DOCSLIB.ORG

Characterization and Validation of Neisseria Gonorrhoeae Proteins

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Characterization and Validation of Neisseria Gonorrhoeae Proteins AN ABSTRACT OF THE DISSERTATION OF Igor H. Wierzbicki for the degree of Doctor of Philosophy in Pharmaceutical Sciences presented on September 28, 2016. Title: Characterization and Validation of Neisseria gonorrhoeae Proteins GmhAGC and NGO1985 as Molecular Targets for Development of Novel Anti-gonorrhea Therapeutic Interventions. Abstract approved: ______________________________________________________ Aleksandra E. Sikora The sexually transmitted disease gonorrhea, caused by the Gram-negative bacterium and obligate human pathogen Neisseria gonorrhoeae, remains a significant health and economic burden worldwide. In the absence of a protective vaccine, antimicrobial agents are the only pharmacological intervention for patients with gonorrhea. However, due to the remarkable ability of gonococcus to develop antibiotic resistance, infections caused by N. gonorrhoeae are believed to become untreatable in the near future. Identification and elucidation of the physiological function of novel N. gonorrhoeae proteins is critical for the formulation of new therapeutic interventions. This work focuses on characterization and validation of two gonococcal proteins, GmhAGC and NGO1985, as targets for development of new antibiotics and a vaccine antigen, respectively. The sedoheptulose-7-phosphate isomerase, GmhAGC, is the first enzyme in the biosynthesis of nucleotide-activated-glycero-manno-heptoses. We demonstrate that N. gonorrhoeae GmhAGC is essential for lipooligosaccharide (LOS) synthesis and pivotal for bacterial viability. Our crystallization studies have shown that GmhAGC forms a homo-tetramer in the closed conformation with four zinc ions in the active site. Site directed mutagenesis studies showed that active site residues E65 and H183 are important for LOS synthesis but not bacterial viability, suggesting that abolition of LOS synthesis is disconnected from the GmhAGC involvement in N. gonorrhoeae viability. NGO1985 was initially described as a hypothetical lipoprotein containing two BON (Bacterial OsmY and Nodulation) domains, hypothesized to be involved in maintaining bacterial cell envelope integrity. In our studies we demonstrate that NGO1985 is a surface exposed lipoprotein, conserved among diverse gonococcal isolates. Deletion of ngo1985 results in bacterial cell envelope defects leading to a pleiotropic phenotype including increased susceptibility to antimicrobial agents, decreased survival during in vitro growth conditions mimicking the human host environment, and high attenuation in the murine model of infection. NGO1985 interactome studies indicated a broad network of interactions including potential association with β-Barrel Assembly Machinery (Bam) complex, antibiotic efflux pump(s), and several lipoproteins. Furthermore, we demonstrate that NGO1985 does not undergo lipoprotein sorting according to the +2 residue of the lipobox motif as characterized for Escherichia coli. We determined that both BON domains, in their native orientation, are essential for NGO1985 functionality and stability. Finally, for the first time we have investigated the importance of the BON domains’ conserved glycine residues and showed that these amino acids play a critical role in protein stability. Based on the importance of both GmhAGC and NGO1985 on gonococcal physiology, we conclude that these proteins are promising molecular targets for development of new anti-gonorrhea interventions. ©Copyright by Igor H. Wierzbicki September 28, 2016 All Rights Reserved Characterization and Validation of Neisseria gonorrhoeae Proteins GmhAGC and NGO1985 as Molecular Targets for Development of Novel Anti-gonorrhea Therapeutic Interventions by Igor H. Wierzbicki A DISSERTATION submitted to Oregon State University in partial fulfillment of the requirements for the degree of Doctor of Philosophy Presented September 28, 2016 Commencement June 2017 Doctor of Philosophy dissertation of Igor H. Wierzbicki presented on September 28, 2016 APPROVED: Major Professor, representing Pharmaceutical Sciences Dean of the College of Pharmacy Dean of the Graduate School I understand that my dissertation will become part of the permanent collection of Oregon State University libraries. My signature below authorizes release of my dissertation to any reader upon request. Igor H. Wierzbicki, Author ACKNOWLEDGEMENTS I would like to thank Dr. Aleksandra Sikora for giving me the opportunity to do my Doctor of Philosophy education under her guidance. Thanks to her dedication in my training, as well as her never-ending patience with my actions and behavior, I was able to learn all of the aspects of being a successful scientist. All of my achievements, recognitions, and awards throughout the last four years would not be possible without her great input. I would also like to thank all members, past and present, of the Sikora laboratory for creating a welcoming atmosphere and always being there for me. I would like give special thanks to Rachael Ryner for being a wonderful undergraduate trainee. I enjoyed every moment of being her teacher and mentor. Watching her rapid progress made me more than proud. I want to thank everybody in the College of Pharmacy for helping me make it through my graduate education with special thanks to Debra Peters and Patricia Beaumont for brightening days with their smiles and help with all of the overwhelming paper work; Dr. McPhail for being part of my graduate committee, for the training, and help with all of my uncertainties related to the graduate program; and Dr. Adam Alani for being a wonderful and understanding mentor. I want to thank all of my friends that I have made here in Oregon for all of the hikes, camping trips, barbecues, potluck dinners, concerts, parties, movie nights, and all other spontaneous occasions. I am happy that I had the chance to meet and know you. I give thanks to my mother and grandparents for believing in me and supporting my life decisions. Lastly, but definitely not least, I want to thank my beloved wife, Dorota, for encouraging and convincing me to pursue my doctorate education. Without her patience, understanding, support, caring, and love I would not succeed. TABLE OF CONTENTS Page Chapter 1. General introduction…………………………………………………..…...1 Neisseria genus overview…....………………………………………………...2 Neisseria gonorrhoeae and gonorrhea...…………………………………........3 Treatment and prevention of gonorrhea………………………………………..6 Proteomic mining of cell envelopes and naturally released membrane vesicles for identification of novel targets for development of anti-gonorrhea interventions……………………………………………………….…10 NGO1985…...………………………………………………………………..12 β-barrel Assembly Machinery (BAM) complex……………………………...13 Nucleotide-activated-glycero-manno-heptoses and GmhA………………….15 Scope of dissertation…………………………………………………………16 Chapter 2. Functional and structural studies on the Neisseria gonorrhoeae GmhA, the first enzyme in the glycero-manno-heptose biosynthesis pathways, demonstrate a critical role in lipooligosacharide synthesis and gonococcal viability………20 Abstract...…………………………………………………………………….21 Introduction…………………………………………………………………..22 TABLE OF CONTENTS (Continued) Page Materials and Methods……………………………………………………….25 Results………………………………………………………………………..32 Discussion…………………………………………………………………....40 Acknowledgements…………………………………………………………..44 Chapter 3. NGO1985 is a surface-exposed lipoprotein interacting with BAM complex and a gonorrhea vaccine candidate…………………………………………...61 Abstract..……………………………………………………………………..62 Introduction…………………………………………………………………..63 Materials and Methods……………………………………………………….65 Results………………………………………………………………………..77 Discussion……………………………………………………………………85 Chapter 4. Dissecting the signal of lipoprotein sorting and function of BON domains in Neisseria gonorrhoeae lipoprotein NGO1985……………...…………….116 Abstract……………………………………………………………………..117 Introduction…………………………………………………………………118 TABLE OF CONTENTS (Continued) Page Materials and Methods……………………………………………………...120 Results………………………………………………………………………123 Discussion…………………………………………………………………..128 Chapter 5. Discussion and Conclusion………...……………………………………142 Overview……………………………………………………………………143 GmhAGC involvement in physiology of Neisseria species…………………..144 NGO1985 is a surface-exposed lipoprotein of N. gonorrhoeae…………..…147 NGO1985 lipobox +2 residue implication in protein localization and function……………………………………………………………..150 NGO1985 functions in maintaining cell envelope homeostasis…………….151 NGO1985 association with β-barrel Assembly Machinery (BAM) Complex…………………………………………………………….154 BON domains of NGO1985………………………………………………...156 Conclusions and future directions…………………………………….…….158 TABLE OF CONTENTS (Continued) Page Bibliography ……………………………………………….……………………….160 Appendices: Abstracts of Additional Manuscripts………………………………….189 LIST OF FIGURES Figure Page 1.1. Cell envelope of N. gonorrhoeae. ………...……..………………………………18 2.1. GmhAGC is pivotal for N. gonorrhoeae growth and LOS synthesis……………...45 2.2. Assessment of GmhAGC subcellular localization and expression patterns……….47 2.3. GmhAGC expression in a panel of N. gonorrhoeae isolates………………………49 2.4. Hindering GmhAGC isomerase activity does not influence N. gonorrhoeae growth………………………………………………………………………..50 2.5. Homologues of GmhA from N. gonorrhoeae and N. meningitidis function interchangeably………………………………………………………………51 2.6. The structure of N. gonorrhoeae GmhAGC……………………………………….52 S2.1. Purification of rGmhAGC……………………………………………………….56 S2.2. Loading controls for immunoblotting experiments…………………………….57 S2.3. Analysis of single nucleotide
Load more

Recommended publications
  • Official Nh Dhhs Health Alert
    THIS IS AN OFFICIAL NH DHHS HEALTH ALERT Distributed by the NH Health Alert Network [email protected] May 18, 2018, 1300 EDT (1:00 PM EDT) NH-HAN 20180518 Tickborne Diseases in New Hampshire Key Points and Recommendations: 1. Blacklegged ticks transmit at least five different infections in New Hampshire (NH): Lyme disease, Anaplasma, Babesia, Powassan virus, and Borrelia miyamotoi. 2. NH has one of the highest rates of Lyme disease in the nation, and 50-60% of blacklegged ticks sampled from across NH have been found to be infected with Borrelia burgdorferi, the bacterium that causes Lyme disease. 3. NH has experienced a significant increase in human cases of anaplasmosis, with cases more than doubling from 2016 to 2017. The reason for the increase is unknown at this time. 4. The number of new cases of babesiosis also increased in 2017; because Babesia can be transmitted through blood transfusions in addition to tick bites, providers should ask patients with suspected babesiosis whether they have donated blood or received a blood transfusion. 5. Powassan is a newer tickborne disease which has been identified in three NH residents during past seasons in 2013, 2016 and 2017. While uncommon, Powassan can cause a debilitating neurological illness, so providers should maintain an index of suspicion for patients presenting with an unexplained meningoencephalitis. 6. Borrelia miyamotoi infection usually presents with a nonspecific febrile illness similar to other tickborne diseases like anaplasmosis, and has recently been identified in one NH resident. Tests for Lyme disease do not reliably detect Borrelia miyamotoi, so providers should consider specific testing for Borrelia miyamotoi (see Attachment 1) and other pathogens if testing for Lyme disease is negative but a tickborne disease is still suspected.
    [Show full text]
  • Q Fever in Small Ruminants and Its Public Health Importance
    Journal of Dairy & Veterinary Sciences ISSN: 2573-2196 Review Article Dairy and Vet Sci J Volume 9 Issue 1 - January 2019 Copyright © All rights are reserved by Tolera Tagesu Tucho DOI: 10.19080/JDVS.2019.09.555752 Q Fever in Small Ruminants and its Public Health Importance Tolera Tagesu* School of Veterinary Medicine, Jimma University, Ethiopia Submission: December 01, 2018; Published: January 11, 2019 *Corresponding author: Tolera Tagesu Tucho, School of Veterinary Medicine, Jimma University, Jimma Oromia, Ethiopia Abstract Query fever is caused by Coxiella burnetii, it’s a worldwide zoonotic infectious disease where domestic small ruminants are the main reservoirs for human infections. Coxiella burnetii, is a Gram-negative obligate intracellular bacterium, adapted to thrive within the phagolysosome of the phagocyte. Humans become infected primarily by inhaling aerosols that are contaminated with C. burnetii. Ingestion (particularly drinking raw milk) and person-to-person transmission are minor routes. Animals shed the bacterium in urine and feces, and in very high concentrations in birth by-products. The bacterium persists in the environment in a resistant spore-like form which may become airborne and transported long distances by the wind. It is considered primarily as occupational disease of workers in close contact with farm animals or processing their be commenced immediately whenever Q fever is suspected. To prevent both the introduction and spread of Q fever infection, preventive measures shouldproducts, be however,implemented it may including occur also immunization in persons without with currently direct contact. available Doxycycline vaccines drugof domestic is the first small line ruminant of treatment animals for Q and fever.
    [Show full text]
  • Coxiella Burnetii
    SENTINEL LEVEL CLINICAL LABORATORY GUIDELINES FOR SUSPECTED AGENTS OF BIOTERRORISM AND EMERGING INFECTIOUS DISEASES Coxiella burnetii American Society for Microbiology (ASM) Revised March 2016 For latest revision, see web site below: https://www.asm.org/Articles/Policy/Laboratory-Response-Network-LRN-Sentinel-Level-C ASM Subject Matter Expert: David Welch, Ph.D. Medical Microbiology Consulting Dallas, TX [email protected] ASM Sentinel Laboratory Protocol Working Group APHL Advisory Committee Vickie Baselski, Ph.D. Barbara Robinson-Dunn, Ph.D. Patricia Blevins, MPH University of Tennessee at Department of Clinical San Antonio Metro Health Memphis Pathology District Laboratory Memphis, TN Beaumont Health System [email protected] [email protected] Royal Oak, MI BRobinson- Erin Bowles David Craft, Ph.D. [email protected] Wisconsin State Laboratory of Penn State Milton S. Hershey Hygiene Medical Center Michael A. Saubolle, Ph.D. [email protected] Hershey, PA Banner Health System [email protected] Phoenix, AZ Christopher Chadwick, MS [email protected] Association of Public Health Peter H. Gilligan, Ph.D. m Laboratories University of North Carolina [email protected] Hospitals/ Susan L. Shiflett Clinical Microbiology and Michigan Department of Mary DeMartino, BS, Immunology Labs Community Health MT(ASCP)SM Chapel Hill, NC Lansing, MI State Hygienic Laboratory at the [email protected] [email protected] University of Iowa [email protected] Larry Gray, Ph.D. Alice Weissfeld, Ph.D. TriHealth Laboratories and Microbiology Specialists Inc. Harvey Holmes, PhD University of Cincinnati College Houston, TX Centers for Disease Control and of Medicine [email protected] Prevention Cincinnati, OH om [email protected] [email protected] David Welch, Ph.D.
    [Show full text]
  • Ehrlichiosis and Anaplasmosis Are Tick-Borne Diseases Caused by Obligate Anaplasmosis: Intracellular Bacteria in the Genera Ehrlichia and Anaplasma
    Ehrlichiosis and Importance Ehrlichiosis and anaplasmosis are tick-borne diseases caused by obligate Anaplasmosis: intracellular bacteria in the genera Ehrlichia and Anaplasma. These organisms are widespread in nature; the reservoir hosts include numerous wild animals, as well as Zoonotic Species some domesticated species. For many years, Ehrlichia and Anaplasma species have been known to cause illness in pets and livestock. The consequences of exposure vary Canine Monocytic Ehrlichiosis, from asymptomatic infections to severe, potentially fatal illness. Some organisms Canine Hemorrhagic Fever, have also been recognized as human pathogens since the 1980s and 1990s. Tropical Canine Pancytopenia, Etiology Tracker Dog Disease, Ehrlichiosis and anaplasmosis are caused by members of the genera Ehrlichia Canine Tick Typhus, and Anaplasma, respectively. Both genera contain small, pleomorphic, Gram negative, Nairobi Bleeding Disorder, obligate intracellular organisms, and belong to the family Anaplasmataceae, order Canine Granulocytic Ehrlichiosis, Rickettsiales. They are classified as α-proteobacteria. A number of Ehrlichia and Canine Granulocytic Anaplasmosis, Anaplasma species affect animals. A limited number of these organisms have also Equine Granulocytic Ehrlichiosis, been identified in people. Equine Granulocytic Anaplasmosis, Recent changes in taxonomy can make the nomenclature of the Anaplasmataceae Tick-borne Fever, and their diseases somewhat confusing. At one time, ehrlichiosis was a group of Pasture Fever, diseases caused by organisms that mostly replicated in membrane-bound cytoplasmic Human Monocytic Ehrlichiosis, vacuoles of leukocytes, and belonged to the genus Ehrlichia, tribe Ehrlichieae and Human Granulocytic Anaplasmosis, family Rickettsiaceae. The names of the diseases were often based on the host Human Granulocytic Ehrlichiosis, species, together with type of leukocyte most often infected.
    [Show full text]
  • Avoidance of Mechanisms of Innate Immune Response by Neisseria Gonorrhoeae
    ADVANCEMENTS OF MICROBIOLOGY – POSTĘPY MIKROBIOLOGII 2019, 58, 4, 367–373 DOI: 10.21307/PM–2019.58.4.367 AVOIDANCE OF MECHANISMS OF INNATE IMMUNE RESPONSE BY NEISSERIA GONORRHOEAE Jagoda Płaczkiewicz* Department of Virology, Institute of Microbiology, Faculty of Biology, University of Warsaw Submitted in July, accepted in October 2019 Abstract: Neisseria gonorrhoeae (gonococcus) is a Gram-negative bacteria and an etiological agent of the sexually transmitted disease – gonorrhea. N. gonorrhoeae possesses many mechanism to evade the innate immune response of the human host. Most are related to serum resistance and avoidance of complement killing. However the clinical symptoms of gonorrhea are correlated with a significant pres- ence of neutrophils, whose response is also insufficient and modulated by gonococci. 1. Introduction. 2. Adherence ability. 3. Serum resistance and complement system. 4. Neutrophils. 4.1. Phagocytosis. 4.1.1. Oxygen- dependent intracellular killing. 4.1.2. Oxygen-independent intracellular killing. 4.2. Neutrophil extracellular traps. 4.3. Degranulation. 4.4. Apoptosis. 5. Summary UNIKANIE MECHANIZMÓW WRODZONEJ ODPOWIEDZI IMMUNOLOGICZNEJ PRZEZ NEISSERIA GONORRHOEAE Streszczenie: Neisseria gonorrhoeae (gonokok) to Gram-ujemna dwoinka będąca czynnikiem etiologicznym choroby przenoszonej drogą płciową – rzeżączki. N. gonorrhoeae posiada liczne mechanizmy umożliwiające jej unikanie wrodzonej odpowiedzi immunologicznej gospodarza. Większość z nich związana jest ze zdolnością gonokoków do manipulowania układem dopełniacza gospodarza oraz odpor- nością tej bakterii na surowicę. Jednakże symptomy infekcji N. gonorrhoeae wynikają między innymi z obecności licznych neutrofili, których aktywność jest modulowana przez gonokoki. 1. Wprowadzenie. 2. Zdolność adherencji. 3. Surowica i układ dopełniacza. 4. Neutrofile. 4.1. Fagocytoza. 4.1.1. Wewnątrzkomórkowe zabijanie zależne od tlenu. 4.1.2.
    [Show full text]
  • 3- Chlamydia, Syphilis & Gonorrhea (STD)
    3- Chlamydia, syphilis & gonorrhea (STD) Microbiology 435’s Teamwork Reproductive Block Learning Objectives: ● Know the causative agents of syphilis, gonorrhea and Chlamydia infections. ● Realize that these three infections are acquired through sexual intercourse. ● Know the pathogenesis of syphilis, gonorrhea and Chlamydia infection. ● Describe the clinical feature of the primary, secondary tertiary syphilis and complications. ● Recall the different diagnostic methods for the different stages of syphilis. ● Describe the clinical features of gonorrhea that affect only men, only women and those ones which affect both sexes. ● Describe the different laboratory tests for the diagnosis of gonorrhea ● Describe the morphology and the distinct life cycle of the Chlamydia. ● Realize what are the different genera, species and serotypes of the family Chlamydophila. ● Recognize that Chlamydia cause different diseases that affect the eye (causing trachoma) and the respiratory system (mainly cause a typical pneumonia). ● Know the different urogenital clinical syndromes caused by Chlamydia trachomatis that affect men, women and both sex. ● Realize that these urogenital syndromes are difficult to differentiate clinically from the similar ones caused by N.gonorrheae. ● Know the treatment of syphilis, gonorrhea and Chlamydia infections. ● Realize that there are no effective vaccines against all these three diseases. Important Resources: 435 females & males slides and ​ ​ ​ Males notes notes, wikipedia, ​ ​ Females notes Lippincott’s Illustrated Reviews: Microbiology- ​ ​ Extra Third Edition ​ ​ Editing file: Here ​ ​ Credit: Team members ​ Introduction (take-home message) ● Syphilis, Chlamydia and Gonorrhea are main STDs, caused by delicate organisms that cannot survive ​ ​ ​ outside the body. Infection may not be localized. ​ ​ ● Clinical presentation may be similar (urethral or genital discharge, ulcers). ● One or more organisms (Bacteria, virus, parasite) may be transmitted by sexual contact.
    [Show full text]
  • Molecular Characterization of the Burkholderia Cenocepacia Dcw Operon and Ftsz Interactors As New Targets for Novel Antimicrobial Design
    antibiotics Article Molecular Characterization of the Burkholderia cenocepacia dcw Operon and FtsZ Interactors as New Targets for Novel Antimicrobial Design Gabriele Trespidi y , Viola Camilla Scoffone y, Giulia Barbieri , Giovanna Riccardi, Edda De Rossi * and Silvia Buroni * Department of Biology and Biotechnology “Lazzaro Spallanzani”, University of Pavia, 27100 Pavia, Italy; [email protected] (G.T.); viola.scoff[email protected] (V.C.S.); [email protected] (G.B.); [email protected] (G.R.) * Correspondence: [email protected] (E.D.R.); [email protected] (S.B.) These authors contributed equally to this work. y Received: 20 October 2020; Accepted: 23 November 2020; Published: 24 November 2020 Abstract: The worldwide spread of antimicrobial resistance highlights the need of new druggable cellular targets. The increasing knowledge of bacterial cell division suggested the potentiality of this pathway as a pool of alternative drug targets, mainly based on the essentiality of these proteins, as well as on the divergence from their eukaryotic counterparts. People suffering from cystic fibrosis are particularly challenged by the lack of antibiotic alternatives. Among the opportunistic pathogens that colonize the lungs of these patients, Burkholderia cenocepacia is a well-known multi-drug resistant bacterium, particularly difficult to treat. Here we describe the organization of its division cell wall (dcw) cluster: we found that 15 genes of the dcw operon can be transcribed as a polycistronic mRNA from mraZ to ftsZ and that its transcription is under the control of a strong promoter regulated by MraZ. B. cenocepacia J2315 FtsZ was also shown to interact with the other components of the divisome machinery, with a few differences respect to other bacteria, such as the direct interaction with FtsQ.
    [Show full text]
  • Drug-Resistant Neisseria Gonorrhoeae Threat Level Urgent
    DRUG-RESISTANT NEISSERIA GONORRHOEAE THREAT LEVEL URGENT 550,000 1.14M $133.4M Estimated Total new Annual discounted drug-resistant infections lifetime direct infections each each year medical costs year Neisseria gonorrhoeae causes gonorrhea, a sexually transmitted disease (STD) that can result in life-threatening ectopic pregnancy and infertility, and can increase the risk of getting and giving HIV. WHAT YOU NEED TO KNOW EMERGING ANTIBIOTIC RESISTANCE ■ Gonorrhea has quickly developed resistance to all Gonorrhea rapidly develops resistance to antibiotics—ceftriaxone but one class of antibiotics, and half of all infections is the last recommended treatment. are resistant to at least one antibiotic. Tests to detect 35% 1980s: 2007: 2012: resistance are not available at time of treatment. Penicillin Ciprofloxacin Cefixime and tetracycline no longer no longer 30% no longer recommended recommended ■ Gonorrhea spreads easily. Some men and most women recommended as a first-line regimen do not have symptoms and may not know they are 25% infected, increasing spread. 20% ■ Untreated gonorrhea can cause serious and permanent health problems in women and men, including ectopic 15% pregnancy and infertility, and can spread to the blood resulting in cardiovascular and neurological problems. 10% Resistance or Emerging Resistance or Emerging Resistance Percent of Gonococcal Infections with Infections of Gonococcal Percent 5% 0% 2000 2002 2004 2006 2008 2010 2012 2014 2016 Azithromycin Ciprofloxacin Cefixime Penicillin Ceftriaxone Tetracycline STD Clinic
    [Show full text]
  • Purple Bacteria and Their Relatives”
    INTERNATIONALJOURNAL OF SYSTEMATICBACTERIOLOGY, July 1988, p. 321-325 Vol. 38, No. 3 0020-7713/88/03032 1-05$02.OOtO Copyright 0 1988, International Union of Microbiological Societies Proteobacteria classis nov. a Name for the Phylogenetic Taxon That Includes the “Purple Bacteria and Their Relatives” E. STACKEBRANDT,l R. G. E. MURRAY,2* AND H. G. TRUPER3 Lehrstuhl fur Allgemeine Mikrobiologie, Biologiezentrum, Christian-Albrechts Universitat, 2300 Kid, Federal Republic of Germany’; Department of Microbiology and Immunology, University of Western Ontario, London, Ontario, Canada N6A 5C12; and Institut fur Mikrobiulogie, Universitat Bonn, 5300 Bonn I, Federal Republic of Germany3 Proteobacteria classis nov. is suggested as the name for a new higher taxon to circumscribe the a, p, y, and 6 groups that are included among the phylogenetic relatives of the purple photosynthetic bacteria and as a suitable collective name for reference to that group. The group names (alpha, etc.) remain as vernacular terms at the level of subclass pending further studies and nomenclatural proposals. Phylogenetic interpretations derived from the study of the interim while the phylogenetic data are being integrated ribosomal ribonucleic acid (rRNA) sequences and oligonu- into formal bacterial taxonomy. It does not appear to be cleotide catalogs provide an important factual base for inappropriate or confusing to use the protean prefix because arrangements of higher taxa of bacteria (25, 26). A recent of the genus Proteus among the Proteobacteria; the reasons workshop organized by the International Committee on for use are clear enough. Systematic Bacteriology recognized that a particularly di- This new class is so far only definable in phylogenetic verse but related group of gram-negative bacteria, including terms.
    [Show full text]
  • Gonorrhea, Chlamydia, and Syphilis
    2019 GONORRHEA, CHLAMYDIA, AND SYPHILIS AND CHLAMYDIA, GONORRHEA, Dedication TAG would like to thank the National Coalition of STD Directors for funding and input on the report. THE PIPELINE REPORT Pipeline for Gonorrhea, Chlamydia, and Syphilis By Jeremiah Johnson Introduction The current toolbox for addressing gonorrhea, chlamydia, and syphilis is inadequate. At a time where all three epidemics are dramatically expanding in locations all around the globe, including record-breaking rates of new infections in the United States, stakeholders must make do with old tools, inadequate systems for addressing sexual health, and a sparse research pipeline of new treatment, prevention, and diagnostic options. Lack of investment in sexual health research has left the field with inadequate prevention options, and limited access to infrastructure for testing and treatment have allowed sexually transmitted infections (STIs) to flourish. The consequences of this underinvestment are large: according to the World Health Organization (WHO), in 2012 there were an estimated 357 million new infections (roughly 1 million per day) of the four curable STIs: gonorrhea, chlamydia, syphilis, and trichomoniasis.1 In the United States, the three reportable STIs that are the focus of this report—gonorrhea, chlamydia, and syphilis—are growing at record paces. In 2017, a total of 30,644 cases of primary and secondary (P&S) syphilis—the most infectious stages of the disease—were reported in the United States. Since reaching a historic low in 2000 and 2001, the rate of P&S syphilis has increased almost every year, increasing 10.5% during 2016–2017. Also in 2017, 555,608 cases of gonorrhea were reported to the U.S.
    [Show full text]
  • Pocket Guide to Clinical Microbiology
    4TH EDITION Pocket Guide to Clinical Microbiology Christopher D. Doern 4TH EDITION POCKET GUIDE TO Clinical Microbiology 4TH EDITION POCKET GUIDE TO Clinical Microbiology Christopher D. Doern, PhD, D(ABMM) Assistant Professor, Pathology Director of Clinical Microbiology Virginia Commonwealth University Health System Medical College of Virginia Campus Washington, DC Copyright © 2018 Amer i can Society for Microbiology. All rights re served. No part of this publi ca tion may be re pro duced or trans mit ted in whole or in part or re used in any form or by any means, elec tronic or me chan i cal, in clud ing pho to copy ing and re cord ing, or by any in for ma tion stor age and re trieval sys tem, with out per mis sion in writ ing from the pub lish er. Disclaimer: To the best of the pub lish er’s knowl edge, this pub li ca tion pro­ vi des in for ma tion con cern ing the sub ject mat ter cov ered that is ac cu rate as of the date of pub li ca tion. The pub lisher is not pro vid ing le gal, med i cal, or other pro fes sional ser vices. Any ref er ence herein to any spe cific com mer cial prod ucts, pro ce dures, or ser vices by trade name, trade mark, man u fac turer, or oth er wise does not con sti tute or im ply en dorse ment, rec om men da tion, or fa vored sta tus by the Ameri can Society for Microbiology (ASM).
    [Show full text]
  • Detection of Tick-Borne Pathogens of the Genera Rickettsia, Anaplasma and Francisella in Ixodes Ricinus Ticks in Pomerania (Poland)
    pathogens Article Detection of Tick-Borne Pathogens of the Genera Rickettsia, Anaplasma and Francisella in Ixodes ricinus Ticks in Pomerania (Poland) Lucyna Kirczuk 1 , Mariusz Piotrowski 2 and Anna Rymaszewska 2,* 1 Department of Hydrobiology, Faculty of Biology, Institute of Biology, University of Szczecin, Felczaka 3c Street, 71-412 Szczecin, Poland; [email protected] 2 Department of Genetics and Genomics, Faculty of Biology, Institute of Biology, University of Szczecin, Felczaka 3c Street, 71-412 Szczecin, Poland; [email protected] * Correspondence: [email protected] Abstract: Tick-borne pathogens are an important medical and veterinary issue worldwide. Environ- mental monitoring in relation to not only climate change but also globalization is currently essential. The present study aimed to detect tick-borne pathogens of the genera Anaplasma, Rickettsia and Francisella in Ixodes ricinus ticks collected from the natural environment, i.e., recreational areas and pastures used for livestock grazing. A total of 1619 specimens of I. ricinus were collected, including ticks of all life stages (adults, nymphs and larvae). The study was performed using the PCR technique. Diagnostic gene fragments msp2 for Anaplasma, gltA for Rickettsia and tul4 for Francisella were ampli- fied. No Francisella spp. DNA was detected in I. ricinus. DNA of A. phagocytophilum was detected in 0.54% of ticks and Rickettsia spp. in 3.69%. Nucleotide sequence analysis revealed that only one species of Rickettsia, R. helvetica, was present in the studied tick population. The present results are a Citation: Kirczuk, L.; Piotrowski, M.; part of a large-scale analysis aimed at monitoring the level of tick infestation in Northwest Poland.
    [Show full text]