doctor of philosophy by Kaylass Poorun Department of Chemistry and Biotechnology Faculty of Science, Engineering and Technology Swinburne University of Technology Australia 2014 Abstract Abstract Legionellosis, a disease with significant mortality and morbidity rates, is considered to be the second most frequent cause of severe community-acquired pneumonia. It is difficult to distinguish from other types of pneumonia due to similar clinical manifestations. Several studies have demonstrated the inadequacies of current diagnostic tests for confirming Legionella infections. This study was aimed at identifying biomarkers that can be used in an improved test. A comparative proteomic analysis, using DIGE, was carried out between L. pneumophila ATCC33152 and L. longbeachae NSW150 and D4968 isolates. While many homologous proteins were found to be commonly expressed, numerous others were identified to be differentially expressed under similar in vitro conditions suggesting that the two species have different lifestyles and infection strategies. The bacterial immunoglobulin domain containing protein, found to share sequence homology to Type V secretion proteins intimin and invasin, is not known to be present in Legionella. Human sera containing antibodies against Legionella from a set of blind samples were identified by ELISA. Downstream analyses revealed that diverse immunogens may be responsible for eliciting immune response in different Legionella species which in turn show little to no congeneric cross-reactivity. To the best of our knowledge, this is a unique finding not previously reported. Several serological diagnostic tests currently in use do not include many Legionella species in their testing panel, which may be a reason for many Legionella species being under-reported. -
Studies on Microbial Prospecting for Exobiopolymeric Flocculants
Studies on microbial prospecting for exobiopolymeric flocculants A thesis submitted in fulfillment of the requirement for the award of the degree of DOCTOR OF PHILOSOPHY IN BIOTECHNOLOGY Santosh Pathak (Regn No. 9051005) Department of Biotechnology and Environmental Sciences, Thapar University, Patiala –147004 Punjab (India) 2011 Acknowledgement I pay my reverence and gratitude to the Almighty for his everlasting blessings. This thesis arose out of years of research that has been done since I joined Thapar University. I have known a great number of people during these many years, whose contribution in assorted ways to the research and the making of the thesis deserved special mention. It is a pleasure to convey my gratitude to them all in my humble acknowledgment. Firstly, I offer my gratitude to my esteemed supervisor Dr. Moushumi Ghosh, Associate Professor, Thapar University, Patiala, for her supervision, advice, and guidance from the very early stage of this research as well as giving me extraordinary experiences throughout the work. Above all and the most needed, she provided me unflinching encouragement and support in various ways. Her truly scientific intuition has made her as a constant oasis of ideas and passions in Science, which exceptionally inspire and enrich my growth as a student, a researcher and a scientist want to be. I am indebted to her insightful comments, and for putting so much energy for this work. I take this opportunity to thank Dr. Abhijit Mukherjee, Director, Thapar University, for all the facilities which have been immensely helpful for the completion of my work. I express my gratitude to Dr. -
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. -
Biodiversidad Bacteriana Marina: Nuevos Taxones Cultivables
Departamento de Microbiología y Ecología Colección Española de Cultivos Tipo Doctorado en Biotecnología Biodiversidad bacteriana marina: nuevos taxones cultivables Directores de Tesis David Ruiz Arahal Mª Jesús Pujalte Domarco Mª Carmen Macián Rovira Teresa Lucena Reyes Tesis Doctoral Valencia, 2012 Dr. David Ruiz Arahal , Profesor Titular del Departamento de Microbiología y Ecología de la Universidad de Valencia, Dra. María Jesús Pujalte Domarco , Catedrática del Departamento de Microbiología y Ecología de la Universidad de Valencia, y Dra. Mª Carmen Macián Rovira , Técnico Superior de Investigación de la Colección Española de Cultivos Tipo de la Universidad de Valencia, CERTIFICAN: Que Teresa Lucena Reyes, Licenciada en Ciencias Biológicas por la Universidad de Valencia, ha realizado bajo su dirección el trabajo titulado “Biodiversidad bacteriana marina: nuevos taxones cultivables”, que presenta para optar al grado de Doctor en Ciencias Biológicas por la Universidad de Valencia. Y para que conste, en el cumplimiento de la legislación vigente, firman el presente certificado en Valencia, a. David Ruiz Arahal Mª Jesús Pujalte Domarco Mª Carmen Macián Rovira Relación de publicaciones derivadas de la presente Tesis Doctoral Lucena T , Pascual J, Garay E, Arahal DR, Macián MC, Pujalte MJ (2010) . Haliea mediterranea sp. nov., a marine gammaproteobacterium. Int J Syst Evol Microbiol 60 , 1844-8. Lucena T, Pascual J, Giordano A, Gambacorta A, Garay E, Arahal DR, Macián MC, Pujalte MJ (2010) . Euzebyella saccharophila gen. nov., sp. nov., a marine bacterium of the family Flavobacteriaceae . Int J Syst Evol Microbiol 60 , 2871-6. Lucena T, Ruvira MA, Pascual J, Garay E, Macián MC, Arahal DR, Pujalte MJ (2011) . Photobacterium aphoticum sp. -
The Flagellar Regulon of Legionella—A Review
REVIEW published: 20 October 2017 doi: 10.3389/fcimb.2017.00454 The Flagellar Regulon of Legionella—A Review Sandra Appelt 1 and Klaus Heuner 2* 1 Highly Pathogenic Microorganisms, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Berlin, Germany, 2 Cellular Interactions of Bacterial Pathogens, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Berlin, Germany The Legionella genus comprises more than 60 species. In particular, Legionella pneumophila is known to cause severe illnesses in humans. Legionellaceae are ubiquitous inhabitants of aquatic environments. Some Legionellaceae are motile and their motility is important to move around in habitats. Motility can be considered as a potential virulence factor as already shown for various human pathogens. The genes of the flagellar system, regulator and structural genes, are structured in hierarchical levels described as the flagellar regulon. Their expression is modulated by various environmental factors. For L. pneumophila it was shown that the expression of genes of the flagellar regulon is modulated by the actual growth phase and temperature. Especially, flagellated Legionella are known to express genes during the transmissive phase of growth that are involved in the expression of virulence traits. It has been demonstrated that the alternative sigma-28 factor is part of the link between virulence expression and motility. In the following review, the structure of the flagellar regulon of L. pneumophila is discussed and compared to other flagellar systems of different Legionella species. Recently, it has been described that Legionella micdadei and Legionella fallonii contain a second putative partial flagellar Edited by: system. Hence, the report will focus on flagellated and non-flagellated Legionella strains, Matthias P.