DOCSLIB.ORG

Open Annebuboltz 11-24-08 Thesis.Pdf

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Open Annebuboltz 11-24-08 Thesis.Pdf The Pennsylvania State University The Graduate School Eberly College of Science MOLECULAR EVOLUTION OF VIRULENCE AND ANTIGENIC DIVERSITY IN BORDETELLA BRONCHISEPTICA A Dissertation in Biochemistry, Microbiology and Molecular Biology by Anne M. Buboltz © 2008 Anne M. Buboltz Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy December 2008 The dissertation of Anne M. Buboltz was reviewed and approved* by the following: Eric T. Harvill Associate Professor of Microbiology and Infectious Diseases Dissertation Adviser Chair of Committee Avery August Professor of Immunology Peter J. Hudson Professor of Biology Kenneth C. Keiler Associate Professor of Biochemistry and Molecular Biology Kathleen Postle Professor of Biochemistry and Molecular Biology Richard J. Frisque Professor of Molecular Virology and Head of the Department of Biochemistry, Microbiology and Molecular Biology * Signatures are on file in the Graduate School. ii ABSTRACT Bordetella is a genus of Betaproteobacteria consisting of nine species, many of which cause respiratory disease. The population structure of Bordetella is clonal, as the majority of strains fall into a small number of genotypic complexes. While strains of B. bronchiseptica are very closely related genetically, this species exhibits a high level of phenotypic diversity. For example, B. bronchiseptica has been isolated from the respiratory tract of more than 20 species of mammals, causes a wide-variety of severities of respiratory disease, strains can differ up to 100,000-fold in their lethal doses and can differ in their expression of virulence factors. Despite this phenotypic diversity exhibited by B. bronchiseptica strains, very few studies have mapped these traits to a phylogenetic tree to examine the evolution of these traits. Beyond that, even fewer studies have determined the underlying genetics factors associated with the phenotypic trait that was mapped which makes it impossible to determine if the phenotypic diversity is associated with the same genetic factor and to examine the molecular evolution of this trait. Therefore, using a wide range of approaches including a murine model of infection, comparative genomics and transcriptomics, and phylogenetics, the molecular evolution of the genetics factors that appear to contribute to phenotypic diversity, measured either in terms of virulence or antigenic variation, were examined. The first section shows that the gene encoding adenylate cylcase toxin (CyaA) was replaced with a novel operon via lateral gene transfer (LGT) in a hypovirulent lineage of B. bronchiseptica. The next section shows that the type three secretion system (TTSS) contributes to the increased virulence of a B. bronchiseptica lineage. Finally, we found that two distinct O-Antigen loci, which appear to encode antigenically distinct O-Antigen serotypes, which homologously recombine between B. bronchiseptica lineages. Combined, data contained herein suggest that lateral gene transfer, differential gene expression and homologous iii recombination all contribute to the phenotypic diversity of B. bronchiseptica strains. We discuss the implication of these findings on both within and outside the Bordetella field, consider the ecological forces that may cause and maintain this diversity and future avenues of research that result from these works. iv TABLE OF CONTENTS List of Figures ………………….……………………………………..………. ix List of Abbreviations…………………………………………………….…….. xi Acknowledgements……………………………………………………………. xiii Chapter 1. INTRODUCTION…………………………………………………. 1 Microbial Genetic Diversity…………………………………………… 2 The clonal paradigm of bacteria …………………………………. 2 Bacterial Recombination and its detection …………...………… 2 Methods to analyze genetic diversity in bacterial populations ... 4 Basic Phylogenetic Definitions ……………....………………... 5 Comparative Genomics of Pathogenic Bacteria……..…………. 6 What affects Microbial Diversity?………….……….…………. 6 Evolution of Virulence ……………………………..…………………. 7 What is virulence and how is it measured?………………….…. 7 What pressures affect virulence? ……………………..………… 8 Bringing Fields Together ……………..………………………………… 9 Bordetella …………………………….………………………………… 10 Disease and Vaccines ………………………………..…………. 10 Evolution and Population Structure of the classical bordetellae ... 11 Bordetella BvgAS and Virulence Factors ……………..………… 12 B. bronchiseptica phenotypic diversity ………………..………… 13 Preface ..………………………………………………..………… 13 References ……………………………………………………..………… 14 v Chapter 2. REPLACEMENT OF ADENYLATE CYCLASE TOXIN IN A LINEAGE OF BORDETELLA BRONCHISEPTICA.…………………………………………. 28 Abstract………………………………………………………………... 29 Introduction……………………………………………………………. 30 Materials and Methods………………………………………………… 33 Results…………………………………………………………………. 39 Discussion……………………………………………………………… 50 Authors and Contributions …..………………………………………… 55 References……………………………………………………………… 56 Chapter 3. TYPE III SECRETION SYSTEM CONTRIBUTES TO THE INCREASED VIRULENCE OF A BORDETELLA BRONCHISEPTICA LINEAGE.…......... 64 Abstract………………………………………………………………… 65 Introduction…………………………………………………………….. 66 Materials and Methods…………………………………………………. 69 Results………………………………………………………………….. 76 Discussion……………………………………………………………… 85 Author and Contributions ………………………………………………… 89 References……………………………………………………………… 90 Chapter 4. RECOMBINATION OF O-ANTIGEN LOCI IN BORDETELLA BRONCHISEPTICA AIDS IN EVASION OF CROSS-IMMUNITY.…………………………………. 98 Abstract……………………………………………………………….... 99 Introduction…………………………………………………………….. 100 Materials and Methods…………………………………………………. 103 vi Results………………………………………………………………….. 107 Discussion………………………………………………………………. 116 Authors and Contributions ………………………………………………. 120 References………………………………………………………………. 121 Chapter 5. SUMMARY AND SIGNIFICANCE………………………………... 127 Synopsis ………………………………….....……………….…………… 128 Implications ………………………………………………….…………... 129 Strains Representing B. bronchiseptica…………………………... 129 Clonal Diversity and Recombination References……..………….. 130 Vaccine Design ………………………………...……..………….. 131 Discussion Points …………………………………………….…………... 132 Evolutionary Ecology of Virulence …………....……..………….. 132 Evolutionary Ecology of Antigenic Variation ………..………….. 133 Future Directions ……………………………………………..………….. 134 Determining the Function of ptp genes ……..………..………….. 134 Determining other factors that increase ST32 strain virulence……. 135 Molecular Epidemiology of B. bronchiseptica ……..………….... 135 References ……………………………………………………………….. 138 Appendix ………………………………………………………………………… 145 Appendix A: Bordetella Strain List .………………………………….… 146 Appendix B: MLST primer list .………………………………………… 149 Appendix C: qRT-PCR, RT-PCR, PCR primer list for Chapter 2 ……… 150 Appendix D: Colonization of Nasal Cavity and Trachea by strains 253 vii and RB50 for Chapter 2………………………………….. 152 Appendix E: qRT-PCR Results for Chapter 2 ……………………….… 153 Appendix F: Western blot for protein expression of Ptx by Bordetella strains for Chapter 2 …………..………………………….. 154 Appendix G: Clinical Reports Accompanying ST32 strains 1289 and S308 for Chapter 3 ……………………………………….. 155 Appendix H: qRT-PCR Primer List for Chapter 3 …………………….. 157 Appendix I: qRT-PCR Results for Chapter 3 ………………………… 158 Appendix J: Colonization of Nasal Cavity and Trachea by strains 1289 and RB50 and Lungs by strains 448 and RB50 …………. 159 viii LIST OF FIGURES Figures: Chapter 2: Figure 2.1: LD50 and quantification of lung bacterial load of B. bronchiseptica strains RB50 and 253 ……………………………………………………………. 39 Figure 2.2: Whole Transcriptome Analysis and Adenylate Cyclase Toxin Expression in B. bronchiseptica strains RB50 and 253 ………………………………. 41 Figure 2.3: Comparative Genomic Hybridization and Adenylate Cyclase Toxin Locus in strains RB50 and 253 …………………………………………………. 43 Figure 2.4: Loss of cyaA in B. bronchiseptica strain lineage …………...…………… 45 Figure 2.5: Loss of cyaA among a lineage of B. bronchiseptica …………………….. 47 Figure 2.6: LD50 of B. bronchiseptica Sequence Type 27 strains and RB50 cyaA …. 48 Chapter 3: Figure 3.1: LD50 and quantification of lung bacterial load of B. bronchiseptica strains RB50 and 1289 ……………………………………………………………. 76 Figure 3.2: Whole Transcriptome Analysis and TTSS Expression in B. bronchiseptica strains RB50 and 1289 …………………………………………………… 78 Figure 3.3: TTSS mediated affect on cytotoxicity and virulence of B. bronchiseptica strains RB50 and 1289 ……………………………………………………. 80 Figure 3.4: MLST analysis of 61 Bordetella strains …………………………………. 82 Figure 3.5: TTSS mediated affect on virulence of B. bronchiseptica strain S308, a ST32 isolate ……………………………………………………………. 83 ix Chapter 4: Figure 4.1: Recognition of B. bronchiseptica strains RB50 and 1289 using antibodies from convalescent-phase serum by western blot and ELISA …………….. 107 Figure 4.2: Determination of O-Antigen serotype in 19 B. bronchiseptica strains using convalescent-phase serum by western blot ………………………………. 109 Figure 4.3: Comparative genomic hybridization and PCR analysis of LPS related- genes in B. bronchiseptica strains ……………………………………….. 110 Figure 4.4: Mapping the presence of the classical or alternative O-Antigen genes to ST in 49 B. bronchiseptica strains …………………………………………… 112 Figure 4.5: Effect of LPS vaccination and passive transfer of immune serum on numbers of B. bronchiseptica strains RB50, 1289 and RB50Δwbm ……………… 114 x LIST OF ABBREVIATIONS cDNA: complementary DNA CFU: Colony Forming Unit CGH: Comparative Genomic Hybridization CyaA: Adenylate Cyclase Toxin ELISA: Enzyme Linked Immunosorbent Assay g: gravity HRP: Horseradish Peroxidase IACUC: Institutional Animal
Load more

Recommended publications
  • Structural and Functional Effects of Bordetella Avium Infection in the Turkey Respiratory Tract William George Van Alstine Iowa State University
    Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 1987 Structural and functional effects of Bordetella avium infection in the turkey respiratory tract William George Van Alstine Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Animal Sciences Commons, and the Veterinary Medicine Commons Recommended Citation Van Alstine, William George, "Structural and functional effects of Bordetella avium infection in the turkey respiratory tract " (1987). Retrospective Theses and Dissertations. 11655. https://lib.dr.iastate.edu/rtd/11655 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. INFORMATION TO USERS While the most advanced technology has been used to photograph and reproduce this manuscript, the quality of the reproduction is heavily dependent upon the quality of the material submitted. For example: • Manuscript pages may have indistinct print. In such cases, the best available copy has been filmed. • Manuscripts may not always be complete. In such cases, a note will indicate that it is not possible to obtain missing pages. • Copyrighted material may have been removed from the manuscript. In such cases, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, and charts) are photographed by sectioning the original, beginning at the upper left-hand comer and continuing from left to right in equal sections with small overlaps.
    [Show full text]
  • Plant-Derived Benzoxazinoids Act As Antibiotics and Shape Bacterial Communities
    Supplemental Material for: Plant-derived benzoxazinoids act as antibiotics and shape bacterial communities Niklas Schandry, Katharina Jandrasits, Ruben Garrido-Oter, Claude Becker Contents Supplemental Tables 2 Supplemental Table 1. Phylogenetic signal lambda . .2 Supplemental Table 2. Syncom strains . .3 Supplemental Table 3. PERMANOVA . .6 Supplemental Table 4. PERMANOVA comparing only two treatments . .7 Supplemental Table 5. ANOVA: Observed taxa . .8 Supplemental Table 6. Observed diversity means and pairwise comparisons . .9 Supplemental Table 7. ANOVA: Shannon Diversity . 11 Supplemental Table 8. Shannon diversity means and pairwise comparisons . 12 Supplemental Table 9. Correlation between change in relative abundance and change in growth . 14 Figures 15 Supplemental Figure 1 . 15 Supplemental Figure 2 . 16 Supplemental Figure 3 . 17 Supplemental Figure 4 . 18 1 Supplemental Tables Supplemental Table 1. Phylogenetic signal lambda Class Order Family lambda p.value All - All All All All 0.763 0.0004 * * Gram Negative - Proteobacteria All All All 0.817 0.0017 * * Alpha All All 0 0.9998 Alpha Rhizobiales All 0 1.0000 Alpha Rhizobiales Phyllobacteriacae 0 1.0000 Alpha Rhizobiales Rhizobiacaea 0.275 0.8837 Beta All All 1.034 0.0036 * * Beta Burkholderiales All 0.147 0.6171 Beta Burkholderiales Comamonadaceae 0 1.0000 Gamma All All 1 0.0000 * * Gamma Xanthomonadales All 1 0.0001 * * Gram Positive - Actinobacteria Actinomycetia Actinomycetales All 0 1.0000 Actinomycetia Actinomycetales Intrasporangiaceae 0.98 0.2730 Actinomycetia Actinomycetales Microbacteriaceae 1.054 0.3751 Actinomycetia Actinomycetales Nocardioidaceae 0 1.0000 Actinomycetia All All 0 1.0000 Gram Positive - All All All All 0.421 0.0325 * Gram Positive - Firmicutes Bacilli All All 0 1.0000 2 Supplemental Table 2.
    [Show full text]
  • Bordetella Pertussis
    Hot et al. BMC Genomics 2011, 12:207 http://www.biomedcentral.com/1471-2164/12/207 RESEARCHARTICLE Open Access Detection of small RNAs in Bordetella pertussis and identification of a novel repeated genetic element David Hot1,2,3,4,5*, Stéphanie Slupek1,2,3,4,5, Bérénice Wulbrecht1,2,3,4,5, Anthony D’Hondt1,2,3,4,5, Christine Hubans5,6, Rudy Antoine1,2,3,4,5, Camille Locht1,2,3,4,5 and Yves Lemoine1,2,3,4,5 Abstract Background: Small bacterial RNAs (sRNAs) have been shown to participate in the regulation of gene expression and have been identified in numerous prokaryotic species. Some of them are involved in the regulation of virulence in pathogenic bacteria. So far, little is known about sRNAs in Bordetella, and only very few sRNAs have been identified in the genome of Bordetella pertussis, the causative agent of whooping cough. Results: An in silico approach was used to predict sRNAs genes in intergenic regions of the B. pertussis genome. The genome sequences of B. pertussis, Bordetella parapertussis, Bordetella bronchiseptica and Bordetella avium were compared using a Blast, and significant hits were analyzed using RNAz. Twenty-three candidate regions were obtained, including regions encoding the already documented 6S RNA, and the GCVT and FMN riboswitches. The existence of sRNAs was verified by Northern blot analyses, and transcripts were detected for 13 out of the 20 additional candidates. These new sRNAs were named Bordetella pertussis RNAs, bpr. The expression of 4 of them differed between the early, exponential and late growth phases, and one of them, bprJ2, was found to be under the control of BvgA/BvgS two-component regulatory system of Bordetella virulence.
    [Show full text]
  • Bordetella Plrsr Regulatory System Controls Bvgas Activity And
    Bordetella PlrSR regulatory system controls BvgAS PNAS PLUS activity and virulence in the lower respiratory tract M. Ashley Bonea,1, Aaron J. Wilkb,1,2, Andrew I. Peraulta, Sara A. Marlatta,3, Erich V. Schellera, Rebecca Anthouarda, Qing Chenc, Scott Stibitzc, Peggy A. Cottera,4, and Steven M. Juliob,4 aDepartment of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599; bDepartment of Biology, Westmont College, Santa Barbara, CA 93108; and cDivision of Bacterial, Parasitic, and Allergenic Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892 Edited by Scott J. Hultgren, Washington University School of Medicine, St. Louis, MO, and approved January 6, 2017 (received for review June 13, 2016) Bacterial pathogens coordinate virulence using two-component to collectively as vags) and lack of expression of BvgAS-repressed regulatory systems (TCS). The Bordetella virulence gene (BvgAS) genes (called vrgs), which includes those encoding flagella in – phosphorelay-type TCS controls expression of all known protein B. bronchiseptica. The Bvg phase occurs when the bacteria are “ virulence factor-encoding genes and is considered the master vir- grown at ≤26 °C or when millimolar concentrations of MgSO4 or ulence regulator” in Bordetella pertussis, the causal agent of pertus- nicotinic acid are added to the growth medium (referred to as – sis, and related organisms, including the broad host range pathogen “modulating conditions”). The Bvg phase is characterized by Bordetella bronchiseptica. We recently discovered an additional sen- expression of vrg loci and lack of expression of vags. The Bvg- sor kinase, PlrS [for persistence in the lower respiratory tract (LRT) intermediate (Bvgi) phase occurs at intermediate temperatures B.
    [Show full text]
  • Bordetella Pertussis (Pertussis)
    Bordetella pertussis (Pertussis) Heather L. Daniels, DO,* Camille Sabella, MD* *Center for Pediatric Infectious Diseases, Cleveland Clinic Children’s, Cleveland, OH Education Gaps 1. Clinicians must understand the changing epidemiology of pertussis and the reasons for the endemic and epidemic nature of infection despite widespread vaccination. 2. Clinicians must understand the strategies developed to prevent pertussis in those who are at high risk for complications. Objectives After completing this article, readers should be able to: 1. Recognize the antigenic components of pertussis. 2. Understand the changing epidemiology of the disease and the major factors contributing to this change. 3. Describe the clinical features during the natural progression of pertussis and the complications of infection. 4. List the options for laboratory testing of pertussis and their respective limitations. 5. List the recommended agents for antimicrobial treatment and postexposure chemoprophylaxis of pertussis. 6. Understand the rationale for the current pertussis vaccine recommendations. AUTHOR DISCLOSURE Drs Daniels and Sabella have disclosed no financial relationships relevant to this article. This commentary does not contain a discussion of an unapproved/investigative use of a INTRODUCTION commercial product/device. Bordetella pertussis is a fastidious gram-negative coccobacillus responsible for the ABBREVIATIONS respiratory infection commonly known as “whooping cough.” The organism is CDC Centers for Disease Control and spread by respiratory droplets and is highly contagious among close contacts. The Prevention typical incubation period is 7 to 10 days, but it may be as long as 21 days. Neither DTaP diphtheria, tetanus, and acellular natural infection nor pertussis vaccination results in long-lasting immunity, pertussis vaccine DTwP diphtheria, tetanus, and whole cell contributing to endemic infection and 3- to 5-year cycles of pertussis epidemics.
    [Show full text]
  • Stability, Structural and Functional Properties of a Monomeric, Calcium-Loaded Adenylate Cyclase Toxin, Cyaa, from Bordetella Pertussis
    Stability, structural and functional properties of a monomeric, calcium-loaded adenylate cyclase toxin, CyaA, from Bordetella pertussis. Sara E Cannella, Véronique Yvette Ntsogo Enguéné, Marilyne Davi, Christian Malosse, Ana Cristina Sotomayor Pérez, Julia Chamot-Rooke, Patrice Vachette, Dominique Durand, Daniel Ladant, Alexandre Chenal To cite this version: Sara E Cannella, Véronique Yvette Ntsogo Enguéné, Marilyne Davi, Christian Malosse, Ana Cristina Sotomayor Pérez, et al.. Stability, structural and functional properties of a monomeric, calcium- loaded adenylate cyclase toxin, CyaA, from Bordetella pertussis.. Scientific Reports, Nature Publishing Group, 2017, 7, pp.42065. 10.1038/srep42065. pasteur-01508525 HAL Id: pasteur-01508525 https://hal-pasteur.archives-ouvertes.fr/pasteur-01508525 Submitted on 14 Apr 2017 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution| 4.0 International License www.nature.com/scientificreports OPEN Stability, structural and functional properties of a monomeric, calcium–loaded
    [Show full text]
  • Product Sheet Info
    Product Information Sheet for NR-42462 Bordetella pertussis, Strain I036 Incubation: Temperature: 37°C Atmosphere: Aerobic with or without 5% CO2 Catalog No. NR-42462 Propagation: 1. Keep vial frozen until ready for use, then thaw. For research use only. Not for human use. 2. Transfer the entire thawed aliquot into a single tube of broth. Contributors: 3. Use several drops of the suspension to inoculate an Eric Harvill, Professor of Microbiology and Infectious agar slant and/or plate. 1 Disease, Department of Veterinary and Biomedical Sciences, 4. Incubate the tube (with shaking) , slant and/or plate at The Pennsylvania State University, University Park, 37°C for 2 to 7 days. Pennsylvania, USA Citation: Manufacturer: Acknowledgment for publications should read “The following BEI Resources reagent was obtained through BEI Resources, NIAID, NIH: Bordetella pertussis, Strain I036, NR-42462.” Product Description: Bacteria Classification: Alcaligenaceae, Bordetella Biosafety Level: 2 Species: Bordetella pertussis Appropriate safety procedures should always be used with Strain: I036 this material. Laboratory safety is discussed in the following Original Source: Bordetella pertussis (B. pertussis), strain publication: U.S. Department of Health and Human Services, I036 was isolated in 2012 from a nasopharyngeal swab of Public Health Service, Centers for Disease Control and a patient with whooping cough in Washington, USA.1 Prevention, and National Institutes of Health. Biosafety in Comments: The complete genome of B. pertussis, strain Microbiological and Biomedical Laboratories. 5th ed. I036 has been sequenced (GenBank: AXSH00000000).2 Washington, DC: U.S. Government Printing Office, 2009; see www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm. B.
    [Show full text]
  • Bordetella Bronchiseptica
    RESEARCH MODELS AND SERVICES Infectious Agent Sheet: Bordetella bronchiseptica Classification In guinea pigs, morbidity and mortality are most commonly Small Gram-negative rod seen in young guinea pigs, although clinical signs are rare in modern colonies even when Bordetella bronchiseptica Family is detected, suggesting that past disease outbreaks may Alcaligenaceae have been due to combined infections of Bordetella and some other agent. For example, most reports of Bordetella Affected species pneumonia in guinea pigs are from a time before guinea pig Clinically significant in guinea pigs and in rabbits. Of adenovirus pneumonia was recognized. Affected guinea minor importance as a natural infection in rats and mice. pigs appear ill, with ruffled fur, labored breathing, and Experimental reports only in gerbils. No reports in hamsters. anorexia. They may also have a mucopurulent or catarrhal exudate at the nares. At necropsy, cranioventral areas Frequency of the lungs are consolidated and there may be purulent Rare in modern laboratory animal facilities. More likely in exudate in the airways. The tympanic bullae may also be pet rodents and rabbits, especially those exposed to other affected. Histologic examination reveals a suppurative species (cats, dogs). Prevalence in wild rodent and rabbit bronchopneumonia with heterophilic and mononuclear populations is unknown. Possible transmission from human infiltration of the airways and affected alveoli. caretakers to animals. In rabbits, the pathogenicity of Bordetella is uncertain. It Transmission may contribute to “snuffles” (rabbit upper respiratory tract Transmission is via aerosol, direct contact, or contact with infections) and is often found as a co-infection with nasal secretions of infected animals. P.
    [Show full text]
  • Bordetella Progressively Worsened; the Patient Ultimately Required Endotracheal Intubation and Mechanical Ventilation
    DISPATCHES 10-day history of increasing shortness of breath and cough Isolation associated with mild hemoptysis. The initial chest radio- graph demonstrated an extensive left-sided infi ltrate, which of Bordetella progressively worsened; the patient ultimately required endotracheal intubation and mechanical ventilation. Bron- avium and Novel choscopy showed purulent secretions in the left mainstem bronchus, complete obstruction of the bronchus, and frothy Bordetella Strain secretions in the right airways. Gram-stained bronchoalve- from Patients olar lavage (BAL) fl uid showed many polymorphonuclear leukocytes but no organisms. Routine bacterial culture of with Respiratory the fl uid isolated a pure culture that was phenotypically identifi ed by API NFT rapid test strip (bioMérieux, Hazel- Disease wood, MO, USA) as B. avium. Antimicrobial drug–suscep- tibility testing performed using Etest (AB Biodisk, Solna, Amanda T. Harrington, Jaime A. Castellanos, Sweden) resulted in the following MICs: ceftriaxone 2 μg/ Tomasz M. Ziedalski, Jill E. Clarridge III, mL, azithromycin 4 μg/mL, piperacillin/tazobactam 0.125/4 and Brad T. Cookson μg/mL. The patient was initially treated with azithromycin Bordetella avium is thought to be strictly an avian and ceftriaxone and completed empiric therapy with pip- pathogen. However, 16S rRNA gene sequencing identi- eracillin/tazobactam and vancomycin. By day 7, the pa- fi ed 2 isolates from 2 humans with respiratory disease as tient’s respiratory status improved enough that he could be B. avium and a novel B. avium–like strain. Thus, B. avium extubated. Follow-up chest radiographs showed substantial and B. avium–like organisms are rare opportunistic human resolution of the left-sided infi ltrates, and on day 11, the pathogens.
    [Show full text]
  • Bordetella Bronchisepticaand
    SWINE TECHNICAL ARTICLES BORDETELLA BRONCHISEPTICA AND RESPIRATORY DISEASE IN SWINE Samantha J. Hau 1, Susan L. Brockmeier1 1 National Animal Disease Center, ARS, USDA, Ames, IA Corresponding Author ([email protected]) ABSTRACT Bordetella bronchiseptica is an ubiquitous pathogen of swine Bordetella bronchiseptica is an aerobic, Gram negative, rod- causing respiratory infections including rhinitis, tracheitis, shaped bacterium that is capable of causing respiratory bronchitis, and pneumonia. It spreads quickly between infections in a wide range of mammalian species (1). It is a animals, especially during commingling of naïve animals with member of the Betaproteobacteria class and closely related to subclinical carriers. B. bronchiseptica causes disease through the human pathogen Bordetella pertussis. B. bronchiseptica is the production of virulence factors, such as adhesins and an important cause of respiratory infections in swine, where it toxins, which is coordinated by the BvgAS system. In addition to can cause a range of diseases from asymptomatic colonization causing primary disease, B. bronchiseptica can exacerbate viral to severe pneumonia (2, 3). The prevalence of B. bronchiseptica respiratory infections and predispose animals to other bacterial in swine is high and it is globally distributed within the swine respiratory pathogens, which increases the clinical signifi cance industry (4-10). of B. bronchiseptica colonization. Disease is currently managed with antibiotic treatments and vaccination strategies; however, colonization is diff icult to clear and animals can remain colonized and shed B. bronchiseptica long term, acting as a source of infection for naïve animals within a herd. 5 DISEASES Bordetella bronchiseptica causes respiratory disease in swine cause more severe atrophy of the turbinates, which can result that can impact both the upper and lower respiratory tracts.
    [Show full text]
  • UK SMI ID 05: Identification of Bordetella Species
    UK Standards for Microbiology Investigations Identification of Bordetella species This publication was created by Public Health England (PHE) in partnership with the NHS. Identification | ID 5 | Issue no: 4 | Issue date: 10.08.2020 | Page: 1 of 18 © Crown copyright 2020 Identification of Bordetella species Acknowledgments UK Standards for Microbiology Investigations (UK SMIs) are developed under the auspices of 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. UK 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. PHE publications gateway number: GW-983 UK Standards for Microbiology Investigations are produced in association with: Identification | ID 5 | Issue no: 4 | Issue date: 10.08.2020 | Page: 2 of 18 Identification of Bordetella species Contents Acknowledgments ................................................................................................................
    [Show full text]
  • The Bvgs PAS Domain: an Independent Sensory Perception Module in the Bordetella Bvgas
    bioRxiv preprint doi: https://doi.org/10.1101/614891; this version posted April 23, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 The BvgS PAS domain: an independent sensory perception module in the Bordetella BvgAS 2 phosphorelay 3 4 M. Ashley Sobran1 and Peggy A. Cotter1,* 5 6 1 Department of Microbiology and Immunology, University of North Carolina Chapel Hill, Chapel 7 Hill, NC 27599 8 9 *Corresponding author: [email protected] 10 11 Keywords: Bordetella, two-component system, PAS domain, BvgAS, PlrSR, respiratory infection 12 Running Title: The BvgS PAS domain is an independent signal perception domain 13 14 15 16 17 18 19 20 21 22 bioRxiv preprint doi: https://doi.org/10.1101/614891; this version posted April 23, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 23 24 Abstract 25 26 To detect and respond to the diverse environments they encounter, bacteria often use two- 27 component regulatory systems (TCSs) to coordinate essential cellular processes required for 28 survival. In pathogenic Bordetella species, the BvgAS TCS regulates expression of hundreds of 29 genes, including those encoding all known protein virulence factors, and its kinase activity is 30 essential for respiratory infection. Maintenance of BvgS kinase activity in the lower respiratory 31 tract (LRT) depends on the function of another TCS, PlrSR. While the periplasmic venus fly-trap 32 domains of BvgS have been implicated in responding to so-called modulating signals in vitro 33 (nicotinic acid and MgSO4), a role for the cytoplasmic Per-Arnt-Sim (PAS) domain in signal 34 perception has not previously been demonstrated.
    [Show full text]